Monitoring aspirin therapy in children after interventional cardiac catheterization: laboratory measures, dose response, and clinical outcomes.
Very few studies have investigated dose response of aspirin and agreement of different platelet function assays in children. One hundred five children were studied at baseline and after interventional cardiac catheterization during aspirin treatment and, in cases of aspirin resistance (AR), after dose increase. Results from arachidonate-induced aggregation (AA) were compared with aggregation induced by ADP, PFA-100 closure times (CTs), urinary 11-dehydro-thromboxane B2 (urinary 11-dhTxB2) levels, and Impact-R % surface coverage. Aspirin at 2-5 mg/kg/day inhibited platelet function in a large majority. While 19 % showed bruising and mild epistaxis, no thrombotic complications were recorded. AR was detected by AA in seven children (6.7 %). After dose increase, the majority showed inhibition by aspirin. Infants had higher urinary 11-dhTxB2 baseline levels; this assay showed some correlation with AA. Both assays manifested high sensitivity and specificity for aspirin while inferior results were found for the other assays. With the PFA-100, 15.2 % of patients were found to have AR, but this corresponded to AR by AA in only one of seven children. While there was poor agreement among assays, AA and urinary 11-dhTxB2 show good specificity for the monitoring of aspirin therapy in children. Aspirin at 2-5 mg/kg inhibits platelet function; AR in children is rare and can be overcome by dose increase.


Lymphedema.
Aggressively applied decongestive measures (eg, manual lymphatic drainage, low-stretch bandaging, exercise, skin care, application of compressive elastic garments) are the mainstay of lymphatic therapy. Therapeutic regimens should differentiate between the goals of acute volume reduction and the maintenance of limb volume. Elastic garments should not be employed until maximal volume reduction has been attained through decongestive lymphatic techniques. It is my opinion that use of intermittent pneumatic compression devices can play an important adjunctive role to decongestive lymphatic therapy but should not be substituted for these techniques. At this time, I am not inclined to use pharmacologic therapy in these patients but anxiously await the results of studies that might demonstrate efficacy for molecular approaches. Surgical intervention is reserved for a small number of well-selected patients. Liposuction for volume reduction appears to be a very promising approach for specific patients.


APeg3: regulation of Peg3 through an evolutionarily conserved ncRNA.
Mammalian APeg3 is an antisense gene that is localized within the 3'-untranslated region of the imprinted gene, Peg3. APeg3 is expressed only in the vasopressinergic neurons of the hypothalamus, thus is predicted to play significant roles in this specific area of the brain. In the current study, we investigate the functions of APeg3 with comparative genomics and cell line-based functional approaches. The transcribed region of APeg3 displays high levels of sequence conservation among placental mammals, but without any obvious open reading frame, suggesting that APeg3 may have been selected as a ncRNA gene during eutherian evolution. This has been further supported by the detection of a conserved local RNA secondary structure within APeg3. RNA secondary structure analyses indicate a single conserved hairpin-loop structure towards the 5' end of the transcript. The results from cell line-based transfection experiments demonstrate that APeg3 has the potential to down-regulate the transcription and protein levels of Peg3. The observed down-regulation by APeg3 is also somewhat orientation-independent. Overall, these results suggest that APeg3 has evolved as a ncRNA gene and controls the function of its sense gene Peg3 within specific neuronal cells.


Automatic change detection of driving environments in a vision-based driver assistance system.
Detecting critical changes of environments while driving is an important task in driver assistance systems. In this paper, a computational model motivated by human cognitive processing and selective attention is proposed for this purpose. The computational model consists of three major components, referred to as the sensory, perceptual, and conceptual analyzers. The sensory analyzer extracts temporal and spatial information from video sequences. The extracted information serves as the input stimuli to a spatiotemporal attention (STA) neural network embedded in the perceptual analyzer. If consistent stimuli repeatedly innervate the neural network, a focus of attention will be established in the network. The attention pattern associated with the focus, together with the location and direction of motion of the pattern, form what we call a categorical feature. Based on this feature, the class of the attention pattern and, in turn, the change in driving environment corresponding to the class are determined using a configurable adaptive resonance theory (CART) neural network, which is placed in the conceptual analyzer. Various changes in driving environment, both in daytime and at night, have been tested. The experimental results demonstrated the feasibilities of both the proposed computational model and the change detection system.


Routine habitat change: a source of unrecognized transient alteration of intestinal microbiota in laboratory mice.
The mammalian intestine harbors a vast, complex and dynamic microbial population, which has profound effects on host nutrition, intestinal function and immune response, as well as influence on physiology outside of the alimentary tract. Imbalance in the composition of the dense colonizing bacterial population can increase susceptibility to various acute and chronic diseases. Valuable insights on the association of the microbiota with disease critically depend on investigation of mouse models. Like in humans, the microbial community in the mouse intestine is relatively stable and resilient, yet can be influenced by environmental factors. An often-overlooked variable in research is basic animal husbandry, which can potentially alter mouse physiology and experimental outcomes. This study examined the effects of common husbandry practices, including food and bedding alterations, as well as facility and cage changes, on the gut microbiota over a short time course of five days using three culture-independent techniques, quantitative PCR, terminal restriction fragment length polymorphism (TRFLP) and next generation sequencing (NGS). This study detected a substantial transient alteration in microbiota after the common practice of a short cross-campus facility transfer, but found no comparable alterations in microbiota within 5 days of switches in common laboratory food or bedding, or following an isolated cage change in mice acclimated to their housing facility. Our results highlight the importance of an acclimation period following even simple transfer of mice between campus facilities, and highlights that occult changes in microbiota should be considered when imposing husbandry variables on laboratory animals.


First clinical experiences with a new ovine Fab Echis ocellatus snake bite antivenom in Nigeria: randomized comparative trial with Institute Pasteur Serum (Ipser) Africa antivenom.
During the past decade, effective snake antivenoms have become scarce in northern Nigeria. As a result, many patients severely envenomed by the saw-scaled or carpet viper (Echis ocellatus), which is responsible for more than 95% of the snake bites in the region, did not receive effective treatment and mortality and morbidity increased. To combat this crisis, a new monospecific ovine Fab antivenom (EchiTab) is being developed. Its theoretical advantages over conventional equine F(ab')2 antivenom are a more rapid tissue penetration and larger apparent volume of distribution (the volume of [tissue] fluid in which the the antivenom would be uniformly distributed to achieve the observed plasma concentration). In a preliminary study, two vials (20 ml; 1.0 g of protein) of EchiTab rapidly and permanently restored blood coagulability and cleared venom antigenemia in seven envenomed patients. Four experienced early reactions that responded to epinephrine. In a randomized comparative trial of one vial (10 ml; 0.5 g protein) of EchiTab or four ampules (40 ml; 2.12 g of protein) of Institute Pasteur Serum (Ipser) Africa polyspecific F(ab')2 antivenom, there were fewer reactions, but only 36% and 35% of patients, respectively, showed permanent restoration of coagulability, with the remainder requiring further doses. This suggests that 0.5 g (one vial) of EchiTab is approximately equivalent to 2.12 g (four ampules) of Ipser Africa antivenom, and that a higher initial dose will be required for most patients. Measurements of circulating venom and antivenom levels reflected the clinical events.


Investigation of bacterial diversity in the feces of cattle fed different diets.
The objective of this study is to investigate individual animal variation of bovine fecal microbiota including as affected by diets. Fecal samples were collected from 426 cattle fed 1 of 3 diets typically fed to feedlot cattle: 1) 143 steers fed finishing diet (83% dry-rolled corn, 13% corn silage, and 4% supplement), 2) 147 steers fed late growing diet (66% dry-rolled corn, 26% corn silage, and 8% supplement), and 3) 136 heifers fed early growing diet (70% corn silage and 30% alfalfa haylage). Bacterial 16S rRNA gene amplicons were determined from individual fecal samples using next-generation pyrosequencing technology. A total of 2,149,008 16S rRNA gene sequences from 333 cattle with at least 2,000 sequences were analyzed. Firmicutes and Bacteroidetes were dominant phyla in all fecal samples. At the genus level, Oscillibacter, Turicibacter, Roseburia, Fecalibacterium, Coprococcus, Clostridium, Prevotella, and Succinivibrio were represented by more than 1% of total sequences. However, numerous sequences could not be assigned to a known genus. Dominant unclassified groups were unclassified Ruminococcaceae and unclassified Lachnospiraceae that could be classified to a family but not to a genus. These dominant genera and unclassified groups differed (P < 0.001) with diets. A total of 176,692 operational taxonomic units (OTU) were identified in combination across all the 333 cattle. Only 2,359 OTU were shared across 3 diet groups. UniFrac analysis showed that bacterial communities in cattle feces were greatly affected by dietary differences. This study indicates that the community structure of fecal microbiota in cattle is greatly affected by diet, particularly between forage- and concentrate-based diets.


Substance use and young people: the potential of technology.
There is widespread concern about alcohol and drug use and abuse among young people in the United Kingdom, Europe and the United States. Evaluations of current drug and alcohol education approaches have mixed findings and some methodological difficulties. This paper reports on exploratory research to explore the potential of technology to provide information and support to young people regarding substance use and abuse. Eight focus groups were conducted with young people (n = 78) and three with key informants (n = 22). The findings revealed that technology has potential to provide information on this topic, and that young people are critical of some of the more traditional methods of provision. The young people were prepared to experiment with technology, felt competent to do so, and made suggestions of information they would like. These findings are discussed and recommendations made for future research in the area.


Molecular characterization of eukaryotic algal communities in the tropical phyllosphere based on real-time sequencing of the 18S rDNA gene.
Foliicolous algae are a common occurrence in tropical forests. They are referable to a few simple morphotypes (unicellular, sarcinoid-like or filamentous), which makes their morphology of limited usefulness for taxonomic studies and species diversity assessments. The relationship between algal community and their host phyllosphere was not clear. In order to obtain a more accurate assessment, we used single molecule real-time sequencing of the 18S rDNA gene to characterize the eukaryotic algal community in an area of South-western China. We annotated 2922 OTUs belonging to five classes, Ulvophyceae, Trebouxiophyceae, Chlorophyceae, Dinophyceae and Eustigmatophyceae. Novel clades formed by large numbers sequences of green algae were detected in the order Trentepohliales (Ulvophyceae) and the Watanabea clade (Trebouxiophyceae), suggesting that these foliicolous communities may be substantially more diverse than so far appreciated and require further research. Species in Trentepohliales, Watanabea clade and Apatococcus clade were detected as the core members in the phyllosphere community studied. Communities from different host trees and sampling sites were not significantly different in terms of OTUs composition. However, the communities of Musa and Ravenala differed from other host plants significantly at the genus level, since they were dominated by Trebouxiophycean epiphytes. The cryptic diversity of eukaryotic algae especially Chlorophytes in tropical phyllosphere is very high. The community structure at species-level has no significant relationship either with host phyllosphere or locations. The core algal community in tropical phyllopshere is consisted of members from Trentepohliales, Watanabea clade and Apatococcus clade. Our study provided a large amount of novel 18S rDNA sequences that will be useful to unravel the cryptic diversity of phyllosphere eukaryotic algae and for comparisons with similar future studies on this type of communities.


