Structural analysis of heavy oil fractions after hydrodenitrogenation by high-resolution tandem mass spectrometry and ion mobility spectrometry.
Heavy petroleum fractions such as vacuum gas oils (VGOs) are structurally and compositionally highly complex mixtures. Nitrogen species, which have a significant impact on the subsequent refining processes, are generally removed by the hydrodenitrogenation (HDN) catalytic process. The purpose of this study was to identify and characterize compounds that are refractory to the HDN process. This may allow for the examination of the effectiveness of a vacuum distillate hydrotreatment catalytic bed in removing nitrogen-containing compounds before the cracking step. Three different VGO fractions of the same oil before and after HDN processes were analysed in ESI(+) mode by FTICR mass spectrometry and ion mobility spectrometry-mass spectrometry (IMS-MS), in particular compounds containing basic nitrogen, such as quinoline and isoquinoline. Ultra-high-resolution FTICR mass spectrometry provides a sufficiently high mass resolution power to resolve different compounds and attribute a unique molecular formula to each ion. Information on the isomeric content was obtained by use of tandem mass spectrometry (MS/MS) and IMS-MS. The evolution of the fragmentation of the N1 class of compounds as a function of collision energy allowed for the identification of the molecular nucleus raw formula. From the IMS-MS experiments, it clearly appeared that, based on the IMS peak width, a lower isomeric dispersity was obtained after the HDN process and, based on the drift time and collision cross section determination, species presenting longer alkyl branches are the molecules most refractory to the HDN process.


The composition and dynamics of cell-substratum adhesions in locomoting fish keratocytes.
Close contacts are the predominant type of cell-substratum adhesion in rapidly moving cells yet little is known about their composition and dynamics. To address these issues we have attempted to identify the molecular components of close contacts formed in rapidly moving fish epithelial keratocytes. In addition we have utilized the simple shape of keratocytes to explore the relationship between close contact formation and rapid locomotion. Beta1 integrin and talin molecules were found to be localized within a narrow rim of very close contact along the leading edge. These molecules together with vinculin were also found within small foci distributed evenly throughout the lamella, corresponding to regions of variable close contact. Alpha-actinin was found in foci within older, more posteriorly located regions of the lamella and along stress fibers. In addition to close contacts, small focal adhesion-like structures which stained positively for all antibodies tested were found at the tips of stress fibers within retracting cell margins. Interference reflection and total internal reflection microscopy of moving keratocytes showed cell-substratum contacts to be organized into distinct patterns that appear to move forwards, in concert with the leading edge. A feature common to all cells is a rim of very close contact at the leading edge. This region is specialized for the formation of new cell-substratum adhesions and is the site where patterns of close contact are generated. We have found that cell locomotion is most rapid when a uniform contact pattern is present but cell speed is progressively reduced as the contact pattern becomes more irregular. Furthermore, the local rate of lamellar extension is most rapid when underlain by regions of intermediate closeness to the substratum, but is reduced or ceases if the underlying contact is either very close or more distant, respectively. Our results suggest that close contacts and focal adhesions are related structures formed from a common hierarchy of molecular interactions. In addition the relationship between close contact formation and lamellar extension indicates a direct coupling between these two processes at the leading edge. Furthermore, we can explain the dynamic behaviour of close contacts in terms of the relative rates of trapping and release of component molecules that is initiated at the leading edge. We suggest that regulation of the molecular dynamics involved in leading edge specialization determines both the pattern of cell-substratum contacts and the net rate of actin filament assembly.


Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly.
The Endosomal Sorting Complexes Required for Transport III (ESCRT-III) proteins are critical for cellular membrane scission processes with topologies inverted relative to clathrin-mediated endocytosis. Some viruses appropriate ESCRT-IIIs for their release. By imaging single assembling viral-like particles of HIV-1, we observed that ESCRT-IIIs and the ATPase VPS4 arrive after most of the virion membrane is bent, linger for tens of seconds, and depart ~20 s before scission. These observations suggest that ESCRT-IIIs are recruited by a combination of membrane curvature and the late domains of the HIV-1 Gag protein. ESCRT-IIIs may pull the neck into a narrower form but must leave to allow scission. If scission does not occur within minutes of ESCRT departure, ESCRT-IIIs and VPS4 are recruited again. This mechanistic insight is likely relevant for other ESCRT-dependent scission processes including cell division, endosome tubulation, multivesicular body and nuclear envelope formation, and secretion of exosomes and ectosomes.


Multiple myocardial hamartomas causing ventricular tachycardia in young children: combined surgical modification and medical treatment.
To describe two cases of incessant ventricular tachycardia caused by multiple myocardial hamartomas. Two infants, 13 and 14 months old, who had had multiple episodes of symptomatic tachycardia were referred to our institution. Incessant ventricular tachycardia was diagnosed. Initially, the patients received pharmacologic therapy. The recurrent tachycardia resulted in notable hemodynamic instability. No structural abnormalities were detected on the echocardiograms. In one patient, an electrophysiologic study revealed that the site of ventricular ectopic beats was in the anterolateral wall of the left ventricle, midway between the apex and the base. In the other patient, a preoperative electrophysiologic study was not undertaken because of the inability to obtain central venous access. Diffuse hamartomas were found throughout the ventricular myocardium in both patients. Surgical resection and cryoablation of the lesions in combination with medical therapy helped control the tachyarrhythmia. At 7 and 17 months postoperatively, the patients were in normal sinus rhythm and were receiving medication. In young children who have incessant ventricular tachycardia but no lesion evident on echocardiography, angiography, or other imaging modalities, a myocardial tumor should be suspected, and pharmacologic therapy should be instituted. If the medical regimen fails, surgical intervention should be undertaken, directed at areas localized by inspection and by preoperative and intraoperative electrophysiologic studies.


Pituitary Adenylate Cyclase-Activating Polypeptide Disrupts Motivation, Social Interaction, and Attention in Male Sprague Dawley Rats.
Severe or prolonged stress can trigger psychiatric illnesses including mood and anxiety disorders. Recent work indicates that pituitary adenylate cyclase-activating polypeptide (PACAP) plays an important role in regulating stress effects. In rodents, exogenous PACAP administration can produce persistent elevations in the acoustic startle response, which may reflect anxiety-like signs including hypervigilance. We investigated whether PACAP causes acute or persistent alterations in behaviors that reflect other core features of mood and anxiety disorders (motivation, social interaction, and attention). Using male Sprague Dawley rats, we examined if PACAP (.25-1.0 µg, intracerebroventricular infusion) affects motivation as measured in the intracranial self-stimulation test. We also examined if PACAP alters interactions with a conspecific in the social interaction test. Finally, we examined if PACAP affects performance in the 5-choice serial reaction time task, which quantifies attention and error processing. Dose-dependent disruptions in motivation, social interaction, and attention were produced by PACAP, as reflected by increases in reward thresholds, decreases in social behaviors, and decreases in correct responses and alterations in posterror accuracy. Behavior normalized quickly in the intracranial self-stimulation and 5-choice serial reaction time task tests but remained dysregulated in the social interaction test. Effects on attention were attenuated by the corticotropin-releasing factor receptor-1 antagonist antalarmin but not the κ opioid receptor antagonist JDTic. Our findings suggest that PACAP affects numerous domains often dysregulated in mood and anxiety disorders, but that individual signs depend on brain substrates that are at least partially independent. This work may help to devise therapeutics that mitigate specific signs of these disorders.


Pleiotropic role of the RNA chaperone protein Hfq in the human pathogen Clostridium difficile.
Clostridium difficile is an emergent human pathogen and the most common cause of nosocomial diarrhea. Our recent data strongly suggest the importance of RNA-based mechanisms for the control of gene expression in C. difficile. In an effort to understand the function of the RNA chaperone protein Hfq, we constructed and characterized an Hfq-depleted strain in C. difficile. Hfq depletion led to a growth defect, morphological changes, an increased sensitivity to stresses, and a better ability to sporulate and to form biofilms. The transcriptome analysis revealed pleiotropic effects of Hfq depletion on gene expression in C. difficile, including genes encoding proteins involved in sporulation, stress response, metabolic pathways, cell wall-associated proteins, transporters, and transcriptional regulators and genes of unknown function. Remarkably, a great number of genes of the regulon dependent on sporulation-specific sigma factor, SigK, were upregulated in the Hfq-depleted strain. The altered accumulation of several sRNAs and interaction of Hfq with selected sRNAs suggest potential involvement of Hfq in these regulatory RNA functions. Altogether, these results suggest the pleiotropic role of Hfq protein in C. difficile physiology, including processes important for the C. difficile infection cycle, and expand our knowledge of Hfq-dependent regulation in Gram-positive bacteria.


Identification of Salmonella enterica serovar Typhimurium using specific PCR primers obtained by comparative genomics in Salmonella serovars.
Salmonella enterica serovar Typhimurium is a major foodborne pathogen throughout the world. Until now, the specific target genes for the detection and identification of serovar Typhimurium have not been developed. To determine the specific probes for serovar Typhimurium, the genes of serovar Typhimurium LT2 that were expected to be unique were selected with the BLAST (Basic Local Alignment Search Tool) program within GenBank. The selected genes were compared with 11 genomic sequences of various Salmonella serovars by BLAST. Of these selected genes, 10 were expected to be specific to serovar Typhimurium and were not related to virulence factor genes of Salmonella pathogenicity island or to genes of the O and H antigens of Salmonella. Primers for the 10 selected genes were constructed, and PCRs were evaluated with various genomic DNAs of Salmonella and non-Salmonella strains for the specific identification of Salmonella serovar Typhimurium. Among all the primer sets for the 10 genes, STM4497 showed the highest degree of specificity to serovar Typhimurium. In this study, a specific primer set for Salmonella serovar Typhimurium was developed on the basis of the comparison of genomic sequences between Salmonella serovars and was validated with PCR. This method of comparative genomics to select target genes or sequences can be applied to the specific detection of microorganisms.


