Map

Cocaine, a commonly abused psychostimulant, interacts with not only transporters for dopamine, serotonin and norepinephrine but also several receptors and channels. However, the molecular mechanisms underlying the various eVects of cocaine remain to be clariWed. Using the Xenopus oocyte expression assay, we investigated the eVects of cocaine on G protein-activated inwardly rectifying K + (GIRK) channels, which regulate neuronal excitability and the heart rate. In oocytes injected with mRNAs for GIRK1/GIRK2, GIRK2 or GIRK1/GIRK4 subunits, cocaine reversibly reduced basal GIRK inward currents. The inhibition by cocaine at the toxic levels was concentration-dependent, but voltage-independent and time-independent during each voltage pulse. However, methylphenidate, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) at their toxic concentrations had little eVect on the channels. Additionally, Kir1.1 and Kir2.1 channels were insensitive to all of the drugs. The inhibition by cocaine, which exists mainly in a protonated form at pH 7.4, was not aVected by extracellular pH 9, at which the proportion of the uncharged form increases, suggesting the inhibition by both forms with similar eVectiveness, and at physiological pH the eVect being predominantly due to the protonated cocaine. Our results suggest that inhibition of GIRK channels by cocaine may contribute to some of its toxic eVects.

This paper develops a comprehensive and objective picture of bird distributions relative to habitats across Britain. Bird species presence/absence data from an extensive field survey and habitat data from the remotely sensed UK Land Cover Map 2000 were analysed in 36,920 tetrads (2 km  2 km) across Britain (a 65% sample of Britain's c. 240 000 km 2 ). Cluster analysis linked birds to generalised landscapes based on distinctive habitat assemblages. Maps of the clusters showed strong regional patterns associated with the habitat assemblages. Cluster centroid coordinates for each bird species and each habitat were combined across clusters to derive individualised bird-habitat preference indices and examine the importance of individual habitats for each bird species. Even rare species and scarce habitats showed successful linkages. Results were assessed against published accounts of bird-habitat relations. Objective corroboration strongly supported the associations. Relatively scarce coastal and wetland habitats proved particularly important for many birds. However, extensive arable farmland and woodland habitats were also favoured by many species, despite reported declines in bird numbers in these habitats. The fact that habitat-specialists do not or cannot move habitat is perhaps a reason for declining numbers where habitats have become unsuitable.

We have built a series of vectors to allow the constitutive or lint-replate expression of foreign genes in plants. These vectors carry expression cassettes consisting of either the cauliflower mosaic virus 3% promoter or the pea rbcS-E9 promoter, a multiple cloning site derived from M 13um20, and the rbcS-E9 polyadenylation site. These cassettes have been ~~orporated into pBR322-based or RK2-based replicons to facilitate direct DNA uptake or Agrobacterium tumefaciem-mediated gene transfer. Their application for the expression of a bacterial gene is described.

We have built a series of vectors to allow the constitutive or lint-replate expression of foreign genes in plants. These vectors carry expression cassettes consisting of either the cauliflower mosaic virus 3% promoter or the pea rbcS-E9 promoter, a multiple cloning site derived from M 13um20, and the rbcS-E9 polyadenylation site. These cassettes have been ~~orporated into pBR322-based or RK2-based replicons to facilitate direct DNA uptake or Agrobacterium tumefaciem-mediated gene transfer. Their application for the expression of a bacterial gene is described.

We present a landslide susceptibility model for the Collazzone area, central Italy, and we propose a framework for evaluating the model reliability and prediction skill. The landslide susceptibility model was obtained through discriminant analysis of 46 thematic environmental variables and using the presence of shallow landslides obtained from a multi-temporal inventory map as the dependent variable for statistical analysis. By comparing the number of correctly and incorrectly classified mapping units, it is established that the model classifies 77.0% of 894 mapping units correctly. Model fitting performance is investigated by comparing the proportion of the study area in each probability class with the corresponding proportion of landslide area. We then prepare an ensemble of 350 landslide susceptibility models using the same landslide and thematic information but different numbers of mapping units. This ensemble is exploited to investigate the model reliability, including the role of the thematic variables used to construct the model, and the model sensitivity to changes in the input data. By studying the variation of the model's susceptibility estimate, the error associated with the susceptibility assessment for each mapping unit is determined. This result is shown on a map that complements the landslide susceptibility map. Prediction skill of the susceptibility model is then estimated by comparing the forecast with two recent event inventory maps. The susceptibility model is found capable of predicting the newly triggered landslides. A general framework for testing a susceptibility model is proposed, including a scheme for ranking the quality of the susceptibility assessment.