The Arabidopsis RNA-binding protein AtRGGA regulates tolerance to salt and drought stress.
Salt and drought stress severely reduce plant growth and crop productivity worldwide. The identification of genes underlying stress response and tolerance is the subject of intense research in plant biology. Through microarray analyses, we previously identified in potato (Solanum tuberosum) StRGGA, coding for an Arginine Glycine Glycine (RGG) box-containing RNA-binding protein, whose expression was specifically induced in potato cell cultures gradually exposed to osmotic stress. Here, we show that the Arabidopsis (Arabidopsis thaliana) ortholog, AtRGGA, is a functional RNA-binding protein required for a proper response to osmotic stress. AtRGGA gene expression was up-regulated in seedlings after long-term exposure to abscisic acid (ABA) and polyethylene glycol, while treatments with NaCl resulted in AtRGGA down-regulation. AtRGGA promoter analysis showed activity in several tissues, including stomata, the organs controlling transpiration. Fusion of AtRGGA with yellow fluorescent protein indicated that AtRGGA is localized in the cytoplasm and the cytoplasmic perinuclear region. In addition, the rgga knockout mutant was hypersensitive to ABA in root growth and survival tests and to salt stress during germination and at the vegetative stage. AtRGGA-overexpressing plants showed higher tolerance to ABA and salt stress on plates and in soil, accumulating lower levels of proline when exposed to drought stress. Finally, a global analysis of gene expression revealed extensive alterations in the transcriptome under salt stress, including several genes such as ASCORBATE PEROXIDASE2, GLUTATHIONE S-TRANSFERASE TAU9, and several SMALL AUXIN UPREGULATED RNA-like genes showing opposite expression behavior in transgenic and knockout plants. Taken together, our results reveal an important role of AtRGGA in the mechanisms of plant response and adaptation to stress.


Molecular analysis demonstrates high prevalence of chloroquine resistance but no evidence of artemisinin resistance in Plasmodium falciparum in the Chittagong Hill Tracts of Bangladesh.
Artemisinin resistance is present in the Greater Mekong region and poses a significant threat for current anti-malarial treatment guidelines in Bangladesh. The aim of this molecular study was to assess the current status of drug resistance in the Chittagong Hill Tracts of Bangladesh near the Myanmar border. Samples were obtained from patients enrolled into a Clinical Trial (NCT02389374) conducted in Alikadam, Bandarban between August 2014 and January 2015. Plasmodium falciparum infections were confirmed by PCR and all P. falciparum positive isolates genotyped for the pfcrt K76T and pfmdr1 N86Y markers. The propeller region of the kelch 13 (k13) gene was sequenced from isolates from patients with delayed parasite clearance. In total, 130 P. falciparum isolates were available for analysis. The pfcrt mutation K76T, associated with chloroquine resistance was found in 81.5% (106/130) of cases and the pfmdr1 mutation N86Y in 13.9% (18/130) cases. No single nucleotide polymorphisms were observed in the k13 propeller region. This study provides molecular evidence for the ongoing presence of chloroquine resistant P. falciparum in Bangladesh, but no evidence of mutations in the k13 propeller domain associated with artemisinin resistance. Monitoring for artemisinin susceptibility in Bangladesh is needed to ensure early detection and containment emerging anti-malarial resistance.


Recognition of DHN-melanin by a C-type lectin receptor is required for immunity to Aspergillus.
Resistance to infection is critically dependent on the ability of pattern recognition receptors to recognize microbial invasion and induce protective immune responses. One such family of receptors are the C-type lectins, which are central to antifungal immunity. These receptors activate key effector mechanisms upon recognition of conserved fungal cell-wall carbohydrates. However, several other immunologically active fungal ligands have been described; these include melanin, for which the mechanism of recognition is hitherto undefined. Here we identify a C-type lectin receptor, melanin-sensing C-type lectin receptor (MelLec), that has an essential role in antifungal immunity through recognition of the naphthalene-diol unit of 1,8-dihydroxynaphthalene (DHN)-melanin. MelLec recognizes melanin in conidial spores of Aspergillus fumigatus as well as in other DHN-melanized fungi. MelLec is ubiquitously expressed by CD31+ endothelial cells in mice, and is also expressed by a sub-population of these cells that co-express epithelial cell adhesion molecule and are detected only in the lung and the liver. In mouse models, MelLec was required for protection against disseminated infection with A. fumigatus. In humans, MelLec is also expressed by myeloid cells, and we identified a single nucleotide polymorphism of this receptor that negatively affected myeloid inflammatory responses and significantly increased the susceptibility of stem-cell transplant recipients to disseminated Aspergillus infections. MelLec therefore recognizes an immunologically active component commonly found on fungi and has an essential role in protective antifungal immunity in both mice and humans.


Stable sub-85 fs passively mode-locked Erbiumfiber oscillator with tunable repetition rate.
We present a sub-85 fs self-starting stretched-pulse passively mode-locked Erbium-fiber oscillator in a sigma setup with tunable repetition rate. The sigma cavity included a movable mirror enabling a tunable pulse repetition rate variation of +/- 1 % from 55.3 MHz to 56.4 MHz with continuous, uninterrupted mode-locked operation and an output power around 14 mW. Based on the wide tuning range of the repetition rate the presented fiber oscillator is a suitable candidate for applications in femtosecond spectroscopy or precision metrology around 1.56 microm.


Structural studies of the effect that dimethyl sulfoxide (DMSO) has on cisplatin and carboplatin binding to histidine in a protein.
The anticancer complexes cisplatin and carboplatin target the DNA major groove, forming intrastrand and interstrand cross-links between guanine bases through their N7 atoms, causing distortion of the DNA helix and apoptotic cell death. A major side effect of these drugs is toxicity, which is caused via binding to many proteins in the body. A range of crystallographic studies have been carried out involving the cocrystallization of hen egg-white lysozyme (HEWL) as a test protein with cisplatin and carboplatin in aqueous and dimethyl sulfoxide (DMSO) conditions. Different cryoprotectants, glycerol and Paratone, were used for each of the cisplatin and carboplatin cocrystallization cases, while silicone oil was used for studies involving N-acetylglucosamine (NAG). Both cisplatin and carboplatin do not bind to HEWL in aqueous media on the timescales of the conditions used here, but upon addition of DMSO two molecules of cisplatin or carboplatin bind either side of His15, which is the only His residue in lysozyme and is assumed to be an imidazolyl anion or a chemical resonance moiety, i.e. both imidazole N atoms are chemically reactive. To identify the platinum-peak positions in the 'with DMSO conditions', anomalous scattering maps were calculated as a cross-check with the F(o) - F(c) OMIT maps. Platinum-occupancy σ values were established using three different software programs in each case. The use of EVAL15 to process all of the diffraction data sets provided a consistent platform for a large ensemble of data sets for the various protein and platinum-compound model refinements with REFMAC5 and then SHELXTL. Overall, this extensive set of crystallization and cryoprotectant conditions allowed a systematic evaluation of cisplatin and carboplatin binding to lysozyme as a test protein via detailed X-ray crystal structure characterizations. DMSO is used as a super-solvent for drug delivery as it is deemed to cause no effect upon drug binding. However, these results show that addition of DMSO causes the platinum anticancer drugs to bind to HEWL. This effect should be considered in toxicity assessments of these drugs and perhaps more widely.


[Formula: see text]Official Position of the American Academy of Clinical Neuropsychology (AACN): Guidelines for Practicum Training in Clinical Neuropsychology.
Practical experience is central to the education and training of neuropsychologists, beginning in graduate school and extending through postdoctoral fellowship. However, historically, little attention has been given to the structure and requirements of practicum training in clinical neuropsychology. A working group of senior-level neuropsychologists, as well as a current postdoctoral fellow, all from a diverse range of settings (The AACN Practicum Guidelines Workgroup), was formed to propose guidelines for practicum training in clinical neuropsychology. The Workgroup reviewed relevant literature and sought input from professional organizations involved in education and training in neuropsychology. The proposed guidelines provide a definition of practicum training in clinical neuropsychology, detail entry and exit criteria across competencies relevant to practicum training in clinical neuropsychology, and discuss the relationship between doctoral training programs and practicum training sites. The proposed guidelines also provide a methodology for competency-based evaluation of clinical neuropsychology practicum trainees and outline characteristics and features that are integral to an effective training environment. Although the guidelines discussed below may not be implemented in their entirety across all clinical neuropsychology practicum training sites, they are consistent with the latest developments in competency-based education.


Does light influence the relationship between a native stem hemiparasite and a native or introduced host?
There have been very few studies investigating the influence of light on the effects of hemiparasitic plants on their hosts, despite the fact that hemiparasites are capable of photosynthesis but also access carbon (C) from their host. In this study we manipulated light availability to limit photosynthesis in an established hemiparasite and its hosts, and determined whether this affected the parasite's impact on growth and performance of two different hosts. We expected that limiting light and reducing autotrophic C gain in the parasite (and possibly increasing its heterotrophic C gain) would lead to an increased impact on host growth and/or host photosynthesis in plants grown in low (LL) relative to high light (HL). The Australian native host Leptospermum myrsinoides and the introduced host Ulex europaeus were either infected or not infected with the native stem hemiparasite Cassytha pubescens and grown in either HL or LL. Photosynthetic performance, nitrogen status and growth of hosts and parasite were quantified. Host water potentials were also measured. In situ midday electron transport rates (ETRs) of C. pubescens on both hosts were significantly lower in LL compared with HL, enabling us to investigate the impact of the reduced level of parasite autotrophy on growth of hosts. Despite the lower levels of photosynthesis in the parasite, the relative impact of infection on host biomass was the same in both LL and HL. In fact, biomass of L. myrsinoides was unaffected by infection in either HL or LL, while biomass of U. europaeus was negatively affected by infection in both treatments. This suggests that although photosynthesis of the parasite was lower in LL, there was no additional impact on host biomass in LL. In addition, light did not affect the amount of parasite biomass supported per unit host biomass in either host, although this parameter was slightly lower in LL than HL for U. europaeus (P = 0·073). We also found no significant enhancement of host photosynthesis in response to infection in either host, regardless of light treatment. Despite lower photosynthetic rates in LL, C. pubescens did not increase its dependency on host C to the point where it affected host growth or photosynthesis. The impact of C. pubescens on host growth would be similar in areas of high and low light availability in the field, but the introduced host is more negatively affected by infection.


