Pillararene-based fluorescent chemosensors: recent advances and perspectives.
In 2008, a new class of pillar-shaped supramolecular macrocyclic hosts was reported, known as "pillararenes". Their particular electron-rich cavity and the ease of their functionalization offer possibilities for the design and synthesis of novel fluorescent chemosensors. Subsequently, pillararene-based fluorescent sensors and probes have been rapidly developed. This feature article covers the most recent contributions from the pillararene-based fluorescent sensor field in terms of anion/cation sensing, small molecule recognition, biomolecule detection, fluorescent supramolecular aggregates, and biomedical imaging. Meanwhile, we hope that this feature article will inspire more effort to be devoted to this emerging field.


Quantitative Assessment via Multi-Domain Fusion of Muscle Synergy Associated with Upper-Limb Motor Function for Stroke Rehabilitation.
Quantitative assessment of upper limb motor function can assist therapists in providing appropriate rehabilitation strategies, which plays an essential role in post-stroke rehabilitation. Conventionally, the most frequently used assessments are based on clinical scales or kinematic metrics, which rely on subjective scores or may be masked at the kinematic level by compensatory strategies. Recently, muscle synergies which encodes the simplified neuromuscular control strategy deployed by the central nervous system have been gradually used to assess post-stroke impairment. In general, muscle synergies are decomposed into two components: synergy vectors and synergy activation. Synergy vectors represent the relative weighting of each muscle within each synergy, that is muscle coordination; synergy activation represents the recruitment of the muscle synergy over time, that is muscle activation strength. Both the characteristics of synergy vectors and synergy activation are crucial for adequately assessing patients' motor function. Therefore, we integrate the spatial domain and temporal domain features extracted from synergy vectors and synergy activation for constructing a multi-domain assessment system based on Random Forest classifier, which may provide great qualitative classification accuracy. Furthermore, a novel functional score is generated from the probabilities belonging to the pathological group. Finally, we conduct a study with ten healthy subjects and ten post-stroke patients to verify the effectiveness of the proposed method. The experimental results show that the classification accuracy was enhanced to 98.56% by fusing the characteristics derived from different domains, which was higher than that based on spatial domain (94.90%) and temporal domain (91.08%), respectively. Furthermore, the assessment score generated by multi-domain fusion framework exhibited a significant correlation with the clinical score. These promising results show the potential of applying the proposed method to clinical assessments for post-stroke patients.


The implications of methylphenidate use by healthy medical students and doctors in South Africa.
The use of medical stimulants to sustain attention, augment memory and enhance intellectual capacity is increasing in society. The use of Methylphenidate for cognitive enhancement is a subject that has received much attention in the literature and academic circles in recent times globally. Medical doctors and medical students appear to be equally involved in the off-label use of Methylphenidate. This presents a potential harm to society and the individual as the long-term side effect profile of this medication is unknown. The implication of the use of Methylphenidate by medical students and doctors has not been fully explored. This article considers the impact of this use on the traditional role of medicine, society, the patient and suggests a way forward. We discuss the salient philosophy surrounding the use of cognitive enhancement. We query whether there are cognitive benefits to the use of Methylphenidate in healthy students and doctors and whether these benefits would outweigh the risks in taking the medication. Could these benefits lead to tangible outcomes for society and could the off label-use of Methylphenidate potentially undermine the medical profession and the treatment of patients? If cognitive benefits are proven then doctors may be coerced explicitly or implicitly to use the drug which may undermine their autonomy. The increased appeal of cognitive enhancement challenges the traditional role of medicine in society, and calls into question the role of a virtuous life as a contributing factor for achievement. In countries with vast economic disparity such as South Africa an enhancement of personal utility that can be bought may lead to greater inequities. Under the status quo the distribution of methylphenidate is unjust. Regulatory governmental policy must seek to remedy this while minimising the potential for competitive advantage for the enhanced. Public debate on the use of cognitive enhancement is long overdue and must be stimulated. The use of Methylphenidate for cognitive enhancement is philosophically defendable if long-term research can prove that the risks are negligible and the outcomes tangible.


Novel transient scattering technique for femtosecond dephasing measurements.
A novel transient scattering technique is proposed for femtosecond dephasing time measurements. This technique provides several advantages compared with existing techniques for time-domain dephasing measurements, including resolution below the pulse width, insensitivity to rapid energy relaxation, and clear demarcation between homogeneous and inhomogeneous broadening. We report experimental results for the dye Nile blue in solution; the dephasing time T(2) is determined to be less than 20 fsec.


Biometric and mobile gait analysis for early diagnosis and therapy monitoring in Parkinson's disease.
Parkinson's disease (PD) is the most frequent neurodegenerative movement disorder. Early diagnosis and effective therapy monitoring is an important prerequisite to treat patients and reduce health care costs. Objective and non-invasive assessment strategies are an urgent need in order to achieve this goal. In this study we apply a mobile, lightweight and easy applicable sensor based gait analysis system to measure gait patterns in PD and to distinguish mild and severe impairment of gait. Examinations of 16 healthy controls, 14 PD patients in an early stage, and 13 PD patients in an intermediate stage were included. Subjects performed standardized gait tests while wearing sport shoes equipped with inertial sensors (gyroscopes and accelerometers). Signals were recorded wirelessly, features were extracted, and distinct subpopulations classified using different classification algorithms. The presented system is able to classify patients and controls (for early diagnosis) with a sensitivity of 88% and a specificity of 86%. In addition it is possible to distinguish mild from severe gait impairment (for therapy monitoring) with 100% sensitivity and 100% specificity. This system may be able to objectively classify PD gait patterns providing important and complementary information for patients, caregivers and therapists.


Theory of diffusion-influenced reactions in complex geometries.
Chemical transformations involving the diffusion of reactants and subsequent chemical fixation steps are generally termed "diffusion-influenced reactions" (DIR). Virtually all biochemical processes in living media can be counted among them, together with those occurring in an ever-growing number of emerging nano-technologies. The role of the environment's geometry (obstacles, compartmentalization) and distributed reactivity (competitive reactants, traps) is key in modulating the rate constants of DIRs, and is therefore a prime design parameter. Yet, it is a formidable challenge to build a comprehensive theory that is able to describe the environment's "reactive geometry". Here we show that such a theory can be built by unfolding this many-body problem through addition theorems for special functions. Our method is powerful and general and allows one to study a given DIR reaction occurring in arbitrary "reactive landscapes", made of multiple spherical boundaries of given size and reactivity. Importantly, ready-to-use analytical formulas can be derived easily in most cases.


Cortical Response to Fat Taste.
We sense fat by its texture and smell, but it is still unknown whether we also taste fat despite evidence of both candidate receptors and distinct fat taste sensations. One major reason fat is still not recognized as a basic taste quality is that we first need to demonstrate its underlying neural activity. To investigate such neural fat taste activation, we recorded evoked responses to commercial cow milk products with 0.1%, 4%, and 38 % fat via high-density electroencephalography (EEG) from 24 human participants. The experimental design ensured that the products would only be discriminable via their potential fat taste; all stimuli were carefully controlled for differences in viscosity, lubrication, odor, temperature, and confounding tastes (sweetness, acidity, and "off-taste") and were delivered directly onto the tongue using a set of computer-controlled syringe pumps. Advanced topographical pattern analysis revealed different neural activation to the milk products 85-134 ms after stimulus onset, which, as expected, best discriminated the two milk fat extremes (0.1% and 38% fat). Notably, this time period has previously been shown to also encode basic taste qualities, such as sweet or salty. By adding to the evidence of cortical fat taste processing in response to staple food, our finding not only substantiates that we taste fat but also highlights its potential relevance during our everyday lives with possible large-scale impacts on motivational eating behavior to explain overconsumption of energy-dense foods.


Angiomotin regulates visceral endoderm movements during mouse embryogenesis.
In pregastrula stage mouse embryos, visceral endoderm (VE) migrates from a distal to anterior position to initiate anterior identity in the adjacent epiblast. This anterior visceral endoderm (AVE) is then displaced away from the epiblast by the definitive endoderm to become associated with the extra-embryonic ectoderm and subsequently contributes to the yolk sac. Little is known about the molecules that regulate this proximal displacement. Here we describe a role for mouse angiomotin (amot) in VE movements. amot expression is initially detected in the AVE and subsequently in the VE associated with the extra-embryonic ectoderm. Most amot mutant mice die soon after gastrulation with distinct furrows of VE located at the junction of the embryonic and extra-embryonic regions. Mutant analysis suggests that VE accumulation in these furrows is caused by defects in cell migration into proximal extra-embryonic regions, although distal-to-anterior movements associated with the epiblast, definitive endoderm formation, and anterior specification of the epiblast appear to be normal. These results suggest that amot acts within subregions of the VE to regulate morphogenetic movements that are required for embryo viability.


N-3 fatty acid rich triglyceride emulsions are neuroprotective after cerebral hypoxic-ischemic injury in neonatal mice.
We questioned if acute administration of n-3 fatty acids (FA) carried in n-3 rich triglyceride (TG) emulsions provides neuroprotection in neonatal mice subjected to hypoxic-ischemic (H/I) brain injury. We examined specificity of FA, optimal doses, and therapeutic windows for neuroprotection after H/I. H/I insult was induced in C57BL/6J 10-day-old mice by right carotid artery ligation followed by exposure to 8% O(2) for 15 minutes at 37°C. Intraperitoneal injection with n-3-rich TG emulsions, n-6 rich TG emulsions or saline for control was administered at different time points before and/or after H/I. In separate experiments, dose responses were determined with TG containing only docosahexaenoic acid (Tri-DHA) or eicosapentaenoic acid (Tri-EPA) with a range of 0.1-0.375 g n-3 TG/kg, administered immediately after H/I insult. Infarct volume and cerebral blood flow (CBF) were measured. Treatment with n-3 TG emulsions both before- and after- H/I significantly reduced total infarct volume by a mean of 43% when administered 90 min prior to H/I and by 47% when administered immediately after H/I. In post-H/I experiments Tri-DHA, but not Tri-EPA exhibited neuroprotective effects with both low and high doses (p<0.05). Moreover, delayed post-H/I treatment with Tri-DHA significantly decreased total infarct volume by a mean of 51% when administered at 0 hr, by 46% at 1 hr, and by 51% at 2 hr after H/I insult. No protective effect occurred with Tri-DHA injection at 4 hr after H/I. There were no n-3 TG related differences in CBF. A significant reduction in brain tissue death was maintained after Tri-DHA injection at 8 wk after the initial brain injury. Thus, n-3 TG, specifically containing DHA, is protective against H/I induced brain infarction when administered up to 2 hr after H/I injury. Acute administration of TG-rich DHA may prove effective for treatment of stroke in humans.


