A Notch-regulated proliferative stem cell zone in the developing spinal cord is an ancestral vertebrate trait.
Vertebrates have evolved the most sophisticated nervous systems we know. These differ from the nervous systems of invertebrates in several ways, including the evolution of new cell types, and the emergence and elaboration of patterning mechanisms to organise cells in time and space. Vertebrates also generally have many more cells in their central nervous systems than invertebrates, and an increase in neural cell number may have contributed to the sophisticated anatomy of the brain and spinal cord. Here, we study how increased cell number evolved in the vertebrate central nervous system, investigating the regulation of cell proliferation in the lamprey spinal cord. Markers of proliferation show that a ventricular progenitor zone is found throughout the lamprey spinal cord. We show that inhibition of Notch signalling disrupts the maintenance of this zone. When Notch is blocked, progenitor cells differentiate precociously, the proliferative ventricular zone is lost and differentiation markers become expressed throughout the spinal cord. Comparison with other chordates suggests that the emergence of a persistent Notch-regulated proliferative progenitor zone was a crucial step for the evolution of vertebrate spinal cord complexity.


The use of inhaled prostaglandins in patients with ARDS: a systematic review and meta-analysis.
This study aimed to determine whether inhaled prostaglandins are associated with improvement in pulmonary physiology or mortality in patients with ARDS and assess adverse effects. The following data sources were used: PubMed, EMBASE, CINAHL, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, reference lists, conference proceedings, and ClinicalTrials.gov. Studies selected included randomized controlled trials and nonrandomized studies. For data extraction, two reviewers independently screened titles and abstracts for eligibility. With regard to data synthesis, 25 studies (two RCTs) published over 21 years (1993-2014) were included. The PROSPERO registration number was CRD42014013180. One randomized controlled trial showed no difference in the change in mean Pao2 to Fio2 ratio when comparing inhaled alprostadil to placebo: 141.2 (95% CI, 120.8-161.5) to 161.5 (95% CI, 134.6-188.3) vs 163.4 (95% CI, 140.8-186.0) to 186.8 (95% CI, 162.9-210.7), P = .21. Meta-analysis of the remaining studies demonstrated that inhaled prostaglandins were associated with improvement in Pao2 to Fio2 ratio (16 studies; 39.0% higher; 95% CI, 26.7%-51.3%), and Pao2 (eight studies; 21.4% higher; 95% CI, 12.2%-30.6%), and a decrease in pulmonary artery pressure (-4.8 mm Hg; 95% CI, -6.8 mm Hg to -2.8 mm Hg). Risk of bias and heterogeneity were high. Meta-regression found no association with publication year (P = .862), baseline oxygenation (P = .106), and ARDS etiology (P = .816) with the treatment effect. Hypotension occurred in 17.4% of patients in observational studies. In ARDS, inhaled prostaglandins improve oxygenation and decrease pulmonary artery pressures and may be associated with harm. Data are limited both in terms of methodologic quality and demonstration of clinical benefit. The use of inhaled prostaglandins in ARDS needs further study.


A novel covalent post-synthetically modified MOF hybrid as a sensitive and selective fluorescent probe for Al3+ detection in aqueous media.
A modified MOF named UiO-66-NH2-SA was synthesized based on the covalent post synthetic attachment of the MOFs (UiO-66-NH2) and salicylaldehyde via a Schiff-base reaction. The as-prepared functionalized UiO-66-NH2-SA not only maintains its structural integrity during the PSM process, but also shows excellent luminescence and good fluorescence stability in water. It was further utilized as a novel fluorescent probe for detecting of Al3+. The fluorescence intensity of UiO-66-NH2-SA increased linearly upon increasing the concentration of Al3+ in the range of 0-500 μM with a detection limit of 6.98 μM. The possible mechanism is discussed. This study presents a new ratiometric and colorimetric Al3+ fluorescent sensor. The good fluorescence stability of UiO-66-NH2-SA in aqueous media, the low detection limit and the broad linear in sensing Al3+ indicate its high potential in practical applications.


Functional selectivity profiling of the angiotensin II type 1 receptor using pathway-wide BRET signaling sensors.
G protein-coupled receptors (GPCRs) are important therapeutic targets that exhibit functional selectivity (biased signaling), in which different ligands or receptor variants elicit distinct downstream signaling. Understanding all the signaling events and biases that contribute to both the beneficial and adverse effects of GPCR stimulation by given ligands is important for drug discovery. Here, we report the design, validation, and use of pathway-selective bioluminescence resonance energy transfer (BRET) biosensors that monitor the engagement and activation of signaling effectors downstream of G proteins, including protein kinase C (PKC), phospholipase C (PLC), p63RhoGEF, and Rho. Combined with G protein and β-arrestin BRET biosensors, our sensors enabled real-time monitoring of GPCR signaling at different levels in downstream pathways in both native and engineered cells. Profiling of the responses to 14 angiotensin II (AngII) type 1 receptor (AT1R) ligands enabled the clustering of compounds into different subfamilies of biased ligands and showed that, in addition to the previously reported functional selectivity between Gαq and β-arrestin, there are also biases among G protein subtypes. We also demonstrated that biases observed at the receptor and G protein levels propagated to downstream signaling pathways and that these biases could occur through the engagement of different G proteins to activate a common effector. We also used these tools to determine how naturally occurring AT1R variants affected signaling bias. This suite of BRET biosensors provides a useful resource for fingerprinting biased ligands and mutant receptors and for dissecting functional selectivity at various levels of GPCR signaling.


A voxel-based investigation for MRI-only radiotherapy of the brain using ultra short echo times.
Radiotherapy (RT) based on magnetic resonance imaging (MRI) as the only modality, so-called MRI-only RT, would remove the systematic registration error between MR and computed tomography (CT), and provide co-registered MRI for assessment of treatment response and adaptive RT. Electron densities, however, need to be assigned to the MRI images for dose calculation and patient setup based on digitally reconstructed radiographs (DRRs). Here, we investigate the geometric and dosimetric performance for a number of popular voxel-based methods to generate a so-called pseudo CT (pCT). Five patients receiving cranial irradiation, each containing a co-registered MRI and CT scan, were included. An ultra short echo time MRI sequence for bone visualization was used. Six methods were investigated for three popular types of voxel-based approaches; (1) threshold-based segmentation, (2) Bayesian segmentation and (3) statistical regression. Each approach contained two methods. Approach 1 used bulk density assignment of MRI voxels into air, soft tissue and bone based on logical masks and the transverse relaxation time T2 of the bone. Approach 2 used similar bulk density assignments with Bayesian statistics including or excluding additional spatial information. Approach 3 used a statistical regression correlating MRI voxels with their corresponding CT voxels. A similar photon and proton treatment plan was generated for a target positioned between the nasal cavity and the brainstem for all patients. The CT agreement with the pCT of each method was quantified and compared with the other methods geometrically and dosimetrically using both a number of reported metrics and introducing some novel metrics. The best geometrical agreement with CT was obtained with the statistical regression methods which performed significantly better than the threshold and Bayesian segmentation methods (excluding spatial information). All methods agreed significantly better with CT than a reference water MRI comparison. The mean dosimetric deviation for photons and protons compared to the CT was about 2% and highest in the gradient dose region of the brainstem. Both the threshold based method and the statistical regression methods showed the highest dosimetrical agreement.Generation of pCTs using statistical regression seems to be the most promising candidate for MRI-only RT of the brain. Further, the total amount of different tissues needs to be taken into account for dosimetric considerations regardless of their correct geometrical position.


Regulation of the ontogenic and regional expression of intestinal aminooligopeptidase.
Aminooligopeptidase (AOP) is the predominant peptidase in the small intestinal epithelium. During postnatal development in the rat, characteristic ontogenic and regional patterns of AOP activities become established. To investigate the molecular mechanisms regulating the maturational changes in AOP activity, we compared AOP synthesis and assembly in the jejunum of suckling (14-day-old) and weaned (28-day-old) rats. AOP synthesis was assessed in vivo by intraperitoneal labeling with [35S]methionine. Both AOP activity and AOP synthesis doubled at weaning, while its posttranslational processing remained unaltered. To examine the mechanisms responsible for generating the regional differences in AOP activity in weaned rats, we contrasted the de novo synthesis, kinetic properties, total immunoreactive protein, and degradation of the jejunal and ileal peptidases. Although AOP catalytic activity was significantly greater in the jejunum than in the ileum, its synthesis rate and substrate affinity (Km) were identical at both sites. However, the ileal peptidase was degraded more rapidly than the jejunal enzyme (t1/2 = 4 and 10 h, respectively). In summary, our data show that increased synthesis accounts for the ontogenic rise in intestinal AOP activity but that altered enzyme turnover produces the jejunal-ileal gradient in AOP activity in weaned rats. Thus the ontogenic and regional expressions of intestinal AOP are defined by transcriptional/translational and posttranscriptional regulatory mechanisms, respectively.


Working-memory capacity predicts the executive control of visual search among distractors: the influences of sustained and selective attention.
Variation in working-memory capacity (WMC) predicts individual differences in only some attention-control capabilities. Whereas higher WMC subjects outperform lower WMC subjects in tasks requiring the restraint of prepotent but inappropriate responses, and the constraint of attentional focus to target stimuli against distractors, they do not differ in prototypical visual-search tasks, even those that yield steep search slopes and engender top-down control. The present three experiments tested whether WMC, as measured by complex memory span tasks, would predict search latencies when the 1-8 target locations to be searched appeared alone, versus appearing among distractor locations to be ignored, with the latter requiring selective attentional focus. Subjects viewed target-location cues and then fixated on those locations over either long (1,500-1,550 ms) or short (300 ms) delays. Higher WMC subjects identified targets faster than did lower WMC subjects only in the presence of distractors and only over long fixation delays. WMC thus appears to affect subjects' ability to maintain a constrained attentional focus over time.


Polarized beam splitting effect in inhomogeneously magnetized magnetooptic films.
Linearly polarized light passing through a several micron thick magnetooptic film in the inhomogeneous magnetization state is split into a linearly polarized central beam and linearly polarized first and higher order diverging rings. The polarization of the central output beam lies in the same direction as the linearly polarized input, while the polarization of the diverging rings lies in a direction orthogonal to the input plane of polarization. The effect is described, and applications of the effect are discussed.