Tuning the supramolecular isomerism of MOF-74 by controlling the synthesis conditions.
Supramolecular isomerism of metal-organic frameworks (MOFs) is known for several MOF structures, having direct implications on the properties of these materials. Although the synthesis of MOF isomers is mainly serendipitous in nature, achieving controlled formation of a target framework is highly relevant for practical applications. This work discusses the influence of additives and synthesis conditions on the formation of porous isomers containing Zn2+ as nodes and 2,5-dihydroxy-1,4-benzenedicarboxylate (dobdc4-) as a linker. Using solvent mixtures containing strongly coordinated molecules, e.g. N,N'-dimethylformamide (DMF) and N-methylpyrrolidone (NMP), facilitates the formation of porous structures of type [Zn2(dobdc)(S)x]·yS (S = DMF, NMP) which are built from dinuclear Zn2(O)2(CO2)3 secondary building units (SBUs) consisting of two different edge-sharing polyhedra with the Zn2+ ions in a unsaturated coordinative environment. In the presence of water, the Zn2+ dimers are converted to one-dimensional infinite Zn2+ chains, in which the number of Zn2+-linker bonds increases, therefore giving a hydrolytically more stable coordination environment. The full characterization of the isomers as well as their conversion to the most stable isomer is presented.


Protective alleles and modifier variants in human health and disease.
The combination of next-generation sequencing technologies and high-throughput genotyping platforms has revolutionized the pursuit of genetic variants that contribute towards disease. Furthermore, these technologies have provided invaluable insight into the genetic factors that prevent individuals from developing disease. Exploiting the evolutionary mechanisms that were designed by nature to help prevent disease is an attractive line of enquiry. Such efforts have the potential to generate a therapeutic target roadmap and rejuvenate the current drug-discovery pathway. By delineating the genomic factors that are protective against disease, there is potential to derive highly effective, genomically anchored medicines that assist in maintaining health.


Effect of prenatal glucocorticoids and thyroid hormones on developmental plasticity of mitochondrial aerobic metabolism, growth and survival: an experimental test in wild great tits.
Developmental plasticity is partly mediated by transgenerational effects, including those mediated by the maternal endocrine system. Glucocorticoid and thyroid hormones may play central roles in developmental programming through their action on metabolism and growth. However, the mechanisms by which they affect growth and development remain understudied. One hypothesis is that maternal hormones directly affect the production and availability of energy-carrying molecules (e.g. ATP) by their action on mitochondrial function. To test this hypothesis, we experimentally increased glucocorticoid and thyroid hormones in wild great tit eggs (Parus major) to investigate their impact on offspring mitochondrial aerobic metabolism (measured in blood cells), and subsequent growth and survival. We show that prenatal glucocorticoid supplementation affected offspring cellular aerobic metabolism by decreasing mitochondrial density, maximal mitochondrial respiration and oxidative phosphorylation, while increasing the proportion of the maximum capacity being used under endogenous conditions. Prenatal glucocorticoid supplementation only had mild effects on offspring body mass, size and condition during the rearing period, but led to a sex-specific (females only) decrease in body mass a few months after fledging. Contrary to our expectations, thyroid hormone supplementation did not affect offspring growth or mitochondrial metabolism. Recapture probability as juveniles or adults was not significantly affected by prenatal hormonal treatment. Our results demonstrate that prenatal glucocorticoids can affect post-natal mitochondrial density and aerobic metabolism. The weak effects on growth and apparent survival suggest that nestlings were mostly able to compensate for the transient decrease in mitochondrial aerobic metabolism induced by prenatal glucocorticoids.


Clobetasol propionate 0.05% spray for the management of moderate-to-severe plaque psoriasis of the scalp: results from a randomized controlled trial.
Clobetasol propionate 0.05% spray is available for treating moderate-to-severe plaque psoriasis; however, there is limited information with plaque psoriasis of the scalp. Evaluate the efficacy, safety, and quality-of-life impact of clobetasol propionate 0.05% spray in patients with moderate to severe plaque psoriasis of the scalp. Multicenter, randomized, double-blind, vehicle-controlled study involving 81 men and women with moderate-to-severe (Global Severity Score [GSS] = 3 or 4) plaque psoriasis of the scalp. Eligible patients were treated with clobetasol propionate 0.05% spray or vehicle spray, which was applied twice daily for up to four weeks. The primary efficacy end point was the GSS of psoriasis of the scalp after four weeks. Safety assessments included local tolerability, presence of Cushing's syndrome, and adverse events. At the end of treatment, 85 percent (35/41) of patients in the clobetasol propionate 0.05% spray group achieved success (GSS clear or almost clear), compared with 13 percent (5/40) in the vehicle spray group (P is less than .001). The proportion of patients treated with clobetasol propionate 0.05% spray who achieved a rating of clear (GSS = 0) after two weeks and at the end of treatment was 12 percent and 51 percent, respectively. Clobetasol propionate 0.05% spray was well tolerated, and there were no serious adverse events or reported cases of folliculitis or Cushing's syndrome. Treatment with clobetasol propionate 0.05% spray for up to four weeks is effective and well tolerated for moderate-to-severe plaque psoriasis of the scalp.


LFA-1 activates focal adhesion kinases FAK1/PYK2 to generate LAT-GRB2-SKAP1 complexes that terminate T-cell conjugate formation.
Lymphocyte function-associated antigen 1 (LFA-1) affinity and avidity changes have been assumed to mediate adhesion to intercellular adhesion molecule-1 for T-cell conjugation to dendritic cells (DC). Although the T-cell receptor (TCR) and LFA-1 can generate intracellular signals, the immune cell adaptor protein linker for the activation of T cells (LAT) couples the TCR to downstream events. Here, we show that LFA-1 can mediate both adhesion and de-adhesion, dependent on receptor clustering. Although increased affinity mediates adhesion, LFA-1 cross-linking induced the association and activation of the protein-tyrosine kinases FAK1/PYK1 that phosphorylated LAT selectively on a single Y-171 site for the binding to adaptor complex GRB-2-SKAP1. LAT-GRB2-SKAP1 complexes were distinct from canonical LAT-GADs-SLP-76 complexes. LFA-1 cross-linking increased the presence of LAT-GRB2-SKAP1 complexes relative to LAT-GADs-SLP-76 complexes. LFA-1-FAK1 decreased T-cell-dendritic cell (DC) dwell times dependent on LAT-Y171, leading to reduced DO11.10 T cell binding to DCs and proliferation to OVA peptide. Overall, our findings outline a new model for LFA-1 in which the integrin can mediate both adhesion and de-adhesion events dependent on receptor cross-linking.


Neural correlates of abnormal sensory discrimination in laryngeal dystonia.
Aberrant sensory processing plays a fundamental role in the pathophysiology of dystonia; however, its underpinning neural mechanisms in relation to dystonia phenotype and genotype remain unclear. We examined temporal and spatial discrimination thresholds in patients with isolated laryngeal form of dystonia (LD), who exhibited different clinical phenotypes (adductor vs. abductor forms) and potentially different genotypes (sporadic vs. familial forms). We correlated our behavioral findings with the brain gray matter volume and functional activity during resting and symptomatic speech production. We found that temporal but not spatial discrimination was significantly altered across all forms of LD, with higher frequency of abnormalities seen in familial than sporadic patients. Common neural correlates of abnormal temporal discrimination across all forms were found with structural and functional changes in the middle frontal and primary somatosensory cortices. In addition, patients with familial LD had greater cerebellar involvement in processing of altered temporal discrimination, whereas sporadic LD patients had greater recruitment of the putamen and sensorimotor cortex. Based on the clinical phenotype, adductor form-specific correlations between abnormal discrimination and brain changes were found in the frontal cortex, whereas abductor form-specific correlations were observed in the cerebellum and putamen. Our behavioral and neuroimaging findings outline the relationship of abnormal sensory discrimination with the phenotype and genotype of isolated LD, suggesting the presence of potentially divergent pathophysiological pathways underlying different manifestations of this disorder.


NSABP B-47/NRG Oncology Phase III Randomized Trial Comparing Adjuvant Chemotherapy With or Without Trastuzumab in High-Risk Invasive Breast Cancer Negative for HER2 by FISH and With IHC 1+ or 2.
Adjuvant trastuzumab reduces invasive breast cancer (IBC) recurrence and risk for death in patients with HER2-amplified or overexpressing IBC. A subset of patients in the landmark trastuzumab adjuvant trials who originally tested HER2-positive but were HER2-negative by central HER2 testing appeared to possibly benefit from trastuzumab. The objective for the NSABP B-47 trial was to determine whether the addition of trastuzumab to adjuvant chemotherapy (CRx) would improve invasive disease-free survival (IDFS) in patients with HER2-negative breast cancer. A total of 3,270 women with high-risk primary IBC were randomly assigned to CRx with or without 1 year of trastuzumab. Eligibility criteria included immunohistochemistry (IHC) score 1+ or 2+ with fluorescence in situ hybridization ratio (FISH) < 2.0 or, if ratio was not performed, HER2 gene copy number < 4.0. CRx was either docetaxel plus cyclophosphamide or doxorubicin and cyclophosphamide followed by weekly paclitaxel for 12 weeks. At a median follow-up of 46 months, the addition of trastuzumab to CRx did not improve IDFS (5-year IDFS: 89.8% with CRx plus trastuzumab [CRxT] v 89.2% with CRx alone; hazard ratio [HR], 0.98; 95% CI, 0.76 to 1.25; P = .85). These findings did not differ by level of HER2 IHC expression, lymph node involvement, or hormone-receptor status. For distant recurrence-free interval, 5-year estimates were 92.7% with CRxT compared with 93.6% for CRx alone (HR, 1.10; 95% CI, 0.81 to 1.50; P = .55) and for overall survival (OS) were 94.8% with CRxT and 96.3% in CRx alone (HR, 1.33; 95% CI, 0.90 to 1.95; P = .15). There were no unexpected toxicities from the addition of trastuzumab to CRx. The addition of trastuzumab to CRx did not improve IDFS, distant recurrence-free interval, or OS in women with non-HER2-overexpressing IBC. Trastuzumab does not benefit women without IHC 3+ or FISH ratio-amplified breast cancer.


The heliospheric magnetic field over the South polar region of the sun.
Magnetic field measurements from the Ulysses space mission overthe south polar regions of the sun showed that the structure and properties of the three-dimensional heliosphere were determined by the fast solar wind flow and magnetic fields from the large coronal holes in the polar regions of the sun. This conclusion applies at the current, minimum phase of the 11-year solar activity cycle. Unexpectedly, the radial component of the magnetic field was independent of latitude. The high-latitude magnetic field deviated significantly from the expected Parker geometry, probably because of large amplitude transverse fluctuations. Low-frequency fluctuations had a high level of variance. The rate of occurrence of discontinuities also increased significantly at high latitudes.


