Non-human Primate Papillomaviruses Share Similar Evolutionary Histories and Niche Adaptation as the Human Counterparts.
Given high genetic diversity of papillomaviruses (PV) and complex scenario of virus-host interaction, the genetic basis underlying the mechanisms of HPV carcinogenicity is not well understood. In an effort to evaluate the origin and evolution of PV pathogenicity, we collected paired oral, perianal, and genital swabs from a wild macaque population. Of the 117 surveyed macaques, 88 (75.2%) were positive for PV DNA in one or more sites, mostly common from genital swabs, followed by oral and perianal sites. All putative macaque PV types phylogenetically clustered into the genera Alpha-, Beta-, and Gammapapillomavirus, with a strong phylogeny-tropism association as observed in HPVs. Using a Bayesian Markov Chain Monte Carlo framework, we demonstrated ancient intra-host divergence of primate PVs in which multiple ancestors had split and adapted to specific host ecosystems at least 41 million years ago, prior to the speciation events of primate host species. Following subsequent divergence and niche adaptation, distinct but phylogenetically related PV types were transmitted to similar host ecosystems by closely related host animals when host speciation occurred, which may explain in part the origin of carcinogenicity of HPV type 16 (HPV16) and Macaca fascicularis PV type 3 (MfPV3) that evolved from a most recent common ancestor containing the determinants for cervicovaginal colonization and cervical cancer. The findings identifying evolutionary and biological relatedness between human and non-human primate PVs lay a genetic foundation for research on parasite-host interactions and carcinogenic outcomes, which will prove useful in further study of viral pathogenesis and host specificity. To better understand the origin and evolution of PV carcinogenicity associated with cervical cancer, we applied a combination of phylogenetic and bioinformatic analyses to investigate the genetic diversity of macaque papillomaviruses, and estimate divergence times of human and non-human primate PVs. The majority of both human and non-human primate PVs cluster into α-, β-, and γ-PVs, sharing similar evolutionary histories and biological properties to each other. The strong phylogeny-tropism association of primate PVs indicates an important role of niche adaptation and virus-host codivergence shaping the diversity of viral genomics, host specificity, immune exposure, and pathogenic property. Understanding the evolution of the family Papilloamviridae in general and the primate papillomaviruses in specific in relevant to virus-host interactions should provide important implications for viral pathogenesis and disease prevention.


Frostbite in the prairies: a 12-year review.
A 12-year retrospective review of inpatient frostbite injury was undertaken in three tertiary hospitals serving a northern prairie population of 650,000. In the 125 patients identified, the mean age was 41 years, with a range of 3 to 90 years. Predisposing factors were alcohol consumption (46 percent), psychiatric illness (17 percent), vehicular trauma (19 percent), vehicular failure (15 percent), and drug use (4 percent). Profound hypothermia with a core temperature less than 32 degrees C was present in 12 percent. Anatomic distribution was 19 percent upper extremity, 47 percent lower extremity, 31 percent combined upper and lower extremity, and 3 percent facial or trunk only. Factors correlating with amputation of parts (p < 0.05) were duration of exposure, lack of proper attire, remote site of injury, presence of wound infection, and delay in seeking treatment. Prophylactic systemic antibiotics did not decrease the incidence of wound infection. Comparison with nine other civilian series revealed striking similarities in the patient populations.


Transition state geometry of driven chemical reactions on time-dependent double-well potentials.
Reaction rates across time-dependent barriers are difficult to define and difficult to obtain using standard transition state theory approaches because of the complexity of the geometry of the dividing surface separating reactants and products. Using perturbation theory (PT) or Lagrangian descriptors (LDs), we can obtain the transition state trajectory and the associated recrossing-free dividing surface. With the latter, we are able to determine the exact reactant population decay and the corresponding rates to benchmark the PT and LD approaches. Specifically, accurate rates are obtained from a local description regarding only direct barrier crossings and to those obtained from a stability analysis of the transition state trajectory. We find that these benchmarks agree with the PT and LD approaches for obtaining recrossing-free dividing surfaces. This result holds not only for the local dynamics in the vicinity of the barrier top, but also for the global dynamics of particles that are quenched at the reactant or product wells after their sojourn over the barrier region. The double-well structure of the potential allows for long-time dynamics related to collisions with the outside walls that lead to long-time returns in the low-friction regime. This additional global dynamics introduces slow-decay pathways that do not result from the local transition across the recrossing-free dividing surface associated with the transition state trajectory, but can be addressed if that structure is augmented by the population transfer of the long-time returns.


Solution-based nanosensors for in-field detection with the naked eye.
Nanomaterials are revolutionising analytical applications with low-cost tests that enable detecting a target molecule in a few steps and with the naked eye. With this approach, non-experts can perform analyses on-site and without utilising electronic readers. This is advantageous in point-of-care diagnostics, in-field measurements and analyses performed in resource-constrained settings. Here we review the main strategies adopted for detecting analytes with the naked eye and at the point of need using plasmonic nanosensors, catalytic nanoparticles and fluorescent nanomaterials. Examples of the detection of ions, glucose, small molecules, peptides and proteins with the nanosensors are explained in detail.


Visual adaptation and the cone flicker electroretinogram.
This study examined the hypothesis that changes in the response properties of the human cone ERG during light adaptation represent the recovery of cone system responsiveness toward a dark-adapted value after an initial decrease in responsiveness at adapting field onset. The electroretinographic (ERG) responses to 31.1 Hz flicker were obtained under both dark-adapted and light-adapted conditions for stimulus luminances ranging from -1.42(-)+0.82 log cd sec/m2. At low stimulus luminances, flicker ERG amplitudes were larger under dark-adapted than under light-adapted conditions, consistent with the hypothesis. However, at high stimulus luminances, flicker ERG amplitudes obtained under light-adapted conditions were approximately double those recorded from the dark-adapted eye. Therefore, the increase in cone ERG amplitude that occurs during light adaptation at high stimulus luminances does not represent a return toward a dark-adapted level but instead entails a substantial enhancement above the dark-adapted value, by a mechanism that is presently unidentified.


Histone H2A and H4 N-terminal tails are positioned by the MEP50 WD repeat protein for efficient methylation by the PRMT5 arginine methyltransferase.
The protein arginine methyltransferase PRMT5 is complexed with the WD repeat protein MEP50 (also known as Wdr77 or androgen coactivator p44) in vertebrates in a tetramer of heterodimers. MEP50 is hypothesized to be required for protein substrate recruitment to the catalytic domain of PRMT5. Here we demonstrate that the cross-dimer MEP50 is paired with its cognate PRMT5 molecule to promote histone methylation. We employed qualitative methylation assays and a novel ultrasensitive continuous assay to measure enzyme kinetics. We demonstrate that neither full-length human PRMT5 nor the Xenopus laevis PRMT5 catalytic domain has appreciable protein methyltransferase activity. We show that histones H4 and H3 bind PRMT5-MEP50 more efficiently compared with histone H2A(1-20) and H4(1-20) peptides. Histone binding is mediated through histone fold interactions as determined by competition experiments and by high density histone peptide array interaction studies. Nucleosomes are not a substrate for PRMT5-MEP50, consistent with the primary mode of interaction via the histone fold of H3-H4, obscured by DNA in the nucleosome. Mutation of a conserved arginine (Arg-42) on the MEP50 insertion loop impaired the PRMT5-MEP50 enzymatic efficiency by increasing its histone substrate Km, comparable with that of Caenorhabditis elegans PRMT5. We show that PRMT5-MEP50 prefers unmethylated substrates, consistent with a distributive model for dimethylation and suggesting discrete biological roles for mono- and dimethylarginine-modified proteins. We propose a model in which MEP50 and PRMT5 simultaneously engage the protein substrate, orienting its targeted arginine to the catalytic site.


Disruption of the serine/threonine kinase 9 gene causes severe X-linked infantile spasms and mental retardation.
X-linked West syndrome, also called "X-linked infantile spasms" (ISSX), is characterized by early-onset generalized seizures, hypsarrhythmia, and mental retardation. Recently, we have shown that the majority of the X-linked families with infantile spasms carry mutations in the aristaless-related homeobox gene (ARX), which maps to the Xp21.3-p22.1 interval, and that the clinical picture in these patients can vary from mild mental retardation to severe ISSX with additional neurological abnormalities. Here, we report a study of two severely affected female patients with apparently de novo balanced X;autosome translocations, both disrupting the serine-threonine kinase 9 (STK9) gene, which maps distal to ARX in the Xp22.3 region. We show that STK9 is subject to X-inactivation in normal female somatic cells and is functionally absent in the two patients, because of preferential inactivation of the normal X. Disruption of the same gene in two unrelated patients who have identical phenotypes (consisting of early-onset severe infantile spasms, profound global developmental arrest, hypsarrhythmia, and severe mental retardation) strongly suggests that lack of functional STK9 protein causes severe ISSX and that STK9 is a second X-chromosomal locus for this disorder.


Impact of the lipid bilayer on energy transfer kinetics in the photosynthetic protein LH2.
Photosynthetic purple bacteria convert solar energy to chemical energy with near unity quantum efficiency. The light-harvesting process begins with absorption of solar energy by an antenna protein called Light-Harvesting Complex 2 (LH2). Energy is subsequently transferred within LH2 and then through a network of additional light-harvesting proteins to a central location, termed the reaction center, where charge separation occurs. The energy transfer dynamics of LH2 are highly sensitive to intermolecular distances and relative organizations. As a result, minor structural perturbations can cause significant changes in these dynamics. Previous experiments have primarily been performed in two ways. One uses non-native samples where LH2 is solubilized in detergent, which can alter protein structure. The other uses complex membranes that contain multiple proteins within a large lipid area, which make it difficult to identify and distinguish perturbations caused by protein-protein interactions and lipid-protein interactions. Here, we introduce the use of the biochemical platform of model membrane discs to study the energy transfer dynamics of photosynthetic light-harvesting complexes in a near-native environment. We incorporate a single LH2 from Rhodobacter sphaeroides into membrane discs that provide a spectroscopically amenable sample in an environment more physiological than detergent but less complex than traditional membranes. This provides a simplified system to understand an individual protein and how the lipid-protein interaction affects energy transfer dynamics. We compare the energy transfer rates of detergent-solubilized LH2 with those of LH2 in membrane discs using transient absorption spectroscopy and transient absorption anisotropy. For one key energy transfer step in LH2, we observe a 30% enhancement of the rate for LH2 in membrane discs compared to that in detergent. Based on experimental results and theoretical modeling, we attribute this difference to tilting of the peripheral bacteriochlorophyll in the B800 band. These results highlight the importance of well-defined systems with near-native membrane conditions for physiologically-relevant measurements.