Identification of human sympathetic neurovascular control using multivariate wavelet decomposition analysis.
The dynamic regulation of cerebral blood flow (CBF) is thought to involve myogenic and chemoreflex mechanisms, but the extent to which the sympathetic nervous system also plays a role remains debated. Here we sought to identify the role of human sympathetic neurovascular control by examining cerebral pressure-flow relations using linear transfer function analysis and multivariate wavelet decomposition analysis that explicitly accounts for the confounding effects of dynamic end-tidal Pco2 (PetCO2 ) fluctuations. In 18 healthy participants randomly assigned to the α1-adrenergic blockade group (n = 9; oral Prazosin, 0.05 mg/kg) or the placebo group (n = 9), we recorded blood pressure, middle cerebral blood flow velocity, and breath-to-breath PetCO2 Analyses showed that the placebo administration did not alter wavelet phase synchronization index (PSI) values, whereas sympathetic blockade increased PSI for frequency components ≤0.03 Hz. Additionally, three-way interaction effects were found for PSI change scores, indicating that the treatment response varied as a function of frequency and whether PSI values were PetCO2 corrected. In contrast, sympathetic blockade did not affect any linear transfer function parameters. These data show that very-low-frequency CBF dynamics have a composite origin involving, not only nonlinear and nonstationary interactions between BP and PetCO2 , but also frequency-dependent interplay with the sympathetic nervous system.


Visual neglect associated with frontal lobe infarction.
Five patients with left-sided visual neglect following focal infarction of the right frontal lobe are presented. Lesion location was assessed using computed tomography or magnetic resonance imaging. The common area of lesion overlap was small, being confined to the dorsal aspect of the inferior frontal gyrus (Brodmann's area 44) and the immediate underlying white matter. This cortical region is part of the homologue of Broca's area in the right hemisphere and is considered to be part of human premotor cortex. The association of neglect with injury to this area suggests it may play an important role in directing attention in visual space.


Membrane particles from Escherichia coli and Bacillus subtilis, containing penicillin-binding proteins and enriched for chromosomal-origin DNA.
Rapid-sedimenting DNA-membrane complexes were obtained from both Bacillus subtilis and Escherichia coli by a method involving gentle lysis followed by restriction enzyme digestion and sucrose gradient fractionation. These complexes were substantially enriched in chromosomal origin DNA, and in B. subtilis, the complexes were enriched in penicillin-binding proteins relative to that of the total membrane. Such complexes may represent procaryotic membrane domains which are topographically and functionally distinct.


The intensification of the water footprint of hydraulic fracturing.
Unconventional oil and gas exploration in the United States has experienced a period of rapid growth, followed by several years of limited production due to falling and low natural gas and oil prices. Throughout this transition, the water use for hydraulic fracturing and wastewater production in major shale gas and oil production regions has increased; from 2011 to 2016, the water use per well increased up to 770%, while flowback and produced water volumes generated within the first year of production increased up to 1440%. The water-use intensity (that is, normalized to the energy production) increased ubiquitously in all U.S. shale basins during this transition period. The steady increase of the water footprint of hydraulic fracturing with time implies that future unconventional oil and gas operations will require larger volumes of water for hydraulic fracturing, which will result in larger produced oil and gas wastewater volumes.


Defective insulin secretion by chronic glucagon receptor activation in glucose intolerant mice.
Stimulation of insulin secretion by short-term glucagon receptor (GCGR) activation is well characterized; however, the effect of long-term GCGR activation on β-cell function is not known, but of interest, since hyperglucagonemia occurs early during development of type 2 diabetes. Therefore, we examined whether chronic GCGR activation affects insulin secretion in glucose intolerant mice. To induce chronic GCGR activation, high-fat diet fed mice were continuously (2 weeks) infused with the stable glucagon analog ZP-GA-1 and challenged with oral glucose and intravenous glucose±glucagon-like peptide 1 (GLP1). Islets were isolated to evaluate the insulin secretory response to glucose±GLP1 and their pancreas were collected for immunohistochemical analysis. Two weeks of ZP-GA-1 infusion reduced insulin secretion both after oral and intravenous glucose challenges in vivo and in isolated islets. These inhibitory effects were corrected for by GLP1. Also, we observed increased β-cell area and islet size. We conclude that induction of chronic ZP-GA-1 levels in glucose intolerant mice markedly reduces insulin secretion, and thus, we suggest that chronic activation of the GCGR may contribute to the failure of β-cell function during development of type 2 diabetes.


Prospective study of zirconia-based restorations: 3-year clinical results.
To evaluate fixed dental prostheses (FDPs) and single crowns made of zirconia substructures veneered with new fluorapatite veneering porcelain. Thirty-eight patients received 18 FDPs and 50 single crowns. Zirconia substructures were fabricated by a CAD/CAM system and veneered using the powder buildup technique. All restorations were cemented with glass ionomer. Baseline evaluation was performed 2 weeks after cementation with recall examinations at 12, 24, and 36 months by calibrated investigators. SEM was performed on replicas of all restorations. Survival probabilities according to Kaplan-Meier were calculated. The mean service time was 35 (+/- 14) months. After 3 years of clinical service, three biologic and five technical failures were recorded. All failures occurred in the FDP group. One FDP was removed after biologic failure of one abutment tooth. The Kaplan-Meier survival probability was 88.2% after 35 months for all types of failures and 98.5% concerning restorations in service. The type of restoration showed significant influence on the survival probability (P < .001, log-rank test). No influence on gingival parameters from the restorations was observed compared to control teeth. Zirconia substructures veneered with fluorapatite veneering porcelain seem to be a reliable treatment option. More research has to be conducted to investigate differences between single crowns and FDPs.


Anodal transcranial direct current stimulation over the primary motor cortex does not enhance the learning benefits of self-controlled feedback schedules.
A distinct learning advantage has been shown when participants control their knowledge of results (KR) scheduling during practice compared to when the same KR schedule is imposed on the learner without choice (i.e., yoked schedules). Although the learning advantages of self-controlled KR schedules are well-documented, the brain regions contributing to these advantages remain unknown. Identifying key brain regions would not only advance our theoretical understanding of the mechanisms underlying self-controlled learning advantages, but would also highlight regions that could be targeted in more applied settings to boost the already beneficial effects of self-controlled KR schedules. Here, we investigated whether applying anodal transcranial direct current stimulation (tDCS) to the primary motor cortex (M1) would enhance the typically found benefits of learning a novel motor skill with a self-controlled KR schedule. Participants practiced a spatiotemporal task in one of four groups using a factorial combination of KR schedule (self-controlled vs. yoked) and tDCS (anodal vs. sham). Testing occurred on two consecutive days with spatial and temporal accuracy measured on both days and learning was assessed using 24-h retention and transfer tests without KR. All groups improved their performance in practice and a significant effect for practicing with a self-controlled KR schedule compared to a yoked schedule was found for temporal accuracy in transfer, but a similar advantage was not evident in retention. There were no significant differences as a function of KR schedule or tDCS for spatial accuracy in retention or transfer. The lack of a significant tDCS effect suggests that M1 may not strongly contribute to self-controlled KR learning advantages; however, caution is advised with this interpretation as typical self-controlled learning benefits were not strongly replicated in the present experiment.


Broadband third harmonic generation in tapered silica fibres.
Optical microfibres have recently attracted much attention for nonlinear applications, due to their tight modal confinement. Here, we report broadband third harmonic generation based on the intermodal phase matching technique in silica microfibres of several centimetres. The third harmonic signal is predominantly generated from the taper transition regions (rather than the waist), wherein the range of diameters permits phase matching over a wide bandwidth. Microfibres up to 4.5 cm long were fabricated with waist diameters below 2.5 μm to allow a λ = 1.55 μm pump to phase match with several higher order third harmonic modes; conversion rates up to 3 × 10⁻⁴ were recorded when pumped with 4 ns pulses at a peak power of 1.25 kW. Analysis of the third harmonic frequencies generated from the nonlinearly broadened pump components indicate a 5 dB conversion bandwidth of at least 36 nm, with harmonic power detected over a 150 nm range.


Woodlot management and livelihoods in a tropical conservation landscape.
In biodiversity hotspots, there is often tension between human needs and conservation, exacerbated when protected areas prevent access to natural resources. Forest-dependent people may compensate for exclusion by managing unprotected forests or cultivating planted woodlots. Outside Bwindi Impenetrable National Park in Uganda, household wood product needs are high and population growth puts pressure on the environment. We investigated the role of privately and collectively managed woodlots in provisioning wood products and supporting local livelihoods. We found that households relied heavily on woodlots for daily needs and as resources during time of need. We also found that locally relevant social institutions, called stretcher groups, played a role in the management of woodlots, providing shared community resources. Privately and collectively owned woodlots support local livelihoods and wood product needs in the region. Long-term management of forests in Uganda should consider the value of woodlots and the mechanisms required to support them.


Role of disulfide bonds in the structure and potassium channel blocking activity of ShK toxin.
ShK toxin, a potassium channel blocker from the sea anemone Stichodactyla helianthus, is a 35 residue polypeptide cross-linked by three disulfide bridges: Cys3-Cys35, Cys12-Cys28, and Cys17-Cys32. To investigate the role of these disulfides in the structure and channel-blocking activity of ShK toxin, a series of analogues was synthesized by selective replacement of each pair of half-cystines with two alpha-amino-butyrate (Abu) residues. The remaining two disulfide pairs were formed unambiguously using an orthogonal protecting group strategy of Cys(Trt) or Cys(Acm) at the appropriate position. The peptides were tested in vitro for their ability to block Kv1.1 and Kv1.3 potassium channels and their ability to displace [(125)I]dendrotoxin binding to rat brain synaptosomal membranes. The monocyclic peptides showed no activity in these assays. Of the dicyclic peptides, [Abu12,28]ShK(3-35,17)(-)(32) (where the subscript indicates disulfide connectivities) had weak activity on Kv1.3 and Kv1.1. [Abu17,32]ShK(3-35,12)(-)(28) blocked Kv1.3 with low nanomolar potency, but was less effective (being comparable to [Abu12,28]ShK(3-35,17)(-)(32)) against Kv1.1. [Abu3, 35]ShK(12-28,17)(-)(32), retained high picomolar affinity against both channels. Corroborating these results, [Abu3,35]ShK(12-28, 17)(-)(32) had an IC(50) ratio relative to native toxin of 18 in the displacement assay, whereas [Abu17,32]ShK(3-35,12)(-)(28) and [Abu12, 28]ShK(3-35,17)(-)(32) had ratios of 69 and 390, respectively. Thus, the disulfide bond linking the N- and C-terminal regions is less important for activity than the internal disulfides. NMR analysis of the [Abu12,28] and [Abu17,32] analogues indicated that they had little residual structure, consistent with their significantly reduced activities. By contrast, [Abu3,35]ShK(12-28,17)(-)(32) had a moderately well-defined solution structure, with a mean pairwise root-mean-square deviation of 1.33 A over the backbone heavy atoms. This structure nevertheless showed significant differences from that of native ShK toxin. The possible interactions of this analogue with the channel and the distinction between native secondary and tertiary structure on one hand and global topology imposed by the disulfide bridges on the other are discussed.


