Neurocognitive dysfunction in hematopoietic cell transplant recipients: expert review from the late effects and Quality of Life Working Committee of the CIBMTR and complications and Quality of Life Working Party of the EBMT.
Hematopoietic cell transplantation (HCT) is a potentially curative treatment for children and adults with malignant and non-malignant diseases. Despite increasing survival rates, long-term morbidity following HCT is substantial. Neurocognitive dysfunction is a serious cause of morbidity, yet little is known about neurocognitive dysfunction following HCT. To address this gap, collaborative efforts of the Center for International Blood and Marrow Transplant Research and the European Society for Blood and Marrow Transplantation undertook an expert review of neurocognitive dysfunction following HCT. In this review, we define what constitutes neurocognitive dysfunction, characterize its risk factors and sequelae, describe tools and methods to assess neurocognitive function in HCT recipients, and discuss possible interventions for HCT patients with this condition. This review aims to help clinicians understand the scope of this health-related problem, highlight its impact on well-being of survivors, and to help determine factors that may improve identification of patients at risk for declines in cognitive functioning after HCT. In particular, we review strategies for preventing and treating neurocognitive dysfunction in HCT patients. Lastly, we highlight the need for well-designed studies to develop and test interventions aimed at preventing and improving neurocognitive dysfunction and its sequelae following HCT.


Surface-plasmon-resonance sensor based on three-hole microstructured optical fiber.
We propose a novel surface-plasmon-resonance sensor design based on coating the holes of a three-hole microstructured optical fiber with a low-index dielectric layer on top of which a gold layer is deposited. The use of all three fiber holes and their relatively large size should facilitate the fabrication of the inclusions and the infiltration of the analyte. Our numerical results indicate that the optical loss of the Gaussian guided mode can be made very small by tuning the thickness of the dielectric layer and that the refractive-index resolution for aqueous analytes is 1x 10(-4).


Identification and characterization of genes required for biosynthesis and transport of the siderophore vibrioferrin in Vibrio parahaemolyticus.
In response to low iron availability, Vibrio parahaemolyticus synthesizes and secretes a polyhydroxycarboxylate-type siderophore vibrioferrin which is composed of 1 mol each of 2-ketoglutaric acid, L-alanine, ethanolamine, and citric acid. We have previously reported the cloning and characterization of the pvuA gene, which encodes the 78-kDa outer membrane receptor protein for ferric vibrioferrin. In this study, nine genes involved in the biosynthesis and transport of vibrioferrin have been identified in the genomic regions surrounding the pvuA gene. The genes were sequenced, and gene disruptants were constructed by insertion mutation for phenotype analysis. Five of the genes, named pvsABCDE, constitute an operon that is expressed under iron-limiting conditions. Homology searches of their predicted protein products suggested that the four genes pvsABDE are implicated in the biosynthesis of the siderophore. Another gene in the same operon, pvsC, encodes a putative exporter that is homologous to members of the major facilitator superfamily of multidrug efflux pumps. The remaining four genes, named pvuBCDE, encode proteins strongly homologous to Escherichia coli FecBCDE, respectively, which are components of the ATP-binding cassette transporter system for ferric dicitrate. Reverse transcriptase PCR analysis revealed that these transport genes are transcribed as a single mRNA with the upstream genes, psuA and pvuA. Phenotypic comparison between the wild-type strain and its targeted gene disruptants supported the biological functions for the respective operons that were expected on the basis of the homology search.


A simple score for estimating the long-term risk of fracture in patients with multiple sclerosis.
To derive a simple score for estimating the long-term risk of osteoporotic and hip fracture in individual patients with MS. Using the UK General Practice Research Database linked to the National Hospital Registry (1997-2008), we identified patients with incident MS (n = 5,494). They were matched 1:6 by year of birth, sex, and practice with patients without MS (control subjects). Cox proportional hazards models were used to calculate the long-term risk of osteoporotic and hip fracture. We fitted the regression model with general and specific risk factors, and the final Cox model was converted into integer risk scores. In comparison with the FRAX calculator, our risk score contains several new risk factors that have been linked with fracture, which include MS, use of antidepressants, use of anticonvulsants, history of falling, and history of fatigue. We estimated the 5- and 10-year risks of osteoporotic and hip fracture in relation to the risk score. The C-statistic was moderate (0.67) for the prediction of osteoporotic fracture and excellent (0.89) for the prediction of hip fracture. This is the first clinical risk score for fracture risk estimation involving MS as a risk factor.


Reduced repair capacity of a DNA clustered damage site comprised of 8-oxo-7,8-dihydro-2'-deoxyguanosine and 2-deoxyribonolactone results in an increased mutagenic potential of these lesions.
A signature of ionizing radiation is the induction of DNA clustered damaged sites. Non-double strand break (DSB) clustered damage has been shown to compromise the base excision repair pathway, extending the lifetimes of the lesions within the cluster, compared to isolated lesions. This increases the likelihood the lesions persist to replication and thus increasing the mutagenic potential of the lesions within the cluster. Lesions formed by ionizing radiation include 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) and 2-deoxyribonolactone (dL). dL poses an additional challenge to the cell as it is not repaired by the short-patch base excision repair pathway. Here we show recalcitrant dL repair is reflected in mutations observed when DNA containing it and a proximal 8-oxodGuo is replicated in Escherichia coli. 8-oxodGuo in close proximity to dL on the opposing DNA strand results in an enhanced frequency of mutation of the lesions within the cluster and a 20 base sequence flanking the clustered damage site in an E. coli based plasmid assay. In vitro repair of a dL lesion is reduced when compared to the repair of an abasic (AP) site and a tetrahydrofuran (THF), and this is due mainly to a reduction in the activity of polymerase β, leading to retarded FEN1 and ligase 1 activities. This study has given insights in to the biological effects of clusters containing dL.


Pediatricians' and family physicians' weight-related care of children in the U.S.
Few national data exist to assess primary care physicians' (PCPs') clinical practices with regard to childhood obesity. To survey pediatricians and family practice physicians regarding their assessment, counseling, and management of diet, physical activity, and weight status among pediatric patients in the primary care setting. A nationally representative cross-sectional survey of pediatricians and family practice physicians sampled from the American Medical Association (AMA) Masterfile was conducted in 2008 and analyzed in 2010. Outcomes included physicians' self-reported practice behaviors regarding assessments of pediatric patients' weight status, counseling of diet and physical activity, and referrals and follow-ups. Response rate excluding physicians listed as "no-contact" by the AMA was 73.7% among pediatricians and 66.9% among family physicians. Less than 50% of all PCPs assessed BMI percentiles regularly in children. Eighteen percent of all PCPs reported referring children for further evaluation or management. Fifty-eight percent of all PCPs reported never, rarely, or only sometimes tracking patients over time concerning weight or weight-related behaviors. Pediatricians were more likely than family physicians to assess weight status and provide behavioral counseling (p's<0.001). Active PCP participation in assessing or managing childhood obesity in the primary care setting appears low relative to the frequency of the problem in the U.S. Interventions to reduce the barriers to physician engagement in the assessment and management of healthy lifestyles are needed to prevent and control childhood obesity.


Leptospira and Leptospirosis.
Leptospira spp. are morphologically and phylogenetically unique bacteria that are ubiquitous in the environment. Pathogenic species are the agents of leptospirosis, an emerging zoonotic disease transmitted to both human and animals from the environment contaminated with the urine of reservoir animals. The taxonomy of Leptospira has recently undergone extensive revisions with the use of whole-genome sequences, thus expanding considerably the number of species.


Bone marrow stromal cells as replacement cells for Parkinson's disease: generation of an anatomical but not functional neuronal phenotype.
The focus of cell replacement therapies (CRTs) for Parkinson's disease has been on delivering dopamine-producing cells to the striatum. Fetal grafts have proven the feasibility of this approach, but an appropriate source of replacement cells has restricted the clinical translation. Bone marrow stromal cells (BMSCs) have been heralded as an ideal source of dopaminergic (DAergic) replacement cells, as they are viewed as ethically acceptable, easily procured, and readily expanded. It is known that they confer functional benefits, particularly in stroke models, through the release of neurotrophic factors, but their transdifferentiation into neurons is still under contention. We sought to evaluate the neuronal phenotype and functional capacity of adult rat BMSCs after exposure to a novel multistep in vitro differentiation protocol compared with cells exposed to other reported neuronal differentiation conditions. We employed a systematic, comprehensive method of assessment to determine the neuronal differentiation capacity of BMSCs. Our fluorescence-activated cell sorting, immunofluorescent and semiquantitative polymerase chain reaction results confirmed that undifferentiated BMSCs isolated based on their adherence to plastic are of mesenchymal origin and express a range of lineage markers. After exposure to preinduction and neuronal induction steps, BMSCs down-regulate markers of other lineages but fail, as assessed by patch clamp, to differentiate into functional neurons. Thus, for BMSCs to be considered a source of DAergic neuronal replacement cells, their ability to transdifferentiate terminally along a neuronal lineage first must be clarified before attempting to direct more complex specification process required for them to be used in Parkinson's-disease-focused CRTs.


Assessing the risk of exogenously consumed pharmaceuticals in land-applied human urine.
Once separated, the use of urine as fertilizer is a particular attractive proposition and can significantly mitigate the release of nutrients and pharmaceutically active compounds (PhACs) to the environment. In the current study, a simple methodological framework is proposed for assessing risks that are posed by the land application of urine, which contains PhACs, in terms of 6 selected environmental and human-health endpoints. In total, 25 commonly used PhACs were conservatively assessed using the proposed methodology and results indicated that 14 of them may pose a risk with respect to either eco-toxicological or human-health endpoints. The receiving terrestrial environment was identified as the most susceptible of the eco-toxicological endpoints and hazard to human-health was most significant through food-chain transfer. The results highlight the need to consider the potential impacts associated with pharmaceuticals and the need to pre-treat urine to address the presence of problematic PhACs before it is applied on land.


