Thermal soaring flight of birds and unmanned aerial vehicles.
Thermal soaring saves much energy, but flying large distances in this form represents a great challenge for birds, people and unmanned aerial vehicles (UAVs). The solution is to make use of the so-called thermals, which are localized, warmer regions in the atmosphere moving upward with a speed exceeding the descent rate of birds and planes. Saving energy by exploiting the environment more efficiently is an important possibility for autonomous UAVs as well. Successful control strategies have been developed recently for UAVs in simulations and in real applications. This paper first presents an overview of our knowledge of the soaring flight and strategy of birds, followed by a discussion of control strategies that have been developed for soaring UAVs both in simulations and applications on real platforms. To improve the accuracy of the simulation of thermal exploitation strategies we propose a method to take into account the effect of turbulence. Finally, we propose a new GPS-independent control strategy for exploiting thermal updrafts.


ULTRASOUND SPECTROSCOPY.
We introduce the concept of "Ultrasound Spectroscopy". The premise of ultrasound spectroscopy is that by acquiring ultrasound RF data at multiple power and frequency settings, a rich set of features can be extracted from that RF data and used to characterize the underlying tissues. This is beneficial for a variety of problems, such as accurate tissue classification, application-specific image generation, and numerous other quantitative tasks. These capabilities are particularly relevant to point-of-care ultrasound (POCUS) applications, where operator experience with ultrasound may be limited. Instead of displaying B-mode images, a POCUS application using ultrasound spectroscopy can, for example, automatically detect internal abdominal bleeding. In this paper, we present ex vivo tissue phantom studies to demonstrate the accuracy of ultrasound spectroscopy over previous approaches. Our studies suggest that ultrasound spectroscopy provides exceptional accuracy and informative features for classifying blood versus other tissues across image locations and body habitus.


The palliative care knowledge of nursing home staff: The EU FP7 PACE cross-sectional survey in 322 nursing homes in six European countries.
The provision of high-quality palliative care in nursing homes (NHs) is a major challenge and places demands on the knowledge and skills of the staff. This study assesses the palliative care knowledge of staff in NHs in Europe. Cross-sectional study using structured survey Setting/participants: Nurses and care assistants working in 322 representative samples of NHs in Belgium, the Netherlands, England, Finland, Poland and Italy. Palliative care knowledge is measured with the Palliative Care Survey. Scores on the scales range between 0 and 1; higher scores indicate more knowledge. A total of 3392 NH-staff were given a questionnaire, and 2275 responded (67%). Knowledge of basic palliative care issues ranged between 0.20 in Poland (95% confidence interval (CI) 0.19; 0.24) and 0.61 in Belgium (95% CI 0.59; 0.63), knowledge of physical aspects that can contribute to pain ranged between 0.81 in Poland (95% CI 0.79; 0.84) and 0.91 in the Netherlands (95% CI 0.89; 0.93), and knowledge of psychological reasons that can contribute to pain ranged between 0.56 in England (95% CI 0.50; 0.62) and 0.87 in Finland (95% CI 0.83; 0.90). Factors associated with knowledge were country, professional role and having undertaken formal training in palliative care. Knowledge of nurses and care assistants concerning basic palliative care issues appears to be suboptimal in all participating countries, although there is substantial heterogeneity. Education of nursing staff needs to be improved across, but each country may require its own strategy to address the unique and specific knowledge gaps.


Characterization of novel non-nucleoside reverse transcriptase (RT) inhibitor resistance mutations at residues 132 and 135 in the 51 kDa subunit of HIV-1 RT.
Several rare and novel NNRTI [non-nucleoside reverse transcriptase (RT) inhibitor] resistance mutations were recently detected at codons 132 and 135 in RTs from clinical isolates using the yeast-based chimaeric TyHRT (Ty1/HIV-1 RT) phenotypic assay. Ile132 and Ile135 form part of the beta7-beta8 loop of HIV-1 RT (residues 132-140). To elucidate the contribution of these residues in RT structure-function and drug resistance, we constructed twelve recombinant enzymes harbouring mutations at codons 132 and 135-140. Several of the mutant enzymes exhibited reduced DNA polymerase activities. Using the yeast two-hybrid assay for HIV-1 RT dimerization we show that in some instances this decrease in enzyme activity could be attributed to the mutations, in the context of the 51 kDa subunit of HIV-1 RT, disrupting the subunit-subunit interactions of the enzyme. Drug resistance analyses using purified RT, the TyHRT assay and antiviral assays demonstrated that the I132M mutation conferred high-level resistance (>10-fold) to nevirapine and delavirdine and low-level resistance (approximately 2-3-fold) to efavirenz. The I135A and I135M mutations also conferred low level NNRTI resistance (approximately 2-fold). Subunit selective mutagenesis studies again demonstrated that resistance was conferred via the p51 subunit of HIV-1 RT. Taken together, our results highlight a specific role of residues 132 and 135 in NNRTI resistance and a general role for residues in the beta7-beta8 loop in the stability of HIV-1 RT.


The Influence of Age and Sex on Genetic Associations with Adult Body Size and Shape: A Large-Scale Genome-Wide Interaction Study.
Genome-wide association studies (GWAS) have identified more than 100 genetic variants contributing to BMI, a measure of body size, or waist-to-hip ratio (adjusted for BMI, WHRadjBMI), a measure of body shape. Body size and shape change as people grow older and these changes differ substantially between men and women. To systematically screen for age- and/or sex-specific effects of genetic variants on BMI and WHRadjBMI, we performed meta-analyses of 114 studies (up to 320,485 individuals of European descent) with genome-wide chip and/or Metabochip data by the Genetic Investigation of Anthropometric Traits (GIANT) Consortium. Each study tested the association of up to ~2.8M SNPs with BMI and WHRadjBMI in four strata (men ≤50y, men >50y, women ≤50y, women >50y) and summary statistics were combined in stratum-specific meta-analyses. We then screened for variants that showed age-specific effects (G x AGE), sex-specific effects (G x SEX) or age-specific effects that differed between men and women (G x AGE x SEX). For BMI, we identified 15 loci (11 previously established for main effects, four novel) that showed significant (FDR<5%) age-specific effects, of which 11 had larger effects in younger (<50y) than in older adults (≥50y). No sex-dependent effects were identified for BMI. For WHRadjBMI, we identified 44 loci (27 previously established for main effects, 17 novel) with sex-specific effects, of which 28 showed larger effects in women than in men, five showed larger effects in men than in women, and 11 showed opposite effects between sexes. No age-dependent effects were identified for WHRadjBMI. This is the first genome-wide interaction meta-analysis to report convincing evidence of age-dependent genetic effects on BMI. In addition, we confirm the sex-specificity of genetic effects on WHRadjBMI. These results may provide further insights into the biology that underlies weight change with age or the sexually dimorphism of body shape.


The Association of Serum Testosterone Levels With Recurrence and Mortality After Acute Ischemic Stroke in Males.
The current study aimed to investigate whether low testosterone predicted the recurrence and clinical outcomes after acute ischemic stroke (AIS) in males. From June 2015 through August 2017, the study prospectively enrolled 110 male AIS patients. All received detailed evaluations at admission and were followed for at least 1 year. The cumulative incidence, overall survival, length of hospital stay, and the percentage of previous stroke were compared between subjects with testosterone <440 ng/dl and >440 ng/dl. The median age was 62 years (range, 35-93 years). The median serum testosterone at admission was 438 [203] ng/dl (range, 44-816 ng/dl); 55 patients (50%) had testosterone <440 ng/dl and were considered as low testosterone. The median follow-up was 23 months. During the period, 12 recurrences and 10 deaths occurred. The 1-year and 3-year cumulative recurrence rate were 8.3% and 11.9%, respectively; the 1-year and 3-year overall survival were 96.3% and 84.6%, respectively. The cumulative recurrence rates were similar between the two testosterone groups (log-rank test, p = .88). Low testosterone was associated with poor survival with marginal significance (log-rank test, p = .079). Men with low testosterone had a higher percentage of previous stroke (29.1% versus 12.7%, p = .035). The mean lengths of hospital stay were similar between the two testosterone groups (16.6 ± 15.8 days versus 14.0 ± 10.6, p = .31). Total testosterone at admission fails to predict stroke recurrence. However, men with low testosterone at admission are more likely to have previous stroke and may have a higher all-cause mortality rate after AIS.


Matching behavior and the representation of value in the parietal cortex.
Psychologists and economists have long appreciated the contribution of reward history and expectation to decision-making. Yet we know little about how specific histories of choice and reward lead to an internal representation of the "value" of possible actions. We approached this problem through an integrated application of behavioral, computational, and physiological techniques. Monkeys were placed in a dynamic foraging environment in which they had to track the changing values of alternative choices through time. In this context, the monkeys' foraging behavior provided a window into their subjective valuation. We found that a simple model based on reward history can duplicate this behavior and that neurons in the parietal cortex represent the relative value of competing actions predicted by this model.


Amyloid deposition as a cause of atrial remodelling in persistent valvular atrial fibrillation.
The spectrum of histological alterations, namely atrial amyloidosis, in the right and left atria of patients with chronic persistent atrial fibrillation (AF) and rheumatic heart disease is not completely known. One hundred and twenty-eight atrial appendages (66 left and 62 right), obtained from 72 patients with rheumatic valve disease and chronic AF undergoing cardiac surgery for valve replacement or repair and AF treatment were histologically evaluated for the presence of amyloid deposits. One hundred and four specimens of left and right auricles from 52 patients in sinus rhythm with severe chronic heart failure undergoing heart transplant were also analyzed (controls). Amyloid was found in 33 (46%) valvular patients with chronic persistent AF and in 6 (12%) controls. Amyloid was related to the presence and duration of AF, was more frequently found in left atrial samples and was independent of age. On stepwise logistic regression analysis, AF duration and female gender were independently related to amyloid deposition. Patients with long-standing AF and rheumatic heart disease have a very high prevalence of atrial amyloidosis. Amyloid deposition is more frequent in left than in right atrial appendage and correlates with AF duration and female gender. Amyloid deposition could constitute an additional histological feature in the structural remodeling of atria during long-standing AF, at least in rheumatic valve disease. Persistence of AF might play a pivotal role in promoting amyloid deposition.