Culture expansion in low-glucose conditions preserves chondrocyte differentiation and enhances their subsequent capacity to form cartilage tissue in three-dimensional culture.
Culture conditions that preserve a stable chondrocyte phenotype are desirable in cell-based cartilage repair to maximize efficacy and clinical outcome. This study investigates whether low-glucose conditions will preserve the chondrocyte phenotype during culture expansion. Articular chondrocytes were culture-expanded in media supplemented with either low (1 mM) or high (10 mM) glucose. The metabolic phenotype, reactive oxygen species generation, and mRNA expression of markers of differentiation or catabolism were assessed by reverse-transcription quantitative polymerase chain reaction after four population doublings (PDs) and subsequent tissue formation capacity determined using pellet cultures. Continuous monolayer culture was used to determine the population doubling limit. After expansion in monolayer for four PDs, chondrocytes expanded in low-glucose conditions exhibited higher expression of the differentiation markers SOX9 and COL2A1 and reduced expression of the catabolic metalloproteinase matrix metallopeptidase 13. When chondrocytes expanded in low glucose were cultured in micropellets, they consistently generated more cartilaginous extracellular matrix than those expanded in high glucose, as evaluated by wet weight, sulfated glycosaminoglycan content, and hydroxyproline assay for collagen content. The same pattern was observed whether high or low glucose was used during the pellet culture. During expansion, chondrocytes in high-glucose generated 50% more reactive oxygen species than low-glucose conditions, despite a lower dependence on oxidative phosphorylation for energy. Furthermore low-glucose cells exhibited >30% increased population doubling limit. These data suggests that low-glucose expansion conditions better preserve the expression of differentiation markers by chondrocytes and enhance their subsequent capacity to form cartilage in vitro. Therefore, low glucose levels should be considered for the expansion of chondrocytes intended for tissue engineering applications.


Almonds and Cardiovascular Health: A Review.
Several preventive strategies to reduce dyslipidemia have been suggested, of which dietary modification features as an important one. Dyslipidemia is a major risk factor for coronary heart disease and strategies to manage dyslipidemia have been shown to reduce the incidence of cardiovascular disease (CVD). Although there are proven pharmacological therapies to help manage this condition, nutritional interventions are a safer option to help prevent and manage dyslipidemia. Addition of almonds in the daily diet has been proposed to beneficially impact the lipid profile. This review critically examines the available evidence assessing the effect of almonds on dyslipidemia in the South Asian (particularly Indian) context. An extensive review comprised of epidemiological studies, clinical trials, meta-analyses, and systematic reviews was conducted from published literature from across the world. Studies examining the effect of almonds on different aspects of dyslipidemia viz. high low-density lipoprotein-cholesterol (LDL-C), low high-density lipoprotein-cholesterol (HDL-C), triglyceridaemia, and high total cholesterol levels have been included. In several studies, almonds have been shown to reduce LDL-C-which is a known risk factor for CHD-and the effect of almonds has been well documented in systematic reviews and meta-analysis of clinical trials. Addition of almonds in the diet has been shown to not only to reduce LDL-C levels, but also to maintain HDL-C levels. This review provides information about the use of this simple nutritional strategy which may help manage known major risk factors for heart disease, such as high LDL-C and low HDL-C levels especially in the context of South Asians.


18F-Choline PET/MR Can Detect and Delineate Local Recurrence After High-Intensity Focused Ultrasound Therapy of Prostate Cancer.
Restaging local recurrence after high-intensity focused ultra-sound (HIFU) is based on multiparametric MRI (mpMRI). However, postinterventional changes of the tissue, such as edema or hemorrhage, are limiting tumor detection on mpMRI. We present a case of a rising prostate-specific antigen values, negative mpMRI, and a Gleason score of 4+4 on template biopsy after HIFU. On F-choline PET/MR, high focal uptake was detected at the location of positive biopsy. Re-HIFU based on the fused F-choline PET/MR images was performed, followed by a recurrence-free period of 11 months. Thus, F-choline PET/MR could improve guiding retreatment in patients with recurrence after HIFU.


Infinite Dilution Activity Coefficients as Constraints for Force Field Parametrization and Method Development.
Molecular simulations begin with an underlying energy model or force field and from this can predict diverse physical properties. However, force fields were often developed with relatively limited data sets, yet accuracy for diverse properties across a broad chemical space is desirable; therefore, tests of such accuracy are particularly important. Here, to this end, we calculated 237 infinite dilution activity coefficients (IDACs), comparing with experimental values from NIST's ThermoML database. We found that calculated IDAC values correlate strongly with experiment (Pearson R of 0.92 ± 0.01) and allow us to identify specific functional groups that appear to present challenges to the force field employed. One potentially valuable aspect of IDACs, as compared to solvation free energies, which have been frequently employed as force field tests, is that the same molecules serve both as solutes and solvents in different cases, allowing us to ensure that force fields are not overly tuned to one particular environment or solvent.


A synthesis of growth rates in marine epipelagic invertebrate zooplankton.
We present the most extensive study to date of globally compiled and analysed weight-specific growth rates in marine epi-pelagic invertebrate metazoan zooplankton. Using specified selection criteria, we analyse growth rates from a variety of zooplanktonic taxa, including both holo- and mero-planktonic forms, from over 110 published studies. Nine principal taxonomic groups are considered, the copepods (number of individual data points (n) = 2,528); crustaceans other than copepods (n = 253); cnidarians (n = 77); ctenophores (n = 27); chaetognaths (n = 87); pteropods (n = 8); polychaetes (n = 12); thaliaceans (n = 88); and larvaceans (n = 91). The copepods are further examined by subdividing them into broadcasters or sac-spawning species, and as nauplii (N1-N6), copepodites (C1-C5) and adults (C6). For each taxonomic group relationships between growth, temperature and body weight are examined using a variety of methods. Weight-specific growth tends to increase with increasing temperature and with decreasing body weight in the crustacean group. Growth does not relate to body weight in the case of chaetognaths and larvaceans, but does increase with temperature. In the cnidarian and ctenophore groups growth does not relate to temperature, but is negatively related to body size. For the thaliceans growth increases with both increasing body weight and temperature. In the entire broadcasting copepod data set, weight-specific growth increases with increasing temperature and decreasing body weight. In sac-spawners, growth increases with increasing temperature, and increases with decreasing body weight at temperatures below 20 degrees C, but decreases with body weight at temperatures above this. Comparison between the different taxa shows important differences and similarities. Our extensive synthesis of data generally confirms that larvaceans, pteropods, cnidarians and ctenophores have rates of weight-specific growth that are typically greater than the copepods, chaetognaths and other crustaceans of similar carbon weight. For the cnidarians, ctenophores and larvaceans growth rates are almost always greater than the general relationship describing copepod growth, and are also at the upper limits or beyond the maximum rates for copepods of a similar weight. For the pteropods, growth rates are generally greater than those of copepods, although the data set was limited to a single carnivorous species in a single study (i.e. Clione limacina). The thaliaceans have the highest growth rates for animals with body weights greater than around 1 mg C ind-1, with rates of up to 2.1 d-1 for Pegea bicaudata. Whilst the larvaceans can achieve rates of 2 d-1 in warm tropical waters (28 degrees C), and as high as > 3 d-1 for < 0.2 mg C individual-1 animals of Oikopleura diocia. These are possibly the highest rates every recorded in epi-pelagic metazoans. Reasons for the differences between taxonomic groups are discussed in relation to intrinsic and extrinsic factors and limitations. The importance of this investigation not only lies in it being the most comprehensive overview of patterns of growth to date, but because the data set highlight the gaps in measurements and current knowledge. We examine the inadequacies in the current data sets, and in the methods being used to measure growth and production. Most of the data are for animals collected from coastal and estuarine waters, and it is clear that for a fuller understanding there is an urgent need for work in the open ocean, and for investigations outside temperate regions. There is also a need to explore the role of food availability, and how food concentrations in incubations, and under food saturation, relate to those experienced in the natural environment.


Effect of arm cycling and task-oriented exercises on fatigue and upper limb performance in multiple sclerosis: a randomized crossover study.
Rehabilitation treatments have been proven to be a viable way to reduce fatigue and upper limb impairments in people with multiple sclerosis (PwMS). Our aim was to examine which treatment has better short-term and carryover effects on fatigue and manual dexterity in multiple sclerosis population. Twenty PwMS participated in a 16-week randomized crossover study composed of 20 sessions. The participants were divided into two groups (group A and group B). Sessions containing combined arm cycling and task-oriented exercises were administered by a physical therapist in hospital setting. Each group received 20 sessions of aerobic training and task-oriented exercises and then an 8-week rest period or vice versa with group A receiving sessions first. Fatigue was assessed by using the Modified Fatigue Impact Scale (MFIS) and Motor Fatigability Index (MFI), which was assessed using an engineered glove during a fatiguing finger tapping task. To measure manual dexterity, the nine hole peg test (NHPT) and a rate of tapping at maximum velocity task (RATE-MV) were utilized. Treatment effects were assessed by t-test or Mann-Whitney test at the end of both periods checking for carryover effects. After treatment the combined (Groups A and B) between-period differences were MFIS: 5.2 (10.7) points, P = 0.05; MFI: -0.007 (<0.001)Hz/s, P = 0.05 and RATE-MV: 0.2 (0.4) Hz/s, P = 0.05 in favor of the treatment period. No statistically significant between-period differences were found for the NHPT: 3.6 (25.0) s, P = 0.63. No carryover effects (P > 0.05) were observed. In conclusion, sessions of arm cycling and tailored task-oriented exercises have shown to be a viable resource for treating manual dexterity and fatigue in PwMS.


