Multi-protein complexes in the cis Golgi of Saccharomyces cerevisiae with alpha-1,6-mannosyltransferase activity.
Anp1p, Van1p and Mnn9p constitute a family of membrane proteins required for proper Golgi function in Saccharomyces cerevisiae. We demonstrate that these proteins colocalize within the cis Golgi, and that they are physically associated in two distinct complexes, both of which contain Mnn9p. Furthermore, we identify two new proteins in the Anp1p-Mnn9p-containing complex which have homology to known glycosyltransferases. Both protein complexes have alpha-1, 6-mannosyltransferase activity, forming a series of poly-mannose structures. These reaction products also contain some alpha-1, 2-linked mannose residues. Our data suggest that these two multi-protein complexes are responsible for the synthesis and initial branching of the long alpha-1,6-linked backbone of the hypermannose structure attached to many yeast glycoproteins.


Mercury Contamination in Riverine Sediments and Fish Associated with Artisanal and Small-Scale Gold Mining in Madre de Dios, Peru.
Artisanal and small-scale gold mining (ASGM) in Madre de Dios, Peru, continues to expand rapidly, raising concerns about increases in loading of mercury (Hg) to the environment. We measured physicochemical parameters in water and sampled and analyzed sediments and fish from multiple sites along one ASGM-impacted river and two unimpacted rivers in the region to examine whether Hg concentrations were elevated and possibly related to ASGM activity. We also analyzed the 308 fish samples, representing 36 species, for stable isotopes (δ15N and δ13C) to estimate their trophic position. Trophic position was positively correlated with the log-transformed Hg concentrations in fish among all sites. There was a lack of relationship between Hg concentrations in fish and either Hg concentrations in sediments or ASGM activity among sites, suggesting that fish Hg concentrations alone is not an ideal bioindicator of site-specific Hg contamination in the region. Fish Hg concentrations were not elevated in the ASGM-impacted river relative to the other two rivers; however, sediment Hg concentrations were highest in the ASGM-impacted river. Degraded habitat conditions and commensurate shifts in fish species and ecological processes may influence Hg bioaccumulation in the ASGM-impacted river. More research is needed on food web dynamics in the region to elucidate any effects caused by ASGM, especially through feeding relationships and food sources.


The atrazine metabolite diaminochlorotriazine suppresses LH release from murine LβT2 cells by suppressing GnRH-induced intracellular calcium transients.
The primary metabolite of the herbicide atrazine (ATRA), diaminochlorotriazine (DACT), has been suggested to cause disruption in the hypothalamic-pituitary-gonadal axis leading to inhibition of luteinizing hormone (LH) release. DACT is a reactive electrophile known to form covalent protein adducts both in vitro and in vivo following ATRA exposure and maybe targeting proteins involved in GnRH-induced calcium signaling and subsequent LH release. To test this hypothesis, LβT2 pituitary cells were exposed to 300 μM DACT for 24 hrs and examined by fluorescence microscopy for GnRH-induced changes in intracellular calcium and LH release. LβT2 cells exposed to DACT had markedly diminished GnRH-induced intracellular calcium transients and a significant decreased LH release in response to GnRH. DACT appeared to cause a selective decrease in caffeine-sensitive ryanodine receptor-operated calcium stores in LβT2 cells, rather than in thapsigargin-sensitive ER calcium stores. This sensitivity correlated with the formation of covalent protein adducts by DACT, as determined by mass spectrometry. ERp57 was identified by mass spectrometry as a target of DACT adduction in the ER that could potentially mediate the effects of DACT on inhibition of GnRH-induced calcium signaling and inhibition of LH release. Intracellular calcium responses to GnRH and release of LH were restored in DACT-treated cells with the addition of a calcium ionophore (A23187). These data suggest that DACT forms adducts on proteins involved in calcium handling within the ER and that dysfunction in this critical signaling system is associated with loss of normal sensitivity to GnRH and subsequent decreased release of LH.