Pressure-controlled formation of crystalline, Janus, and core-shell supraparticles.
Binary mixtures of nanoparticles self-assemble in the confinement of evaporating oil droplets and form regular supraparticles. We demonstrate that moderate pressure differences on the order of 100 kPa change the particles' self-assembly behavior. Crystalline superlattices, Janus particles, and core-shell particle arrangements form in the same dispersions when changing the working pressure or the surfactant that sets the Laplace pressure inside the droplets. Molecular dynamics simulations confirm that pressure-dependent interparticle potentials affect the self-assembly route of the confined particles. Optical spectrometry, small-angle X-ray scattering and electron microscopy are used to compare experiments and simulations and confirm that the onset of self-assembly depends on particle size and pressure. The overall formation mechanism reminds of the demixing of binary alloys with different phase diagrams.


Reconsidering the relationship between fast-food outlets, area-level deprivation, diet quality and body mass index: an exploratory structural equation modelling approach.
Internationally, the prevalence of adults with obesity is a major public health concern. Few studies investigate the explanatory pathways between fast-food outlets and body mass index (BMI). We use structural equation modelling to explore an alternative hypothesis to existing research using area-level deprivation as the predictor of BMI and fast-food outlets and diet quality as mediators. Adults (n=7544) from wave II of the Yorkshire Health Study provided self-reported diet, height and weight (used to calculate BMI). Diet quality was based on sugary drinks, wholemeal (wholegrain) bread and portions of fruit and vegetables. Fast-food outlets were mapped using the Ordnance Survey Points of Interest within 2 km radial buffers around home postcode which were summed to indicate availability. Age (years), gender (female/male) and long-standing health conditions (yes/no) were included as covariates. There was little evidence linking fast-food outlets to diet or BMI. An independent association between fast-food outlet availability and BMI operated counterintuitively and was small in effect. There was also little evidence of mediation between fast-food outlet availability and BMI. However, there was more evidence that area-level deprivation was associated with increased BMI, both as an independent effect and through poorer diet quality. This exploratory study offers a first step for considering complexity and pathways linking fast-food outlets, area-level deprivation, diet quality and BMI. Research should respond to and build on the hypothesised pathways and our simple framework presented within our study.


Movement Enhances the Nonlinearity of Hippocampal Theta.
The nonlinear, metastable dynamics of the brain are essential for large-scale integration of smaller components and for the rapid organization of neurons in support of behavior. Therefore, understanding the nonlinearity of the brain is paramount for understanding the relationship between brain dynamics and behavior. Explicit quantitative descriptions of the properties and consequences of nonlinear neural networks, however, are rare. Because the local field potential (LFP) reflects the total activity across a population of neurons, nonlinearites of the nervous system should be quantifiable by examining oscillatory structure. We used high-order spectral analysis of LFP recorded from the dorsal and intermediate regions of the rat hippocampus to show that the nonlinear character of the hippocampal theta rhythm is directly related to movement speed of the animal. In the time domain, nonlinearity is expressed as the development of skewness and asymmetry in the theta shape. In the spectral domain, nonlinear dynamics manifest as the development of a chain of harmonics statistically phase coupled to the theta oscillation. This evolution was modulated across hippocampal regions, being stronger in the dorsal CA1 relative to more intermediate areas. The intensity and timing of the spiking activity of pyramidal cells and interneurons was strongly correlated to theta nonlinearity. Because theta is known to propagate from dorsal to ventral regions of the hippocampus, these data suggest that the nonlinear character of theta decreases as it travels and supports a hypothesis that activity dissipates along the longitudinal axis of the hippocampus. We describe the first explicit quantification regarding how behavior enhances the nonlinearity of the nervous system. Our findings demonstrate uniquely how theta changes with increasing speed due to the altered underlying neuronal dynamics and open new directions of research on the relationship between single-neuron activity and propagation of theta through the hippocampus. This work is significant because it will encourage others to consider the nonlinear nature of the nervous system and higher-order spectral analyses when examining oscillatory interactions.


Recurrent De Novo Dominant Mutations in SLC25A4 Cause Severe Early-Onset Mitochondrial Disease and Loss of Mitochondrial DNA Copy Number.
Mutations in SLC25A4 encoding the mitochondrial ADP/ATP carrier AAC1 are well-recognized causes of mitochondrial disease. Several heterozygous SLC25A4 mutations cause adult-onset autosomal-dominant progressive external ophthalmoplegia associated with multiple mitochondrial DNA deletions, whereas recessive SLC25A4 mutations cause childhood-onset mitochondrial myopathy and cardiomyopathy. Here, we describe the identification by whole-exome sequencing of seven probands harboring dominant, de novo SLC25A4 mutations. All affected individuals presented at birth, were ventilator dependent and, where tested, revealed severe combined mitochondrial respiratory chain deficiencies associated with a marked loss of mitochondrial DNA copy number in skeletal muscle. Strikingly, an identical c.239G>A (p.Arg80His) mutation was present in four of the seven subjects, and the other three case subjects harbored the same c.703C>G (p.Arg235Gly) mutation. Analysis of skeletal muscle revealed a marked decrease of AAC1 protein levels and loss of respiratory chain complexes containing mitochondrial DNA-encoded subunits. We show that both recombinant AAC1 mutant proteins are severely impaired in ADP/ATP transport, affecting most likely the substrate binding and mechanics of the carrier, respectively. This highly reduced capacity for transport probably affects mitochondrial DNA maintenance and in turn respiration, causing a severe energy crisis. The confirmation of the pathogenicity of these de novo SLC25A4 mutations highlights a third distinct clinical phenotype associated with mutation of this gene and demonstrates that early-onset mitochondrial disease can be caused by recurrent de novo mutations, which has significant implications for the application and analysis of whole-exome sequencing data in mitochondrial disease.


Quantitative verification of the keyhole concept: a comparison of area of exposure in the parasellar region via supraorbital keyhole, frontotemporal pterional, and supraorbital approaches.
This study was designed to determine if the "keyhole concept," proposed by Perneczky's group, can be verified quantitatively. Fourteen (3 bilateral and 8 unilateral) sides of embalmed latex-injected cadaveric heads were dissected via 3 sequential craniotomy approaches: supraorbital keyhole, frontotemporal pterional, and supraorbital. Three-dimensional cartesian coordinates were recorded using a stereotactic localizer. The orthocenter of the ipsilateral anterior clinoid process, the posterior clinoid process, and the contralateral anterior clinoid process are expressed as a center point (the apex). Seven vectors project from the apex to their corresponding target points in a radiating manner on the parasellar skull base. Each 2 neighboring vectors border what could be considered a triangle, and the total area of the 7 triangles sharing the same apex was geometrically expressed as the area of exposure in the parasellar region. Values are expressed as the mean ± SD (mm(2)). The total area of exposure was as follows: supraorbital keyhole 1733.1 ± 336.0, pterional 1699.3 ± 361.9, and supraorbital 1691.4 ± 342.4. The area of exposure on the contralateral side was as follows: supraorbital keyhole 602.2 ± 194.7, pterional 595.2 ± 228.0, and supraorbital 553.3 ± 227.2. The supraorbital keyhole skull flap was 2.0 cm(2), and the skull flap size ratio was 1:5:6.5 (supraorbital keyhole/pterional/supraorbital). The area of exposure of the parasellar region through the smaller supraorbital keyhole approach is as adequate as the larger pterional and supraorbital approaches. The keyhole concept can be verified quantitatively as follows: 1) a wide area of exposure on the skull base can be obtained through a small keyhole skull opening, and 2) the side opposite the opening can also be visualized.


Meeting need vs. sharing the market: a systematic review of methods to measure the use of private sector family planning and childbirth services in sub-Saharan Africa.
Ensuring universal access to maternal and reproductive health services is critical to the success of global efforts to reduce poverty and inequality. Engaging private providers has been proposed as a strategy for increasing access to healthcare in low- and middle-income countries; however, little consensus exists on how to estimate the extent of private sector use. Using research from sub-Saharan Africa, this study systematically compares and critiques quantitative measures of private sector family planning and childbirth service use and synthesizes evidence on the role of the private sector in the region. We conducted a systematic review of the Medline, Global Health, and Popline databases. All studies that estimated use of private sector of family planning or childbirth services in one or more sub-Saharan African countries were included in this review. For each study, we extracted data on the key study outcomes and information on the methods used to estimate private sector use. Fifty-three papers met our inclusion criteria; 31 provided outcomes on family planning, and 26 provided childbirth service outcomes. We found substantial methodological variation between studies; for instance, while some reported on service use from any private sector source, others distinguished private sector providers either by their profit orientation or position within or outside the formal medical sector. Additionally, studies measured the use of private sector services differently, with some estimating the proportion of need met by the private sector and others examining the sector's share among the market of service users. Overall, the estimates suggest that the private sector makes up a considerable portion (> 20%) of the market for family planning and childbirth care, but its role in meeting women's need for these services is fairly low (< 10%). Many studies have examined the extent of private sector family planning and childbirth service provision; however, inconsistent methodologies make it difficult to compare results across studies and contexts. Policymakers should consider the implications of both private market share and coverage estimates, and be cautious in interpreting data on the scale of private sector health service provision without a clear understanding of the methodology.


Prioritisation of infectious diseases in public health: feedback on the prioritisation methodology, 15 July 2008 to 15 January 2009.
In 2004, the German public health institute, the Robert Koch Institute (RKI), prioritised pathogens by public health criteria and presented the methodology and findings. In order to further improve the methodology, the RKI invited experts to give feedback on this via a structured web-based questionnaire. The survey was completed by 72 participants during 15 July 2008 to 15 January 2009. Prioritisation of pathogens was considered as useful for public health purposes by 68 participants and for both surveillance and epidemiological research by 64 participants. Additional pathogens were suggested, including some that are resistant to antimicrobials. The criteria incidence, severity, outbreak potential, emerging potential and preventability were each considered as useful or very useful for the prioritisation (by more than 65 participants for each criterion). Weighting of the criteria was judged as relevant or very relevant by 67 of participants, but needs more explanation. It was also suggested that the group carrying out the prioritisation be composed of a median of 15 experts (range: 5–1,000). The feedback obtained in the survey has been taken into account in the modification of the methodology for the next round of prioritisation, which started in December 2010.