Child and adolescent mental health in schools.
It is a pleasure to write the editorial for this issue of the journal, which also highlights the publication of a Virtual Issue (VI)1 which brings together some of the most influential articles published on child and adolescent mental health in schools over the last 10 years. The retrospective by David Galloway, written 2001, is an interesting starting point for commentary, looking back over what is now 30 years. Its themes about UK educational policies and their impacts resonate through the articles collected here from across the world and are in most cases are as relevant today as ever (Galloway, 2001*)2 .


Synthesis of Biotin-Modified Galactosylated Chitosan Nanoparticles and Their Characteristics in Vitro and in Vivo.
Our previous study found that a nanoparticle drug delivery system that operates as a drug carrier and controlled release system not only improves the efficacy of the drugs but also reduces their side effects. However, this system could not efficiently target hepatoma cells. The aim of this study was to synthesize biotin-modified galactosylated chitosan nanoparticles (Bio-GC) and evaluate their characteristics in vitro and in vivo. Bio-GC nanomaterials were synthesized, and confirmed by fourier transform infrared spectroscopy (FT-IR) and hydrogen-1 nuclear magnetic resonance (1H-NMR). The liver position and cancer target property of Bio-GC nanoparticles in vitro and in vivo was tested by confocal laser and small animal imaging system. The characteristics of Bio-GC/5-fluorouracil (5-FU) nanoparticles in vitro and in vivo were explored by cell proliferation, migration and cytotoxicity test, or by animal experiment. Bio-GC nanoparticles were synthesized with biodegradable chitosan as the nanomaterial skeleton with biotin and galactose grafts. Bio-GC was confirmed by FT-IR and 1H-NMR. Bio-GC/5-FU nanoparticles were synthesized according to the optimal mass ratio for Bio-GC/5-FU (1: 4) and had a mean particle size of 81.1 nm, zeta potential of +39.2 mV, and drug loading capacity of 8.98%. Bio-GC/5-FU nanoparticles had sustained release properties (rapid, steady, and slow release phases). Bio-GC nanoparticles targeted liver and liver cancer cell in vitro and in vivo, and this was confirmed by confocal laser scanning and small animal imaging system. Compared with GC/5-FU nanoparticles, Bio-GC/5-FU nanoparticles showed more specific cytotoxic activity in a dose- and time-dependent manner and a more obvious inhibitory effect on the migration of liver cancer cells. In addition, Bio-GC/5-FU nanoparticles significantly prolonged the survival time of mice in orthotopic liver cancer transplantation model compared with other 5-FU nanoparticles or 5-FU alone. Bio-GC (0.64%) nanomaterial had no obvious cytotoxic effects on cells; thus, the concentration of Bio-GC/5-FU nanoparticles used was only 0.04% and showed no toxic effects on the cells. Bio-GC is a liver- and cancer-targeting nanomaterial. Bio-GC/5-FU nanoparticles as drug carriers have stronger inhibitory effects on the proliferation and migration of liver cancer cells compared with 5-FU in vitro and in vivo.


Elevated maternal serum-free β-human chorionic gonadotropin (β-hCG) and reduced risk of spontaneous preterm delivery.
Objective: To evaluate the relationship between first and second trimester maternal serum-free β-hCG and the risk of spontaneous preterm delivery (PTD). Study design: This was a case-control study of women evaluated and delivered at our institution from 2011 to 2015. Spontaneous PTD was defined as delivery before 37 weeks due to spontaneous preterm labor or premature rupture of membranes. Patient with multifetal gestation and those with medically indicated term or PTD were excluded. Results: Of 877 women meeting the inclusion criteria, 173 delivered preterm and 704 delivered at term, and 8.1% had high free β-hCG in one or both trimesters. High maternal first and/or second trimester free β-hCG (≥95th percentile) was associated with lower rates of PTD. Thirty-two women with high free β-hCG in both first and second trimesters delivered at term. Gestational age at delivery and birth weights were lower in women who did not have high free β-hCG in any trimester. Low free β-hCG (≤5th percentile) in either trimester was not associated with an increased or decreased likelihood of PTD. Logistic regression demonstrated an independent association of high free β-hCG (≥95th percentile) with a reduced likelihood of PTD. Stratified analysis revealed a stronger impact of this association in women with no prior history of PTD. Conclusions: High free β-hCG, in the absence of risk factors for medically indicated PTD, is associated with a reduced likelihood of spontaneous PTD and may represent a marker indicating lower risk.


Self-assembly of nucleocapsid-like particles from recombinant hepatitis C virus core protein.
Little is known about the assembly pathway and structure of hepatitis C virus (HCV) since insufficient quantities of purified virus are available for detailed biophysical and structural studies. Here, we show that bacterially expressed HCV core proteins can efficiently self-assemble in vitro into nucleocapsid-like particles. These particles have a regular, spherical morphology with a modal distribution of diameters of approximately 60 nm. Self-assembly of nucleocapsid-like particles requires structured RNA molecules. The 124 N-terminal residues of the core protein are sufficient for self-assembly into nucleocapsid-like particles. Inclusion of the carboxy-terminal domain of the core protein modifies the core assembly pathway such that the resultant particles have an irregular outline. However, these particles are similar in size and shape to those assembled from the 124 N-terminal residues of the core protein. These results provide novel opportunities to delineate protein-protein and protein-RNA interactions critical for HCV assembly, to study the molecular details of HCV assembly, and for performing high-throughput screening of assembly inhibitors.


Plant Defense Response to Fungal Pathogens (II. G-Protein-Mediated Changes in Host Plasma Membrane Redox Reactions).
Elicitor preparations containing the avr5 gene products from races 4 and 2.3 of Cladosporium fulvum, and tomato (Lycopersicon esculentum L.) cells containing the resistance gene Cf5 were used to investigate the involvement of redox processes in the production of active oxygen species associated with the plant response to the fungal elicitors. Here we demonstrate that certain race-specific elicitors of C. fulvum induced an increase in ferricyanide reduction in enriched plasma membrane fractions of tomato cells. The addition of elicitors to plasma membranes also induced increases in NADH oxidase and NADH-dependent cytochrome c reductase activities, whereas ascorbate peroxidase activity was decreased. These results suggest that changes in the host plasma membrane redox processes, transferring electrons from reducing agents to oxygen, could be involved in the increased production of active oxygen species by the race-specific elicitors. Our results also show that the dephosphorylation of enzymes involved in redox reactions is responsible for the race-specific induced redox activity. The effects of guanidine nucleotide analogs and mastoparan on the activation of plasma membrane redox reactions support the role of GTP-binding proteins in the transduction of signals leading to the activation of the defense response mechanisms of tomato against fungal pathogens.


On-demand acoustic droplet splitting and steering in a disposable microfluidic chip.
On-chip droplet splitting is one of the fundamental droplet-based microfluidic unit operations to control droplet volume after production and increase operational capability, flexibility, and throughput. Various droplet splitting methods have been proposed, and among them the acoustic droplet splitting method is promising because of its label-free operation without any physical or thermal damage to droplets. Previous acoustic droplet splitting methods faced several limitations: first, they employed a cross-type acoustofluidic device that precluded multichannel droplet splitting; second, they required irreversible bonding between a piezoelectric substrate and a microfluidic chip, such that the fluidic chip was not replaceable. Here, we present a parallel-type acoustofluidic device with a disposable microfluidic chip to address the limitations of previous acoustic droplet splitting devices. In the proposed device, an acoustic field is applied in the direction opposite to the flow direction to achieve multichannel droplet splitting and steering. A disposable polydimethylsiloxane microfluidic chip is employed in the developed device, thereby removing the need for permanent bonding and improving the flexibility of the droplet microfluidic device. We experimentally demonstrated on-demand acoustic droplet bi-splitting and steering with precise control over the droplet splitting ratio, and we investigated the underlying physical mechanisms of droplet splitting and steering based on Laplace pressure and ray acoustics analyses, respectively. We also demonstrated droplet tri-splitting to prove the feasibility of multichannel droplet splitting. The proposed on-demand acoustic droplet splitting device enables on-chip droplet volume control in various droplet-based microfluidic applications.


The effectiveness of healthy meals at work on reaction time, mood and dietary intake: a randomised cross-over study in daytime and shift workers at an university hospital.
Our dietary habits affect both cognitive performance and mood. The aim of the study was to examine the effect of increased availability of healthy meals and water at work on healthcare staff. The study used an 8-week randomised cross-over design. A total of sixty physicians, nurses and nursing assistants, including sixteen working on shifts, were recruited. The participants received a self-selected keyhole-labelled (Nordic nutrition label) lunch, snack and bottled water during each shift throughout the intervention period. Reaction time (Go/No-Go test), mood-related scores (POMS) and dietary intake were assessed at run-in, and at the end of the intervention and the control periods. The intake of fat (P=0·030) and PUFA (P=0·003) was lower, and the intake of carbohydrate (P=0·008), dietary fibre (P=0·031) and water (P<0·001) was greater in the intervention period than in the control period. The intervention had no effect on reaction time or any of the mood-related scores in the group as a whole. In shift-working participants, the intervention period resulted in a 31·1 % lower Fatigue-Inertia Score (P=0·003), a 15·3 % higher Vigour-Activity Score (P=0·041) and a 42·7 % lower Total Mood Disturbance Score (P=0·017), whereas the only dietary component that significantly improved was water intake (P=0·034), when compared with the control period. Providing healthy meals, snacks and water during working hours seems to be an effective way of improving employees' dietary intake. Moreover, increased intake of water may be associated with beneficial effects on fatigue, vigour and total mood in shift-working healthcare staff.


Pelvic fracture hemorrhage. Priorities in management.
Hemorrhage remains the leading cause of mortality in patients with severe pelvic fractures. To evaluate diagnostic and treatment priorities for this problem, we retrospectively reviewed 245 consecutive patients admitted to our institution with pelvic fractures. Supraumbilical diagnostic peritoneal lavage (DPL) was grossly positive in 27 patients, and eight (30%) of these had life-threatening intra-abdominal hemorrhage identified at laparotomy. No patient with a positive DPL by count alone had life-threatening intra-abdominal hemorrhage. Pelvic fracture stabilization with early external pelvic fixation was associated with less requirement for blood transfusion (10 +/- 1 U) than with the pneumatic antishock garment (17 +/- 3 U). Nine patients with pelvic arterial injuries underwent angiographic embolization, and eight patients died (89%). We conclude that pelvic angiography should be performed before laparotomy in hemodynamically unstable patients with pelvic fracture, unless the DPL is grossly positive.