Monitoring adipose tissue blood flow in man: a comparison between the (133)xenon washout method and microdialysis.
Adipose tissue blood flow (ATBF) increases after meal intake and a failure to regulate ATBF in the postprandial period seems to be a feature of insulin resistance and obesity. ATBF can be measured quantitatively by the (133)Xe washout technique, but the microdialysis ethanol escape method has also been employed to detect relative changes in ATBF. We compared (133)Xe washout and the recovery of exogenous ethanol and endogenous urea by microdialysis in abdominal subcutaneous adipose tissue, after physiological stimulation of ATBF by ingestion of oral glucose (75 g) in eight healthy people (age 23-52 y, body mass index (BMI) 19.4-29.6 kg/m(2)). The ATBF response was heterogeneous. In subjects responding vigorously to the stimulus as measured by (133)Xe washout, the microdialysis ethanol escape was increased (indicating an increase in ATBF). An increased recovery of urea was observed, also indicating an increase in ATBF. The recovery of both small molecules was delayed compared with increased blood flow and failed to return to baseline in response to a rapid decline in ATBF. We conclude that the (133)Xe washout technique is more responsive to physiological change in ATBF than ethanol escape or urea recovery by microdialysis.


Response of Millepora alcicornis (Milleporina: Milleporidae) to two bleaching events at Puerto Morelos reef, Mexican Caribbean.
Two naturally occurring colonies of Millepora alcicornis were monitored during 1997 and 1998, both years in which this species bleached in the Mexican Caribbean. One colony (HL) was naturally exposed to a high light environment and another nearby colony (LL) was exposed to 5.9 times lower light levels due to shadowing by a pier. For 10 days in August 1997, seawater temperatures in the surrounding reef lagoon rose up to 1.5 degrees C above the 6-year August average. The HL colony bleached to white during this period, whereas, the LL colony remained dark-brown colored. The HL colony recovered its normal dark-brown coloration (reversible bleaching) within several weeks, during which time the seawater temperatures returned to average. The following year, for 10 days, seawater temperatures rose up to 3 degrees C above the 7-year August average and both colonies bleached to white and neither colony recovered (irreversible bleaching). Both colonies were rapidly overgrown by algae and hydroids and, as of June 2003, no recovery has taken place. Prior to the 1997 bleaching, experiments using solar radiation showed that the quantum yield of photosystem II charge separation of branches from HL and LL colonies were affected for several hours by exposure to ultraviolet radiation (UVR, 280 to 400 nm), but recovered by the same evening, suggesting that UVR does not have long-term effects on photochemistry in M. alcicornis. In situ effective quantum yield of photosystem II charge separation (deltaF/Fm') measurements before the 1998 bleaching event indicate that both colonies were healthy in terms of the physiological status of their endosymbionts. During and after the 1998 bleaching event, both colonies showed a reduction in deltaF/Fm' and consequently an increase in excitation pressure on photosystem II. The data suggest that temperature is not the only factor that causes bleaching and that solar radiation may play an important role in coral bleaching.


The ETS family transcription factor PU.1 is necessary for the maintenance of fetal liver hematopoietic stem cells.
PU.1 is a member of the ETS family of transcription factors and is required for the development of multiple hematopoietic lineages. PU.1(-/-) mice die from hematopoietic failure at about embryonic day 18.5 (e18.5) and show a complete absence of B cells, mature T cells, and macrophages. This phenotype suggests that PU.1 may function at the level of the hematopoietic stem cell (HSC) or a multilineage progenitor. To investigate the role of PU.1 in the regulation of HSCs, PU.1(-/-) embryos were analyzed at various stages of embryonic development. The absolute number and frequency of HSCs were determined by flow cytometric analysis of c-Kit(+)Thy-1.1(lo)Lin(-)Sca-1(+) (KTLS) cells. We found that KTLS cells were absent or severely reduced in PU.1(-/-) fetal liver from e12.5 to e15.5. Progenitor cells with a c-Kit(+)Lin(-)AA4.1(+) and c-Kit(+)Lin(-)CD34(+) phenotype were also severely reduced. In addition, PU.1(-/-) fetal liver at e14.5 lacked common myeloid progenitors (CMPs) and granulocyte-macrophage progenitors (GMPs) but retained megakaryocyteerythroid progenitors (MEPs). Consistent with the loss of HSC activity, a 10-fold reduction in erythroid progenitors (mature erythroid burst-forming units [BFUEs]) was observed between e14.5 and e16.5. These data suggest that PU.1 plays an important role in the maintenance or expansion of HSC number in murine fetal liver.