Construction of vectors for inducible and constitutive gene expression in Lactobacillus.
Microarray analysis of the genome of Lactobacillus acidophilus identified a number of operons that were differentially expressed in response to carbohydrate source or constitutively expressed regardless of carbohydrate source. These included operons implicated in the transport and catabolism of fructooligosaccharides (FOS), lactose (lac), trehalose (tre) and genes directing glycolysis. Analysis of these operons identified a number of putative promoter and repressor elements, which were used to construct a series of expression vectors for use in lactobacilli, based on the broad host range pWV01 replicon. A β-glucuronidase (GusA3) reporter gene was cloned into each vector to characterize expression from each promoter. GUS reporter assays showed FOS, lac and tre based vectors to be highly inducible by their specific carbohydrate and repressed by glucose. Additionally, a construct based on the phosphoglycerate mutase (pgm) promoter was constitutively highly expressed. To demonstrate the potential utility of these vectors, we constructed a plasmid for the overexpression of the oxalate degradation pathway (Frc and Oxc) of L. acidophilus NCFM. This construct was able to improve oxalate degradation by L. gasseri ATCC 33323 and compliment a L. acidophilus oxalate-deficient mutant. Development of these expression vectors could support several novel applications, including the expression of enzymes, proteins, vaccines and biotherapeutics by intestinal lactobacilli.


Structure of pvu II DNA-(cytosine N4) methyltransferase, an example of domain permutation and protein fold assignment.
We have determined the structure of Pvu II methyltransferase (M. Pvu II) complexed with S -adenosyl-L-methionine (AdoMet) by multiwavelength anomalous diffraction, using a crystal of the selenomethionine-substituted protein. M. Pvu II catalyzes transfer of the methyl group from AdoMet to the exocyclic amino (N4) nitrogen of the central cytosine in its recognition sequence 5'-CAGCTG-3'. The protein is dominated by an open alpha/beta-sheet structure with a prominent V-shaped cleft: AdoMet and catalytic amino acids are located at the bottom of this cleft. The size and the basic nature of the cleft are consistent with duplex DNA binding. The target (methylatable) cytosine, if flipped out of the double helical DNA as seen for DNA methyltransferases that generate 5-methylcytosine, would fit into the concave active site next to the AdoMet. This M. Pvu IIalpha/beta-sheet structure is very similar to those of M. Hha I (a cytosine C5 methyltransferase) and M. Taq I (an adenine N6 methyltransferase), consistent with a model predicting that DNA methyltransferases share a common structural fold while having the major functional regions permuted into three distinct linear orders. The main feature of the common fold is a seven-stranded beta-sheet (6 7 5 4 1 2 3) formed by five parallel beta-strands and an antiparallel beta-hairpin. The beta-sheet is flanked by six parallel alpha-helices, three on each side. The AdoMet binding site is located at the C-terminal ends of strands beta1 and beta2 and the active site is at the C-terminal ends of strands beta4 and beta5 and the N-terminal end of strand beta7. The AdoMet-protein interactions are almost identical among M. Pvu II, M. Hha I and M. Taq I, as well as in an RNA methyltransferase and at least one small molecule methyltransferase. The structural similarity among the active sites of M. Pvu II, M. Taq I and M. Hha I reveals that catalytic amino acids essential for cytosine N4 and adenine N6 methylation coincide spatially with those for cytosine C5 methylation, suggesting a mechanism for amino methylation.


Teledermatology for diagnosing skin cancer in adults.
Early accurate detection of all skin cancer types is essential to guide appropriate management and to improve morbidity and survival. Melanoma and squamous cell carcinoma (SCC) are high-risk skin cancers which have the potential to metastasise and ultimately lead to death, whereas basal cell carcinoma (BCC) is usually localised with potential to infiltrate and damage surrounding tissue. Anxiety around missing early curable cases needs to be balanced against inappropriate referral and unnecessary excision of benign lesions. Teledermatology provides a way for generalist clinicians to access the opinion of a specialist dermatologist for skin lesions that they consider to be suspicious without referring the patients through the normal referral pathway. Teledermatology consultations can be 'store-and-forward' with electronic digital images of a lesion sent to a dermatologist for review at a later time, or can be live and interactive consultations using videoconferencing to connect the patient, referrer and dermatologist in real time. To determine the diagnostic accuracy of teledermatology for the detection of any skin cancer (melanoma, BCC or cutaneous squamous cell carcinoma (cSCC)) in adults, and to compare its accuracy with that of in-person diagnosis. We undertook a comprehensive search of the following databases from inception up to August 2016: Cochrane Central Register of Controlled Trials, MEDLINE, Embase, CINAHL, CPCI, Zetoc, Science Citation Index, US National Institutes of Health Ongoing Trials Register, NIHR Clinical Research Network Portfolio Database and the World Health Organization International Clinical Trials Registry Platform. We studied reference lists and published systematic review articles. Studies evaluating skin cancer diagnosis for teledermatology alone, or in comparison with face-to-face diagnosis by a specialist clinician, compared with a reference standard of histological confirmation or clinical follow-up and expert opinion. We also included studies evaluating the referral accuracy of teledermatology compared with a reference standard of face-to-face diagnosis by a specialist clinician. Two review authors independently extracted all data using a standardised data extraction and quality assessment form (based on QUADAS-2). We contacted authors of included studies where there were information related to the target condition of any skin cancer missing. Data permitting, we estimated summary sensitivities and specificities using the bivariate hierarchical model. Due to the scarcity of data, we undertook no covariate investigations for this review. For illustrative purposes, we plotted estimates of sensitivity and specificity on coupled forest plots for diagnostic threshold and target condition under consideration. The review included 22 studies reporting diagnostic accuracy data for 4057 lesions and 879 malignant cases (16 studies) and referral accuracy data for reported data for 1449 lesions and 270 'positive' cases as determined by the reference standard face-to-face decision (six studies). Methodological quality was variable with poor reporting hindering assessment. The overall risk of bias was high or unclear for participant selection, reference standard, and participant flow and timing in at least half of all studies; the majority were at low risk of bias for the index test. The applicability of study findings were of high or unclear concern for most studies in all domains assessed due to the recruitment of participants from secondary care settings or specialist clinics rather than from primary or community-based settings in which teledermatology is more likely to be used and due to the acquisition of lesion images by dermatologists or in specialist imaging units rather than by primary care clinicians.Seven studies provided data for the primary target condition of any skin cancer (1588 lesions and 638 malignancies). For the correct diagnosis of lesions as malignant using photographic images, summary sensitivity was 94.9% (95% confidence interval (CI) 90.1% to 97.4%) and summary specificity was 84.3% (95% CI 48.5% to 96.8%) (from four studies). Individual study estimates using dermoscopic images or a combination of photographic and dermoscopic images generally suggested similarly high sensitivities with highly variable specificities. Limited comparative data suggested similar diagnostic accuracy between teledermatology assessment and in-person diagnosis by a dermatologist; however, data were too scarce to draw firm conclusions. For the detection of invasive melanoma or atypical intraepidermal melanocytic variants both sensitivities and specificities were more variable. Sensitivities ranged from 59% (95% CI 42% to 74%) to 100% (95% CI 48% to 100%) and specificities from 30% (95% CI 22% to 40%) to 100% (95% CI 93% to 100%), with reported diagnostic thresholds including the correct diagnosis of melanoma, classification of lesions as 'atypical' or 'typical, and the decision to refer or to excise a lesion.Referral accuracy data comparing teledermatology against a face-to-face reference standard suggested good agreement for lesions considered to require some positive action by face-to-face assessment (sensitivities of over 90%). For lesions considered of less concern when assessed face-to-face (e.g. for lesions not recommended for excision or referral), agreement was more variable with teledermatology specificities ranging from 57% (95% CI 39% to 73%) to 100% (95% CI 86% to 100%), suggesting that remote assessment is more likely recommend excision, referral or follow-up compared to in-person decisions. Studies were generally small and heterogeneous and methodological quality was difficult to judge due to poor reporting. Bearing in mind concerns regarding the applicability of study participants and of lesion image acquisition in specialist settings, our results suggest that teledermatology can correctly identify the majority of malignant lesions. Using a more widely defined threshold to identify 'possibly' malignant cases or lesions that should be considered for excision is likely to appropriately triage those lesions requiring face-to-face assessment by a specialist. Despite the increasing use of teledermatology on an international level, the evidence base to support its ability to accurately diagnose lesions and to triage lesions from primary to secondary care is lacking and further prospective and pragmatic evaluation is needed.


Human stem cell delivery for treatment of large segmental bone defects.
Local or systemic stem cell delivery has the potential to promote repair of a variety of damaged or degenerated tissues. Although various stem cell sources have been investigated for bone repair, few comparative reports exist, and cellular distribution and viability postimplantation remain key issues. In this study, we quantified the ability of tissue-engineered constructs containing either human fetal or adult stem cells to enhance functional repair of nude rat critically sized femoral defects. After 12 weeks, defects treated with cell-seeded polymer scaffolds had significantly higher bone ingrowth and torsional strength compared to those receiving acellular scaffolds, although there were no significant differences between the cell sources. Next, stem cells were labeled with fluorescent quantum dots (QDs) in an attempt to noninvasively track their distribution after delivery on scaffolds. Clear fluorescence was observed at implantation sites throughout the study; however, beginning 7-10 days after surgery, signals were also observed at contralateral sites treated with acellular QD-free scaffolds. Although immunostaining for human nuclei revealed retention of some cells at the implantation site, no human cells were detected in the control limb defects. Additional histological analysis of implantation and control defect tissues revealed macrophages containing endocytosed QDs. Furthermore, QD-labeling appeared to diminish transplanted cell function resulting in reduced healing responses. In summary, augmentation of polymeric scaffolds with stem cells derived from fetal and adult tissues significantly enhanced healing of large segmental bone defects; however, QD labeling of stem cells eliminated the observed therapeutic effect and failed to conclusively track stem cell location long-term in vivo.