Colorimetric Humidity and Solvent Recognition Based on a Cation-Exchange Clay Mineral Incorporating Nickel(II)-Chelate Complexes.
Solvatochromic nickel(II) complexes with diketonato and diamine ligands were incorporated into a saponite clay by ion exchange, and their colorimetric humidity- and solvent-recognition properties were investigated. These powders exhibit color change from red to blue-green depending on humidity, and the detection range can be controlled by modifying the metal complex. The humidity response takes advantage of the humidity-dependent water content in clay and the coordination of water molecules to the metal complex in equilibrium. The addition of organic solvents to the powders causes a color change to occur, varying from red to blue-green depending on the donor number of the solvent, thereby enabling solvent recognition. In the clay, the affinity of less sterically hindered complexes to water or solvent molecules is decreased compared with that in solution because the cationic complexes interact with the anionic layers in the clay. Incorporating diethylene glycol into the materials produced thermochromic powders.


A randomized trial examining the effectiveness of switching from olanzapine, quetiapine, or risperidone to aripiprazole to reduce metabolic risk: comparison of antipsychotics for metabolic problems (CAMP).
The authors conducted a multisite randomized controlled trial examining the strategy of switching from olanzapine, quetiapine, or risperidone to aripiprazole to ameliorate metabolic risk factors for cardiovascular disease. Patients with schizophrenia or schizoaffective disorder with a body mass index ≥ 27 and non-high-density lipoprotein (non-HDL) cholesterol ≥ 130 mg/dl who were on a stable treatment dosage of olanzapine, quetiapine, or risperidone were randomly assigned to switch to ari-piprazole (N=109) for 24 weeks or stay on their current medication (N=106). All participants were enrolled in a behaviorally oriented diet and exercise program. Clinical raters were blinded to treatment assignment. The primary and key secondary outcomes were change in non-HDL cholesterol and efficacy failure, respectively. The prespecified primary analysis included 89 switchers and 98 stayers who had at least one postbaseline non-HDL cholesterol measurement. The least squares mean estimates of non-HDL cholesterol decreased more for the switch group than for the stay group (-20.2 mg/dl and -10.8 mg/dl, respectively). Switching was associated with larger weight reductions (least squares mean=2.9 kg) and a net reduction of serum triglycerides of 32.7 mg/dl. Twenty-two switchers (20.6%) and 18 stayers (17.0%) experienced protocol-defined efficacy failure. Forty-seven switchers (43.9%) and 26 stayers (24.5%) discontinued the assigned antipsychotic medication before 24 weeks. Switching to aripiprazole led to improvement of non-HDL cholesterol levels and other metabolic parameters. Rates of efficacy failure were similar between groups, but switching to aripiprazole was associated with a higher rate of treatment discontinuation. In the context of close clinical monitoring, switching from an antipsychotic with high metabolic risk to one with lower risk to improve metabolic parameters is an effective strategy.


IL (Interleukin)-10-STAT3-Galectin-3 Axis Is Essential for Osteopontin-Producing Reparative Macrophage Polarization After Myocardial Infarction.
Both osteopontin (OPN) and galectin-3 have been implicated in phagocytic clearance of dead cells and reparative fibrosis during wound healing. CD206+ macrophages are involved in tissue repair through phagocytosis and fibrosis after myocardial infarction (MI). However, the relationship among OPN, galectin-3, and macrophage polarization in the context of MI remains unclear. The time course of Spp1 (encoding OPN) expression in the heart after MI showed a strong activation of Spp1 on day 3 after MI. To identify where in the body and in which cells the transcriptional activity of Spp1 increased after MI, we analyzed EGFP (enhanced green fluorescent protein)- Spp1 knockin reporter mice on day 3 after MI. The transcriptional activity of Spp1 increased only in CD206+ macrophages in the infarct myocardium, and most of CD206+ macrophages have strong transcriptional activation of Spp1 after MI. The temporal expression pattern of Lgal3 (encoding galectin-3) in cardiac macrophages after MI was similar to that of Spp1, and OPN is almost exclusively produced by galectin-3hiCD206+ macrophages. Although both interleukin (IL)-4 and IL-10 were reported to promote CD206+ macrophage-mediated cardiac repair after MI, IL-10- but not IL-4-stimulated CD11b+Ly6G- cells could differentiate into OPN-producing galectin-3hiCD206+ macrophages and showed enhanced phagocytic ability. Inhibition of STAT3 tyrosine phosphorylation suppressed IL-10-induced expression of intracellular galectin-3 and transcriptional activation of Spp1. Knockdown of galectin-3 suppressed their ability to differentiate into OPN-producing cells, but not STAT3 activation. The tyrosine phosphorylation of STAT3 and the appearance rate of galectin-3hiCD206+ cells on cardiac CD11b+Ly6G- cells in Spp1 knockout mice were the same as those in wild-type mice. Spp1 knockout mice showed vulnerability to developing post-MI left ventricular chamber dilatation and the terminal deoxynucleo-tidyltransferase 2'-Deoxyuridine-5'-triphosphate nick-end labeling (TUNEL)-positive cells in the infarcted myocardium after MI remained higher in number in Spp1 knockout mice than in wild-type mice. OPN is almost exclusively produced by galectin-3hiCD206+ macrophages, which specifically appear in the infarct myocardium after MI. The IL-10-STAT3-galectin-3 axis is essential for OPN-producing reparative macrophage polarization after myocardial infarction, and these macrophages contribute to tissue repair by promoting fibrosis and clearance of apoptotic cells. These results suggest that galectin-3 may contribute to reparative fibrosis in the infarct myocardium by controlling OPN levels.


High molecular weight kininogen regulates prekallikrein assembly and activation on endothelial cells: a novel mechanism for contact activation.
The consequences of assembling the contact system of proteins on the surface of vascular cells has received little study. We asked whether assembly of these proteins on the surface of cultured human endothelial cells (HUVECs) results in the activation of prekallikrein (PK) and its dependent pathways. Biotinylated PK binds specifically and reversibly to HUVECs in the presence of high molecular weight kininogen (HK) (apparent Kd of 23 +/- 11 nmol/L, Bmax of 1.7 +/- 0.5 x 10(7) sites per cell [mean +/- SD, n = 5 experiments]). Cell-associated PK is rapidly converted to kallikrein. Surprisingly, the activation of cell-associated HK.PK complexes is entirely independent of exogenous factor XII (Km = 30 nmol/L, Vmax = 12 +/- 3 pmol/L/min in the absence v Km = 20 nmol/L, Vmax = 9.2 +/- 2.1 pmol/L/min in the presence of factor XII). Rather, kallikrein formation is mediated by an endothelial cell-associated, thiol protease. Cell-associated HK is proteolyzed during the course of prekallikrein activation, releasing kallikrein from the surface. Furthermore, activation of PK bound to HK on HUVECs promotes kallikrein-dependent activation of pro-urokinase, resulting in the formation of plasmin. These results indicate the existence of a previously undescribed, factor XII-independent pathway for contact factor activation on HUVECs that regulates the production of bradykinin and may contribute to cell-associated plasminogen activation in vivo.


The reassigned pseudo Wigner-Ville transform in electrochemical noise analysis.
Several different time-frequency transforms from signal processing were used to analyze electrochemical noise data to determine frequency components contained within the noise record and their time evolution. Bilinear time-frequency representations (TFR) based on the Wigner-Ville distribution (WVD) were compared with a special focus on the reassigned smoothed pseudo WVD (RSPWVD). Spectra obtained with WVD were compared with traditional linear time-frequency representations, such as short time Fourier transform and wavelet transform. Comparison to other TFRs showed that the RSPWVD suppressed artifacts, provided better resolution of the time-frequency analysis in both time and frequency domains, and improved the overall readability of a representation. The obtained spectra from RSPWVD were consistent with the results from DWT, but permitted a more comprehensive analysis of transients. Consequently, RSPWVD is suitable for electrochemical noise analysis. In the presence of Cl-, RSPWVD showed that the passivity of Al was compromised, as evidenced by the presence of various current transients in the frequency range from 10-2 to 1 Hz.


The cryo-EM structure of the SNX-BAR Mvp1 tetramer.
Sorting nexins (SNX) are a family of PX domain-containing proteins with pivotal roles in trafficking and signaling. SNX-BARs, which also have a curvature-generating Bin/Amphiphysin/Rvs (BAR) domain, have membrane-remodeling functions, particularly at the endosome. The minimal PX-BAR module is a dimer mediated by BAR-BAR interactions. Many SNX-BAR proteins, however, additionally have low-complexity N-terminal regions of unknown function. Here, we present the cryo-EM structure of the full-length SNX-BAR Mvp1, which is an autoinhibited tetramer. The tetramer is a dimer of dimers, wherein the membrane-interacting BAR surfaces are sequestered and the PX lipid-binding sites are occluded. The N-terminal low-complexity region of Mvp1 is essential for tetramerization. Mvp1 lacking its N-terminus is dimeric and exhibits enhanced membrane association. Membrane binding and remodeling by Mvp1 therefore requires unmasking of the PX and BAR domain lipid-interacting surfaces. This work reveals a tetrameric configuration of a SNX-BAR protein that provides critical insight into SNX-BAR function and regulation.


Nef protein of HIV-1 is a transcriptional repressor of HIV-1 LTR.
In studies of the genetics of human immunodeficiency virus type 1 (HIV-1), the product of the nef gene, formerly known as F, 3'-orf, or B-ORF, was a negative regulator of HIV-1 replication. Proviruses with mutations in the nef gene replicated better than their standard counterparts during transient expression, and the mutant virus maintained its enhanced replication even after serial passages in T lymphocytes. The nef protein trans-suppressed, in a dose-dependent manner, the replication of wild-type and nef mutant proviruses and the expression of reporter genes linked to the HIV-1 long terminal repeat (LTR). The repression induced by the nef protein was mediated by inhibition of transcription from the HIV-1 LTR, which contains a far upstream cis element (previously recognized to be a negative regulatory element) between 340 and 156 nucleotides upstream of the RNA initiation site.