Exposure to alcohol in the embryonic mouse can lead to structural and neurophysiologic changes. The cause of these changes is poorly understood, but they are likely the result of numerous mechanisms. Here we investigate ethanol-induced alterations in gene expression in the fetal brain. Using complementary-DNA microarrays, we identified 25 genes that were down-regulated by prenatal ethanol exposure on days 7 and 9 of gestation. None were found to be up-regulated. Of those that were repressed, 6 (Timp4, Bmp15, Rnf25, Akt1, Tulp4, Dexras1) have been identified, and they are discussed here in the context of the developing fetus. The identified genes have been shown to be involved in cell proliferation, differentiation, and apoptosis, and they contribute to tissue growth and remodeling, as well as neuronal growth and survival. Microarray studies may be useful in the identification of a genetic marker for fetal alcohol syndrome, the discovery of novel pathways that may be involved in its origin, or both. (J Lab Clin Med 2005;145:47-54) Abbreviations: Akt1 ϭ thymoma viral protooncogene-1; BLAST ϭ Basic Local Alignment Research Tool, a public service offered by the National Centre for Biotechnology Information; Bmp15 ϭ bone morphogenic protein-15; Cy3 ϭ cyanine-3; Cy5 ϭ cyanine-5; Dexras1 ϭ dexamethasone-induced RAS-1; cDNA ϭ complementary DNA; dCTP ϭ deoxycytidine triphosphate; dNTP ϭ deoxyribonucleoside triphosphate; ECM ϭ extracellular matrix; EDTA ϭ ethylenediaminetetraacetate; EST ϭ expressed sequence tag; FAS ϭ fetal alcohol syndrome; FASD ϭ fetal alcohol spectrum disorder; FDR ϭ false-discovery rate; GD ϭ gestational day; Gdf9 ϭ growth-differentiation factor-9; GDP ϭ guanine diphosphate; GTP ϭ guanine triphosphate; MAP ϭ mitogen-activated protein; MMP ϭ matrix metalloproteinase; PCR ϭ polymerase chain reaction; Rnf25 ϭ ring-finger protein-25; SAM ϭ significance analysis of microarrays; SSC ϭ standard saline citrate; TGF-␤ ϭ transforming growth factor-␤; TIMP ϭ tissue-derived inhibitor of metalloproteinases; Timp4 ϭ tissue inhibitor of metalloproteinase-4; Tulp ϭ tubbylike protein; TUSP ϭ tubby-superfamily protein; tRNA ϭ transfer RNA A lcohol consumption during pregnancy often results in abnormal fetal development, both in human beings 1,2 and experimental animal models, 3-7 leading to FASD. The anomalies associated with full-blown FAS include pre-and postnatal growth retardation, distinct craniofacial dysmorphology, and deficits of the central nervous system. 2 Ethanol, which produces its most severe effects on the developing brain, causes a broad array of neuropathologic changes, including microcephaly, reduced volume of cerebral white matter and cerebellar hypoplasia. 8,9 Despite efforts to better understand the origin and molecular From the Division of Clinical Pharmacology and Toxicology and the Metabolism Program,

The localization of two cytoskeletal proteins was analysed in the cell bodies and processes of ganglionic neurons and small, intensely fluorescent cells of the parasympathetic cardiac ganglion of Necturus maculasw (mudpuppy). Antibodies against microtubule-associated protein-2 and against the highly phosphorylated isofonns of high and middle molecular weight neurohlament subunits were. used as somatodendritic and axonal markers, respectively. The ganglionic neurons, which usually have only one major process, and small, intensely fluorescent cells, which have several processes, showed distinctly different staining patterns with the two antibodies. In control and denervated ganglia, the ganglionic cell bodies and several hundred micrometers of the proximal processes were labeled with the antibody against microtubule-associated protein-2, whereas small, intensely fluorescent cells and processes showed a paucity of immunoreactivity. The neurofilarnent antibody labeled numerous axons in the ganglion but did not label the proximal part of the postganglionic process or small, intensely fluorescent cell processes. Denervation resulted in the presence of phosphorylated neurofilament subunit immunoreactivity in the soma and proximal process of the ganglionic neuron.