Role of granulocytes in the prevention and therapy of experimental Streptococcus sanguis endocarditis in rabbits.
The contributions of granulocytes to the prevention and therapy of Streptococcus sanguis endocarditis with procaine benzylpenicillin (PBP) was investigated in rabbits. Depletion of granulocytes by treatment with mechlorethamine appeared to have no significant effect on either the prophylactic or therapeutic activities of PBP. Administration of 3,000 IU of PBP before inoculation with S. sanguis retarded the course of the endocarditis for only 24 h whether granulocytes were normal or depressed in numbers. Prophylaxis with either 15,000 or 30,000 IU of PBP was equally effective in non-granulocytopenic and granulocytopenic rabbits. Treatment of established infections with PBP at doses of 3,000 to 300,000 IU of PBP at 12-h intervals for 48 h was equally effective in rabbits with normal and depressed numbers of granulocytes. The effect of 3,000 IU of PBP was equivalent, however, to that of granulocytes alone, as shown by the fact that the numbers of CFU per gram of vegetation in the granulocytopenic rabbits treated with this dose of PBP and in the non-PBP-treated control rabbits were not significantly different.


The adaptation of a Danish version of the Pain Self-Efficacy Questionnaire: reliability and construct validity in a population of patients with fibromyalgia in Denmark.
The aim of this study was to translate, culturally adapt and evaluate the psychometric properties of the Pain Self-Efficacy Questionnaire (PSEQ) in a population of patients with fibromyalgia in Denmark. The study sample included 102 patients diagnosed with fibromyalgia referred to a specialist clinic. The PSEQ was translated and adapted to a Danish setting using a standard stepwise forward-backward translation procedure, followed by initial testing and focus group interview. Reliability was examined by analysing internal consistency and test-retest agreement. Construct validity was examined by investigating dimensionality, targeting, local independence, category functioning and differential item functioning (DIF). Reliability was high: Cronbach's alpha 0.88, test-retest correlation 0.93, intraclass correlation coefficient (ICC) 0.89 and item-total correlations 0.44-0.70. Factor analyses and item response (IRT) models indicated unidimensionality, and the PSEQ-DK was well targeted to the sample. High interitem correlation was observed between two items, indicating local dependence, and item misfit and DIF were observed for a few items. However, the overall fit of the scale to a single-factor model and IRT models supported acceptable construct validity. The PSEQ-DK showed acceptable psychometric properties and can therefore represent a reliable and valid measure for evaluating self-efficacy in patients with fibromyalgia in Denmark.


3 versus 6 months of adjuvant oxaliplatin-fluoropyrimidine combination therapy for colorectal cancer (SCOT): an international, randomised, phase 3, non-inferiority trial.
6 months of oxaliplatin-containing chemotherapy is usually given as adjuvant treatment for stage 3 colorectal cancer. We investigated whether 3 months of oxaliplatin-containing chemotherapy would be non-inferior to the usual 6 months of treatment. The SCOT study was an international, randomised, phase 3, non-inferiority trial done at 244 centres. Patients aged 18 years or older with high-risk stage II and stage III colorectal cancer underwent central randomisation with minimisation for centre, choice of regimen, sex, disease site, N stage, T stage, and the starting dose of capecitabine. Patients were assigned (1:1) to receive 3 months or 6 months of adjuvant oxaliplatin-containing chemotherapy. The chemotherapy regimens could consist of CAPOX (capecitabine and oxaliplatin) or FOLFOX (bolus and infused fluorouracil with oxaliplatin). The regimen was selected before randomisation in accordance with choices of the patient and treating physician. The primary study endpoint was disease-free survival and the non-inferiority margin was a hazard ratio of 1·13. The primary analysis was done in the intention-to-treat population and safety was assessed in patients who started study treatment. This trial is registered with ISRCTN, number ISRCTN59757862, and follow-up is continuing. 6088 patients underwent randomisation between March 27, 2008, and Nov 29, 2013. The intended treatment was FOLFOX in 1981 patients and CAPOX in 4107 patients. 3044 patients were assigned to 3 month group and 3044 were assigned to 6 month group. Nine patients in the 3 month group and 14 patients in the 6 month group did not consent for their data to be used, leaving 3035 patients in the 3 month group and 3030 patients in the 6 month group for the intention-to-treat analyses. At the cutoff date for analysis, there had been 1482 disease-free survival events, with 740 in the 3 month group and 742 in the 6 month group. 3 year disease-free survival was 76·7% (95% CI 75·1-78·2) for the 3 month group and 77·1% (75·6-78·6) for the 6 month group, giving a hazard ratio of 1·006 (0·909-1·114, test for non-inferiority p=0·012), significantly below the non-inferiority margin. Peripheral neuropathy of grade 2 or worse was more common in the 6 month group (237 [58%] of 409 patients for the subset with safety data) than in the 3 month group (103 [25%] of 420) and was long-lasting and associated with worse quality of life. 1098 serious adverse events were reported (492 reports in the 3 month group and 606 reports in the 6 month group) and 32 treatment-related deaths occurred (16 in each group). In the whole study population, 3 months of oxaliplatin-containing adjuvant chemotherapy was non-inferior to 6 months of the same therapy for patients with high-risk stage II and stage III colorectal cancer and was associated with reduced toxicity and improved quality of life. Despite the fact the study was underpowered, these data suggest that a shorter duration leads to similar survival outcomes with better quality of life and thus might represent a new standard of care. Medical Research Council, Swedish Cancer Society, NETSCC, and Cancer Research UK.


Scaled deployment of Wolbachia to protect the community from dengue and other Aedes transmitted arboviruses.
Background: A number of new technologies are under development for the control of mosquito transmitted viruses, such as dengue, chikungunya and Zika that all require the release of modified mosquitoes into the environment. None of these technologies has been able to demonstrate evidence that they can be implemented at a scale beyond small pilots. Here we report the first successful citywide scaled deployment of Wolbachia in the northern Australian city of Townsville. Methods: The wMel strain of Wolbachia was backcrossed into a local Aedes aegypti genotype and mass reared mosquitoes were deployed as eggs using mosquito release containers (MRCs). In initial stages these releases were undertaken by program staff but in later stages this was replaced by direct community release including the development of a school program that saw children undertake releases. Mosquito monitoring was undertaken with Biogents Sentinel (BGS) traps and individual mosquitoes were screened for the presence of Wolbachia with a Taqman qPCR or LAMP diagnostic assay. Dengue case notifications from Queensland Health Communicable Disease Branch were used to track dengue cases in the city before and after release. Results: Wolbachia was successfully established into local Ae. aegypti mosquitoes across 66 km 2 in four stages over 28 months with full community support. A feature of the program was the development of a scaled approach to community engagement. Wolbachia frequencies have remained stable since deployment and to date no local dengue transmission has been confirmed in any area of Townsville after Wolbachia has established, despite local transmission events every year for the prior 13 years and an epidemiological context of increasing imported cases. Conclusion: Deployment of Wolbachia into Ae. aegypti populations can be readily scaled to areas of ~60km 2 quickly and cost effectively and appears in this context to be effective at stopping local dengue transmission.


Bisphosphonates for osteoporosis in people with cystic fibrosis.
Osteoporosis is a disorder of bone mineralization that can lead to reduced bone mineral density and an increased risk for fractures. It is found in about one third of adults with cystic fibrosis. Bisphosphonates have been shown to increase bone mineral density and decrease the risk of new fractures in post-menopausal women and in patients receiving long-term oral corticosteroids. To assess the effects of bisphosphonates on the frequency of fractures, bone mineral density, quality of life, adverse events, study withdrawals, and survival in people with cystic fibrosis. Relevant trials were identified in the Cochrane Cystic Fibrosis and Genetic Disorders Review Group specialised register of controlled trials by the centre co-ordinator. This register comprises references identified from comprehensive electronic database searches, handsearching of relevant journals and of conference proceedings. Additional sources such as abstract books for osteoporosis conferences were hand-searched by the authors. The most recent search(es) were conducted in February 2001. Randomised controlled trials of at least six months duration that studied the use of bisphosphonates in adults with cystic fibrosis were considered for inclusion. Outcomes included one of the following: fractures, bone mineral density, quality of life, adverse events, study withdrawals, or survival. Information on study design, participants, interventions, and outcomes was abstracted from included studies. Two independent reviewers abstracted the information. Authors were contacted to obtain missing data. Two trials were identified in the trials search. Both trials with a total of 65 participants were included in this review. One study examined patients without lung transplants while the other study included only patients who had received a lung transplant. The intervention in both trials was pamidronate administered intravenously every three months. In patients who had not received a lung transplant, bone mineral density at axial sites was increased after six months of treatment in the treatment group compared to the control group (lumbar spine WMD [for % change BMD] = 5.8 [95% CI 4.63 to 6.97], hip WMD = 3.00 [95% CI 1.99 to 4.01]). There was a small decrease in forearm bone mineral density in patients treated with pamidronate versus controls (distal forearm WMD = -1.70 [95% CI -2.46 to -0.94]). Bone pain was the most common adverse event occurring in 11/15 participants not using corticosteroids (RR = 24.40, 95% CI 1.57 to 381.48). There was no difference in survival (RR = 1.00, 95% CI 0.83 to 1.20), although this may be due to short follow-up and small sample size. In patients who had received a lung transplant, the number of new fractures did not change with the use of pamidronate (non-vertebral RR = 3.38 [95% CI 0.39 to 29.29], vertebral RR = 0.56 [95% CI 0.17 to 1.89]). Bone mineral density at axial sites was increased after two years of treatment in the treatment group compared to the control group (lumbar spine WMD [for % change in BMD] = 6.20 [95% CI 4.28 to 8.12], femur WMD = 7.90 [95% CI 5.78 to 10.03]). Intravenous pamidronate increases bone mineral density at axial sites in people with cystic fibrosis, although it can cause severe bone pain in patients not receiving corticosteroids. Additional studies in larger populations are needed to determine the effect on fracture rate and survival.