[Modeling familial Alzheimer's disease with induced pluripotent stem cells].
Alzheimer's disease (AD) is the most common form of age-related dementia, characterized by progressive memory loss and cognitive disturbance. According to the amyloid cascade hypothesis, a prevailing theory of AD pathology, accumulation of toxic Aβ42, in the brain is the initiator of AD pathogenesis, subsequently leading to the formation of neurofibrillary tangles, and consequently neuronal loss. Mutations of presenilin 1 (PS1) and presenilin 2 (PS2), which are catalytic components of γ-secretase, are causative factors for autosomal dominant early-onset familial AD (FAD). Induced pluripotent stem cell (iPSC) technology provides a new method for elucidating the molecular basis of human diseases, including neurodegenerative diseases. Here we generate iPSCs from fibroblasts of FAD patients with mutations in PS1 (A246E) and PS2 (N141I), and characterize the differentiation of these cells into neurons. We find that FAD-iPSC-derived differentiated neurons have increased toxic Aβ42 secretion, recapitulating the molecular pathogenesis of mutant presenilins. Furthermore, secretion of Aβ42 from these neurons sharply responds to γ secretase inhibitors and modulators, indicating the potential for identification and validation of candidate drugs. Our findings demonstrate that the FAD-iPSC-derived neuron is a valid model of AD and provides an innovative strategy for the study of late-onset neurodegenerative diseases.


[Respiratory virosis and invasive bacterial superinfections. The case for influenza and meningococcal diseases].
The pathophysiology of bacterial superinfections of influenza, including meningococcal diseases, remains obscure. Mice, normally resistant to the meningococcus, become susceptible after previous influenza A virus infection. This immunosuppressive effect is transitory and is associated with the peak of the inflammatory anti-virus reaction. These results underline the importance of preventing bacterial superinfections of influenza by the surveillance of any relapse of fever after improvement of the influenza syndrome. At the community level, influenza vaccine, beside its specific effects, might also prevent many cases of invasive superinfections, including meningococcal diseases.


Global minimally invasive pyeloplasty study in children: Results from the Pediatric Urology Expert Group of the European Association of Urology Young Academic Urologists working party.
Minimally invasive pyeloplasty (MIP) for ureteropelvic junction (UPJ) obstruction in children has gained popularity over the past decade as an alternative to open surgery. The present study aimed to identify the factors affecting complication rates of MIP in children, and to compare the outcomes of laparoscopic (LP) and robotic-assisted laparoscopic pyeloplasty (RALP). The perioperative data of 783 pediatric patients (<18 years old) from 15 academic centers who underwent either LP or RALP with an Anderson Hynes dismembered pyeloplasty technique were retrospectively evaluated. Redo cases and patients with anatomic renal abnormalities were excluded. Demographics and operative data, including procedural factors, were collected. Complications were classified according to the Satava and modified Clavien systems. Failure was defined as any of the following: obstructive parameters on diuretic renal scintigraphy, decline in renal function, progressive hydronephrosis, or symptom relapse. Univariate and multivariate analysis were applied to identify factors affecting the complication rates. All parameters were compared between LP and RALP. A total of 575 children met the inclusion criteria. Laparoscopy, increased operative time, prolonged hospital stay, ureteral stenting technique, and time required for stenting were factors influencing complication rates on univariate analysis. None of those factors remained significant on multivariate analysis. Mean follow-up was 12.8 ± 9.8 months for RALP and 45.2 ± 33.8 months for LP (P = 0.001). Hospital stay and time for stenting were shorter for robotic pyeloplasty (P < 0.05 for both). Success rates were similar between RALP and LP (99.5% vs 97.3%, P = 0.11). The intraoperative complication rate was comparable between RALP and LP (3.8% vs 7.4%, P = 0.06). However, the postoperative complication rate was significantly higher in the LP group (3.2% for RALP and 7.7% for LP, P = 0.02). All complications were of no greater severity than Satava Grade IIa and Clavien Grade IIIb. This was the largest multicenter series of LP and RALP in the pediatric population. Limitations of the study included the retrospective design and lack of surgical experience as a confounder. Both minimally invasive approaches that were studied were safe and highly effective in treating UPJ obstruction in children in many centers globally. However, shorter hospitalization time and lower postoperative complication rates with RALP were noted. The aims of the study were met.


Motivational reserve: lifetime motivational abilities contribute to cognitive and emotional health in old age.
The authors recently developed the concept of motivational reserve, which implies a set of motivational abilities that provide individuals with resilience to neuropathological damage. This study investigated how lifetime motivational abilities are associated with current cognitive status, mild cognitive impairment, and psychological well-being in old age. A community sample of 147 participants without dementia between 60 and 94 years of age, stratified for age group, sex, and education, completed motivation and well-being questionnaires and cognitive tests. A new procedure was used to estimate their midlife motivational and cognitive abilities on the basis of their main occupation using the Occupational Information Network (O*NET) system. O*NET-estimated motivational abilities predicted cognitive status, psychological well-being, and odds of mild cognitive impairment, even when age, sex, education, and cognitive ability were controlled. Although O*NET-estimated cognitive abilities were not significant predictors, scores on a measure of crystallized intelligence were associated with current cognitive status and odds of mild cognitive impairment. Findings suggest that motivational reserve acts as a protective factor against the manifestation of cognitive impairment and emotional problems in later life.


Activation of the esophagin promoter during esophageal epithelial cell differentiation.
Esophagin is a member of the small proline-rich protein family of cell envelope precursor proteins, which are expressed during squamous cell differentiation. Esophagin is expressed at high levels in normal esophageal epithelium, but its expression is absent from esophageal squamous cell carcinomas and adenocarcinomas. Moreover, loss of esophagin expression is present in areas of dysplasia or normal mucosa adjacent to carcinomas, suggesting that absence of esophagin may constitute a harbinger of early esophageal malignant transformation. A greater understanding of transcriptional control of esophagin may provide valuable insights into esophageal malignancy. Therefore, this study was undertaken in order to isolate and carry out initial characterization of a functional promoter for esophagin. A genomic clone containing esophagin was isolated and sequenced, including 2.7 kb of the esophagin promoter region. Esophagin expression was studied in response to various treatments of primary cultured human esophageal epithelial cells and squamous cell carcinoma cell lines. Calcium was the strongest inducer of the endogenous esophagin promoter, with induction occurring at 12-72 hours. In primary cultured esophageal epithelial cells, a region spanning 116 bp upstream of the transcriptional start site to 45 bp downstream was sufficient to direct low, basal, in vitro esophagin expression. However, responsiveness of primary esophageal cells to calcium required inclusion of promoter elements 1688 bp upstream of the transcriptional start site. Site-directed mutagenesis studies suggested a putative role for C/EBP-beta, OCT-1, and OCT-3 transcription factor binding sites in the minimal promoter region. In conjunction with published human in vivo studies, these data support the hypothesis that esophagin is a biomarker of esophageal squamous cell differentiation and provide an in vitro model to evaluate regulatory factors involved in this differentiation process.


Congenital heart defects and left ventricular non-compaction in males with loss-of-function variants in NONO.
The non-POU domain containing octamer-binding gene (NONO) is located on chromosome Xq13.1 and encodes a member of a small family of RNA-binding and DNA-binding proteins that perform a variety of tasks involved in RNA synthesis, transcriptional regulation and DNA repair. Loss-of-function variants in NONO have been described as a cause of intellectual disability in males but have not been described in association with congenital heart defects or cardiomyopathy. In this article, we seek to further define the phenotypic consequences of NONO depletion in human subjects. We searched a clinical database of over 6000 individuals referred for exome sequencing and over 60 000 individuals referred for CNV analysis. We identified two males with atrial and ventricular septal defects, left ventricular non-compaction (LVNC), developmental delay and intellectual disability, who harboured de novo, loss-of-function variants in NONO. We also identified a male infant with developmental delay, congenital brain anomalies and severe LVNC requiring cardiac transplantation, who inherited a single-gene deletion of NONO from his asymptomatic mother. We conclude that in addition to global developmental delay and intellectual disability, males with loss-of-function variants in NONO may also be predisposed to developing congenital heart defects and LVNC with the penetrance of these cardiac-related problems being influenced by genetic, epigenetic, environmental or stochastic factors. Brain imaging of males with NONO deficiency may reveal structural defects with abnormalities of the corpus callosum being the most common. Although dysmorphic features vary between affected individuals, relative macrocephaly is a common feature.


Continuous positive airway pressure for bronchiolitis in a general paediatric ward; a feasibility study.
Continuous positive airway pressure (CPAP) is commonly used to relieve respiratory distress in infants with bronchiolitis, but has mostly been studied in an intensive care setting. Our prime aim was to evaluate the feasibility of CPAP for infants with bronchiolitis in a general paediatric ward, and secondary to assess capillary PCO2 (cPCO2) levels before and during treatment. From May 1(st) 2008 to April 30(th) 2012, infants with bronchiolitis at Stavanger University Hospital were treated with CPAP in a general paediatric ward, but could be referred to an intensive care unit (ICU) when needed, according to in-house guidelines. Levels of cPCO2 were prospectively registered before the start of CPAP and at approximately 4, 12, 24 and 48 hours of treatment as long as CPAP was given. We had a continuous updating program for the nurses and physicians caring for the infants with CPAP. The study was population based. 672 infants (3.4%) were hospitalized with bronchiolitis. CPAP was initiated in 53 infants (0.3%; 7.9% of infants with bronchiolitis), and was well tolerated in all but three infants. 46 infants were included in the study, the majority of these (n = 33) were treated in the general ward only. These infants had lower cPCO2 before treatment (8.0; 7.7, 8.6)(median; quartiles) than those treated at the ICU (n = 13) (9.3;8.5, 9.9) (p < 0.001). The level of cPCO2 was significantly reduced after 4 h in both groups; 1.1 kPa (paediatric ward) (p < 0.001) and 1.3 kPa (ICU) (p = 0.002). Two infants on the ICU did not respond to CPAP (increasing cPCO2 and severe apnoe) and were given mechanical ventilation, otherwise no side effects were observed in either group treated with CPAP. Treatment with CPAP for infants with bronchiolitis may be feasible in a general paediatric ward, providing sufficient staffing and training, and the possibility of referral to an ICU when needed.