Smoothened is a member of the G-protein coupled receptor (GPCR) family responsible for the transduction of the Hedgehog signal to the intracellular effectors of the Hedgehog signaling pathway. Aberrant regulation of this receptor is implicated in many cancers but also in neurodegenerative disorders. Despite the pharmacological relevance of this receptor, very little is known about its functional mechanism and its physiological ligand. In order to characterize this receptor for basic and pharmacological interests, we developed the expression of human Smoothened in the yeast Saccharomyces cerevisiae and Smoothened was then purified. Using Surface Plasmon Resonance technology, we showed that human Smoothened was in a native conformational state and able to interact with its antagonist, the cyclopamine, both at the yeast plasma membrane and after purification. Thermostability assays on purified human Smoothened showed that this GPCR is relatively stable in the classical detergent dodecyl-β-d-maltoside (DDM). The fluorinated surfactant C8F17TAC, which has been proposed to be less aggressive towards membrane proteins than classical detergents, increased Smoothened thermostability in solution. Moreover, the replacement of a glycine by an arginine in the third intracellular loop of Smoothened coupled to the use of the fluorinated surfactant C8F17TAC during the mutant purification increased Smoothened thermostability even more. These data will be very useful for future crystallization assays and structural characterization of the human receptor Smoothened.

The mast cell is a central player in allergy and asthma. Activation of these cells induces the release of preformed inflammatory mediators localized in specialized granules and the de novo synthesis and secretion of cytokines, chemokines, and eicosanoids. The balance of engaging inhibitory and activatory cell-surface receptors on mast cells determines whether the cell becomes active on encountering a challenge. However, recent evidence suggests that, once activated, a mast cell's response is further regulated by the balance of both positive and negative intracellular molecular events that extend well beyond the traditional role of kinases and phosphatases. These functional responses are also carefully governed by other protein and lipid mediators that determine the rate and extent of the response. Molecules that have adaptor functions, modulate lipids, and provide synergistic signals add to the regulatory complexity. Considerable information has been obtained from the study of the high-affinity receptor for IgE (FcɛRI), and thus it is the major focus of this review. The unifying theme is that the regulatory steps mentioned herein are required for promoting effective responses while protecting against unwanted inflammatory responses.

Luteolin occurs as glycosylated forms in celery, green pepper, perilla leaf and camomile tea, and has been shown to possess antimutagenic, antitumorigenic, antioxidant and antiinflammatory properties. In this study, we have investigated the antiproliferable effect and its mechanism of luteolin on platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic vascular smooth muscle cells (VSMCs). Luteolin significantly inhibited PDGF-BB-induced proliferation and DNA synthesis of rat aortic VSMCs in a concentration-dependent manner. In addition, flow cytometry analysis of DNA content revealed blocking of the PDGF-BB-inducible cell cycle progression by luteolin. Pre-incubation of rat aortic VSMCs with luteolin significantly inhibited the PDGF-BB-induced extracellular signal-regulated kinase 1/2 (ERK1/2), Akt and phospholipase C (PLC)-γ1 activation as well as c-fos gene expression. Consisted with these findings, luteolin inhibited PDGF-Rβ phosphorylation induced by PDGF-BB in a concentration-dependent manner. These results suggest that the inhibitory effect of luteolin on the PDGF-BB-induced proliferation of rat aortic VSMCs may be mediated by blocking phosphorylation of PDGF-Rβ.

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This paper presents an intelligent GIS solution. The aim of this project is the realization of an open platform of services for the identification of persons that are in difficulty, in any location, by using Trimble Lasser SK II GPS receivers and Zapp terminals needed to assure the wireless Internet connection, based on CDMA technology and TCP/IP protocol.

