Does an Integrated Care Intervention for COPD Patients Have Long-Term Effects on Quality of Life and Patient Activation? A Prospective, Open, Controlled Single-Center Intervention Study.
Implementation of the COPD-Home integrated disease management (IDM) intervention at discharge after hospitalizations for acute exacerbations of COPD (AECOPD) led to reduced hospital utilization during the following 24 months compared to the year prior to study start. To analyze the impact of the COPD-Home IDM intervention on health related quality of life, symptoms of anxiety and depression, and the degree of patient activation during 24 months of follow-up and to assess the association between these outcomes. A single center, prospective, open, controlled clinical study. Changes in The St. George Respiratory Questionnaire (SGRQ), the Hospital anxiety (HADS-A) and depression (HADS-D) and the patient activation measure (PAM) scores were compared between the patients in the integrated care group (ICG) and the usual care group (UCG) 6, 12 and 24 months after enrolment. The questionnaire response rate was 80-96%. There were no statistically significant differences in the change of the SGRQ scores between the groups during follow up. After 12 months of follow-up there was a trend towards a reduction in the mean HADS-A score in the ICG compared to the UCG. The HADS-D scores remained stable in the ICG compared with an increasing trend in the UCG. Clinically significant difference in the PAM score was achieved only in the ICG, 6.7 (CI95% 0.7 to 7.5) compared to 3.6 (CI95% -1.4 to 8.6) in the UCG. In a logistic regression model a higher HADS-D score and current smoking significantly increased the odds for a low PAM score. The COPD-Home IDM intervention did not result in any statistically significant changes in mean SGRQ, HADS-A, HADS- D or PAM scores during the 24 months of follow-up. The ID number for the study in the Clinical.Trials.gov registration system is 17417. ClinicalTrials.gov Identifier: NCT 00702078.


A boundary for histone acetylation allows distinct expression patterns of the Ad4BP/SF-1 and GCNF loci in adrenal cortex cells.
Ad4BP/SF-1 is a nuclear receptor whose expression is restricted to tissues involved in steroid hormone synthesis such as the adrenal cortex and gonads. Recent sequence data analysis has shown that the Ad4BP/SF-1 gene is located only 13kb downstream of the last exon of the neighboring GCNF gene that is expressed in some neurons and gonadal germ cells. Despite the close proximity of the two genes, regulatory elements from one do not interfere with the transcription of the neighboring gene, resulting in distinct expression patterns of Ad4BP/SF-1 and GCNF. This observation has led to the prediction that an insulator element must exist between the two loci to establish independent transcription units. We performed DNase I hypersensitivity assays on the adrenal cortex cell line, Y-1, to test for the existence of an insulator. Three hypersensitive sites were identified in the region spanning 2.1kb between the last exon of GCNF and the first exon of Ad4BP/SF-1. The most upstream site contains a binding site for CTCF, a known insulator protein, while the other sites are predicted to associate with the nuclear matrix. Chromatin immunoprecipitation analysis using anti-acetylated histone H3 and H4 antibodies showed a discontinuous pattern of histone H3 and H4 acetylation upstream of these sites. Our data suggest that the chromatin architecture specialized by CTCF and the nuclear matrix contribute to the distinct pattern of transcriptional regulation of these genes.


Gallotannin inhibits the expression of chemokines and inflammatory cytokines in A549 cells.
Tannins are plant-derived water-soluble polyphenols with wide-ranging biological activities. The mechanisms underlying the anti-inflammatory effect of tannins are not fully understood and may be the result of inhibition of poly(ADP-ribose) (PAR) glycohydrolase (PARG), the main catabolic enzyme of PAR metabolism. Therefore, we set out to investigate the mechanism of the anti-inflammatory effect of gallotannin (GT) in A549 cells with special regard to the role of poly(ADP-ribosyl)ation. Using an inflammation-focused low-density array and reverse transcription-polymerase chain reaction, we found that GT suppressed the expression of most cytokines and chemokines in cytokine-stimulated A549 cells, whereas the PARP inhibitor PJ-34 only inhibited few transcripts. Activation of the transcription factors, nuclear factor kappaB (NF-kappaB) and activator protein 1 (AP-1), was blocked by GT, whereas PJ-34 only suppressed NF-kappaB activation but not AP-1 activation. GT also inhibited IkappaB phosphorylation and nuclear translocation of NF-kappaB, but PJ-34 had no effect on these upstream events. In the AP-1 pathway, GT treatment, even in the absence of cytokines, caused maximal phosphorylation of c-Jun N-terminal kinase and c-Jun. GT also caused a low-level phosphorylation of p38, extracellular signal-regulated kinases 1 and 2, activating transcription factor2, and cAMP-response element-binding protein but inhibited cytokine-induced phosphorylation of these kinases and transcription factors. GT inhibited protein phosphatases 1 and 2A, which may explain the increased phosphorylation of mitogen-activated protein kinase and their substrates. GT exerted potent antioxidant effect but failed to cause PAR accumulation. In summary, the potent inhibitory effects of GT on the transcription of cytokine and chemokine genes are probably not related to PARG inhibition. Inhibition of AP-1 activation and upstream signaling events may be responsible for the effects of GT.


Mn(II) Oxidation by the Multicopper Oxidase Complex Mnx: A Binuclear Activation Mechanism.
The bacterial protein complex Mnx contains a multicopper oxidase (MCO) MnxG that, unusually, catalyzes the two-electron oxidation of Mn(II) to MnO2 biomineral, via a Mn(III) intermediate. Although Mn(III)/Mn(II) and Mn(IV)/Mn(III) reduction potentials are expected to be high, we find a low reduction potential, 0.38 V (vs Normal Hydrogen Electrode, pH 7.8), for the MnxG type 1 Cu2+, the electron acceptor. Indeed the type 1 Cu2+ is not reduced by Mn(II) in the absence of molecular oxygen, indicating that substrate oxidation requires an activation step. We have investigated the enzyme mechanism via electronic absorption spectroscopy, using chemometric analysis to separate enzyme-catalyzed MnO2 formation from MnO2 nanoparticle aging. The nanoparticle aging time course is characteristic of nucleation and particle growth; rates for these processes followed expected dependencies on Mn(II) concentration and temperature, but exhibited different pH optima. The enzymatic time course is sigmoidal, signaling an activation step, prior to turnover. The Mn(II) concentration and pH dependence of a preceding lag phase indicates weak Mn(II) binding. The activation step is enabled by a pKa > 8.6 deprotonation, which is assigned to Mn(II)-bound H2O; it induces a conformation change (consistent with a high activation energy, 106 kJ/mol) that increases Mn(II) affinity. Mnx activation is proposed to decrease the Mn(III/II) reduction potential below that of type 1 Cu(II/I) by formation of a hydroxide-bridged binuclear complex, Mn(II)(μ-OH)Mn(II), at the substrate site. Turnover is found to depend cooperatively on two Mn(II) and is enabled by a pKa 7.6 double deprotonation. It is proposed that turnover produces a Mn(III)(μ-OH)2Mn(III) intermediate that proceeds to the enzyme product, likely Mn(IV)(μ-O)2Mn(IV) or an oligomer, which subsequently nucleates MnO2 nanoparticles. We conclude that Mnx exploits manganese polynuclear chemistry in order to facilitate an otherwise difficult oxidation reaction, as well as biomineralization. The mechanism of the Mn(III/IV) conversion step is elucidated in an accompanying paper .


Experimental Insight into the Structural and Functional Roles of the 'Black' and 'Gray' Clusters in Recoverin, a Calcium Binding Protein with Four EF-Hand Motifs.
Recently, we have found that calcium binding proteins of the EF-hand superfamily (i.e., a large family of proteins containing helix-loop-helix calcium binding motif or EF-hand) contain two types of conserved clusters called cluster I ('black' cluster) and cluster II ('grey' cluster), which provide a supporting scaffold for the Ca2+ binding loops and contribute to the hydrophobic core of the EF-hand domains. Cluster I is more conservative and mostly incorporates aromatic amino acids, whereas cluster II includes a mix of aromatic, hydrophobic, and polar amino acids of different sizes. Recoverin is EF-hand Ca2+-binding protein containing two 'black' clusters comprised of F35, F83, Y86 (N-terminal domain) and F106, E169, F172 (C-terminal domain) as well as two 'gray' clusters comprised of F70, Q46, F49 (N-terminal domain) and W156, K119, V122 (C-terminal domain). To understand a role of these residues in structure and function of human recoverin, we sequentially substituted them for alanine and studied the resulting mutants by a set of biophysical methods. Under metal-free conditions, the 'black' clusters mutants (except for F35A and E169A) were characterized by an increase in the α-helical content, whereas the 'gray' cluster mutants (except for K119A) exhibited the opposite behavior. By contrast, in Ca2+-loaded mutants the α-helical content was always elevated. In the absence of calcium, the substitutions only slightly affected multimerization of recoverin regardless of their localization (except for K119A). Meanwhile, in the presence of calcium mutations in N-terminal domain of the protein significantly suppressed this process, indicating that surface properties of Ca2+-bound recoverin are highly affected by N-terminal cluster residues. The substitutions in C-terminal clusters generally reduced thermal stability of recoverin with F172A ('black' cluster) as well as W156A and K119A ('gray' cluster) being the most efficacious in this respect. In contrast, the mutations in the N-terminal clusters caused less pronounced differently directed changes in thermal stability of the protein. The substitutions of F172, W156, and K119 in C-terminal domain of recoverin together with substitution of Q46 in its N-terminal domain provoked significant but diverse changes in free energy associated with Ca2+ binding to the protein: the mutant K119A demonstrated significantly improved calcium binding, whereas F172A and W156A showed decrease in the calcium affinity and Q46A exhibited no ion coordination in one of the Ca2+-binding sites. The most of the N-terminal clusters mutations suppressed membrane binding of recoverin and its inhibitory activity towards rhodopsin kinase (GRK1). Surprisingly, the mutant W156A aberrantly activated rhodopsin phosphorylation regardless of the presence of calcium. Taken together, these data confirm the scaffolding function of several cluster-forming residues and point to their critical role in supporting physiological activity of recoverin.


Tumor Formation in Interspecific Hybrids of Lilium.
Tumors were formed on germinating seeds of a hybrid of Lilium speciosum "Album" x L. auratum. Ferulic acid and the glucose ester were isolated from both pericarp and seeds. Tumors were induced on excised embryos only when the embryos were grown on nutrient agar containing ferulic acid. Tumors may result from the limited amount of glycosylation and from detoxification of ferulic acid.