Soil organic carbon dynamics matching ecological equilibrium theory.
The persistence of soil organic carbon (SOC) has traditionally been explained as a combination of recalcitrance properties and stabilization processes, which lead to the formation of complex organic compounds. However, recent conceptual advances and experimental evidence challenge this view. Here, we test these conceptual advances using a dynamic equilibrium theory of SOC founded on classic ecological theory. We postulate that the persistence of SOC is an equilibrium point where SOC losses resulting from continuous decomposition and SOC gains due to SOC protection are balanced. We show that we can describe the temporal dynamics of SOC remarkably well (average and median R 2 = 0.75) in publicly available SOC time series from experiments that investigated the effects of agricultural practices in arable soils. The predictive power of our simplistic model is not meant to compete with that of current multi-pool SOC models or recent developments that include microbial loops. The simplicity of our analysis can, however, show how the conceptual distinction between the forces that control SOC loss and gain, and their equilibrium, can shed light on SOC dynamics. Specifically, our analysis shows that, regardless of specific mechanisms, the persistence of SOC will depend on the ultimate equilibrium between SOC gains and losses, which may depend on environmental (e.g. temperature) and ecological (e.g. spatially structured microbial activities) factors and the relative roles of these factors. Future experimental studies should quantify these roles to formulate a new generation of SOC dynamics model.


Endoscopic retrograde cholangiopancreatography in the management of biliary complications after orthotopic liver transplantation.
biliary complications are frequent after orthotopic liver transplantation and the management of these complications with endoscopic retrograde cholangiopancreatography (ERCP) is available. The aims of the study were to analyze the experience in the endoscopic management of biliary complications after liver transplantation in a third level center. Furthermore, the factors associated with higher rates of technical and clinical success were determined. this was an observational retrospective study of ERCPs performed in patients with biliary complications after liver transplantation between February 2012 and January 2017. The factors analyzed were: demographics, time between transplantation and ERCP, indications for ERCP, strategy of stenting (only plastic stents, only self-expandable metallic stents, plastic followed by metallic stents and metallic followed by plastic stents), technical and clinical success and complications. one hundred and sixty-eight endoscopies were performed in 58 patients. Thirty-three patients (56.9%) presented with early complications. The most frequent indication for ERCP was anastomotic stenosis (57.8%). Technical success in the first ERCP was achieved in 43 patients (74.1%). Early onset of biliary complications was associated with higher rates of technical success (OR: 6.49; p: 0.036). Clinical success was obtained in 36 cases (62.1%). Patients with early complications had a higher probability of having good clinical response (OR: 11.16; p: 0.033). The results were worse in patients with only plastic stents (50% of clinical success). Eleven complications were observed among 168 ERCPs (6.54%), including two pancreatitis, five bleeding events, three cholangitis and one micro-perforation. ERCP is safe and useful in the management of biliary complications after liver transplantation. Early onset of complications is associated with better results. Some patients will need repeated procedures to obtain a good clinical response.


Overview of abortion cases with severe maternal outcomes in the WHO Multicountry Survey on Maternal and Newborn Health: a descriptive analysis.
To summarise individual and institutional characteristics of abortion-related severe maternal outcomes reported at health facilities. Secondary analysis of data from the WHO Multicountry Survey on Maternal and Newborn Health. 85 health facilities in 23 countries. 322 women with abortion-related severe maternal outcomes. Frequency distributions and comparisons of differences in characteristics between cases of maternal near miss and death using Fisher's exact tests of association. Individual and institutional characteristics and frequencies of potentially life-threatening conditions, and interventions provided to women with severe maternal outcomes, maternal near miss, and maternal death. Most women with abortion-related severe maternal outcomes (SMOs) were 20-34 years old (65.2%), married or cohabitating (92.3%), parous (84.2%), and presented with abortions resulting from pregnancies at less than 14 weeks of gestation (67.1%). The women who died were younger, more frequently without a partner, and had abortions at ≥14 weeks of gestation, compared with women with maternal near miss (MNM). Curettage was the most common mode of uterine evacuation. The provision of blood products and therapeutic antibiotics were the most common other interventions recorded for all women with abortion-related SMOs; those who died more frequently had antibiotics, laparotomy, and hysterectomy, compared with women with MNM. Although haemorrhage was the most common cause of abortion-related SMO, infection (alone and in combination with haemorrhage) was the most common cause of death. This analysis affirms a number of previously observed characteristics of women with abortion-related severe morbidity and mortality, despite the fact that facility-based data on abortion-related SMO suffers a number of limitations.


Decrease in the abundance and viability of oceanic phytoplankton due to trace levels of complex mixtures of organic pollutants.
Long range atmospheric transport and deposition is a significant introduction pathway of organic pollutants to remote oceanic regions, leading to their subsequent accumulation in marine organisms. Persistent organic pollutants (POPs) bioconcentrate in planktonic food webs and these exert a biogeochemical control on the regional and global cycling of POPs. Therefore, an important issue is to determine whether the anthropogenic chemical perturbation of the biosphere introduced by the myriad of organic pollutants present in seawater influences phytoplankton abundance and productivity. The results reported here from five sets of experiments performed in the NE Atlantic Ocean show that there is a toxic effect induced by trace levels of complex mixtures of organic pollutants on phytoplankton oceanic communities. The levels of single pollutant, such as phenanthrene and pyrene, at which lethality of phytoplankton is observed are high in comparison to field levels. Complex mixtures of organic pollutants, however, have an important toxic effect on phytoplankton abundances, viability and concentrations of Chlorophyll a at pollutant concentrations 20-40 folds those found in the open ocean. The toxicity of these complex mixtures of organic pollutants exceeds by 10(3) times the toxicity expected for a single pollutant. Therefore, our results point out the need for a systematic investigation of the influence of complex mixtures of organic hydrophobic pollutants to oceanic phytoplankton communities, a perturbation not accounted for on previous assessments of anthropogenic pressures in the marine environment.


Multicopy FZF1 (SUL1) suppresses the sulfite sensitivity but not the glucose derepression or aberrant cell morphology of a grr1 mutant of Saccharomyces cerevisiae.
An ssu2 mutation in Saccharomyces cerevisiae, previously shown to cause sulfite sensitivity, was found to be allelic to GRR1, a gene previously implicated in glucose repression. The suppressor rgt1, which suppresses the growth defects of grr1 strains on glucose, did not fully suppress the sensitivity on glucose or nonglucose carbon sources, indicating that it is not strictly linked to a defect in glucose metabolism. Because the Cln1 protein was previously shown to be elevated in grr1 mutants, the effect of CLN1 overexpression on sulfite sensitivity was investigated. Overexpression in GRR1 cells resulted in sulfite sensitivity, suggesting a connection between CLN1 and sulfite metabolism. Multicopy FZF1, a putative transcription factor, was found to suppress the sulfite sensitive phenotype of grr1 strains, but not the glucose derepression or aberrant cell morphology. Multicopy FZF1 was also found to suppress the sensitivity of a number of other unrelated sulfite-sensitive mutants, but not that of ssu1 or met20, implying that FZF1 may act through Ssu1p and Met20p. Disruption of FZF1 resulted in sulfite sensitivity when the construct was introduced in single copy at the FZF1 locus in a GRR1 strain, providing evidence that FZF1 is involved in sulfite metabolism.


Inactivation of the Mycobacterium tuberculosis antigen 85 complex by covalent, allosteric inhibitors.
The rise of multidrug-resistant and totally drug-resistant tuberculosis and the association with an increasing number of HIV-positive patients developing tuberculosis emphasize the necessity to find new antitubercular targets and drugs. The antigen 85 (Ag85) complex from Mycobacterium tuberculosis plays important roles in the biosynthesis of major components of the mycobacterial cell envelope. For this reason, Ag85 has emerged as an attractive drug target. Recently, ebselen was identified as an effective inhibitor of the Ag85 complex through covalent modification of a cysteine residue proximal to the Ag85 active site and is therefore a covalent, allosteric inhibitor. To expand the understanding of this process, we have solved the x-ray crystal structures of Ag85C covalently modified with ebselen and other thiol-reactive compounds, p-chloromercuribenzoic acid and iodoacetamide, as well as the structure of a cysteine to glycine mutant. All four structures confirm that chemical modification or mutation at this particular cysteine residue leads to the disruption of the active site hydrogen-bonded network essential for Ag85 catalysis. We also describe x-ray crystal structures of Ag85C single mutants within the catalytic triad and show that a mutation of any one of these three residues promotes the same conformational change observed in the cysteine-modified forms. These results provide evidence for active site dynamics that may afford new strategies for the development of selective and potent Ag85 inhibitors.