Divergent habitat use of two urban lizard species.
Faunal responses to anthropogenic habitat modification represent an important aspect of global change. In Puerto Rico, two species of arboreal lizard, Anolis cristatellus and A. stratulus, are commonly encountered in urban areas, yet seem to use the urban habitat in different ways. In this study, we quantified differences in habitat use between these two species in an urban setting. For each species, we measured habitat use and preference, and the niche space of each taxon, with respect to manmade features of the urban environment. To measure niche space of these species in an urban environment, we collected data from a total of six urban sites across four different municipalities on the island of Puerto Rico. We quantified relative abundance of both species, their habitat use, and the available habitat in the environment to measure both microhabitat preference in an urban setting, as well as niche partitioning between the two different lizards. Overall, we found that the two species utilize different portions of the urban habitat. Anolis stratulus tends to use more "natural" portions of the urban environment (i.e., trees and other cultivated vegetation), whereas A. cristatellus more frequently uses anthropogenic structures. We also found that aspects of habitat discrimination in urban areas mirror a pattern measured in prior studies for forested sites in which A. stratulus was found to perch higher than A. cristatellus and preferred lower temperatures and greater canopy cover. In our study, we found that the multivariate niche space occupied by A. stratulus did not differ from the available niche space in natural portions of the urban environment and in turn represented a subset of the niche space occupied by A. cristatellus. The unique niche space occupied by A. cristatellus corresponds to manmade aspects of the urban environment generally not utilized by A. stratulus. Our results demonstrate that some species are merely tolerant of urbanization while others utilize urban habitats in novel ways. This finding has implications for long-term persistence in urban habitats and suggests that loss of natural habitat elements may lead to nonrandom species extirpations as urbanization intensifies.


Regulatory hotspots are associated with plant gene expression under varying soil phosphorus supply in Brassica rapa.
Gene expression is a quantitative trait that can be mapped genetically in structured populations to identify expression quantitative trait loci (eQTL). Genes and regulatory networks underlying complex traits can subsequently be inferred. Using a recently released genome sequence, we have defined cis- and trans-eQTL and their environmental response to low phosphorus (P) availability within a complex plant genome and found hotspots of trans-eQTL within the genome. Interval mapping, using P supply as a covariate, revealed 18,876 eQTL. trans-eQTL hotspots occurred on chromosomes A06 and A01 within Brassica rapa; these were enriched with P metabolism-related Gene Ontology terms (A06) as well as chloroplast- and photosynthesis-related terms (A01). We have also attributed heritability components to measures of gene expression across environments, allowing the identification of novel gene expression markers and gene expression changes associated with low P availability. Informative gene expression markers were used to map eQTL and P use efficiency-related QTL. Genes responsive to P supply had large environmental and heritable variance components. Regulatory loci and genes associated with P use efficiency identified through eQTL analysis are potential targets for further characterization and may have potential for crop improvement.


Idiopathic intracranial hypertension.
Idiopathic intracranial hypertension (IIH) is a condition which affects predominantly overweight women and is characterized by raised intracranial pressure without any identifiable pathology in the brain and with normal cerebrospinal fluid (CSF) composition. The cause of IIH is unclear and as such it remains a diagnosis of exclusion. Although the pathophysiology of IIH remains elusive, some observations have recently been added to our understanding of this, including the presence of transverse sinus stenosis on many patients and the possible role of leptin and inflammation in the disease pathogenesis. Headache is the most common symptom and papilloedema is the major clinical finding. Choices of medical treatment are limited to the use of diuretics particularly acetazolamide and encouragement of weight loss. Surgical therapies such as CSF diversion procedures and fenestration of the optic nerve may be necessary in some cases with persistent symptoms or progressive visual deterioration. While not life-threatening, IIH has a significant morbidity with up to 25% of patients developing visual impairment from optic atrophy. Visual surveillance is therefore vital. Long-term follow-up is recommended as the disease may worsen after an initial period of stability.


A comprehensive approach providing a new synthetic route for bimetallic electrocatalysts via isoPOMs [M/Rh(Cp*)4W8O32] (M = Rh (1) and Ir (2)).
Polyoxometalates (POMs) are highly versatile materials if paired with various conducting supports. We here present a new prospective approach for obtaining advanced electrocatalytic systems via POMs (1 and 2) nanostructured on Ni foam (NF) to give bimetallic nanohybrid composite materials M0.4Ni0.6/WO3 (M = Rh (3) or Ir (4)). By this pathway, two rare isopolyoctatungstate-based organometallic clusters [Hx(MCp*)4W8O32]·yH2O (in 1, M = Rh, x = 8, y = 23; in 2, M = Ir, x = 4, y = 17.33) have been synthesized via one-pot reactions using [Cp*MCl2]2 (Cp* = pentamethylcyclopentadienyl, M = Rh or Ir) and Na2WO4·2H2O in basic conditions. Compounds 1 and 2 were successfully nanostructured on the surface of commercially available NF and were thermally reduced to M0.4Ni0.6/WO3 (M = Rh (3) or Ir (4)) after annealing at 400 °C for 8 h in a 15% H2/N2 atmosphere. Electrodes modified with the POMs 3 and 4 exhibited remarkable HER activities in a 1 M KOH solution and achieved a current density of 10 mA cm-2 at low overpotentials of about 67 and 35 mV versus RHE, with Tafel slopes of 56 and 34 mV dec-1, respectively.


Multicolor mapping of the cardiomyocyte proliferation dynamics that construct the atrium.
The orchestrated division of cardiomyocytes assembles heart chambers of distinct morphology. To understand the structural divergence of the cardiac chambers, we determined the contributions of individual embryonic cardiomyocytes to the atrium in zebrafish by multicolor fate-mapping and we compare our analysis to the established proliferation dynamics of ventricular cardiomyocytes. We find that most atrial cardiomyocytes become rod-shaped in the second week of life, generating a single-muscle-cell-thick myocardial wall with a striking webbed morphology. Inner pectinate myofibers form mainly by direct branching, unlike delamination events that create ventricular trabeculae. Thus, muscle clones assembling the atrial chamber can extend from wall to lumen. As zebrafish mature, atrial wall cardiomyocytes proliferate laterally to generate cohesive patches of diverse shapes and sizes, frequently with dominant clones that comprise 20-30% of the wall area. A subpopulation of cardiomyocytes that transiently express atrial myosin heavy chain (amhc) contributes substantially to specific regions of the ventricle, suggesting an unappreciated level of plasticity during chamber formation. Our findings reveal proliferation dynamics and fate decisions of cardiomyocytes that produce the distinct architecture of the atrium.


The Development of Theory of Mind and Positive and Negative Reciprocity in Preschool Children.
This study examined the relation between the acquisition of false-beliefs theory of mind (ToM) and reciprocity in preschoolers. Preschool-aged children completed a task assessing the understanding of false beliefs, and played an Ultimatum Game (UG) with another child in a face-to-face setting. Negative reciprocity was assessed by examining the rejection of unfair offers made by another child in the UG, while positive reciprocity was assessed by examining allocations made by participants in a Dictator Game (DG) following the UG. The results indicated that children who had passed a task assessing first-order false beliefs were more likely to make generous offers in a DG following a fair offer made by their partner in a proceeding UG, but that false beliefs ToM was unrelated to the rejection of unfair offers in the UG.


An Intrathermocline Eddy and a tropical cyclone in the Bay of Bengal.
The Bay of Bengal, subjected to monsoonal forcing and tropical cyclones, displays a complex field of ocean eddies. On 5 December 2013 a sub-surface vortex or Intrathermocline Eddy (ITE) composed of water characteristic of the Andaman Sea was observed within the thermocline of the western Bay of Bengal. We propose that the ITE was the product of Tropical Cyclone Lehar interaction on 27 November 2013 with a westward propagating surface eddy from the eastern Bay of Bengal. While Lehar's interaction with the ocean initially removes heat from the upper layers of the eddy, air-sea flux is limited as the deeper portions of the eddy was subducted into the stratified thermocline, inhibiting further interaction with the atmosphere. The ITE core from 30 to 150 m is thus isolated from local air-sea fluxes by strong stratification at the mixed layer base, and its periphery is stable to shear instability, suggestive of longevity and the ability to carry water far distances with minimal modification.


Estrogen-induced increase in the magnitude of long-term potentiation occurs only when the ratio of NMDA transmission to AMPA transmission is increased.
Elevated levels of estradiol enhance learning in mammals, including humans, likely a result of hormone-induced heightened plasticity at CA3-CA1 synapses. The increase in long-term potentiation (LTP) magnitude is considered to be a consequence of the estradiol-induced increase in dendritic spine density and NMDA receptor (NMDAR)-mediated transmission; however, direct evidence linking these changes together is lacking. Alternatively, alterations in GABAergic inhibition or presynaptic release probability could contribute. Here, we show in time course studies using hippocampal slices from estradiol-treated ovariectomized rats that the LTP magnitude is increased only when spine density is increased simultaneously with an increase in NMDAR transmission relative to AMPA receptor (AMPAR) transmission, with no role for alterations in GABAergic inhibition or release probability. With time after hormone treatment, AMPAR transmission gradually increases during the maintained increase in spine density and NMDAR transmission. Eventually, the balance between NMDAR and AMPAR transmission is reestablished, and the LTP magnitude is no longer increased. Blocking genomic estrogen receptors prevents the heightened spine density, NMDAR transmission, and LTP magnitude, suggesting a tight mechanistic coupling between these morphological and functional changes. Thus, we propose that the hormone-induced increase in functional synapse density alone is not sufficient to support heightened plasticity. Rather, estradiol increases LTP via enhancing NMDAR transmission, likely through receptor insertion into newly formed or preexisting synapses. Later, when excitability in the circuit is at its highest and spine density remains elevated, the LTP magnitude is no longer increased, probably as a consequence of the delayed increase in AMPAR transmission that resets the balance between NMDAR and AMPAR transmission.