Structure and mechanism of human diacylglycerol O-acyltransferase 1.
Diacylglycerol O-acyltransferase 1 (DGAT1) synthesizes triacylglycerides and is required for dietary fat absorption and fat storage in humans1. DGAT1 belongs to the membrane-bound O-acyltransferase (MBOAT) superfamily, members of which are found in all kingdoms of life and are involved in the acylation of lipids and proteins2,3. How human DGAT1 and other mammalian members of the MBOAT family recognize their substrates and catalyse their reactions is unknown. The absence of three-dimensional structures also hampers rational targeting of DGAT1 for therapeutic purposes. Here we present the cryo-electron microscopy structure of human DGAT1 in complex with an oleoyl-CoA substrate. Each DGAT1 protomer has nine transmembrane helices, eight of which form a conserved structural fold that we name the MBOAT fold. The MBOAT fold in DGAT1 forms a hollow chamber in the membrane that encloses highly conserved catalytic residues. The chamber has separate entrances for each of the two substrates, fatty acyl-CoA and diacylglycerol. DGAT1 can exist as either a homodimer or a homotetramer and the two forms have similar enzymatic activity. The N terminus of DGAT1 interacts with the neighbouring protomer and these interactions are required for enzymatic activity.


Temporal sparseness of the premotor drive is important for rapid learning in a neural network model of birdsong.
Sparse neural codes have been widely observed in cortical sensory and motor areas. A striking example of sparse temporal coding is in the song-related premotor area high vocal center (HVC) of songbirds: The motor neurons innervating avian vocal muscles are driven by premotor nucleus robustus archistriatalis (RA), which is in turn driven by nucleus HVC. Recent experiments reveal that RA-projecting HVC neurons fire just one burst per song motif. However, the function of this remarkable temporal sparseness has remained unclear. Because birdsong is a clear example of a learned complex motor behavior, we explore in a neural network model with the help of numerical and analytical techniques the possible role of sparse premotor neural codes in song-related motor learning. In numerical simulations with nonlinear neurons, as HVC activity is made progressively less sparse, the minimum learning time increases significantly. Heuristically, this slowdown arises from increasing interference in the weight updates for different synapses. If activity in HVC is sparse, synaptic interference is reduced, and is minimized if each synapse from HVC to RA is used only once in the motif, which is the situation observed experimentally. Our numerical results are corroborated by a theoretical analysis of learning in linear networks, for which we derive a relationship between sparse activity, synaptic interference, and learning time. If songbirds acquire their songs under significant pressure to learn quickly, this study predicts that HVC activity, currently measured only in adults, should also be sparse during the sensorimotor phase in the juvenile bird. We discuss the relevance of these results, linking sparse codes and learning speed, to other multilayered sensory and motor systems.


Evoking Photothermy by Capturing Intramolecular Bond Stretching Vibration-Induced Dark-State Energy.
Development of highly effective approaches to desirable photothermal conversion agents is particularly valuable. Herein, we report a concept, namely, bond stretching vibration-induced photothermy, that serves as a mechanism to construct advanced photothermal conversion agents. As a proof-of-concept, two compounds (DCP-TPA and DCP-PTPA) with donor-acceptor (D-A) structures were synthesized. The bond stretching vibration of the pyrazine-containing unit in these molecules is vigorous and insensitive to the external environmental restraint, which efficiently transforms the absorbed photons to dark-state heat energy. The nanoparticles (NPs) of DCP-TPA and DCP-PTPA show rather high photothermal conversion efficiency (52% and 59%) and stronger photoacoustic (PA) signal than commercial methylene blue and reported high-performance semiconducting polymer nanoparticles. The DCP-PTPA NPs perform better than DCP-TPA NPs in terms of photothermal conversion, PA signal production, and in vivo PA tumor imaging because of the increased bond stretching vibration in the former molecule.


Differential activity-dependent, homeostatic plasticity of two neocortical inhibitory circuits.
Chronic changes in neuronal activity homeostatically regulate excitatory circuitry. However, little is known about how activity regulates inhibitory circuits or specific inhibitory neuron types. Here, we examined the activity-dependent regulation of two neocortical inhibitory circuits--parvalbumin-positive (Parv+) and somatostatin-positive (Som+)--using paired recordings of synaptically coupled neurons. Action potentials were blocked for 5 days in slice culture, and unitary synaptic connections among inhibitory/excitatory neuron pairs were examined. Chronic activity blockade caused similar and distinct changes between the two inhibitory circuits. First, increases in intrinsic membrane excitability and excitatory synaptic drive in both inhibitory subtypes were consistent with the homeostatic regulation of firing rate of these neurons. On the other hand, inhibitory synapses originating from these two subtypes were differentially regulated by activity blockade. Parv+ unitary inhibitory postsynaptic current (uIPSC) strength was decreased while Som+ uIPSC strength was unchanged. Using short-duration stimulus trains, short-term plasticity for both unitary excitatory postsynaptic current (uEPSCs) and uIPSCs was unchanged in Parv+ circuitry while distinctively altered in Som+ circuitry--uEPSCs became less facilitating and uIPSCs became more depressing. In the context of recurrent inhibition, these changes would result in a frequency-dependent shift in the relative influence of each circuit. The functional changes at both types of inhibitory connections appear to be mediated by increases in presynaptic release probability and decreases in synapse number. Interestingly, these opposing changes result in decreased Parv+-mediated uIPSCs but balance out to maintain normal Som+-mediated uIPSCs. In summary, these results reveal that inhibitory circuitry is not uniformly regulated by activity levels and may provide insight into the mechanisms of both normal and pathological neocortical plasticity.


Spanish Guidelines for the use of targeted deep sequencing in myelodysplastic syndromes and chronic myelomonocytic leukaemia.
The landscape of medical sequencing has rapidly changed with the evolution of next generation sequencing (NGS). These technologies have contributed to the molecular characterization of the myelodysplastic syndromes (MDS) and chronic myelomonocytic leukaemia (CMML), through the identification of recurrent gene mutations, which are present in >80% of patients. These mutations contribute to a better classification and risk stratification of the patients. Currently, clinical laboratories include NGS genomic analyses in their routine clinical practice, in an effort to personalize the diagnosis, prognosis and treatment of MDS and CMML. NGS technologies have reduced the cost of large-scale sequencing, but there are additional challenges involving the clinical validation of these technologies, as continuous advances are constantly being made. In this context, it is of major importance to standardize the generation, analysis, clinical interpretation and reporting of NGS data. To that end, the Spanish MDS Group (GESMD) has expanded the present set of guidelines, aiming to establish common quality standards for the adequate implementation of NGS and clinical interpretation of the results, hoping that this effort will ultimately contribute to the benefit of patients with myeloid malignancies.


Inhibition of bone resorption in culture by inhibitors of thiol proteinases.
Leupeptin, antipain, tosyl-lysylchloromethane (Tos-Lys-CH2Cl) and benzyloxy-carbonylphenylalanylalanyldiazomethane (Z-Phe-Ala-CHN2) inhibit reversibly the resorption induced by parathyroid hormone or heparin in cultured mouse bones. Leupeptin and antipain do not affect collagenase production and activity or the enhanced secretion of beta-glucuronidase induced by the bone-resorbing agents. They might thus act by a direct (extracellular?) inhibition of lysosomal thiol proteinases.


Can manipulation of the ratios of essential fatty acids slow the rapid rate of postmenopausal bone loss?
The rapid rate of postmenopausal bone loss is mediated by the inflammatory cytokines interleukin-1, interleukin-6, and tumor necrosis factor alpha. Dietary supplementation with fish oil, flaxseeds, and flaxseed oil in animals and healthy humans significantly reduces cytokine production while concomitantly increasing calcium absorption, bone calcium, and bone density. Possibilities may exist for the therapeutic use of the omega-3 fatty acids, as supplements or in the diet, to blunt the increase of the inflammatory bone resorbing cytokines produced in the early postmenopausal years, in order to slow the rapid rate of postmenopausal bone loss. Evidence also points to the possible benefit of gamma-linolenic acid in preserving bone density.


Splicing of a divergent subclass of AT-AC introns requires the major spliceosomal snRNAs.
AT-AC introns constitute a minor class of eukaryotic pre-mRNA introns, characterized by 5'-AT and AC-3' boundaries, in contrast to the 5'-GT and AG-3' boundaries of the much more prevalent conventional introns. In addition to the AT-AC borders, most known AT-AC introns have highly conserved 5' splice site and branch site sequence elements of 7-8 nt. Intron 6 of the nucleolar P120 gene and intron 2 of the SCN4A voltage-gated skeletal muscle sodium channel are AT-AC introns that have been shown recently to be processed via a unique splicing pathway involving several minor U snRNAs. Interestingly, intron 21 of the same SCN4A gene and the corresponding intron 25 of the SCN5A cardiac muscle sodium channel gene also have 5'-AT and AC-3' boundaries, but they have divergent 5' splice site and presumptive branch site sequences. Here, we report the accurate in vitro processing of these two divergent AT-AC introns and show that they belong to a functionally distinct subclass of AT-AC introns. Splicing of these introns does not require U12, U4atac, and U6atac snRNAs, but instead requires the major spliceosomal snRNAs U1, U2, U4, U5, and U6. Previous studies showed that G --> A mutation at the first position and G --> C mutation at the last position of a conventional yeast or mammalian GT-AG intron suppress each other in vivo, suggesting that the first and last bases participate in an essential non-Watson-Crick interaction. Our results show that such introns, hereafter termed AT-AC II introns, occur naturally and are spliced by a mechanism distinct from that responsible for processing of the apparently more common AT-AC I introns.


Antisocial and Prosocial Behavior in Sport: The Role of Motivational Climate, Basic Psychological Needs, and Moral Disengagement.
The purpose of this investigation was to examine whether the relationships between contextual factors and basic psychological needs were related to antisocial and prosocial behavior in sport. A two-study project employing Bayesian path analysis was conducted with competitive athletes (Study 1, n = 291; Study 2, n = 272). Coach and teammate autonomy-supportive climates had meaningful direct relations with need satisfaction and prosocial behavior. Coach and teammate controlling climates had meaningful direct relations with antisocial behavior. Need satisfaction was both directly and indirectly related with both prosocial and antisocial behavior, whereas moral disengagement was directly and indirectly related with antisocial behavior. Overall, these findings reflected substantial evidence from the literature on self-determination theory that autonomy-supportive motivational climates are important environmental influences for need satisfaction, and are important correlates of prosocial behavior in sport, whereas controlling coach and teammate climates, along with moral disengagement, were important correlates of antisocial behavior in sport.