Global adiposity rather than abnormal regional fat distribution characterizes women with polycystic ovary syndrome.
Obesity-related predisposition to polycystic ovary syndrome (PCOS) could reflect overall adiposity and/or regional accumulation of abdominal visceral fat. The objective of the study was to compare distributions of visceral, abdominal sc, and gluteofemoral sc adipose tissue in PCOS cases vs. control women. This was a cross-sectional study. Fat depot measurements from axial magnetic resonance imaging scans taken at anatomically predefined sites were compared between 22 body mass index (BMI)/fat mass-matched pairs of PCOS cases and controls; whole-group comparisons included 50 PCOS cases vs. 28 female controls. All subjects were of UK British/Irish origin. We measured cross-sectional areas of adipose tissue within visceral (mid-L4), abdominal (mid-L4) sc, and gluteofemoral (greater trochanteric and midfemoral) sc fat depots. Other measurements included fat mass, BMI, testosterone, SHBG, and homeostasis model assessment of insulin resistance (a measure of insulin sensitivity). Whole-group analyses were adjusted for fat mass and age. There were no significant differences in fat-depot measurements between BMI/fat mass-matched pairs of PCOS cases and controls: mid-L4 visceral (P=0.40), abdominal sc (P=0.22), gluteal sc (P=0.67), and midfemoral sc (P=0.37) depots. Whole-group comparisons gave similar results after adjustments for fat mass and age. Fasting serum insulin concentrations (P=0.03) and homeostasis model assessment of insulin resistance (P=0.03) were significantly higher in the PCOS group than BMI/fat mass-matched controls. PCOS cases and BMI/fat mass-matched control women are indistinguishable with respect to distribution of fat within visceral, abdominal sc, and gluteofemoral sc depots, despite significant differences in insulin resistance between these two groups.


Nutrition transition in India.
The primary objective of this review is to examine the demographic and nutrition transition in India in relation to its contribution to the emerging epidemic of chronic non-communicable diseases in this country. India, the country as a whole and its different states with a population exceeding 1 billion in 2001. The review examines demographic changes in the population with consequent effects on the population pyramid, the rapidity and rates of urbanisation with striking variations in chronic disease patterns and the trends in obesity between rural and urban communities, attempting to relate their prevalence with the diet and lifestyle changes accompanying them. The review is based largely on representative large-scale surveys in the country and other reliable documented data on population characteristics. It also includes a review of the published literature. The results indicate that the demographic changes, rates of urbanisation and changes in dietary patterns are contributing to the changing trends in chronic disease in India. There is clear evidence of a demographic, epidemiological and nutrition transition in India that is fuelling the epidemic of chronic diseases and obesity, particularly in the urban areas.


Brain network reconfiguration for narrative and argumentative thought.
Our brain constructs reality through narrative and argumentative thought. Some hypotheses argue that these two modes of cognitive functioning are irreducible, reflecting distinct mental operations underlain by separate neural bases; Others ascribe both to a unitary neural system dedicated to long-timescale information. We addressed this question by employing inter-subject measures to investigate the stimulus-induced neural responses when participants were listening to narrative and argumentative texts during fMRI. We found that following both kinds of texts enhanced functional couplings within the frontoparietal control system. However, while a narrative specifically implicated the default mode system, an argument specifically induced synchronization between the intraparietal sulcus in the frontoparietal control system and multiple perisylvian areas in the language system. Our findings reconcile the two hypotheses by revealing commonalities and differences between the narrative and the argumentative brain networks, showing how diverse mental activities arise from the segregation and integration of the existing brain systems.


Effect of early neonatal vitamin A supplementation on mortality during infancy in Ghana (Neovita): a randomised, double-blind, placebo-controlled trial.
Results of randomised controlled trials of newborn (age 1-3 days) vitamin A supplementation have been inconclusive. The WHO is coordinating three large randomised trials in Ghana, India, and Tanzania (Neovita trials). We present the findings of the Neovita trial in Ghana. This study was a population-based, individually randomised, double-blind, placebo-controlled trial in the Brong Ahafo region of Ghana. The trial participants were infants aged at least 2 h, identified at home or facilities on the day of birth or in the next 2 days, able to feed orally, and likely to stay in the study area for at least 6 months. They were randomly assigned (ratio 1:1) to receive either one oral dose of vitamin A (50,000 IU) or placebo immediately after recruitment. The research team and parents of the infants were masked to treatment assignment. Follow-up home visits were undertaken every 4 weeks, when data were recorded for deaths, facility use, and care seeking. The primary outcome was post-supplementation mortality to 6 months of age. Analysis was by intention to treat. Potential adverse events were recorded at 1 and 3 days after supplementation. This trial is registered with the Australian New Zealand Clinical Trials Registry (ANZCTR)CTRN12610000582055. We assessed 26,414 livebirths for eligibility between Aug 16, 2010, and Nov 7, 2011. We recruited 22,955 newborn infants, with 11,474 randomly assigned to receive vitamin A and 11,481 to receive placebo. Loss to follow-up was low with vital status at 6 months of age reported for 22,698 (98·9%) infants. We recorded 278 post-supplementation deaths to 6 months of age in the vitamin A group (mortality risk 24·5 in 1000 supplemented infants) and 248 deaths in the placebo group (mortality risk 21·8 per 1000 supplemented infants), relative risk (RR) 1·12 (95% CI 0·95-1·33; p=0·183) and risk difference (RD) 2·66 (95% CI -1·25 to 6·57; p=0·18). Adverse events within 3 days of supplementation did not differ by trial group. 122 infants died in the first 3 days after supplementation; 70 (0·6%) in the vitamin A and 52 (0·5%) in the placebo group (risk ratio [RR] 1·35, 95% CI 0·94-1·93, p=0·102). 53 infants were reported to have a bulging fontanelle; 32 (0·3%) in the vitamin A group and 21 (0·2%) in the placebo group (RR 1·53, 0·88-2·62, p=0·130). The results of this trial do not support inclusion of newborn vitamin A supplementation as a child survival strategy in Ghana. Bill & Melinda Gates Foundation grant to the WHO.


Controlling Electronic Devices with Brain Rhythms/Electrical Activity Using Artificial Neural Network (ANN).
The purpose of this research study was to explore the possibility to develop a brain-computer interface (BCI). The main objective was that the BCI should be able to recognize brain activity. BCI is an emerging technology which focuses on communication between software and hardware and permitting the use of brain activity to control electronic devices, such as wheelchairs, computers and robots. The interface was developed, and consists of EEG Bitronics, Arduino and a computer; moreover, two versions of the BCIANNET software were developed to be used with this hardware. This BCI used artificial neural network (ANN) as a main processing method, with the Butterworth filter used as the data pre-processing algorithm for ANN. Twelve subjects were measured to collect the datasets. Tasks were given to subjects to stimulate brain activity. The purpose of the experiments was to test and confirm the performance of the developed software. The aim of the software was to separate important rhythms such as alpha, beta, gamma and delta from other EEG signals. As a result, this study showed that the Levenberg-Marquardt algorithm is the best compared with the backpropagation, resilient backpropagation, and error correction algorithms. The final developed version of the software is an effective tool for research in the field of BCI. The study showed that using the Levenberg-Marquardt learning algorithm gave an accuracy of prediction around 60% on the testing dataset.


Indoor secondary pollutants from household product emissions in the presence of ozone: A bench-scale chamber study.
Ozone-driven chemistry is a source of indoor secondary pollutants of potential health concern. This study investigates secondary air pollutants formed from reactions between constituents of household products and ozone. Gas-phase product emissions were introduced along with ozone at constant rates into a 198-L Teflon-lined reaction chamber. Gas-phase concentrations of reactive terpenoids and oxidation products were measured. Formaldehyde was a predominant oxidation byproduct for the three studied products, with yields for most conditions of 20-30% with respect to ozone consumed. Acetaldehyde, acetone, glycolaldehyde, formic acid, and acetic acid were each also detected for two or three of the products. Immediately upon mixing of reactants, a scanning mobility particle sizer detected particle nucleation events that were followed by a significant degree of secondary particle growth. The production of secondary gaseous pollutants and particles depended primarily on the ozone level and was influenced by other parameters such as the air-exchange rate. Hydroxyl radical concentrations in the range 0.04-200 x 10(5) molecules cm(-3) were determined by an indirect method. OH concentrations were observed to vary strongly with residual ozone level in the chamber, which was in the range 1-25 ppb, as is consistent with expectations from a simplified kinetic model. In a separate chamber study, we exposed the dry residue of two products to ozone and observed the formation of gas-phase and particle-phase secondary oxidation products.


Improving performance of HVAC systems to reduce exposure to aerosolized infectious agents in buildings; recommendations to reduce risks posed by biological attacks.
The prospect of biological attacks is a growing strategic threat. Covert aerosol attacks inside a building are of particular concern. In the summer of 2005, the Center for Biosecurity of the University of Pittsburgh Medical Center convened a Working Group to determine what steps could be taken to reduce the risk of exposure of building occupants after an aerosol release of a biological weapon. The Working Group was composed of subject matter experts in air filtration, building ventilation and pressurization, air conditioning and air distribution, biosecurity, building design and operation, building decontamination and restoration, economics, medicine, public health, and public policy. The group focused on functions of the heating, ventilation, and air conditioning systems in commercial or public buildings that could reduce the risk of exposure to deleterious aerosols following biological attacks. The Working Group's recommendations for building owners are based on the use of currently available, off-the-shelf technologies. These recommendations are modest in expense and could be implemented immediately. It is also the Working Group's judgment that the commitment and stewardship of a lead government agency is essential to secure the necessary financial and human resources and to plan and build a comprehensive, effective program to reduce exposure to aerosolized infectious agents in buildings.


The joint effect of asbestos exposure, tobacco smoking and alcohol drinking on laryngeal cancer risk: evidence from the French population-based case-control study, ICARE.
The objective of the study was to investigate the joint effect of occupational exposure to asbestos, and tobacco and alcohol consumption, on the risk of laryngeal cancer among men. We used data from a large population-based case-control study conducted in France. We estimated two-way and three-way interactions between asbestos exposure (never vs ever exposed), tobacco consumption (<20 vs. ≥20 pack-years) and alcohol consumption (<5 vs. ≥5 drinks per day). The interaction on an additive scale was assessed by estimating the relative excess risk due to interaction (RERI) and the attributable proportion due to interaction, and the interaction on a multiplicative scale was assessed by estimating the multiplicative interaction parameter (ψ). Multiplicative interactions were also assessed using fractional polynomials for alcohol drinking, tobacco smoking and asbestos exposure. When compared with light-to-moderate smokers and drinkers never exposed to asbestos, the increase in laryngeal cancer risk was smallest among light-to-moderate drinkers and smokers exposed to asbestos (OR=2.23 (1.08 to 4.60)), and highest among heavy smokers and drinkers ever exposed to asbestos (OR=69.39 (35.54 to 135.5)). We found an additive joint effect between asbestos exposure and alcohol consumption (RERI=4.75 (-4.29 to 11.12)), whereas we observed a more than additive joint effect between asbestos exposure and tobacco consumption (RERI=8.50 (0.71 to 23.81)), as well as between asbestos exposure, and tobacco and alcohol consumption (RERI=26.57 (11.52 to 67.88)). However, our results did not suggest any interaction on a multiplicative scale. Our results suggest that asbestos exposure, in combination with tobacco and alcohol exposure, accounted for a substantial number of laryngeal cancer cases. Our findings therefore highlight the need for prevention in activities, such as construction work, where exposure to asbestos-containing materials remains.