A randomized clinical trial of energy conservation for patients with cancer-related fatigue.
The efficacy of energy conservation and activity management (ECAM) for fatigue reduction and maintenance of functional performance has never been evaluated in adults with cancer who are undergoing treatment. A randomized clinical trial compared an ECAM intervention with a control intervention focused on nutrition. Individuals initiating chemotherapy, radiotherapy, or concurrent therapy for cancer were randomized to receive either the semistructured ECAM intervention (n = 200) or the control intervention (n = 196). Participants in each group participated in 3 telephone sessions with an oncology nurse during the first 5 weeks of treatment. Data on fatigue and limitation of functioning were obtained before cancer treatment and at two follow-up points that coincided with times of high fatigue for each type of treatment. The outcomes of interest included perception of fatigue and functional performance. A repeated-measures analysis of covariance using the type of cancer treatment as a covariate revealed a significant study group-by-time interaction indicating that the ECAM group experienced a greater decrease in fatigue over time compared with the control group (F(2,544) = 4.5; P = 0.01). The intervention was not associated with changes in overall functional performance. Individuals who received the ECAM intervention derived a modest but significant benefit from it. To achieve a more robust clinical benefit from the intervention, it may be necessary to manage other key symptoms in addition to fatigue. Research is needed to examine symptom clusters or combinations associated with negative outcomes as well as combination strategies for symptom management.


Treatment of visuospatial neglect with biparietal tDCS and cognitive training: a single-case study.
Symptoms of visuospatial neglect occur frequently after unilateral brain damage. Neglect hampers rehabilitation progress and is associated with reduced quality of life. However, existing treatment methods show limited efficacy. Transcranial direct current stimulation (tDCS) is a neuromodulatory technique, which can be used to increase or decrease brain excitability. Its combination with conventional neglect therapy may enhance treatment efficacy. A 72-year-old male with a subacute ischemic stroke of the right posterior cerebral artery suffering from visuospatial neglect, hemianopia, and hemiparesis was treated with biparietal tDCS and cognitive neglect therapy in a double-blind, sham-controlled single-case study. Four weeks of daily treatment sessions (5 days per week, 30 min) were started 26 days post-stroke. During week 1 and 4 the patient received conventional neglect therapy, during week 2, conventional neglect therapy was combined once with sham and once with real biparietal tDCS. Week 3 consisted of daily sessions of real biparietal tDCS (1 mA, 20 min) combined with neglect therapy. Outcome measures were assessed before, immediately after, as well as 1 week and 3 months after the end of treatment. They included subtests of the Test for Attentional Performance (TAP): covert attention (main outcome), alertness, visual field; the Neglect-Test (NET): line bisection, cancelation, copying; and activities of daily living (ADL). After real stimulation, covert attention allocation toward left-sided invalid stimuli was significantly improved, and line bisection and copying improved qualitatively as compared to sham stimulation. ADL were only improved at the 3-month follow-up. This single-case study demonstrates for the first time that combined application of tDCS and cognitive training may enhance training-induced improvements in measures of visuospatial neglect and is applicable in a clinical context.


"The complaining women": health professionals' perceptions on patients with fibromyalgia in Spain.
The aim of this study is twofold: (1) to explore health service providers' perceptions regarding fibromyalgia patients in Spain and (2) to analyze possible consequences of these perceptions in terms of how health service providers construct the disease and treat their patients. Qualitative study. Twelve health service providers (eight men, four women) involved in the care of fibromyalgia patients. Providers were from different disciplines and included general practitioners, rheumatologists, occupational doctors, psychologists, psychiatrists, physiotherapists and behavioral specialists from Spain. We performed individual semistructured interviews, which were recorded and transcribed to conduct a qualitative content analysis supported by Atlas.ti-7. We identified three categories from the interviews: (1) the fibromyalgia patient prototype: the complaining woman, (2) fibromyalgia is considered a women's health issue, but male patients are a privileged minority, and (3) health professionals' attitudes toward fibromyalgia patients: are they really suffering or pretending? The uncertainty surrounding fibromyalgia together with the fact that those affected are primarily women, seem to influence professional practice in terms of lack of recognition of Fibromyalgia as a severe disease. Increased training of all health professionals is essential to improving the support and attention given to patients suffering from fibromyalgia. Implications for rehabilitation Fibromyalgia • In order to improve fibromyalgia patients´ attention, health providers should learn how to assist patients without prejudices. • Training programs for health providers should include sensitization about the severity of fibromyalgia. • Health providers should be aware of the existence of stereotypes about women suffering from fibromyalgia. • Fibromyalgia protocols should give skills to health providers to avoid offering a gender-biased attention to patients.


TU-E-217BCD-04: Spectral Breast CT: Effect of Adaptive Filtration on CT Numbers, CT Noise, and CNR.
Photon counting spectral breast CT is feasible in part due to using an adaptive filter. An adaptive filter provides flat x-ray intensity profile and constant x-ray energy spectrum across detector surface, decreases required detector count rate, and eliminates beam hardening artifacts. However, the altered x-ray exposure profiles at the breast and detector surface may influence the distribution of CT noise, CT numbers, and contrast to noise ratio (CNR) across the CT images. The purpose of this work was to investigate these effects. Images of a CT phantom with and without adaptive filter were simulated at 60kVp, 90kVp, and 120kVp tube voltages and 660 mR total skin exposure. The CT phantom with water content had 14cm diameter, contrast elements representing adipose tissue and 2.5mg/cc iodine contrast located at 1cm, 3.5cm, and 6cm from center of the phantom. The CT numbers, CT noise, and CNR were measured at multiple locations for several filter/exposure combinations: (1)without adaptive filter for 660mR skin exposure; (2)with adaptive filter for 660mR skin exposure along central axis (mean skin exposure across the breast was <660mR); and (3)with adaptive filter for scaled exposure (mean skin exposure was 660mR). Beam hardening (cupping) artifacts had 47HU magnitude without adaptive filter but were eliminated with adaptive filter. CNR of contrast elements was comparable for (1) and (2) over central parts but was higher by 20-30% for (1) near the edge of the phantom. CNR was higher by 20-30% in (3) as compared to (2) over central parts and comparable near the edges. The adaptive filter provided: uniform distribution of CT noise, CNR, and CT numbers across CT images; comparable or better CNR with no dose penalty to the breast; and eliminated beam hardening artifacts.


Environmental Subconcussive Injury, Axonal Injury, and Chronic Traumatic Encephalopathy.
Brain injury occurs in two phases: the initial injury itself and a secondary cascade of precise immune-based neurochemical events. The secondary phase is typically functional in nature and characterized by delayed axonal injury with more axonal disconnections occurring than in the initial phase. Axonal injury occurs across the spectrum of disease severity, with subconcussive injury, especially when repetitive, now considered capable of producing significant neurological damage consistent with axonal injury seen in clinically evident concussion, despite no observable symptoms. This review is the first to introduce the concept of environmental subconcussive injury (ESCI) and sets out how secondary brain damage from ESCI once past the juncture of microglial activation appears to follow the same neuron-damaging pathway as secondary brain damage from conventional brain injury. The immune response associated with ESCI is strikingly similar to that mounted after conventional concussion. Specifically, microglial activation is followed closely by glutamate and calcium flux, excitotoxicity, reactive oxygen species and reactive nitrogen species (RNS) generation, lipid peroxidation, and mitochondrial dysfunction and energy crisis. ESCI damage also occurs in two phases, with the primary damage coming from microbiome injury (due to microbiome-altering events) and secondary damage (axonal injury) from progressive secondary neurochemical events. The concept of ESCI and the underlying mechanisms have profound implications for the understanding of chronic traumatic encephalopathy (CTE) etiology because it has previously been suggested that repetitive axonal injury may be the primary CTE pathogenesis in susceptible individuals and it is best correlated with lifetime brain trauma load. Taken together, it appears that susceptibility to brain injury and downstream neurodegenerative diseases, such as CTE, can be conceptualized as a continuum of brain resilience. At one end is optimal resilience, capable of launching effective responses to injury with spontaneous recovery, and at the other end is diminished resilience with a compromised ability to respond and/or heal appropriately. Modulating factors such as one's total cumulative and synergistic brain trauma load, bioavailability of key nutrients needed for proper functioning of restorative metabolic pathways (specifically those involved in the deactivation and clearance of metabolic by-products of brain injury) are key to ultimately determining one's brain resilience.


Effect of acupuncture on post-hemorrhoidectomy pain: a randomized controlled trial.
To observe the clinical efficacy and safety of electroacupuncture (EA) in relieving pain after hemorrhoidectomy treatment for mixed hemorrhoids. This was a randomized controlled trial. We conducted a single-center, single-blind, and randomized controlled clinical trial. Seventy-two patients with mixed hemorrhoids who had undergone hemorrhoidectomy were randomly assigned to the following 2 groups: the EA treatment group (EA) received surround needling with EA (n=36), and the control group received sham acupuncture (SA) treatment (n=36). The treatment was conducted within 15 min after the completion of the surgery and lasted for 30 min. The pain intensity was recorded by using the visual analog scale as the primary outcome. Secondary outcomes were verbal rating scale and Wong-Baker Faces Pain Rating. These measurements were evaluated at 11 time points: once every hour in the first 8 h after the treatment, 24 and 48 h after the treatment, and at the first defecation. Besides, quality of life was measured by Symptom Checklist-90 Scale at 24 and 48 h follow-ups. The EA group had significantly lower visual analog scale scores at the 3 time points of 6, 24 h, and during the defecation (p<0.05). Verbal rating scale showed a significantly lower score in the treatment group compared to the SA group at 4 h after the treatment as well as during defecation (p<0.05). The Wong-Baker Faces Pain Rating scores of EA group were significantly lower at 5, 7, and 8 h after treatment and during defecation (p<0.05) compared with those of SA group. Acupuncture is effective in alleviating postoperative pain in patients who have undergone hemorrhoidectomy.