Intraobserver and interobserver agreement in visual inspection for xanthochromia: implications for subarachnoid hemorrhage diagnosis, computed tomography validation studies, and the Walton rule.
Visual inspection for xanthochromia is used to diagnose subarachnoid hemorrhage (SAH), to validate computed tomography subarachnoid hemorrhage diagnosis and was used to determine the Walton rule. No study has assessed the reliability of xanthochromia. To determine intraobserver and interobserver xanthochromia agreement. Mock cerebrospinal fluid samples contained increasing concentrations of human oxyhemoglobin, bilirubin, and albumin. Non-color-blind observers randomly assessed samples against a white background twice under significantly differing illumination. Specimens were recorded as red, orange, yellow, or clear. Results were obtained for 26 observers (11 male, 15 female observers). We found that 19.2% of samples were misclassified: red, 11.7%; orange, 28.5%; yellow, 29.6%; and clear, 22.1% (χ = 213.2; P < .001). Of the yellow misclassifications, 88% were misclassified as clear. Female observers correctly classified samples significantly more frequently than male observers (P = .03). Intraobserver agreement differed significantly from expected for both male (χ = 105.6; P < .001) and female (χ = 99.9; P < .001) observers regardless of illumination. Interobserver agreement was poor regardless of sex (χ for male observers = 176.96, P < .001; χ for female observers = 182.69, P < .001) or illumination (χ for bright = 125.64, P < .001; χ for dark = 148.48, P < .001). Overall, there was 75% agreement in 46% of the tests and 90% agreement in only 36% of the tests. This simple laboratory study would be expected to maximize agreement relative to clinical practice. Although non-color-blind female observers significantly outperformed non-color-blind male observers, both intraobserver agreement and interobserver agreement for xanthochromia were prohibitively poor regardless of sex or illumination. Yellow was most frequently misclassified, 88% as clear (ie, true positives were commuted to false negatives). Xanthochromia is therefore highly unreliable for subarachnoid hemorrhage diagnosis and computed tomography validation. The Walton rule requires urgent clinical revalidation.


Cryptic contamination and phylogenetic nonsense.
Ancient human DNA has been treated cautiously ever since the problems related to this type of material were exposed in the early 1990s, but as sequential genetic data from ancient specimens have been key components in several evolutionary and ecological studies, interest in ancient human DNA is on the increase again. It is especially tempting to approach archaeological and anthropological questions through this type of material, but DNA from ancient human tissue is notoriously complicated to work with due to the risk of contamination with modern human DNA. Various ways of authenticating results based on ancient human DNA have been developed to circumvent the problems. One commonly used method is to predict what the contamination is expected to look like and then test whether the ancient human DNA fulfils this prediction. If it does, the results are rejected as contamination, while if it does not, they are often considered authentic. We show here that human contamination in ancient material may well deviate from local allele frequencies or the distributions to be found among the laboratory workers and archaeologists. We conclude that it is not reliable to authenticate ancient human DNA solely by showing that it is different from what would be expected from people who have handled the material.


SU-E-T-25: Real Time Simulator for Designing Electron Dual Scattering Foil Systems.
To create a user friendly, accurate, real time computer simulator to facilitate the design of dual foil scattering systems for electron beams on radiotherapy accelerators. The simulator should allow for a relatively quick, initial design that can be refined and verified with subsequent Monte Carlo (MC) calculations and measurements. The simulator consists of an analytical algorithm for calculating electron fluence and a graphical user interface (GUI) C++ program. The algorithm predicts electron fluence using Fermi-Eyges multiple Coulomb scattering theory with a refined Moliere formalism for scattering powers. The simulator also estimates central-axis x-ray dose contamination from the dual foil system. Once the geometry of the beamline is specified, the simulator allows the user to continuously vary primary scattering foil material and thickness, secondary scattering foil material and Gaussian shape (thickness and sigma), and beam energy. The beam profile and x-ray contamination are displayed in real time. The simulator was tuned by comparison of off-axis electron fluence profiles with those calculated using EGSnrc MC. Over the energy range 7-20 MeV and using present foils on the Elekta radiotherapy accelerator, the simulator profiles agreed to within 2% of MC profiles from within 20 cm of the central axis. The x-ray contamination predictions matched measured data to within 0.6%. The calculation time was approximately 100 ms using a single processor, which allows for real-time variation of foil parameters using sliding bars. A real time dual scattering foil system simulator has been developed. The tool has been useful in a project to redesign an electron dual scattering foil system for one of our radiotherapy accelerators. The simulator has also been useful as an instructional tool for our medical physics graduate students.


A frequency-agile terahertz-wave parametric oscillator.
We demonstrated a pulse-to-pulse frequency-tunable LiNbO3 terahertz-wave parametric oscillator, pumped with a Q-switched Nd:YAG laser. Rapid tuning from 1 to 2 THz, with random frequency accessibility, was achieved by rotating the pump beam angle using an optical beam scanner and a telescope.


Plastic debris and policy: Using current scientific understanding to invoke positive change.
Captain Charles Moore introduced the world to the "Great Pacific Garbage Patch" in the mid-1990s, and images of plastic debris in the oceans began to sweep the media. Since then, there has been increasing interest from scientists, the public, and policy makers regarding plastic debris in the environment. Today, there remains no doubt that plastic debris contaminates aquatic (marine and freshwater) habitats and animals globally. The growing scientific evidence demonstrates widespread contamination from plastic debris, and researchers are beginning to understand the sources, fate, and effects of the material. As new scientific understanding breeds new questions, scientists are working to fill data gaps regarding the fate and effects of plastic debris and the mechanisms that drive these processes. In parallel, policy makers are working to mitigate this contamination. The authors focus on what is known about plastic debris that is relevant to policy by reviewing some of the weight of evidence regarding contamination, fate, and effects of the material. Moreover, they highlight some examples of how science has already been used to inform policy change and mitigation and discuss opportunities for future linkages between science and policy to continue the relationship and contribute to effective solutions for plastic debris. Environ Toxicol Chem 2016;35:1617-1626. © 2016 SETAC.


PCO2 affects tracheal tone during apnea in anesthetized dogs.
We hypothesized that CO2, like hypoxia and withdrawal of pulmonary slowly adapting receptor input, would cause tracheal constriction during neural apnea (absence of phrenic activity). In seven anesthetized paralyzed dogs ventilated to neural apnea, we increased arterial PCO2 (PaCO2) in steps by adding CO2 to the inspirate while keeping ventilation constant. Increases in PaCO2 caused tracheal constriction during neural apnea in all dogs; 69 +/- 26 (SD)% of the change in tracheal diameter occurred during neural apnea. Average sensitivity of tracheal diameter to CO2 was 0.44 mm/Torr PaCO2. Our data suggest that central chemoreceptor inputs to brain stem neurons controlling smooth muscle of the extrathoracic airway bypass central mechanisms generating inspiration.


Thrombospondin-1 protects against pathogen-induced lung injury by limiting extracellular matrix proteolysis.
Acute lung injury is characterized by excessive extracellular matrix proteolysis and neutrophilic inflammation. A major risk factor for lung injury is bacterial pneumonia. However, host factors that protect against pathogen-induced and host-sustained proteolytic injury following infection are poorly understood. Pseudomonas aeruginosa (PA) is a major cause of nosocomial pneumonia and secretes proteases to amplify tissue injury. We show that thrombospondin-1 (TSP-1), a matricellular glycoprotein released during inflammation, dose-dependently inhibits PA metalloendoprotease LasB, a virulence factor. TSP-1-deficient (Thbs1-/-) mice show reduced survival, impaired host defense, and increased lung permeability with exaggerated neutrophil activation following acute intrapulmonary PA infection. Administration of TSP-1 from platelets corrects the impaired host defense and aberrant injury in Thbs1-/- mice. Although TSP-1 is cleaved into 2 fragments by PA, TSP-1 substantially inhibits Pseudomonas elastolytic activity. Administration of LasB inhibitor, genetic disabling of the PA type II secretion system, or functional deletion of LasB improves host defense and neutrophilic inflammation in mice. Moreover, TSP-1 provides an additional line of defense by directly subduing host-derived proteolysis, with dose-dependent inhibition of neutrophil elastase from airway neutrophils of mechanically ventilated critically ill patients. Thus, a host matricellular protein provides dual levels of protection against pathogen-initiated and host-sustained proteolytic injury following microbial trigger.


Breast cancer disparities and decision-making among U.S. women.
The impact of breast cancer is immense for all women, but the literature reveals an even greater impact on women of color and among socially and economically disadvantaged populations. Persistent differences in incidence and outcome are undoubtedly due to multiple factors, but one element in poor outcome may be treatment choice. Those treatments shown to be related to best outcomes are less likely to be chosen by certain groups of women. The effects of economic and cultural factors on breast cancer treatment choice have not been thoroughly explored; these factors must be understood if health care professionals are to intervene effectively to address disparities and improve breast cancer outcomes for all women. A review of the breast cancer literature was conducted in order to: (1) describe breast cancer disparities in the United States; (2) delineate factors that might contribute to those disparities; (3) assess possible mitigating factors for predominant causes; (4) begin to decide how health care interventions might allay the factors that contribute to disparities in breast cancer incidence and mortality. Breast cancer incidence and outcome disparities in the United States are due to multiple interacting factors. These include information about treatment, different types of treatment, the emotional context of decision-making, and patient preference for level of involvement. Treatment decision-making is complex. Health literacy and level of decision-making involvement, both embedded in social and economic reality, are key components in breast cancer treatment decision-making and may contribute to breast cancer disparities in the United States. Current models of shared decision-making may not be generalizable to all breast cancer patients. Optimal breast cancer outcomes for all women depend on culturally and ethnically appropriate professional support.


Ethnicity and Prostate Cancer in Southern Nigeria: A Preliminary Report.
The natural history of prostate cancer varies among patients. The aim of this study is to detect any variations in clinical and pathological characteristics of the tumor in patients from different ethnic groups in Southern Nigeria. Consecutive patients who presented with features of prostatic diseases at the Urology Units of University of Port Harcourt Teaching Hospital, Port Harcourt and Nnamdi Azikiwe University Teaching Hospital, Nnewi, were evaluated prospectively with history, physical examination, and relevant investigations using a proforma. Data obtained were collated and analyzed statistically using the Chi-square test and Microsoft Excel. Of 187 patients studied, 169 were analyzed. Eighty-six were Ibos, 31 Ijaws, 25 Ikwerres, and 12 Ogonis. Two were from each Etche, Urhobo, Opobo, and Effik; 4 from Andoni, and 3 Ibibio. Fifty-seven (66.3%) Ibos presented with the disease at higher ages (70-80 years) than 19 (61.3%) Ijaws and 11 (91.7%) Ogonis. These age differences were statistically significant with 95% and 99.9% confidence, respectively. All cases were adenocarcinomas. Clinical features, pattern of serum prostate-specific antigen levels, grades of the tumors, tumor metastases, and complications were similar for all ethnic groups. Although more Ibos had tumors with relatively more aggressive metastatic features, there was no statistical significance. Clinical and pathological features of adenocarcinoma of the prostate in Ibos, Ikwerres, Ijaws, and Ogonis were found to be similar. However, Ibos presented with the disease at older ages than Ijaws and Ogonis.