Defects in macrophage function have been implicated in the establishment of Crohn's disease (CD). However, the response of macrophages from CD patients to live bacteria, particularly Mycobacterium avium subsp. paratuberculosis (MAP), has not been addressed. Considering MAP has long been associated to CD, our objective was to assess whether macrophages from CD patients showed impaired inflammatory response to infection by MAP comparing to M. avium subsp. avium (MA) and other live intestinal commensal bacteria. Human peripheral blood monocyte-derived macrophages were obtained from CD patients, ulcerative colitis (UC) patients and controls. Following in vitro infection with MAP, MA, Escherichia coli or Enterococcus faecalis, cytokine levels and cell surface receptor expression were evaluated at different time points. Macrophages from CD patients showed impaired TNF-α secretion in response to bacterial challenge, but augmented IL-23 secretion and preserved IL-12 secretion and CD-40 expression. In addition, CD macrophages showed low IL-10 secretion. Macrophages from IBD patients showed increased expression of TLR-2 and -4, unaffected by infection. Differences in cytokine secretion observed after bacterial challenge were not MAP-specific, as other bacteria (E. coli and MA) showed similar effects. Macrophages from UC patients showed a less compromised TNF-α synthesis in response to mycobacterial infection than CD macrophages, with increased constitutive IL-12 secretion, and preserved IL-10 secretion. The increased IL-23 levels in response to infection and decreased IL-10 production observed in macrophages from CD patients may contribute to the inflammatory exacerbation observed in those patients.

Broccoli (Brassica oleracea L. var. Italica) heads were packaged using 3 types of polypropylene films: macro-perforated (Ma-P), micro-perforated (Mi-P) and non-perforated (No-P), and then stored at 1 °C for 28 days to study the effects of modified atmosphere packaging (MAP) on the maintenance of quality and functional properties by comparison with non-wrapped heads. Results revealed that deterioration occurred quickly in control broccoli, manifested mainly by weight loss, yellowing, chlorophyll degradation and stem hardening. Also, a rapid decrease in total antioxidant activity (TAA), ascorbic acid and total phenolic compound concentration was observed. Conversely, in those heads packaged under MAP, especially for Mi-P and No-P, all changes related with loss of quality were significantly reduced and delayed with time. Additionally, TAA, ascorbic acid and total phenolic compounds remained almost unchanged during the whole period. Thus, broccoli packaged with Mi-P and No-P films had prolonged storability up to 28 days with high quality attributes and health-promoting compounds, this period being only 5 days in unwrapped control broccoli.

Numerous investigations have provided evidence that chicken products are a source of Listeria monocytogenes and Campylobacter jejuni. Different strategies applied in final products are needed to prevent consumers' contamination. In this work, the combination of modified atmosphere packaging (MAP) and protective culture to control the growth of freeze stressed and non-stressed L. monocytogenes and C. jejuni on fresh chicken meat burger was studied. Meat burgers were inoculated with L. monocytogenes, C. jejuni and Leuconostoc pseudomesenteroides PCK 18, as protective strain against L. monocytogenes. Prior to the addition of the protective culture, half of the L. monocytogenes and C. jejuniinoculated meat was frozen at − 18°C for 48 h to subject cells to stress. Following the addition of the protective culture, meat burgers were packaged in air or MAP (50% CO 2 /50% O 2 ) and stored under refrigeration conditions. L. monocytogenes counts were not reduced by the freezing temperature applied; however, the addition of Lc. pseudomesenteroides PCK 18 reduced its counts for 0.90 log cfu/g when chicken meat burgers were packaged under MAP. Furthermore, freezing stress was an effective strategy to reduce C. jejuni counts but only in combination with a high-O 2 MAP, it was completely eliminated. Chicken meat burgers' shelf-life under aerobic packaging conditions was reduced by the effect of freeze-thawing, while the use of MAP extended the product's shelf-life till 21 days. Therefore, the combination of freezing, protective culture and MAP could extend the shelf-life and enhance the food safety of this kind of chicken products.

Smoothness assumptions in traditional image expansion cause blurring of edges and other highfrequency content that can be perceptually disturbing. Previous edge-preserving approaches are either ad hoc, statistically untenable, or computationally unattractive. We propose a new edgedriven stochastic prior image model and obtain the maximum a posteriori (MAP) estimate under this model. The MAP estimate is computationally challenging since it involves the inversion of very large matrices. An efficient algorithm is presented for expansion by dyadic factors. The technique exploits diagonalization of convolutional operators under the Fourier transform, and the sparsity of our edge prior, to speed up processing. Visual and quantitative comparison of our technique with other popular methods demonstrates its potential and promise. Abstract-Smoothness assumptions in traditional image expansion cause blurring of edges and other high-frequency content that can be perceptually disturbing. Previous edge-preserving approaches are either ad hoc, statistically untenable, or computationally unattractive. We propose a new edge-driven stochastic prior image model and obtain the maximum a posteriori (MAP) estimate under this model. The MAP estimate is computationally challenging since it involves the inversion of very large matrices. An efficient algorithm is presented for expansion by dyadic factors. The technique exploits diagonalization of convolutional operators under the Fourier transform, and the sparsity of our edge prior, to speed up processing. Visual and quantitative comparison of our technique with other popular methods demonstrates its potential and promise.