Rapid Characterization of Emerging Per- and Polyfluoroalkyl Substances in Aqueous Film-Forming Foams Using Ion Mobility Spectrometry-Mass Spectrometry.
Aqueous film-forming foams (AFFF) are mixtures formulated with numerous hydrocarbon- and fluoro-containing surfactants. AFFF use leads to environmental releases of unknown per- and polyfluoroalkyl substances (PFAS). AFFF composition is seldom disclosed, and their use elicits concerns from both regulatory agencies and the public because PFAS are persistent in the environment and potentially associated with adverse health effects. In this study, we demonstrate the use of coupled liquid chromatography, ion mobility spectrometry, and mass spectrometry (LC-IMS-MS) to rapidly characterize both known and unknown PFAS in AFFF. Ten AFFF formulations from seven brands were analyzed using LC-IMS-MS in both negative and positive ion modes. Untargeted analysis of the formulations was followed by feature identification of PFAS-like features utilizing database matching, mass defect and homologous series evaluation, and MS/MS fragmentation experiments. Across the tested AFFF formulations, we identified 33 homologous series; only ten of these homologous series have been previously reported. Among tested AFFF, the FireStopper (n = 85) contained the greatest number of PFAS-like features and Phos-Check contained zero. This work demonstrates that LC-IMS-MS-enabled untargeted analysis of complex formulations, followed by feature identification using data-processing algorithms, can be used for rapid exposure characterization of known and putative PFAS during fire suppression-related contamination events.


Complete Genome Sequences of Five Salmonella enterica Strains Used in Inoculation Cocktails in Low-Moisture Food Storage Studies.
Survival kinetics of Salmonella enterica have been previously studied using an inoculum cocktail composed of different strains that have been associated with low-moisture foods. Here, we report the closed genome sequences of five strains of Salmonella enterica that are commonly used in these storage studies.


Diurnal variation in the renal excretion of modified RNA catabolites in humans.
Renally excreted modified RNA catabolites [pseudouridine (psi), N (2), N (2)-dimethylguanosine (m(2)(2)G) and N (6)-threoninocarbonyladenosine (t(6)A)] are markers of whole-body rates of degradation of rRNA and tRNA, and are thought to be potential indicators of the resting metabolic rate. To investigate diurnal variations of these RNA catabolites, the amounts of psi, m(2)(2)G and t(6)A excreted were determined by HPLC of the urine from eight healthy male adults collected over 47-h periods, which were subdivided into the morning (06.00 or 09.00 to 12.00 hours), the afternoon (12.00 to 18.00 hours), the evening (18.00 to 24.00 hours) and the night (00.00 to 06.00 or 08.00 hours), under two different nutritional regimens with 100 or 50 g of protein/day. Furthermore, rates of degradation of rRNA and tRNA were calculated using values for these RNA catabolites. For comparison, the corresponding excretion of creatinine, which originates from the energy metabolism of muscle, and of 3-methylhistidine (m(3)His), which is an indicator of muscle protein degradation, was determined. Differences in excretion during the collection periods were tested using the Friedmann test. The excretion of psi, creatinine and m(3)His (micromol x h(-1) x kg(-1)) altered significantly (P <0.001) during the day. Medians were: for psi, 0.21 (morning), 0.19 (afternoon), 0.19 (evening) and 0.18 (night); for creatinine, 8.8, 8.4, 8.0 and 7.3 respectively; for m(3)His, 0.13, 0.11, 0.12 and 0.10 respectively. The excretion rates of m(2)(2)G and t(6)A (nmol x h(-1) x kg(-1)) altered, but not significantly, during the day; corresponding medians were: for m(2)(2)G, 9.0, 8.4, 8.0 and 8.4 respectively; for t(6)A, 4.3, 4.1, 3.9 and 3.9 respectively. From these results it can be concluded that, in order to assess the average daily rates of degradation of tRNA and rRNA using modified RNA catabolites, urine collection should be carried out quantitatively over 24 h periods. Likewise, the catabolites creatinine and m(3)His must be determined using 24 h urine samples when average daily excretion values are required.


Application of the screening method to monitor influenza vaccine effectiveness among the elderly in Germany.
Elderly people are at increased risk for severe influenza illness and constitute therefore a major target-group for seasonal influenza vaccination in most industrialized countries. The aim of this study was to estimate influenza vaccine effectiveness (VE) among individuals aged 60+ years over three seasons and to assess if the screening method is a suitable tool to monitor influenza VE in this particular target-group in Germany. We identified laboratory-confirmed influenza cases aged 60+ years through the national communicable disease reporting system for seasons 2010/11, 2011/12 and 2012/13. Vaccination coverage (VC) data were retrieved from a database of health insurance claims representing ~85% of the total German population. We applied the screening method to calculate influenza subtype-specific VE and compared our results with VE estimates from other observational studies in Europe. In total, 7,156 laboratory-confirmed influenza cases were included. VE against all influenza types ranged between 49% (95% confidence interval [CI]: 39-56) in 2011/12 and 80% (95% CI: 76-83%) in 2010/11. In 2010/11 subtype-specific VE against influenza A(H1N1)pdm and B was 76% and 84%, respectively. In the following seasons, VE against influenza A(H1N1)pdm, A(H3N2) and B was 87%, -9% , 74% (2011/12), and 74%, 39%, 73% (2012/13). VE was higher among hospitalized compared to non-hospitalized influenza A cases. Seventeen observational studies from Europe reporting subtype-specific VE among the elderly were identified for the respective seasons (all applying the test-negative design) and showed comparable subtype-specific VE estimates. According to our study, influenza vaccination provided moderate protection against laboratory-confirmed influenza A(H1N1)pdm and B in individuals aged 60+ but no or only little protection against A(H3N2). Higher VE among hospitalized cases might indicate higher protection against severe influenza disease. Based on the available data, the screening method allowed us to assess subtype-specific VE in hospitalized and non-hospitalized elderly persons. Since controlling for several important confounders was not possible, the applied method only provided crude VE estimates. However, given the precise VC-data and the large number of cases, the screening method provided results being in line with VE estimates from other observational studies in Europe that applied a different study design.


Conserved mammalian gonadotropin-releasing hormone receptor carboxyl terminal amino acids regulate ligand binding, effector coupling and internalization.
The mammalian gonadotropin-releasing hormone receptor (GnRHR), with 327 amino acids, is among the smallest G protein coupled receptors identified. Absent from this receptor is the cytoplasmic tail, characteristic of other members of this superfamily, which frequently mediates desensitization and down-regulation. The fifteen carboxyl terminal residues in the mammalian GnRHR are absolutely conserved, suggesting important roles for these residues. In the current study, mutations of the mammalian GnRHR were made to study the carboxyl terminus. The receptor mutant GnRHR(Ser(326)Ala) was reduced in ligand affinity (117% reduction compared to wild type (wt)), while receptor numbers and internalization remained unchanged. GnRHR(Ser(326)Tyr) was decreased in effector coupling, while ligand affinity remained unchanged compared to wt. These studies also show that, while mutation of Ser(326) caused a change in ligand binding and effector coupling, truncation at this residue (GnRHR[des(326-327)]) had no measurable effect on GnRHR ligand binding, effector coupling or internalization, functions which appear to require different structural determinants than expression and routing. Removal of all three carboxyl terminal residues (Phe(325), Ser(326) and Leu(327)) or mutation of the receptor (GnRHR[Phe(325)Ala]) caused a complete loss of measurable ligand binding and effector coupling, clearly suggesting an unexplained role for Phe(325).


Silica-Based Infiltrations for Enhanced Zirconia-Resin Interface Toughness.
Novel silica-based infiltrations on the surface of zirconia have the potential to improve their bondability, allowing for the etching/silane adhesive bonding technique. Nonetheless, adhesively bonded joints are subject to mixed tensile and shear stresses when the restoration is in occlusal service. Thus, we aimed to investigate the effect of 2 novel silica-based infiltrations on the interfacial toughness of adhesively bonded zirconia using the Brazil nut method, which allows for controlled types of stresses to be applied at the interfaces. In total, 150 3Y-TZP (In-Ceram YZ; Vita) Brazil nuts were machined and randomly assigned to 3 groups: C, control (air abraded); SG, sol-gel silica infiltration; and GI, glass infiltration. SG specimens were immersed twice in silicic acid for 20 min and dried (100°C, 1 h). GI specimens were presintered (1,400°C, 1 h) before a glass powder slurry was applied to the intaglio surface. All specimens were then sintered (1,530°C, 2 h). Following adhesive bonding (Panavia F 2.0, Kuraray) and water storage (37°C) for 10 d, the Brazil nuts were subdivided into groups baseline and aged (40,000 thermal cycles between 5°C and 55°C, with a dwell time of 30 s). The Brazil nuts were subjected to axial-loading tests using various inclinations (precrack angle with load direction): Θ = 0°, 5°, 10°, 15°, or 25°, which define the stress type at the interface, from pure tension (0°) to increasing levels of shear. Under pure tension (0°), GI yielded superior interfacial fracture energy, SG and C were similar, and aging had no effect. Under predominantly shear stresses (25°), aging significantly decreased interfacial fracture energy of C and SG, while GI remained stable and was superior. The glass infiltration of the zirconia intaglio surface increases its adhesive bonding interfacial toughness. The sol-gel silica infiltration method requires improvement to obtain a homogeneous surface infiltration and an enhanced bond strength.


Predicting Nodule Malignancy using a CNN Ensemble Approach.
Lung cancer is the leading cause of cancer-related deaths globally, which makes early detection and diagnosis a high priority. Computed tomography (CT) is the method of choice for early detection and diagnosis of lung cancer. Radiomics features extracted from CT-detected lung nodules provide a good platform for early detection, diagnosis, and prognosis. In particular when using low dose CT for lung cancer screening, effective use of radiomics can yield a precise non-invasive approach to nodule tracking. Lately, with the advancement of deep learning, convolutional neural networks (CNN) are also being used to analyze lung nodules. In this study, our own trained CNNs, a pre-trained CNN and radiomics features were used for predictive analysis. Using subsets of participants from the National Lung Screening Trial, we investigated if the prediction of nodule malignancy could be further enhanced by an ensemble of classifiers using different feature sets and learning approaches. We extracted probability predictions from our different models on an unseen test set and combined them to generate better predictions. Ensembles were able to yield increased accuracy and area under the receiver operating characteristic curve (AUC). The best-known AUC of 0.96 and accuracy of 89.45% were obtained, which are significant improvements over the previous best AUC of 0.87 and accuracy of 76.79%.