Islet Transplantation in the UK.
This commentary outlines the development and current status of the UK Islet Transplant Programme in the UK. The author makes the case that it is now time for similar fully funded beta-cell programmes to be made available in many other countries as well.


Cell-specific cardiac electrophysiology models.
The traditional cardiac model-building paradigm involves constructing a composite model using data collected from many cells. Equations are derived for each relevant cellular component (e.g., ion channel, exchanger) independently. After the equations for all components are combined to form the composite model, a subset of parameters is tuned, often arbitrarily and by hand, until the model output matches a target objective, such as an action potential. Unfortunately, such models often fail to accurately simulate behavior that is dynamically dissimilar (e.g., arrhythmia) to the simple target objective to which the model was fit. In this study, we develop a new approach in which data are collected via a series of complex electrophysiology protocols from single cardiac myocytes and then used to tune model parameters via a parallel fitting method known as a genetic algorithm (GA). The dynamical complexity of the electrophysiological data, which can only be fit by an automated method such as a GA, leads to more accurately parameterized models that can simulate rich cardiac dynamics. The feasibility of the method is first validated computationally, after which it is used to develop models of isolated guinea pig ventricular myocytes that simulate the electrophysiological dynamics significantly better than does a standard guinea pig model. In addition to improving model fidelity generally, this approach can be used to generate a cell-specific model. By so doing, the approach may be useful in applications ranging from studying the implications of cell-to-cell variability to the prediction of intersubject differences in response to pharmacological treatment.


Testimony by otolaryngologists in defense of tobacco companies 2009-2014.
To examine expert testimony offered by otolaryngologists in defense of the tobacco industry and to assess whether opinions rendered were congruent with evidence in the scientific literature. Data sources include publically available expert witness depositions and trial testimony of board-certified otolaryngologists employed by the tobacco industry in defense of lawsuits brought by smokers suffering from head and neck cancer. The cases, adjudicated in Florida between 2009 and 2014, focused on whether smoking caused the plaintiff's cancer. The study includes nine legal cases of upper aerodigestive tract cancer involving six otolaryngologists serving as expert witnesses for the tobacco industry. Cancer sites included larynx (5), esophagus (2), mouth (1), and lung (1). Five of the six otolaryngologists consistently, over multiple cases, offered opinions that smoking did not cause the plaintiff's cancer. By highlighting an exhaustive list of potential risk factors, such as human papillomavirus (HPV), alcohol, asbestos, diesel fumes, salted fish, mouthwash, and even urban living, they created doubt in the minds of the jurors as to the role of smoking in the plaintiff's cancer. Evidence shows that this testimony, which was remarkably similar across cases, was part of a defense strategy shaped by tobacco's law firms. A small group of otolaryngologists regularly serve as experts on behalf of the tobacco industry. Examination of their opinions in relation to the scientific literature reveals a systematic bias in interpreting the data relating to the role played by smoking in head and neck cancer causation. N/A.


The CRASH trial protocol (Corticosteroid randomisation after significant head injury) [ISRCTN74459797].
BACKGROUND: Worldwide, millions of people are treated each year for significant head injury. A substantial proportion die, and many more are disabled. If short term corticosteroid infusion could be reliably shown to reduce these risks by just a few percent then this might affect the treatment of a few hundred thousand patients a year, protecting thousands from death or long term disability. STUDY DESIGN: CRASH is a large simple, placebo-controlled trial of the effects of a 48-hour infusion of corticosteroids on death and on neurological disability, among adults with head injury and some impairment of consciousness. Head injured patients with impaired consciousness who are judged to be 16 years or older are eligible if the responsible doctor is, for any reason, substantially uncertain whether or not to use corticosteroids. ORGANISATION: The CRASH trial will determine reliably the effects on death and disability of a short corticosteroid infusion following significant head injury. To detect or refute improvements of only a few percent in outcome, many thousands of acute head injury patients must be randomised between control and steroid infusions. Such large numbers will be possible only if hundreds of doctors and nurses can collaborate in the participating emergency departments. Since they are busy, and working in emergency situations, the trial involves them in almost no extra work: no special investigations or changes to usual management are required, and data collection is absolutely minimal. The trial is on-going and new collaborators are welcome. Further information about the trial is available at http://www.crash.lshtm.ac.uk


The effects of whole mushrooms during inflammation.
Consumption of edible mushrooms has been suggested to improve health. A number of isolated mushroom constituents have been shown to modulate immunity. Five commonly consumed edible mushrooms were tested to determine whether whole mushrooms stimulate the immune system in vitro and in vivo. The white button (WB) extracts readily stimulated macrophage production of TNF-alpha. The crimini, maitake, oyster and shiitake extracts also stimulated TNF-alpha production in macrophage but the levels were lower than from WB stimulation. Primary cultures of murine macrophage and ovalbumin (OVA) specific T cells showed that whole mushroom extracts alone had no effect on cytokine production but co-stimulation with either lipopolysaccharide or OVA (respectively) induced TNF-alpha, IFN-gamma, and IL-1beta while decreasing IL-10. Feeding mice diets that contained 2% WB mushrooms for 4 weeks had no effect on the ex vivo immune responsiveness or associated toxicity (changes in weight or pathology of liver, kidney and gastrointestinal tract). Dextran sodium sulfate (DSS) stimulation of mice that were fed 1% WB mushrooms were protected from DSS induced weight loss. In addition, 2% WB feeding protected the mice from transient DSS induced colonic injury. The TNF-alpha response in the colon and serum of the DSS challenged and 2% WB fed mice was higher than controls. The data support a model whereby edible mushrooms regulate immunity in vitro. The in vivo effects of edible mushrooms required a challenge with DSS to detect small changes in TNF-alpha and transient protection from colonic injury. There are modest effects of in vivo consumption of edible mushrooms on induced inflammatory responses. The result is not surprising since it would certainly be harmful to strongly induce or suppress immune function following ingestion of a commonly consumed food.


Effects of occupational exposure to mercury vapour on the central nervous system.
Possible effects of mercury on the central nervous system (CNS) were examined in a group of chloralkali workers exposed to mercury (n = 89) and compared with a control group (n = 75), by registration of subjective symptoms, personality changes, forearm tremor, and performance on six computerised psychometric tests in the two groups. The groups were similar in age, education, verbal comprehension, and work tasks. In the chloralkali group, median blood mercury concentration (B-Hg) was 55 nmol/l, serum mercury concentration (S-Hg) 45 nmol/l, and urine mercury concentration (U-Hg) 14.3 nmol/mmol creatinine (25.4 micrograms/g creatinine). Corresponding concentrations in the control group were 15 nmol/l, 4 nmol/l, and 1.1 nmol/mmol creatinine (1.9 micrograms/g creatinine) respectively. The number of self reported symptoms, the scores for tiredness and confusion in the profile of mood states (POMS), and the degree of neuroticism in the Eysenck personality inventory (EPI), were significantly higher in the mercury exposed group compared with the controls. Performance on the psychometric tests and tremor frequency spectra did not differ significantly between the two groups. Dose-response calculations showed weak but statistically significant relations between symptom prevalence and current mercury concentrations in both blood and urine. The performance on three of the psychometric tests was negatively correlated with earlier peak exposures. The findings indicate a slight mercury induced effect on the CNS among the chloralkali workers.


Anorexia Nervosa, Bulimia Nervosa, and Binge Eating Disorder in Midlife and Beyond.
We examined eating disorders in midlife and beyond by comparing frequency of anorexia nervosa (AN), bulimia nervosa (BN), binge eating disorder (BED), and other specified feeding or eating disorder (OSFED) among midlife eating disorder treatment-seeking individuals and younger controls. We also compared demographic and eating disorder-related characteristics across diagnoses and age groups. Participants included 2,118 treatment-seeking adults who self-reported their eating-related symptoms on the Eating Disorder Questionnaire. Results showed that percent of patients with BN was significantly lower whereas percent of patients with BED and OSFED was significantly higher among midlife relative to younger patients. Percent of patients with AN did not differ between midlife and younger patients. Additionally, midlife and younger patients with BED and OSFED differed on several demographic (e.g., marital status) and eating disorder-related characteristics (e.g., BMI, compulsive exercising). This study suggests that BN is less common whereas BED and OSFED are more common among midlife eating disorder treatment-seeking individuals relative to younger controls. In addition, AN and BN present fairly similarly whereas BED and OSFED present fairly differently among midlife patients relative to younger controls. Attention to these differences and similarities is necessary to understand eating disorders in midlife.


Chemiluminescence imaging of Duox2-derived hydrogen peroxide for longitudinal visualization of biological response to viral infection in nasal mucosa.
Rationale: Hydrogen peroxide (H2O2) provides an important mechanism for resisting infectious pathogens within the respiratory tract, and accordingly, the in situ analysis of H2O2 generation in real time provides a valuable tool for assessing immune response. Methods: In this study, we applied a chemiluminescent nanoparticle-based real-time imaging approach to noninvasive evaluation of the Duox2-mediated H2O2 generation after viral infection, and assessed its usefulness for analytical purposes in mouse nasal mucosa. The chemiluminescent nanoprobe employed herein (BioNT) possesses appropriate physicochemical properties, such as high sensitivity and selectivity toward H2O2, no background noise, deliverability to the respiratory tract, and capability of multiple injections to a single animal subject for long-term repetitive imaging. Results: The favorable characteristics of BioNT allowed for a longitudinal study with the same mice to noninvasively evaluate the long-term evolution of endogenous H2O2 in the nasal epithelium after infection with influenza A virus (WS/33/H1N1). We found that nasal epithelial cells by themselves respond to viral infection by generating H2O2, and that the in vivo cumulative H2O2 level in the nasal mucosa peaks at day 3 post-infection. Such in vitro and in vivo temporal behaviors of the endogenous H2O2 generation showed a good correlation with those of Duox2 expression after infection. This correlation could be further confirmed with Duox2-deficient subjects (Duox2-knockdown NHNE cells and Duox2-knockout mutant mice) where no H2O2-induced chemiluminescence was detectable even after viral infection. Importantly, upon knock-down of Duox2 expression, the condition of mice caused by viral infection in the upper airway was significantly aggravated, evidencing the involvement of Duox2 in the immune defense. Conclusion: All these results reveal a critical role of Duox2 in the infection-induced H2O2 production and the H2O2-mediated immune response to infection in the respiratory tract, well elucidating the potential of BioNT as a noninvasive tool for fundamental in vivo studies of infectious diseases.