The Simon task and aging: does acute moderate exercise influence cognitive control?
This study aimed to investigate the influence of an acute bout of moderate exercise and examine the potential lasting improvements over time in young and old adults within the same experimental paradigm over a 2-h testing period. The study was designed to assess the efficiency of selective control and the propensity to make fast impulsive reactions through the analyses of the percentage of correct responses (CAF) and the magnitude of the interference effect (delta curve) as a function of the latency of the response. Twelve young (23 ± 2 yr) and 12 old (63 ± 2 yr) volunteers performed the Simon task while cycling (30 min of cycling at 65% of age-predicted HRmax) and after exercise cessation (post 5 min, post 35 min, and post 65 min). Results showed that exercise did not alter cognitive control. The benefit on reaction time performance was evident for both age groups and persisted after cessation for 15-20 min. Distributional analyses showed that younger people have a higher propensity to commit impulsive errors during exercise, which was not evident in older adults. Older adults adopted more cautious strategies, especially when the risk to commit an error was elevated. Despite the larger mean interference effect compared to younger adults, the pattern of the delta curves attests to the existence of an efficient cognitive control in older people. This study illustrates the effectiveness of distributional analyses and supports the idea that exercise-induced facilitation on cognitive performance can be realized across the lifespan. Future investigations should explore whether accumulated bouts of acute exercise could display an aggregate cognitive benefit, which may significantly affect independent functioning in older adults.


The Effects of Concentrative Meditation on the Electroencephalogram in Novice Meditators.
Following investigations into the benefits of meditation on psychological health and well-being, research is now seeking to understand the mechanisms underlying these outcomes. This study aimed to identify natural alpha and theta frequency components during eyes-closed resting and concentrative meditation states and examined their differences within and between two testing sessions. Novice meditators had their EEG recorded during eyes-closed resting and concentrative meditation conditions, before and after engaging in a brief daily concentrative meditation practice for approximately one-month. Separate frequency Principal Components Analyses (f-PCA) yielded four spectral components of interest, congruent between both conditions and sessions: Delta-Theta-Alpha, Low Alpha, High Alpha, and Alpha-Beta. While all four components showed some increase in the meditation condition at the second session, only Low Alpha (∼9.5-10.0 Hz) showed similar increases while resting. These findings support the use of f-PCA as a novel method of data analysis in the investigation of psychophysiological states in meditation.


Effect of hypoxia on cerebral metabolites measured by proton nuclear magnetic resonance spectroscopy in rats.
Proton nuclear magnetic resonance spectroscopy is a unique method to monitor noninvasively the concentrations of cerebral metabolites. N-Acetyl-L-aspartate, the concentration of which is assumed to be stable during hypoxia, has been used to form ratios with lactate. To determine the stability of the signal from N-acetyl-L-aspartate, we used a model of graded hypoxia in rats to monitor the percentage changes from baseline of the peak heights for lactate, lipids, and N-acetyl-L-aspartate. Anesthetized adult rats were exposed sequentially to 15% and 10% O2 while proton nuclear magnetic resonance spectra were collected with a surface coil in a 7-T 89-mm-bore spectrometer. Brain lactate concentration was either increased by feeding or infusion of glucose (n = 9) or lowered by fasting (n = 7). After death the brains were removed and frozen, and the water- and lipid-soluble compounds were extracted to identify the origin of the signals. We analyzed the data both as the percentage change from baseline for heights of the lactate (1.33 ppm), lipids (1.5 ppm), and N-acetyl-L-aspartate (2.02 ppm) peaks and as the ratios of heights of the 1.33 and 2.02 and the 1.5 and 2.02 ppm peaks. Both hypoxic episodes caused a 45% decrease from baseline in the 2.02 ppm peak. During the second hypoxic episode, the 1.33:2.02 ppm peak height ratio increased significantly in hyperglycemic rats (p less than 0.05) but was unchanged in hypoglycemic rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Morphological and molecular characterization of Phytophthora glovera sp. nov. from tobacco in Brazil.
A root rot disease of cultivated tobacco called yellow stunt has been observed in the burley tobacco production areas of Brazil since the early 1990s. Root infecting fungi and straminipiles were isolated from the roots of diseased tobacco plants, including a semi-papillate, homothallic, slow growing Phytophthora species. Pathogenicity trials confirmed that Phytophthora sp. caused root rot and stunting of burley and flue-cured tobaccos. Morphological characteristics of the asexual and sexual stages of this organism did not match any reported Phytophthora species and were very different from the widely known tobacco black shank pathogen P. nicotianae. Phylogenetic analysis based on sequences of the internal transcribed spacer rDNA, β-tubulin and translation elongation factor 1-α regions indicated that this organism represents a previously unreported Phytophthora species that is significantly supported in clade 2 and most closely related to P. capsici. However P. glovera differs from P. capsici in a number of morphological characters, most significantly P. glovera is homothallic and produces both paragynous and amphigynous antheridia while P. capsici is heterothallic and produces only amphigynous antheridia. In this paper we confirmed pathogenicity of this species on tobacco and describe the morphological and molecular characteristics of Phytophthora glovera sp. nov.


Nanomaterial building blocks based on spider silk-oligonucleotide conjugates.
Self-assembling protein nanofibrils are promising structures for the "bottom-up" fabrication of bionanomaterials. Here, the recombinant protein eADF4(C16), a variant of Araneus diadematus dragline silk ADF4, which self-assembles into nanofibrils, and short oligonucleotides were modified for site-specific azide-alkyne coupling. Corresponding oligonuleotide-eADF4(C16) "click" conjugates were hybridized in linear or branched fashion according to the designed complementarities of the DNA moieties. Self-assembly properties of higher ordered structures of the spider silk-DNA conjugates were dominated by the silk component. Assembled β-sheet rich conjugate fibrils were similar in appearance to fibrils of unmodified eADF4(C16) but enabled the specific attachment of neutravidin-modified gold nanoparticles on their surface directed by complementary biotin-oligonucleotides, providing the basis for functionalization of such conjugates.


Ultra-pH-sensitive nanoprobe library with broad pH tunability and fluorescence emissions.
pH is an important physiological parameter that plays a critical role in cellular and tissue homeostasis. Conventional small molecular pH sensors (e.g., fluorescein, Lysosensor) are limited by broad pH response and restricted fluorescent emissions. Previously, we reported the development of ultra-pH-sensitive (UPS) nanoprobes with sharp pH response using fluorophores with small Stokes shifts (<40 nm). In this study, we expand the UPS design to a library of nanoprobes with operator-predetermined pH transitions and wide fluorescent emissions (400-820 nm). A copolymer strategy was employed to fine tune the hydrophobicity of the ionizable hydrophobic block, which led to a desired transition pH based on standard curves. Interestingly, matching the hydrophobicity of the monomers was critical to achieve a sharp pH transition. To overcome the fluorophore limitations, we introduced copolymers conjugated with fluorescence quenchers (FQs). In the micelle state, the FQs effectively suppressed the emission of fluorophores regardless of their Stokes shifts and further increased the fluorescence activation ratios. As a proof of concept, we generated a library of 10 nanoprobes each encoded with a unique fluorophore. The nanoprobes cover the entire physiologic range of pH (4-7.4) with 0.3 pH increments. Each nanoprobe maintained a sharp pH transition (on/off < 0.25 pH) and high fluorescence activation ratio (>50-fold between on and off states). The UPS library provides a useful toolkit to study pH regulation in many pathophysiological indications (e.g., cancer, lysosome catabolism) as well as establishing tumor-activatable systems for cancer imaging and drug delivery.


Characterisation of the Manduca sexta sperm proteome: Genetic novelty underlying sperm composition in Lepidoptera.
The application of mass spectrometry based proteomics to sperm biology has greatly accelerated progress in understanding the molecular composition and function of spermatozoa. To date, these approaches have been largely restricted to model organisms, all of which produce a single sperm morph capable of oocyte fertilisation. Here we apply high-throughput mass spectrometry proteomic analysis to characterise sperm composition in Manduca sexta, the tobacco hornworm moth, which produce heteromorphic sperm, including one fertilisation competent (eupyrene) and one incompetent (apyrene) sperm type. This resulted in the high confidence identification of 896 proteins from a co-mixed sample of both sperm types, of which 167 are encoded by genes with strict one-to-one orthology in Drosophila melanogaster. Importantly, over half (55.1%) of these orthologous proteins have previously been identified in the D. melanogaster sperm proteome and exhibit significant conservation in quantitative protein abundance in sperm between the two species. Despite the complex nature of gene expression across spermatogenic stages, a significant correlation was also observed between sperm protein abundance and testis gene expression. Lepidopteran-specific sperm proteins (e.g., proteins with no homology to proteins in non-Lepidopteran taxa) were present in significantly greater abundance on average than those with homology outside the Lepidoptera. Given the disproportionate production of apyrene sperm (96% of all mature sperm in Manduca) relative to eupyrene sperm, these evolutionarily novel and highly abundant proteins are candidates for possessing apyrene-specific functions. Lastly, comparative genomic analyses of testis-expressed, ovary-expressed and sperm genes identified a concentration of novel sperm proteins shared amongst Lepidoptera of potential relevance to the evolutionary origin of heteromorphic spermatogenesis. As the first published Lepidopteran sperm proteome, this whole-cell proteomic characterisation will facilitate future evolutionary genetic and developmental studies of heteromorphic sperm production and parasperm function. Furthermore, the analyses presented here provide useful annotation information regarding sex-biased gene expression, novel Lepidopteran genes and gene function in the male gamete to complement the newly sequenced and annotated Manduca genome.


Probing rotational wave-packet dynamics with the structural minimum in high-order harmonic spectra.
We investigate the alignment-dependent high-order harmonic spectrum generated from nonadiabatically aligned molecules around the first half rotational revival. It is found that the evolution of the molecular alignment is encoded in the structural minima. To reveal the relation between the molecular alignment and the structural minimum in the high-order harmonic spectrum, we perform an analysis based on the two-center interference model. Our analysis shows that the structural minimum position depends linearly on the inverse of the alignment parameter <cos2θ>. This linear relation indicates the possibility of probing the rotational wave-packet dynamics by measuring the spectral minima.