Striving for scientific stringency: a re-analysis of a randomised controlled trial considering first-time mothers' obstetric outcomes in relation to birth position.
The aim of this study was to compare maternal labour and birth outcomes between women who gave birth on a birth seat or in any other position for vaginal birth and further, to study the relationship between synthetic oxytocin augmentation and maternal blood loss, in a stratified sample. A re-analysis of a randomized controlled trial in Sweden. An on-treatment analysis was used to study obstetrical outcomes for nulliparous women who gave birth on a birth seat (birth seat group) compared to birth in any other position for vaginal birth (control group). Data were collected between November 2006 and July 2009. The outcome measurements included perineal outcome, post partum blood loss, epidural analgesia, synthetic oxytocin augmentation and duration of labour. The major findings of this paper were that women giving birth on the birth seat had shorter duration of labour and were significantly less likely to receive synthetic oxytocin for augmentation in the second stage of labour. Significantly more women had an increased blood loss when giving birth on the birth seat, but had no difference in perineal outcomes. Blood loss was increased regardless of birth position if women had been exposed to synthetic oxytocin augmentation during the first stage of labour. The results of this analysis imply that women with a straightforward birth process may well benefit from giving birth on a birth seat without risk for any adverse obstetrical outcomes. However it is important to bear in mind that, women who received synthetic oxytocin during the first stage of labour may have an increased risk for greater blood loss when giving birth on a birth seat. Finally it is of vital importance to scrutinize the influence of synthetic oxytocin administered during the first stage of labour on blood loss postpartum, since excessive blood loss is a well-documented cause of maternal mortality worldwide and may cause severe maternal morbidity in high-income countries. Unique Protocol ID: NCT01182038 ( http://register.clinicaltrials.gov).


Fibroblasts as immune regulators in infection, inflammation and cancer.
In chronic infection, inflammation and cancer, the tissue microenvironment controls how local immune cells behave, with tissue-resident fibroblasts emerging as a key cell type in regulating activation or suppression of an immune response. Fibroblasts are heterogeneous cells, encompassing functionally distinct populations, the phenotypes of which differ according to their tissue of origin and type of inciting disease. Their immunological properties are also diverse, ranging from the maintenance of a potent inflammatory environment in chronic inflammation to promoting immunosuppression in malignancy, and encapsulating and incarcerating infectious agents within tissues. In this Review, we compare the mechanisms by which fibroblasts control local immune responses, as well as the factors regulating their inflammatory and suppressive profiles, in different tissues and pathological settings. This cross-disease perspective highlights the importance of tissue context in determining fibroblast-immune cell interactions, as well as potential therapeutic avenues to exploit this knowledge for the benefit of patients with chronic infection, inflammation and cancer.


Aberrant transforming growth factor beta1 signaling and SMAD4 nuclear translocation confer epigenetic repression of ADAM19 in ovarian cancer.
Transforming growth factor-beta (TGF-beta)/SMAD signaling is a key growth regulatory pathway often dysregulated in ovarian cancer and other malignancies. Although loss of TGF-beta-mediated growth inhibition has been shown to contribute to aberrant cell behavior, the epigenetic consequence(s) of impaired TGF-beta/SMAD signaling on target genes is not well established. In this study, we show that TGF-beta1 causes growth inhibition of normal ovarian surface epithelial cells, induction of nuclear translocation SMAD4, and up-regulation of ADAM19 (a disintegrin and metalloprotease domain 19), a newly identified TGF-beta1 target gene. Conversely, induction and nuclear translocation of SMAD4 were negligible in ovarian cancer cells refractory to TGF-beta1 stimulation, and ADAM19 expression was greatly reduced. Furthermore, in the TGF-beta1 refractory cells, an inactive chromatin environment, marked by repressive histone modifications (trimethyl-H3K27 and dimethyl-H3K9) and histone deacetylase, was associated with the ADAM19 promoter region. However, the CpG island found within the promoter and first exon of ADAM19 remained generally unmethylated. Although disrupted growth factor signaling has been linked to epigenetic gene silencing in cancer, this is the first evidence demonstrating that impaired TGF-beta1 signaling can result in the formation of a repressive chromatin state and epigenetic suppression of ADAM19. Given the emerging role of ADAMs family proteins in growth factor regulation in normal cells, we suggest that epigenetic dysregulation of ADAM19 may contribute to the neoplastic process in ovarian cancer.


Divergent clonal selection dominates medulloblastoma at recurrence.
The development of targeted anti-cancer therapies through the study of cancer genomes is intended to increase survival rates and decrease treatment-related toxicity. We treated a transposon-driven, functional genomic mouse model of medulloblastoma with 'humanized' in vivo therapy (microneurosurgical tumour resection followed by multi-fractionated, image-guided radiotherapy). Genetic events in recurrent murine medulloblastoma exhibit a very poor overlap with those in matched murine diagnostic samples (<5%). Whole-genome sequencing of 33 pairs of human diagnostic and post-therapy medulloblastomas demonstrated substantial genetic divergence of the dominant clone after therapy (<12% diagnostic events were retained at recurrence). In both mice and humans, the dominant clone at recurrence arose through clonal selection of a pre-existing minor clone present at diagnosis. Targeted therapy is unlikely to be effective in the absence of the target, therefore our results offer a simple, proximal, and remediable explanation for the failure of prior clinical trials of targeted therapy.


Parameter estimation in fluorescence recovery after photobleaching: quantitative analysis of protein binding reactions and diffusion.
Fluorescence recovery after photobleaching (FRAP) is a common experimental method for investigating rates of molecular redistribution in biological systems. Many mathematical models of FRAP have been developed, the purpose of which is usually the estimation of certain biological parameters such as the diffusivity and chemical reaction rates of a protein, this being accomplished by fitting the model to experimental data. In this article, we consider a two species reaction-diffusion FRAP model. Using asymptotic analysis, we derive new FRAP recovery curve approximation formulae, and formally re-derive existing ones. On the basis of these formulae, invoking the concept of Fisher information, we predict, in terms of biological and experimental parameters, sufficient conditions to ensure that the values all model parameters can be estimated from data. We verify our predictions with extensive computational simulations. We also use computational methods to investigate cases in which some or all biological parameters are theoretically inestimable. In these cases, we propose methods which can be used to extract the maximum possible amount of information from the FRAP data.


High Prevalence of Dehydration and Inadequate Nutritional Knowledge Among University and Club Level Athletes.
Although dehydration of ≥ 2% body weight (BW) loss significantly impairs endurance performance, dehydration remains prevalent among athletes and may be owing to a lack of knowledge in relation to fluid requirements. The aim of this study was to assess the hydration status of university/club level athletes (n = 430) from a range of sports/activities (army officer cadet training; bootcamp training; cycling; Gaelic Athletic Association camogie, football and hurling; golf; hockey; netball; rugby; running (sprinting and endurance); Shotokan karate and soccer) immediately before and after training/competition and to assess their nutritional knowledge. Urine specific gravity (USG) was measured immediately before and after exercise and BW loss during exercise was assessed. Nutritional knowledge was assessed using a validated questionnaire. 31.9% of athletes commenced exercise in a dehydrated state (USG >1.020) with 43.6% of participants dehydrated posttraining/competition. Dehydration was particularly prevalent (>40% of cohort) among karateka, female netball players, army officer cadets, and golfers. Golfers that commenced a competitive 18 hole round dehydrated took a significantly higher number of strokes to complete the round in comparison with their euhydrated counterparts (79.5 ± 2.1 vs. 75.7 ± 3.9 strokes, p = .049). Nutritional knowledge was poor among participants (median total score [IQR]; 52.9% [46.0, 59.8]), albeit athletes who were euhydrated at the start of exercise had a higher overall score in comparison with dehydrated athletes (55.2% vs. 50.6%, p = .001). Findings from the current study, therefore, have significant implications for the education of athletes in relation to their individual fluid requirements around exercise.


Opposing Effects of Granulocyte Colony-Stimulating Factor on the Initiation and Progression of Breast Cancer Bone Metastases.
Granulocyte colony stimulating factor (G-CSF), an essential cytokine regulating granulopoiesis, is expressed in a substantial proportion of breast cancers, and it has been implicated in cancer progression. Here, we examined effects of G-CSF on the development of bone metastases of breast cancer using immunocompetent mouse models. The expression of CXC chemokine ligand 12 (CXCL12) in bone marrow stromal cells, which plays a critical role in the maintenance of hematopoietic stem cells and also in cancer cell homing to bone, was markedly decreased in mice treated with G-CSF. Flow cytometric analysis revealed that pretreatment of mice with G-CSF reduced the number of bone-homing cancer cells. G-CSF also increased the population of myeloid-derived suppressor cells (MDSCs) in bone marrow. Depletion of MDSCs using anti-Gr-1 antibody treatment significantly decreased the metastatic tumor burden in bone. The overall effects of G-CSF on bone metastases were finally examined using two different treatment protocols. When mice were treated with G-CSF prior to the tumor cell inoculation, G-CSF did not change bone metastatic-tumor burden. In contrast, when G-CSF treatment was started after the tumor cells had homed to bone, G-CSF significantly accelerated bone metastases formation. These results suggest that G-CSF suppressed cancer cell homing to bone by downregulating CXCL12 expression in bone marrow stromal cells, whereas G-CSF stimulated the progression of bone metastases at least in part by MDSC-mediated mechanisms. IMPLICATIONS: G-CSF had opposing effects on the initiation and progression of bone metastases of breast cancer and the balance may regulate the metastatic tumor burden.