A conserved GATA motif in a tissue-specific DNase I hypersensitive site of the cardiac alpha-myosin heavy chain gene.
Transgenic analysis has indicated that far upstream regulatory elements of the cardiac alpha-myosin heavy chain (MyHC) gene are required for appropriate transgene expression [Subramaniam, Gulick, Neumann, Knotts and Robbins (1993) J. Biol. Chem. 268, 4331-4336]. In an attempt to identify these as-yet-undefined regulatory elements, we mapped the DNase I hypersensitive sites (DHSs) in the 4 kb upstream region of the hamster cardiac alpha-MyHC gene. When using nuclei isolated from late-gestational and adult heart ventricles, a strong DHS was identified in the -1.9 kb region (alpha-1.9 kb site). It cannot be detected in kidney, liver or cardiofibroblast nuclei. Within this site, we found a conserved GATA-motif that interacts specifically with GATA-binding factors in nuclear extracts of cardiomyocytes at various developmental stages. These data provide further evidence to support the role of GATA factors in the regulation of cardiac alpha-MyHC gene expression.


Distinctive roles of age, sex, and genetics in shaping transcriptional variation of human immune responses to microbial challenges.
The contribution of host genetic and nongenetic factors to immunological differences in humans remains largely undefined. Here, we generated bacterial-, fungal-, and viral-induced immune transcriptional profiles in an age- and sex-balanced cohort of 1,000 healthy individuals and searched for the determinants of immune response variation. We found that age and sex affected the transcriptional response of most immune-related genes, with age effects being more stimulus-specific relative to sex effects, which were largely shared across conditions. Although specific cell populations mediated the effects of age and sex on gene expression, including CD8+ T cells for age and CD4+ T cells and monocytes for sex, we detected a direct effect of these intrinsic factors for the majority of immune genes. The mapping of expression quantitative trait loci (eQTLs) revealed that genetic factors had a stronger effect on immune gene regulation than age and sex, yet they affected a smaller number of genes. Importantly, we identified numerous genetic variants that manifested their regulatory effects exclusively on immune stimulation, including a Candida albicans-specific master regulator at the CR1 locus. These response eQTLs were enriched in disease-associated variants, particularly for autoimmune and inflammatory disorders, indicating that differences in disease risk may result from regulatory variants exerting their effects only in the presence of immune stress. Together, this study quantifies the respective effects of age, sex, genetics, and cellular heterogeneity on the interindividual variability of immune responses and constitutes a valuable resource for further exploration in the context of different infection risks or disease outcomes.


A new prognostic model for hepatocellular carcinoma recurrence after curative hepatectomy.
We previously reported the effectiveness of the product of tumor number and size (NxS factor) for the prognosis of hepatocellular carcinoma (HCC) in patients following hepatectomy. The present study aimed to propose a new score based on the NxS factor to predict HCC recurrence following hepatectomy. A total of 406 patients who underwent hepatectomy for HCC at Osaka University Graduate School of Medicine were retrospectively analyzed to develop the new score. Among clinicopathological factors, including the NxS factor, the marker subset that achieved the best performance for prediction of early recurrence was assessed, and a prognostic model for HCC recurrence after curative hepatectomy (REACH) was developed. As the validation set, 425 patients who underwent hepatectomy for HCC at Yamaguchi University Graduate School of Medicine and Shimonoseki Medical Center were analyzed, and the prognostic ability of the REACH score was compared with that of well-known staging systems. Following analysis, the REACH score was constructed using six covariates (NxS factor, microscopic hepatic vein invasion, differentiation, serum albumin, platelet count and indocyanine green retention rate at 15 min). In the validation set, the REACH score predicted early recurrence in 73 of 81 samples, with a sensitivity of 89% and a specificity of 58%. The area under the curve (AUC) of the receiver operating characteristic curve of the REACH score was 0.78 and 0.74, respectively, for 1- and 2-year recurrence after hepatectomy; each AUC was higher than that of any of the other staging systems. Survival analysis indicated the REACH score had the best predictive value in disease-free and overall survival. The present findings demonstrated that the REACH score may be used to classify patients with HCC into high- and low-risk of recurrence, and to predict subsequent survival following hepatic resection.


The Predictive Role of Abdominal Fat Parameters and Stone Density on SWL Outcomes.
Our aim was to detect the role of radiological abdominal fat parameters by tomography and stone density by plain X-ray on extracorporeal Shock Wave Lithotripsy (SWL) stone-free rate. The patients who had undergone SWL for a single opaque renal stone < 2 cm in diameter and proximal ureteric stone < 1 cm in diameter were collected retrospectively. The characteristics of patients and stones were recorded. The stone attitude, HU, abdominal fat parameters, paraperirenal fat area, perirenal infiltration and severity of hydronephrosis with pre-treatment Non- Contrast Computed Tomography (NCCT) and stone density with radiography were evaluated by a radiologist. Four weeks after the last SWL; all patients were evaluated by plain X-ray and categorized as Stone Free (SF) and Residual Fragment (RF) group. 51 patients with renal stones and 88 patients with proximal ureteral stones were included in the study. 24 (47%) and 63 (71%) patients were classified as SFfor renal and ureteral stones respectively. Only stone size was an independent predictor for stone-free rates after SWL for renal and proximal ureteral stones on multivariate analysis. The Receiver Operating Characteristic (ROC) curves for renal calculi revealed that creatinine level, stone size, stone attitude, Houns-Field Unit (HU) and Skin-to-Stone Distance (SSD) were the predictive factors for the SWL outcome (p< 0.05). The ROC curve for ureteral calculi demonstrated that HU, stone size and attitude were the predictive factors (p< 0.05). All abdominal fat parameters and the stone density were not related to SWL failure. A large follow-up with more patients is essential to confirm the role of radiological parameters on the outcome of SWL.


The Older Adult Positivity Effect in Evaluations of Trustworthiness: Emotion Regulation or Cognitive Capacity?
An older adult positivity effect, i.e., the tendency for older adults to favor positive over negative stimulus information more than do younger adults, has been previously shown in attention, memory, and evaluations. This effect has been attributed to greater emotion regulation in older adults. In the case of attention and memory, this explanation has been supported by some evidence that the older adult positivity effect is most pronounced for negative stimuli, which would motivate emotion regulation, and that it is reduced by cognitive load, which would impede emotion regulation. We investigated whether greater older adult positivity in the case of evaluative responses to faces is also enhanced for negative stimuli and attenuated by cognitive load, as an emotion regulation explanation would predict. In two studies, younger and older adults rated trustworthiness of faces that varied in valence both under low and high cognitive load, with the latter manipulated by a distracting backwards counting task. In Study 1, face valence was manipulated by attractiveness (low /disfigured faces, medium, high/fashion models' faces). In Study 2, face valence was manipulated by trustworthiness (low, medium, high). Both studies revealed a significant older adult positivity effect. However, contrary to an emotion regulation account, this effect was not stronger for more negative faces, and cognitive load increased rather than decreased the rated trustworthiness of negatively valenced faces. Although inconsistent with emotion regulation, the latter effect is consistent with theory and research arguing that more cognitive resources are required to process negative stimuli, because they are more cognitively elaborated than positive ones. The finding that increased age and increased cognitive load both enhanced the positivity of trustworthy ratings suggests that the older adult positivity effect in evaluative ratings of faces may reflect age-related declines in cognitive capacity rather than increases in the regulation of negative emotions.


The evolution of reaction-diffusion controllers for minimally cognitive agents.
This article describes work carried out to investigate whether a classic reaction-diffusion (RD) system could be used to control a minimally cognitive animat. The RD system chosen was that first described by Gray and Scott, and the minimally cognitive behaviors were those used by Beer et al. involving the fixation and discrimination of diamond and circle shapes by a whiskered animat. A further task was added, which required the RD controllers to maintain and use a chemical memory. The parameters of these controllers were evolved using an evolutionary, or genetic, algorithm.


First Report of Zonate Leaf Spot of Glycine max Caused by Cristulariella moricola in Korea.
Soybean (Glycine max (L.) Merr.) is native to East Asia including Korea and is widely grown and consumed as an edible seed. In August 2011, following a prolonged period of cool and moist weather, zonate leaf spots were observed in local soybean (cultivar unknown) planted in a mountainous area of Goseong, central Korea. A voucher specimen was collected and entered at the Korea University herbarium (KUS-F26049). Initial symptoms included grayish green-to-grayish brown spots without border lines. As the lesions enlarged, they coalesced, leading to leaf blight and premature defoliation. Sporophores on the leaf lesions were dominantly hypophyllous, rarely epiphyllous, solitary, erect, easily detachable, and as long as 750 μm. The upper portion of the sporophores consisted of a pyramidal head that was ventricose, 275 to 500 μm long, and 80 to 160 μm wide. The fungus was isolated from leaf lesions and maintained on potato dextrose agar (PDA). Sclerotia were produced on PDA after 4 to 5 weeks at 18°C without light, but conidia were not observed in culture. The morphological and cultural characteristics were consistent with those of Cristulariella moricola (Hino) Redhead (2,3). An isolate was preserved in the Korean Agricultural Culture Collection (KACC46401). Genomic DNA was extracted with the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 453 bp was deposited in GenBank (Accession No. JQ036182). A BLAST search in GenBank revealed that the sequence showed an exact match with that of C. moricola from Acer negundo (JQ036181) and >99% similarity with that of Grovesinia pyramidalis, teleomorph of C. moricola from Juglans sp. (Z81433). To determine the pathogenicity of the fungus, sporophores with the pyramidal head were carefully detached from a lesion on the naturally infected leaflet with fine needles. Each sporophore was transferred individually onto four places of six detached healthy soybean leaflets. The leaflets were placed in humid chambers at 100% relative humidity and incubated at 16 to 20°C (4). Symptoms were observed after 2 days on all inoculated leaflets (one to four lesions/leaflet). The lesions enlarged rapidly and reached ~20 mm diameter in a week. A number of sporulating structures and immature sclerotia were formed on the abaxial surface of the leaf 2 weeks after inoculation. The pathogen was reisolated from lesions on the inoculated leaflets, confirming Koch's postulates. No symptoms were observed on the control leaflets kept in humid chambers for 2 weeks. C. moricola was known to cause zonate leaf spots and defoliation on a wide range of woody and annual plants (1), but not on G. max. To our knowledge, this is the first report of Cristulariella infection in cultivated soybeans. Since the infections may be limited to the mountainous area with low night temperature and high humidity, economic losses seem to be negligible. However, the disease could be a potential threat to the safe production of soybeans in areas with prolonged periods of cool and moist weather. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.arsgrin.gov/fungaldatabases/ , January 7, 2012. (2) H. B. Lee and C. J. Kim. Plant Dis. 86:440, 2002. (3) S. A. Redhead. Can. J. Bot. 53:700, 1975. (4) H. J. Su and S. C. Leu. Plant Dis. 67:915, 1983.