The effect of the UP4FUN pilot intervention on objectively measured sedentary time and physical activity in 10-12 year old children in Belgium: the ENERGY-project.
BAKCKGROUND: The first aim was to examine the effect of the UP4FUN pilot intervention on children's total sedentary time. The second aim was to investigate if the intervention had an effect on children's physical activity (PA) level. Finally, we aimed to investigate demographic differences (i.e. age, gender, ethnicity, living status and having siblings) between children in the intervention group who improved in sedentary time and PA at post-test and children in the intervention group who worsened in sedentary time and PA at post-test. The six weeks UP4FUN intervention was tested in a randomized controlled trial with pre-test post-test design with five intervention and five control schools in Belgium and included children of the 5th and 6th grade. The children wore accelerometers for seven days at pre- and post-test. Analyses included children with valid accelerometer data for at least two weekdays with minimum 10h-wearing time and one weekend day with 8h-wearing time. Final analyses included 372 children (60% girls, mean age = 10.9 ± 0.7 years). There were no significant differences in the change in sedentary time or light PA between intervention and control schools for the total sample or for the subgroup analyses by gender. However, children (specifically girls) in the intervention group had a higher decrease in moderate-to-vigorous PA than children in the control group. In the intervention group, children who lived with both parents and children with one or more siblings were less likely to reduce sedentary time after exposure to the intervention. Older children, girls and children who lived with both parents were less likely to increase light PA after the intervention. The UP4FUN intervention did not result in an effect on children's sedentary time. Based on the high amounts of accelerometer-derived sedentary time in this age group, more efforts are needed to develop strategies to reduce children's sedentary time.


Inpatient studies of a Kalman-filter-based predictive pump shutoff algorithm.
An insulin pump shutoff system can prevent nocturnal hypoglycemia and is a first step on the pathway toward a closed-loop artificial pancreas. In previous pump shutoff studies using a voting algorithm and a 1 min continuous glucose monitor (CGM), 80% of induced hypoglycemic events were prevented. The pump shutoff algorithm used in previous studies was revised to a single Kalman filter to reduce complexity, incorporate CGMs with different sample times, handle sensor signal dropouts, and enforce safety constraints on the allowable pump shutoff time. Retrospective testing of the new algorithm on previous clinical data sets indicated that, for the four cases where the previous algorithm failed (minimum reference glucose less than 60 mg/dl), the mean suspension start time was 30 min earlier than the previous algorithm. Inpatient studies of the new algorithm have been conducted on 16 subjects. The algorithm prevented hypoglycemia in 73% of subjects. Suspension-induced hyperglycemia is not assessed, because this study forced excessive basal insulin infusion rates. The new algorithm functioned well and is flexible enough to handle variable sensor sample times and sensor dropouts. It also provides a framework for handling sensor signal attenuations, which can be challenging, particularly when they occur overnight.


Dietary xylitol protects against weakening of bone biomechanical properties in ovariectomized rats.
The effects of dietary xylitol (xyl) on bone biomechanical properties in ovariectomized rats (ovx) were studied. Forty-two 3-mo-old female Wistar rats were divided into three groups of 14. Rats in two groups were ovariectomized, while those in the control group underwent a sham operation. All rats received a basal diet, and half of the ovx were given an additional 10 g/100 g dietary xyl supplementation. Three months later the rats were killed and their tibias, femurs and humeri were prepared. The tibias were used for analyses of bone density and trabecular bone volume (BV/TV) and for the three-point bending test. The femurs were used for the torsion test and for the loading test of the femoral neck. The humeri were used for analyses of bone ash weight and bone concentrations of calcium and phosphorus. Dietary xyl gave a significant protection against ovariectomy-caused decline of tibial stress in the three-point bending test, of femoral shear stress in the torsion test, and of stress of the femoral neck, without affecting bone elasticity values. Xyl restricted the ovariectomy-caused reduction in bone density, in bone ash weight and in concentrations of bone calcium and phosphorus. Furthermore, trabecular bone loss in ovx was significantly suppressed by dietary xyl. These results indicate that a 10% dietary supplementation of xyl in ovx has a protective effect against the weakening of bone biomechanical properties. This is related to greater BV/TV and maintained bone mineral content.


Dialects in the alarm calls of black- tailed prairie dogs (Cynomys ludovicianus): A case of cultural diffusion?
Prairie dogs (Cynomys spp.) produce an alarm call when a predator appears. Gunnison's prairie dogs (Cynomys gunnisoni) produce calls that vary in spectral structure according to predator type and specific characteristics unique to the individual predator, such as color and shape. These calls vary depending on geographic location and have been characterized as dialects. Black-tailed prairie dogs (Cynomys ludovicianus) also encode information about terrestrial predators in their alarm calls. However, nothing is known about whether there are dialects in these alarm calls, and if so, why these dialects might have arisen. We studied the alarm calls and habitat characteristics of eight different black-tailed prairie dog colonies within the Southwestern USA: Maxwell National Wildlife Refuge, NM, Vermejo Park Ranch, NM, Kiowa National Grasslands, NM, Rita Blanca National Grasslands, TX, and Comanche National Grasslands, CO. An analysis of time and frequency changes in alarm calls showed that colony sites that were located within 30 km each other did not have significant differences in alarm call structure, but sites that were more distant were significantly different from the other sites, consistent with the existence of dialects as well as with genetic models of isolation by distance. The acoustic structure of the alarm calls was not explained by differences in habitat characteristics at each site. A comparison of Squared Euclidian Distances (D2) of alarm calls versus linear distances between sites showed a significant positive correlation, implying that alarm call dialect differences increase the farther one site is from another. This relationship is consistent with a cultural diffusion model of dialects, and is similar to that observed in human dialects.


TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity.
Retinoic-acid-inducible gene-I (RIG-I; also called DDX58) is a cytosolic viral RNA receptor that interacts with MAVS (also called VISA, IPS-1 or Cardif) to induce type I interferon-mediated host protective innate immunity against viral infection. Furthermore, members of the tripartite motif (TRIM) protein family, which contain a cluster of a RING-finger domain, a B box/coiled-coil domain and a SPRY domain, are involved in various cellular processes, including cell proliferation and antiviral activity. Here we report that the amino-terminal caspase recruitment domains (CARDs) of RIG-I undergo robust ubiquitination induced by TRIM25 in mammalian cells. The carboxy-terminal SPRY domain of TRIM25 interacts with the N-terminal CARDs of RIG-I; this interaction effectively delivers the Lys 63-linked ubiquitin moiety to the N-terminal CARDs of RIG-I, resulting in a marked increase in RIG-I downstream signalling activity. The Lys 172 residue of RIG-I is critical for efficient TRIM25-mediated ubiquitination and for MAVS binding, as well as the ability of RIG-I to induce antiviral signal transduction. Furthermore, gene targeting demonstrates that TRIM25 is essential not only for RIG-I ubiquitination but also for RIG-I-mediated interferon- production and antiviral activity in response to RNA virus infection. Thus, we demonstrate that TRIM25 E3 ubiquitin ligase induces the Lys 63-linked ubiquitination of RIG-I, which is crucial for the cytosolic RIG-I signalling pathway to elicit host antiviral innate immunity.


Interdomain dynamics and ligand binding: molecular dynamics simulations of glutamine binding protein.
Periplasmic binding proteins from Gram-negative bacteria possess a common architecture, comprised of two domains linked by a hinge region, a fold which they share with the neurotransmitter-binding domains of ionotropic glutamate receptors (GluRs). Glutamine-binding protein (GlnBP) is one such protein, whose crystal structure has been solved in both open and closed forms. Multi-nanosecond molecular dynamics simulations have been used to explore motions about the hinge region and how they are altered by ligand binding. Glutamine binding is seen to significantly reduce inter-domain motions about the hinge region. Essential dynamics analysis of inter-domain motion revealed the presence of both hinge-bending and twisting motions, as has been reported for a related sugar-binding protein. Significantly, the influence of the ligand on GlnBP dynamics is similar to that previously observed in simulations of rat glutamate receptor (GluR2) ligand-binding domain. The essential dynamics analysis of GlnBP also revealed a third class of motion which suggests a mechanism for signal transmission in GluRs.


Global gene expression analysis reveals reduced abundance of putative microRNA targets in human prostate tumours.
Recently, microRNAs (miRNAs) have taken centre stage in the field of human molecular oncology. Several studies have shown that miRNA profiling analyses offer new possibilities in cancer classification, diagnosis and prognosis. However, the function of miRNAs that are dysregulated in tumours remains largely a mystery. Global analysis of miRNA-target gene expression has helped illuminate the role of miRNAs in developmental gene expression programs, but such an approach has not been reported in cancer transcriptomics. In this study, we globally analysed the expression patterns of miRNA target genes in prostate cancer by using several public microarray datasets. Intriguingly, we found that, in contrast to global mRNA transcript levels, putative miRNA targets showed a reduced abundance in prostate tumours relative to benign prostate tissue. Additionally, the down-regulation of these miRNA targets positively correlated with the number of types of miRNA target-sites in the 3' untranslated regions of these targets. Further investigation revealed that the globally low expression was mainly driven by the targets of 36 specific miRNAs that were reported to be up-regulated in prostate cancer by a miRNA expression profiling study. We also found that the transcript levels of miRNA targets were lower in androgen-independent prostate cancer than in androgen-dependent prostate cancer. Moreover, when the global analysis was extended to four other cancers, significant differences in transcript levels between miRNA targets and total mRNA backgrounds were found. Global gene expression analysis, along with further investigation, suggests that miRNA targets have a significantly reduced transcript abundance in prostate cancer, when compared with the combined pool of all mRNAs. The abnormal expression pattern of miRNA targets in human cancer could be a common feature of the human cancer transcriptome. Our study may help to shed new light on the functional roles of miRNAs in cancer transcriptomics.


Predictive Validity of ICD-11 PTSD as Measured by the Impact of Event Scale-Revised: A 15-Year Prospective Study of Political Prisoners.
The 11th edition of the International Classification of Diseases (ICD-11; World Health Organization, 2017) proposes a model of posttraumatic stress disorder (PTSD) that includes 6 symptoms. This study assessed the ability of a classification-independent measure of posttraumatic stress symptoms, the Impact of Event Scale-Revised (Weiss & Marmar, 1996), to capture the ICD-11 model of PTSD. The current study also provided the first assessment of the predictive validity of ICD-11 PTSD. Former East German political prisoners were assessed in 1994 (N = 144) and in 2008-2009 (N = 88) on numerous psychological variables using self-report measures. Of the participants, 48.2% and 36.8% met probable diagnosis for ICD-11 PTSD at the first and second assessments, respectively. Confirmatory factor analysis supported the factorial validity of the 3-factor ICD-11 model of PTSD, as represented by items selected from the Impact of Event Scale-Revised. Hierarchical multiple regression analysis demonstrated that, controlling for sex, the symptom clusters of ICD-11 PTSD (reexperiencing, avoidance, and sense of threat) significantly contributed to the explanation of depression (R2 = .17), quality of life (R2 = .21), internalized anger (R2 = .10), externalized anger (R2 = .12), hatred of perpetrators (R2 = .15), dysfunctional disclosure (R2 = .27), and social acknowledgment as a victim (R2 = .12) across the 15-year study period. Current findings add support for the factorial and predictive validity of ICD-11 PTSD within a unique cohort of political prisoners.