Species richness and trophic diversity increase decomposition in a co-evolved food web.
Ecological communities show great variation in species richness, composition and food web structure across similar and diverse ecosystems. Knowledge of how this biodiversity relates to ecosystem functioning is important for understanding the maintenance of diversity and the potential effects of species losses and gains on ecosystems. While research often focuses on how variation in species richness influences ecosystem processes, assessing species richness in a food web context can provide further insight into the relationship between diversity and ecosystem functioning and elucidate potential mechanisms underpinning this relationship. Here, we assessed how species richness and trophic diversity affect decomposition rates in a complete aquatic food web: the five trophic level web that occurs within water-filled leaves of the northern pitcher plant, Sarracenia purpurea. We identified a trophic cascade in which top-predators--larvae of the pitcher-plant mosquito--indirectly increased bacterial decomposition by preying on bactivorous protozoa. Our data also revealed a facultative relationship in which larvae of the pitcher-plant midge increased bacterial decomposition by shredding detritus. These important interactions occur only in food webs with high trophic diversity, which in turn only occur in food webs with high species richness. We show that species richness and trophic diversity underlie strong linkages between food web structure and dynamics that influence ecosystem functioning. The importance of trophic diversity and species interactions in determining how biodiversity relates to ecosystem functioning suggests that simply focusing on species richness does not give a complete picture as to how ecosystems may change with the loss or gain of species.


Evaluation of salivary biomarker profiles following non-surgical management of chronic periodontitis.
Soluble toll-like receptor-2 (sTLR-2) and cytokines in saliva were assessed as clinical markers for chronic periodontitis in a longitudinal study. Unstimulated whole saliva was collected from 20 periodontally healthy individuals and 20 patients with chronic periodontitis at diagnosis and at 1 and 6 weeks following scaling and root planing (SRP). Biomarkers including the cytokines (IL-4, IL-6, IL-10, and IL-17), sTLR-2, and sCD14 in saliva were measured by enzyme-linked immunosorbent assay. Mann-Whitney U-test and Student's t-test were used to determine the significance between healthy and chronic periodontitis groups and that between pre- and post-SRP samples, respectively. Salivary sTLR-2, IL-17, and IL-10 levels were significantly lower and those of sCD14, IL-6, and IL-4 were significantly higher in patients with chronic periodontitis as compared with healthy controls. Furthermore, sTLR-2 and IL-4 in saliva reached levels comparable to those of healthy individuals at 6-week re-evaluation visit, implicating a correlation of the two markers with the disease process. Our data suggest that salivary sTLR-2 is a potential prognostic or maintenance marker for chronic periodontitis. The observed variability of salivary cytokines is consistent with the role of these cytokines in the progression of chronic periodontitis.


Predictors of relapse in problem gambling: a prospective cohort study.
To explore the variation of predictors of relapse in treatment and support seeking gamblers. A prospective cohort study with 158 treatment and support seeking problem gamblers in South Australia. Key measures were selected using a consensus process with international experts in problem gambling and related addictions. The outcome measures were Victorian Gambling Screen (VGS) and behaviours related to gambling. Potential predictors were gambling related cognitions and urge, emotional disturbance, social support, sensation seeking traits, and levels of work and social functioning. Mean age of participants was 44 years (SD = 12.92 years) and 85 (54 %) were male. Median time for participants enrollment in the study was 8.38 months (IQR = 2.57 months). Patterns of completed measures for points in time included 116 (73.4 %) with at least a 3 month follow-up. Using generalised mixed-effects regression models we found gambling related urge was significantly associated with relapse in problem gambling as measured by VGS (OR 1.29; 95 % CI 1.12-1.49) and gambling behaviours (OR 1.16; 95 % CI 1.06-1.27). Gambling related cognitions were also significantly associated with VGS (OR 1.06; 95 % CI 1.01-1.12). There is consistent association between urge to gamble and relapse in problem gambling but estimates for other potential predictors may have been attenuated because of methodological limitations. This study also highlighted the challenges presented from a cohort study of treatment and support seeking problem gamblers.


Safety and immunogenicity of 2009 pandemic influenza A H1N1 vaccines in China: a multicentre, double-blind, randomised, placebo-controlled trial.
The current influenza pandemic calls for a safe and effective vaccine. We assessed the safety and immunogenicity of eight formulations of 2009 pandemic influenza A H1N1 vaccine produced by ten Chinese manufacturers. In this multicentre, double-blind, randomised trial, 12 691 people aged 3 years or older were recruited in ten centres in China. In each centre, participants were stratified by age and randomly assigned by a random number table to receive one of several vaccine formulations or placebo. The study assessed eight formulations: split-virion formulation containing 7.5 microg, 15 microg, or 30 microg haemagglutinin per dose, with or without aluminium hydroxide adjuvant, and whole-virion formulation containing 5 microg or 10 microg haemagglutinin per dose, with adjuvant. All formulations were produced from the reassortant strain X-179A (A/California/07/2009-A/PR/8/34). We analysed the safety (adverse events), immunogenicity (geometric mean titre [GMT] of haemagglutination inhibition antibody), and seroprotection (GMT >or=1:40) of the formulations. Analysis was by per protocol. Two sites registered their trial with ClinicalTrials.gov, numbers NCT00956111 and NCT00975572. The other eight studies were registered with the State Food and Drug Administration of China. 12 691 participants received the first dose on day 0, and 12 348 participants received the second dose on day 21. The seroprotection rate 21 days after the first dose of vaccine ranged from 69.5% (95% CI 65.9-72.8) for the 7.5 microg adjuvant split-virion formulation to 92.8% (91.9-93.6) for the 30 microg non-adjuvant split-virion formulation. The seroprotection rate was 86.5% (796 of 920; 84.1-88.7) in recipients of one dose of the 7.5 microg non-adjuvant split-virion vaccine compared with 9.8% (140 of 1432; 8.3-11.4) in recipients of placebo (p<0.0001). One dose of the 7.5 microg non-adjuvant split-virion vaccine induced seroprotection in 178 of 232 children (aged 3 years to <12 years; 76.7%, 70.7-82.0), 211 of 218 adolescents (12 years to <18 years; 96.8%, 93.5-98.7), 289 of 323 adults (18-60 years; 89.5%, 85.6-92.6), and 118 of 147 adults older than 60 years (80.3%, 72.9-86.4), meeting the European Union's licensure criteria for seroprotection in all age-groups. In children, a second dose of the 7.5 microg formulation increased the seroprotection rate to 97.7% (215 of 220, 94.8-99.3). Adverse reactions were mostly mild or moderate, and self-limited. Severe adverse effects occurred in 69 (0.6%, 0.5-0.8) recipients of vaccine compared with one recipient (0.1%, 0-0.2) of placebo. The most common severe adverse reaction was fever, which occurred in 25 (0.22%; 0.14-0.33) recipients of vaccine after the first dose and four (0.04%; 0.01-0.09) recipients of vaccine after the second dose compared with no recipients of placebo after either dose. One dose of non-adjuvant split-virion vaccine containing 7.5 microg haemagglutinin could be promoted as the formulation of choice against 2009 pandemic influenza A H1N1 for people aged 12 years or older. In children (aged <12 years), two 7.5 mug doses might be needed. Sinovac Biotech, Hualan Biological Bacterin, China National Biotec Group, Beijing Tiantan Biological Products, Changchun Institute of Biological Products, Changchun Changsheng Life Sciences, Jiangsu Yanshen Biological Technology Stock, Zhejiang Tianyuan Bio-Pharmaceutical, Lanzhou Institute of Biological Products, Shanghai Institute of Biological Products, and Dalian Aleph Biomedical.


Antithrombin: anti-inflammatory properties and clinical applications.
Many humoral and cellular components participate in bidirectional communication between the coagulation and inflammation pathways. Natural anticoagulant proteins, including antithrombin (AT), tissue factor pathway inhibitor, and protein C, suppress proinflammatory mediators. Conversely, inflammation blunts anticoagulant activity and, when uncontrolled, promotes systemic inflammation-induced coagulation, such as those that occur in disseminated intravascular coagulation and severe sepsis. This review discusses the mechanisms of action and clinical use of AT concentrate in critically ill patients and in the settings of perioperative anticoagulation management for surgery and obstetrics. AT is a serine protease inhibitor with broad anticoagulant activity and potent anti-inflammatory properties. In clinical conditions associated with hereditary or acquired AT deficiency, administration of AT concentrate has been shown to restore proper haemostasis and attenuate inflammation. Of note, AT modulates inflammatory responses not only by inhibiting thrombin and other clotting factors that induce cytokine activity and leukocyte-endothelial cell interaction, but also by coagulation-independent effects, including direct interaction with cellular mediators of inflammation. An increasing body of evidence suggests that AT concentrate may be a potential therapeutic agent in certain clinical settings associated with inflammation. In addition to the well-known anticoagulation properties of AT for the treatment of hereditary AT deficiency, AT also possesses noteworthy anti-inflammatory properties that could be valuable in treating acquired AT deficiency, which often result in thrombotic states associated with an inflammatory component.


The clinimetric qualities of patient-assessed instruments for measuring chronic ankle instability: a systematic review.
The assessment of outcomes from the patient's perspective becomes more recognized in health care. Also in patients with chronic ankle instability, the degree of present impairments, disabilities and participation problems should be documented from the perspective of the patient. The decision about which patient-assessed instrument is most appropriate for clinical practice should be based upon systematic reviews. Only rating scales constructed for patients with acute ligament injuries were systematically reviewed in the past. The aim of this study was to review systematically the clinimetric qualities of patient-assessed instruments designed for patients with chronic ankle instability. A computerized literature search of Medline, Embase, Cinahl, Web of Science, Sport Discus and the Cochrane Controlled Trial Register was performed to identify eligible instruments. Two reviewers independently evaluated the clinimetric qualities of the selected instruments using a criteria list. The inter-observer reliability of both the selection procedure and the clinimetric evaluation was calculated using modified kappa coefficients. The inter-observer reliability of the selection procedure was excellent (k = .86). Four instruments met the eligibility criteria: the Ankle Joint Functional Assessment Tool (AJFAT), the Functional Ankle Outcome Score (FAOS), the Foot and Ankle Disability Index (FADI) and the Functional Ankle Ability Measure (FAAM). The inter-observer reliability of the quality assessment was substantial to excellent (k between .64 and .88). Test-retest reliability was demonstrated for the FAOS, the FADI and the FAAM but not for the AJFAT. The FAOS and the FAAM met the criteria for content validity and construct validity. For none of the studied instruments, the internal consistency was sufficiently demonstrated. The presence of floor- and ceiling effects was assessed for the FAOS but ceiling effects were present for all subscales. Responsiveness was demonstrated for the AJFAT, FADI and the FAAM. Only for the FAAM, a minimal clinical important difference (MCID) was presented. The FADI and the FAAM can be considered as the most appropriate, patient-assessed tools to quantify functional disabilities in patients with chronic ankle instability. The clinimetric qualities of the FAAM need to be further demonstrated in a specific population of patients with chronic ankle instability.