Gene exchange between two divergent species of the fungal human pathogen, Coccidioides.
The fungal genus Coccidioides is composed of two species, Coccidioides immitis and Coccidioides posadasii. These two species are the causal agents of coccidioidomycosis, a pulmonary disease also known as valley fever. The two species are thought to have shared genetic material due to gene exchange in spite of their long divergence. To quantify the magnitude of shared ancestry between them, we analyzed the genomes of a population sample from each species. Next, we inferred what is the expected size of shared haplotypes that might be inherited from the last common ancestor of the two species and find a cutoff to find what haplotypes have conclusively been exchanged between species. Finally, we precisely identified the breakpoints of the haplotypes that have crossed the species boundary and measure the allele frequency of each introgression in this sample. We find that introgressions are not uniformly distributed across the genome. Most, but not all, of the introgressions segregate at low frequency. Our results show that divergent species can share alleles, that species boundaries can be porous, and highlight the need for a systematic exploration of gene exchange in fungal species.


Camel No. 9 cigarette-marketing campaign targeted young teenage girls.
The 1998 Master Settlement Agreement (MSA) restricted tobacco industry advertising practices that targeted teens. To assess whether cigarette-advertising campaigns conducted after the MSA continue to influence smoking among adolescents. Participants were a national longitudinal cohort of 1036 adolescents (baseline age: 10-13 years) enrolled in a parenting study. Between 2003 and 2008, 5 sequential telephone interviews were conducted, including the participant's report of brand of "favorite" cigarette advertisement. The fifth interview was conducted after the start of RJ Reynolds' innovative "Camel No. 9" advertising campaign in 2007. Smoking outcome reported from the fifth survey. The response rate through the fifth survey was 71.8%. Teenagers who reported any favorite cigarette ad at baseline (mean age: 11.7 years) were 50% more likely to have smoked by the fifth interview (adjusted odds ratio: 1.5 [95% confidence interval: 1.0-2.3]). For boys, the proportion with a favorite ad was stable across all 5 surveys, as it was for girls across the first 4 surveys. However, after the start of the Camel No. 9 advertising campaign, the proportion of girls who reported a favorite ad increased by 10 percentage points, to 44%. The Camel brand accounted almost entirely for this increase, and the proportion of each gender that nominated the Marlboro brand remained relatively stable. After the MSA, adolescents continued to be responsive to cigarette advertising, and those who were responsive were more likely to start smoking. Recent RJ Reynolds advertising may be effectively targeting adolescent girls.


Entrainment of Breast Cell Lines Results in Rhythmic Fluctuations of MicroRNAs.
Circadian rhythms are essential for temporal (~24 h) regulation of molecular processes in diverse species. Dysregulation of circadian gene expression has been implicated in the pathogenesis of various disorders, including hypertension, diabetes, depression, and cancer. Recently, microRNAs (miRNAs) have been identified as critical modulators of gene expression post-transcriptionally, and perhaps involved in circadian clock architecture or their output functions. The aim of the present study is to explore the temporal expression of miRNAs among entrained breast cell lines. For this purpose, we evaluated the temporal (28 h) expression of 2006 miRNAs in MCF-10A, MCF-7, and MDA-MB-231 cells using microarrays after serum shock entrainment. We noted hundreds of miRNAs that exhibit rhythmic fluctuations in each breast cell line, and some of them across two or three cell lines. Afterwards, we validated the rhythmic profiles exhibited by miR-141-5p, miR-1225-5p, miR-17-5p, miR-222-5p, miR-769-3p, and miR-548ay-3p in the above cell lines, as well as in ZR-7530 and HCC-1954 using RT-qPCR. Our results show that serum shock entrainment in breast cells lines induces rhythmic fluctuations of distinct sets of miRNAs, which have the potential to be related to endogenous circadian clock, but extensive investigation is required to elucidate that connection.


A fundamental limitation of linearized algorithms for diffuse optical tomography.
Diffuse Optical Tomography is rapidly developing as a new imaging modality for characterizing the spatially varying optical properties of media which strongly scatter light (e.g. tissue). Numerous imaging algorithms exist, and more are being developed. Many of these algorithms rely on assumptions which linearize the relationship between the optical contrast and the perturbed signal. We show that this linear approximation makes quantitative imaging of spatially varying optical properties impossible. The explanation for this result is presented and the implication for Diffuse Optical Tomography is discussed.


Eating Decisions Based on Alertness Levels After a Single Night of Sleep Manipulation: A Randomized Clinical Trial.
To determine the relationship between an ecologically-relevant change in sleep behavior and its subsequent effects on daytime alertness and feeding behavior. Fifty healthy, young participants (10 male, 40 female) completed two 3-hour study sessions that were at least five days apart. The first session was a baseline evaluation. On the night prior to Session 2, the amount of time in bed was manipulated to be 60%-130% of the individual's habitual sleep time. Within both sessions, subjective (Stanford Sleepiness Scale) and objective (Psychomotor Vigilance Test) alertness were measured. During the middle of each session, a 40-minute ad libitum meal opportunity allowed participants to eat from eight different food items. Food healthfulness, caloric density, distribution, and number of calories were measured and compared to alertness levels. The induced variation in time in bed resulted in induced variation in both subjective and objective (p < .05) measures of alertness. Decreased subjective alertness was associated with increased total caloric consumption (p < .05), and a greater number of calories consumed from less healthy food (p < .05), as rated by both the investigators and by the participant. Decreased objective alertness was associated with less healthy food choices (p < .05), and the consumption of more food from the calorically-dense items (p < .05). Ecologically-relevant impairments in subjective and objective alertness are associated with increased caloric intake and dysfunctional eating decisions. People experiencing reduced alertness after modest sleep loss may be more willing to eat food they recognize as less healthful, and appear to prefer more calorically-dense foods.


Probing for hemispheric specialization for motor skill learning: a transcranial direct current stimulation study.
Convergent findings point to a left-sided specialization for the representation of learned actions in right-handed humans, but it is unknown whether analogous hemispheric specialization exists for motor skill learning. In the present study, we explored this question by comparing the effects of anodal transcranial direct current stimulation (tDCS) over either left or right motor cortex (M1) on motor skill learning in either hand, using a tDCS montage to better isolate stimulation to one hemisphere. Results were compared with those previously found with a montage more commonly used in the field. Six groups trained for three sessions on a visually guided sequential pinch force modulation task with their right or left hand and received right M1, left M1, or sham tDCS. A linear mixed-model analysis for motor skill showed a significant main effect for stimulation group (left M1, right M1, sham) but not for hand (right, left) or their interaction. Left M1 tDCS induced significantly greater skill learning than sham when hand data were combined, a result consistent not only with the hypothesized left hemisphere specialization for motor skill learning but also with possible increased left M1 responsiveness to tDCS. The unihemispheric montage effect size was one-half that of the more common montage, and subsequent power analysis indicated that 75 subjects per group would be needed to detect differences seen with only 12 subjects with the customary bihemispheric montage.


Polyphenols as potential indicators for drought tolerance in tea (Camellia sinensis L.).
Plant polyphenols have gained prominence in quality of plant products and in human health. An experiment was conducted to determine the association of tea polyphenols with water stress and their suitability as indicators for drought tolerance. The experiment was conducted in a 'rain-out' shelter, and consisted of six tea clones (BBK 35, TRFK 6/8, TRFK 76/1, TRFK 395/2, TRFK 31/30, and TRFK 311/287) and four levels of soil water contents (38, 30, 22, and 14% v/v), which were maintained for a period of 12 weeks. The treatments were arranged in a completely randomized design and replicated three times. Plant growth was monitored over 6 weeks, and a water stress index was calculated to determine water-stress tolerant clones. Total polyphenols in tea shoots was analyzed and a regression analysis done. The results indicate that declining soil water content (SWC) reduced both growth and content of polyphenols in tea. Tolerant clones maintained a high polyphenol content at low SWC, and also showed less fluctuation in phenolics when subjected to changes in SWC. There was significant (P<0.001) correlation of total polyphenol content with shoot growth and WSI of tea, and a linear relationship (r2=0.97) between SWC for tea and both, water stress index and shoot polyphenol content. We report that there is a potential to use polyphenols as indicators for selection of drought-tolerant tea cultivars.


The immunological synapse balances T cell receptor signaling and degradation.
The immunological synapse is a specialized cell-cell junction between T cell and antigen-presenting cell surfaces. It is characterized by a central cluster of antigen receptors, a ring of integrin family adhesion molecules, and temporal stability over hours. The role of this specific organization in signaling for T cell activation has been controversial. We use in vitro and in silico experiments to determine that the immunological synapse acts as a type of adaptive controller that both boosts T cell receptor triggering and attenuates strong signals.