The antiviral alkaloid berberine ameliorates neuropathic pain in rats with peripheral nerve injury.
Neuropathic pain is a major public health problem. There is a need to develop safer and more effective analgesia compounds with less side effects. Berberine has been used to treat diarrhea and gastroenteritis due to its anti-microbial, anti-motility and anti-secretory properties. Berberine has also been reported to play an analgesic role in some pathological conditions of pain. However, the analgesic roles of berberine in neuropathic pain are still unclear. Therefore, this study aims to explore the analgesic effects of berberine in neuropathic pain. Partial sciatic nerve ligation (pSNL) was performed to create neuropathic pain model. Paw withdrawal responses to mechanical and thermal stimuli were measured using a set of electronic von Frey apparatus and hot plate, respectively. The time that rats spent licking, flinching and lifting its paw during 5 min following capsaicin application was recorded. mRNA and protein expression levels were examined by quantitative RT-PCR and western blot, respectively. Berberine administration (i.p.) increased both mechanical and thermal pain thresholds in a dose-dependent manner. Moreover, berberine administration reversed the mRNA and protein expression of TRPV1 in dorsal root ganglion neurons after peripheral nerve injury. In addition, berberine significantly inhibited capsaicin-induced pain behaviors. The amelioration of neuropathic pain by berberine may be associated with the down-regulation of TRPV1 in DRG of neuropathic pain rats. This study highlighted the potential of berberine in the treatment of neuropathic pain originated in the peripheral nervous system.


On-chip learning with analogue VLSI neural networks.
Results from simulations of weight perturbation as an on-chip learning scheme for analogue VLSI neural networks are presented. The limitations of analogue hardware are modelled as realistically as possible. Thus synaptic weight precision is defined according to the smallest change in the weight setting voltage which gives a measurable change at the output of the corresponding neuron. Tests are carried out on a hard classification problem constructed from mobile robot navigation data. The simulations show that the degradation in classification performance on a 500-pattern test set caused by the introduction of realistic hardware constraints is acceptable: with 8-bit weights, updated probabilistically and with a simplified output error criterion, the error rate increases by no more than 7% when compared with weight perturbation implemented with full 32-bit precision.


The eighth FIII domain of human fibronectin promotes integrin alpha5beta1 binding via stabilization of the ninth FIII domain.
Binding of the extracellular matrix molecule fibronectin to the integrin receptor alpha(5)beta(1) elicits downstream signaling pathways that modulate cell function. Fibronectin-alpha(5)beta(1) interaction occurs via the conserved RGD sequence in the tenth FIII (FIII10) domain of fibronectin. A synergistic site containing the sequence PHSRN in the adjacent FIII9 domain has also been identified. Here we investigate the function of the eighth FIII domain in integrin-mediated cell adhesion using a wide range of methods, including biochemical, biological, and biophysical assays of integrin binding, cell adhesion, and protein denaturation. Mutation of the FIII9 synergistic site (PHSRN to PHAAA) in FIII9-10 reduced the binding activity for integrin alpha(5)beta(1) to levels observed for FIII10 alone, but the corresponding mutant in FIII8-9-10 showed no loss of binding activity. Cell adhesion assays also demonstrated enhanced functional activity of constructs containing FIII8. Equilibrium chemical denaturation studies indicated that FIII8 confers conformational stability upon FIII9, but only if the exposed loops, PHSRN and VKNEED on FIII9 and FIII8, respectively, are intact. These results demonstrate that the loss of integrin binding activity, observed upon alteration of the PHSRN synergistic site of FIII9-10, results partly from a loss of conformational stability of FIII9. Our data suggest a mechanism for integrin alpha(5)beta(1)-fibronectin interaction, which in addition to the primary RGD binding event, involves a conformation-sensitive scanning by the integrin for accessible sites on the ligand, whereupon full activation of downstream signaling occurs.


Transcriptome-wide single nucleotide polymorphisms (SNPs) for abalone (Haliotis midae): validation and application using GoldenGate medium-throughput genotyping assays.
Haliotis midae is one of the most valuable commercial abalone species in the world, but is highly vulnerable, due to exploitation, habitat destruction and predation. In order to preserve wild and cultured stocks, genetic management and improvement of the species has become crucial. Fundamental to this is the availability and employment of molecular markers, such as microsatellites and single nucleotide (SNPs). Transcriptome sequences generated through sequencing-by-synthesis technology were utilized for the in vitro and in silico identification of 505 putative SNPs from a total of 316 selected contigs. A subset of 234 SNPs were further validated and characterized in wild and cultured abalone using two Illumina GoldenGate genotyping assays. Combined with VeraCode technology, this genotyping platform yielded a 65%-69% conversion rate (percentage polymorphic markers) with a global genotyping success rate of 76%-85% and provided a viable means for validating SNP markers in a non-model species. The utility of 31 of the validated SNPs in population structure analysis was confirmed, while a large number of SNPs (174) were shown to be informative and are, thus, good candidates for linkage map construction. The non-synonymous SNPs (50) located in coding regions of genes that showed similarities with known proteins will also be useful for genetic applications, such as the marker-assisted selection of genes of relevance to abalone aquaculture.


Recent approaches in food bio-preservation - a review.
Bio-preservation is a technique of extending the shelf life of food by using natural or controlled microbiota or antimicrobials. The fermentation products as well as beneficial bacteria are generally selected in this process to control spoilage and render pathogen inactive. The special interest organism or central organism used for this purpose is lactic acid bacteria (LAB) and their metabolites. They are capable to exhibit antimicrobial properties and helpful in imparting unique flavour and texture to the food products. The major compounds produced by LAB are bacteriocin, organic acids and hydrogen peroxide. Bacteriocin is peptides or proteins with antimicrobial activity. On the basis of size, structure and post-translational modification, bacteriocin is divided into four different classes. Due to non-toxic, non-immunogenic, thermo-resistance characteristics and broad bactericidal activity, LAB bacteriocins are considered good bio-preservative agents. The most common LAB bactriocin is nisin which has wider applications in food industry and has been Food and Drug Administration (FDA) approved. Nisin and other bacteriocin are being used in vegetables products, dairy and meat industries. Apart from LAB metabolites, bacteriophages and endolysins has promising role in food processing, preservation and safety. Bacteriocins and endolysins are more suitable for DNA shuffling and protein engineering to generate highly potent variants with expanded activity spectrum. Genetically modified bacteriophages may also be helpful in bio-preservation, however; their safety issues must be addressed properly before selection as bio-preservative agent.


PEDOT:PSS interfaces stabilised using a PEGylated crosslinker yield improved conductivity and biocompatibility.
The rapidly expanding fields of bioelectronics, and biological interfaces with electronic sensors and stimulators, are placing an increasing demand on candidate materials to serve as robust surfaces that are both biocompatible, stable and electroconductive. Amongst conductive polymers, poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) is a promising material in biomedical research due to its appropriate stability and high conductivity, however its intrinsic solubility requires a crosslinking process that can limit its conductivity and biocompatibility. Poly(ethylene glycol) is known to be a suitably anti-immunogenic moiety and its derivatives have been widely used for biomedical applications. In this study we investigate the application of poly(ethylene glycol)diglycidyl ether (PEGDE) as an effective crosslinker and conductive filler for PEDOT:PSS. From our interpretation of XPS analysis we hypothesise that the crosslinking reaction is occurring via the epoxy ring of PEGDE interacting with the sulfonic groups of excel PSS chains, which reaches a saturation at 3 w/v% PEGDE concentration. PEGDE crosslinked films did not disperse in aqueous environments, had enhanced electrical conductivity and imparted a significant degree of hydrophilicity to PEDOT:PSS films. This hydrophilicity and the presence of biocompatible PEGDE led to good cell viability and a significantly increased degree of cell spreading on PEDOT:PSS films. In comparison to widely reported chemical crosslinking via glycidoxy propyltrimethoxysilane (GOPS), this original crosslinking yields a highly hydrophilic 2D film substrate with increased electroconductive and biocompatibility properties, resulting in a next-generation formulation for bioengineering applications.


Biological similarity of enterotoxins of Vibrio cholerae serotypes other than type 1 to cholera toxin and Escherichia coli heat-labile enterotoxin.
Vibrio cholerae serotypes other than type 1, the so-called NAG vibrios, have been recognised as an important cause of diarrhoea. A few of them have been shown to produce an enterotoxin similar, immunologically and physiologically, to cholera toxin; and cholera toxin has been shown to be structurally, functionally and immunologically identical with Escherichia coli heat-labile toxin (LT). The present investigation has demonstrated biological similarities among cholera toxin, E. coli LT and enterotoxins produced by strains of V. cholerae of 59 serotypes other than 1, in the biological models, rabbit ileal loops and rabbit skin. Culture filtrates of almost all the strains were neutralised completely and all filtrates showed some neutralisation, in enterotoxic action and increase of permeability, by cholera antitoxin and E. coli LT antiserum. The partial neutralisation observed in a few strains was probably due to high concentrations of identical toxin rather than the presence of other toxic substances.


Subacute cardiac toxicity following autologous haematopoietic stem cell transplantation in patients with normal cardiac function.
To investigate subacute cardiac toxicity in patients with normal baseline cardiac function following autologous haematopoietic stem cell transplantation. Prospective observational study. Thirty-two consecutive patients (mean (SD) age 60 (11) years) with normal left ventricular ejection fraction (LVEF >or=50%) undergoing autologous haematopoietic stem cell transplantation were studied. Transthoracic echocardiography (including colour tissue Doppler imaging-derived myocardial velocities, strain and strain rates), troponin-T and B-type natriuretic peptide (BNP) and clinical details were recorded at baseline, after conditioning chemotherapy and serially over 6 weeks from the day of transplantation. The mean (SD) LVEF at baseline was 62 (6)% and decreased to 55 (16)%, 6 weeks after transplantation (p = 0.007). Cardiac toxicity (>or=10% absolute decline of LVEF to an LVEF <or=50%) developed in 10 (31%) patients within 17 (8) days of transplantation and was usually reversible. In these 10 patients, the nadir LVEF was 39 (12)%. Two patients developed severe clinical pulmonary oedema, one episode of which was fatal. Troponin-T was mildly raised in only three of the patients with cardiac toxicity. Peak BNP values were similar in patients with or without post-transplant cardiac toxicity (149 (100) vs 196 (178) pg/ml, p = 0.43). Multivariate Cox proportional hazard regression identified baseline mitral annular systolic velocity as the only independent predictor of cardiac toxicity (hazard ratio 0.42, 95% confidence interval 0.20 to 0.86, p = 0.02). Subacute cardiac toxicity is common after autologous haematopoietic stem cell transplantation, even in patients with apparently normal left ventricular function. Anticipation of the period of greatest risk and recognition of patients with subclinical myocardial dysfunction may prevent clinical heart failure.