Shifting priorities: highly conserved behavioral and brain network adaptations to chronic stress across species.
Parallel clinical and preclinical research have begun to illuminate the biological basis of stress-related disorders, including major depression, but translational bridges informing discrete mechanistic targets for intervention are missing. To address this critical need, we used structural MRI in a mouse model and in a large human sample to examine stress effects on brain structure that may be conserved across species. Specifically, we focused on a previously unexplored approach, whole-brain structural covariance, as it reflects synchronized changes in neuroanatomy, potentially due to mutual trophic influences or shared plasticity across regions. Using the unpredictable chronic mild stress (UCMS) paradigm in mouse we first demonstrate that UCMS-induced elevated behavioral emotionality correlates with increased size of the amygdala and other corticolimbic regions. We further identify focal increases in the amygdala's 'hubness' (degree and strength) set against the background of a global stress-related loss of network clustering and modularity. These macroscopic changes are supported on the molecular level by increased postsynaptic density-95 protein in the amygdala, consistent with stress-induced plastic changes and synaptic strengthening. Finally, we provide clinical evidence that strikingly similar structural network reorganization patterns exist in young adults reporting high childhood trauma and increased mood symptoms. Collectively, we provide initial translational evidence for a conserved stress-related increase in amygdala-centered structural synchrony, as measured by enhanced structural covariance, which is paralleled by a decrease in global structural synchrony. This putative trade-off reflected in increased amygdala-centered plastic changes at the expense of global structural dedifferentiation may represent a mechanistic pathway for depression and related psychopathology.


The Instrumented Sit-to-Stand Test (iSTS) Has Greater Clinical Relevance than the Manually Recorded Sit-to-Stand Test in Older Adults.
The ability to rise from sitting to standing is critical to an individual's quality of life, as it is a prerequisite for functional independence. The purpose of the current study was to examine the hypothesis that test durations as assessed with the instrumented repeated Sit-To-Stand (STS) show stronger associations with health status, functional status and daily physical activity of older adults than manually recorded test durations. In 63 older participants (mean age 83 ±6.9 years, 51 female), health status was assessed using the European Quality of Life questionnaire and functional status was assessed using the physical function index of the of the RAND-36. Physical performance was measured using a wearable sensor-based STS test. From this test, durations, sub-durations and kinematics of the STS movements were estimated and analysed. In addition, physical activity was measured for one week using an activity monitor and episodes of lying, sitting, standing and locomotion were identified. Associations between STS parameters with health status, functional status and daily physical activity were assessed. The manually recorded STS times were not significantly associated with health status (p = 0.457) and functional status (p = 0.055), whereas the instrumented STS times were (both p = 0.009). The manually recorded STS durations showed a significant association to daily physical activity for mean sitting durations (p = 0.042), but not for mean standing durations (p = 0.230) and mean number of locomotion periods (p = 0.218). Furthermore, durations of the dynamic sit-to-stand phase of the instrumented STS showed more significant associations with health status, functional status and daily physical activity (all p = 0.001) than the static phases standing and sitting (p = 0.043-0.422). As hypothesized, instrumented STS durations were more strongly associated with participant health status, functional status and physical activity than manually recorded STS durations in older adults. Furthermore, instrumented STS allowed assessment of the dynamic phases of the test, which were likely more informative than the static sitting and standing phases.


Knocking-down the NMDAR1 subunit in a limited amount of neurons in the rat hippocampus impairs learning.
Amplicon vectors derived from herpes simplex virus type 1 were built to modify NMDA receptors by expressing antisense RNA for the essential NR1 subunit. Their ability to modify endogenous levels of NR1 was tested in cultures of rat embryo neocortical neurons. We studied behaviour and tested for expression in adult rats injected with those vectors into the dorsal hippocampus to find out which cells and how many appear involved in memory formation. Rats injected with vectors expressing NR1 antisense performed significantly worse than control rats in an inhibitory avoidance task. Immunohistochemistry was performed in brain slices from the same animals. The transduced cells represented 6-7% of pyramidal neurons in CA1, showing that a single gene knockdown of NR1 in a small number of neurons significantly impaired memory formation. Perhaps neurons undergoing synaptic plasticity are more susceptible to NR1 knockdown, and hence NMDAR are particularly required in those neurons undergoing synaptic plasticity during learning, or perhaps, and more likely, there is not a high level of redundancy in the hippocampal circuits involved, leading to the idea that a certain level of NR1 expression/availability appears necessary for memory formation in most of CA1 pyramidal neurons.


Characterization of tissue transglutaminase in human osteoblast-like cells.
Tissue transglutaminase (tTG) is a calcium-dependent and guanosine 5'-triphosphate (GTP) binding enzyme, which catalyzes the post-translational modification of proteins by forming intermolecular epsilon(gamma-glutamyl)lysine cross-links. In this study, human osteoblasts (HOBs) isolated from femoral head trabecular bone and two osteosarcoma cell lines (HOS and MG-63) were studied for their expression and localization of tTG. Quantitative evaluation of transglutaminase (TG) activity determined using the [1,4 14C]-putrescine incorporation assay showed that the enzyme was active in all cell types. However, there was a significantly higher activity in the cell homogenates of MG-63 cells as compared with HOB and HOS cells (p < 0.001). There was no significant difference between the activity of the enzyme in HOB and HOS cells. All three cell types also have a small amount of active TG on their surface as determined by the incorporation of biotinylated cadaverine into fibronectin. Cell surface-related tTG was further shown by preincubation of cells with tTG antibody, which led to inhibition of cell attachment. Western blot analysis clearly indicated that the active TG was tTG and immunocytochemistry showed it be situated in the cytosol of the cells. In situ extracellular enzyme activity also was shown by the cell-mediated incorporation of fluorescein cadaverine into extracellular matrix (ECM) proteins. These results clearly showed that MG-63 cells have high extracellular activity, which colocalized with the ECM protein fibronectin and could be inhibited by the competitive primary amine substrate putrescine. The contribution of tTG to cell surface/matrix interactions and to the stabilization of the ECM of osteoblast cells therefore could by an important factor in the cascade of events leading to bone differentiation and mineralization.


The RNA editing enzyme APOBEC1 induces somatic mutations and a compatible mutational signature is present in esophageal adenocarcinomas.
The AID/APOBECs are deaminases that act on cytosines in a diverse set of pathways and some of them have been linked to the onset of genetic alterations in cancer. Among them, APOBEC1 is the only family member to physiologically target RNA, as the catalytic subunit in the Apolipoprotein B mRNA editing complex. APOBEC1 has been linked to cancer development in mice but its oncogenic mechanisms are not yet well understood. We analyze whether expression of APOBEC1 induces a mutator phenotype in vertebrate cells, likely through direct targeting of genomic DNA. We show its ability to increase the inactivation of a stably inserted reporter gene in a chicken cell line that lacks any other AID/APOBEC proteins, and to increase the number of imatinib-resistant clones in a human cellular model for chronic myeloid leukemia through induction of mutations in the BCR-ABL1 fusion gene. Moreover, we find the presence of an AID/APOBEC mutational signature in esophageal adenocarcinomas, a type of tumor where APOBEC1 is expressed, that mimics the one preferred by APOBEC1 in vitro. Our findings suggest that the ability of APOBEC1 to trigger genetic alterations represents a major layer in its oncogenic potential. Such APOBEC1-induced mutator phenotypes could play a role in the onset of esophageal adenocarcinomas. APOBEC1 could be involved in cancer promotion at the very early stages of carcinogenesis, as it is highly expressed in Barrett's esophagus, a condition often associated with esophageal adenocarcinoma.


Higher usual alcohol consumption was associated with a lower 41-y mortality risk from coronary artery disease in men independent of genetic and common environmental factors: the prospective NHLBI Twin Study.
Evidence that alcohol consumption is inversely associated with long-term coronary artery disease (CAD) mortality independent of genetic and early life environmental factors is lacking. We evaluated whether alcohol consumption was prospectively associated with CAD mortality risk independent of familial factors. In total, 843 male twins (396 pairs and 51 unpaired twins) aged 42-55 y (mean: 48 y) without baseline CAD reported beer, wine, and spirits consumption at baseline (1969-1973) and were followed up to 2010 in the prospective National Heart, Lung, and Blood Institute Twin Study. Data on usual alcohol consumption over the past year were collected. Outcome was time to event, where the primary event was death from CAD and secondary events were death from cardiovascular disease and all causes. HRs were estimated by using frailty survival models, both overall and within-pair. There were 129 CAD deaths and 219 cardiovascular deaths during 41 y of follow-up. In the whole cohort, after adjustment for caloric intake and cardiovascular disease risk factors, overall HRs per 10-g increment in alcohol intake were 0.94 (95% CI: 0.89, 0.98) for CAD and 0.97 (95% CI: 0.93, 1.00) for cardiovascular mortality. The within-pair adjusted HRs for a twin with 10-g higher daily alcohol consumption than his co-twin were 0.90 (95% CI: 0.84, 0.97) for CAD and 0.95 (95% CI: 0.90, 1.00) for cardiovascular disease mortality in the cohort pooled by zygosity, which remained similar among monozygotic twins. All 3 beverage types tended to be associated with lower CAD mortality risk within-pair to a similar degree. Alcohol consumption was not associated with total mortality risk overall or within-pair. Higher usual alcohol consumption is associated with lower CAD mortality risk, independent of germline and early life environment and adulthood experience shared among twins, supporting a possible causal role of alcohol consumption in lowering CAD death risk. This trial was registered at clinicaltrials.gov as NCT00005124.


Prediction of bulk tank somatic cell count violations based on monthly individual cow somatic cell count data.
The regulatory limit in Canada for bulk tank somatic cell count (BTSCC) was recently lowered from 500,000 to 400,000 cells/mL. Herd indices based on changes in cow somatic cell count over 2 consecutive months (e.g., proportion of healthy or chronically infected cows, cows cured, and new intramammary infection rate) could be used as predictors for BTSCC violations. The objective of this study was to develop a predictive model for exceeding the limit of 400,000 cells/mL in the next month using these herd indices. Dairy Herd Improvement (DHI) data were used from 924 dairy herds in Québec, Canada. Test-day BTSCC was estimated by dividing the sum of all cows' DHI test-day somatic cell count times DHI test-day milk production by the total volume of milk produced by the herd on that test-day. In total, 986 of 8,681 (11.4%) estimated BTSCC exceeded 400,000 cells/mL. The final predictive model included 6 variables: mean herd somatic cell score at the current test-month, proportion of cows >500,000 cells/mL at the current test-month, proportion of healthy cows during lactation at the current test-month, proportion of chronically infected cows at the current test-month, average days in milk at the current test-month, and annual mean daily milk production. The optimized sensitivity and specificity of the model were 76 and 74%, respectively. The positive predictive value and negative predictive value were 25 and 95%, respectively. This low positive predictive value and high negative predictive value demonstrated that the model was less accurate at predicting herds that would violate the estimated BTSCC threshold but very accurate at identifying herds that would not. In addition, the area under the curve for the receiver operating characteristic curve was 0.82, suggesting that the model had excellent discrimination between test-months that did and did not exceed 400,000 cells/mL. An internal validation was completed using a bootstrapped resampling-based estimation method and confirmed that the final model provided a validated estimate of predictive accuracy. This model could be used to monitor and advise clients on impending risks of exceeding the BTSCC limit.