Effects of trauma, economic hardship, and stress on neurocognition and everyday function in HIV.
The causes of neurocognitive and everyday functioning impairment among aging people living with HIV (PLWH) are multifactorial. Exposure to stress and trauma can result in neurocognitive deficits via activation of neurological and other biological mechanisms. PLWH (n = 122) and persons without HIV (n = 95), 35-65 years of age, completed four questionnaires that were used to generate a trauma, economic hardship (food insecurity and low socioeconomic status), and stress composite variable (TES). Participants also completed a comprehensive neuropsychological battery and standardized self-reports of activities of daily living (ADLs). We examined the independent and interactive effects of TES and HIV status on neurocognitive performance and ADL declines. PLWH had more traumatic events, more food insecurity, lower socioeconomic status, and higher perceived stress compared with HIV- individuals (all ps < .0001). Among PLWH, a higher composite TES score was associated with worse executive functioning (p = .02), worse learning (p = .02), worse working memory (p = .02), and more ADL declines (p < .0001), even after controlling for relevant demographic, psychiatric, substance use, and HIV disease covariates. On their own, individual TES components did not predict these outcomes. Conversely, no significant relationships were observed between TES and cognitive domains nor ADL declines among HIV- individuals. A composite score of trauma, economic hardship, and stress was significantly associated with worse neurocognitive performance and functional declines among PLWH. These adverse experiences may contribute to neurocognitive and daily functioning difficulties commonly observed among PLWH. Longitudinal studies are needed to elucidate the relationships between economic/psychosocial adversities and cognitive/functional outcomes over time, and examine potential mediators, such as inflammatory biomarkers. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Mental health services funding and development in KwaZulu-Natal: a tale of inequity and neglect.
Globally, a significant 'mental health gap' exists between the major burden of mental and substance use disorders and the provision of psychiatric and mental health services. As a signatory to the UN Convention on the Rights of Persons with Disabilities, South Africa has committed itself to transformation aimed at ending the inequities that characterise mental health service provision and 'closing the gap'. Budget allocations over a 5-year period to 6 psychiatric and 7 general hospitals in KwaZulu-Natal (KZN) are compared and current numbers of psychiatric beds and psychiatric personnel in that province are contrasted with the numbers required to comply with national norms. The mean increase in budget allocations to public psychiatric hospitals was 3.8% per annum, while that to general hospitals over the same period was 10.2% per annum. The median cumulative budget increase for psychiatric hospitals was significantly lower than that of general hospitals (Mann-Whitney U-test, p=0.001). No psychiatric hospitals received specific funding for tertiary services development. KZN has 25% of the acute psychiatric beds and 25% of the psychiatrists required to comply with national norms, with the most serious shortages experienced in northern KZN. There are 0.38 psychiatrists per 100 000 population in KZN. Inequitable funding, inadequate facilities and significant shortages of mental health professionals pervade the mental health and psychiatric services in KZN. There is little evidence of government abiding by its public commitments to redress the inequities that characterise mental health services.


Do Social Cognitive Theory constructs explain response heterogeneity with a physical activity behavioral intervention in multiple sclerosis?
Behavioral interventions based on Social Cognitive Theory (SCT) are efficacious for increasing both objectively-measured and self-reported physical activity in people with multiple sclerosis (MS). This study involved a secondary analysis of data focused on SCT constructs as correlates of individual-level changes (i.e., response heterogeneity) following a behavioral intervention. Twenty-two persons with MS who completed a 6-month SCT-based behavioral intervention for increasing physical activity were included in analyses. The intervention consisted of two primary components, namely a dedicated Internet website and one-on-one video chats with a behavioral coach. Outcomes included objectively-measured moderate-vigorous physical activity (MVPA) using Actigraph model GT3X+ accelerometers and self-reported physical activity using the Godin Leisure-Time Exercise Questionnaire (GLTEQ) as well as SCT variables of exercise self-efficacy, barriers self-efficacy, outcome expectations, goal setting and planning and facilitators/impediments. There was individual variability in physical activity change following the intervention. For example, 4/22 participants demonstrated a reduction in MVPA, 1/22 participants had no change, 9/22 participants had less than 0.5 standard deviation (SD) increase in MVPA, and 8/22 participants had an increase in MVPA of more than 0.5 SD. Baseline SCT variables, particularly outcome expectations, goal setting, planning, and barriers self-efficacy, correlated with increased physical activity. This study indicates that SCT variables correlate with the response heterogeneity associated with physical activity behavioral interventions, and this might inform the delivery of interventions consistent with Bandura's stepwise implementation model for optimizing the "fit" of an intervention based on SCT for maximizing treatment efficacy in MS.


Lamprey spinal interneurons and their roles in swimming activity.
An isolated lamprey spinal cord generates rhythmic ventral root bursting that closely resembles swimming activity: ventral roots on opposite sides of the spinal cord burst in alternation, and rostral ventral roots lead caudal ventral roots. This rhythmic activity is induced by superfusion of the spinal cord with an excitatory amino acid and is called 'fictive' swimming. Three main types of spinal interneurons that are active during fictive swimming have been characterized: small excitatory interneurons with ipsilateral axons, large inhibitory interneurons with ipsilateral descending axons (lateral interneurons), and inhibitory commissural interneurons. The synaptic connectivities of these inter-neurons can account for the pattern of motoneuron excitation and inhibition occurring during fictive swimming, and it has been proposed that the synaptic interactions of these spinal neurons form the unit segmental oscillators of the swim network. Computer modeling has confirmed that this network can generate rhythmic activity resembling fictive swimming. The core of the model is the reciprocal inhibition between commissural interneurons on opposite sides of the cord. Evidence that the commissural interneurons are essential to rhythm generation comes from lesion studies in which the spinal cord was split down the midline and also from photo-ablation studies in which commissural interneurons were inactivated by illumination after retrograde labeling with a photo-toxic tracer. In both types of experiments, rhythmic activity of fictive swimming can be abolished, supporting the view that the commissural interneurons are necessary for rhythmogenesis.


Use of model soil colloids to evaluate adsorption of phenanthrene and its mobilization by different solutions.
Inorganic (a montmorillonite; a complex of montmorillonite and aluminium hydroxide) and organo-mineral model soil colloids (a complex of montmorillonite, aluminium hydroxide and humic acid) were tested for their capacity to adsorb and desorb phenanthrene. The adsorption-desorption isotherms demonstrated that the inorganic surfaces were able to bind the pollutant, although to a lesser extent than the organo-mineral complex. The capacity of different aqueous solutions to desorb phenanthrene after 1, 7, and 30 days' contact with the model colloids was tested in order to assess their efficacy with respect to the aging effect. The solutions used were 0.01 N CaCl(2), dissolved soil organic matter (DOM), a model root exudates solution (aqueous solution of saccharose, fructose, glucose, fumaric acid, ferrulic acid, and leucine), and three commercial surfactants (sodium dodecyl sulfate, Tween 20, and didecyldimethylammonium bromide). The desorptive capacity of these solutions were compared with that of organic solutions (water-ethanol and n-hexane-ethanol). More than 85% phenanthrene was desorbed after 30 days aging by organic solutions. Among the aqueous solutions, the best results were obtained with dissolved soil organic matter solution and the model root exudates solution which were able to desorb up to 50% phenanthrene after 30 days aging. Such naturally occurring aqueous solutions should be preferred to synthetic compounds to mobilize phenanthrene in contaminated sites without further contaminating the environment.


Cranial neural crest cells form corridors prefiguring sensory neuroblast migration.
The majority of cranial sensory neurons originate in placodes in the surface ectoderm, migrating to form ganglia that connect to the central nervous system (CNS). Interactions between inward-migrating sensory neuroblasts and emigrant cranial neural crest cells (NCCs) play a role in coordinating this process, but how the relationship between these two cell populations is established is not clear. Here, we demonstrate that NCCs generate corridors delineating the path of migratory neuroblasts between the placode and CNS in both chick and mouse. In vitro analysis shows that NCCs are not essential for neuroblast migration, yet act as a superior substrate to mesoderm, suggesting provision of a corridor through a less-permissive mesodermal territory. Early organisation of NCC corridors occurs prior to sensory neurogenesis and can be recapitulated in vitro; however, NCC extension to the placode requires placodal neurogenesis, demonstrating reciprocal interactions. Together, our data indicate that NCC corridors impose physical organisation for precise ganglion formation and connection to the CNS, providing a local environment to enclose migrating neuroblasts and axonal processes as they migrate through a non-neural territory.


Mitosis inhibits DNA double-strand break repair to guard against telomere fusions.
Mitotic cells inactivate DNA double-strand break (DSB) repair, but the rationale behind this suppression remains unknown. Here, we unravel how mitosis blocks DSB repair and determine the consequences of repair reactivation. Mitotic kinases phosphorylate the E3 ubiquitin ligase RNF8 and the nonhomologous end joining factor 53BP1 to inhibit their recruitment to DSB-flanking chromatin. Restoration of RNF8 and 53BP1 accumulation at mitotic DSB sites activates DNA repair but is, paradoxically, deleterious. Aberrantly controlled mitotic DSB repair leads to Aurora B kinase-dependent sister telomere fusions that produce dicentric chromosomes and aneuploidy, especially in the presence of exogenous genotoxic stress. We conclude that the capacity of mitotic DSB repair to destabilize the genome explains the necessity for its suppression during mitosis, principally due to the fusogenic potential of mitotic telomeres.


Expanding medicinal chemistry into 3D space: metallofragments as 3D scaffolds for fragment-based drug discovery.
Fragment-based drug discovery (FBDD) is a powerful strategy for the identification of new bioactive molecules. FBDD relies on fragment libraries, generally of modest size, but of high chemical diversity. Although good chemical diversity in FBDD libraries has been achieved in many respects, achieving shape diversity - particularly fragments with three-dimensional (3D) structures - has remained challenging. A recent analysis revealed that >75% of all conventional, organic fragments are predominantly 1D or 2D in shape. However, 3D fragments are desired because molecular shape is one of the most important factors in molecular recognition by a biomolecule. To address this challenge, the use of inert metal complexes, so-called 'metallofragments' (mFs), to construct a 3D fragment library is introduced. A modest library of 71 compounds has been prepared with rich shape diversity as gauged by normalized principle moment of inertia (PMI) analysis. PMI analysis shows that these metallofragments occupy an area of fragment space that is unique and highly underrepresented when compared to conventional organic fragment libraries that are comprised of orders of magnitude more molecules. The potential value of this metallofragment library is demonstrated by screening against several different types of proteins, including an antiviral, an antibacterial, and an anticancer target. The suitability of the metallofragments for future hit-to-lead development was validated through the determination of IC50 and thermal shift values for select fragments against several proteins. These findings demonstrate the utility of metallofragment libraries as a means of accessing underutilized 3D fragment space for FBDD against a variety of protein targets.