Use of Growing Cells of Pseudomonas aeruginosa for Synthesis of the Natural Vanillin via Conversion of Isoeugenol.
The great demand of people for consumption of natural additives resulted in producing natural vanillin. There are plant sources and chemical procedures for vanillin production but microbial bioconversions are being sought as a suitable alternative. In the present work, the ability to produce vanillin from isoeugenol was screened using growing cultures of various bacteria. Among the 56 strains of bacteria isolated from the soil environments of Iran, a Gram-negative rod designated as strain ISPC2 showed the capability of promoting the formation of high amounts of vanillin when grown in the presence of isoeugenol. On the basis of morphological and physiochemical characteristics and 16S ribosomal ribonucleic acid (rRNA) gene sequence analysis, the isolate was identified as Pseudomonas aeruginosa ISPC2. Vanillin formation was analyzed by GC/FID. In the presence of isoeugenol, a growing culture of P. aeruginosa ISPC2 produced 1.62 g/L vanillin (molar yield of 17.3%) after a 72 h reaction at 30°C and 200 rpm. This proposed procedure is an alternative approach to obtain vanillin in an environmentally friendly way. Further studies for standardization and optimization for higher yield of vanillin production, needs to be investigated.


A biosynthetic strategy for re-engineering the Staphylococcus aureus cell wall with non-native small molecules.
Staphylococcus aureus (S. aureus) is a Gram-positive bacterial pathogen that has emerged as a major public health threat. Here we report that the cell wall of S. aureus can be covalently re-engineered to contain non-native small molecules. This process makes use of endogenous levels of the bacterial enzyme sortase A (SrtA), which ordinarily functions to incorporate proteins into the bacterial cell wall. Thus, incubation of wild-type bacteria with rationally designed SrtA substrates results in covalent incorporation of functional molecular handles (fluorescein, biotin, and azide) into cell wall peptidoglycan. These conclusions are supported by data obtained through a variety of experimental techniques (epifluorescence and electron microscopy, biochemical extraction, and mass spectrometry), and cell-wall-incorporated azide was exploited as a chemical handle to perform an azide-alkyne cycloaddition reaction on the bacterial cell surface. This report represents the first example of cell wall engineering of S. aureus or any other pathogenic Gram-positive bacteria and has the potential for widespread utility.


Visiting Nurses' Perspectives on Practices to Achieve End-of-Life Cancer Patients' Wishes for Death at Home: A Qualitative Study.
This study aimed to clarify visiting nurses' perspectives on critical practices to ensure they could advocate for patients who prefer to die at home. Sixteen nurses, working at home-visit nursing agencies in Japan, participated in this study. Data were generated by interviews with the nurses and participant observations from nursing home-visits for six end-of-life cancer patients and were analyzed using content analysis. Five themes emerged: (1) nursing assessment, (2) support for comfortable daily life of the patient and their family, (3) advocating for the patient's views about continuing homecare until death, (4) supporting the patient's preparedness for death, and (5) coordination with other health professionals and related facilities for a comfortable environment for the patient. In addition, the nurses sometimes used humorous responses to death-related work to change the patient's melancholy thoughts. The present study found that the participants advocated for the patient's views about continuing homecare until death while coordinating views between the patient and their family; they further supported the patient's daily life while helping them prepare for death to achieve their wish for death at home. In addition, our study uncovered the visiting nurses' unconscious practical wisdom of using humorous responses to death-related work to alleviate the patients' feelings of hopelessness. To develop practical wisdom for using humor effectively in end-of-life care, nurses need to verbalize unconscious practices, and accumulate empirical knowledge about nursing interventions using humor, including cultural attitudes, through case study analysis.


Study of a classification algorithm for AIEC identification in geographically distinct E. coli strains.
Adherent-invasive Escherichia coli (AIEC) have been extensively implicated in Crohn's disease pathogenesis. Currently, AIEC is identified phenotypically, since no molecular marker specific for AIEC exists. An algorithm based on single nucleotide polymorphisms was previously presented as a potential molecular tool to classify AIEC/non-AIEC, with 84% accuracy on a collection of 50 strains isolated in Girona (Spain). Herein, our aim was to determine the accuracy of the tool using AIEC/non-AIEC isolates from different geographical origins and extraintestinal pathogenic E. coli (ExPEC) strains. The accuracy of the tool was significantly reduced (61%) when external AIEC/non-AIEC strains from France, Chile, Mallorca (Spain) and Australia (82 AIEC, 57 non-AIEC and 45 ExPEC strains in total) were included. However, the inclusion of only the ExPEC strains showed that the tool was fairly accurate at differentiating these two close pathotypes (84.6% sensitivity; 79% accuracy). Moreover, the accuracy was still high (81%) for those AIEC/non-AIEC strains isolated from Girona and Mallorca (N = 63); two collections obtained from independent studies but geographically close. Our findings indicate that the presented tool is not universal since it would be only applicable for strains from similar geographic origin and demonstrates the need to include strains from different origins to validate such tools.


Relation between physical activity and blood pressure levels in young Greek adolescents: the Leontio Lyceum Study.
Elevated blood pressure (BP) during childhood and adolescence is not so rare and increases the risk of hypertension in adulthood. Physical activity (PA) is considered a key component for the prevention and treatment of hypertension in children and adolescents. Thus, the purpose of our study was to assess the level of PA in Greek adolescents and its relation to BP levels. The study included 496 students aged 12-17 years who were enrolled on a volunteer basis. All children were subjected to BP measurement on two different occasions during a routine school day. Demographic, socio-economic and lifestyle characteristics were assessed by means of a standard questionnaire. Information on the frequency and duration of PA and on the amount of time spent on sedentary activities was obtained by the short-form self-administered questionnaire International Physical Activity Questionnaire-IPAQ. The study population was divided in three groups according to the score achieved in IPAQ questionnaire: low PA (n = 39), moderate PA (n = 230) and high PA (n = 231). Children with high PA compared to those with low PA exhibited higher systolic BP and pulse pressure levels, greater prevalence of prehypertension/hypertension status, while heart rate was significantly lower as the level of PA rose (P < 0.05 for all cases). Intense PA was positively correlated to systolic BP (r = 0.139, P = 0.003) and pulse pressure (r = 0.22, P = 0.0001). Intense PA is associated with higher systolic BP, pulse pressure levels and lower heart rate in healthy young adolescents. PA should be practiced at a moderate intensity level in everyday life.


Clinical severity of psoriasis in last 20 years of PUVA study.
To assess the severity of psoriasis over time. We analyzed the results of structured dermatologic examinations administered over a 20-year period beginning 10 years after study enrollment. The PUVA [psoralen-UV-A] Follow-up Study, which is a prospective cohort study. The analyses were restricted to 815 patients (83.2% of those eligible) who underwent at least 2 of 4 possible examinations between 1985 and 2005. A 4-point physician global assessment (PGA). The distribution of the PGA levels in the study group did not change significantly over time, except that in 2005 more patients had no psoriasis compared with patients who underwent examinations in the previous study years (9.6% vs < 5.1%, P < .03). The PGA level changed more than 1 level between examinations in only 14% of patients. Multistate Markov models estimated that patients had a likelihood of about 80% to remain at the same PGA level 1 year later. After 10 years, this likelihood varied between 19% and 53%, depending on the PGA level. Except for patients who were clear of disease at baseline, on average patients had about 1 year without psoriasis over 20 years. On average, individuals with moderate to severe disease remained at these levels for 11 or more years. Conclusion Three decades after a large and diverse group of patients sought a cure for their psoriasis, consistent control of their psoriasis often had not been achieved.


Identification of the 2-hydroxyglutarate and isovaleryl-CoA dehydrogenases as alternative electron donors linking lysine catabolism to the electron transport chain of Arabidopsis mitochondria.
The process of dark-induced senescence in plants is relatively poorly understood, but a functional electron-transfer flavoprotein/electron-transfer flavoprotein:ubiquinone oxidoreductase (ETF/ETFQO) complex, which supports respiration during carbon starvation, has recently been identified. Here, we studied the responses of Arabidopsis thaliana mutants deficient in the expression of isovaleryl-CoA dehydrogenase and 2-hydroxyglutarate dehydrogenase to extended darkness and other environmental stresses. Evaluations of the mutant phenotypes following carbon starvation induced by extended darkness identify similarities to those exhibited by mutants of the ETF/ETFQO complex. Metabolic profiling and isotope tracer experimentation revealed that isovaleryl-CoA dehydrogenase is involved in degradation of the branched-chain amino acids, phytol, and Lys, while 2-hydroxyglutarate dehydrogenase is involved exclusively in Lys degradation. These results suggest that isovaleryl-CoA dehydrogenase is the more critical for alternative respiration and that a series of enzymes, including 2-hydroxyglutarate dehydrogenase, plays a role in Lys degradation. Both physiological and metabolic phenotypes of the isovaleryl-CoA dehydrogenase and 2-hydroxyglutarate dehydrogenase mutants were not as severe as those observed for mutants of the ETF/ETFQO complex, indicating some functional redundancy of the enzymes within the process. Our results aid in the elucidation of the pathway of plant Lys catabolism and demonstrate that both isovaleryl-CoA dehydrogenase and 2-hydroxyglutarate dehydrogenase act as electron donors to the ubiquinol pool via an ETF/ETFQO-mediated route.