Effect of fortifiers and additional protein on the osmolarity of human milk: is it still safe for the premature infant?
The present guidelines of the American Academy of Pediatrics recommend that osmolarity not exceed 450 mOsm/kg (or approximately an osmolarity of 400 mOsm/L) for breast milk or infant formulae, to minimize the risk factors for necrotizing enterocolitis. A commercial protein supplement has been developed to meet special protein requirements (4.0-4.5 g · kg(-1) · day(-1)) of infants with a birth weight <1000 g. Because its effect on osmolarity has not been systematically studied, we characterized the effects of fortification on the osmolarity of human milk (HM). Osmolarity of fresh and processed HM was measured at baseline, after fortification with a commercial HM fortifier and after further supplementation with additional protein increasing in 0.5-g steps up to 4.0 g. Measurements were performed immediately after adding fortifier and/or protein and after 24 hours. In addition, changes in osmolarity were determined after adding therapeutic additives such as iron, multivitamin supplement, and calcium-phosphorus capsules. Native HM samples (n = 84) had 297 mOsm/L, (median; 95% confidence interval 295-299 mOsm/L). Adding HM fortifier increased osmolarity up to 436 mOsm/L (95% confidence interval 431-441 mOsm/L). Additional protein supplementation increased osmolarity by 23.5 mOsm/L per 0.5-g step, up to a maximum of 605 mOsm/L. Pasteurization decreased osmolarity by 20-30 mOsm/L (P < 0.001), and storage for 24 hours slightly increased osmolarity (by 11.5 mOsm/L P = 0.0002). Therapeutic additives increased osmolarity up to 868 mOsm/L. Adding HM fortifier and additional protein to HM increased osmolarity to >400 mOsm/L and therefore above the recommended threshold. Because of the excessive increase in osmolarity combinations of HM + fortifier and additional protein should not be applied together with multivitamins or other additives.


Risk-accepting personality and personal protective equipment use within the Agricultural Health Study.
Pesticide exposures can be reduced by use of personal protective equipment as well as proper mixing and application practices. The authors examined the effects of risk-accepting personality on personal protective equipment (PPE) use and mixing and application practices among private pesticide applicators and their spouses within the Agricultural Health Study (AHS) in Iowa and North Carolina and commercial applicators in Iowa. The AHS follow-up questionnaire included four questions designed to assess attitudes toward risk. Analysis was limited to those who were currently working on a farm or registered as a commercial applicator and indicated current pesticide use (n=25,166). Respondents who answered three or more questions in the affirmative (private applicators: n=4160 [21%]; commercial applicators: n=199 [14%]; spouses: n=829 [23%]) were classified as having a risk-accepting personality. Logistic regression was used to evaluate specific work practices associated with risk-accepting attitudes. Among private applicators, the likelihood of using any PPE when mixing or loading pesticides was lower among risk-acceptors compared to risk-averse individuals (odds ratio [OR] = 0.72, 95% confidence interval [CI]: 0.65-0.79). A similar relationship was observed among commercial applicators (OR = 0.77, 95% CI: 0.34-1.77) but not among spouses (OR = 1.09, 95% CI: 0.90-1.33). Among private applicators, risk-acceptors were more likely than the risk-averse to apply pesticides within 50 feet of the home (OR = 1.21, 95% CI: 1.01-1.44), compared to further than ¼ mile. These findings suggest that the decisions to use personal protective equipment and properly handle/apply pesticides may be driven by risk-accepting personality traits.


Violence by male partners against women during the childbearing year: a contextual analysis.
Many contextual analyses that bridge the micro-level-macro-level gap in identifying risk factors for adverse outcomes have not used methods appropriate for multilevel data. The purpose of this paper is to illustrate the application of appropriate multi-level analytic methods and discuss their implications for public health. A previously published individual-level model of physical violence perpetrated by male partners during the childbearing year was reanalyzed to include variables describing the neighborhoods where the women resided. Logistic regression with estimation methods of the generalized estimating equation was used for the contextual analysis. To assess the advantages of the generalized estimating equation over conventional logistic regression, both were used for the two-level model. The regression coefficients from the contextual model differed from the betas obtained in the individual-level model. Not only were neighborhood-level variables related to the risk of partner-perpetrated violence, but the presence of these macro-level variables in the models modified the relationships of the individual-level variables to the risk of violence. Two-level models that include individual- and community-level factors may be beneficial for purposes of explanation in public health research.


C-reactive protein and complement as acute phase reactants in common carp Cyprinus carpio during CyHV-3 infection.
Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of a highly virulent and lethal disease of common carp Cyprinus carpio and its ornamental koi varieties. However, specific knowledge about immune mechanisms behind the infection process is very limited. We aimed to evaluate the effect of the CyHV-3 infection on the profile of 2 major components of the common carp immune acute phase response: the C-reactive protein (CRP) and the complement system. Common carp were infected with CyHV-3 by bath immersion. Fish were sampled before the infection and at 6, 12, 24, 72, 120 and 336 h post-infection for serum and head kidney, liver, gill and spleen tissues. CRP levels and complement activity were determined from the serum, whereas CRP- and complement-related genes (crp1, crp2, c1rs, bf/c2, c3, masp2) expression profiles were analysed in the tissues by quantitative PCR. Both CRP levels and complement activity increased significantly up to 10- and 3-fold, respectively, in the serum of infected fish during the challenge. Analysis revealed distinct organ- and time-dependent expression profile patterns for all selected genes. These results suggest that CRP and complement behave as acute phase reactants to CyHV-3 infection in common carp with an organ- and time-dependent response.


Optimization of ribosome profiling using low-input brain tissue from fragile X syndrome model mice.
Dysregulated protein synthesis is a major underlying cause of many neurodevelopmental diseases including fragile X syndrome. In order to capture subtle but biologically significant differences in translation in these disorders, a robust technique is required. One powerful tool to study translational control is ribosome profiling, which is based on deep sequencing of mRNA fragments protected from ribonuclease (RNase) digestion by ribosomes. However, this approach has been mainly applied to rapidly dividing cells where translation is active and large amounts of starting material are readily available. The application of ribosome profiling to low-input brain tissue where translation is modest and gene expression changes between genotypes are expected to be small has not been carefully evaluated. Using hippocampal tissue from wide type and fragile X mental retardation 1 (Fmr1) knockout mice, we show that variable RNase digestion can lead to significant sample batch effects. We also establish GC content and ribosome footprint length as quality control metrics for RNase digestion. We performed RNase titration experiments for low-input samples to identify optimal conditions for this critical step that is often improperly conducted. Our data reveal that optimal RNase digestion is essential to ensure high quality and reproducibility of ribosome profiling for low-input brain tissue.


[Renal diseases related to MYH9 disorders].
Mutations in MYH9 gene encoding the nonmuscle myosin heavy chain IIA (NMMHC-IIA) are related to a number of rare autosomal-dominant disorders which has been known as May-Hegglin disease, Sebastian syndrome, Fechtner syndrome and Epstein syndrome. Their common clinical features are congenital macrothrombocytopaenia and polymorphonuclear inclusion bodies, in addition to a variable risk of developing proteinuria, chronic kidney disease progressing toward end stage, sensorineural deafness and presenile cataracts. The term MYH9 related disease (MYH9-RD) describes the variable expression of a single illness encompassing all previously mentioned hereditary disorders. Renal involvement in MYH9- RD has been observed in 30% of patients. Mutant MYH9 protein, expressed in podocytes, mesangial and tubular cells, plays a main role in foot process effacement and in development of nephropathy. Interestingly, the MYH9 gene is currently under investigation also for his possible contribution to many other non-hereditary glomerulopathies such as focal global glomerulosclerosis (hypertensive nephrosclerosis), idiopathic focal segmental glomerulosclerosis, C1q nephropathy and HIV-associated nephropathy. In this review we are aimed to describe renal diseases related to MYH9 disorders, from the hereditary disease to the acquired disorders, in which MYH9-gene acts as a "renal failure susceptibility gene".


Virus particles produced by the herpes simplex virus type 1 alkaline nuclease null mutant ambUL12 contain abnormal genomes.
Open reading frame UL12 of herpes simplex virus type 1 (HSV-1) encodes an alkaline nuclease that has previously been implicated in processing the complex, branched, viral DNA replication intermediates and allowing egress of DNA-containing capsids from the nucleus. This report describes experiments using the HSV-1 UL12 null mutant ambUL12, which aim to explain the approximately 200- to 1000-fold decrease in the yield of infectious virus, compared with wild-type (wt) HSV-1, from non-complementing cells. A detailed examination revealed that both DNA replication and encapsidation were affected in ambUL12-infected cells, resulting in an approximately 15- to 20-fold reduction in the amount of packaged DNA. In contrast to previous reports, the absence of UL12 function did not greatly impair capsid release into the cytoplasm, and virus particles were readily detected in the supernatant medium from ambUL12-infected cells. The released virus, however, exhibited much higher particle/p.f.u. ratios than wt HSV-1, and this made a further important contribution to the overall reduction in yield. Gel analyses of packaged ambUL12 and wt DNAs revealed the presence of structural abnormalities. The DNA obtained from extracellular ambUL12 virions was non-infectious in transfection assays, and both ambUL12 DNA and virus particles exerted a dominant inhibitory effect on the growth of wt virus. These results suggest that ambUL12 virions produced in non-complementing cells have a greatly reduced ability to initiate new cycles of infection, and that this defect results from the encapsidation of abnormal genomes.


Reaction mechanism of zinc-dependent cytosine deaminase from Escherichia coli: a quantum-chemical study.
The reaction mechanism of cytosine deaminase from Escherichia coli is studied using density functional theory. This zinc-dependent enzyme catalyzes the deamination of cytosine to form uracil and ammonia. The calculations give a detailed description of the catalytic mechanism and establish the role of important active-site residues. It is shown that Glu217 is essential for the initial deprotonation of the metal-bound water nucleophile and the subsequent protonation of the substrate. It is also demonstrated that His246 is unlikely to function as a proton shuttle in the nucleophile activation step, as previously proposed. The steps that follow are nucleophilic attack by the metal-bound hydroxide, protonation of the leaving group assisted by Asp313, and C-N bond cleavage. The calculated overall barrier is in good agreement with the experimental findings. Finally, the calculations reproduce the experimentally determined inverse solvent deuterium isotope effect, which further corroborates the suggested reaction mechanism.