Defects in macrophage function have been implicated in the establishment of Crohn's disease (CD). However, the response of macrophages from CD patients to live bacteria, particularly Mycobacterium avium subsp. paratuberculosis (MAP), has not been addressed. Considering MAP has long been associated to CD, our objective was to assess whether macrophages from CD patients showed impaired inflammatory response to infection by MAP comparing to M. avium subsp. avium (MA) and other live intestinal commensal bacteria. Human peripheral blood monocyte-derived macrophages were obtained from CD patients, ulcerative colitis (UC) patients and controls. Following in vitro infection with MAP, MA, Escherichia coli or Enterococcus faecalis, cytokine levels and cell surface receptor expression were evaluated at different time points. Macrophages from CD patients showed impaired TNF-␣ secretion in response to bacterial challenge, but augmented IL-23 secretion and preserved IL-12 secretion and CD-40 expression. In addition, CD macrophages showed low IL-10 secretion. Macrophages from IBD patients showed increased expression of TLR-2 and -4, unaffected by infection. Differences in cytokine secretion observed after bacterial challenge were not MAP-specific, as other bacteria (E. coli and MA) showed similar effects. Macrophages from UC patients showed a less compromised TNF-␣ synthesis in response to mycobacterial infection than CD macrophages, with increased constitutive IL-12 secretion, and preserved IL-10 secretion. The increased IL-23 levels in response to infection and decreased IL-10 production observed in macrophages from CD patients may contribute to the inflammatory exacerbation observed in those patients.

The influence of aniso-osmolarity on the activity of the MAP kinases Erk-1 and Erk-2 was studied in C6 glioma cells. Hypo-osmotic treatment (205 mosinol/l) led to an increased activity of Erk-1 and Erk-2 within 3 min, which became maximal at 10 min and returned to basal level within 120 min. In contrast, Erk activity was reduced under hyper-osmotic conditions (405 mosmol/1), compared to the normo-osmotic control (305 mosmol/ 1). Erk activation was accompanied by a mobility shift of Raf-1. Hypo-osmotic exposure increased the cytosolic Ca 2+ concentration (|Ca 2+ ]i). Absence of extracellular Ca 2+ largely abolished the ICa 2+ ]i response to hypo-osmolarity, whereas Erk activation following hypo-osmotic stimulation remained unaffected, suggesting a Ca 2+ independence of the osmosignalling pathway to the MAP kinases. Both the Ca 2+ response as well as the Erk activation following hypo-osmotic exposure were maintained in the presence of the phospholipase C inhibitor U73122.

The recovery of arousal after cardiac arrest (CA) is associated with evolution from electroencephalographic (EEG) burst-suppression to continuous activity. Orexin-A elicits arousal EEG during anesthetic burst-suppression. We hypothesized that orexin-A would improve arousal and EEG entropy after CA. Eighteen Wistar rats were subjected to 7-minute asphyxial CA and resuscitation. Rats were divided into treatment (n= 9) and control (n= 9) groups. Twenty minutes after resuscitation, the treatment group received 0.1 mL of 1 nM ...

Bayesian averaging (BA) over ensembles of decision models allows evaluation of the uncertainty of decisions that is of crucial importance for safety-critical applications such as medical diagnostics. The interpretability of the ensemble can also give useful information for experts responsible for making reliable decisions. For this reason, decision trees (DTs) are attractive decision models for experts. However, BA over such models makes an ensemble of DTs uninterpretable. In this paper, we present a new approach to probabilistic interpretation of Bayesian DT ensembles. This approach is based on the quantitative evaluation of uncertainty of the DTs, and allows experts to find a DT that provides a high predictive accuracy and confident outcomes. To make the BA over DTs feasible in our experiments, we use a Markov Chain Monte Carlo technique with a reversible jump extension. The results obtained from clinical data show that in terms of predictive accuracy, the proposed method outperforms the maximum a posteriori (MAP) method that has been suggested for interpretation of DT ensembles.