The effects of enactment on communicative competence in aphasic casual conversation: a functional linguistic perspective.
Previous research has shown that speakers with aphasia rely on enactment more often than non-brain-damaged language users. Several studies have been conducted to explain this observed increase, demonstrating that spoken language containing enactment is easier to produce and is more engaging to the conversation partner. This paper describes the effects of the occurrence of enactment in casual conversation involving individuals with aphasia on its level of conversational assertiveness. To evaluate whether and to what extent the occurrence of enactment in speech of individuals with aphasia contributes to its conversational assertiveness. Conversations between a speaker with aphasia and his wife (drawn from AphasiaBank) were analysed in several steps. First, the transcripts were divided into moves, and all moves were coded according to the systemic functional linguistics (SFL) framework. Next, all moves were labelled in terms of their level of conversational assertiveness, as defined in the previous literature. Finally, all enactments were identified and their level of conversational assertiveness was compared with that of non-enactments. Throughout their conversations, the non-brain-damaged speaker was more assertive than the speaker with aphasia. However, the speaker with aphasia produced more enactments than the non-brain-damaged speaker. The moves of the speaker with aphasia containing enactment were more assertive than those without enactment. The use of enactment in the conversations under study positively affected the level of conversational assertiveness of the speaker with aphasia, a competence that is important for speakers with aphasia because it contributes to their floor time, chances to be heard seriously and degree of control over the conversation topic.


A predictive model for delirium in hospitalized elderly medical patients based on admission characteristics.
To prospectively develop and validate a predictive model for the occurrence of new delirium in hospitalized elderly medical patients based on characteristics present at admission. Two prospective cohort studies done in tandem. University teaching hospital. The development cohort included 107 hospitalized general medical patients 70 years or older who did not have dementia or delirium at admission. The validation cohort included 174 comparable patients. Patients were assessed daily for delirium using a standardized, validated instrument. The predictive model developed in the initial cohort was then validated in a separate cohort of patients. Delirium developed in 27 of 107 patients (25%) in the development cohort. Four independent baseline risk factors for delirium were identified using proportional hazards analysis: These included vision impairment (adjusted relative risk, 3.5; 95% Cl, 1.2 to 10.7); severe illness (relative risk, 3.5; Cl, 1.5 to 8.2); cognitive impairment (relative risk, 2.8; Cl, 1.2 to 6.7); and a high blood urea nitrogen/creatinine ratio (relative risk, 2.0; Cl, 0.9 to 4.6). A risk stratification system was developed by assigning 1 point for each risk factor present. Rates of delirium for low- (0 points), intermediate- (1 to 2 points), and high-risk (3 to 4 points) groups were 9%, 23%, and 83% (P < 0.0001), respectively. The corresponding rates in the validation cohort, in which 29 of 174 patients (17%) developed delirium, were 3%, 16%, and 32% (P < 0.002). The rates of death or nursing home placement, outcomes potentially related to delirium, were 9%, 16%, and 42% (P = 0.02) in the development cohort and 3%, 14%, and 26% (P = 0.007) in the validation cohort. Delirium among elderly hospitalized patients is common, and a simple predictive model based on four risk factors can be used at admission to identify elderly persons at the greatest risk.


Altering the Speed Profiles of Wheelchair Rugby Players With Game-Simulation Drill Design.
To examine the speed profiles of elite wheelchair rugby (WCR) players during game-simulation training drills of differing player number and shot-clock regulations. A secondary aim was to determine whether the profiles were further influenced by player classification. Eight elite WCR players (low-point n = 3, high-point n = 5) were monitored using a radio-frequency-based indoor tracking system during training sessions over a 5-mo period. Speed profiles were collected for 3 modified game-simulation drills-3-versus-3 drills (n = 8 observations), 30-s shot clock (n = 24 observations), and 15-s shot clock (n = 16 observations)-and were compared with regular game-simulation drills (4 vs 4, 40-s shot clock; n = 16 observations). Measures included mean and peak speed; exercise-intensity ratios, defined as the ratio of time spent performing at high and low speeds; and the number of high-speed activities performed. Compared with regular game-simulation drills, 3-versus-3 drills elicited a moderate increase in mean speed (6.3%; effect size [ES] = 0.7) and the number of high-speed activities performed (44.1%; ES = 1.1). Minimal changes in speed profiles were observed during the 30-s shot clock, although moderate to large increases in all measures were observed during the 15-s shot-clock drills. Classification-specific differences were further identified, with increased activity observed for high-point players during the 3-versus-3 drill and for low-point players during the 15-s shot clock. By reducing the number of players on court and the shot clock to 15 s, coaches can significantly increase elite WCR players' speed profiles during game-simulation drills.


Out of the laboratory, into the field: perspectives on social, ethical and regulatory challenges in UK wildlife research.
Drawing on insights from qualitative social science research, this paper aims to prompt reflection on social, ethical and regulatory challenges faced by scientists undertaking invasive animal research in the field and propose ways of addressing these challenges to promote good care for animals and environments. In particular, we explore challenges relating to the management of (i) relationships with publics and stakeholders, who may be present at field sites or crucial to research success; (ii) ethical considerations not present in the laboratory, such as the impacts of research on populations and ecosystems; (iii) working under an array of regulations, which may operate in accordance with competing ethical principles or objectives; and (iv) relationships with regulators (especially vets), which may involve disagreements over ethics and expertise, especially because regulators may be more accustomed to overseeing research in the laboratory than the field. We argue that flexibility-at a personal and policy level-and respect for others' expertise emerged as two key ways of negotiating ethical challenges, fostering positive working relationships and promoting good care for individual animals and broader ecosystems. While our analysis focuses on the UK, we propose that many of these lessons are broadly applicable to international contexts. This article is part of the theme issue 'Measuring physiology in free-living animals (Part II)'.


BCG-induced protection against Mycobacterium tuberculosis infection: Evidence, mechanisms, and implications for next-generation vaccines.
The tuberculosis (TB) vaccine Bacillus Calmette-Guérin (BCG) was introduced 100 years ago, but as it provides insufficient protection against TB disease, especially in adults, new vaccines are being developed and evaluated. The discovery that BCG protects humans from becoming infected with Mycobacterium tuberculosis (Mtb) and not just from progressing to TB disease provides justification for considering Mtb infection as an endpoint in vaccine trials. Such trials would require fewer participants than those with disease as an endpoint. In this review, we first define Mtb infection and disease phenotypes that can be used for mechanistic studies and/or endpoints for vaccine trials. Secondly, we review the evidence for BCG-induced protection against Mtb infection from observational and BCG re-vaccination studies, and discuss limitations and variation of this protection. Thirdly, we review possible underlying mechanisms for BCG efficacy against Mtb infection, including alternative T cell responses, antibody-mediated protection, and innate immune mechanisms, with a specific focus on BCG-induced trained immunity, which involves epigenetic and metabolic reprogramming of innate immune cells. Finally, we discuss the implications for further studies of BCG efficacy against Mtb infection, including for mechanistic research, and their relevance to the design and evaluation of new TB vaccines.


PPE37 Is Essential for Mycobacterium tuberculosis Heme-Iron Acquisition (HIA), and a Defective PPE37 in Mycobacterium bovis BCG Prevents HIA.
Mycobacterium tuberculosis, one of the world's leading causes of death, must acquire nutrients, such as iron, from the host to multiply and cause disease. Iron is an essential metal and M. tuberculosis possesses two different systems to acquire iron from its environment: siderophore-mediated iron acquisition (SMIA) and heme-iron acquisition (HIA), involving uptake and degradation of heme to release ferrous iron. We have discovered that Mycobacterium bovis BCG, the tuberculosis vaccine strain, is severely deficient in HIA, and we exploited this phenotypic difference between BCG and M. tuberculosis to identify genes involved in HIA by complementing BCG's defect with a fosmid library. We identified ppe37, an iron-regulated PPE family gene, as being essential for HIA. BCG complemented with M. tuberculosisppe37 exhibits HIA as efficient as that of M. tuberculosis, achieving robust growth with <0.2 µM hemin. Conversely, deletion of ppe37 from M. tuberculosis results in a strain severely attenuated in HIA, with a phenotype nearly identical to that of BCG, requiring a 200-fold higher concentration of hemin to achieve growth equivalent to that of its parental strain. A nine-amino-acid deletion near the N terminus of BCG PPE37 (amino acids 31 to 39 of the M. tuberculosis PPE37 protein) underlies BCG's profound defect in HIA. Significant genetic variability exists in ppe37 genes across different M. tuberculosis strains, with more than 60% of sequences from completely sequenced M. tuberculosis genomes having mutations that result in altered PPE37 proteins; furthermore, these altered PPE37 proteins are nonfunctional in HIA. Our findings should allow delineation of the relative roles of HIA and SMIA in M. tuberculosis pathogenesis.