New and notable ion-channels in the sarcoplasmic/endoplasmic reticulum: do they support the process of intracellular Ca²⁺ release?
Intracellular Ca(2+) release through ryanodine receptor (RyR) and inositol trisphosphate receptor (IP3 R) channels is supported by a complex network of additional proteins that are located in or near the Ca(2+) release sites. In this review, we focus, not on RyR/IP3 R, but on other ion-channels that are known to be present in the sarcoplasmic/endoplasmic reticulum (ER/SR) membranes. We review their putative physiological roles and the evidence suggesting that they may support the process of intracellular Ca(2+) release, either indirectly by manipulating ionic fluxes across the ER/SR membrane or by directly interacting with a Ca(2+) -release channel. These channels rarely receive scientific attention because of the general lack of information regarding their biochemical and/or electrophysiological characteristics makes it difficult to predict their physiological roles and their impact on SR Ca(2+) fluxes. We discuss the possible role of SR K(+) channels and, in parallel, detail the known biochemical and biophysical properties of the trimeric intracellular cation (TRIC) proteins and their possible biological and pathophysiological roles in ER/SR Ca(2+) release. We summarise what is known regarding Cl(-) channels in the ER/SR and the non-selective cation channels or putative 'Ca(2+) leak channels', including mitsugumin23 (MG23), pannexins, presenilins and the transient receptor potential (TRP) channels that are distributed across ER/SR membranes but which have not yet been fully characterised functionally.


Carnosol protects against spinal cord injury through Nrf-2 upregulation.
Carnosol is an ortho-diphenolic diterpene with excellent antioxidant potential. The present study was designed to identify the protective role of carnosol against spinal cord injury (SCI)-induced oxidative stress and inflammation in Wistar rats. In the present study, oxidative stress status was determined through estimating total antioxidant capacity, total oxidant status, lipid peroxide content, protein carbonyl and sulfhydryl levels, reactive oxygen species (ROS), antioxidant status (superoxide-dismutase, catalase, glutathione, glutathione peroxidase, glutathione-S-transferase). Inflammatory effects were determined by analyzing the expression of NF-κB and COX-2 through Western blot analysis. Further, carnosol-mediated redox homeostasis was analyzed by determining p-AKT and Nrf-2 levels. SCI resulted in a significant increase in oxidative stress status through increased ROS generation, total oxidant levels, lipid peroxide content, protein carbonyl and sulfhydryl levels. The antioxidant status in SCI rats was significantly reduced, indicating imbalance in redox status. In addition, the expression of NF-κB and COX-2 was significantly upregulated, while p-AKT and Nrf-2 levels were downregulated in SCI rats. However, treatment with carnosol showed a significant enhancement in the antioxidant status with concomitant decline in oxidative stress parameters. Further, carnosol treatment regulated the key proteins in inflammation and redox status through significant downregulation of NF-κB and COX-2 levels and upregulation of p-AKT and Nrf-2 expression. Thus, the present study shows for the first time on the protective role of carnosol against SCI-induced oxidative stress and inflammation through modulating NF-κB, COX-2 and Nrf-2 levels in Wistar rats.


Effects of Xiaoqinglong decoction on gene expression profiles in a rat chronic obstructive pulmonary disease model.
Xiaoqinglong decoction (XQLD) has been used for centuries in Asia to effectively treat patients with chronic obstructive pulmonary disease (COPD). However, its mechanisms remain unknown. To elucidate this problem, we analyzed the effects of XQLD on gene expressions profiles in COPD rats. In the study, 20 male Wistar rats were injected with lipopolysaccharide (LPS), exposed to cigarette smoke and kept at -20°C for 5 min/day for a successive 8 days to establish COPD animals. Trachea ultramicrostructure and histomorphology were observed to determine whether these models were established successfully. Gene expression profiles were detected using cDNA microarrays. We found 56 differentially expressed genes associated with COPD progression, including 32 up-regulated genes and 24 down-regulated genes. These genes were confirmed to be involved in immune and inflammation reactions, metabolism, cell transportation and the cell cycle, signal transduction and gene regulation. Comparison of gene expression between the therapy group and control group showed that there were only 11 differentially expressed genes, including 5 up-regulated genes and 6 down-regulated genes. We concluded that XQLD had therapeutic effects in COPD rats by affecting gene expression. Pharmacological or targeted expression of some genes might be found useful as novel therapy in COPD management.


Downstream effects on human low density lipoprotein of homocysteine exported from endothelial cells in an in vitro system.
A model system is presented using human umbilical vein endothelial cells (HUVECs) to investigate the role of homocysteine (Hcy) in atherosclerosis. HUVECs are shown to export Hcy at a rate determined by the flux through the methionine/Hcy pathway. Additional methionine increases intracellular methionine, decreases intracellular folate, and increases Hcy export, whereas additional folate inhibits export. An inverse relationship exists between intracellular folate and Hcy export. Hcy export may be regulated by intracellular S-adenosyl methionine rather than by Hcy. Human LDLs exposed to HUVECs exporting Hcy undergo time-related lipid oxidation, a process inhibited by the thiol trap dithionitrobenzoate. This is likely to be related to the generation of hydroxyl radicals, which we show are associated with Hcy export. Although Hcy is the major oxidant, cysteine also contributes, as shown by the effect of glutamate. Finally, the LDL oxidized in this system showed a time-dependent increase in uptake by human macrophages, implying an upregulation of the scavenger receptor. These results suggest that continuous export of Hcy from endothelial cells contributes to the generation of extracellular hydroxyl radicals, with associated oxidative modification of LDL and incorporation into macrophages, a key step in atherosclerosis. Factors that regulate intracellular Hcy metabolism modulate these effects.


Rotarod training in mice is associated with changes in brain structure observable with multimodal MRI.
The brain has been shown to remain structurally plastic even throughout adulthood. However, little is known how motor-skill training affects different MRI modalities in the adult mouse brain. The aim of this study is to investigate whether rotarod training, a simple motor training task taken from the standard test battery, is associated with structural plasticity observable with different MRI modalities in adult C57BL/6 mice. The rotarod is a standard test that taxes motor coordination and balance. We use T2-weighted MRI followed by deformation-based morphometry to assess local volume and fractional anisotropy (FA) derived from diffusion MRI to assess microstructure ex-vivo. Using deformation-based morphometry we found that the hippocampus, frontal cortex and amygdala are larger in rotarod-trained mice compared to untrained controls. Surprisingly, the cerebellum and white matter in the corpus callosum underlying the primary motor cortex are smaller after training. We also found that the volume of the motor cortex is positively correlated with better rotarod performance. Diffusion imaging indicates group differences and behavioral correlations with FA, a measure of microstructure. Trained mice have higher FA in the hippocampus. Better rotarod performance is associated with higher FA in the hippocampus and lower FA in the primary visual cortex. This is the first study to reveal the substantial structural reorganization of the adult mouse brain following only a relatively brief period of motor-skill training by using complementary measures of microstructure and volume.


The scope and severity of white-nose syndrome on hibernating bats in North America.
Assessing the scope and severity of threats is necessary for evaluating impacts on populations to inform conservation planning. Quantitative threat assessment often requires monitoring programs that provide reliable data over relevant spatial and temporal scales, yet such programs can be difficult to justify until there is an apparent stressor. Leveraging efforts of wildlife management agencies to record winter counts of hibernating bats, we collated data for 5 species from over 200 sites across 27 U.S. states and 2 Canadian provinces from 1995 to 2018 to determine the impact of white-nose syndrome (WNS), a deadly disease of hibernating bats. We estimated declines of winter counts of bat colonies at sites where the invasive fungus that causes WNS (Pseudogymnoascus destructans) had been detected to assess the threat impact of WNS. Three species undergoing species status assessment by the U.S. Fish and Wildlife Service (Myotis septentrionalis, Myotis lucifugus, and Perimyotis subflavus) declined by more than 90%, which warrants classifying the severity of the WNS threat as extreme based on criteria used by NatureServe. The scope of the WNS threat as defined by NatureServe criteria was large (36% of Myotis lucifugus range) to pervasive (79% of Myotis septentrionalis range) for these species. Declines for 2 other species (Myotis sodalis and Eptesicus fuscus) were less severe but still qualified as moderate to serious based on NatureServe criteria. Data-sharing across jurisdictions provided a comprehensive evaluation of scope and severity of the threat of WNS and indicated regional differences that can inform response efforts at international, national, and state or provincial jurisdictions. We assessed the threat impact of an emerging infectious disease by uniting monitoring efforts across jurisdictional boundaries and demonstrated the importance of coordinated monitoring programs, such as the North American Bat Monitoring Program (NABat), for data-driven conservation assessments and planning.


Fabricating centimeter-scale high quality factor two-dimensional periodic photonic crystal slabs.
We present a fabrication route for centimeter-scale two-dimensional defect-free photonic crystal slabs with quality factors bigger than 10,000 in the visible, together with a unique way to quantify their quality factors. We fabricate Si(3)N(4) photonic crystal slabs, and perform an angle-resolved reflection measurement. This measurement data is used to retrieve the quality factors of the slabs by fitting it to a model based on temporal coupled-mode theory. The macroscopic nature of the structure and the high quality factors of their resonances could open up new opportunities for realizing efficient macroscale optoelectronic devices such as sensors, lasers, and energy harvesting systems.


Minimally invasive ultrasound-guided technique for central venous catheterization via the external jugular vein in pigs.
To describe an ultrasound-guided technique for central venous catheter placement via the external jugular vein (EJV) in pigs. 96 healthy Landrace-Poland China barrows (approx 16 weeks old with a mean weight of 70 kg). Pigs were anesthetized. With ultrasound guidance, a needle was inserted into the EJV without a large incision or cutdown procedure. A guidewire was inserted through the needle into the vein. A modified Seldinger technique was used to advance a catheter into the vessel until the tip was in the cranial vena cava near the right atrium. A trocar was used to create a tunnel through the subcutaneous tissues from the catheter insertion site to between the dorsal borders of the scapulae. The free end of the catheter was passed through that tunnel. An extension was attached to the catheter and secured to the skin. Pigs were euthanized and underwent necropsy at completion of the study for which they were catheterized. Central venous catheters were successfully placed in all 96 pigs and facilitated collection of serial blood samples with minimal stress. Catheters remained in place for a mean of 6 days (range, 4 to 10 days). Necropsy revealed abscesses along the subcutaneous catheter tract in 9 pigs. Twenty pigs had histologic evidence of phlebitis and fibroplasia in the cranial vena cava. The described technique, in combination with extensive socialization, allowed serial collection of blood samples with minimal stress and restraint and is an alternative to surgical cutdown procedures for catheter placement.