Structure elucidation and biosynthetic gene cluster analysis of caniferolides A-D, new bioactive 36-membered macrolides from the marine-derived Streptomyces caniferus CA-271066.
Bioassay-guided isolation based on the antifungal activity of a culture broth of the marine-derived actinomycete Streptomyces caniferus CA-271066 led to the discovery of new 36-membered polyol macrolides, caniferolides A-D (1-4). Their connectivity was determined by spectroscopic methods including ESITOF-MS and 1D/2D NMR. The relative stereochemistry of each stereocluster in these compounds was established using NOE analysis, the universal database method and J-based configuration analysis, further assisted by comparisons with NMR data of structurally related macrolides. Genome sequencing followed by detailed bioinformatics analysis led to the identification of the corresponding biosynthetic gene cluster and allowed the prediction of the stereochemical outcome of their biosynthesis, confirming the relative stereochemistry of each stereocluster already determined by NMR and establishing their stereochemical relationship, ultimately rendering the absolute configuration of all chiral centers. Furthermore, based on our results and already published data, it has been possible to derive the complete absolute configuration of the related macrolides PM100117 and PM100118, astolides A and B, and deplelides A and B. Caniferolides A-D have shown pronounced antifungal activity against Candida albicans and Aspergillus fumigatus alongside antiproliferative activity against five human tumoral cell lines.


Acetaminophen poisoning: an evidence-based consensus guideline for out-of-hospital management.
The objective of this guideline is to assist poison center personnel in the appropriate out-of-hospital triage and initial management of patients with suspected ingestions of acetaminophen. An evidence-based expert consensus process was used to create this guideline. This guideline applies to ingestion of acetaminophen alone and is based on an assessment of current scientific and clinical information. The expert consensus panel recognizes that specific patient care decisions may be at variance with this guideline and are the prerogative of the patient and the health professionals providing care. The panel's recommendations follow. These recommendations are provided in chronological order of likely clinical use. The grade of recommendation is provided in parentheses. 1) The initial history obtained by the specialist in poison information should include the patient's age and intent (Grade B), the specific formulation and dose of acetaminophen, the ingestion pattern (single or multiple), duration of ingestion (Grade B), and concomitant medications that might have been ingested (Grade D). 2) Any patient with stated or suspected self-harm or who is the recipient of a potentially malicious administration of acetaminophen should be referred to an emergency department immediately regardless of the amount ingested. This referral should be guided by local poison center procedures (Grade D). 3) Activated charcoal can be considered if local poison center policies support its prehospital use, a toxic dose of acetaminophen has been taken, and fewer than 2 hours have elapsed since the ingestion (Grade A). Gastrointestinal decontamination could be particularly important if acetylcysteine cannot be administered within 8 hours of ingestion. Acute, single, unintentional ingestion of acetaminophen: 1) Any patient with signs consistent with acetaminophen poisoning (e.g., repeated vomiting, abdominal tenderness in the right upper quadrant or mental status changes) should be referred to an emergency department for evaluation (Grade D). 2) Patients less than 6 years of age should be referred to an emergency department if the estimated acute ingestion amount is unknown or is 200 mg/kg or more. Patients can be observed at home if the dose ingested is less than 200 mg/kg (Grade B). 3) Patients 6 years of age or older should be referred to an emergency department if they have ingested at least 10 g or 200 mg/kg (whichever is lower) or when the amount ingested is unknown (Grade D). 4) Patients referred to an emergency department should arrive in time to have a stat serum acetaminophen concentration determined at 4 hours after ingestion or as soon as possible thereafter. If the time of ingestion is unknown, the patient should be referred to an emergency department immediately (Grade D). 5) If the initial contact with the poison center occurs more than 36 hours after the ingestion and the patient is well, the patient does not require further evaluation for acetaminophen toxicity (Grade D). Repeated supratherapeutic ingestion of acetaminophen (RSTI): 1) Patients under 6 years of age should be referred to an emergency department immediately if they have ingested: a) 200 mg/kg or more over a single 24-hour period, or b) 150 mg/kg or more per 24-hour period for the preceding 48 hours, or c) 100 mg/kg or more per 24-hour period for the preceding 72 hours or longer (Grade C). 2) Patients 6 years of age or older should be referred to an emergency department if they have ingested: a) at least 10 g or 200 mg/kg (whichever is less) over a single 24-hour period, or b) at least 6 g or 150 mg/kg (whichever is less) per 24-hour period for the preceding 48 hours or longer. In patients with conditions purported to increase susceptibility to acetaminophen toxicity (alcoholism, isoniazid use, prolonged fasting), the dose of acetaminophen considered as RSTI should be greater than 4 g or 100 mg/kg (whichever is less) per day (Grade D). 3) Gastrointestinal decontamination is not needed (Grade D). Other recommendations: 1) The out-of-hospital management of extended-release acetaminophen or multi-drug combination products containing acetaminophen is the same as an ingestion of acetaminophen alone (Grade D). However, the effects of other drugs might require referral to an emergency department in accordance with the poison center's normal triage criteria. 2) The use of cimetidine as an antidote is not recommended (Grade A).


Cause and prevention of electrosurgical injuries in laparoscopy.
Electrosurgical injuries occur during laparoscopic operations and are potentially serious. The overall incidence of recognized injuries is between one and two patients per 1,000 operations. The majority go unrecognized at the time of the electrical insult and commonly present three to seven days afterward with fever and pain in the abdomen. Since these injuries appear late the pathophysiology remains speculative. This article reviewed the physics of electrosurgery and provides the surgeon with an insight to the mechanisms responsible in each type of injury. In addition, a comprehensive search of the world literature has reviewed all articles on the topic. The main causes of electrosurgical injuries are: inadvertent touching or grasping of tissue during current application, direct coupling between a portion of intestine and a metal probe that is touching the activated probe, insulation breaks in the electrodes, direct sparking to the intestine from the diathermy probe, and current passage to the intestine from recently coagulated, electrically isolated tissue. The majority of injuries, not surprisingly, are caused by monopolar diathermy. Bipolar diathermy is safer and should be used in preference to monopolar diathermy, especially in anatomically crowded areas. An awareness of the hazards of diathermy together with an understanding of the mechanisms of injury should enable the surgeon to dissect tissue and to achieve hemostasis, while at the same time decreasing the risk of serious complications to the patient.


Geo-Climatic Changes and Apomixis as Major Drivers of Diversification in the Mediterranean Sea Lavenders (Limonium Mill.).
The Mediterranean realm, comprising the Mediterranean and Macaronesian regions, has long been recognized as one of the world's biodiversity hotspots, owing to its remarkable species richness and endemism. Several hypotheses on biotic and abiotic drivers of species diversification in the region have been often proposed but rarely tested in an explicit phylogenetic framework. Here, we investigate the impact of both species-intrinsic and -extrinsic factors on diversification in the species-rich, cosmopolitan Limonium, an angiosperm genus with center of diversity in the Mediterranean. First, we infer and time-calibrate the largest Limonium phylogeny to date. We then estimate ancestral ranges and diversification dynamics at both global and regional scales. At the global scale, we test whether the identified shifts in diversification rates are linked to specific geological and/or climatic events in the Mediterranean area and/or asexual reproduction (apomixis). Our results support a late Paleogene origin in the proto-Mediterranean area for Limonium, followed by extensive in situ diversification in the Mediterranean region during the late Miocene, Pliocene, and Pleistocene. We found significant increases of diversification rates in the "Mediterranean lineage" associated with the Messinian Salinity Crisis, onset of Mediterranean climate, Plio-Pleistocene sea-level fluctuations, and apomixis. Additionally, the Euro-Mediterranean area acted as the major source of species dispersals to the surrounding areas. At the regional scale, we infer the biogeographic origins of insular endemics in the oceanic archipelagos of Macaronesia, and test whether woodiness in the Canarian Nobiles clade is a derived trait linked to insular life and a biotic driver of diversification. We find that Limonium species diversity on the Canary Islands and Cape Verde archipelagos is the product of multiple colonization events followed by in situ diversification, and that woodiness of the Canarian endemics is indeed a derived trait but is not associated with a significant shift to higher diversification rates. Our study expands knowledge on how the interaction between abiotic and biotic drivers shape the uneven distribution of species diversity across taxonomic and geographical scales.


Nutrigenomics, beta-cell function and type 2 diabetes.
The present investigation was designed to investigate the accuracy and precision of lactate measurement obtained with contemporary biosensors (Chiron Diagnostics, Nova Biomedical) and standard enzymatic photometric procedures (Sigma Diagnostics, Abbott Laboratories, Analyticon). Measurements were performed in vitro before and after the stepwise addition of 1 molar sodium lactate solution to samples of fresh frozen plasma to systematically achieve lactate concentrations of up to 20 mmol/l. Precision of the methods investigated varied between 1% and 7%, accuracy ranged between 2% and -33% with the variability being lowest in the Sigma photometric procedure (6%) and more than 13% in both biosensor methods. Biosensors for lactate measurement provide adequate accuracy in mean with the limitation of highly variable results. A true lactate value of 6 mmol/l was found to be presented between 4.4 and 7.6 mmol/l or even with higher difference. Biosensors and standard enzymatic photometric procedures are only limited comparable because the differences between paired determinations presented to be several mmol. The advantage of biosensors is the complete lack of preanalytical sample preparation which appeared to be the major limitation of standard photometry methods.