Selective Sweeps and Parallel Pathoadaptation Drive Pseudomonas aeruginosa Evolution in the Cystic Fibrosis Lung.
Pulmonary infections caused by Pseudomonas aeruginosa are a recalcitrant problem in cystic fibrosis (CF) patients. While the clinical implications and long-term evolutionary patterns of these infections are well studied, we know little about the short-term population dynamics that enable this pathogen to persist despite aggressive antimicrobial therapy. Here, we describe a short-term population genomic analysis of 233 P. aeruginosa isolates collected from 12 sputum specimens obtained over a 1-year period from a single patient. Whole-genome sequencing and antimicrobial susceptibility profiling identified the expansion of two clonal lineages. The first lineage originated from the coalescence of the entire sample less than 3 years before the end of the study and gave rise to a high-diversity ancestral population. The second expansion occurred 2 years later and gave rise to a derived population with a strong signal of positive selection. These events show characteristics consistent with recurrent selective sweeps. While we cannot identify the specific mutations responsible for the origins of the clonal lineages, we find that the majority of mutations occur in loci previously associated with virulence and resistance. Additionally, approximately one-third of all mutations occur in loci that are mutated multiple times, highlighting the importance of parallel pathoadaptation. One such locus is the gene encoding penicillin-binding protein 3, which received three independent mutations. Our functional analysis of these alleles shows that they provide differential fitness benefits dependent on the antibiotic under selection. These data reveal that bacterial populations can undergo extensive and dramatic changes that are not revealed by lower-resolution analyses. Pseudomonas aeruginosa is a bacterial opportunistic pathogen responsible for significant morbidity and mortality in cystic fibrosis (CF) patients. Once it has colonized the lung in CF, it is highly resilient and rarely eradicated. This study presents a deep sampling examination of the fine-scale evolutionary dynamics of P. aeruginosa in the lungs of a chronically infected CF patient. We show that diversity of P. aeruginosa is driven by recurrent clonal emergence and expansion within this patient and identify potential adaptive variants associated with these events. This high-resolution sequencing strategy thus reveals important intraspecies dynamics that explain a clinically important phenomenon not evident at a lower-resolution analysis of community structure.


Epidemiological, social, diagnostic and economic evaluation of population screening for genital chlamydial infection.
To investigate epidemiological, social, diagnostic and economic aspects of chlamydia screening in non-genitourinary medicine settings. Linked studies around a cross-sectional population-based survey of adult men and women invited to collect urine and (for women) vulvovaginal swab specimens at home and mail these to a laboratory for testing for Chlamydia trachomatis. Specimens were used in laboratory evaluations of an amplified enzyme immunoassay (PCE EIA) and two nucleic acid amplification tests [Cobas polymerase chain reaction (PCR), Becton Dickinson strand displacement amplification (SDA)]. Chlamydia-positive cases and two negative controls completed a risk factor questionnaire. Chlamydia-positive cases were invited into a randomised controlled trial of partner notification strategies. Samples of individuals testing negative completed psychological questionnaires before and after screening. In-depth interviews were conducted at all stages of screening. Chlamydia transmission and cost-effectiveness of screening were investigated in a transmission dynamic model. General population in the Bristol and Birmingham areas of England. In total, 19,773 women and men aged 16-39 years were randomly selected from 27 general practice lists. Screening invitations reached 73% (14,382/19,773). Uptake (4731 participants), weighted for sampling, was 39.5% (95% CI 37.7, 40.8%) in women and 29.5% (95% CI 28.0, 31.0%) in men aged 16-39 years. Chlamydia prevalence (219 positive results) in 16-24 year olds was 6.2% (95% CI 4.9, 7.8%) in women and 5.3% (95% CI 4.4, 6.3%) in men. The case-control study did not identify any additional factors that would help target screening. Screening did not adversely affect anxiety, depression or self-esteem. Participants welcomed the convenience and privacy of home-sampling. The relative sensitivity of PCR on male urine specimens was 100% (95% CI 89.1, 100%). The combined relative sensitivities of PCR and SDA using female urine and vulvovaginal swabs were 91.8% (86.1, 95.7, 134/146) and 97.3% (93.1, 99.2%, 142/146). A total of 140 people (74% of eligible) participated in the randomised trial. Compared with referral to a genitourinary medicine clinic, partner notification by practice nurses resulted in 12.4% (95% CI -3.7, 28.6%) more patients with at least one partner treated and 22.0% (95% CI 6.1, 37.8%) more patients with all partners treated. The health service and patients costs (2005 prices) of home-based postal chlamydia screening were 21.47 pounds (95% CI 19.91 pounds, 25.99) per screening invitation and 28.56 pounds (95% CI 22.10 pounds, 30.43) per accepted offer. Preliminary modelling found an incremental cost-effectiveness ratio (2003 prices) comparing screening men and women annually to no screening in the base case of 27,000 pounds/major outcome averted at 8 years. If estimated screening uptake and pelvic inflammatory disease incidence were increased, the cost-effectiveness ratio fell to 3700 pounds/major outcome averted. Proactive screening for chlamydia in women and men using home-collected specimens was feasible and acceptable. Chlamydia prevalence rates in men and women in the general population are similar. Nucleic acid amplification tests can be used on first-catch urine specimens and vulvovaginal swabs. The administrative costs of proactive screening were similar to those for opportunistic screening. Using empirical estimates of screening uptake and incidence of complications, screening was not cost-effective.


Long-term results following osteotomy of the thumb delta phalanx in Rubinstein-Taybi Syndrome.
Rubinstein-Taybi Syndrome is a rare condition affecting 1:125,000 children. It is associated with short broad radially deviated thumbs, secondary to a delta proximal phalanx of the thumb. We undertook a retrospective review of seven children (13 thumbs) with Rubinstein-Taybi syndrome whose thumbs were treated using a corrective osteotomy to the delta phalanx over a 13 year period. The types of osteotomy used in the series were reverse wedge osteotomy, opening wedge osteotomy and dome shaped osteotomy. The mean preoperative radial deviation of thumbs was 68 degrees (range 45-85 degrees ). At follow up five of the 13 thumbs demonstrated some residual radial deviation. All recurrences occurred in the dome shaped osteotomy group. Our data suggest that surgery is effective in correcting the deformity, but there is a risk of incomplete correction or recurrence. Despite the recurrence the mean postoperative deformity was significantly better than preoperatively and the majority of patients families subjectively reported good function. No patient in our series has yet undergone further corrective surgery.


The impact of the first COVID-19 lockdown in the UK for doctoral and early career researchers.
Doctoral researchers and early career researchers (ECRs) are crucial to producing scientific advancements and represent the future of academic leadership. Their research endeavours were changed radically by lockdowns in response to the COVID-19 pandemic. The aim of this study was to explore the perceived benefits and challenges of the national lockdown in the UK from the perspective of doctoral researchers and ECRs. We present analysis of qualitative survey data from 1,142 doctoral researchers and ECRs on their experiences of the first UK lockdown collected from April 16, 2020-May 14, 2020. Our findings suggest considerable heterogeneity in how the pandemic impacted this key group of academic workers. Challenges arising from the lockdown largely cohered around a poor work environment, limited access to resources, perceptions of pressure, and negative psychological outcomes. Conversely, respondents also highlighted several benefits in the early stages of the pandemic, with the change to working from home creating more time, resulting in greater productivity and a better work-life balance. Collectively, findings indicate the importance of considering the personal circumstances and needs of individual researchers. We discuss the implications for support these researchers require to rebuild their careers in the wake of the initial disruption.


Opportunities at the intersection of bioinformatics and health informatics: a case study.
This paper provides a "viewpoint discussion" based on a presentation made to the 2000 Symposium of the American College of Medical Informatics. It discusses potential opportunities for researchers in health informatics to become involved in the rapidly growing field of bioinformatics, using the activities of the Yale Center for Medical Informatics as a case study. One set of opportunities occurs where bioinformatics research itself intersects with the clinical world. Examples include the correlations between individual genetic variation with clinical risk factors, disease presentation, and differential response to treatment; and the implications of including genetic test results in the patient record, which raises clinical decision support issues as well as legal and ethical issues. A second set of opportunities occurs where bioinformatics research can benefit from the technologic expertise and approaches that informaticians have used extensively in the clinical arena. Examples include database organization and knowledge representation, data mining, and modeling and simulation. Microarray technology is discussed as a specific potential area for collaboration. Related questions concern how best to establish collaborations with bioscientists so that the interests and needs of both sets of researchers can be met in a synergistic fashion, and the most appropriate home for bioinformatics in an academic medical center.


Human intestinal development in a severe-combined immunodeficient xenograft model.
The present work describes a severe-combined immunodeficient murine xenograft model used to investigate human gastrointestinal ontogenesis. Specifically, the study has tested whether carefully selected regions of human fetal gut are able to undergo region-specific morphogenesis and epithelial cytodifferentiation when transplanted subcutaneously into immunodeficient mice. In addition, double-label in situ hybridisation techniques, utilising specific human and mouse DNA probes, have been adopted to characterise host and donor cell types and to investigate the potential developmental roles for non-epithelial cells in the regulation of epithelial differentiation pathways in vivo. Human fetal small and large bowel developed to form a characteristic mucosa 10 weeks after transplantation, which displayed clear region-specific structural and functional gradients. The initial phase of xenograft epithelialisation closely resembled the stratified type of epithelium which is present during early fetal gastrointestinal development. Idiosyncratic epithelial differentiation pathways were recorded during xenograft regeneration, with an absence of Paneth cells and an abundance of enteroendocrine cells when compared with developed xenograft and paediatric intestine. Such differences may, therefore, be important in ensuring rapid and region-specific development in the absence of conventional luminal stimuli and hormonal changes that occur normally during pregnancy. In situ hybridisation demonstrated an exclusively human origin for the intestinal xenograft epithelium and muscularis mucosa and externa. Although the submucosa and lamina propria were comprised of a chimeric mixture, murine cells were rarely seen to contact with the epithelium, which interacted primarily with human myofibroblasts and human intraepithelial lymphocytes. It is proposed that a 'selection' process operates to maintain species-specific cellular interactions, and this mechanism may subsequently play an important role in regulating epithelial cell differentiation, orchestrated in part by juxtaposed non-epithelial cell types.


DUOX2 variants associate with preclinical disturbances in microbiota-immune homeostasis and increased inflammatory bowel disease risk.
A primordial gut-epithelial innate defense response is the release of hydrogen peroxide by dual NADPH oxidase (DUOX). In inflammatory bowel disease (IBD), a condition characterized by an imbalanced gut microbiota-immune homeostasis, DUOX2 isoenzyme is the highest induced gene. Performing multiomic analyses using 2872 human participants of a wellness program, we detected a substantial burden of rare protein-altering DUOX2 gene variants of unknown physiologic significance. We identified a significant association between these rare loss-of-function variants and increased plasma levels of interleukin-17C, which is induced also in mucosal biopsies of patients with IBD. DUOX2-deficient mice replicated increased IL-17C induction in the intestine, with outlier high Il17c expression linked to the mucosal expansion of specific Proteobacteria pathobionts. Integrated microbiota/host gene expression analyses in patients with IBD corroborated IL-17C as a marker for epithelial activation by gram-negative bacteria. Finally, the impact of DUOX2 variants on IL-17C induction provided a rationale for variant stratification in case control studies that substantiated DUOX2 as an IBD risk gene. Thus, our study identifies an association of deleterious DUOX2 variants with a preclinical hallmark of disturbed microbiota-immune homeostasis that appears to precede the manifestation of IBD.