Detection of S83V GyrA mutation in quinolone-resistant Shewanella algae using comparative genomics.
Shewanella algae is a zoonotic pathogen that poses a serious health threat to immunocompromised hosts. Treatment of S. algae infections is challenging due to the pathogen's intrinsic resistance to a variety of β-lactam antibiotics. Therapeutic options have become further limited by the emergence of quinolone-resistant strains. Currently, there are few studies concerning the genetic and molecular mechanisms underlying acquired quinolones resistance in S. algae. qnrA was once proposed as the candidate gene related to quinolones resistance in S. algae. However, recent studies demonstrated qnrA are highly conservative and does not confer resistance to quinolones in S. algae. A total of 27 non-duplicated isolates of S. algae strains were examined. MICs of ciprofloxacin were determined using Vitek 2. Whole genome sequencing was performed using MiSeq platform. Comprehensive Antibiotic Resistance Database and ResFinder were used for annotation of quinolones resistance genes. Multiple sequence alignment by EMBOSS Clustal Omega were used to identified mutation in quinolone resistance-determining regions. To investigation of the alteration of protein structure induced by mutation, in silico molecular docking studies was conducted using Accryl Discovery studio visualizer. All S. algae harbored the quinolone-resistance associated genes (qnrA, gyrA, gyrB, parC, and parE) regardless its resistance to ciprofloxacin. Comparison of these genomes identified a nonsynonymous mutation (S83V) in chromosome-encoded gyrase subunits (GyrA) in quinolone-resistant strain. We found this mutation disrupts the water-metal ion bridge, reduces the affinity of the quinolone-enzyme complex for the metal ions and therefore decrease the capability of quinolones to stabilize cleavage complexes. The study provides insight into the quinolone resistance mechanisms in S. algae, which would be helpful for the evolution of antibiotic resistance in this bacterium.


Contemporary protease inhibitors and cardiovascular risk.
To review the evidence linking use of HIV protease inhibitors with excess risk of cardiovascular disease (CVD) in HIV+ populations. For the two contemporary most frequently used protease inhibitors, darunavir and atazanavir [both pharmacologically boosted with ritonavir (/r)], darunavir/r has been shown to be associated with increased CVD risk. The effect is cumulative with longer exposure increasing risk and an effect size comparable to what has been observed for previously developed protease inhibitors. Biological mechanisms may be overlapping and include perturbed lipid metabolism and accumulation of cholesterol derivatives within macrophages. Conversely, atazanavir/r has not been shown to be associated with CVD, possibly because of its ability to increase cardioprotective bilirubin levels. Evidence linking protease inhibitors to CVD is based on observational studies only, whereas plausible biological explanations are well established and derived from randomized trials and controlled experiments. Given the possible association with clinical disease, a conservative approach to apply the data in daily practise is proposed which is focused on individualization of care based on underlying risk of CVD.


COBRA encodes a putative GPI-anchored protein, which is polarly localized and necessary for oriented cell expansion in Arabidopsis.
To control organ shape, plant cells expand differentially. The organization of the cellulose microfibrils in the cell wall is a key determinant of differential expansion. Mutations in the COBRA (COB) gene of Arabidopsis, known to affect the orientation of cell expansion in the root, are reported here to reduce the amount of crystalline cellulose in cell walls in the root growth zone. The COB gene, identified by map-based cloning, contains a sequence motif found in proteins that are anchored to the extracellular surface of the plasma membrane through a glycosylphosphatidylinositol (GPI) linkage. In animal cells, this lipid linkage is known to confer polar localization to proteins. The COB protein was detected predominately on the longitudinal sides of root cells in the zone of rapid elongation. Moreover, COB RNA levels are dramatically upregulated in cells entering the zone of rapid elongation. Based on these results, models are proposed for the role of COB as a regulator of oriented cell expansion.


Gastrointestinal myoelectric correlates of vomiting in the dog.
Prior to vomiting, two gastrointestinal contractile events occur in succession: a giant contraction that propagates retrograde from mid-small intestine to the antrum (RGC) and a series of phasic contractions that occur at all levels of the gastrointestinal tract. We quantitatively examined the gastrointestinal myoelectric events associated with vomiting and correlated them with the previously defined contractile events. Twelve dogs of either sex were implanted with electrical or contractile recording devices implanted on the stomach and small intestine. After an overnight fast, gut activity was recorded before and after apomorphine administration (2.5-15 micrograms/kg, iv). We found that the RGC was correlated with two successive electrical events: disruption of electrical control activity (ECA) cycling and a potential change occurring at the upstroke of the RGC. However, the initial myoelectric event associated with vomiting was ECA frequency slowing of the antrum and lower half of the small intestine. The post-RGC phasic contractions were associated with ECA-dependent response activity and occurred during the period of ECA frequency slowing. Atropine (100 micrograms/kg iv) blocked the RGC and its associated electrical events but not the slowing of ECA frequency. Supradiaphragmatic vagotomy eliminated all gastrointestinal contractile and myoelectric events but antral ECA frequency slowing. Spontaneous occurrences of the myoelectric correlates of vomiting were not different from those activated by apomorphine. These results suggest that ECA disruption may be important for retrograde propagation of the RGC. The function of ECA frequency slowing, however, remains unknown.


Retrieval from episodic memory: neural mechanisms of interference resolution.
Selectively retrieving a target memory among related memories requires some degree of inhibitory control over interfering and competing memories, a process assumed to be supported by inhibitory mechanisms. Evidence from behavioral studies suggests that such inhibitory control can lead to subsequent forgetting of the interfering information, a finding called retrieval-induced forgetting [Anderson, M. C., Bjork, R. A., & Bjork, E. L. Remembering can cause forgetting: Retrieval dynamics in long-term memory. Journal of Experimental Psychology: Learning, Memory & Cognition, 20, 1063-1087, 1994]. In the present functional magnetic resonance imaging study, we investigated the neural processes underlying retrieval-induced forgetting and, in particular, examined the extent to which these processes are retrieval (i.e., selection) specific. Participants actively retrieved a subset of previously studied material (selection condition), or were re-exposed to the same material for relearning (nonselection condition). Replicating prior behavioral work, selective retrieval caused significant forgetting of the nonretrieved items on a delayed recall test, relative to the re-exposure condition. Selective retrieval was associated with increased BOLD responses in the posterior temporal and parietal association cortices, in the bilateral hippocampus, and in the dorsolateral prefrontal cortex. Medial and lateral prefrontal areas showed a strong negative linear relationship between selection-related neural activity and subsequent forgetting of competitors. These findings suggest reduced demands on inhibitory control processes when interference is successfully resolved during early selective retrieval from episodic memory.


Disparities in outcomes after spine surgery: a Michigan Spine Surgery Improvement Collaborative study.
Most studies on racial disparities in spine surgery lack data granularity to control for both comorbidities and self-assessment metrics. Analyses from large, multicenter surgical registries can provide an enhanced platform for understanding different factors that influence outcome. In this study, the authors aimed to determine the effects of race on outcomes after lumbar surgery, using patient-reported outcomes (PROs) in 3 areas: the North American Spine Society patient satisfaction index, the minimal clinically important difference (MCID) on the Oswestry Disability Index (ODI) for low-back pain, and return to work. The Michigan Spine Surgery Improvement Collaborative was queried for all elective lumbar operations. Patient race/ethnicity was categorized as Caucasian, African American, and "other." Measures of association between race and PROs were calculated with generalized estimating equations (GEEs) to report adjusted risk ratios. The African American cohort consisted of a greater proportion of women with the highest comorbidity burden. Among the 7980 and 4222 patients followed up at 1 and 2 years postoperatively, respectively, African American patients experienced the lowest rates of satisfaction, MCID on ODI, and return to work. Following a GEE, African American race decreased the probability of satisfaction at both 1 and 2 years postoperatively. Race did not affect return to work or achieving MCID on the ODI. The variable of greatest association with all 3 PROs at both follow-up times was postoperative depression. While a complex myriad of socioeconomic factors interplay between race and surgical success, the authors identified modifiable risk factors, specifically depression, that may improve PROs among African American patients after elective lumbar spine surgery.


Orientation of the heterodimeric aryl hydrocarbon (dioxin) receptor complex on its asymmetric DNA recognition sequence.
The 2,3,7,8-tetrachlorodibenzo-p-dioxin-transformed aryl hydrocarbon receptor (AHR) complex binds to xenobiotic-responsive element (XRE) sequences in the 5' flanking region of the CYP1A1 gene, resulting in initiation of transcription. Both components of the transformed AHR complex [the ligand-binding AHR monomer and the AHR nuclear translocator (ARNT)] directly contact the XRE. These proteins belong to a novel subclass of basic helix-loop-helix transcription factors. The binding sites of AHR and ARNT on the asymmetric XRE were determined using nuclear extracts of 2,3,7,8-tetrachlorodibenzo-p-dioxin-treated Hepa-1c1c7 cells and a panel of double-stranded oligonucleotides containing XRE1 of the CYP1A1 gene (5'-TTGCGTGAGAA-3'), in which all combinations of three, two, or one of the thymines indicated were substituted by the photoreactive thymine analog 5-bromodeoxyuracil. Covalent cross-linking analysis and immunoprecipitation with antibodies specific for AHR or ARNT demonstrated that ARNT directly contacts the 3'-most thymine position, that AHR directly contacts the second thymine position, and that neither protein contacts the 5'-most thymine position. The thymine position contacted by ARNT lies within a three-nucleotide sequence (5'-GTG-3') identical to a half-site of an E-box element (5'-CACGTG-3') that is recognized by a number of other basic helix-loop-helix transcription factors. AHR binds to a portion of the XRE that does not resemble an E-box. Additional experiments demonstrated that neither protein loops over to contact residues located beyond the other's binding site.


DsbA-L alleviates endoplasmic reticulum stress-induced adiponectin downregulation.
Obesity impairs adiponectin expression, assembly, and secretion, yet the underlying mechanisms remain elusive. The aims of this study were 1) to determine the molecular mechanisms by which obesity impairs adiponectin multimerization and stability, and 2) to determine the potential role of disulfide-bond-A oxidoreductase-like protein (DsbA-L), a recently identified adiponectin interactive protein that promotes adiponectin multimerization and stability in obesity-induced endoplasmic reticulum (ER) stress and adiponectin downregulation. Tauroursodeoxycholic acid (TUDCA), a chemical chaperone that alleviates ER stress, was used to study the mechanism underlying obesity-induced adiponectin downregulation in db/db mice, high-fat diet-induced obese mice, and in ER-stressed 3T3-L1 adipocytes. The cellular levels of DsbA-L were altered by RNAi-mediated suppression or adenovirus-mediated overexpression. The protective role of DsbA-L in obesity- and ER stress-induced adiponectin downregulation was characterized. Treating db/db mice and diet-induced obese mice with TUDCA increased the cellular and serum levels of adiponectin. In addition, inducing ER stress is sufficient to downregulate adiponectin levels in 3T3-L1 adipocytes, which could be protected by treating cells with the autophagy inhibitor 3-methyladenine or by overexpression of DsbA-L. ER stress plays a key role in obesity-induced adiponectin downregulation. In addition, DsbA-L facilitates adiponectin folding and assembly and provides a protective effect against ER stress-mediated adiponectin downregulation in obesity.