Estimating model-adjusted risks, risk differences, and risk ratios from complex survey data.
There is increasing interest in estimating and drawing inferences about risk or prevalence ratios and differences instead of odds ratios in the regression setting. Recent publications have shown how the GENMOD procedure in SAS (SAS Institute Inc., Cary, North Carolina) can be used to estimate these parameters in non-population-based studies. In this paper, the authors show how model-adjusted risks, risk differences, and risk ratio estimates can be obtained directly from logistic regression models in the complex sample survey setting to yield population-based inferences. Complex sample survey designs typically involve some combination of weighting, stratification, multistage sampling, clustering, and perhaps finite population adjustments. Point estimates of model-adjusted risks, risk differences, and risk ratios are obtained from average marginal predictions in the fitted logistic regression model. The model can contain both continuous and categorical covariates, as well as interaction terms. The authors use the SUDAAN software package (Research Triangle Institute, Research Triangle Park, North Carolina) to obtain point estimates, standard errors (via linearization or a replication method), confidence intervals, and P values for the parameters and contrasts of interest. Data from the 2006 National Health Interview Survey are used to illustrate these concepts.


Long-term effects of tungsten carbide (WC) nanoparticles in pelagic and benthic aquatic ecosystems.
As the production and usage of nanomaterials are increasing so are the concerns related to the release of the material into nature. Tungsten carbide (WC) is widely used for its hard metal properties, although its use, in for instance tyre studs, may result in nano-sized particles ending up in nature. Here, we evaluate the potential long-term exposure effects of WC nanoparticles on a pelagic (Daphnia magna) and a benthic (Asellus aquaticus) organism. No long-term effects were observed in the benthic system with respect to population dynamics or ecosystem services. However, long-term exposure of D. magna resulted in increased time to first reproduction and, if the particles were resuspended, strong effects on survival and reproductive output. Hence, the considerable differences in acute vs. long-term exposure studies revealed here emphasize the need for more long-term studies if we are to understand the effects of nanoparticles in natural systems.


Association of achieved dialysis dose with mortality in the hemodialysis study: an example of "dose-targeting bias".
In the intention-to-treat analysis of the Hemodialysis Study, all-cause mortality did not differ significantly between the high versus standard hemodialysis dose groups. The association of mortality with delivered dose within each of the two randomized treatment groups was examined, and implications for observational studies were considered. Time-dependent Cox regression was used to relate the relative risk (RR) for mortality to the running mean of the achieved equilibrated Kt/V (eKt/V) over the preceding 4 mo. eKt/V was categorized by quintiles within each dose group. Analyses were controlled for case-mix factors and baseline anthropometric volume. Within each randomized dose group, mortality was elevated markedly when achieved eKt/V was in the lowest quintile (RR, 1.93; 95% confidence interval [CI], 1.40 to 2.66; P < 0.0001 in the standard-dose group; RR, 2.04; 95% CI, 1.50 to 2.76; P < 0.0001 in the high-dose group; RR relative to the middle quintiles). The mortality rate in the lowest eKt/V quintile of the high-dose group was higher than in the full standard-dose group (RR, 1.59; 95% CI, 1.29 to 1.96; P < 0.0001). Each 0.1 eKt/V unit below the group median was associated with a 58% higher mortality in the standard-dose group (P < 0.001) and a 37% higher mortality in the high-dose group (P < 0.001). The magnitude of these dose-mortality effects was seven- to 12-fold higher than the upper limit of the 95% CI from the intention-to-treat analysis. The effects were attenuated in lagged analyses but did not disappear. When dialysis dose is targeted closely, as under the controlled conditions of the Hemodialysis Study, patients with the lowest achieved dose relative to their target dose experience markedly increased mortality, to a degree that is not compatible with a biologic effect of dose. The possibility of similar (albeit smaller) biases should be considered when analyzing observational data sets relating mortality to achieved dose of dialysis.


Study on the interaction between the 1,4,5,8-naphthalene diimide-spermine conjugate (NDIS) and DNA using a spectroscopic approach and molecular docking.
The interaction of herring sperm DNA with the 1,4,5,8-naphthalene diimide-spermine conjugate (NDIS) was studied by UV/vis absorption, fluorescence and CD spectroscopic methods. Compared with the 1,8-naphthalimide-spermidine conjugate (NIS), the values of KSV (quenching constant) and Kb (binding constant) of NDIS were larger, and the hypochromic effect in the UV/vis spectra and the quenching effect in the fluorescence of NDIS were more significant. The interaction mode between NDIS and DNA was mainly groove binding. The fluorescence experiments at varying temperatures showed that the binding process of NDIS and DNA was static, as both hydrogen bonds and hydrophobic forces played a major role in the binding of NDIS and DNA. The CD spectrum indicated that NDIS caused a conformational change, like the B to A-DNA transition, and the tests using KI and NaCl and 1H NMR spectroscopy indicated that NDIS was not a classical DNA inserter. All the results demonstrated that both the polyamine side chain and the aromatic rings affect the process of NDIS binding to DNA, which is thus obviously different from that of NIS. The conclusion was confirmed by the in silico molecular docking experiments.


An IoT Solution for Online Monitoring of Anesthetics in Human Serum Based on an Integrated Fluidic Bioelectronic System.
In this paper, we present the design, the implementation and the validation of a novel Internet of Things (IoT) drug monitoring system for the online continuous and simultaneous detection of two main anesthetics, e.g., propofol and paracetamol, in undiluted human serum. The described full system consists of a custom-built electronic Raspberry Pi (RPi) based Printed Circuit Board (PCB) that drives and reads out the signal from an electrochemical sensing platform integrated into a fluidic system. Thanks to the Polydimethylsiloxane (PDMS) fluidic device, the analyzed sample is automatically fluxed on the sensing site. The IoT network is supported by a Cloud system, which allows the doctor to control and share all the patient's data through a dedicated Android application and a smart watch. The validation closes with the first ever demonstration that our system successfully works for the simultaneous monitoring of propofol and paracetamol in undiluted human serum by measuring the concentration trends of these two drugs in fluxing conditions over time.


Listeria monocytogenes encodes a functional ESX-1 secretion system whose expression is detrimental to in vivo infection.
Bacterial pathogenicity deeply depends on the ability to secrete virulence factors that bind specific targets on host cells and manipulate host responses. The Gram-positive bacterium Listeria monocytogenes is a human foodborne pathogen that remains a serious public health concern. To transport proteins across its cell envelope, this facultative intracellular pathogen engages a set of specialized secretion systems. Here we show that L. monocytogenes EGDe uses a specialized secretion system, named ESX-1, to secrete EsxA, a homolog of the virulence determinants ESAT-6 and EsxA of Mycobacterium tuberculosis and Staphylococcus aureus, respectively. Our data show that the L. monocytogenes ESX-1 secretion system and its substrates are dispensable for bacterial invasion and intracellular multiplication in eukaryotic cell lines. Surprisingly, we found that the EssC-dependent secretion of EsxA has a detrimental effect on L. monocytogenes in vivo infection.


Computer Modeling of Clonal Dominance: Memory-Anti-Naïve and Its Curbing by Attrition.
Experimental and computational studies have revealed that T-cell cross-reactivity is a widespread phenomenon that can either be advantageous or detrimental to the host. In particular, detrimental effects can occur whenever the clonal dominance of memory cells is not justified by their infection-clearing capacity. Using an agent-based model of the immune system, we recently predicted the "memory anti-naïve" phenomenon, which occurs when the secondary challenge is similar but not identical to the primary stimulation. In this case, the pre-existing memory cells formed during the primary infection may be rapidly deployed in spite of their low affinity and can actually prevent a potentially higher affinity naïve response from emerging, resulting in impaired viral clearance. This finding allowed us to propose a mechanistic explanation for the concept of "antigenic sin" originally described in the context of the humoral response. However, the fact that antigenic sin is a relatively rare occurrence suggests the existence of evolutionary mechanisms that can mitigate the effect of the memory anti-naïve phenomenon. In this study we use computer modeling to further elucidate clonal dominance and the memory anti-naïve phenomenon, and to investigate a possible mitigating factor called attrition. Attrition has been described in the experimental and computational literature as a combination of competition for space and apoptosis of lymphocytes via type-I interferon in the early stages of a viral infection. This study systematically explores the relationship between clonal dominance and the mechanism of attrition. Our results suggest that attrition can indeed mitigate the memory anti-naïve effect by enabling the emergence of a diverse, higher affinity naïve response against the secondary challenge. In conclusion, modeling attrition allows us to shed light on the nature of clonal interaction and dominance.


Emerging Treatments for Malignant Pleural Mesothelioma: Where Are We Heading?
Malignant pleural mesothelioma (MPM) is an uncommon but aggressive and treatment resistant neoplasm with low survival rates. In the last years we assisted to an exponential growth in the appreciation of mesothelioma pathobiology, leading several new treatments to be investigated both in the early stage of the disease and in the advanced setting. In particular, expectations are now high that immunotherapy will have a leading role in the next years. However, caution is required as results from phase II studies in MPM were often not replicated in larger, randomized, phase III trials. In this review, we describe the most promising emerging therapies for the treatment of MPM, discussing the biological rationale underlying their development as well as the issues surrounding clinical trial design and proper selection of patients for every treatment.


Colonization of Phaseolus vulgaris nodules by Agrobacterium-like strains.
Non-nodulating Agrobacterium-like strains identified among root nodule isolates of common bean were labeled with gusA, a reporter gene encoding beta-glucuronidase (GUS). Bean plants were then co-inoculated with an infective Rhizobium strain and labeled transconjugants of Agrobacterium-like strains. Blue staining of nodules showed that Agrobacterium-like strains were able to colonize these symbiotic organs. Isolation and characterization by restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes revealed a mixed population of Rhizobium and Agrobacterium-like strains in all nodules showing GUS activity. PCR amplification of the nifH gene and nodulation tests did not show any evidence of acquisition of symbiotic gene by lateral transfer from Rhizobium to Agrobacterium-like strains. Moreover, these strains were able to invade mature nodules. Based on sequencing of the 16S rRNA gene, one of these Agrobacterium-like strains showed 99.4% sequence similarity with Agrobacterium bv. 1 reference strains and 99% similarity with an Agrobacterium bv. 1 strain isolated from Acacia mollisima in Senegal. Agrobacterium tumefaciens C58 and the disarmed variant AT123 did not show any ability to colonize nodules. Co-inoculation of bean seeds with Agrobacterium and Rhizobium strains did not enhance nodulation and plant yield under controlled conditions.