Risk factors for and impact of carbapenem-resistant Acinetobacter baumannii colonization and infection: matched case-control study.
The objective of this investigation was to identify risk factors for carbapenem-resistant Acinetobacter baumannii (CRAB) and its association with mortality. A population-based matched case-control study using the computerized database of Clalit Health Services (CHS) in the period between 2007 and 2012 was conducted. Hospitalized patients with CRAB colonization or infection were compared to hospitalized patients without evidence of A. baumannii, matched by age, ward of hospitalization, season, Charlson score, and length of hospitalization. Risk factors for CRAB isolation were searched for using multivariate analysis. Association of CRAB and other risk factors with mortality were assessed in the cohort. A total of 1190 patients with CRAB were matched to 1190 patients without CRAB. Low socioeconomic status was independently associated with CRAB isolation and CRAB bacteremia [odds ratio 2.18, 95% confidence interval (CI) 1.02-5]. Other risk factors were invasive procedures and bacteremia with other pathogens prior to CRAB isolation, and various comorbidities. Among all patients, CRAB isolation was independently associated with increased mortality (hazard ratio 2.33, 95% CI 2.08-2.6). Socioeconomic status is associated with health outcomes. Our population-based study revealed an almost doubled risk for CRAB in patients at lower socioeconomic status and an association with healthcare exposure. CRAB was associated with mortality and might become a risk indicator for complex morbidity and mortality.


Stress-related and spontaneous stage differentiation of Toxoplasma gondii.
Toxoplasma gondii is an obligatory intracellular parasitic protozoan that infects a variety of avian and mammalian hosts including up to one third of the human population worldwide. Developmental differentiation between distinct stages, i.e. sporozoites, tachyzoites and bradyzoites is fundamental for the parasite life cycle and for transmission between hosts. It is also interconnected with the pathogenesis of overt toxoplasmosis and makes T. gondii an important opportunistic pathogen of humans. In order to delineate the underlying mechanisms, several cell culture differentiation systems have been developed which mimic the transition from fast-replicating tachyzoites to slowly proliferating bradyzoites in vitro. Since exogenous stress factors, i.e. alkaline pH, IFN-gamma and other proinflammatory cytokines, chemicals or drugs, heat shock, and deprivation of nutrients have been shown to increase the efficacy of bradyzoite development in vitro, Toxoplasma stage differentiation is largely viewed as a stress-related response to hostile environmental conditions. However, tachyzoite to bradyzoite differentiation also occurs spontaneously in vitro and this raises questions about the importance of stress conditions for triggering stage conversion. High frequencies of spontaneous bradyzoite development in primary and permanent skeletal muscle cells, i.e. cells that preferentially harbour bradyzoite-containing tissue cysts in vivo suggest that the host cell type may be critical. Furthermore, the host cell transcriptome, including the expression of distinct host cell genes, has recently been shown to trigger bradyzoite development and cyst formation. Together, these results strongly indicate that the complex cellular environment, besides exogenous stress factors, may govern the developmental differentiation of T. gondii.


Provision of pharmaceutical care by community pharmacists across Europe: Is it developing and spreading?
Pharmaceutical care involves patient-centred pharmacist activity to improve medicines management by patients. The implementation of this service in a comprehensive manner, however, requires considerable organisation and effort, and indeed, it is often not fully implemented in care settings. The main objective was to assess how pharmaceutical care provision within community pharmacy has evolved over time in Europe. A cross-sectional questionnaire-based survey of community pharmacies, using a modified version of the Behavioural Pharmaceutical Care Scale (BPCS) was conducted in late 2012/early 2013 within 16 European countries and compared with an earlier assessment conducted in 2006. The provision of comprehensive pharmaceutical care has slightly improved in all European countries that participated in both editions of this survey (n = 8) with progress being made particularly in Denmark and Switzerland. Moreover, there was a wider country uptake, indicating spread of the concept. However, due to a number of limitations, the results should be interpreted with caution. Using combined data from participating countries, the provision of pharmaceutical care was positively correlated with the participation of the community pharmacists in patient-centred activities, routine use of pharmacy software with access to clinical data, participation in multidisciplinary team meetings, and having specialized education. The present study demonstrated a slight evolution in self-reported provision of pharmaceutical care by community pharmacists across Europe, as measured by the BPCS. The slow progress suggests a range of barriers, which are preventing pharmacists moving beyond traditional roles. Support from professional bodies and more patient-centred community pharmacy contracts, including remuneration for pharmaceutical care services, are likely to be required if quicker progress is to be made in the future.


A paternally inherited duplication in the Prader-Willi/Angelman syndrome critical region: a case and family study.
The Prader-Willi/Angelman Critical Region (PWACR; Chromosome 15q11-13) is of interest as a potential locus for genes conferring susceptibility to autism spectrum disorders (ASD). This report describes a female proband referred for evaluation of a possible ASD. Genetic analyses indicated that the proband, her father and one of her sisters, carried a paternally derived interstitial duplication involving 15q11-13. The proband showed evidence of ASD (PDD-NOS), borderline mental retardation, mild hypotonia and joint laxity. Her father and her sister were of normal intelligence and neither was thought to have an ASD, although speech/language difficulties and some autistic type behaviours were reported to have been present early in the development of the sister. This is one of the first reports of a child with a paternal duplication and an autism spectrum disorder. More research is required to determine whether paternally derived duplications that involve 15q11-13 are associated with developmental impairments.


Height and its relationship to refraction and biometry parameters in Singapore Chinese children.
To examine the association between the anthropometric measurements of height, weight, body mass index (BMI) and refraction and other ocular parameters in Singapore Chinese children. In a cross-sectional study of 1449 Chinese schoolchildren, aged 7 to 9 years, from three Singapore schools, height and weight were measured according to standard protocol, and BMI was calculated. Refractive error and corneal curvature measures were determined by autorefraction in eyes under cycloplegia. Axial length, vitreous chamber depth, lens thickness, and anterior chamber depth were measured using A-scan biometry ultrasonography. In comparison with the children with height in the first quartile for a given age and gender, the eyeball length in children in the fourth quartile was 0.46 mm longer, the vitreous chamber depth 0.46 mm deeper, the corneal radius of curvature 0.10 mm greater (i.e., flatter), refraction more negative by 0.47 D (-0.76 D versus -0.29 D), and axial length-to-corneal curvature radius (AL-CR) ratio higher, after analyses controlling for age, gender, parental myopia, reading, school, and weight. The associations of height with refractive error and AL-CR ratio were significant in girls but not in boys. Heavier and more obese children had refractions that were more hyperopic (P = 0.01, P = 0.08), after analyses controlling for age, gender, parental myopia, reading, and school (height was also controlled for if weight was evaluated). This association was present in boys but not in girls. Controlling for age, gender, parental myopia, reading, school, and weight showed that taller Singapore Chinese children had eyes with longer axial lengths, deeper vitreous chambers, flatter corneas, and refractions that tended toward myopia. In multivariate analysis, eyes in children who were heavier or who had a higher BMI tended to have refractions that were more hyperopic, and eyes in heavier children had shorter vitreous chambers. Differences between the present results and a recent report in Singapore adults suggest either a cohort effect or a potential influence of systemic endocrine or metabolic factors during childhood on refractive development.


Prevalence of bacteremia in hospitalized pediatric patients with community-acquired pneumonia.
National guidelines recommend obtaining blood cultures in children hospitalized with moderate or severe community-acquired pneumonia (CAP). The objectives of this study were to determine the prevalence of bacteremia in children, identify factors associated with bacteremia and quantify the influence of positive blood cultures on clinical management in children hospitalized with CAP. This multicenter retrospective study included children from 60 days to 18 years of age requiring hospitalization for CAP. Categories analyzed were bacteremia, culture negative and no culture. Blood cultures were performed in 369 (56%) of 658 children with CAP. The prevalence of bacteremia was 7% (4.7-10.1%) in patients with a blood culture obtained. Bacteremia occurred in 21% of patients with a pleural drainage procedure and 75% of patients with distant site of infection (eg, osteomyelitis). Patients with bacteremia had longer duration of fever before admission and higher C-reactive protein values compared with those with negative or no blood culture. However, differences in white blood cell count and erythrocyte sedimentation rate between those with bacteremia and those without were not significant. Contamination rates were low and similar across institutions, ranging from 1% to 3.8% (P = 0.63). Blood culture-directed changes in antibiotic management occurred in 33% of patients with a contaminated culture and 65% of bacteremic patients. Antibiotic therapy was narrowed in 26% of bacteremic patients at hospital discharge. The prevalence of bacteremia was higher than previously reported in children hospitalized with CAP and consistent across children's hospitals. Positive blood cultures should prompt change to narrow-spectrum antibiotic therapy.


Benchmark of thirteen bioinformatic pipelines for metagenomic virus diagnostics using datasets from clinical samples.
Metagenomic sequencing is increasingly being used in clinical settings for difficult to diagnose cases. The performance of viral metagenomic protocols relies to a large extent on the bioinformatic analysis. In this study, the European Society for Clinical Virology (ESCV) Network on NGS (ENNGS) initiated a benchmark of metagenomic pipelines currently used in clinical virological laboratories. Metagenomic datasets from 13 clinical samples from patients with encephalitis or viral respiratory infections characterized by PCR were selected. The datasets were analyzed with 13 different pipelines currently used in virological diagnostic laboratories of participating ENNGS members. The pipelines and classification tools were: Centrifuge, DAMIAN, DIAMOND, DNASTAR, FEVIR, Genome Detective, Jovian, MetaMIC, MetaMix, One Codex, RIEMS, VirMet, and Taxonomer. Performance, characteristics, clinical use, and user-friendliness of these pipelines were analyzed. Overall, viral pathogens with high loads were detected by all the evaluated metagenomic pipelines. In contrast, lower abundance pathogens and mixed infections were only detected by 3/13 pipelines, namely DNASTAR, FEVIR, and MetaMix. Overall sensitivity ranged from 80% (10/13) to 100% (13/13 datasets). Overall positive predictive value ranged from 71-100%. The majority of the pipelines classified sequences based on nucleotide similarity (8/13), only a minority used amino acid similarity, and 6 of the 13 pipelines assembled sequences de novo. No clear differences in performance were detected that correlated with these classification approaches. Read counts of target viruses varied between the pipelines over a range of 2-3 log, indicating differences in limit of detection. A wide variety of viral metagenomic pipelines is currently used in the participating clinical diagnostic laboratories. Detection of low abundant viral pathogens and mixed infections remains a challenge, implicating the need for standardization and validation of metagenomic analysis for clinical diagnostic use. Future studies should address the selective effects due to the choice of different reference viral databases.