Programming effects of maternal stress on the circadian system of adult offspring.
Maternal stress has long-lasting influences on the brain functions of offspring, and several brain regions have been proposed to mediate such programming. Although perinatal programming of crosstalk between the circadian and stress systems has been proposed, the functional consequences of prenatal stress on the circadian system and the underlying mechanisms remain largely unknown. Therefore, we investigated whether exposing pregnant mice to chronic restraint stress had prolonged effects on the suprachiasmatic nucleus (SCN), which bears the central pacemaker for mammalian circadian rhythms, of offspring. SCN explants from maternally stressed mice exhibited altered cyclic expression patterns of a luciferase reporter under control of the mouse Per1 promoter (mPer1::LUC), which manifested as a decreased amplitude and impaired stability of the rhythm. Bioluminescence imaging at the single-cell level subsequently revealed that impaired synchrony among individual cells was responsible for the impaired rhythmicity. These intrinsic defects appeared to persist during adulthood. Adult male offspring from stressed mothers showed advanced-phase behavioral rhythms with impaired stability as well as altered clock gene expression in the SCN. In addition to affecting the central rhythm, maternal stress also had prolonged influences on the circadian characteristics of the adrenal gland and liver, as determined by circulating corticosterone levels and hepatic glycogen content, and on canonical clock gene mRNA expression in those tissues. Taken together, our findings suggest that the SCN is a key target of the programming effects of maternal stress. The widespread effects of circadian disruptions caused by a misprogrammed clock may have further impacts on metabolic and mental health in later life.


Biotransformation of green tea polyphenols and the biological activities of those metabolites.
Green tea ( Camellia sinensis, Theaceae) and its major polyphenol constituents, the catechins, have been reported to have many health benefits including the prevention of cancer and heart disease. Many mechanisms of action have been proposed based on in vitro models; however, the importance of most of these mechanisms remains to be determined in vivo. The bioavailability and biotransformation of tea catechins play a key role in determining the importance of various mechanisms in vivo. Likewise, the biological activity and bioavailability of tea catechin metabolites, an understudied area, are important in understanding the potential beneficial effects of tea. In this article, we review the data available on the biotransformation of the tea catechins and the limited data set available on the biological activities of the catechin metabolites. Careful interpretation of available data, carefully designed animal experiments, and integration of bioavailability and biological activity data are needed if the disease preventive activity of tea is to be understood. We hope this article will spark research efforts on some of the important questions regarding tea polyphenol bioavailability, biotransformation, and the biological activities of tea catechin metabolites.


The Vulnerability Scan, a Web Tool to Increase Institutional Biosecurity Resilience.
The importance of vigilance within organizations working with high-risk biological material receives increasing attention. However, an in-depth and comprehensive tool, dedicated to increase awareness of potential risks and to assess an organization's current biosecurity vulnerabilities, has not been available yet. We developed the "Biosecurity Vulnerability Scan," a web tool that identifies biosecurity gaps in an organization based on eight biosecurity pillars of good practice. Although the tool aims primarily to assist biosafety and biosecurity officers, it can also be useful to researchers working with dangerous pathogens, their principal investigators, management, or those responsible for security issues in the life sciences. Results are only stored locally and are provided in an "overview report," which includes information on relevant risks and control measures. This can support well-substantiated decision-making on strengthening biosecurity measures within a specific organization. With this article, we aim to support institutes to increase their overall security resilience and to improve institutional biosecurity in particular by providing practical recommendations. The Biosecurity Vulnerability Scan is available at www.biosecurityvulnerabilityscan.nl.


Immunoisolation effect of polyvinyl alcohol (PVA) macroencapsulated islets in type 1 diabetes therapy.
Islet transplantation has shown great success in the treatment of type 1 diabetes since the Edmonton protocol was established. However, it still has two major problems to overcome: the lack of organ donors and the side effects of immunosuppression. Encapsulated islets have emerged as a potential option for islet transplantation because it can, at least partly, overcome these two problems. Wistar rat islets suspended in 3% polyvinyl alcohol (PVA) hydrogel were frozen-thawed to make macroencapsulated islets (MEIs). The recovery rate, insulin content, and morphological change in culture medium with/without fresh human plasma (FHP) were measured in MEIs and free islets in vitro. In vivo, MEIs of either Wistar or Lewis rats were transplanted into the peritoneal cavity of streptozotocin (STZ)-induced diabetic Lewis rats and nonfasting blood glucose (NFBG), body weight, and histological evaluations were processed. FHP destroyed rat free islets but did not affect the islet morphology, islet recovery rate, or insulin content of rat MEIs. The transplantation of MEIs decreased the NFBG level and prevented body weight loss without a significant difference between the donor strains. Insulin-positive islets were observed in PVA MEIs 24 weeks after allotransplantation. These results suggest that PVA MEIs may be used as a cure for type 1 diabetes.


Beyond Depression: Estimating 12-Months Prevalence of Passive Suicidal Ideation in Mid- and Late-Life in the Health and Retirement Study.
To provide valid estimates of the 12-month prevalence of passive suicidal ideation among older adults, without conditioning on depression status, using the Health and Retirement Study (HRS). Data come from the 2012 HRS (n = 17,434) and 2004/5 Baltimore Epidemiologic Catchment Area (ECA) Study (n = 755). In the HRS, passive suicidal ideation (i.e., thought a lot about death-your own, someone else's, or death in general) is only assessed on respondents who reported dysphoria/anhedonia; in the ECA, ideation is assessed on all respondents, regardless of depression. We compare two approaches to estimating the 12-month prevalence of passive suicidal ideation in the HRS without conditioning on depression symptoms: 1) a probit selection model within the HRS, and 2) a prediction model developed using appended ECA data applied to the HRS. Using observed data alone on those who screened positive for depression, 6% of older adults reported passive suicidal ideation in the past year. Depending on the approach used, between 5.4% and 9.2% of HRS respondents who screened negative for depression would have reported passive suicidal ideation had they been assessed. Correcting for this selection bias, between 10.9% and 13.4% of U.S. adults over age 50 experienced passive suicidal ideation in 2012. Population surveillance of suicidal ideation among older adults is biased by survey approaches that only assess ideation in the context of depression.


On-site analysis of arsenic in groundwater using a microfabricated gold ultramicroelectrode array
Rapid on-site analysis of arsenic in groundwater was achieved with a small battery-powered unit in conjunction with a microfabricated gold ultramicroelectrode array (Au-UMEA). The sensor, consisting of 564 UME disks with a unique gold surface created by electron beam evaporation, was demonstrated to be highly sensitive to low-ppb As3+ using square wave anodic stripping voltammetry. The influence of the square wave frequency, pulse amplitude, and deposition potential on the arsenic peak stripping current was investigated. Varying those theoretical parameters yielded results surprisingly similar to those for the thin Hg film case. The performance of the Au-UMEA was evaluated for reproducibility and reliability. Three stability tests showed an average relative standard deviation of 2.5% for 15 consecutive runs. Limits of detection were investigated, and 0.05 ppb As3+ could be measured while maintaining a S/N of 3:1. Interference studies were performed in the presence of 50-500 ppb of Cu2+, Hg2+, and Pb2+. On-site analysis of groundwater containing arsenic was performed with a small battery-powered potentiostat. Quantification was done through standard additions, and these results were compared to the standard EPA methodology.


Measuring social-ecological dynamics behind the generation of ecosystem services.
The generation of ecosystem services depends on both social and ecological features. Here we focus on management, its ecological consequences, and social drivers. Our approach combined (1) quantitative surveys of local species diversity and abundance of three functional groups of ecosystem service providers (pollinators, seed dispersers, and insectivores) with (2) qualitative studies of local management practices connected to these services and their underlying social mechanisms, i.e., institutions, local ecological knowledge, and a sense of place. It focused on the ecology of three types of green areas (allotment gardens, cemeteries, and city parks) in the city of Stockholm, Sweden. These are superficially similar but differ considerably in their management. Effects of the different practices could be seen in the three functional groups, primarily as a higher abundance of pollinators in the informally managed allotment gardens and as differences in the composition of seed dispersers and insectivores. Thus, informal management, which is normally disregarded by planning authorities, is important for ecosystem services in the urban landscape. Furthermore, we suggest that informal management has an important secondary function: It may be crucial during periods of instability and change as it is argued to promote qualities with potential for adaptation. Allotment gardeners seem to be the most motivated managers, something that is reflected in their deeper knowledge and can be explained by a sense of place and management institutions. We propose that co-management would be one possible way to infuse the same positive qualities into all management and that improved information exchange between managers would be one further step toward ecologically functional urban landscapes.


Autism-Associated Insertion Mutation (InsG) of Shank3 Exon 21 Causes Impaired Synaptic Transmission and Behavioral Deficits.
SHANK3 (also known as PROSAP2) is a postsynaptic scaffolding protein at excitatory synapses in which mutations and deletions have been implicated in patients with idiopathic autism, Phelan-McDermid (aka 22q13 microdeletion) syndrome, and other neuropsychiatric disorders. In this study, we have created a novel mouse model of human autism caused by the insertion of a single guanine nucleotide into exon 21 (Shank3(G)). The resulting frameshift causes a premature STOP codon and loss of major higher molecular weight Shank3 isoforms at the synapse. Shank3(G/G) mice exhibit deficits in hippocampus-dependent spatial learning, impaired motor coordination, altered response to novelty, and sensory processing deficits. At the cellular level, Shank3(G/G) mice also exhibit impaired hippocampal excitatory transmission and plasticity as well as changes in baseline NMDA receptor-mediated synaptic responses. This work identifies clear alterations in synaptic function and behavior in a novel, genetically accurate mouse model of autism mimicking an autism-associated insertion mutation. Furthermore, these findings lay the foundation for future studies aimed to validate and study region-selective and temporally selective genetic reversal studies in the Shank3(G/G) mouse that was engineered with such future experiments in mind.