The prevalence of femoroacetabular impingement anatomy in Division 1 aquatic athletes who tread water.
Femoroacetabular impingement (FAI) is a disorder that causes hip pain and disability in young patients, particularly athletes. Increased stress on the hip during development has been associated with increased risk of cam morphology. The specific forces involved are unclear, but may be due to continued rotational motion, like the eggbeater kick. The goal of this prospective cohort study was to use magnetic resonance imaging (MRI) to identify the prevalence of FAI anatomy in athletes who tread water and compare it to the literature on other sports. With university IRB approval, 20 Division 1 water polo players and synchronized swimmers (15 female, 5 male), ages 18-23 years (mean age 20.7 ± 1.4), completed the 33-item International Hip Outcome Tool and underwent non-contrast MRI scans of both hips using a 3 Tesla scanner. Recruitment was based on sport, with both symptomatic and asymptomatic individuals included. Cam and pincer morphology were identified. The Wilcoxon Signed-Rank/Rank Sum tests were used to assess outcomes. Seventy per cent (14/20) of subjects reported pain in their hips yet only 15% (3/20) sought clinical evaluation. Cam morphology was present in 67.5% (27/40) of hips, while 22.5% (9/40) demonstrated pincer morphology. The prevalence of cam morphology in water polo players and synchronized swimmers is greater than that reported for the general population and at a similar level as some other sports. From a clinical perspective, acknowledgment of the high prevalence of cam morphology in water polo players and synchronized swimmers should be considered when these athletes present with hip pain.


Inter-Rater Reliability of Preprocessing EEG Data: Impact of Subjective Artifact Removal on Associative Memory Task ERP Results.
The processing of EEG data routinely involves subjective removal of artifacts during a preprocessing stage. Preprocessing inter-rater reliability (IRR) and how differences in preprocessing may affect outcomes of primary event-related potential (ERP) analyses has not been previously assessed. Three raters independently preprocessed EEG data of 16 cognitively healthy adult participants (ages 18-39 years) who performed a memory task. Using intraclass correlations (ICCs), IRR was assessed for Early-frontal, Late-frontal, and Parietal Old/new memory effects contrasts across eight regions of interest (ROIs). IRR was good to excellent for all ROIs; 22 of 26 ICCs were above 0.80. Raters were highly consistent in preprocessing across ROIs, although the frontal pole ROI (ICC range 0.60-0.90) showed less consistency. Old/new parietal effects had highest ICCs with the lowest variability. Rater preprocessing differences did not alter primary ERP results. IRR for EEG preprocessing was good to excellent, and subjective rater-removal of EEG artifacts did not alter primary memory-task ERP results. Findings provide preliminary support for robustness of cognitive/memory task-related ERP results against significant inter-rater preprocessing variability and suggest reliability of EEG to assess cognitive-neurophysiological processes multiple preprocessors are involved.


Mechanisms of flunarizine-induced vasodilation in the rabbit mesenteric artery.
The vasodilating effects of flunarizine on smooth muscle strips of rabbit mesenteric artery have been investigated and compared with those of nifedipine. Flunarizine (30-300 nM) dose-dependently inhibited Ca2+-induced contractions in Ca2+-free solution containing 100 mM K+. Double reciprocal analysis showed that this inhibition was either competitive at low concentrations (30-100 nM; nifedipine-like) or noncompetitive at high concentrations (0.3-1 microM). The latter seemed to be partly related to an inhibition of contractile proteins as estimated from Ca2+-induced contractions in saponin-treated chemically skinned muscle strips. In contrast to the actions of nifedipine, flunarizine inhibited norepinephrine (NE)-induced contractions more than those induced by high K+, and at 0.3 microM, this agent totally blocked NE-induced contraction. Flunarizine also inhibited NE-induced contraction in Ca2+-free solution containing 2 mM EGTA. In Ca2+-free solution, NE rapidly hydrolyzed phosphatidylinositol 4,5-bisphosphate (PI-P2) and produced phosphatidic acid (PA). Flunarizine (30 and 300 nM), but not nifedipine (100 nM), inhibited NE-induced hydrolysis of PI-P2 and production of PA. However, flunarizine (100 nM) did not modify the contraction induced by 10 microM inositol 1,4,5-trisphosphate in chemically skinned muscle strips. It is concluded that flunarizine inhibits both voltage-dependent (nifedipine-like) and receptor-operated Ca2+ influx induced by NE and also inhibits NE-induced Ca2+ release from intracellular stores due to inhibition of the hydrolysis of PI-P2.


Sporting programs for inactive population groups: factors influencing implementation in the organized sports setting.
The organized sports sector has received increased attention as a setting to promote health-enhancing physical activity (HEPA) to the general population. For significant public health impact, it is important that successful HEPA programs are widely adopted, implemented and continued as ongoing practice. The importance of evaluating the context in which programs are implemented has been identified as critical. However, little research has focused on understanding the organized sports implementation context, including factors facilitating and impeding implementation. In this study, the main factors influencing implementation of HEPA programs in the organized sports setting were studied. Fourteen sporting programs in the Netherlands aimed at increasing participation in sports by inactive population groups and funded within the National Action Plan for Sport and Exercise (NAPSE) were investigated. The programs were developed by ten Dutch National Sports Federations (NSFs) and implemented by different sports clubs in the Netherlands over a 3-year implementation period (June 2008-June 2011). The qualitative research component involved yearly face-to-face interviews (i.e. fourteen interviews each year, n = 12 program coordinators) and a group meeting with the program coordinators of the NSFs (n = 8). Cross-case comparisons and thematic analyses were performed to identify and categorize important facilitating and impeding factors respectively. The quantitative research component, used to identify the most important facilitating and impeding factors across all sporting programs, consisted of ranking of factors according to importance by the program coordinators (n = 12). Different factors act during six identified (implementation) phases. When comparing factors across phases, several key learnings were evident. Successful implementation relied, for example, on program design and enthusiastic individuals within sporting organizations. On the other hand, inactive people were hard to reach and participation of sports clubs was not self-evident. The findings were discussed in a broader context. This study adds to the knowledge base concerning the implementation of sporting programs, aimed at inactive people, in the organized sports setting. The main factors facilitating and impeding implementation were identified. The results of this study can be used by sports practitioners and policy makers when developing and implementing HEPA programs in this setting.


Interactions between hypoxic and almitrine-induced vasoconstriction in the rat lung.
1. To test whether almitrine might improve the arterial partial pressure of O2 in patients with chronic obstructive airways disease by improvement of ventilation-perfusion matching, we looked at the interaction between hypoxic and almitrine-induced vasoconstriction in isolated rat lungs perfused with blood at constant flow. Increases in pressure represented increases in resistance. 2. Almitrine, given in increasing doses between challenges with 2% O2, enhanced hypoxic vasoconstriction at low doses but attenuated it at high doses. 3. Stimulus-response curves to hypoxia of increasing severity gave a sigmoid curve. 4. Almitrine solvent caused small changes in pulmonary artery pressure and shifted the stimulus-response curve slightly in a parallel fashion. 5. Small doses of almitrine enhanced the action of mild to moderate hypoxia, medium doses attenuated moderately severe hypoxia, whereas high doses depressed vasoconstriction due to all degrees of hypoxia. 6. These effects of almitrine on hypoxic vasoconstriction were compared with the effect of solvent by analysis of variance; the results substantiated significant enhancement of hypoxia by small doses and attenuation by large doses. 7. In patients, if similar effects apply, small doses of almitrine would assist ventilation-perfusion matching, but large doses might worsen it. 8. Almitrine-induced vasoconstriction was attenuated by a fall in perfusate temperature in a similar manner to hypoxic vasoconstriction. It was also attenuated by three drugs, chlorpheniramine, propanolol and diethylcarbamazine, all of which also decrease hypoxic vasoconstriction. The similarity between hypoxic and almitrine-induced pulmonary vasoconstriction is further confirmed.


Keeping the doors to the community open.
Polls show that Americans of all races now believe that social class and national origin are more potent bases for discrimination than race. However, this assessment is rendered more complicated by (a) the high rate of exclusion of people on the basis of behavioral characteristics and (b) the strikingly disparate impact of such decisions on people of color. America's high rates of (a) incarceration in the criminal and juvenile justice system, (b) visa denials, detention, and removal in the immigration system, and (c) suspensions and expulsions in the schools all have the effect of excluding many Latinos and African Americans from ordinary life in the community. Although norms of inclusion are adaptive for communities, such an approach is intrinsically difficult to achieve. Attention should be given to (a) facilitation of the exercise of moral leadership and (b) design of settings that ''demand'' notice of, and care for, all participants.


Dissemination of Klebsiella pneumoniae ST147 NDM-1 in Poland, 2015-19.
To assess the spread of New Delhi metallo-β-lactamase-1 (NDM-1)-producing Klebsiella pneumoniae ST147 organisms in Poland since an introduction from Tunisia in March 2015, including their phylogenetic position in the global population of the high-risk clone. Out of 8925 unique NDM-positive K. pneumoniae isolates identified in Poland from April 2015 till December 2019, 126 isolates, including the Tunisian imports, were related by PFGE and blaNDM gene-carrying Tn125 transposon derivatives. Forty-seven representative isolates were sequenced by Illumina MiSeq. The phylogeny, resistome, virulome and plasmid replicons were analysed and compared with the international ST147 strains. Plasmids of six isolates were studied by the MinION sequencing. A high homogeneity of the 47 isolates was observed, with minor variations in their resistomes and plasmid replicon profiles. However, the detailed SNP comparison discerned a strict outbreak cluster of 40 isolates. All of the organisms were grouped within the ST147 phylogenetic international lineage, and four NDM-1 producers from Tunisia, Egypt and France were the closest relatives of the Polish isolates. Yersiniabactin genes (YbST280 type) were located within the ICEKpn12-like element in most of the outbreak isolates, characterized by O2v1 and KL64 antigen loci. The blaNDM-1 genes were located in double-replicon IncFIIK2+IncFIBK plasmids. The continuous spread of K. pneumoniae ST147 NDM-1 in Poland since 2015, largely in the Warsaw area, is demonstrated by this genomic analysis. The isolates showed a high degree of homogeneity, and close relatedness to organisms spreading in the Mediterranean region.