Introduction and objectives: To study the clinical course of children requiring continuous renal replacement therapy after cardiac surgery and to analyze factors associated with mortality. Methods: A prospective observational study was performed that included children requiring continuous renal replacement therapy after cardiac surgery. Univariate and multivariate analyses were performed to determine the influence of each factor on mortality. We compared these patients with other critically ill children requiring continuous renal replacement therapy. Results: Of 1650 children undergoing cardiac surgery, 81 (4.9%) required continuous renal replacement therapy, 65 of whom (80.2%) presented multiple organ failure. The children who started continuous renal replacement therapy after cardiac surgery had lower mean arterial pressure, lower urea and creatinine levels, and higher mortality (43%) than the other children on continuous renal replacement therapy (29%) (P = .05). Factors associated with mortality in the univariate analysis were age less than 12 months, weight under 10 kg, higher pediatric risk of mortality score, hypotension, lower urea and creatinine levels when starting continuous renal replacement therapy, and the use of hemofiltration. In the multivariate analysis, hypotension when starting continuous renal replacement therapy, pediatric risk of mortality scores equal to or greaterer than 21, and hemofiltration were associated with mortality. Conclusions: Although only a small percentage of children undergoing cardiac surgery required continuous renal replacement therapy, mortality among these patients was high. Hypotension and severity of illness when starting the technique and hemofiltration were factors associated with higher mortality.

This work presents a method to bridge the gap between intentional process modeling and business process modeling. The first represent the business objectives of an enterprise and the strategies used in order to achieve these objectives, while the second concentrate on the business processes. The proposed method uses MAP as an intentional modeling language and Business Process Modeling Notation (BPMN) as a modeling language for the business processes. We propose to translate the strategic goals expressed with MAP model into a BPMN process diagram. We show that an alignment of the intentional model (MAP) with BPMN can support the designers in transforming easily the strategic goals into business operational goals. We also show in this work, an example illustrating the use of our mapping.

Several studies from our group have indicated that the BNST play an important role in baroreflex modulation. However, the involvement of the BNST in the chemoreflex activity is unknown. Thus, in the present study, we investigated the effect of the local bed nucleus of stria terminalis (BNST) neurotransmission inhibition by bilateral microinjections of the non-selective synaptic blocker cobalt chloride (CoCl 2 ) on the cardiovascular responses to chemoreflex activation in rats. For this purpose, chemoreflex was activated with KCN (i.v.) before and after microinjections of CoCl 2 into the BNST. Reversible BNST inactivation produced no significant changes in the magnitude and durations of both pressor and bradycardic responses to intravenous KCN infusion. These findings suggesting that BNST neurotransmission have not influence on both sympathoexcitatory and parasympathoexcitatory components of the peripheral chemoreflex activation.

Multi-VMap is a compact framework from which plane graphs representing geographic maps at different levels of detail can be extracted. Its main feature is that the scale of the extracted map can be variable through its domain, while each entity maintains consistent combinatorial relations with the rest of entities represented in the map. The model is based on a set of operators, called updates, which modify the level of detail in a small portion of a map. The set of updates is partially ordered, and can therefore be represented as a Directed Acyclic Graph, which defines our multi-scale structure. An algorithm to extract a map at the required resolution is proposed, and a lower bound for the number of different maps which can be extracted from the model is given. The model supports map data processing operations (e.g., querying), as well as progressive and selective transmission of maps over a network.

Fresh-cut Amarillo melon was stored under passive modified atmosphere packaging (MAP) for 14 days at 5 C. Three commercial films were tested: microperforated polypropylene (MPP), bioriented PP (BPP), and oriented PP (OPP). As a control, a macroperforated PP film was used. The effect of a citric acid dip treatment (0.52 mM) on the quality of the melon pieces packaged in OPP film was also evaluated. A similar final gas composition (4 kPa O 2 plus 12-13 kPa CO 2 ) within packages was achieved by using the three tested films. This atmosphere was effective for maintaining sensorial quality and microbial safety and for avoiding weight loss and translucency. When citric acid dip was applied, a reduction of microbial counts, a low discoloration, an increase of lightness, and a general improvement of visual appearance were found. However, after 14 days of storage, neither MAP treatments nor citric acid dip were enough to avoid softening of at least 23%, although the highest value (32%) was found in control pieces. Taking into account safety, quality attributes and consumer acceptance, shelf life of fresh processed melon stored under the studied MAP conditions could be stabilised for 10 days.