Ultra-large library docking for discovering new chemotypes.
Despite intense interest in expanding chemical space, libraries containing hundreds-of-millions to billions of diverse molecules have remained inaccessible. Here we investigate structure-based docking of 170 million make-on-demand compounds from 130 well-characterized reactions. The resulting library is diverse, representing over 10.7 million scaffolds that are otherwise unavailable. For each compound in the library, docking against AmpC β-lactamase (AmpC) and the D4 dopamine receptor were simulated. From the top-ranking molecules, 44 and 549 compounds were synthesized and tested for interactions with AmpC and the D4 dopamine receptor, respectively. We found a phenolate inhibitor of AmpC, which revealed a group of inhibitors without known precedent. This molecule was optimized to 77 nM, which places it among the most potent non-covalent AmpC inhibitors known. Crystal structures of this and other AmpC inhibitors confirmed the docking predictions. Against the D4 dopamine receptor, hit rates fell almost monotonically with docking score, and a hit-rate versus score curve predicted that the library contained 453,000 ligands for the D4 dopamine receptor. Of 81 new chemotypes discovered, 30 showed submicromolar activity, including a 180-pM subtype-selective agonist of the D4 dopamine receptor.


Migration of motor pool activity in the spinal cord reflects body mechanics in human locomotion.
During the evolution of bipedal modes of locomotion, a sequential rostrocaudal activation of trunk muscles due to the undulatory body movements was replaced by more complex and discrete bursts of activity. Nevertheless, the capacity for segmental rhythmogenesis and the rostrocaudal propagation of spinal cord activity has been conserved. In humans, motoneurons of different muscles are arranged in columns, with a specific grouping of muscles at any given segmental level. The muscle patterns of locomotor activity and the biomechanics of the body center of mass have been studied extensively, but their interrelationship remains poorly understood. Here we mapped the electromyographic activity recorded from 30 bilateral leg muscles onto the spinal cord in approximate rostrocaudal locations of the motoneuron pools during walking and running in humans. We found that the rostrocaudal displacements of the center of bilateral motoneuron activity mirrored the changes in the energy due to the center-of-body mass motion. The results suggest that biomechanical mechanisms of locomotion, such as the inverted pendulum in walking and the pogo-stick bouncing in running, may be tightly correlated with specific modes of progression of motor pool activity rostrocaudally in the spinal cord.


Field Evaluation of N95 Filtering Facepiece Respirators on Construction Jobsites for Protection against Airborne Ultrafine Particles.
Exposure to high concentrations of airborne ultrafine particles in construction jobsites may play an important role in the adverse health effects among construction workers, therefore adequate respiratory protection is required. The performance of particulate respirators has never been evaluated in field conditions against ultrafine particles on construction jobsites. In this study, respiratory protection levels against ultrafine particles of different size ranges were assessed during three common construction related jobs using a manikin-based set-up at 85 L/min air flow rate. Two NanoScan SMPS nanoparticle counters were utilized for measuring ultrafine particles in two sampling lines of the test filtering facepiece respirator-one from inside the respirator and one from outside the respirator. Particle size distributions were characterized using the NanoScan data collected from outside of the respirator. Two models of N95 respirators were tested-foldable and pleated. Collected data indicate that penetration of all categories of ultrafine particles can exceed 5% and smaller ultrafine particles of <36.5 nm size generally penetrated least. Foldable N95 filtering facepiece respirators were found to be less efficient than pleated N95 respirators in filtering nanoparticles mostly at the soil moving site and the wooden building frameworks construction site. Upon charge neutralization by isopropanol treatment, the ultrafine particles of larger sizes penetrated more compared to particles of smaller sizes. Our findings, therefore, indicate that N95 filtering facepiece respirators may not provide desirable 95% protection for most categories of ultrafine particles and generally, 95% protection is achievable for smaller particles of 11.5 to 20.5 nm sizes. We also conclude that foldable N95 respirators are less efficient than pleated N95 respirators in filtering ultrafine particles, mostly in the soil moving site and the wooden building framework construction site.


Understanding Municipal Officials' Involvement in Transportation Policies Supportive of Walking and Bicycling.
Local transportation policies can impact the built environment and physical activity. Municipal officials play a critical role in transportation policy and planning decisions, yet little is known about what influences their involvement. To describe municipal officials' involvement in transportation policies that were supportive of walking and bicycling and to examine individual- and job-related predictors of involvement in transportation policies among municipal officials. A cross-sectional survey was administered online from June to July 2012 to municipal officials in 83 urban areas with a population of 50 000 or more residents across 8 states. A total of 461 municipal officials from public health, planning, transportation, public works, community and economic development, parks and recreation, city management, and municipal legislatures responded to the survey. Participation in the development, adoption, or implementation of a municipal transportation policy supportive of walking or bicycling. Multivariate logistic regression analyses, conducted in September 2013, revealed that perceived importance of economic development and traffic congestion was positively associated with involvement in a municipal transportation policy (odds ratio [OR] = 1.32, 95% confidence interval [CI] = 1.02-1.70; OR = 1.59, 95% CI = 1.26-2.01, respectively). Higher perceived resident support of local government to address economic development was associated with an increased likelihood of participation in a transportation policy (OR = 1.70, 95% CI = 1.24-2.32). Respondents who perceived lack of collaboration as a barrier were less likely to be involved in a transportation policy (OR = 0.78, 95% CI = 0.63-0.97). Municipal officials who lived in the city or town in which they worked were significantly more likely to be involved in a transportation policy (OR = 1.83, 95% CI = 1.05-3.17). Involvement in a local transportation policy by a municipal official was associated with greater perceived importance of economic development and traffic congestion in job responsibilities, greater perceived resident support of local government to address economic development, and residence of the municipal official. Lack of collaboration represented a barrier to local transportation policy participation.


Multiple internal reflections in the cochlea and their effect on DPOAE fine structure.
In recent years, evidence has accumulated in support of a two-source model of distortion product otoacoustic emissions (DPOAEs). According to such models DPOAEs recorded in the ear canal are associated with two separate sources of cochlear origin. It is the interference between the contributions from the two sources that gives rise to the DPOAE fine structure (a pseudoperiodic change in DPOAE level or group delay with frequency). Multiple internal reflections between the base of the cochlea (oval window) and the DP tonotopic place can add additional significant components for certain stimulus conditions and thus modify the DPOAE fine structure. DPOAEs, at frequency increments between 4 and 8 Hz, were recorded at fixed f2/f1 ratios of 1.053, 1.065, 1.08, 1.11, 1.14, 1.18, 1.22, 1.26, 1.30, 1.32, 1.34, and 1.36 from four subjects. The resulting patterns of DPOAE amplitude and group delay (the negative of the slope of phase) revealed several previously unreported patterns in addition to the commonly reported log sine variation with frequency. These observed "exotic" patterns are predicted in computational simulations when multiple internal reflections are included. An inverse FFT algorithm was used to convert DPOAE data from the frequency to the "time" domain. Comparison of data in the time and frequency domains confirmed the occurrence of these "exotic" patterns in conjunction with the presence of multiple internal reflections. Multiple internal reflections were observed more commonly for high primary ratios (f2/f1 > or = 1.3). These results indicate that a full interpretation of the DPOAE level and phase (group delay) must include not only the two generation sources, but also multiple internal reflections.


Isolation of pathogenic aerobic bacteria from the blood of septicaemic neonatal calves and the susceptibility of isolates to various antibiotics.
An automated blood culture system (BACTEC 9240) was used for the isolation of aerobic bacteria from the blood of septicaemic neonatal calves. Blood samples were collected from 90 clinically septicaemic and 20 healthy neonatal calves and inoculated into blood culture bottles. There were 89 significant isolates from 90 positive blood cultures using the BACTEC system. Escherichia coli was the most common pathogen detected accounting for 56 (63%) out of 89 isolates. The other pathogens were beta-haemolytic streptococci (15.7%), Staphylococcus aureus (10.1%), Klebsiella sp. (5.6%) and Corynebacterium sp. (5.6%). All isolates showed a susceptibility rate of 100% to enrofloxacin, cefepim, cefoperazone/sulbactam, imipenem and meropenem while some of them were ranged from 75 to 91.7% susceptible to amoxicillin/clavulanic acid, ampicillin/sulbactam, gentamicin and cephalosporins.


Long-term Outcome After Survival of a Cardiac Arrest: A Prospective Longitudinal Cohort Study.
A cardiac arrest can lead to hypoxic brain injury, which can affect all levels of functioning. To investigate 1-year outcome and the pattern of recovery after surviving a cardiac arrest. This was a multicenter, prospective longitudinal cohort study with 1 year of follow-up (measurements 2 weeks, 3 months, 1 year). On function level, physical/cardiac function (New York Heart Association Classification), cognition (Cognitive Log [Cog-log], Cognitive Failures Questionnaire), emotional functioning (Hospital Anxiety and Depression Scale, Impact of Event Scale), and fatigue (Fatigue Severity Scale) were assessed. In addition, level of activities (Frenchay Activities Index, FAI), participation (Community Integration Questionnaire [CIQ] and return to work), and quality of life (EuroQol 5D, EuroQol Visual Analogue Scale, SF-36, Quality of Life after Brain Injury) were measured. In this cohort, 141 cardiac arrest survivors were included. At 1 year, 14 (13%) survivors scored below cutoff on the Cog-log. Both anxiety and depression were present in 16 (15%) survivors, 29 (28%) experienced posttraumatic stress symptoms and 55 (52%), severe fatigue. Scores on the FAI and the CIQ were, on average, respectively 96% and 92% of the prearrest scores. Of those previously working, 41 (72%) had returned to work. Most recovery of cognitive function and quality of life occurred within the first 3 months, with further improvement on some domains of quality of life up to 12 months. Overall, long-term outcome in terms of activities, participation, and quality of life after cardiac arrest is reassuring. Nevertheless, fatigue is common; problems with cognition and emotions occur; and return to work can be at risk.