Biochemical analysis of preliminary clusters in the non-lineage panel.
After the preliminary clustering of the whole workshop panel using flowcytometry data, we selected those clusters that were composed of mAbs that bound a broad range of cells. To obtain evidence that mAbs in each of these preliminary clusters detected the same antigen, they were tested for their capacity to compete with each other for binding to a target cell and the molecular weights of their antigens were estimated after immunoprecipitation. Most preliminary clusters in the non-lineage panel contained control mAbs that had been characterised in one of the previous workshops, and this study therefore increased the number of mAbs available to each of these non-lineage antigens. One new interesting cluster, BoWC14, was described which defined an antigen on myeloid cells and a subpopulation of CD4, CD8 and WC1 T lymphocytes (BT3/8.12 and IL-A155). An additional cluster, which did not fit into any other panel, contained mAbs specific for an antigen restricted to the erythroid lineage and received the nomenclature BoWC15 (ANA8, IL-A135, IL-A137, IL-A138 and IL-A160).


Effects of psilocybin therapy on personality structure.
To explore whether psilocybin with psychological support modulates personality parameters in patients suffering from treatment-resistant depression (TRD). Twenty patients with moderate or severe, unipolar, TRD received oral psilocybin (10 and 25 mg, one week apart) in a supportive setting. Personality was assessed at baseline and at 3-month follow-up using the Revised NEO Personality Inventory (NEO-PI-R), the subjective psilocybin experience with Altered State of Consciousness (ASC) scale, and depressive symptoms with QIDS-SR16. Neuroticism scores significantly decreased while Extraversion increased following psilocybin therapy. These changes were in the direction of the normative NEO-PI-R data and were both predicted, in an exploratory analysis, by the degree of insightfulness experienced during the psilocybin session. Openness scores also significantly increased following psilocybin, whereas Conscientiousness showed trend-level increases, and Agreeableness did not change. Our observation of changes in personality measures after psilocybin therapy was mostly consistent with reports of personality change in relation to conventional antidepressant treatment, although the pronounced increases in Extraversion and Openness might constitute an effect more specific to psychedelic therapy. This needs further exploration in future controlled studies, as do the brain mechanisms of postpsychedelic personality change.


Avicenna's doctrine about arterial hypertension.
Arterial hypertension is a major risk factor for the development of cardiovascular diseases. Data from observational studies indicate that it may affect 90% of the general population during their lifetime. Despite much research that has been done, the exact cause of this disorder is still unknown. Avicenna (Ibn Sina) in his masterpiece The Canon of Medicine described most of the clinical features, causes, and complications which are consistent with hypertension symptoms based on modern medicine. He described in detail the symptoms of hypertension such as headache, heaviness in the head, sluggish movements, general redness and warm to touch feel of the body, prominent, distended and tense veins, fullness of the pulse, distension of the skin, coloured and dense urine, loss of appetite, weak eye sight, impairment of thinking, yawning, and drowsiness. Moreover, Avicenna described haemorrhage and sudden death as the complications of hypertension. Due to the importance of this issue, we wanted to call the reader's attention to Avicenna's views about what corresponds to hypertension in modern medicine.


Altered cannabinoid CB1 receptor mRNA expression in mesencephalon from mice exposed to repeated methamphetamine and methanandamide treatments.
Since among others also our previous studies suggested an interaction between the endocannabinoid system and methamphetamine brain mechanisms we focused on possible changes in relative expression of cannabinoid CB1 receptor mRNA in mesencephalon from mice sensitized by repeated treatments to methamphetamine stimulatory effects and cross-sensitized by cannabinoid CB1 receptor agonist methanandamide pre-treatment. The Open Field Test was used to measure changes in terms of behavioural sensitization or cross-sensitization to drug effects on locomotion in male mice treated repeatedly with either methamphetamine or methamphetamine after pre-treatment with methanandamide. After each measurement one third of animals were sacrificed and the brain was stored. RNA was isolated from the midbrain and used for reverse transcription and subsequent real-time PCR. The evaluation of behavioural drug effects showed both development of sensitization to methamphetamine stimulatory effects after repeated treatment and cross-sensitization to them by pre-treatment with cannabinoid receptor CB1 agonist methanandamide. Real-time PCR analyses revealed an increase in CB1 receptor mRNA expression after the first dose of methanandamide followed by decrease after the combined treatment with methamphetamine challenge dose. Our findings suggest that particularly repeated pre-treatment with CB1 agonist methanandamide can elicit increase in the mRNA expression level at least in the mouse mesencephalon neurons associated with cross-sensitization to methamphetamine stimulatory effects.


Non-symbolic numerosities do not automatically activate spatial-numerical associations: Evidence from the SNARC effect.
The SNARC (spatial-numerical association of response codes) effect is the finding that people are generally faster to respond to smaller numbers with left-sided responses and larger numbers with right-sided responses. The SNARC effect has been widely reported for responses to symbolic representations of number such as digits. However, there is mixed evidence as to whether it occurs for non-symbolic representations of number, particularly when magnitude is irrelevant to the task. Mitchell et al. reported a SNARC effect when participants were asked to make orientation decisions to arrays of one-to-nine triangles (pointing upwards vs. pointing downwards) and concluded that SNARC effects occur for non-symbolic, non-canonical representations of number. They additionally reported that this effect was stronger in the subitising range. However, here we report four experiments that do not replicate either of these findings. Participants made upwards/inverted decisions to one-to-nine triangles where total surface area was either controlled across numerosities (Experiments 1, 2, and 4) or increased congruently with numerosity (Experiment 3). There was no evidence of a SNARC effect either across the full range or within the subset of the subitising range. The results of Experiment 4 (in which we presented the original stimuli of Mitchell et al.) suggested that visual properties of non-symbolic displays can prompt SNARC-like effects driven by visual cues rather than numerosity. Taken in the context of other recent findings, we argue that non-symbolic representations of number do not offer a direct and automatic route to numerical-spatial associations.


A general basis for quarter-power scaling in animals.
It has been known for decades that the metabolic rate of animals scales with body mass with an exponent that is almost always <1, >2/3, and often very close to 3/4. The 3/4 exponent emerges naturally from two models of resource distribution networks, radial explosion and hierarchically branched, which incorporate a minimum of specific details. Both models show that the exponent is 2/3 if velocity of flow remains constant, but can attain a maximum value of 3/4 if velocity scales with its maximum exponent, 1/12. Quarter-power scaling can arise even when there is no underlying fractality. The canonical "fourth dimension" in biological scaling relations can result from matching the velocity of flow through the network to the linear dimension of the terminal "service volume" where resources are consumed. These models have broad applicability for the optimal design of biological and engineered systems where energy, materials, or information are distributed from a single source.


All-terrain vehicle accidents at a level II trauma center in Indiana: an 8-year retrospective review.
This study profiles the all-terrain vehicle (ATV) accident victims who were admitted at a level II trauma center. We retrospectively reviewed the trauma registry data for 111 patients who were admitted to the Parkview Hospital Level II Trauma Center following ATV crashes between January 1996 and June 2003. Forty-four percent of the patients were < or =16 years of age, and the average age overall was 22.49 years. Most of the 111 victims were men, and only 18.9% of those injured were wearing helmets. Although they only make up 14% of the driver population, children <16 years of age account for almost 40% of all ATV-related injuries and >35% of all ATV-related deaths in the United States. Helmets can reduce the risk of head injury, but only 21 states have helmet laws. Safety legislation should be adopted and would likely save lives and prevent injuries.


Activation of endogenous p53 by combined p19Arf gene transfer and nutlin-3 drug treatment modalities in the murine cell lines B16 and C6.
Reactivation of p53 by either gene transfer or pharmacologic approaches may compensate for loss of p19Arf or excess mdm2 expression, common events in melanoma and glioma. In our previous work, we constructed the pCLPG retroviral vector where transgene expression is controlled by p53 through a p53-responsive promoter. The use of this vector to introduce p19Arf into tumor cells that harbor p53wt should yield viral expression of p19Arf which, in turn, would activate the endogenous p53 and result in enhanced vector expression and tumor suppression. Since nutlin-3 can activate p53 by blocking its interaction with mdm2, we explored the possibility that the combination of p19Arf gene transfer and nutlin-3 drug treatment may provide an additive benefit in stimulating p53 function. B16 (mouse melanoma) and C6 (rat glioma) cell lines, which harbor p53wt, were transduced with pCLPGp19 and these were additionally treated with nutlin-3 or the DNA damaging agent, doxorubicin. Viral expression was confirmed by Western, Northern and immunofluorescence assays. p53 function was assessed by reporter gene activity provided by a p53-responsive construct. Alterations in proliferation and viability were measured by colony formation, growth curve, cell cycle and MTT assays. In an animal model, B16 cells were treated with the pCLPGp19 virus and/or drugs before subcutaneous injection in C57BL/6 mice, observation of tumor progression and histopathologic analyses. Here we show that the functional activation of endogenous p53wt in B16 was particularly challenging, but accomplished when combined gene transfer and drug treatments were applied, resulting in increased transactivation by p53, marked cell cycle alteration and reduced viability in culture. In an animal model, B16 cells treated with both p19Arf and nutlin-3 yielded increased necrosis and decreased BrdU marking. In comparison, C6 cells were quite susceptible to either treatment, yet p53 was further activated by the combination of p19Arf and nutlin-3. To the best of our knowledge, this is the first study to apply both p19Arf and nutlin-3 for the stimulation of p53 activity. These results support the notion that a p53 responsive vector may prove to be an interesting gene transfer tool, especially when combined with p53-activating agents, for the treatment of tumors that retain wild-type p53.