Genotypically defined β-lactamase-negative ampicillin-resistant isolates of non-typable Haemophilus influenzae are associated with increased invasion of bronchial epithelial cells in vitro.
The aim of the study was to investigate the association between the presence of altered penicillin-binding protein 3 (PBP3) in non-typable Haemophilus influenzae (NTHi) and an increased capacity to invade bronchial epithelial cells in vitro. A collection of 40 clinical isolates of NTHi comprised of 20 with normal PBP3 and 20 with altered PBP3 (defined by an N526K substitution) was established. The isolates were tested for the ability to invade bronchial epithelial cells in vitro using a 4 h gentamicin survival assay. Invasion was measured as the percentage of intracellular organisms relative to the initial inoculum. The mean invasion rate was 0.00-14.79 % in the normal PBP3 isolates and 0.02-36.69 % in the altered PBP3 isolates. The altered PBP3 isolates had a higher (P = 0.003) mean invasion rate (6.86 %, n = 20) than the normal PBP3 isolates (1.31 %, n = 20). Subsequently, two variants of altered PBP3 (transformant 1, N526K; transformant 2, M377I, S385T, L389F and N526K) were cloned into three of the initial isolates (parents) with normal PBP3 and relatively low invasive ability, and the parents and transformants tested for invasion as above. There was no difference (P = 0.89) in the mean invasion rates for the parents (0.81 %, n = 3), transformants 1 (0.90 %, n = 3) and transformants 2 (1.38 %, n = 3). There was an association between the presence of altered PBP3 in NTHi and an increased capacity to invade BEAS-2B cells in vitro, but cloning experiments suggested that the altered PBP3 was not involved directly in enhanced invasion.


Affective and cognitive reactions to subliminal flicker from fluorescent lighting.
This study renews the classical concept of subliminal perception (Peirce & Jastrow, 1884) by investigating the impact of subliminal flicker from fluorescent lighting on affect and cognitive performance. It was predicted that low compared to high frequency lighting (latter compared to former emits non-flickering light) would evoke larger changes in affective states and also impair cognitive performance. Subjects reported high rather than low frequency lighting to be more pleasant, which, in turn, enhanced their problem solving performance. This suggests that sensory processing can take place outside of conscious awareness resulting in conscious emotional consequences; indicating a role of affect in subliminal/implicit perception, and that positive affect may facilitate cognitive task performance.


The E3 ubiquitin ligases Hrd1 and gp78 bind to and promote cholera toxin retro-translocation.
To cause disease, cholera toxin (CT) is transported from the cell surface to the endoplasmic reticulum (ER) lumen where the catalytic CTA1 subunit retro-translocates to the cytosol to induce pathological water secretion. Two retro-translocon components are the Derlins and ER-associated multi-spanning E3 ubiquitin ligases including Hrd1 and gp78. We demonstrated previously that Derlin-1 facilitates CTA1 retro-translocation. However, as CTA1 is neither ubiquitinated on lysines nor at its N-terminus, the role of E3 ligases in toxin retro-translocation is unclear. Here, we show that expression of mutant Hrd1 and gp78 and a mutant E2-conjugating enzyme dedicated to retro-translocation (Ube2g2) decrease CTA1 retro-translocation. Hrd1 knockdown also attenuated toxin retro-translocation. Binding studies demonstrate that Hrd1 and gp78 interact with CT and protein disulfide isomerase, an ER chaperone that unfolds CTA1 to initiate translocation. Moreover, we find that the toxin's association with Hrd1 and gp78 is blocked by dominant-negative Derlin-1, suggesting that CT is targeted initially to Derlin-1 and then transferred to Hrd1 and gp78. These data demonstrate a role of the E3 ubiquitin ligases in CTA1 retro-translocation, implicate a sequence of events experienced by the toxin on the ER membrane, and raise the possibility that ubiquitination is involved in the transport process.


Comparison of fusion rates following transforaminal lumbar interbody fusion using polyetheretherketone cages or titanium cages with transpedicular instrumentation.
Compared to titanium cage, polyetheretherketone (PEEK) cage with pedicle screw fixation has been increasingly used in transforaminal lumbar interbody fusion (TLIF). However, there is insufficient evidence supporting the superiority of PEEK cages over titanium cages as optimal TLIF spacers. The aim of this study was to compare the clinical and radiographic outcomes of patients at a 2-year follow-up after undergoing instrumented TLIF in which either a PEEK cage or a titanium cage was implanted. We retrospectively analyzed prospectively collected 48 patients who underwent single-level TLIF in which the first 23 patients received a titanium cage and the 25 patients received a PEEK cage. Patient demographics, clinical outcomes, and radiographic imaging were studied. Improvement of clinical outcomes was comparable between the two groups. Based on the criteria using computed tomography, 96 % in the Titanium group and 64 % in the PEEK group showed fusion at 12 months. At 24 months, fusion rate in the Titanium group was increased to 100 %, while PEEK group showed 76 % of fusion rate. In the PEEK group, vertebral osteolysis was noted in 60 % of the cases with nonunion. This abnormal finding was not observed in the Titanium group. Vertebral osteolysis was significantly associated with nonunion. The superiority of PEEK cages over titanium cages for bony fusion was not demonstrated. Additionally, we found unfavorable radiographic findings in the cases with a PEEK cage, which may lead to nonunion. Improvement in biocompatibility of a PEEK cage will be needed to increase the fusion rate.


Diet and dental topography in pitheciine seed predators.
Pitheciines (Pithecia, Chiropotes, and Cacajao) are a specialized clade of Neotropical seed predators that exhibit postcanine teeth with low and rounded cusps and highly crenulated occlusal surface enamel. Data on feeding ecology show that Pithecia consumes proportionally more leaves than other pitheciine species, and comparative studies demonstrate its greater molar relief and relative shearing potential. However, data on pitheciine food mechanics show that Pithecia masticates seeds with greater crushing resistance than those preferred by Chiropotes. This variation predicts an opposing morphology characterized by low and more rounded occlusal surfaces in Pithecia. We build on previous research using new methods for molar surface shape quantification by examining pitheciine second molar shearing crest length, occlusal relief, surface complexity, and surface curvature relative to nonseed specializing platyrrhines and within the context of the observed interspecific variation in pitheciine feeding ecology. Consistent with the previous analyses, our findings demonstrate that pitheciine molars exhibit low shearing, relief, and curvature compared with nonseed predators, independent of phylogeny. Pitheciines also exhibit highly "complex" occlusal topography that promotes the efficient breakdown of tough seed tissues. Overall, Pithecia, Chiropotes, and Cacajao share a similar topographic pattern, suggesting adaptation to foods with similar structural and/or mechanical properties. However, Cacajao differs in surface complexity, which reflects some variation in its feeding ecology. Contrary to the predictions, Pithecia and Chiropotes do not differ in any of the topographic variables examined. The range of demands imposed on the postcanine teeth of Pithecia might therefore select for an average topography, one that converges on that of Chiropotes.


Diarrhoea in a large prospective cohort of European travellers to resource-limited destinations.
Incidence rates of travellers' diarrhoea (TD) need to be updated and risk factors are insufficiently known. Between July 2006 and January 2008 adult customers of our Centre for Travel Health travelling to a resource-limited country for the duration of 1 to 8 weeks were invited to participate in a prospective cohort study. They received one questionnaire pre-travel and a second one immediately post-travel. First two-week incidence rates were calculated for TD episodes and a risk assessment was made including demographic and travel-related variables, medical history and behavioural factors. Among the 3100 persons recruited, 2800 could be investigated, resulting in a participation rate of 89.2%. The first two-weeks incidence for classic TD was 26.2% (95%CI 24.5-27.8). The highest rates were found for Central Africa (29.6%, 95% CI 12.4-46.8), the Indian subcontinent (26.3%, 95%CI 2.3-30.2) and West Africa (21.5%, 95%CI 14.9-28.1). Median TD duration was 2 days (range 1-90). The majority treated TD with loperamide (57.6%), while a small proportion used probiotics (23.0%) and antibiotics (6.8%). Multiple logistic regression analysis on any TD to determine risk factors showed that a resolved diarrhoeal episode experienced in the 4 months pre-travel (OR 2.03, 95%CI 1.59-2.54), antidepressive comedication (OR 2.11, 95%CI 1.17-3.80), allergic asthma (OR 1.67, 95%CI 1.10-2.54), and reporting TD-independent fever (OR 6.56, 95%CI 3.06-14.04) were the most prominent risk factors of TD. TD remains a frequent travel disease, but there is a decreasing trend in the incidence rate. Patients with a history of allergic asthma, pre-travel diarrhoea, or of TD-independent fever were more likely to develop TD while abroad.


Long non-coding RNA MALAT-1 overexpression predicts tumor recurrence of hepatocellular carcinoma after liver transplantation.
Metastasis-associated lung adenocarcinoma transcript 1(MALAT1), a long non-coding RNA (lncRNA), is up-regulated in many solid tumors and associated with cancer metastasis and recurrence. However, its role in hepatocellular carcinoma (HCC) remains poorly understood. In the present study, we evaluated the expression of MALAT1 by quantitative real-time PCR in 9 liver cancer cell lines and 112 HCC cases including 60 cases who received liver transplantation (LT) with complete follow-up data. Moreover, small interfering RNA (siRNA) was used to inhibit MALAT1 expression to investigate its biological role in tumor progression. We found that MALAT1 was up-regulated in both cell lines and clinical tissue samples. Patients with high expression level of MALAT1 had a significantly increased risk of tumor recurrence after LT, particularly in patients who exceeded the Milan criteria. On multivariate analysis, MALAT1 was an independent prognostic factor for predicting HCC recurrence (hazard ratio, 3.280, P = 0.003).In addition, inhibition of MALAT1 in HepG2 cells could effectively reduce cell viability, motility, invasiveness, and increase the sensitivity to apoptosis. Our data suggest that lncRNA MALAT1 play an important role in tumor progression and could be a novel biomarker for predicting tumor recurrence after LT and serve as a promising therapeutic target.


The HARMONIC trial: study protocol for a randomised controlled feasibility trial of Shaping Healthy Minds-a modular transdiagnostic intervention for mood, stressor-related and anxiety disorders in adults.
Anxiety, mood and trauma-related disorders are common, affecting up to 20% of adults. Many of these individuals will experience symptoms of more than one disorder as diagnostically defined. However, most psychological treatments focus on individual disorders and are less effective for those who experience comorbid disorders. The Healthy and Resilient Mind Programme: Building Blocks for Mental Wellbeing (HARMONIC) trial introduces a novel transdiagnostic intervention (Shaping Healthy Minds (SHM)), which synthesises several evidence-based treatment techniques to address the gap in effective interventions for people with complex and comorbid difficulties. This early phase trial aims to estimate the efficacy and feasibility of the transdiagnostic intervention in preparation for a later-phase randomised controlled trial, and to explore mechanisms of change. We outline a patient-level two-arm randomised controlled trial (HARMONIC) that compares SHM to treatment-as-usual for individuals aged >18 years (n=50) with comorbid mood, anxiety, obsessive-compulsive or trauma/stressor disorders diagnoses, recruited from outpatient psychological services within the UK National Health Service (NHS). The co-primary outcomes will be 3-month follow-up scores on self-report measures of depressive symptoms, anxiety symptoms, and disability and functional impairment. Secondary outcomes include changes in symptoms linked to individual disorders. We will assess the feasibility and acceptability of SHM, the utility of proposed outcome measures, and refine the treatment manuals in preparation for a later-phase trial. This trial protocol has been approved by the Health Research Authority of the NHS of the UK (East of England, Reference: 16/EE/0095). We anticipate that trial findings will inform future revisions of clinical guidelines for numerous forms of mood, anxiety and stressor-related disorders. Findings will be disseminated broadly via peer-reviewed empirical journal articles, conference presentations, clinical workshops and a trial website. NCT03143634; Pre-results.