A year in review in Minerva Anestesiologica 2013.
We organized a training program for oral fiber optic intubation (FOI) under conscious sedation. The efficacy of the program was evaluated by comparing the performances of experts and novices. The training procedure was divided into two sessions: a theoretical session on difficult airways, the fiber optic bronchoscope (FOB), remifentanil, topical anesthesia and patient interactions; and a session involving simulations of the FOI technique on dummies. For in vivo FOI, we enrolled patients requiring orotracheal intubation for elective surgery. Electrocardiograms, mean arterial pressure was railroaded over the fiberscope, and tracheal intubati6 and 7) FOIs, respectively, joined the study. To reach ±23 bpm, P=0.02), and RR was decreased (from 16±3 to 12±4 bpm, P<0.05). No differences were recorded between the experts and less-experienced anesthesiologists. The average duration of FOI was 3.3±2.0 min for experts and 4.2±2.4 min for novices (P=0.03). Procedures were successful in both groups, with patients in each group being equally satisfied with the procedures. This study highlights the importance of a structured FOI training program, demonstrating that it is possible to learn to perform FOI proficiently by practicing on dummies.


Mathematically gifted adolescents have deficiencies in social valuation and mentalization.
Many mathematically gifted adolescents are characterized as being indolent, underachieving and unsuccessful despite their high cognitive ability. This is often due to difficulties with social and emotional development. However, research on social and emotional interactions in gifted adolescents has been limited. The purpose of this study was to observe differences in complex social strategic behaviors between gifted and average adolescents of the same age using the repeated Ultimatum Game. Twenty-two gifted adolescents and 24 average adolescents participated in the Ultimatum Game. Two adolescents participate in the game, one as a proposer and the other as a responder. Because of its simplicity, the Ultimatum Game is an apt tool for investigating complex human emotional and cognitive decision-making in an empirical setting. We observed strategic but socially impaired offers from gifted proposers and lower acceptance rates from gifted responders, resulting in lower total earnings in the Ultimatum Game. Thus, our results indicate that mathematically gifted adolescents have deficiencies in social valuation and mentalization.


GABAergic antagonists block the inhibitory effects of serotonin in the lateral amygdala: a mechanism for modulation of sensory inputs related to fear conditioning.
Neurons in the lateral amygdala (LA) receive glutamatergic sensory input from the auditory thalamus and auditory cortex, and these inputs can be modulated by serotonin (5-HT). In the present study, we examined whether serotonergic inhibition of glutamatatergic excitation in the LA occurs via activation of GABAergic interneurons. Single-unit extracellular activity in the LA was recorded in response to iontophoretically applied glutamate. Concurrent application of 5-HT reduced the number of glutamate-evoked action potentials in the majority of neurons tested. GABA antagonists were then iontophoresed with both glutamate and 5-HT. Of the neurons that were inhibited by 5-HT, concurrent application of the GABA antagonists significantly reversed this effect. Application of the GABA antagonists alone had little or no effect on basal neuronal activity. We conclude that the 5-HT-induced inhibition of glutamatergic activity occurs in part through activation of serotonergic receptors on GABAergic interneurons.


Multiple paleopolyploidizations during the evolution of the Compositae reveal parallel patterns of duplicate gene retention after millions of years.
Of the approximately 250,000 species of flowering plants, nearly one in ten are members of the Compositae (Asteraceae), a diverse family found in almost every habitat on all continents except Antarctica. With an origin in the mid Eocene, the Compositae is also a relatively young family with remarkable diversifications during the last 40 My. Previous cytologic and systematic investigations suggested that paleopolyploidy may have occurred in at least one Compositae lineage, but a recent analysis of genomic data was equivocal. We tested for evidence of paleopolyploidy in the evolutionary history of the family using recently available expressed sequence tag (EST) data from the Compositae Genome Project. Combined with data available on GenBank, we analyzed nearly 1 million ESTs from 18 species representing seven genera and four tribes. Our analyses revealed at least three ancient whole-genome duplications in the Compositae-a paleopolyploidization shared by all analyzed taxa and placed near the origin of the family just prior to the rapid radiation of its tribes and independent genome duplications near the base of the tribes Mutisieae and Heliantheae. These results are consistent with previous research implicating paleopolyploidy in the evolution and diversification of the Heliantheae. Further, we observed parallel retention of duplicate genes from the basal Compositae genome duplication across all tribes, despite divergence times of 33-38 My among these lineages. This pattern of retention was also repeated for the paleologs from the Heliantheae duplication. Intriguingly, the categories of genes retained in duplicate were substantially different from those in Arabidopsis. In particular, we found that genes annotated to structural components or cellular organization Gene Ontology categories were significantly enriched among paleologs, whereas genes associated with transcription and other regulatory functions were significantly underrepresented. Our results suggest that paleopolyploidy can yield strikingly consistent signatures of gene retention in plant genomes despite extensive lineage radiations and recurrent genome duplications but that these patterns vary substantially among higher taxonomic categories.


Biomechanical evaluation of subcrestal dental implants with different bone anchorages.
This study evaluated the biomechanical influence of apical bone anchorage on a single subcrestal dental implant using three-dimensional finite element analysis (FEA). Four different bone anchorage designs were simulated on a posterior maxillary segment using one implant with platform switching and internal Morse taper connection as follows: 2 mm subcrestal placement with (SW) or without (SO) the implant apex engaged into the cortical bone or position at bone level with anchorage only in the crestal cortical (BO) bone or with bicortical fixation (BW). Each implant received a premolar crown, and all models were loaded with 200 N to simulate centric and eccentric occlusion. The peak tensile and compressive stress and strain were calculated at the crestal cortical, trabecular, and apical cortical bone. The vertical and horizontal implant displacements were measured at the platform level. FEA indicated that subcrestal placement (SW and SO) created lower stress and strain in the crestal cortical bone compared with crestal placement (BO and BW models). The SW model exhibited lesser vertical and horizontal implant micromovement compared with the SO and BO models under eccentric loading; however, stress and strain were higher in the apical cortical bone. The BW model exhibited the lowest implant displacement. These results indicate that subcrestal placement decreases the stress in the crestal cortical bone of dental implants, regardless of apical anchorage; however, apical cortical anchorage can be effective in limiting implant displacement. Further studies are required to evaluate the effects of possible remodeling around the apex on the success of subcrestal implants.


Re-irradiation and external hyperthermia in locally advanced, radiation recurrent, hormone refractory prostate cancer: a preliminary report.
The purpose of this report is to present the preliminary results of re-irradiation and external hyperthermia in patients with locally advanced, previously irradiated, hormone refractory prostate cancer. Three consecutive patients with symptomatic, locally advanced, previously irradiated and hormone refractory prostate cancer were treated with further irradiation (30.6-50 Gy) and external hyperthermia (5-8 treatments). All patients had complete resolution of symptoms lasting for 12-24 months. Significant tumour shrinkage, including complete tumour response, was demonstrated by CT and endoscopy. In one case, at 2 years after re-treatment, there is continued tumour regression and bone regeneration in the pelvis. Two patients had local control of tumour, which continued until most recent follow-up at 12 months and more than 24 months, respectively. Another case developed local recurrence at 17 months. At most recent follow-up, no patient has experienced significant treatment-related side effects. In these patients with no other therapeutic alternatives, re-irradiation and hyperthermia can provide durable tumour response for more than a year, resulting in significant improvement in quality of life. Further clinical studies are warranted.


The Association Between Trabecular Bone Score and Lumbar Spine Volumetric BMD Is Attenuated Among Older Men With High Body Mass Index.
Trabecular bone score (TBS) has been proposed as a dual-energy X-ray absorptiometry (DXA) derived measure of underlying quality of trabecular bone; however, TBS is not considered valid for those with body mass index (BMI) >37 kg/m2 . Our objective was to determine the association between TBS and lumbar spine (trabecular) volumetric BMD (LS-VBMD) and to examine whether the association varied by BMI and body composition among older men below this clinical threshold. We used regression models to study 3479 men age ≥65 years enrolled in the Osteoporotic Fractures in Men (MrOS) study who had TBS from spine DXA scans, LS-VBMD from central quantitative computed tomography, measures of trunk fat and lean mass from DXA, and BMI <37 kg/m2 . TBS was categorized as normal (n = 925), partially degraded (n = 1747), and degraded (n = 807). TBS was inversely related to BMI, trunk fat mass, and trunk lean mass (all p < 0.001). The relationship between TBS and LS-VBMD was nonlinear with magnitude of effect (slope of regression line using standardized variables) ranging from 0.07 (95% CI, -0.02 to 0.15) among those with degraded TBS up to 0.71 (95% CI, 0.54 to 0.89) among those with normal TBS. The relationship was still nonlinear after adjusting for age, clinical site, and either BMI, trunk lean mass, or trunk fat mass. The magnitude of effect relating TBS and LS-VBMD also decreased with increasing BMI (interaction, p = 0.090) and increasing trunk lean mass (interaction, p = 0.001), but not with increasing trunk fat mass (interaction, p = 0.224). In summary, the strength of the association between TBS and LS-VBMD among older men was variable and dependent on BMI and body composition, particularly trunk lean mass. The clinical utility of TBS among older men may be somewhat limited among men with high BMI or high trunk lean mass. © 2016 American Society for Bone and Mineral Research.