Genetic predictors associated with improvement of asthma symptoms in response to inhaled corticosteroids.
To date, genome-wide association studies (GWASs) of inhaled corticosteroid (ICS) response in asthmatic patients have focused primarily on lung function and exacerbations. We hypothesized that GWAS analysis could identify novel genetic markers predicting a symptomatic response to ICSs. We analyzed differences in asthma symptoms in response to ICSs in 124 white children from the Childhood Asthma Management Program (CAMP) trial using scores from diary cards. Of the 440,862 single nucleotide polymorphisms (SNPs) analyzed, the top 100 ranked SNPs were pursued for replication initially in subjects from the pediatric Childhood Asthma Research and Education trials (77 white children) and then in subjects from the adult Asthma Clinical Research Network (110 white adults) and Leukotriene Modifier or Corticosteroid or Corticosteroid-Salmeterol trials (110 white adults). The lowest P value for GWAS analysis in the CAMP trial was 8.94 × 10(-8) (rs2388639). Of the 60 SNPs available in the Childhood Asthma Research and Education Network trials, rs1558726 (combined P = 1.02 × 10(-5)), rs2388639 (combined P = 8.56 × 10(-9)), and rs10044254 (combined P = 9.16 × 10(-8)) independently replicated. However, these 3 SNPs were not additionally replicated in the adult asthmatic patients of the remaining trials. rs10044254 lies in the intronic region of F-box and leucine-rich repeat protein 7 (FBXL7) and is associated with decreased expression in immortalized B cells derived from CAMP participants. We have identified a novel SNP, rs10044254, associated with both decreased expression of FBXL7 and improved symptomatic response to ICSs in 2 independent pediatric cohorts. Our results suggest that there might be a specific genetic mechanism regulating symptomatic response to ICSs in children that does not carry over to adults.


What are the limits to the obligations of the nurse?
This paper enquires into the nature and the extent of the obligations of nurses. It is argued that nurses appear to be obliged to undertake supererogatory acts if they take clause one of the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) Code of Professional Conduct seriously (as, indeed, they are required to do). In the first part of the paper, the nature of nursing obligations is outlined, and then the groups and individuals to whom nurses have obligations are identified. Following a brief discussion of the moral foundation of the nurse's obligations to her/his employer, a common conflict of obligations is identified. Then a distinction is drawn between ordinary and extraordinary moral standards. Appreciation of this is necessary for an understanding of the criterion of what constitutes a supererogatory act. By the definition of supererogatory acts proposed below, it is suggested that actions such as whistleblowing satisfy that definition.


Medial tibial stress syndrome.
MTSS is a benign, though painful, condition, and a common problem in the running athlete. It is prevalent among military personnel, runners, and dancers, showing an incidence of 4% to 35%. Common names for this problem include shin splints, soleus syndrome, tibial stress syndrome, and periostitis. The exact cause of this condition is unknown. Previous theories included an inflammatory response of the periosteum or periosteal traction reaction. More recent evidence suggests a painful stress reaction of bone. The most proven risk factors are hyperpronation of the foot, female sex, and history of previous MTSS. Patient evaluation is based on meticulous history taking and physical examination. Even though the diagnosis remains clinical, imaging studies, such as plain radiographs and bone scans are usually sufficient, although MRI is useful in borderline cases to rule out more significant pathology. Conservative treatment is almost always successful and includes several options; though none has proven more superior to rest. Prevention programs do not seem to influence the rate of MTSS, though shock-absorbing insoles have reduced MTSS rates in military personnel, and ESWT has shortened the duration of symptoms. Surgery is rarely indicated but has shown some promising results in patients who have not responded to all conservative options.


Changes in physiological responses of an Antarctic fish, the emerald rock cod (Trematomus bernacchii), following exposure to polybrominated diphenyl ethers (PBDEs).
Although polybrominated diphenyl ethers (PBDEs) have the ability to undergo long-range atmospheric transport to remote ecosystems like Antarctica, a recent study found evidence for a local source within the Antarctic. PBDEs from sewage treatment outfalls of McMurdo Station and Scott Base on Ross Island have been attributed to the high concentrations measured in emerald rock cod (Trematomus bernacchii). The potential impact of PBDEs on Antarctic fish physiology is unknown and therefore, the aim of this study was to obtain a greater understanding of physiological responses of emerald rock cod for assessing changes in ecosystem quality. A PBDE mixture (ΣPBDE 8 congeners) was administered fortnightly over 42 days and physiological changes were observed throughout this period and for a further 14 days thereafter. Changes in liver composition, molecular level changes and enzyme activities of selected detoxification-mediated and antioxidant defence markers were measured. Changes in total lipid, lipid peroxide and protein carbonyl concentrations in emerald rock cod liver were consistent with increases in nucleus surface area in the PBDE-treated groups, suggesting alterations in cellular function. Changes in the activities of selected antioxidant enzymes indirectly indicated oxidative stress, possibly resulting in the changes in liver composition. Additionally, glutathione-S-transferase (GST) activity reached its peak faster than that of ethoxyresorufin-O-deethylase (EROD), suggesting that during the early response to PBDE exposures there could be a greater involvement of GST-mediated detoxification. Thus, for at least the species examined here, protein carbonyl and lipid peroxides were useful and informative biomarkers for cellular level responses following PBDE-related exposure. Furthermore, our findings suggest that emerald rock cod exposed to PBDEs develop oxidative stress - a condition with potential consequences for fish growth, health and reproduction.


Psychosocial Pretreatment Predictors of Weight Control: A Systematic Review Update.
Systematically identifying pretreatment characteristics that predict successful weight management is important to improve intervention efficacy and clinical practice. This study provides a comprehensive update of a 2005 review on pretreatment predictors of successful weight management. Results of 37 recent original studies from peer-reviewed journals were merged with the results from the 2005 review. A critical appraisal of the 66 studies included was provided, and meta-analyses were performed when feasible. Fewer previous weight loss attempts were the most consistent pretreatment predictor of successful weight management, although with a small effect size. Importantly, several variables were identified as non-significant predictors of weight loss, showing trivial effects (e.g., eating self-efficacy). Many psychosocial factors remain too little studied to allow reliable conclusions regarding their predictive value. Previous dieting attempts were identified as the soundest predictor of successful weight management. Several factors, previously considered barriers to successful weight management and now identified as non-predictors, require more investigation given the limitations identified in this review. Importantly, due to a comparably thin empirical basis for many predictors, further research is essential to move the field forward. Implications of the current state of research and necessary steps to improve intervention efficacy and clinical practice are discussed.


Aminoglycoside resistance rates, phenotypes, and mechanisms of Gram-negative bacteria from infected patients in upper Egypt.
With the re-emergence of older antibiotics as valuable choices for treatment of serious infections, we studied the aminoglycoside resistance of Gram-negative bacteria isolated from patients with ear, urinary tract, skin, and gastrointestinal tract infections at Minia university hospital in Egypt. Escherichia coli (mainly from urinary tract and gastrointestinal tract infections) was the most prevalent isolate (28.57%), followed by Pseudomonas aeruginosa (25.7%) (mainly from ear discharge and skin infections). Isolates exhibited maximal resistance against streptomycin (83.4%), and minimal resistance against amikacin (17.7%) and intermediate degrees of resistance against neomycin, kanamycin, gentamicin, and tobramycin. Resistance to older aminoglycosides was higher than newer aminoglycosides. The most common aminoglycoside resistance phenotype was that of streptomycin resistance, present as a single phenotype or in combination, followed by kanamycin-neomycin as determined by interpretative reading. The resistant Pseudomonas aeruginosa strains were capable of producing aminoglycoside-modifying enzymes and using efflux as mechanisms of resistance. Using checkerboard titration method, the most frequently-observed outcome in combinations of aminoglycosides with β-lactams or quinolones was synergism. The most effective combination was amikacin with ciprofloxacin (100% Synergism), whereas the least effective combination was gentamicin with amoxicillin (53.3% Synergistic, 26.7% additive, and 20% indifferent FIC indices). Whereas the studied combinations were additive and indifferent against few of the tested strains, antagonism was never observed. The high resistance rates to aminoglycosides exhibited by Gram-negative bacteria in this study could be attributed to the selective pressure of aminoglycoside usage which could be controlled by successful implementation of infection control measures.


Demonstration of opsonic activity and in vivo protection against group B streptococci type III by Streptococcus pneumoniae type 14 antisera.
The present studies demonstrate that antisera directed against Streptococcus pneumoniae type 14 is opsonic for group B streptococci type III in a neutrophile-mediated bactericidal assay. Specificity was demonstrated by the observations that group B streptococci type III and S. pneumoniae type 14 adsorbed the opsonic activity of anti-S. pneumoniae type 14 antisera. Group B streptococci strain 090R (devoid of type antigens) and S. pneumoniae type 3, did not remove the opsonic activity of anti-S. pneumoniae type 14 serum. In vivo studies using a suckling rat model of neonatal group B streptococcal type III sepsis demonstrated that antisera directed against S. pneumoniae type 14 was highly protective.