Elaborating on systems thinking in health promotion practice.
Health and well-being are the result of a series of complex processes in which an individual interacts with other people and the environment. A systematic approach ensures incorporation of individual, ecological, social and political factors. However, interactions between these factors can be overlooked within a systematical approach. A systemic approach can provide additional information by incorporating interactions and communication. The opportunities of a systems thinking perspective for health promotion were investigated for this paper. Although others have also made attempts to explore systems thinking in the field of health promotion, the implications of systems thinking in practice need attention. Other fields such as agricultural extension studies, organizational studies and development studies provide useful experiences with the use of a systems thinking perspective in practice. Building on experiences from these fields, we give a theoretical background in which processes of social learning and innovation play an important role. From this background, we derive an overview of important concepts for the practical application of a systems thinking perspective. These concepts are the structure of the system, meanings attached to actions, and power relations between actors. To make these concepts more explicit and reduce the theoretical character of systems thinking, we use an illustration to elaborate on these concepts in practice. For this purpose, we describe a health promotion partnership in The Netherlands using the concepts structure, meaning and power relations. We show how a systems perspective increases insight in the functioning of a partnership and how this can facilitate processes of social learning and innovation. This article concludes by identifying future opportunities and challenges in adopting systems thinking for health promotion practice. A systems perspective towards health promotion can help projects reaching a more integral and sustainable approach in which the complex nature of health promotion processes is supported. Practical applications of systems thinking are necessary to adapt this perspective.


Early childhood cognitive development is affected by interactions among illness, diet, enteropathogens and the home environment: findings from the MAL-ED birth cohort study.
Millions of children in low-income and middle-income countries (LMICs) are at risk of not reaching their full cognitive potential. Malnutrition and enteric infections in early life are implicated as risk factors; however, most studies on these risks and their associations with cognitive development have failed to adequately account for confounding factors or the accumulation of putative insults. Here, we examine the interaction between infections and illness on cognitive development in LMIC community settings. As part of the Etiology, Risk Factors, and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development (MAL-ED) longitudinal birth cohort study, children from eight LMICs were followed from birth to 24 months to understand the influence of repeated enteric infections on child growth and development. Here, data from six sites were employed to evaluate associations between infection, illness, the home environment, micronutrient intake and status, maternal reasoning, and cognitive development at 24 months. Higher rates of enteropathogen detection and days with illness were associated with lower haemoglobin concentrations, which in turn were associated with lower cognitive scores at 24 months. Children with lower environmental health/safety scores and lower intakes of vitamin B6 and folate had more enteropathogen detections and illness. Strength of associations varied by weight-for-age in the first 17 days of life; lower weight infants were more susceptible to the negative effects of enteropathogens and illness. Enteropathogens were negatively related to child cognitive development. However, other factors were more strongly associated with child cognition. Targeting of interventions to improve cognitive development should include a focus on reducing frequency of illness, improving the safety and healthfulness of the child's environment, and improving dietary intake.


Electrocatalytic and photocatalytic hydrogen production from acidic and neutral-pH aqueous solutions using iron phosphide nanoparticles.
Nanostructured transition-metal phosphides have recently emerged as Earth-abundant alternatives to platinum for catalyzing the hydrogen-evolution reaction (HER), which is central to several clean energy technologies because it produces molecular hydrogen through the electrochemical reduction of water. Iron-based catalysts are very attractive targets because iron is the most abundant and least expensive transition metal. We report herein that iron phosphide (FeP), synthesized as nanoparticles having a uniform, hollow morphology, exhibits among the highest HER activities reported to date in both acidic and neutral-pH aqueous solutions. As an electrocatalyst operating at a current density of -10 mA cm(-2), FeP nanoparticles deposited at a mass loading of ∼1 mg cm(-2) on Ti substrates exhibited overpotentials of -50 mV in 0.50 M H2SO4 and -102 mV in 1.0 M phosphate buffered saline. The FeP nanoparticles supported sustained hydrogen production with essentially quantitative faradaic yields for extended time periods under galvanostatic control. Under UV illumination in both acidic and neutral-pH solutions, FeP nanoparticles deposited on TiO2 produced H2 at rates and amounts that begin to approach those of Pt/TiO2. FeP therefore is a highly Earth-abundant material for efficiently facilitating the HER both electrocatalytically and photocatalytically.


Dentine matrix protein 1 (DMP-1) is a marker of bone-forming tumours.
Dentin matrix protein 1 (DMP-1) is highly expressed by osteocytes and is a non-collagenous matrix protein found in dentin and bone. In this study, we determined the expression of DMP-1 in mature and immature human bone and examined whether DMP-1 is useful in distinguishing osteoid/bone-forming tumours from other primary and secondary bone tumours. DMP-1 expression was immunohistochemically determined in paraffin sections of a wide range of benign and malignant primary bone tumours and tumour-like lesions (n = 353). DMP-1 mRNA expression was also examined in osteosarcoma and fibrosarcoma cell lines as well as bone tumour specimens (n = 5) using real-time PCR. In lamellar and woven bone, DMP-1 was expressed in the matrix around osteocyte lacunae and canaliculi; osteoblasts and other cell types in the bone were negative. Matrix staining of the osteoid and bone was seen in bone-forming tumours including osteoma, osteoid osteoma, osteoblastoma and osteosarcoma. DMP-1 staining was also seen in fibrous dysplasia, osteofibrous dysplasia and chondroblastoma and in reactive bone in solitary bone cysts and aneurysmal bone cysts. DMP-1 was not expressed in the tumour component of other bone neoplasms including Ewing sarcoma, chondrosarcoma, leiomyosarcoma, fibrosarcoma, giant cell tumour of bone and metastatic carcinoma. DMP-1 mRNA was expressed in osteosarcoma cell lines and tumour samples. DMP-1 is a matrix marker expressed around osteocytes in human woven and lamellar bone and is useful in identifying osteosarcoma and other bone-forming tumours.


Mercy Pregnancy and Emotional Well-being Study (MPEWS): Understanding maternal mental health, fetal programming and child development. Study design and cohort profile.
Maternal mental health represents a significant global health burden. The Mercy Pregnancy and Emotional Well-being Study (MPEWS) was established to provide a comprehensive investigation of early developmental mechanisms and modifiers for maternal, fetal and child emotional well-being. MPEWS is a prospective, longitudinal study from pregnancy to 36 months postpartum that includes diagnostic measures of maternal mental health, observational measures of the mother-infant relationship, measures of child development, and repeat biological sampling. A total of 282 pregnant women were recruited in early pregnancy from the Mercy Hospital for Women in Melbourne, Australia, including 52 women on antidepressant medication, 31 non-medicated women meeting diagnostic criteria for current unipolar depression or dysthymia, and 65 women with a past history of depression. Sample recruitment characteristics included a mean age of 31 years and average gestation of 16 weeks. The MPEWS cohort was comparable to national averages for Australia on key pregnancy and birth variables. Those participants taking antidepressant medication had higher mean Edinburgh Postnatal Depression Scale (EPDS) and State Trait Anxiety Inventory (STAI) scores than the cohort as a whole but were comparable on other key variables. The MPEWS protocol provides a unique opportunity to evaluate the impact of pregnancy mental health on future maternal mental health and child development to aid the development of evidence-based interventions. The study is open for collaborative proposals via approach to the principal investigators.


Serum α-Hydroxybutyrate: A Candidate Marker of Insulin Resistance Is Associated with Deterioration in Anthropometric Measurements in Individuals with Low Diabetes Risk.
α-Hydroxybutyrate (α-HB) is a marker of insulin resistance (IR) and lipid oxidation, both of which precede the development of diabetes and cardiovascular disorders. We aimed to analyze the relation of α-HB levels with anthropometric measurements in individuals without metabolic risk factors. A total of 82 nonobese individuals [body mass index (BMI) <30 kg/m2] without an accompanying chronic disorder were enrolled into the study. The entire cohort of participants underwent physical examination. Biochemical and hormonal parameters were analyzed. The BMI was calculated as weight/height2 (kg/m2). An ELISA method was used to analyze serum α-HB level. The relation of variables was analyzed by correlation analysis. The mean age, BMI, body fat ratio, and waist/hip ratio of participants were 36 (9) years, 24.9 (2.2), 39.2 (3.9), and 0.82 (0.06), respectively. The mean fasting glucose, insulin, homeostasis model assessment of insulin resistance (HOMA-IR) levels, total cholesterol, triglyceride, HDL, and LDL levels were 90.7 (5.1) mg/dL, 9.8 (1.5) IU/mL, 2.2 (0.3), 193.2 (32.6) mg/dL, 119.3 (60.3) mg/dL, 54.6 (12.2) mg/dL, and 114.2 (30.4) mg/dL, respectively. Serum α-HB level was significantly correlated with age, BMI, body fat ratio, waist circumference, waist/hip ratio, fasting glucose, insulin, HOMA-IR, HDL, total cholesterol, and triglyceride. Serum α-HB, a strong marker of insulin resistance, is well correlated with deterioration of anthropometric parameters such as an increase in BMI and body fat distribution in patients with low diabetes risk.


A new designer drug 5F-ADB activates midbrain dopaminergic neurons but not serotonergic neurons.
N-[[1-(5-fluoropentyl)-1H-indazol-3-yl]carbonyl]-3-methyl-D-valine methyl ester (5F-ADB) is one of the most potent synthetic cannabinoids and elicits severe psychotic symptoms in humans, sometimes causing death. To investigate the neuronal mechanisms underlying its toxicity, we examined the effects of 5F-ADB on midbrain dopaminergic and serotonergic systems, which modulate various basic brain functions such as those in reward-related behavior. 5F-ADB-induced changes in spontaneous firing activity of dopaminergic and serotonergic neurons were recorded by ex vivo electrophysiological techniques. In dopaminergic neurons, 5F-ADB (1 μM) significantly increased the spontaneous firing rate, while 5F-ADB failed to activate dopaminergic neurons in the presence of the CB1 antagonist AM251 (1 μM). However, the same concentration of 5F-ADB did not affect serotonergic-neuron activity. These results suggest that 5F-ADB activates local CB1 receptors and potentiates midbrain dopaminergic systems with no direct effects on midbrain serotonergic systems.