Traumatic brain injury and post-acute decline: what role does environmental enrichment play? A scoping review.
While a growing number of studies provide evidence of neural and cognitive decline in traumatic brain injury (TBI) survivors during the post-acute stages of injury, there is limited research as of yet on environmental factors that may influence this decline. The purposes of this paper, therefore, are to (1) examine evidence that environmental enrichment (EE) can influence long-term outcome following TBI, and (2) examine the nature of post-acute environments, whether they vary in degree of EE, and what impact these variations have on outcomes. We conducted a scoping review to identify studies on EE in animals and humans, and post-discharge experiences that relate to barriers to recovery. One hundred and twenty-three articles that met inclusion criteria demonstrated the benefits of EE on brain and behavior in healthy and brain-injured animals and humans. Nineteen papers on post-discharge experiences revealed that variables such as insurance coverage, financial, and social support, home therapy, and transition from hospital to home, can have an impact on clinical outcomes. There is evidence to suggest that lack of EE, whether from lack of resources or limited ability to engage in such environments, may play a role in post-acute cognitive and neural decline. Maximizing EE in the post-acute stages of TBI may improve long-term outcomes for the individual, their family and society.


NTP Toxicity Studies of 60-Hz Magnetic Fields Administered by Whole Body Exposure to F344/N Rats, Sprague-Dawley Rats, and B6C3F1 Mice.
Electric and magnetic fields are associated with the production, transmission, and use of electricity; thus the potential for human exposure is high. These electric and magnetic fields are predominantly of low frequency (60 Hz) and generally of low intensity. The prevailing view among physicists is that exposure to these low-frequency, low-intensity fields does not pose a health hazard. However, this view has been challenged by reports linking magnetic field exposure to the development of leukemia and other cancers. Because multiple epidemiologic studies suggested a potential for increased cancer rates with increasing exposure, and because of public concern, the effects of 60-Hz magnetic field exposure were examined in F344/N rats and B6C3F1 mice in 8-week full-body-exposure studies. Animals were evaluated for hematology and clinical chemistry (rats only) parameters, pineal gland hormone concentrations, and histopathology. Additional studies were performed in Sprague-Dawley rats to examine teratologic and reproductive effects of magnetic field exposure. In the 8-week toxicity studies, groups of male and female F344/N rats and B6C3F1 mice were exposed for 18.5 hours per day to 60-Hz magnetic fields at intensities of 0 (control), 0.02, 2, and 10 gauss (G). Additional groups of rats and mice were exposed to intermittent 10 G fields (1 hour on/1 hour off) for 18.5 hours per day. No evidence of toxicity associated with exposure to magnetic fields was observed in rats or mice. Clinical observations provided no evidence of adverse effects associated with magnetic field exposure. Compared to control rats and mice, there were no biologically significant differences in hematology or clinical chemistry parameters of rats exposed to magnetic fields. No gross lesions or histopathologic findings in rats or mice were attributed to exposure to 60-Hz magnetic fields. In addition, magnetic field exposure was not associated with a significant reduction in serum melatonin or pineal gland melatonin concentration, or pineal gland activity of N-acetyltransferase in either species. One female rat in the 2 G exposure group died during the 8-week toxicity study from causes unrelated to magnetic field exposure; all other male and female rats and mice in the study survived until the end of the study. Final mean body weights and mean organ weights of a few groups of exposed animals differed from those of the control groups; however, no clear pattern of magnetic field effects was observed, and these differences are not considered to be biologically significant. For the teratology study, groups of 55 pregnant female Sprague-Dawley rats were exposed to the same magnetic fields as in the toxicity study on gestation days 6 through 19. Fifteen pregnant females exposed to 85 mg ethylenethiourea/kg body weight served as positive controls. Except for the positive controls, there were no changes in maternal or fetal weights, nor were fetal abnormalities found. The number of pregnant females was significantly lower in groups exposed to magnetic fields than in the control group; however, all breeding in the teratology study was completed prior to the first day of magnetic field or sham exposure. On this basis, this finding is unrelated to magnetic field exposure. In addition, there were no differences between control and exposed groups in the number of pregnant females in the continuous breeding study, in which breeding took place within the magnetic fields. Groups of 40 breeding pairs of Sprague-Dawley rats were exposed to the same magnetic fields as in the toxicity study during the breeding and lactation of five litters in a continuous breeding study. The fifth litter was exposed during gestation and lactation; one male and one female from each litter were raised to sexual maturity receiving the same exposures as the parents; rats were mated to nonsibling rats and allowed to deliver the third-generation offspring. The results of the continuous breeding study demonstrated no effects of magnetic field exposure on reproductive performance in either male or female rats.


Muscle Rad expression and human metabolism: potential role of the novel Ras-related GTPase in energy expenditure and body composition.
Ras associated with diabetes (Rad), a new ras-related GTPase, was recently identified by subtractive cloning as an mRNA in skeletal muscle that is overexpressed in NIDDM. To better understand its metabolic significance, we measured skeletal muscle Rad expression in well-characterized insulin sensitive (IS) and insulin resistant (IR) subjects with normal glucose tolerance and in untreated NIDDM patients. We found no differences in expression of Rad mRNA levels among IS, IR, and NIDDM groups using a ribonuclease protection assay (0.22 +/- 0.06, 0.13 +/- 0.01, and 0.16 +/- 0.02 relative units, respectively; NS) and no differences in Rad protein expression using a specific anti-peptide Rad antibody (1.05 +/- 0.18, 1.14 +/- 0.08, and 1.08 +/- 0.21 units/mg protein, respectively; NS). However, Rad protein levels were positively correlated with BMI (r = 0.43, P = 0.03) and percentage body fat (r = 0.55, P < 0.005), two independent measures of obesity, and negatively correlated with resting metabolic rate (r = 0.49, P = 0.01). In multiple regression analyses, percentage body fat and resting metabolic rate independently accounted for 30 and 10% of individual variability in muscle Rad protein expression. In conclusion, Rad expression in skeletal muscle is not altered as a function of insulin resistance or NIDDM in humans. However, these data, for the first time, implicate a role for Rad in regulating body composition and energy expenditure and provide a framework for studies designed to elucidate Rad's cellular functions.


Countering countermeasures: detecting identity lies by detecting conscious breakthrough.
One major drawback of deception detection is its vulnerability to countermeasures, whereby participants wilfully modulate their physiological or neurophysiological response to critical guilt-determining stimuli. One reason for this vulnerability is that stimuli are usually presented slowly. This allows enough time to consciously apply countermeasures, once the role of stimuli is determined. However, by increasing presentation speed, stimuli can be placed on the fringe of awareness, rendering it hard to perceive those that have not been previously identified, hindering the possibility to employ countermeasures. We tested an identity deception detector by presenting first names in Rapid Serial Visual Presentation and instructing participants to lie about their own identity. We also instructed participants to apply a series of countermeasures. The method proved resilient, remaining effective at detecting deception under all countermeasures.


Metabolic activation of N-hydroxy arylamines and N-hydroxy heterocyclic amines by human sulfotransferase(s).
Several N-hydroxy metabolites of carcinogenic arylamines and heterocyclic amines were examined as substrates for bioactivation by human liver sulfotransferases (STs). Among the N-hydroxy derivatives studied, N-hydroxy-2-acetylaminofluorene, N-hydroxy-2-aminofluorene, N-hydroxy-4,4'-methylene-bis(2-chloroaniline), N-hydroxy-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, and N-hydroxy-2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole were each metabolically activated by 3'-phosphoadenosine-5'-phosphosulfate-dependent human liver STs. No ST-mediated DNA binding of N-hydroxy-2-amino-3-methylimidazo[4,5-f]quinoline or N-hydroxy-2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline was detected under our assay conditions. In the 12 human hepatic cytosols studied, the extent of 3'-phosphoadenosine-5'-phosphosulfate-dependent DNA binding of the N-hydroxy derivatives were all significantly correlated with levels of thermostable phenol ST (TS-PST) activity but not with thermolabile phenol ST or dehydroepiandrosterone ST activities. The propensity of these N-hydroxy arylamines and N-hydroxy heterocyclic amines to serve as selective substrates for human TS-PST was further confirmed by inhibition with 2,6-dichloro-4-nitrophenol and by thermostability studies. N-hydroxy-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine and N-hydroxy-4,4'-methylene-bis(2-chloroaniline) were also used as substrates to study ST-dependent metabolic activation in other human tissue preparations. 3'-phosphoadenosine-5'-phosphosulfate-dependent DNA binding activity was detected in human liver and colon cytosols but not in pancreas, larynx, or urinary bladder epithelial cytosols. Since the TS-PST appears to be expressed polymorphically in human populations, the finding that human TS-PST is capable of metabolically activating N-hydroxy metabolites of several carcinogenic arylamines and heterocyclic amines suggests that TS-PST may have an important role in determining interindividual susceptibility to these environmental and dietary carcinogens.


Tumorigenicity in athymic mice of the human colon carcinoma cell line SW1116 expressing the tumor-associated antigenic determinant CA 19-9.
The human colorectal carcinoma cell line SW1116 under optimal growth conditions synthesized and shed antigens bearing the monoclonal antibody-defined carbohydrate determinant CA 19-9. Antigen expressing CA 19-9 in cell culture supernatant was quantitated by an immunoradiometric assay for CA 19-9. Injection of SW1116 cells s.c. into athymic BALB/c mice resulted in the growth of moderately differentiated tumors possessing a distinct morphological resemblance to a typical adenocarcinoma of the colon. Intervals to tumor appearance were dependent on inoculum dose, but 95% of mice at both 5 X 10(6) and 10(7) cells/mouse developed tumors within 14 to 21 days. CA 19-9 antigen was detected in the sera of all nude mice with SW1116 tumors, and antigen concentration correlated (r = 0.77) with tumor volume throughout the 9-week study. The half-life of this antigen in serum following tumor excision from nude mice was 6.5 +/- 1.5 (S.D.) hr. Carcinoembryonic antigen was also detected in serum from mice bearing SW1116 tumors by an immunoradiometric assay for carcinoembryonic antigen, but its concentration correlated (r = 0.86) with tumor volume for only the first 4 weeks of tumor growth. Significant levels of endogenous immunoglobulin G1 and immunoglobulin G3 antibodies to CA 19-9 antigen were found in the serum of nude mice with SW1116 tumors by radioimmunodiffusion, but no apparent relationship between antibody titer and tumor growth or CA 19-9 antigen level in serum was evident. This tumor model may be useful in devising radioimmunodetection and immunotherapeutic strategies for primary and metastatic human colon carcinomas.