Energy-dependent chromatin remodelers: complex complexes and their components.
Chromatin structure is dynamically regulated such that it can be modified by a number of factors in response to a variety of signals. One class of factors that can mediate changes in chromatin structure is the ATP-dependent nucleosome remodeling complexes. Genetic and biochemical evidence supports the idea that a family of related multisubunit complexes hydrolyzes ATP in order to facilitate the rearrangement of chromatin structure. These complexes are conserved from yeast to mammals and apparently have diverse functions in modifying chromatin structure; ATP-dependent chromatin remodelers have been implicated in nucleosome deposition, nucleosome assembly, and disruption of nucleosome structure to facilitate transcriptional activation. In addition, individual components of these complexes have been linked to control of cell growth, cell cycle regulation, development, and differentiation, and they may also be targets for viral regulatory proteins. The diversity of subunit functions likely relates to effects on chromatin structure, suggesting that the regulation of chromatin structure by ATP-dependent remodelers is important in many different aspects of cellular metabolism.


The potential role of taxes and subsidies on food in the prevention of obesity in Europe.
Obesity implies costs not only for the individual but also for society. The authors explore the opinions of stakeholders on the potential of taxes or subsidies, as measures for tackling obesity in Europe. Structured interviews were conducted using Multicriteria Mapping, a computer-based, decision-support tool, with 189 interviewees drawn from 21 different stakeholder categories across nine members of the EU interviews, to appraise 20 predefined policy options aimed at reducing obesity, including 'taxing obesity-promoting foods' and 'subsidising healthy foods.' A four-step approach involved selecting options, defining criteria, scoring options quantitatively and weighting criteria to provide overall rankings of options. Interviews were recorded and transcribed to yield qualitative data. Compared with other policy options appraised, taxation and subsidies were not favourably received, mainly because they were considered difficult to implement. Overall, trade unions rated both options more favourably than all other stakeholder groups. As anticipated, both options received their lowest scores from representatives of the farming, food processing and advertising industries. Nutritional/obesity advisory experts and public sector caterers gave the most positive ratings to subsidies overall. Along with public health professionals, large commercial retailers were most in favour of taxation. Taxation and subsidies were poorly appraised compared with other policy measures, with stakeholders expressing reservations mainly focussed on the practicalities and cost of introducing such measures. Although applying taxes/subsidies could be useful to combat obesity, the study suggests that most stakeholders still need to be convinced of their viability and acceptability when compared with other measures.


GSK3 Inhibition Drives Maturation of NK Cells and Enhances Their Antitumor Activity.
Maturation of human natural killer (NK) cells as defined by accumulation of cell-surface expression of CD57 is associated with increased cytotoxic character and TNF and IFNγ production upon target-cell recognition. Notably, multiple studies point to a unique role for CD57+ NK cells in cancer immunosurveillance, yet there is scant information about how they mature. In this study, we show that pharmacologic inhibition of GSK3 kinase in peripheral blood NK cells expanded ex vivo with IL15 greatly enhances CD57 upregulation and late-stage maturation. GSK3 inhibition elevated the expression of several transcription factors associated with late-stage NK-cell maturation including T-BET, ZEB2, and BLIMP-1 without affecting viability or proliferation. When exposed to human cancer cells, NK cell expanded ex vivo in the presence of a GSK3 inhibitor exhibited significantly higher production of TNF and IFNγ, elevated natural cytotoxicity, and increased antibody-dependent cellular cytotoxicity. In an established mouse xenograft model of ovarian cancer, adoptive transfer of NK cells conditioned in the same way also displayed more robust and durable tumor control. Our findings show how GSK3 kinase inhibition can greatly enhance the mature character of NK cells most desired for effective cancer immunotherapy. Cancer Res; 77(20); 5664-75. ©2017 AACR.


Global environmental costs of China's thirst for milk.
China has an ever-increasing thirst for milk, with a predicted 3.2-fold increase in demand by 2050 compared to the production level in 2010. What are the environmental implications of meeting this demand, and what is the preferred pathway? We addressed these questions by using a nexus approach, to examine the interdependencies of increasing milk consumption in China by 2050 and its global impacts, under different scenarios of domestic milk production and importation. Meeting China's milk demand in a business as usual scenario will increase global dairy-related (China and the leading milk exporting regions) greenhouse gas (GHG) emissions by 35% (from 565 to 764 Tg CO2eq ) and land use for dairy feed production by 32% (from 84 to 111 million ha) compared to 2010, while reactive nitrogen losses from the dairy sector will increase by 48% (from 3.6 to 5.4 Tg nitrogen). Producing all additional milk in China with current technology will greatly increase animal feed import; from 1.9 to 8.5 Tg for concentrates and from 1.0 to 6.2 Tg for forage (alfalfa). In addition, it will increase domestic dairy related GHG emissions by 2.2 times compared to 2010 levels. Importing the extra milk will transfer the environmental burden from China to milk exporting countries; current dairy exporting countries may be unable to produce all additional milk due to physical limitations or environmental preferences/legislation. For example, the farmland area for cattle-feed production in New Zealand would have to increase by more than 57% (1.3 million ha) and that in Europe by more than 39% (15 million ha), while GHG emissions and nitrogen losses would increase roughly proportionally with the increase of farmland in both regions. We propose that a more sustainable dairy future will rely on high milk demanding regions (such as China) improving their domestic milk and feed production efficiencies up to the level of leading milk producing countries. This will decrease the global dairy related GHG emissions and land use by 12% (90 Tg CO2eq reduction) and 30% (34 million ha land reduction) compared to the business as usual scenario, respectively. However, this still represents an increase in total GHG emissions of 19% whereas land use will decrease by 8% when compared with 2010 levels, respectively.


The effectiveness of continuing medical education for specialist recertification.
Evolving professional, social and political pressures highlight the importance of lifelong learning for clinicians. Continuing medical education (CME) facilitates lifelong learning and is a fundamental factor in the maintenance of certification. The type of CME differs between surgical and non-surgical specialties. CME methods of teaching include lectures, workshops, conferences and simulation training. Interventions involving several modalities, instructional techniques and multiple exposures are more effective. The beneficial effects of CME can be maintained in the long term and can improve clinical outcome. However, quantitative evidence on validity, reliability, efficacy and cost-effectiveness of various methods is lacking. This is especially evident in urology. The effectiveness of CME interventions on maintenance of certification is also unknown. Currently, many specialists fulfil mandatory CME credit requirements opportunistically, therefore erroneously equating number of hours accumulated with competence. New CME interventions must emphasize actual performance and should correlate with clinical outcomes. Improved CME practice must in turn lead to continuing critical reflection, practice modification and implementation with a focus towards excellent patient care.


Genome analysis of a novel, highly divergent picornavirus from common kestrel (Falco tinnunculus): the first non-enteroviral picornavirus with type-I-like IRES.
Although the number of identified avian-borne picornaviruses (family Picornaviridae) is continuously increasing there remains several species-rich avian host groups, such as the order Falconiformes (with 290 bird species) from which picornaviruses have not been identified. This study reports the first complete genome of a novel, highly divergent picornavirus, named as Falcovirus A1 (KP230449), from the carnivorous bird, the common kestrel (Falco tinnunculus, order Falconiformes). Falcovirus A1 has the longest 3D(RdRp) genome region and distant phylogenetic relationship to the Hepatitis A virus 1 (Hepatovirus) and Avian encephalomyelitis virus 1 (Tremovirus). It has a type-I (enterovirus-like) IRES in the 5'UTR - identified for the first time among avian-borne picornaviruses suggesting that type-I IRES is not restricted only to enteroviruses and providing further evidence of mosaicism of this region among different picornavirus genera.


New Therapeutic Horizons for Graves' Hyperthyroidism.
Graves' hyperthyroidism is characterized by the presence of autoantibodies that stimulate the thyroid-stimulating hormone receptor (TSHR), resulting in uncontrolled secretion of excessive thyroid hormone. Conventional treatments, including antithyroid medication, radioiodine, or surgery have remained largely unchanged for the past 70 years and either lack efficacy for many patients, or result in lifelong thyroid hormone replacement therapy, in the case of the latter 2 options. The demand for new therapeutic options, combined with greater insight into basic immunobiology, has led to the emergence of novel approaches to treat Graves' hyperthyroidism. The current therapies under investigation include biologics, small molecules, and peptide immunomodulation. There is a growing focus on TSHR-specific treatment modalities, which carry the advantage of eliciting a specific, targeted approach, with the aim of avoiding disruption of the functioning immune system. These therapies present a new opportunity to supersede the inadequate treatments currently available for some Graves' patients, offering hope of successful restoration of euthyroidism without the need for ongoing therapy. Several of these therapeutic options have the potential to translate into clinical practice in the near future. This review provides a comprehensive summary of the recent advances and various stages of development of the novel therapeutic approaches to treat Graves' hyperthyroidism.


Rickettsia rickettsii Whole-Cell Antigens Offer Protection against Rocky Mountain Spotted Fever in the Canine Host.
Rocky Mountain spotted fever (RMSF) is a potentially fatal tick-borne disease in people and dogs. RMSF is reported in the United States and several countries in North, Central, and South America. The causative agent of this disease, Rickettsia rickettsii, is transmitted by several species of ticks, including Dermacentor andersoni, Rhipicephalus sanguineus, and Amblyomma americanum RMSF clinical signs generally include fever, headache, nausea, vomiting, muscle pain, lack of appetite, and rash. If untreated, it can quickly progress into a life-threatening illness in people and dogs, with high fatality rates ranging from 30 to 80%. While RMSF has been known for over a century, recent epidemiological data suggest that the numbers of documented cases and the fatality rates remain high in people, particularly during the last two decades in parts of North America. Currently, there are no vaccines available to prevent RMSF in either dogs or people. In this study, we investigated the efficacies of two experimental vaccines, a subunit vaccine containing two recombinant outer membrane proteins as recombinant antigens (RCA) and a whole-cell inactivated antigen vaccine (WCA), in conferring protection against virulent R. rickettsii infection challenge in a newly established canine model for RMSF. Dogs vaccinated with WCA were protected from RMSF, whereas those receiving RCA developed disease similar to that of nonvaccinated R. rickettsii-infected dogs. WCA also reduced the pathogen loads to nearly undetected levels in the blood, lungs, liver, spleen, and brain and induced bacterial antigen-specific immune responses. This study provides the first evidence of the protective ability of WCA against RMSF in dogs.