The contribution of the C-824T tyrosine hydroxylase polymorphism to the prevalence of hypertension in a South African cohort: the SABPA study.
The C-824T single nucleotide polymorphism in the promoter region of the tyrosine hydroxylase gene has been associated with hypertension. It is well documented that African South Africans exhibit a higher prevalence of hypertension than Caucasians. However, the possible role of this mutation on 24-hour ambulatory blood pressure (AMBP) has not been investigated in African South Africans. Blood samples of 409 South Africans were screened for the mutation. Ambulatory blood pressure and lifestyle factors were also measured. Africans had higher incidence of hypertension and higher occurrence of the mutation. However, the contribution of the tyrosine hydroxylase C-824T single nucleotide polymorphism to hypertension could not be confirmed in our cohort.


Identification of patients with acute myeloblastic leukemia who benefit from the addition of gemtuzumab ozogamicin: results of the MRC AML15 trial.
Antibody-directed chemotherapy for acute myeloid leukemia (AML) may permit more treatment to be administered without escalating toxicity. Gemtuzumab ozogamicin (GO) is an immunoconjugate between CD33 and calicheamicin that is internalized when binding to the epitope. We previously established that it is feasible to combine GO with conventional chemotherapy. We now report a large randomized trial testing the addition of GO to induction and/or consolidation chemotherapy in untreated younger patients. In this open-label trial, 1,113 patients, predominantly younger than age 60 years, were randomly assigned to receive a single dose of GO (3 mg/m(2)) on day 1 of induction course 1 with one of the following three induction schedules: daunorubicin and cytarabine; cytarabine, daunorubicin, and etoposide; or fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin. In remission, 948 patients were randomly assigned to GO in course 3 in combination with amsacrine, cytarabine, and etoposide or high-dose cytarabine. The primary end points were response rate and survival. The addition of GO was well tolerated with no significant increase in toxicity. There was no overall difference in response or survival in either induction of consolidation. However, a predefined analysis by cytogenetics showed highly significant interaction with induction GO (P = .001), with significant survival benefit for patients with favorable cytogenetics, no benefit for patients with poor-risk disease, and a trend for benefit in intermediate-risk patients. An internally validated prognostic index identified approximately 70% of patients with a predicted benefit of 10% in 5-year survival. A substantial proportion of younger patients with AML have improved survival with the addition of GO to induction chemotherapy with little additional toxicity.


Silent Cerebral Small Vessel Disease in Restless Legs Syndrome.
Growing literature suggests that patients with restless legs syndrome (RLS) may be at increased risk for hypertension, heart disease, and stroke. Cerebral small vessel disease (SVD) is a known risk factor for clinical stroke. This study evaluated silent cerebral SVD by MRI in patients with RLS, in the absence of a history of previous clinical stroke or known stroke risk factors and taking into account disease duration. Fifty-three patients with RLS < 10 y were prospectively recruited along with 44 with RLS > 10 y and 74 normal controls. A magnetic resonance imaging study was obtained from all subjects and scans were analyzed for area and volume of SVD. There was a significant increase in SVD area in the entire group of RLS patients compared to controls (P = 0.036); this was almost entirely driven by the group with RLS > 10 y. SVD area and volume were significantly increased in patients with RLS > 10 y with respect to both controls (P < 0.0001 and P < 0.0014, respectively) and RLS < 10 y (P < 0.00022 and P < 0.003, respectively). Age, duration of RLS, and the interaction of age and duration of RLS were independent predictors of SVD disease. Duration of RLS was an independent predictor of the burden of cerebral SVD (area P < 0.00012 and volume P < 0.0025), whereas sex and insomnia were not. RLS duration should be taken into account when analyzing the association between RLS and cerebrovascular disease; our data support the hypothesis that a long-lasting RLS and its accompanying periodic limb movements in sleep are a risk factor for silent SVD and perhaps for the development of clinical stroke.


The effect of single motherhood on smoking by socioeconomic status and race/ethnicity.
We examined the association between parenting young children and smoking among US single women compared with married women, and whether this effect is moderated by socioeconomic status and race/ethnicity. Our main finding is that having children reduces smoking except among single white women, and women with low income. We used the Tobacco Use Supplement of the Current Population Survey, a nationally representative dataset (1995-96, n=70,019). Log-binomial regression analysis was used to estimate the association between parenting responsibility (i.e., presence of children aged 0-4 and 5-17) and daily smoking status, after taking into consideration marital status, income, and race/ethnicity. Single women faced a higher risk of smoking than married women. Parenting was protective against smoking among married women but not among single women. Additionally, among single women, the associations between parenting and smoking varied by income and race/ethnicity. Parenting increased the risk of smoking among single women in the lowest income quartile. Single black and Hispanic women with children had a risk of smoking similar to that of their childless counterparts. However, single white women with children were more likely to smoke than their childless counterparts. Smoking cessation interventions and programs to reduce environmental tobacco smoke should recognize that the co-occurrence of single motherhood, parenting responsibility and low-income may increase the risk of smoking. This is particularly significant given the rapid growth of the single women population, and their concentration in poverty in the USA. The finding that parenting is protective against smoking among single minority women, who presumably experience significant stressors, calls for a more thorough investigation of smoking behavior among minority women, and suggests the importance of stress buffers such as social support.


First Report of Pepper vein yellows virus Infecting Sweet Pepper in Spain.
In November 2012, unusual symptoms were observed in plants of sweet pepper (Capsicum annuum L.) grown in commercial greenhouses of Almería Province, southeastern Spain. Symptoms included interveinal yellowing, upward leaf curling, and internode shortening, and were more evident in the upper part of the plant. Abnormal ripening of fruits was observed in symptomatic plants, with fruits remaining orange in the red varieties and yellow in the orange varieties, thus reducing their marketability. During December 2012 and January 2013, severe outbreaks of this disease syndrome occurred, with many greenhouses exhibiting almost 100% incidence. The symptoms observed were similar to those reported for isolates of Pepper vein yellows virus (PeVYV, genus Polerovirus, family Luteoviridae) (previously also named Pepper yellow leaf curl virus [PYLCV] and Pepper yellows virus [PYV]) (2,4). Twenty five symptomatic leaf and/or fruit samples (some of them supplied by Zeraim Ibérica, S.A.), each from a different greenhouse, were analyzed and all reacted positively in double-antibody sandwich-ELISA with an antiserum against the polerovirus Cucurbit aphid-borne yellows virus (CABYV) (Sediag, Longvic, France), known to cross-react with PeVYV (2). Total RNA was extracted by TRIsure reagent (Bioline, London, United Kingdom) from symptomatic leaves and analyzed by reverse transcription (RT)-PCR with primers Pol-G-F (5'-GAYTGCTCYGGYTTYGACTGGAG-3') and Pol-G-R (5'-GATYTTATAYTCATGGTAGGCCTTGAG-3') designed for universal detection of poleroviruses by amplifying the RNA-dependent RNA polymerase (RdRp) and coat protein (CP) partial genes (3). DNA fragments of the expected size (1.1 kbp) were amplified supporting a polerovirus infection in all the analyzed samples. The PCR product obtained from one sample (Almería-1) was extracted from agarose gel with a QIAquick gel extraction kit (Qiagen, Hilden, Germany), cloned in pGEM-T Easy vector (Promega, Madison, WI), and one clone was sequenced (Macrogen Inc., Seoul, South Korea). The PCR products amplified from three other samples (2-13, 7-13, and 8-13) were directly sequenced. The nucleotide identity between the amplified fragments (GenBank Accession Nos. KC769487, KC839992 to 94), calculated after alignment with ClustalW, was 99.7 to 100%. The highest nucleotide identity of the Spanish sequences was with a PeVYV isolate from Turkey (FN600344, named as PYV) (98.5 to 98.7%). The spread of PeVYV in Spain is additional evidence of the emergence of this virus as a global threat for pepper crops after its first detection in Japan in 1995 and recent reports from the Mediterranean Basin (1,2). References: (1) N. Buzkan et al. Arch. Virol. 158:881, 2013. (2) A. Dombrovsky et al. Phytoparasitica 38:477, 2010. (3) D. Knierim et al. Plant Pathol. 59:991, 2010. (4) R. Murakami et al. Arch. Virol. 156:921, 2011.


The "Strengthen your ankle" program to prevent recurrent injuries: A randomized controlled trial aimed at long-term effectiveness.
Recurrent ankle sprains can be reduced by a neuromuscular training program (NMT). The way NMT is delivered may influence the incidence of long term recurrent injuries, residual pain and disability. This RCT with a follow-up of twelve months, evaluated whether the implementation method of a proven effective NMT program delivered by a mobile application or a written instruction booklet, resulted in differences in injury incidence rates, functional ankle disability/pain in the long term, assuming equal compliance - as is shown in previous research - with the 8-week intervention. 220 athletes with a history of ankle sprain were recruited for this RCT. 110 athletes were offered the freely available "Strengthen your ankle App" and the other 110 received a printed Booklet. Primary outcome measure was incidence density of ankle sprains. Secondary outcome measures were residual pain/disability and the individual cumulative number of ankle sprains during follow-up. The incidence densities of self-reported ankle sprain recurrences were not significantly different between both groups (HR 1.06; 95% CI 0.76-1.49). Median FADI (Functional Ankle and Disability Index) scores increased equally over time in both groups, indicating a lower rate of limitation and pain in both groups at follow-up. Neither FADI scores nor cumulative recurrent injuries were significantly different between groups. This study showed that the implementation method of a NMT program by using an App or a Booklet did neither lead to different injury incidence rates in the long term nor did it influence residual functional disability/pain. Assuming equal compliance during the 8-week intervention, both methods show similar effectiveness in twelve-month follow-up.


Specific incremental test in elite squash players.
To compare cardiorespiratory responses between incremental treadmill (non-specific) and field (sport specific) tests in elite squash players. Seven elite players (ranked 1 to 25 in their national federation including the World number 1) randomly performed an incremental treadmill test (TT) and a squash specific graded test (ST) to exhaustion. The ST consisted of repeated displacements replicating the game of squash, at increasing speed on the court. In both tests, ventilatory variables and heart rate were determined at the ventilatory threshold, respiratory compensation point, and maximal loads (max). Heart rate and percentage maximal oxygen uptake (VO2MAX) at the ventilatory threshold and respiratory compensation point were not different between the ST and TT, whereas VO2MAX was higher in the ST than in the TT (63.6 (3.0) v 54.9 (2.5) ml/kg/min; p < 0.001). Time to exhaustion was not different between the ST and TT (1056 (180) v 962 (71) seconds) but correlated with the ranking of the players only in the ST (r = -0.96, p < 0.001). VO2MAX values derived from laboratory testing were not relevant for accurately estimating fitness in elite squash players. So the ST may be used as an additional test for determination of training intensity. Improved training advice for prescribing aerobic exercise or perfecting stroke technique may result from these results.