Factors affecting deer ked (Lipoptena cervi) prevalence and infestation intensity in moose (Alces alces) in Norway.
The deer ked (Lipoptena cervi), a hematophagous ectoparasite of Cervids, is currently spreading in Scandinavia. In Norway, keds are now invading the south-eastern part of the country and the abundant and widely distributed moose (Alces alces) is the definitive host. However, key factors for ked abundance are poorly elucidated. The objectives of our study were to (i) determine deer ked infestation prevalence and intensity on moose and (ii) evaluate if habitat characteristics and moose population density are determinants of deer ked abundance on moose. In order to identify key factors for deer ked abundance, a total of 350 skin samples from the neck of hunted moose were examined and deer keds counted. Infestation intensity was analyzed in relation to moose age and sex, moose population density and landscape characteristics surrounding the killing site. Deer ked infestation prevalence was 100%, but infestation intensity varied from 0.001 to 1.405 keds/cm². Ked intensity was highest in male yearlings (~1.5 years) and positively associated with longitude and Scots pine (Pinus sylvestris) dominated habitat and negatively associated with bogs and latitude. Moose population density during autumn showed a tendency to be positively associated, while altitude tended to be negatively associated with ked intensity. Deer keds exploit the whole moose population within our study area, but are most prevalent in areas dominated by Scots pine. This is probably a reflection of Scots pine being the preferred winter browse for moose in areas with highest moose densities in winter. Ked intensity decreases towards the northwest and partly with increasing altitude, probably explained by the direction of dispersal and reduced temperature, respectively. Abundant deer ked harm humans and domestic animals. Moose management authorities should therefore be aware of the close relationship between moose, deer ked and habitat, using the knowledge as a management tool for locally regulating the ked burden.


Survey of Neonatal Intensive Care Unit Nurse Attitudes Toward Therapeutic Hypothermia Treatment.
The traumatic experiences of parents of babies treated with therapeutic hypothermia (TH) have been described. No research has assessed neonatal intensive care unit (NICU) nurse experience in providing care to hypothermic babies and emotional support to their parents. To assess NICU nurse attitudes to the provision of TH with respect to perceptions about baby pain/sedation, need for nurse and parent education, decision making about initiation of TH, and barriers to best care. A survey was electronically sent to 219 nurses at 2 affiliated academic level III NICUs: 1 rural and 1 urban location. There were 17 questions where responses were selected from a preset list and 7 opportunities for nurses to provide free text responses. The response rate was 38% (N = 83). Overwhelming similarities between the urban and rural institutions were found with NICU nurses expressing understanding of the indications for initiating TH, agreement that TH improves long-term outcomes and that the benefits of TH outweigh the risks. Nurses at the urban institution more frequently expressed concerns surrounding inadequate treatment of baby pain/sedation, and nurses at both institutions strongly emphasized the need for more nurse and parent education about TH and improved timeliness of decision making for initiation of TH. NICU nurses specifically want to learn more about outcomes of babies after treatment with TH and feel that parents need more education about TH. Research is urgently needed to better understand the implications of TH treatment for parent-baby bonding.


Nitric oxide regulates shear stress-induced early growth response-1. Expression via the extracellular signal-regulated kinase pathway in endothelial cells.
Endothelial cells (ECs) subjected to shear stress constantly release nitric oxide (NO). The effect of NO on shear stress-induced endothelial responses was examined. ECs subjected to shear stress induced a transient and shear force-dependent increase in early growth response-1 (Egr-1) mRNA levels. Treatment of ECs with an NO donor, S-nitroso-N-acetylpenicillamine (SNAP) or 3-morpholinosydnonimine (SIN-1), inhibited this shear stress-induced Egr-1 expression. Conversely, an NO synthase inhibitor to ECs, N(G)-monomethyl-L-arginine, augmented this Egr-1 expression. NO modulation of Egr-1 expression was demonstrated by functional analysis of Egr-1 promoter activity using a chimera containing the Egr-1 promoter region (-698 bp) and reporter gene luciferase. In contrast to the enhanced promoter activity after N(G)-monomethyl-L-arginine treatment, shear stress-induced Egr-1 promoter activity was attenuated after ECs were treated with an NO donor. ECs cotransfected with a dominant negative mutant of Ras (RasN17), Raf-1 (Raf301), or a catalytically inactive mutant of extracellular signal-regulated kinase (ERK)-2 (mERK) inhibited shear stress-induced Egr-1 promoter activity. NO modulation of the signaling pathway was shown by its inhibitory effect on shear stress-induced ERK1/ERK2 phosphorylation and activity. This inhibitory effect was further substantiated by the inhibition of NO on both the shear stress-induced transcriptional activity of Elk-1 (an ERK substrate) and the promoter activity of a reporter construct containing serum response element. NO-treated ECs resulted in a reduction of binding of nuclear proteins to the Egr-1 binding sequences in the platelet-derived growth factor-A promoter region. These results indicate that shear stress-induced Egr-1 expression is modulated by NO via the ERK signaling pathway in ECs. Our findings support the importance of NO as a negative regulator in endothelial responses to hemodynamic forces.


How Attention Changes in Response to Incentives.
Although the performance of simple cognitive tasks can be enhanced if an incentive is provided, the mechanisms enabling such motivational control are not known. This study sought to uncover how mechanisms of attention and readiness are altered by reward-associated incentive stimuli. We measured EEG/ERP activity as human adults viewed a high- or low-incentive cue, experienced a short preparation interval, and then performed a simple visual search task to gain the predicted reward. Search performance was faster with high versus low incentives, and this was accompanied by distinct incentive-related EEG/ERP patterns at each phase of the task (incentive, preparation, and search). First, and most surprisingly, attention to high but not low incentive cues was actively suppressed, as indexed by a PD component in response to the incentive display. During the subsequent preparation interval, neural oscillations in the alpha frequency range were reduced after high-incentive cues, indicating heightened visual readiness. Finally, attentional orienting to the target in the search array was deployed with relatively little effort on high-incentive trials, as indexed by a reduced N2pc component. These results reveal the chain of events by which the brain's executive control mechanisms respond to incentives by altering the operation of multiple processing systems to produce optimal performance.


The Plant Genome Integrative Explorer Resource: PlantGenIE.org.
Accessing and exploring large-scale genomics data sets remains a significant challenge to researchers without specialist bioinformatics training. We present the integrated PlantGenIE.org platform for exploration of Populus, conifer and Arabidopsis genomics data, which includes expression networks and associated visualization tools. Standard features of a model organism database are provided, including genome browsers, gene list annotation, Blast homology searches and gene information pages. Community annotation updating is supported via integration of WebApollo. We have produced an RNA-sequencing (RNA-Seq) expression atlas for Populus tremula and have integrated these data within the expression tools. An updated version of the ComPlEx resource for performing comparative plant expression analyses of gene coexpression network conservation between species has also been integrated. The PlantGenIE.org platform provides intuitive access to large-scale and genome-wide genomics data from model forest tree species, facilitating both community contributions to annotation improvement and tools supporting use of the included data resources to inform biological insight.


[Endocrine profile of patients with post-pill amenorrhea].
"Post-pill amenorrhea" is still a much-debated subject in gynecological endocrinology. The authors therefore studied the reproductive history and endocrinological parameters of 145 patients with the presumptive diagnosis of post-pill amenorrhea, seen in their department during the last two years; 166 cases of secondary amenorrhea served as a control group. The incidence of late menarche (20%), previous oligomenorrhea, and elevated androgen serum levels was high in both groups. There were no significant differences between the two collectives. These findings suggest that although oral contraceptives may have an unfavorable influence on the menstrual cycle, there is no causal relationship between such agents and the development of secondary amenorrhea. It seems that in many cases of presumptive post-pill amenorrhea the common condition of juvenile hyperandrogenemic ovarian insufficiency might be the underlying problem.


Differential Efficacy of Nicotine Replacement Among Overweight and Obese Women Smokers.
Rates of obesity are higher among more dependent smokers and 37%-65% of smokers seeking cessation treatment are overweight or obese. Overweight or obese smokers may possess metabolic and neurobiological features that contribute to difficulty achieving cessation using front-line nicotine replacement products. Attention to factors that facilitate effective cessation treatment in this vulnerable population is needed to significantly reduce mortality risk among overweight and obese smokers. This secondary analysis of 2 large trials of transdermal nicotine replacement in general medical practices evaluated the hypothesis that higher body mass index (BMI) would moderate the efficacy of the nicotine patch. We examined the potential for gender to further moderate the relationship between BMI and treatment efficacy. In the placebo controlled trial (N = 1,621), 21-mg patch was no more effective than placebo for assisting biochemically verified point prevalence abstinence up to 1 year after quitting for women with higher BMI, but appeared to be effective for men at normal or high BMI (gender × BMI beta = -0.22, p = .004). We did not find differential long-term cessation outcomes among male or female smokers in the 15-mg patch trial (n = 705). However, we observed significantly higher rates of early lapse among women with higher BMI treated with nicotine patch across both trials. These results suggest that increased BMI may affect the efficacy of nicotine patch on reducing risk of early lapse in women. Additional research is needed to explore mechanisms of risk for decreased efficacy of this commonly used cessation aid.