Beyond energies: geometry predictions with the XYG3 type of doubly hybrid density functionals.
The exchange-correlation functional holds the key to the success or failure of density functional theory calculations. When evaluating the functional performances, the current literature has focused much more on the ability of the method to reproduce exact electronic energies than on exact geometries. As all calculations have to start from the right geometries, the present feature article explores the functional performances on geometry predictions involving covalently and non-covalently bonded systems, as well as transition state structures with emphasis on the XYG3 type of doubly hybrid (xDH) functionals. The results are also presented for some challenging cases, demonstrating the usefulness of the xDH functionals in general.


Ratiometric analysis of fura red by flow cytometry: a technique for monitoring intracellular calcium flux in primary cell subsets.
Calcium flux is a rapid and sensitive measure of cell activation whose utility could be enhanced with better techniques for data extraction. We describe a technique to monitor calcium flux by flow cytometry, measuring Fura Red calcium dye by ratiometric analysis. This technique has several advantages: 1) using a single calcium dye provides an additional channel for surface marker characterization, 2) allows robust detection of calcium flux by minority cell populations within a heterogeneous population of primary T cells and monocytes 3) can measure total calcium flux and additionally, the proportion of responding cells, 4) can be applied to studying the effects of drug treatment, simultaneously stimulating and monitoring untreated and drug treated cells. Using chemokine receptor activation as an example, we highlight the utility of this assay, demonstrating that only cells expressing a specific chemokine receptor are activated by cognate chemokine ligand. Furthermore, we describe a technique for simultaneously stimulating and monitoring calcium flux in vehicle and drug treated cells, demonstrating the effects of the Gαi inhibitor, pertussis toxin (PTX), on chemokine stimulated calcium flux. The described real time calcium flux assay provides a robust platform for characterizing cell activation within primary cells, and offers a more accurate technique for studying the effect of drug treatment on receptor activation in a heterogeneous population of primary cells.


Study of interlayer spacing collapse during polymer/clay nanocomposite melt intercalation.
The influence of the chemical structure of alkylammonium organo-modifying montmorillonite clays on the ability to form nanocomposites by melt blending, depending on the processing temperature and the organoclay thermal treatment, has been investigated. On one side chlorinated polyethylene/Cloisite 30B (nano)composite has been prepared by melt intercalation at 175 degrees C and its wide angle X-ray diffraction pattern revealed that the peak characteristic of the interlayer spacing of the organoclay was shifted to lower d-spacing, indicating a collapse of the organoclay structure. On the other side, (nano)composites based on ethylene-vinyl acetate copolymer/Cloisite 30B have been prepared by melt intercalation at 140 degrees C. At this temperature, exfoliation was observed with the as-received organoclay while the same organo-modified clay, simply dried at 180 degrees C for 2 hours, induced again the formation of a composite with a collapsed structure. The effect of the Cloisite 30B thermal treatment on the morphology and mechanical properties of ethylene-vinyl acetate-based (nano)composites was investigated using wide angle X-ray diffraction and tensile tests. In order to shed some light on the origin of this clay interlayer collapse, organoclay modified with various ammonium cations bearing long alkyl chains with different amounts of unsaturations were studied using wide angle X-ray diffraction and X-ray photoelectron spectroscopy before and after thermal treatment at 180 degrees C for 2 hours. Isothermal thermogravimetric analysis of all organoclays was also investigated. The layers collapse effect is discussed depending upon the level of unsatured hydrocarbon present in the organic surfactant.


Identification of Intracellular and Extracellular Metabolites in Cancer Cells Using 13C Hyperpolarized Ultrafast Laplace NMR.
Ultrafast Laplace NMR (UF-LNMR), which is based on the spatial encoding of multidimensional data, enables one to carry out 2D relaxation and diffusion measurements in a single scan. Besides reducing the experiment time to a fraction, it significantly facilitates the use of nuclear spin hyperpolarization to boost experimental sensitivity, because the time-consuming polarization step does not need to be repeated. Here we demonstrate the usability of hyperpolarized UF-LNMR in the context of cell metabolism, by investigating the conversion of pyruvate to lactate in the cultures of mouse 4T1 cancer cells. We show that 13C ultrafast diffusion- T2 relaxation correlation measurements, with the sensitivity enhanced by several orders of magnitude by dissolution dynamic nuclear polarization (D-DNP), allows the determination of the extra- vs intracellular location of metabolites because of their significantly different values of diffusion coefficients and T2 relaxation times. Under the current conditions, pyruvate was located predominantly in the extracellular pool, while lactate remained primarily intracellular. Contrary to the small flip angle diffusion methods reported in the literature, the UF-LNMR method does not require several scans with varying gradient strength, and it provides a combined diffusion and T2 contrast. Furthermore, the ultrafast concept can be extended to various other multidimensional LNMR experiments, which will provide detailed information about the dynamics and exchange processes of cell metabolites.


The sodium channel {beta}3-subunit induces multiphasic gating in NaV1.3 and affects fast inactivation via distinct intracellular regions.
Electrical excitability in neurons depends on the activity of membrane-bound voltage gated sodium channels (Na(v)) that are assembled from an ion conducting α-subunit and often auxiliary β-subunits. The α-subunit isoform Na(v)1.3 occurs in peripheral neurons together with the Na(v) β3-subunit, both of which are coordinately up-regulated in rat dorsal root ganglion neurons after nerve injury. Here we examine the effect of the β3-subunit on the gating behavior of Na(v)1.3 using whole cell patch clamp electrophysiology in HEK-293 cells. We show that β3 depolarizes the voltage sensitivity of Na(v)1.3 activation and inactivation and induces biphasic components of the inactivation curve. We detect both a fast and a novel slower component of inactivation, and we show that the β3-subunit increases the fraction of channels inactivating by the slower component. Using CD and NMR spectroscopy, we report the first structural analysis of the intracellular domain of any Na(v) β-subunit. We infer the presence of a region within the β3-subunit intracellular domain that has a propensity to form a short amphipathic α-helix followed by a structurally disordered sequence, and we demonstrate a role for both of these regions in the selective stabilization of fast inactivation. The complex gating behavior induced by β3 may contribute to the known hyperexcitability of peripheral neurons under those physiological conditions where expression of β3 and Na(v)1.3 are both enhanced.


Not all stress is created equal: Acute, not ambient stress, impairs learning in high schizotypes.
Learning from feedback is essential for daily functioning, with factors that impact learning having implications for healthy and clinical populations. Reinforcement learning appears impaired across the psychosis continuum, with deficits reported in patients with psychotic disorders as well as high schizotypes from the general population. Stress can impair learning, and sensitivity to stress is present along the psychosis continuum. The aim of the present study was to understand if stress impairs reinforcement learning in those at the lower end of the psychosis continuum. We investigated both naturalistic stress in everyday life using daily hassles (Study 1: n = 70; 31% male, M age = 22.67 years) and acute psychosocial stress using the Trier Social Stress Test (Study 2: n = 57; 32% male, M age = 22.43 years). In the presence of naturalistic stress, learning did not differ across schizotypes. However, under acute psychosocial stress, high schizotypes experienced impaired learning. Our results suggest trail-and-error learning is robust to the ebbs and flows of everyday stress for high schizotypes; however, acute stress is associated with decrements in learning. This indicates that the magnitude of stressors should be considered when designing cognitive and functional interventions for those along the psychosis continuum.


Illuminating the lineage-specific diversification of resin glycoside acylsugars in the morning glory (Convolvulaceae) family using computational metabolomics.
Acylsugars are a class of plant defense compounds produced across many distantly related families. Members of the horticulturally important morning glory (Convolvulaceae) family produce a diverse sub-class of acylsugars called resin glycosides (RGs), which comprise oligosaccharide cores, hydroxyacyl chain(s), and decorating aliphatic and aromatic acyl chains. While many RG structures are characterized, the extent of structural diversity of this class in different genera and species is not known. In this study, we asked whether there has been lineage-specific diversification of RG structures in different Convolvulaceae species that may suggest diversification of the underlying biosynthetic pathways. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was performed from root and leaf extracts of 26 species sampled in a phylogeny-guided manner. LC-MS/MS revealed thousands of peaks with signature RG fragmentation patterns with one species producing over 300 signals, mirroring the diversity in Solanaceae-type acylsugars. A novel RG from Dichondra argentea was characterized using Nuclear Magnetic Resonance spectroscopy, supporting previous observations of RGs with open hydroxyacyl chains instead of closed macrolactone ring structures. Substantial lineage-specific differentiation in utilization of sugars, hydroxyacyl chains, and decorating acyl chains was discovered, especially among Ipomoea and Convolvulus - the two largest genera in Convolvulaceae. Adopting a computational, knowledge-based strategy, we further developed a high-recall workflow that successfully explained ~72% of the MS/MS fragments, predicted the structural components of 11/13 previously characterized RGs, and partially annotated ~45% of the RGs. Overall, this study improves our understanding of phytochemical diversity and lays a foundation for characterizing the evolutionary mechanisms underlying RG diversification.


Comparison of various hemoglobin polyoxyethylene conjugate solutions as resuscitative fluids after hemorrhagic shock.
Previous research suggested that splanchnic hypoperfusion occurs after resuscitation with certain acellular hemoglobin solutions. We examined the influence of maltose content and oxygen affinity on resuscitation with various hemoglobin polyoxyethylene conjugate solutions after hemorrhage. Fifteen swine underwent hemorrhage and equal volume resuscitation with pyridoxalated hemoglobin polyoxyethylene conjugate containing 0% or 8% maltose, or low P50 conjugate, which also contained 8% maltose. Five control animals were monitored but not bled. Regional blood flow was determined by using radioactive microspheres, gastric mucosal perfusion was estimated with tonometry, and gut histopathology was evaluated. All hemoglobin solutions produced vasoconstriction, manifested by elevated mean systemic and pulmonary artery pressures without a significant decrease in cardiac index compared with the sham group. Resuscitation with maltose-containing solutions elevated arterial and regional PCO2 and depressed arterial pH and gastric pHi (p < 0.05 for all). Splanchnic and renal blood flows were reduced in the low P50 + 8% maltose group (p < 0.05 vs. sham and baseline for renal blood flow), possibly indicating greater regional vasoconstriction in this group. Ileal mucosal damage was more severe in the maltose-containing groups and correlated with decreased pHi. Vasoconstriction occurred in all groups but was more severe in the low P50 + 8% maltose group. Maltose-containing solutions caused respiratory acidosis, decreased pHi, and histologic evidence of mucosal injury. Pyridoxalated hemoglobin polyoxyethylene conjugate without maltose was a superior resuscitation solution in this swine model.