The ErbB-3 receptor binding protein (Ebp1) is a member of the proliferation-associated 2G4 (PA2G4) family implicated in regulation of cell growth and differentiation. Here, we report the crystal structure of the human Ebp1 at 1.6 Å resolution. The protein has the conserved pita bread fold of methionine aminopeptidases, but without the characteristic enzymatic activity. Moreover, Ebp1 is known to interact with a number of proteins and RNAs involved in either transcription regulation or translation control. The structure provides insights in how Ebp1 discriminates between its different interaction partners.

Defects in macrophage function have been implicated in the establishment of Crohn's disease (CD). However, the response of macrophages from CD patients to live bacteria, particularly Mycobacterium avium subsp. paratuberculosis (MAP), has not been addressed. Considering MAP has long been associated to CD, our objective was to assess whether macrophages from CD patients showed impaired inflammatory response to infection by MAP comparing to M. avium subsp. avium (MA) and other live intestinal commensal bacteria. Human peripheral blood monocyte-derived macrophages were obtained from CD patients, ulcerative colitis (UC) patients and controls. Following in vitro infection with MAP, MA, Escherichia coli or Enterococcus faecalis, cytokine levels and cell surface receptor expression were evaluated at different time points. Macrophages from CD patients showed impaired TNF-␣ secretion in response to bacterial challenge, but augmented IL-23 secretion and preserved IL-12 secretion and CD-40 expression. In addition, CD macrophages showed low IL-10 secretion. Macrophages from IBD patients showed increased expression of TLR-2 and -4, unaffected by infection. Differences in cytokine secretion observed after bacterial challenge were not MAP-specific, as other bacteria (E. coli and MA) showed similar effects. Macrophages from UC patients showed a less compromised TNF-␣ synthesis in response to mycobacterial infection than CD macrophages, with increased constitutive IL-12 secretion, and preserved IL-10 secretion. The increased IL-23 levels in response to infection and decreased IL-10 production observed in macrophages from CD patients may contribute to the inflammatory exacerbation observed in those patients.

a b s t r a c t The innate immune system responds to inflammation, infection and injury by recruiting neutrophils and other leukocytes. These cells are able to leave the intravascular compartment in a process called leukocyte recruitment. This process involves several distinct steps: selectin-mediated rolling, firm adhesion via integrins, postarrest modifications including adhesion strengthening and leukocyte crawling and finally transmigration into tissue. Genetic defects affecting the different steps of the cascade can result in severe impairment in leukocyte recruitment. So far, three leukocyte adhesion deficiencies (LAD I-III) have been described in humans. These LADs are rare autosomal recessive inherited disorders and, although clinically distinct, exhibit several common features including recurrent bacterial infections and leukocytosis. In LAD-I, mutations within the ␤2-integrin gene result in a severe defect in ␤2 integrin-mediated firm leukocyte adhesion. Defects in the posttranslational fucosylation of selectin ligands dramatically reduce leukocyte rolling and lead to LAD-II. Finally, LAD-III, also known as LAD-I variant, is caused by impaired integrin activation due to mutations within the kindlin-3 gene. This review provides an overview on the molecular basis of leukocyte adhesion and its deficiencies.

Background: Urotensin-II (U-II) is a vasoactive peptide with diffuse expression in human cardiomyocyte and vascular smooth muscle cells. Recent studies have reported increased plasma levels of U-II in patients with congestive heart failure. Objective: We sought to determine the plasma levels of U-II in patients with acute coronary syndromes (ACS), stable coronary artery disease (CAD) and healthy controls. Methods: We prospectively measured plasma U-II levels in 54 patients with ACS, 51 patients with stable coronary disease and 29 healthy volunteers. Monoclonal antibodies against U-II were generated and plasma U-II levels were determined by radioimmunoassay from extracted venous samples. Results: ACS patients had significantly lower levels than patients with stable CAD and healthy controls (2.53 T 1.62 vs. 3.45 T 2.53 vs. 3.3 T 3.9 ng/ml, p = 0.008, respectively). In both ACS and stable CAD patients, we found a negative relationship between plasma U-II levels and systemic arterial pressures. The correlation coefficients for systolic and mean arterial pressure were À0.272, p = 0.006 and À0.209, p = 0.04, respectively. Conclusions: Plasma U-II levels were significantly decreased in patients with acute coronary syndromes and related negatively to systemic arterial pressures. This finding suggests a down-regulation of U-II expression in patients with acute coronary syndromes. Condensed abstract: Urotensin-II (U-II) is a vasoactive peptide with diffuse staining in human cardiomyocytes and vascular smooth muscle cells. We measured plasma U-II levels in patients with acute coronary syndromes (ACS), stable coronary artery disease (CAD) and healthy controls. We observed lower U-II levels in ACS patients and a negative correlation between U-II levels and systemic arterial pressure. This finding suggests a down-regulation of U-II expression in patients with ACS. D

Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli O157:H7 (EHEC) form characteristic lesions on infected mammalian cells called actin pedestals. Each of these two pathogens injects its own translocated intimin receptor (Tir) molecule into the plasma membranes of host cells. Interaction of translocated Tir with the bacterial outer membrane protein intimin is required to trigger the assembly of actin into focused pedestals beneath bound bacteria. Despite similarities between the Tir molecules and the host components that associate with pedestals, recent work indicates that EPEC and EHEC Tir are not functionally interchangeable. For EPEC, Tir-mediated binding of Nck, a host adaptor protein implicated in actin signaling, is both necessary and suf®cient to initiate actin assembly. In contrast, for EHEC, pedestals are formed independently of Nck, and require translocation of bacterial factors in addition to Tir to trigger actin signaling.

The problem of multiple testing is an important aspect of genome-wide association studies, and will become more important as marker densities increase. The problem has been tackled with permutation and false discovery rate procedures and with Bayes factors, but each approach faces difficulties that we briefly review. In the current context of multiple studies on different genotyping platforms, we argue for the use of truly genome-wide significance thresholds, based on all polymorphisms whether or not typed in the study. We approximate genome-wide significance thresholds in contemporary West African, East Asian and European populations by simulating sequence data, based on all polymorphisms as well as for a range of single nucleotide polymorphism (SNP) selection criteria. Overall we find that significance thresholds vary by a factor of 420 over the SNP selection criteria and statistical tests that we consider and can be highly dependent on sample size. We compare our results for sequence data to those derived by the HapMap Consortium and find notable differences which may be due to the small sample sizes used in the HapMap estimate. Genet. Epidemiol. 32:179-185, 2008.

ACF ϭ aorto-caval fistula CHF ϭ congestive heart failure eNOS ϭ endothelial NOS GFR ϭ glomerular filtration rate IAP ϭ intra-abdominal pressure L-NAME ϭ nitro-L-arginine methyl ester MAP ϭ mean arterial blood pressure NO ϭ nitric oxide NOS ϭ NO synthase PAH ϭ p-aminohippurate RBF ϭ renal blood flow RPF ϭ renal plasma flow U Na V ϭ urinary Na ϩ excretion U NO2 ϩ NO3 V ϭ urinary NO 2 and NO 3 excretion V ϭ urinary flow ).

Background: Inorganic nitrate (NO 3 − ) is a precursor of nitric oxide (NO) in the body and a large number of short-term studies with dietary nitrate supplementation in animals and humans show beneficial effects on cardiovascular health, exercise efficiency, host defense and ischemia reperfusion injury. In contrast, there is a long withstanding concern regarding the putative adverse effects of chronic nitrate exposure related to cancer and adverse hormonal effects. To address these concerns we performed in mice, a physiological and biochemical multi-analysis on the effects of long-term dietary nitrate supplementation. Design: 7 week-old C57BL/6 mice were put on a low-nitrate chow and at 20 weeks-old were treated with NaNO 3 (1 mmol/L) or NaCl (1 mmol/L, control) in the drinking water. The groups were monitored for weight gain, food and water consumption, blood pressure, glucose metabolism, body composition and oxygen consumption until one group was reduced to eight animals due to death or illness. At that point remaining animals were sacrificed and blood and tissues were analyzed with respect to metabolism, cardiovascular function, inflammation, and oxidative stress. Results: Animals were supplemented for 17 months before final sacrifice. Body composition, oxygen consumption, blood pressure, glucose tolerance were measured during the experiment, and vascular reactivity and muscle mitochondrial efficiency measured at the end of the experiment with no differences identified between groups. Nitrate supplementation was associated with improved insulin response, decreased plasma IL-10 and a trend towards improved survival. Conclusions: Long term dietary nitrate in mice, at levels similar to the upper intake range in the western society, is not detrimental.