Correlation of cardiac output and sevoflurane required to maintain anesthetic depth targeted with entropy index.
We investigated the correlation of reduced cardiac output on required sevoflurane to maintain targeted anesthesia depth. 36 patients undergoing coronary artery bypass grafting with cardiopulmonary bypass were prospectively included in the study. Inspired sevoflurane concentration was adjusted to ensure state entropy index < 40. Analgesia was provided by either boluses of fentanyl 200 μg or continuous infusion of fentanyl 5 μg/kg/h; the total dose of fentanyl administered in the patients was not different (fentanyl boluses 6.5 ± 0.3 μg/kg/h vs. fentanyl infusion 5 μg/kg/h). Cardiac-index (CI), end tidal sevoflurane (ETsev) and entropy index were measured simultaneously at 1-5 min after sternotomy, during internal mammary artery harvesting and during pericardiotomy. 108 sets of variables (entropy index, ETsev, CI) were recorded from 36 subjects at three time points; 13 sets were excluded due to technical drawbacks in measurements. 95 data sets were eligible for analysis. Sixty-five data sets measured in patients with target state entropy index were analyzed to establish the relationship between CI and ETsev. We did not find a linear correlation between ETsev and CI in patients with target entropy index (correlation coefficient = 0.18, P = 0.14). The ETsev necessary to maintain the target level of anesthesia was lower in patients with CI ≤ 2.2 l/min/m2 (1.15% ± 0.28%) than patients with CI > 2.2 l/min/m2 (1.37% ± 0.31%), P = 0.01. Relationship between CI and ETsev required for maintaining target level of anesthesia is non-linear. Patients with CI ≤ 2.2 l/min/m2 need lower levels of the ETsev for maintenance of the target anesthesia at an entropy index < 40.


Mood and substance use disorders among adults seeking speech treatment for stuttering.
Stuttering has been associated with a range of anxiety disorders, including social phobia. In the general community, anxiety disorders are frequently associated with increased rates of mood and substance use disorders. Therefore, in the present study, the authors sought to determine the rate of mood and substance use disorders among adults who stutter. The study employed a matched case-control design. Participants included 92 adults seeking treatment for stuttering and 920 age- and gender-matched controls. Mental health assessments were conducted via a computerized psychiatric diagnostic interview. Conditional logistic regression models were used to estimate odds ratios for the prevalence of mood and substance use disorders in both groups. When compared with matched controls, adults seeking treatment for stuttering had twofold increased odds of meeting criteria for a 12-month diagnosis of any mood disorder but were not found to report significantly higher lifetime prevalence rates for any substance use disorder. Although adults who stutter in the present study were characterized by significantly higher rates of mood disorders than matched controls, they do not appear to self-medicate with substances such as alcohol. Results are discussed in terms of treatment implications and possible reasons why adults who stutter may avoid alcohol.


Analytical model of spectrometer-based two-beam spectral interferometry.
We report an analytical model of signal formation in spectrometer-based two-beam spectral interferometry. Considering the pixel size, the optical resolution and the spectral resolution of the spectrometer, and dispersion, the model represents the signal recorded by a spectrometer based on a diffraction grating and linear detector array. The model is general, but degenerates to more familiar forms with simplifying assumptions. The model is validated by comparison with experimental measurements, where it is shown that the model can accurately predict both signal fall-off and axial resolution for Fourier-domain optical coherence tomography imaging. The model may be useful for determining design specifications and expected performance parameters for spectrometers for spectral interferometry.


Inhibition of SARS-CoV-2 (previously 2019-nCoV) infection by a highly potent pan-coronavirus fusion inhibitor targeting its spike protein that harbors a high capacity to mediate membrane fusion.
The recent outbreak of coronavirus disease (COVID-19) caused by SARS-CoV-2 infection in Wuhan, China has posed a serious threat to global public health. To develop specific anti-coronavirus therapeutics and prophylactics, the molecular mechanism that underlies viral infection must first be defined. Therefore, we herein established a SARS-CoV-2 spike (S) protein-mediated cell-cell fusion assay and found that SARS-CoV-2 showed a superior plasma membrane fusion capacity compared to that of SARS-CoV. We solved the X-ray crystal structure of six-helical bundle (6-HB) core of the HR1 and HR2 domains in the SARS-CoV-2 S protein S2 subunit, revealing that several mutated amino acid residues in the HR1 domain may be associated with enhanced interactions with the HR2 domain. We previously developed a pan-coronavirus fusion inhibitor, EK1, which targeted the HR1 domain and could inhibit infection by divergent human coronaviruses tested, including SARS-CoV and MERS-CoV. Here we generated a series of lipopeptides derived from EK1 and found that EK1C4 was the most potent fusion inhibitor against SARS-CoV-2 S protein-mediated membrane fusion and pseudovirus infection with IC50s of 1.3 and 15.8 nM, about 241- and 149-fold more potent than the original EK1 peptide, respectively. EK1C4 was also highly effective against membrane fusion and infection of other human coronavirus pseudoviruses tested, including SARS-CoV and MERS-CoV, as well as SARSr-CoVs, and potently inhibited the replication of 5 live human coronaviruses examined, including SARS-CoV-2. Intranasal application of EK1C4 before or after challenge with HCoV-OC43 protected mice from infection, suggesting that EK1C4 could be used for prevention and treatment of infection by the currently circulating SARS-CoV-2 and other emerging SARSr-CoVs.


Serpentine channels: micro-rheometers for fluid relaxation times.
We propose a novel device capable of measuring relaxation times of viscoelastic fluids as small as 1 ms. In contrast to most rheometers, which by their very nature are concerned with producing viscometric or nearly-viscometric flows, here we make use of an elastic instability that occurs in the flow of viscoelastic fluids with curved streamlines. To calibrate the rheometer we combine simple scaling arguments with relaxation times obtained from first normal-stress difference data measured in a classical shear rheometer. As an additional check we also compare these relaxation times to those obtained from Zimm theory and good agreement is observed. Once calibrated, we show how the serpentine rheometer can be used to access smaller polymer concentrations and lower solvent viscosities where classical measurements become difficult or impossible to use due to inertial and/or resolution limitations. In the absence of calibration, the serpentine channel can still be a very useful comparative or index device.


Effect of method of delivering nicarbazin to mallards on plasma 4,4'-dinitrocarbanilide levels and reproduction.
Nicarbazin (NCZ), a coccidiostat used in the poultry industry, has been developed as a contraceptive for resident Canada geese. We tested the efficacy of NCZ as a contraceptive using mallards (Anas platyrhynchos) as a model for Canada geese. Nicarbazin-treated corn was fed ad libitum for 14 d at 0, 750, 1,000, or 1,500 ppm. Plasma and egg levels of 4,4'-dinitrocarbanilide (DNC), the active anticoccidial component of NCZ, differed among treatment groups in a dose-response relationship, but plasma levels did not differ between sexes. Nicarbazin caused a decrease in egg weight, but there was no effect of NCZ on the numbers of eggs laid per female per day. Nicarbazin did not significantly impact bird health. An additional trial tested the effect of the method of NCZ delivery on plasma DNC levels. Mallards were given NCZ daily for 12 d either by gavage with a corn oil suspension, gavage with a water suspension, peroral administration of a capsule, or feeding 500 mg of NCZ/kg of pelleted feed ad libitum. The method of delivery significantly affected plasma DNC levels, with the highest levels in the corn oil suspension group and the lowest levels in the pelleted feed group. This is likely due to decreased availability of NCZ in a pellet compared with gavage with a suspension or capsule. Mallards receiving 34.2 mg of NCZ/kg of BW when fed cracked corn coated with NCZ daily for 14 d had higher plasma DNC levels than those obtained by liquid gavage, capsule, or pelleted NCZ feed. For maximum effect in the field, NCZ should be coated onto corn. A higher concentration of NCZ is needed in pelleted feed to obtain comparable plasma DNC levels to allow for the decreased absorption of DNC.


Basigin is a receptor essential for erythrocyte invasion by Plasmodium falciparum.
Erythrocyte invasion by Plasmodium falciparum is central to the pathogenesis of malaria. Invasion requires a series of extracellular recognition events between erythrocyte receptors and ligands on the merozoite, the invasive form of the parasite. None of the few known receptor-ligand interactions involved are required in all parasite strains, indicating that the parasite is able to access multiple redundant invasion pathways. Here, we show that we have identified a receptor-ligand pair that is essential for erythrocyte invasion in all tested P. falciparum strains. By systematically screening a library of erythrocyte proteins, we have found that the Ok blood group antigen, basigin, is a receptor for PfRh5, a parasite ligand that is essential for blood stage growth. Erythrocyte invasion was potently inhibited by soluble basigin or by basigin knockdown, and invasion could be completely blocked using low concentrations of anti-basigin antibodies; importantly, these effects were observed across all laboratory-adapted and field strains tested. Furthermore, Ok(a-) erythrocytes, which express a basigin variant that has a weaker binding affinity for PfRh5, had reduced invasion efficiencies. Our discovery of a cross-strain dependency on a single extracellular receptor-ligand pair for erythrocyte invasion by P. falciparum provides a focus for new anti-malarial therapies.