The fibrillin microfibril scaffold: A niche for growth factors and mechanosensation?
The fibrillins, large extracellular matrix molecules, are polymerized to form "microfibrils." The fibrillin microfibril scaffold is populated by microfibril-associated proteins and by growth factors, which are likely to be latent. The scaffold, associated proteins, and bound growth factors, together with cellular receptors that can sense the microfibril matrix, constitute the fibrillin microenvironment. Activation of TGFβ signaling is associated with the Marfan syndrome, which is caused by mutations in fibrillin-1. Today we know that mutations in fibrillin-1 cause the Marfan syndrome as well as Weill-Marchesani syndrome (and other acromelic dysplasias) and result in opposite clinical phenotypes: tall or short stature; arachnodactyly or brachydactyly; joint hypermobility or stiff joints; hypomuscularity or hypermuscularity. We also know that these different syndromes are associated with different structural abnormalities in the fibrillin microfibril scaffold and perhaps with specific cellular receptors (mechanosensors). How does the microenvironment, framed by the microfibril scaffold and populated by latent growth factors, work? We must await future investigations for the molecular and cellular mechanisms that will answer this question. However, today we can appreciate the importance of the fibrillin microfibril niche as a contextual environment for growth factor signaling and potentially for mechanosensation.


Isolation of Dickeya dadantii strains from potato disease and biocontrol by their bacteriophages.
One of the most economically important bacterial pathogens of plants and plant products is Dickeya dadantii. This bacterium causes soft rot disease in tubers and other parts of the potato and other plants of the Solanaceae family. The application of restricted host range bacteriophages as biocontrol agents has recently gained widespread interest. This study purposed to isolate the infectious agent of the potato and evaluate its biocontrol by bacteriophages. Two phytopathogenic strains were isolated from infected potatoes, identified based on biochemical and 16S rRNA gene sequencing, and submitted to GenBank as D. dadantii strain pis3 (accession no. HQ423668) and D. dadantii strain sip4 (accession no. HQ423669). Their bacteriophages were isolated from Caspian Sea water by enriching the water filtrate with D. dadantii strains as hosts using spot or overlay methods. On the basis of morphotypes, the isolated bacteriophages were identified as members of the Myoviridae and Siphoviridae families and could inhibit the growth of antibiotic resistant D. dadantii strains in culture medium. Moreover, in Dickeya infected plants treated with bacteriophage, no disease progression was detected. No significant difference was seen between phage-treated and control plants. Thus, isolated bacteriophages can be suggested for the biocontrol of plant disease caused by Dickeya strains.


Student evaluation of a Clinical Self-Study Laboratory.
Being clinically competent is an essential component of any nursing curricula to ensure that on completion the nursing graduates are able to practice safely and effectively as a nurse. Clinical Self-Study Laboratories have become increasingly popular in nursing education in recent years as a way of ensuring a variety of skills are taught to the nursing students. These clinical skills laboratories are used to teach communication and interpersonal skills, psychomotor skills, promote the development of collaborative skills required in nursing as well as being able to help integrate theory and practice [Morgan, R., 2006. Using clinical skills laboratories to promote theory-practice integration during first practice placement: an Irish perspective. Journal of Clinical Nursing 15, 155-161]. However, it is essential that these programs be subject to routine quality assurance and effectiveness evaluation. The purpose of the study was to conduct a process evaluation of the implementation of the Clinical Self-Study Laboratory (CSSL) in the School of Nursing, University of KwaZulu-Natal. The process evaluation included a descriptive quantitative satisfaction survey of first and third year Bachelor of Nursing students, a checklist to assess the quality of the CSSL equipment and a retrospective record review of utilisation of the CSSL by students. The CSSL appears to be well implemented and utilized by students. Students reported that it was accessible and that the equipment in the laboratory was of a reasonable quality. A few students reported dissatisfaction with some of the equipment and requested that more equipment and trained personnel be made available.


Dissociable neural mechanisms track evidence accumulation for selection of attention versus action.
Decision-making is typically studied as a sequential process from the selection of what to attend (e.g., between possible tasks, stimuli, or stimulus attributes) to which actions to take based on the attended information. However, people often process information across these various levels in parallel. Here we scan participants while they simultaneously weigh how much to attend to two dynamic stimulus attributes and what response to give. Regions of the prefrontal cortex track information about the stimulus attributes in dissociable ways, related to either the predicted reward (ventromedial prefrontal cortex) or the degree to which that attribute is being attended (dorsal anterior cingulate cortex, dACC). Within the dACC, adjacent regions track correlates of uncertainty at different levels of the decision, regarding what to attend versus how to respond. These findings bridge research on perceptual and value-based decision-making, demonstrating that people dynamically integrate information in parallel across different levels of decision-making.


Controlling supercurrents and their spatial distribution in ferromagnets.
Spin-triplet Cooper pairs induced in ferromagnets form the centrepiece of the emerging field of superconducting spintronics. Usually the focus is on the spin-polarization of the triplets, potentially enabling low-dissipation magnetization switching. However, the magnetic texture which provides the fundamental mechanism for generating triplets also permits control over the spatial distribution of supercurrent. Here we demonstrate the tailoring of distinct supercurrent pathways in the ferromagnetic barrier of a Josephson junction. We combine micromagnetic simulations with three-dimensional supercurrent calculations to design a disk-shaped structure with a ferromagnetic vortex which induces two transport channels across the junction. By using superconducting quantum interferometry, we show the existence of two channels. Moreover, we show how the supercurrent can be controlled by moving the vortex with a magnetic field. This approach paves the way for supercurrent paths to be dynamically reconfigured in order to switch between different functionalities in the same device.


Structure-function relationships in Anabaena ferredoxin: correlations between X-ray crystal structures, reduction potentials, and rate constants of electron transfer to ferredoxin:NADP+ reductase for site-specific ferredoxin mutants.
A combination of structural, thermodynamic, and transient kinetic data on wild-type and mutant Anabaena vegetative cell ferredoxins has been used to investigate the nature of the protein-protein interactions leading to electron transfer from reduced ferredoxin to oxidized ferredoxin:NADP+ reductase (FNR). We have determined the reduction potentials of wild-type vegetative ferredoxin, heterocyst ferredoxin, and 12 site-specific mutants at seven surface residues of vegetative ferredoxin, as well as the one- and two-electron reduction potentials of FNR, both alone and in complexes with wild-type and three mutant ferredoxins. X-ray crystallographic structure determinations have been carried out for six of the ferredoxin mutants. None of the mutants showed significant structural changes in the immediate vicinity of the [2Fe-2S] cluster, despite large decreases in electron-transfer reactivity (for E94K and S47A) and sizable increases in reduction potential (80 mV for E94K and 47 mV for S47A). Furthermore, the relatively small changes in Calpha backbone atom positions which were observed in these mutants do not correlate with the kinetic and thermodynamic properties. In sharp contrast to the S47A mutant, S47T retains electron-transfer activity, and its reduction potential is 100 mV more negative than that of the S47A mutant, implicating the importance of the hydrogen bond which exists between the side chain hydroxyl group of S47 and the side chain carboxyl oxygen of E94. Other ferredoxin mutations that alter both reduction potential and electron-transfer reactivity are E94Q, F65A, and F65I, whereas D62K, D68K, Q70K, E94D, and F65Y have reduction potentials and electron-transfer reactivity that are similar to those of wild-type ferredoxin. In electrostatic complexes with recombinant FNR, three of the kinetically impaired ferredoxin mutants, as did wild-type ferredoxin, induced large (approximately 40 mV) positive shifts in the reduction potential of the flavoprotein, thereby making electron transfer thermodynamically feasible. On the basis of these observations, we conclude that nonconservative mutations of three critical residues (S47, F65, and E94) on the surface of ferredoxin have large parallel effects on both the reduction potential and the electron-transfer reactivity of the [2Fe-2S] cluster and that the reduction potential changes are not the principal factor governing electron-transfer reactivity. Rather, the kinetic properties are most likely controlled by the specific orientations of the proteins within the transient electron-transfer complex.


Results of surgery for high-risk prostate cancer.
Surgery for high-risk prostate cancer (PCa) is applied frequently nowadays. Nevertheless, this approach is still surrounded by many controversies. The present review discusses the most recent literature regarding surgery for high-risk PCa. As there is no standard definition of high-risk PCa, outcome comparison between series and treatment approaches is hampered. Nevertheless, recent radical prostatectomy series have shown excellent cancer-specific survival in patients with high-risk PCa. Even for very-high-risk PCa (cT3b-T4 or any cT, N1), surgery may be applied to highly selected patients as a first step of a multimodality approach. Recent experience with robot-assisted surgery opens new possibilities for a minimally invasive approach in this field.Patient selection for surgery was also addressed in recent studies. Excellent cancer-specific survival is seen when specimen-confined PCa is found at final histopathology; a recently published nomogram enables the prediction of specimen-confined disease. Another issue in high-risk PCa is the impact of age and comorbidities on cancer-specific and overall mortality. In a recent study, it was shown that patients with low comorbidity scores, even when at least 70 years old, had a significant risk of dying from their cancer and may benefit most from a surgical approach. A modified extended pelvic lymphadenectomy template was presented, providing optimal removal of positive lymph nodes. Radical prostatectomy with extended pelvic lymphadenectomy delivers very good cancer-related outcomes in high-risk and very-high-risk PCa, often within a multimodal approach. Minimally invasive surgery and improved patient selection will be key to further improve oncological and functional outcomes.