Moulds and their secondary metabolites associated with the fermentation and storage of two cocoa bean hybrids in Nigeria.
Fungi and mycotoxin contamination of cocoa beans during fermentation and storage may constitute a hazard in the cocoa value chain and risk to consumers of its products. In this study, fungal profile and secondary metabolite patterns in two cocoa bean hybrids, F and T series, during fermentation and storage were determined. Additionally, secondary metabolite production by the recovered fungi in the beans was examined in culture media. Fungal isolates spanned six genera and eight species: Aspergillus niger, A. tamarii, Paecilomyces variotii, Penicillium citrinum, Pseudopithomyces palmicola, Simplicillium sp., Talaromyces atroroseus and Talaromyces sp.. In both hybrids, Aspergilli (38%) dominated the other fungi while more than one half of all the fungal isolates were from the beans in storage. Among the diverse secondary metabolites produced in media by the isolates were uncommon compounds, e.g. aspulvinone E produced by A. niger, aspterric acid by P. variotii, scalusamid A and sydowinin A by P. citrinum, norlichexanthone and siccanol by Simplicillium, and fallacinol and orsellinic acid by Talaromyces. The strains of P. citrinum produced up to 372 mg/kg citrinin. Forty-four fungal metabolites were quantified in both bean hybrids across the various processing stages, with about 86% occurring in the fermented beans stored for 30 days. The nephrotoxic citrinin, which was not previously reported in cocoa beans worldwide, was the only mycotoxin found in the fermented beans at overall mean concentration of 368 μg/kg. Additionally, its metabolite, dihydrocitrinone, was detected in fermented and stored beans. Consumption of freshly fermented cocoa beans may result in citrinin exposure. Appropriate fungal and mycotoxin control measures are proposed.


Groucho/TLE/R-esp proteins associate with the nuclear matrix and repress RUNX (CBF(alpha)/AML/PEBP2(alpha)) dependent activation of tissue-specific gene transcription.
The Runt related transcription factors RUNX (AML/CBF(alpha)/PEBP2(alpha)) are key regulators of hematopoiesis and osteogenesis. Using co-transfection experiments with four natural promoters, including those of the osteocalcin (OC), multi drug resistance (MDR), Rous Sarcoma Virus long terminal repeat (LTR), and bone sialoprotein (BSP) genes, we show that each of these promoters responds differently to the forced expression of RUNX proteins. However, the three RUNX subtypes (i.e. AML1, AML2, and AML3) regulate each promoter in a similar manner. Although the OC promoter is activated in a C terminus dependent manner, the MDR, LTR and BSP promoters are repressed by three distinct mechanisms, either independent of or involving the AML C terminus, or requiring only the conserved C-terminal pentapeptide VWRPY. Using yeast two hybrid assays we find that the C terminus of AML1 interacts with a Groucho/TLE/R-esp repressor protein. Co-expression assays reveal that TLE proteins repress AML dependent activation of OC gene transcription. Western and northern blot analyses suggest that TLE expression is regulated reciprocally with the levels of OC gene expression during osteoblast differentiation. Digital immunofluorescence microscopy results show that TLE1 and TLE2 are both associated with the nuclear matrix, and that a significant subset of each colocalizes with AML transcription factors. This co-localization of TLE and AML proteins is lost upon removing the C terminus of AML family members. Our findings indicate that suppression of AML-dependent gene activation by TLE proteins involves functional interactions with the C terminus of AML at the nuclear matrix in situ. Our data are consistent with the concept that the C termini of AML proteins support activation or repression of cell-type specific genes depending on the regulatory organization of the target promoter and subnuclear localization.


Arbitrary waveform modulated pulse EPR at 200GHz.
We report here on the implementation of arbitrary waveform generation (AWG) capabilities at ∼200GHz into an Electron Paramagnetic Resonance (EPR) and Dynamic Nuclear Polarization (DNP) instrument platform operating at 7T. This is achieved with the integration of a 1GHz, 2 channel, digital to analog converter (DAC) board that enables the generation of coherent arbitrary waveforms at Ku-band frequencies with 1ns resolution into an existing architecture of a solid state amplifier multiplier chain (AMC). This allows for the generation of arbitrary phase- and amplitude-modulated waveforms at 200GHz with >150mW power. We find that the non-linearity of the AMC poses significant difficulties in generating amplitude-modulated pulses at 200GHz. We demonstrate that in the power-limited regime of ω1<1MHz phase-modulated pulses were sufficient to achieve significant improvements in broadband (>10MHz) spin manipulation in incoherent (inversion), as well as coherent (echo formation) experiments. Highlights include the improvement by one order of magnitude in inversion bandwidth compared to that of conventional rectangular pulses, as well as a factor of two in improvement in the refocused echo intensity at 200GHz.


Insufficient sleep predicts clinical burnout.
The present prospective study aimed to identify risk factors for subsequent clinical burnout. Three hundred eighty-eight working individuals completed a baseline questionnaire regarding work stress, sleep, mood, health, and so forth. During a 2-year period, 15 subjects (7 women and 8 men) of the total sample were identified as "burnout cases," as they were assessed and referred to treatment for clinical burnout. Questionnaire data from the baseline measurement were used as independent variables in a series of logistic regression analyses to predict clinical burnout. The results identified "too little sleep (< 6 h)" as the main risk factor for burnout development, with adjustment for "work demands," "thoughts of work during leisure time," and "sleep quality." The first two factors were significant predictors in earlier steps of the multivariate regression. The results indicate that insufficient sleep, preoccupation with thoughts of work during leisure time, and high work demands are risk factors for subsequent burnout. The results suggest a chain of causation.


Supramolecular photosensitizers rejuvenate photodynamic therapy.
Owing to its spatiotemporal selectivity and noninvasive nature, photodynamic therapy (PDT) has become a clinically promising approach for the treatment of a wide range of cancers and other diseases. However, the full potential of PDT has not been achieved thus far as a consequence of the lack of optimal photosensitizers (PSs) and/or smart transport/activation strategies. These problems, which unfortunately lie at the core of the PDT paradigm, include the oxygen reliance limits, the effect of PDT on hypoxic tumors, limitations of light penetration, and undesired skin photosensitization induced by "always on" PSs. Recently, supramolecular approaches, which rely on the use of non-covalent interactions to construct biomedical active materials, have become suitable methods for developing innovative PSs. Non-covalent interactions enable supramolecular PSs to have sensitive and controllable photoactivities, important elements needed to maximize photodynamic effects and minimize side effects. In addition, versatile supramolecular PS-assemblies can be designed so that PDT occurs synergistically with other therapeutic modalities, e.g., photothermal therapy, leading to a potential improvement of therapeutic effectiveness. In this review, recent progress made in the development of supramolecular PSs for rejuvenating PDT will be presented. Importantly, this discussion also provides a view of future advances that will likely be made in this area and their potential clinical applications.


Evolutionarily conserved interaction between the phosphoproteins and X proteins of bornaviruses from different vertebrate species.
Bornavirus, a non-segmented, negative-strand RNA viruses, is currently classified into several genetically distinct genotypes, such as Borna disease virus (BDV) and avian bornaviruses (ABVs). Recent studies revealed that bornavirus genotypes show unique sequence variability in the putative 5' untranslated region (5' UTR) of X/P mRNA, a bicistronic mRNA for the X protein and phosphoprotein (P). In this study, to understand the evolutionary relationship among the bornavirus genotypes, we investigated the functional interaction between the X and P proteins of four bornavirus genotypes, BDV, ABV genotype 4 and 5 and reptile bornavirus (RBV), the putative 5' UTRs of which exhibit variation in the length. Immunofluorescence and immunoprecipitation analyses using mammalian and avian cell lines revealed that the X proteins of bornaviruses conserve the ability to facilitate the export of P from the nucleus to the cytoplasm via interaction with P. Furthermore, we showed that inter-genotypic interactions may occur between X and P among the genotypes, except for X of RBV. In addition, a BDV minireplicon assay demonstrated that the X and P proteins of ABVs, but not RBV, can affect the polymerase activity of BDV. This study demonstrates that bornaviruses may have conserved the fundamental function of a regulatory protein during their evolution, whereas RBV has evolved distinctly from the other bornavirus genotypes.


A New Illusion at Your Elbow.
On experiencing distal-proximal tactile motion on the volar side of the forearm starting at the wrist, subjects significantly anticipate touch of the elbow crook. This illusion, popular as a children's game, was quantified in ninety participants (forty-seven women) on both arms. As a top-down explanation of the illusion, we discuss a model of Bayesian inferences. As a bottom-up contribution, we consider afterdischarges of cortical neurons, which receive input from skin mechanoreceptors specifically driven by slow-motion tactile stimuli. Like previously described illusions, the elbow crook illusion is larger on the nondominant arm. Women showed a smaller illusion than men, giving testimony to their reportedly superior cutaneous sensitivity.


Ion-selective electrodes based on a pyridyl-containing triazolophane: altering halide selectivity by combining dipole-promoted cooperativity with hydrogen bonding.
Triazolophanes are ionophores, with preorganized cyclic cavities that have tunable selectivities for halides. The interaction with halides is based on hydrogen bonding between the eight CH hydrogen atoms of the cavity and the halide anion. The rigidity of the cavity in tetraphenylene triazolophane along with the hydrogen bonding favors planar 1:1 complexation of "snugly" encapsulated chloride and bromide. Manipulating the triazolophane's structure by introducing two pyridyl moieties into the cavity alters the receptor's binding mode. This change adds a dipole-promoted driving force that combines with hydrogen bonding to favor the formation of 2:1 sandwich complexes around halides. The potentiometric response of electrodes based on this new ionophore was evaluated for optimal halide selectivity. The new triazolophane-based electrode showed an anti-Hofmeister selectivity toward iodide with a submicromolar detection limit. The stoichiometry of complexation and the stability constants with different halides were evaluated using a segmented sandwich membranes method. The pyridyl-triazolophane demonstrated a response consistent with a 2:1 sandwich-type complex with iodide, in polyvinyl chloride (PVC) membranes.


Growing Bayesian network models of gene networks from seed genes.
For the last few years, Bayesian networks (BNs) have received increasing attention from the computational biology community as models of gene networks, though learning them from gene-expression data is problematic. Most gene-expression databases contain measurements for thousands of genes, but the existing algorithms for learning BNs from data do not scale to such high-dimensional databases. This means that the user has to decide in advance which genes are included in the learning process, typically no more than a few hundreds, and which genes are excluded from it. This is not a trivial decision. We propose an alternative approach to overcome this problem. We propose a new algorithm for learning BN models of gene networks from gene-expression data. Our algorithm receives a seed gene S and a positive integer R from the user, and returns a BN for the genes that depend on S such that less than R other genes mediate the dependency. Our algorithm grows the BN, which initially only contains S, by repeating the following step R + 1 times and, then, pruning some genes; find the parents and children of all the genes in the BN and add them to it. Intuitively, our algorithm provides the user with a window of radius R around S to look at the BN model of a gene network without having to exclude any gene in advance. We prove that our algorithm is correct under the faithfulness assumption. We evaluate our algorithm on simulated and biological data (Rosetta compendium) with satisfactory results.