South Africa's COVID-19 Tracing Database: Risks and rewards of which doctors should be aware.
In response to the COVID-19 pandemic, South Africa (SA) has established a Tracing Database, collecting both aggregated and individualised mobility and locational data on COVID-19 cases and their contacts. There are compelling public health reasons for this development, since the database has the potential to assist with policy formulation and with contact tracing. While potentially demonstrating the rapid facilitation through technology of an important public service, the Tracing Database does, however, infringe immediately upon constitutional rights to privacy and heightens the implications of ethical choices facing medical professionals. The medical community should be aware of this surveillance innovation and the risks and rewards it raises. To deal with some of these risks, including the potential for temporary rights- infringing measures to become permanent, there are significant safeguards designed into the Tracing Database, including a strict duration requirement and reporting to a designated judge. African states including SA should monitor this form of contact tracing closely, and also encourage knowledge-sharing among cross-sectoral interventions such as the Tracing Database in responding to the COVID-19 pandemic.


A person-centred approach in nursing: Validity and reliability of the Carer Support Needs Assessment Tool.
The Carer Support Needs Assessment Tool (CSNAT) was developed for use among family caregivers in palliative care for assessment of their support needs. The purpose of this study was to translate and evaluate the validity and reliability of the CSNAT in a sample of Swedish family caregivers and nurses in a palliative care context. Data for this validation study was collected during 2016 in the context of palliative home care in two larger Swedish cities. The study was conducted in three stages to reach conceptual, semantic, operational and measurement equivalence between the original UK version and the Swedish version. Stage I consisted of translation to Swedish. In Stage II, cognitive interviews were performed with 8 family caregivers and 10 nurses. Data were analyzed based on relevance, clarity and sensitivity. In Stage III, the CSNAT and related self-rating measures (caregiver burden, preparedness for caregiving and quality of life) were completed by 118 family caregivers. Data quality, construct validity and test-retest reliability were evaluated. The CSNAT items were considered relevant and useful to identify areas of support needs. The Swedish CSNAT showed sound psychometric properties with satisfactory data quality and few problems with missing data across items (1.8%-6.1%). All items except one correlated as expected (rho>0.3) with caregiver burden, supporting construct validity. All items had satisfactory test-retest reliability (κw=0.45-0.75). This study further adds to the validity of the CSNAT and shows in addition that it is reliable and stable for use among family caregivers in palliative care.


Both fallopian tube and ovarian surface epithelium are cells-of-origin for high-grade serous ovarian carcinoma.
The cell-of-origin of high grade serous ovarian carcinoma (HGSOC) remains controversial, with fallopian tube epithelium (FTE) and ovarian surface epithelium (OSE) both considered candidates. Here, by using genetically engineered mouse models and organoids, we assessed the tumor-forming properties of FTE and OSE harboring the same oncogenic abnormalities. Combined RB family inactivation and Tp53 mutation in Pax8 + FTE caused Serous Tubal Intraepithelial Carcinoma (STIC), which metastasized rapidly to the ovarian surface. These events were recapitulated by orthotopic injection of mutant FTE organoids. Engineering the same genetic lesions into Lgr5 + OSE or OSE-derived organoids also caused metastatic HGSOC, although with longer latency and lower penetrance. FTE- and OSE-derived tumors had distinct transcriptomes, and comparative transcriptomics and genomics suggest that human HGSOC arises from both cell types. Finally, FTE- and OSE-derived organoids exhibited differential chemosensitivity. Our results comport with a dualistic origin for HGSOC and suggest that the cell-of-origin might influence therapeutic response.


Treating apraxia of speech with an implicit protocol that activates speech motor areas via inner speech.
Treatments of apraxia of speech (AOS) have traditionally relied on overt practice. One alternative to this method is implicit phoneme manipulation which was derived from early models on inner speech. Implicit phoneme manipulation requires the participant to covertly move and combine phonemes to form a new word. This process engages a system of self-monitoring which is referred to as fully conscious inner speech. The present study aims to advance the understanding and validity of a new treatment for AOS, implicit phoneme manipulation. Tasks were designed to answer the following questions. 1. Would the practice of implicit phoneme manipulation improve the overt production of complex consonant blends in words? 2. Would this improvement generalize to untrained complex and simpler consonant blends in words? 3. Would these treatment tasks activate regions known to support motor planning and programming as verified by fMRI? The participant was asked to covertly manipulate phonemes to create a new word and to associate this newly formed word to a target picture among 4 phonologically-related choices. To avoid overt practice, probes were collected only after each block of training was completed. Probe sessions assessed the effects of implicit practice on the overt production of simple and complex consonant blends in words. An imaging protocol compared semantic baseline tasks to treatment tasks to verify that implicit phoneme manipulation activated brain regions of interest. Behavioral: Response to implicit training of complex consonant blends resulted in improvements which were maintained 6 weeks after treatment. Further, this treatment generalized to simpler consonant blends in words. Imaging: Functional imaging during implicit phoneme manipulation showed significant activation in brain regions responsible for phonological processing when compared to the baseline semantic task. Implicit phoneme manipulation offers an alternative to traditional methods that require overt production for treatment of AOS. Additionally, this implicit treatment method was shown to activate neural areas known to be involved in phonological processing, motor planning and programming.


Chapter 23: Food allergy.
The onset of IgE-mediated food allergy is usually within minutes to 2 hours of food ingestion. Risk factors for fatal food-induced anaphylaxis include presence of asthma (which is a risk factor for anaphylaxis in general), failure to use epinephrine autoinjectors promptly, history of prior severe reactions, known food allergy, denial of symptoms, and adolescent/young adult age. The most commonly implicated foods are cow's milk, eggs, peanuts, soy, tree nuts, fish, shellfish, and wheat. Allergies to peanut, tree nuts, and seafood are the most common food allergens in adults. The major food allergens are glycoproteins that are generally water soluble and stable to the effects of heat, proteases, and acids. Food proteins that escape proteolysis are taken up by intestinal epithelial cells and presented to primed T cells. This process leads to the generation of T-helper type 2 (Th2) cells that produce IL-4, IL-5, and IL-13. Recent studies have found that tolerance can be acquired with >70% of children becoming tolerant to cow's milk and eggs by age 16 years. Allergies to peanuts, tree nuts, and seafood are frequently lifelong. Food-allergic patients or their care givers should be taught when and how to administer injectable epinephrine. In terms of prevention, the American Academy of Pediatrics concluded that there is no convincing evidence that delaying the introduction of solid foods, including common allergens, beyond 4-6 months of age has a protective effect on the development of atopic disease.


Identification and evaluation of epidemic prediction and forecasting reporting guidelines: A systematic review and a call for action.
High quality epidemic forecasting and prediction are critical to support response to local, regional and global infectious disease threats. Other fields of biomedical research use consensus reporting guidelines to ensure standardization and quality of research practice among researchers, and to provide a framework for end-users to interpret the validity of study results. The purpose of this study was to determine whether guidelines exist specifically for epidemic forecast and prediction publications. We undertook a formal systematic review to identify and evaluate any published infectious disease epidemic forecasting and prediction reporting guidelines. This review leveraged a team of 18 investigators from US Government and academic sectors. A literature database search through May 26, 2019, identified 1467 publications (MEDLINE n = 584, EMBASE n = 883), and a grey-literature review identified a further 407 publications, yielding a total 1777 unique publications. A paired-reviewer system screened in 25 potentially eligible publications, of which two were ultimately deemed eligible. A qualitative review of these two published reporting guidelines indicated that neither were specific for epidemic forecasting and prediction, although they described reporting items which may be relevant to epidemic forecasting and prediction studies. This systematic review confirms that no specific guidelines have been published to standardize the reporting of epidemic forecasting and prediction studies. These findings underscore the need to develop such reporting guidelines in order to improve the transparency, quality and implementation of epidemic forecasting and prediction research in operational public health.


Hybrid tandem solar cell enhanced by a metallic hole-array as the intermediate electrode.
A metallic hole-array structure was inserted into a tandem solar cell structure as an intermediate electrode, which allows a further fabrication of a novel and efficient hybrid organic-inorganic tandem solar cell. The inserted hole-array layer reflects the higher-energy photons back to the top cell, and transmits lower-energy photons to the bottom cell via the extraordinary optical transmission (EOT) effect. In this case light absorption in both top and bottom subcells can be simultaneously enhanced via both structural and material optimizations. Importantly, this new design could remove the constraints of requiring lattice-matching and current-matching between the used two cascaded subcells in a conventional tandem cell structure, and therefore, the tunnel junction could be no longer required. As an example, a novel PCBM/CIGS tandem cell was designed and investigated. A systematic modeling study was made on the structural parameter tuning, with the period ranging from a few hundreds nanometers to over one micrometer. Surface plasmon polaritons, magnetic plasmon polaritons, localized surface plasmons, and optical waveguide modes were found to participate in the EOT and the light absorption enhancement. Impressively, more than 40% integrated power enhancement can be achieved in a variable structural parameter range.


Transcriptomics of colour patterning and coloration shifts in crows.
Animal coloration is one of the most conspicuous phenotypic traits in natural populations and has important implications for adaptation and speciation. Changes in coloration can occur over surprisingly short evolutionary timescales, while recurrence of similar colour patterns across large phylogenetic distances is also common. Even though the genetic basis of pigment production is well understood, little is known about the mechanisms regulating colour patterning. In this study, we shed light on the molecular elements regulating regional pigment production in two genetically near-identical crow taxa with striking differences in a eumelanin-based phenotype: black carrion and grey-coated hooded crows. We produced a high-quality genome annotation and analysed transcriptome data from a 2 × 2 design of active melanogenic feather follicles from head (black in both taxa) and torso (black in carrion and grey in hooded crow). Extensive, parallel expression differences between body regions in both taxa, enriched for melanogenesis genes (e.g. ASIP, CORIN, and ALDH6), indicated the presence of cryptic prepatterning also in all-black carrion crows. Meanwhile, colour-specific expression (grey vs. black) was limited to a small number of melanogenesis genes in close association with the central transcription factor MITF (most notably HPGDS, NDP and RASGRF1). We conclude that colour pattern differences between the taxa likely result from an interaction between divergence in upstream elements of the melanogenesis pathway and genes that provide an underlying prepattern across the body through positional information. A model of evolutionary stable prepatterns that can be exposed and masked through simple regulatory changes may explain the phylogenetically independent recurrence of colour patterns that is observed across corvids and many other vertebrate groups.