Mediating mechanisms for the impact of bupropion in smoking cessation treatment.
Several studies have documented the effectiveness of bupropion for smoking cessation, yet little is known about the mechanisms by which it facilitates abstinence. In this placebo-controlled randomized trial. We examined whether bupropion's effects on cessation were mediated by changes in withdrawal and/or negative or positive affect (PA). Two hundred and fifty-one smokers received 10-week treatment with bupropion or placebo, plus behavioral counseling. Changes in affect and withdrawal symptoms from pre-quit to 1 week post quit were examined as mediating variables in structural equation models. Cotinine-verified 7-day point prevalence cessation rates at the end of treatment (8-weeks post quit date) were 48% for bupropion and 29% for placebo (P=0.001). There were significant treatment effects on withdrawal and negative affect (NA); however, only change in NA predicted cessation. In a path model, change in NA was a significant mediator of bupropion's effects on cessation. However, the proportion of variance accounted for by this mediator was small, suggesting that other unmeasured factors play an important role. Laboratory-based paradigms may be useful to identify other mediators of bupropion's effects, thereby pointing to mechanisms of effect that can be bolstered in future treatment studies.


Molecularly imprinted polymer cartridges coupled on-line with high performance liquid chromatography for simple and rapid analysis of human insulin in plasma and pharmaceutical formulations.
In this paper, a novel method is described for automated determination of human insulin in biological fluids using principle of sequential injection on a molecularly imprinted solid-phase extraction (MISPE) cartridge as a sample clean-up technique combined with high performance liquid chromatography (HPLC). The water-compatible molecularly imprinted polymers (MIPs) were prepared using methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross-linker, chloroform as a porogen and insulin as a template molecule. The imprinted polymers were then employed as the solid-phase extraction sorbent for on-line extraction of insulin from human plasma samples. To achieve the best condition, influential parameters on the extraction efficiency were thoroughly investigated. Rapid and simple analysis of the hormone was successfully accomplished through the good selectivity of the prepared sorbent coupled with HPLC. Limits of detection (LOD) and quantification (LOQ) of 0.2 ng mL(-1), 0.7 ng mL(-1), and 0.03 ng mL(-1), 0.1 ng mL(-1) were obtained in plasma and urine respectively. The obtained data exhibited the great recoveries for extraction of insulin from human plasma and pharmaceutical samples, higher than 87%.


Functional limitations and overweight among migrants in the Histoire de Vie study (Insee, 2003).
In France, epidemiological data on immigrant population are limited since characterization of immigrants is viewed as a sensitive issue. The Histoire de Vie survey has enabled the description of the association of immigration characteristics and two health indicators. Health-related functional limitations and overweight status (BMI> or =25 kg/m(2)) were studied among 18 to 64 years old using indirect age standardization. Immigration status was characterized in four different ways: via the migratory pathways of interviewees and their parents (French born to two French-born parents, second-generation, immigrated at the age of 15 or younger, immigrated at the age of 16 or older), citizenship (French by birth, French by naturalisation, non French), geographical origin (France, Europe, outside Europe) and language(s) spoken at home (French mother tongue, French for persons who do not have French as mother tongue, French and another language, another language). The analyses were conducted separately in men and in women and stratified by occupational status (upper and lower). For functional limitations, the analysis was repeated excluding participants whose limitations occurred before the age of 19, which was the average age of immigration, to assess a possible healthy migrant effect. Compared to French men born to two French parents, an increased rate of functional limitations was observed among men born in Europe and/or who had at least one parent born in Europe (SMR: 1.4; 95% CI: 1.06-1.81) and a reduced rate among participants born outside Europe or who had at least one parent born outside Europe (SMR: 0.63; 95% CI: 0.46-0.86); these differences remained when social status was taken into account. Among women, a tendency towards a lower rate of functional limitations was found in lower social status groups among those who immigrated as adults, among non French citizens and among non Europeans. Restricting the analysis to individuals free of limitations at the age of 19 did not suggest a healthier migrant effect. Among men, the risk of being overweight did not differ according to immigration characteristics. Women who immigrated in their adult life (SMR: 1.42; 95% CI: 1.18-1.74), who had foreign citizenship (SMR: 1.44; 95% CI: 1.18-1.74) and who still spoke their mother tongue at home (SMR: 1.53; 95%CI: 1.23-1.88) were more likely to be overweight than non migrant women. These results were observed in lower social groups but not in upper social groups. These results demonstrate a heterogeneous relationship between immigration status and health, according to age at immigration, gender and origin while the migrant second-generation appears quite close to the population born French to two French-born parents. Further studies are needed to provide data on a wider range of health indicators. In the future, health surveys should document detailed information to allow for a full characterisation of the migrant population.


Altering the reaction coordinate of the ATP sulfurylase-GTPase reaction.
ATP sulfurylase, isolated from Escherichia coli K-12, catalyzes and couples two reactions: the hydrolysis of GTP and the synthesis of APS (adenosine 5'-phosphosulfate). Its GTPase activity is regulated in response to ligand binding at the APS-forming active site. In particular, AMP mimics an intermediate-like form of the enzyme that increases the k(cat) for GTP hydrolysis 180-fold. Using equilibrium and pre-steady-state methods, we have determined the relative Gibbs energies for many of the ground and transition states in the GTPase catalytic cycle, in the presence and absence of AMP. GTP and AMP energetically interact throughout the substrate branch of the reaction coordinate; however, once bond breaking occurs, communication between nucleotides ceases. Stopped-flow experiments, using the fluorescent nucleotides 2'-deoxy-mant-GTP and -GDP, indicate that the binding of AMP fosters a conformation of the enzyme that hinders the addition of 2'-deoxy-mant-GTP into the active site without affecting its escaping tendency. These results explain the effects of AMP on the equilibrium binding of the 2'-deoxy-mant-GTP. The second-order rate constants for the binding of 2'-deoxy-mant-GTP or -GDP, approximately 1 x 10(-6) M(-1) s(-1), are 2-3 orders of magnitude less than expected for simple diffusion models, and the binding progress curves appear biphasic. These findings suggest the presence of an intermediate(s) in the binding reactions. The Gibbs energy changes that occur in the reaction coordinate upon binding of AMP clearly show that the catalytic effect of AMP is due primarily to its -3.1 kcal/mol stabilization of the rate-limiting transition state.


Phosphorylation of Skp2 regulated by CDK2 and Cdc14B protects it from degradation by APC(Cdh1) in G1 phase.
The p27(Kip1) ubiquitin ligase receptor Skp2 is often overexpressed in human tumours and displays oncogenic properties. The activity of SCF(Skp2) is regulated by the APC(Cdh1), which targets Skp2 for degradation. Here we show that Skp2 phosphorylation on Ser64/Ser72 positively regulates its function in vivo. Phosphorylation of Ser64, and to a lesser extent Ser72, stabilizes Skp2 by interfering with its association with Cdh1, without affecting intrinsic ligase activity. Cyclin-dependent kinase (CDK)2-mediated phosphorylation of Skp2 on Ser64 allows its expression in mid-G1 phase, even in the presence of active APC(Cdh1). Reciprocally, dephosphorylation of Skp2 by the mitotic phosphatase Cdc14B at the M --> G1 transition promotes its degradation by APC(Cdh1). Importantly, lowering the levels of Cdc14B accelerates cell cycle progression from mitosis to S phase in an Skp2-dependent manner, demonstrating epistatic relationship of Cdc14B and Skp2 in the regulation of G1 length. Thus, our results reveal that reversible phosphorylation plays a key role in the timing of Skp2 expression in the cell cycle.


Measurement of Systemic Mitochondrial Function in Advanced Primary Open-Angle Glaucoma and Leber Hereditary Optic Neuropathy.
Primary Open Angle Glaucoma (POAG) is a common neurodegenerative disease characterized by the selective and gradual loss of retinal ganglion cells (RGCs). Aging and increased intraocular pressure (IOP) are glaucoma risk factors; nevertheless patients deteriorate at all levels of IOP, implying other causative factors. Recent evidence presents mitochondrial oxidative phosphorylation (OXPHOS) complex-I impairments in POAG. Leber Hereditary Optic Neuropathy (LHON) patients suffer specific and rapid loss of RGCs, predominantly in young adult males, due to complex-I mutations in the mitochondrial genome. This study directly compares the degree of OXPHOS impairment in POAG and LHON patients, testing the hypothesis that the milder clinical disease in POAG is due to a milder complex-I impairment. To assess overall mitochondrial capacity, cells can be forced to produce ATP primarily from mitochondrial OXPHOS by switching the media carbon source to galactose. Under these conditions POAG lymphoblasts grew 1.47 times slower than controls, whilst LHON lymphoblasts demonstrated a greater degree of growth impairment (2.35 times slower). Complex-I enzyme specific activity was reduced by 18% in POAG lymphoblasts and by 29% in LHON lymphoblasts. We also assessed complex-I ATP synthesis, which was 19% decreased in POAG patients and 17% decreased in LHON patients. This study demonstrates both POAG and LHON lymphoblasts have impaired complex-I, and in the majority of aspects the functional defects in POAG were milder than LHON, which could reflect the milder disease development of POAG. This new evidence places POAG in the spectrum of mitochondrial optic neuropathies and raises the possibility for new therapeutic targets aimed at improving mitochondrial function.


Development and validation of a quantitative snack and beverage food frequency questionnaire for adolescents.
A short, reliable and valid tool to measure snack and beverage consumption in adolescents, taking into account the correct definitions, would benefit both epidemiological and intervention research. The present study aimed to develop a short quantitative beverage and snack food frequency questionnaire (FFQ) and to assess the reliability and validity of this FFQ against three 24-h recalls. Reliability was assessed by comparing estimates of the FFQ administered 14 days apart (FFQ1 and FFQ2) in a convenience sample of 179 adolescents [60.3% male; mean (SD) 14.7 (0.9) years]. Validity was assessed by comparing FFQ1 with three telephone-administered 24-h recalls in a convenience sample of 99 adolescents [52.5% male, mean (SD) 14.8 (0.9) years]. Reliability and validity were assessed using Bland-Altman plots, classification agreements and correlation coefficients for the amount and frequency of consumption of unhealthy snacks, healthy snacks, unhealthy beverages, healthy beverages, and for the healthy snack and beverage ratios. Small mean differences (FFQ1 versus FFQ2) were observed for reliability, ranking ability ranged from fair to substantial, and Spearman coefficients fell within normal ranges. For the validity, mean differences (FFQ1 versus recalls) were small for beverage intake but large for snack intake, except for the healthy snack ratio. Ranking ability ranged from slightly to moderate, and Spearman coefficients fell within normal ranges. Reliability and validity of the FFQ for all outcomes were found to be acceptable at a group level for epidemiological purposes, whereas for intervention purposes only the healthy snack and beverage ratios were found to be acceptable at a group level.