Oxidation of methane by an N-bridged high-valent diiron-oxo species: electronic structure implications on the reactivity.
High-valent iron-oxo species are key intermediates in C-H bond activation of several substrates including alkanes. The biomimic heme and non-heme mononuclear Fe(IV)=O complexes are very popular in this area and have been thoroughly studied over the years. These species despite possessing aggressive catalytic ability, cannot easily activate inert C-H bonds such as those of methane. In this context dinuclear complexes have gained attention, particularly μ-nitrido dinuclear iron species [(TPP)(m-CBA)Fe(IV)(μ-N)Fe(IV)(O)(TPP(˙+))](-) reported lately exhibits remarkable catalytic abilities towards substrates such as methane. Here using DFT methods, we have explored the electronic structure and complex spin-state energetics present in this species. To gain insights into the nature of bonding, we have computed the absorption, the EPR and the Mössbauer parameters and have probed the mechanism of methane oxidation by the dinuclear Fe(IV)=O species. Calculated results are in agreement with the experimental data and our calculations predict that in [(TPP)(m-CBA)Fe(IV)(μ-N)Fe(IV)(O)(TPP(˙+))](-)species, the two high-spin iron centres are antiferromagnetically coupled leading to a doublet ground state. Our calculations estimate an extremely low kinetic barrier of 26.6 kJ mol(-1) (at doublet surface) for the C-H bond activation of methane by the dinuclear Fe(IV)=O species. Besides these mechanistic studies on the methane activation reveal the unique electronic cooperativity present in this type of dinuclear complex and unravel the key question of why mononuclear analogues are unable to perform such reactions.


NO-sulindac inhibits the hypoxia response of PC-3 prostate cancer cells via the Akt signalling pathway.
Nitric oxide-donating non-steroidal anti-inflammatory drugs are safer than traditional NSAIDs and inhibit the growth of prostate cancer cells with greater potency than NSAIDs. In vivo, prostate cancer deposits are found in a hypoxic environment which induces resistance to chemotherapy. The aim of this study was to assess the effects and mechanism of action of a NO-NSAID called NO-sulindac on the PC-3 prostate cancer cell line under hypoxic conditions. NO-sulindac was found to have pro-apoptotic, cytotoxic, and anti-invasive effect on PC-3 cells under normoxia and hypoxia. NO-sulindac was significantly more cytotoxic than sulindac at all oxygen levels. The sulindac/linker and NO-releasing subunits both contributed to the cytotoxic effects of NO-sulindac. Resistance of PC-3 cells to NO-sulindac was induced as the oxygen concentration declined. Hypoxia-induced chemoresistance was reversed by knocking-down hypoxia-inducible factor-1alpha (HIF-1alpha) mRNA using RNAi. Nuclear HIF-1alpha levels were upregulated at 0.2% oxygen but reduced by treatment with NO-sulindac, as was Akt phosphorylation. NO-sulindac treatment of hypoxic PC-3 cells transfected with a reporter construct, downregulated activation of the hypoxia response element (HRE) promoter. Co-transfection of PC-3 cells with the HRE promoter reporter construct and myr-Akt (constitutively active Akt) plasmids reversed the NO-sulindac induced reduction in HRE activation. Real-time polymerase chain reaction analysis of hypoxic, NO-sulindac treated PC-3 cells showed downregulation of lysyl oxidase and carbonic anhydrase IX mRNA expression. Collectively, these novel findings demonstrate that NO-sulindac directly inhibits the hypoxia response of PC-3 prostate cancer cells by inhibiting HIF-1alpha translation via the Akt signalling pathway. The ability of NO-sulindac to inhibit tumour adaption to hypoxia has considerable relevance to the future management of prostate cancer with the same cellular properties as PC-3.


MicroRNA-196a links human body fat distribution to adipose tissue extracellular matrix composition.
Abdominal fat mass is associated with metabolic risk whilst gluteal femoral fat is paradoxically protective. MicroRNAs are known to be necessary for adipose tissue formation and function but their role in regulating human fat distribution remains largely unexplored. An initial microarray screen of abdominal subcutaneous and gluteal adipose tissue, with validatory qPCR, identified microRNA-196a as being strongly differentially expressed between gluteal and abdominal subcutaneous adipose tissue. We found that rs11614913, a SNP within pre-miR-196a-2 at the HOXC locus, is an eQTL for miR-196a expression in abdominal subcutaneous adipose tissue (ASAT). Observations in large cohorts showed that rs11614913 increased waist-to-hip ratio, which was driven specifically by an expansion in ASAT. In further experiments, rs11614913 was associated with adipocyte size. Functional studies and transcriptomic profiling of miR-196a knock-down pre-adipocytes revealed a role for miR-196a in regulating pre-adipocyte proliferation and extracellular matrix pathways. These data identify a role for miR-196a in regulating human body fat distribution. FUND: This work was supported by the Medical Research Council and Novo Nordisk UK Research Foundation (G1001959) and Swedish Research Council. We acknowledge the OBB-NIHR Oxford Biomedical Research Centre and the British Heart Foundation (BHF) (RG/17/1/32663). Work performed at the MRC Epidemiology Unit was funded by the United Kingdom's Medical Research Council through grants MC_UU_12015/1, MC_PC_13046, MC_PC_13048 and MR/L00002/1.


Ex vivo selection for oncoretrovirally transduced green fluorescent protein-expressing CD34-enriched cells increases short-term engraftment of transduced cells in baboons.
In an effort to improve hematopoietic stem cell gene transfer rates using gibbon ape leukemia virus (GALV)-pseudotype retroviral vectors in baboons, we have studied preselection of transduced green fluorescent protein (GFP)-expressing CD34-enriched marrow cells. Three animals were transplanted with GFP-selected (GS) CD34-enriched marrow. To ensure engraftment, preselected GFP-positive cells were infused together with unselected neo-transduced cells. After transduction on fibronectin, cells were cultured for an additional 2 days to allow for expression of GFP. GFP-expressing cells were enriched by fluorescence-activated cell sorting and infused together with cells from the unselected fractions after myeloablative irradiation of the recipient. Three other animals were transplanted with GFP-transduced CD34-enriched cells without prior GFP selection (GU). At 4 weeks after transplant, the percentage of GFP-expressing white blood cells was significantly higher in the GS group (6.6%) than in the GU group (1.3%) (p < 0.002). The higher gene transfer levels in the animals transplanted with GS cells gradually declined, and by day 100 after transplant, gene transfer levels were similar in both groups. PCR analysis performed on genomic DNA isolated from peripheral blood cells demonstrated that the decline in GFP-positive cells was due to the loss of gene-marked cells and not due to loss of expression. These results show that transplantation of CD34-positive marrow cells selected for GFP-positive cells after transduction provides high levels of transduced granulocytes in the short term. However, using this experimental design with concomitant infusion of unselected cells and the use of oncoretroviral vectors, preenrichment of vector-expressing, transduced CD34-enriched cells does not improve long-term persistence and expression.


Healthcare issues of detainees in police custody in London, UK.
Little is known about the general healthcare needs of detainees in police custody. The aims of this study were to: determine the level of general health issues, diseases and/or pathology for detainees in police custody, and to determine how well those general health issues, diseases and/or pathology are being managed. This was done by a detailed analysis of healthcare issues of a cohort of detainees and reviewing intended and prescribed medication needs with current medication availability. In August 2007, a prospective detailed, anonymised, structured questionnaire survey was undertaken of 201 detainees in police custody in London, UK. Of these 83.6% consented to participate in the study. 85.1% of subjects were male; mean age was 33.9 years; 70.8% had English as a first language; 13.7% were of no fixed abode; 70.2% were registered with a general practitioner (primary care physician); 25% were already in contact with other healthcare teams; 7.1% had previously been sectioned under the Mental Health Act 1983; 16.7% had previously intentionally self-injured; 33.9% were dependent on heroin, 33.9% on crack cocaine; 25% on alcohol, 16.6% on benzodiazepines and 63.1% on cigarettes. 56% of subjects had active medical conditions; of those with active medical conditions 74% were prescribed medication for those medical conditions; only 3/70 had their medication available. 28/70 were not taking medication regularly, and many were not taking it at all. Three subjects who had deep vein thromboses were not taking their prescribed anticoagulants and six subjects with severe mental health issues were not taking their anti-psychotic medication. Mental health issues and depression predominated, but there was a very large range of mixed diseases and pathology. Asthma, epilepsy, diabetes, deep vein thrombosis, pulmonary embolism, hepatitis, and hypertension were all represented. The study has achieved its aims and has also shown that--in part because of the chaotic lifestyle of many detainees--appropriate care was not being rendered, thereby, putting both detainee, and potentially others coming into contact with them, at risk.


Region-specific onset of handling-induced changes in corticotropin-releasing factor and glucocorticoid receptor expression.
Early-life experience including maternal care profoundly influences hormonal stress responses during adulthood. Daily handling on postnatal day (P) 2-9, eliciting augmented maternal care upon returning pups to their cage, permanently modifies the expression of the stress neuromodulators corticotropin-releasing factor (CRF) and glucocorticoid receptor (GR). We have previously demonstrated reduced hypothalamic CRF expression already at the end of the handling period, followed by enhanced hippocampal GR mRNA levels (by P45). However, the initial site(s) and time of onset of these enduring changes have remained unclear. Therefore, we used semiquantitative in situ hybridization to delineate the spatiotemporal evolution of CRF and GR expression throughout stress-regulatory brain regions in handled (compared with undisturbed) pups. Enhanced CRF mRNA expression was apparent in the amygdaloid central nucleus (ACe) of handled pups already by P6. By P9, the augmented CRF mRNA levels persisted in ACe, accompanied by increased peptide expression in the bed nucleus of the stria terminalis and reduced expression in the paraventricular nucleus. The earliest change in GR consisted of reduced expression in the ACe of handled pups on P9, a time point when hippocampal GR expression was not yet affected. Thus, altered gene expression in ACe, bed nucleus of the stria terminalis as well as paraventricular nucleus may contribute to the molecular cascade by which handling (and increased maternal care) influences the stress response long term.


Computer-aided discovery of anti-HIV agents.
A review is provided on efforts in our laboratory over the last decade to discover anti-HIV agents. The work has focused on computer-aided design and synthesis of non-nucleoside inhibitors of HIV-1 reverse transcriptase (NNRTIs) with collaborative efforts on biological assaying and protein crystallography. Numerous design issues were successfully addressed including the need for potency against a wide range of viral variants, good aqueous solubility, and avoidance of electrophilic substructures. Computational methods including docking, de novo design, and free-energy perturbation (FEP) calculations made essential contributions. The result is novel NNRTIs with picomolar and low-nanomolar activities against wild-type HIV-1 and key variants that also show much improved solubility and lower cytotoxicity than recently approved drugs in the class.