Multicenter audiometric results with the Vibrant Soundbridge, a semi-implantable hearing device for sensorineural hearing impairment.
The Vibrant Soundbridge, a semi-implantable hearing device for subjects with moderate to severe sensorineural hearing impairment was introduced commercially. First audiologic results are presented on 63 patients from 10 European implant centers. Hearing loss was at 0.5, 1, 2, and 4 kHz varying between 43 and 81 dB HL. The patients used the analogue audio processor, type 302. Measured sound-field gain was compared with NAL-R target values. For most patients an acceptable agreement was found. There was a subgroup of patients, however, with relatively low gain. The results suggest that this was related to the suboptimal positioning and fixation of the transducer to the incus.


Pediatric patient safety in hospitals: a national picture in 2000.
To describe potential patient safety events for hospitalized children, examine associated factors, and explore impacts of safety events. The newly released Patient Safety Indicators (PSIs), developed by researchers at the Agency for Healthcare Research and Quality to identify potential in-hospital patient safety problems using administrative data, were applied to hospital discharge data. All 5.7 million discharge records for children younger than 19 years from 27 states in the 2000 Healthcare Cost and Utilization Project were analyzed for PSI events. Prevalence of PSI events and associations with patient-level and hospital-level characteristics were examined. Multivariate regression adjusting for patient severity of illness was used to estimate impacts of safety events in terms of excess length of stay, charges, and in-hospital mortality. The prevalence of pediatric patient safety events is significant. PSI events occurred more frequently in the very young and those on Medicaid insurance, some of the most vulnerable hospitalized children. Regression analysis found that almost all PSIs are associated with significant and substantial increases in length of stay, charges, and in-hospital death. Using the estimates derived here and the actual number of cases identified in the 2000 data, we estimate that patient safety events incurred >1 billion dollars in excess charges for children alone in 2000. Patient safety problems for hospitalized children occur frequently and with substantial impacts to our health care industry. Unmeasurable by this study are the additional "costs" and "burdens" of safety events that our patients are forced to handle. Additional work to describe and quantify better these outcomes in addition to ones measured here can help solidify the "business case" for patient safety efforts.


Treatment of reticular leg veins with a 1064 nm long-pulsed Nd:YAG laser.
Millisecond-pulsed Nd:YAG lasers have been developed for the treatment of reticular leg veins. We evaluated the effectiveness of a single treatment with a 50 millisecond 1064 nm Nd:YAG laser in the treatment of reticular veins of the lower extremity. Twenty patients with reticular veins measuring 1.0 to 3.0 mm in diameter received one treatment with a 1064 nm Nd:YAG laser (Coolglide, Altus, Burlingame, Calif) at fluences of 100 J/cm(2) and 50 millisecond pulse duration. Symmetric matched areas that were left untreated served as the control. Eleven patients were pretreated with a topical anesthetic cream for 1 hour before treatment and wore compression stockings for 5 days after treatment. Nine patients were treated without topical anesthesia and did not wear compression stockings. Percent clearing and side effects were determined by 3 nontreating physicians (at each respective site) comparing projected Kodachrome images 1 month and 3 months after treatment. Patients also performed a self-assessment of their results. Two-thirds of vessels measuring 1 to 3 mm in diameter cleared more than 75% with one treatment. Larger vessels appeared to improve more than smaller vessels. Immediate treatment discomfort was tolerable. Side effects were minimal and included superficial thrombosis, delayed bruising, hyperpigmentation, and matting. Millisecond-pulsed Nd:YAG lasers used with 50 millisecond pulses are effective in the treatment of reticular leg veins.


Surgical correction of tracheo-oesophageal fistula and oesophageal atresia in infants with VACTERL association: a retrospective case-control study.
VACTERL is a rare, non-random association comprising at least three major component features defined by the acronym, and including Vertebral anomalies, Anorectal malformations, Cardiac defects, Tracheo-oEsophageal fistula with or without oesophageal atresia (TOF/OA), Renal abnormalities and Limb anomalies. The aim of this study was to compare the post-operative outcomes following surgical correction of TOF/OA in infants with VACTERL and isolated TOF/OA. A retrospective case-control study comparing infants with VACTERL (case group) versus infants with isolated TOF/OA (control group) that underwent surgical correction of TOF/OA at our centre between January 2006 and December 2011. Patient demographics, types of anomalies, operative techniques and post-operative outcomes were collected using inpatient and outpatient records. We identified 30 consecutive infants with TOF/OA. Five infants had VACTERL (17 %) and 15 infants had isolated TOF/OA (50 %). There was no significant difference in the gestational age (P = 0.79), birth weight (P = 0.69) or operative repair (P = 0.14) between groups. Overall, surgical correction of TOF/OA led to satisfactory morbidity. Infants with VACTERL were not at higher risk of post-operative complications, such as oesophageal stricture (P = 0.17) or gastro-oesophageal reflux (P = 1.0), compared to infants with isolated TOF/OA. VACTERL association does not increase the risk of post-operative complications following TOF/OA repair.


Integration of the hybrid adenoretroviral vector AdLTR-luc involves both MoMLV elements flanking the transgene.
Vector delivery is still a bottleneck for gene therapy. To overcome some disadvantages of adenoviral and retroviral vectors, we developed a hybrid vector. This hybrid vector, AdLTR-luc, was created by adding two elements from Moloney murine leukemia virus (MoMLV) flanking the luciferase cDNA into an E1/E3-deleted, replication deficient serotype 5 adenovirus vector (Zheng et al., Nature Biotechnol, 2000), and demonstrated that the MoMLV element upstream of the luciferase cDNA was broken during the integration event. The purpose of the current study was to determine if the MoMLV element downstream of the luciferase cDNA was also broken when integration occurred. We used the same A5 cell clones (#10 and 11) from the earlier the paper along with restriction endonuclease digestions, plus Southern hybridization, and PCR. Southern hybridization indicated that the luciferase cDNA was intact in the cloned cells. Results from Xho I and Sal I digestions showed that integration occurred in cloned cells. Southern hybridizations after Nco I digestion suggested that there was a break in both MoMLV elements, upstream and downstream of the luciferase cDNA. After DNA digestion with Not I, hybridization analyses indicated that the MoMLV upstream element was broken during integration. Digestion of genomic DNA with either Xba I/Kpn I, Bam HI/Sac I, or Bam HI/Nco I demonstrated that the MoMLV downstream element was also broken during integration. A PCR assay was unable to amplify the junctional region between the downstream MoMLV element and the adenoviral E2B gene, consistent with a break in that element. Although AdLTR-luc integration is atypical (Zheng et al., Nature Biotechnol, 2000), the present results suggest that both MoMLV elements have important roles in this event.


"I am Primarily Paid for Publishing…": The Narrative Framing of Societal Responsibilities in Academic Life Science Research.
Building on group discussions and interviews with life science researchers in Austria, this paper analyses the narratives that researchers use in describing what they feel responsible for, with a particular focus on how they perceive the societal responsibilities of their research. Our analysis shows that the core narratives used by the life scientists participating in this study continue to be informed by the linear model of innovation. This makes it challenging for more complex innovation models [such as responsible research and innovation (RRI)] to gain ground in how researchers make sense of and conduct their research. Furthermore, the paper shows that the life scientists were not easily able to imagine specific practices that would address broader societal concerns and thus found it hard to integrate the latter into their core responsibilities. Linked to this, researchers saw institutional reward structures (e.g. evaluations, contractual commitments) as strongly focused on scientific excellence ("I am primarily paid for publishing…"). Thus, they saw reward structures as competing with-rather than incentivising-broader notions of societal responsibility. This narrative framing of societal responsibilities is indicative of a structural marginalisation of responsibility practices and explains the claim, made by many researchers in our sample, that they cannot afford to spend time on such practices. The paper thus concludes that the core ideas of RRI stand in tension with predominant narrative and institutional infrastructures that researchers draw on to attribute meaning to their research practices. This suggests that scientific institutions (like universities, professional communities or funding institutions) still have a core role to play in providing new and context-specific narratives as well as new forms of valuing responsibility practices.


Identification of two highly divergent catalase genes in the fungal tomato pathogen, Cladosporium fulvum.
Catalases of pathogenic micro-organisms have attracted attention as potential virulence factors. Homology-based screens were performed to identify catalase genes in the fungal tomato pathogen Cladosporium fulvum. Two highly divergent genes, Cat1 and Cat2, were isolated and characterized. Cat1 codes for a putative 566-amino-acid catalase subunit and belongs to the gene family that also encodes the mainly peroxisome-localized catalases of animal and yeast species. Cat2 codes for a putative catalase subunit of 745 amino acids and belongs to a different gene family coding for the large-subunit catalases similar to ones found in bacteria and filamentous fungi. Neither catalase had an obvious secretory signal sequence. A search for an extracellular catalase was unproductive. The Cat1 and Cat2 genes showed differential expression, with the Cat1 mRNA preferentially accumulating in spores and the Cat2 mRNA preferentially accumulating in response to external H(2)O(2). With Cat2-deleted strains, activity of the Cat2 gene product (CAT2) was identified among four proteins with catalase activity separated on non-denaturing gels. The CAT2 activity represented a minor fraction of the catalase activity in spores and H(2)O(2)-stressed mycelium, and no phenotype was observed for Cat2-deleted strains, which showed a normal response to H(2)O(2) treatment. These results indicate the existence of a complex catalase system in C. fulvum, with regard to both the structure and regulation of the genes involved. In addition, efficient C. fulvum gene-replacement technology has been established.


Local health department food safety and sanitation expenditures and reductions in enteric disease, 2000-2010.
In collaboration with Public Health Practice-Based Research Networks, we investigated relationships between local health department (LHD) food safety and sanitation expenditures and reported enteric disease rates. We combined annual infection rates for the common notifiable enteric diseases with uniquely detailed, LHD-level food safety and sanitation annual expenditure data obtained from Washington and New York state health departments. We used a multivariate panel time-series design to examine ecologic relationships between 2000-2010 local food safety and sanitation expenditures and enteric diseases. Our study population consisted of 72 LHDs (mostly serving county-level jurisdictions) in Washington and New York. While controlling for other factors, we found significant associations between higher LHD food and sanitation spending and a lower incidence of salmonellosis in Washington and a lower incidence of cryptosporidiosis in New York. Local public health expenditures on food and sanitation services are important because of their association with certain health indicators. Our study supports the need for program-specific LHD service-related data to measure the cost, performance, and outcomes of prevention efforts to inform practice and policymaking.