Sexual counselling for patients with cardiovascular disease: protocol for a pilot study of the CHARMS sexual counselling intervention.
Sexual problems are common with cardiovascular disease, and can negatively impact quality of life. To address sexual problems, guidelines have identified the importance of sexual counselling during cardiac rehabilitation, yet this is rarely provided. The Cardiac Health and Relationship Management and Sexuality (CHARMS) intervention aims to improve the provision of sexual counselling in cardiac rehabilitation in Ireland. This is a multicentre pilot study for the CHARMS intervention, a complex, multilevel intervention delivered within hospital-based cardiac rehabilitation programmes. The intervention includes (1) training in sexual counselling for staff, (2) a staff-led patient education and support intervention embedded within the cardiac rehabilitation programme, (3) a patient information booklet and (4) an awareness raising poster. The intervention will be delivered in two randomly selected cardiac rehabilitation centres. In each centre 30 patients will be recruited, and partners will also be invited to participate. Data will be collected from staff and patients/partners at T1 (study entry), T2 (3-month follow-up) and T3 (6-month follow-up). The primary outcome for patients/partners will be scores on the Sexual Self-Perception and Adjustment Questionnaire. Secondary outcomes for patients/partners will include relationship satisfaction; satisfaction with and barriers to sexual counselling in services; sexual activity, functioning and knowledge; physical and psychological well-being. Secondary outcomes for staff will include sexuality-related practice; barriers to sexual counselling; self-ratings of capability, opportunity and motivation; sexual attitudes and beliefs; knowledge of cardiovascular disease and sex. Fidelity of intervention delivery will be assessed using trainer self-reports, researcher-coded audio recordings and exit interviews. Longitudinal feasibility data will be gathered from patients/partners and staff via questionnaires and interviews. This study is approved by the Research Ethics Committee (REC) of the National University of Ireland, Galway. Findings will be disseminated to cardiac rehabilitation staff, patients/partners and relevant policymakers via appropriate publications and presentations.


Normalizing real-time PCR results in routine testing.
Normalization, the process of controlling for normal variation in sampling and testing, can be achieved in real-time PCR assays by converting sample quantification cycles (Cqs) to "efficiency standardized Cqs" (ECqs). We calculated ECqs as E-ΔCq, where E is amplification efficiency and ΔCq is the difference between sample and reference standard Cqs. To apply this approach to a commercial porcine reproductive and respiratory syndrome virus (PRRSV) RT-qPCR assay, we created reference standards by rehydrating and then diluting (1 × 10-4) a PRRSV modified-live vaccine (PRRS MLV; Ingelvac) with serum or oral fluid (OF) to match the sample matrix to be tested. Sample ECqs were calculated using the mean E and reference standard Cq calculated from the 4 reference standards on each plate. Serum (n = 132) and OF (n = 130) samples were collected from each of 12 pigs vaccinated with a PRRSV MLV from -7 to 42 d post-vaccination, tested, and sample Cqs converted to ECqs. Mean plate Es were 1.75-2.6 for serum and 1.7-2.3 for OF. Mean plate reference standard Cqs were 29.1-31.3 for serum and 29.2-31.5 for OFs. Receiver operating characteristic analysis calculated the area under the curve for serum and OF sample ECqs as 0.999 (95% CI: 0.997, 1.000) and 0.947 (0.890, 1.000), respectively. For serum, diagnostic sensitivity and specificity of the commercial PRRSV RT-qPCR assay were estimated as 97.9% and 100% at an ECq cutoff ≥ 0.20, and for OF, 82.6% and 100%, respectively, at an ECq cutoff ≥ 0.45.


[Health inequity among waged workers by employment status].
The purpose of this study was to examine the relationship between the differences in employment status and self assessed health in Korea. We analyzed 4 year follow-up data generated by the Korean Labor and Income Panel Study(KLIPS), which was conducted on 1,207 men and 582 women who had undergone a change in employment status. The study subjects were placed into 1 of the following 4 groups based on their employment history; Non-precarious workers, Precarious to non-precarious workers, Non-precarious to precarious workers and Precarious workers. Logistic regression was then used to examine the relationship between the changes in employment status and self assessed health. When males were considered, self assessed health was better among the precarious to non-precarious workers (OR 1.58, 95% CI=1.57-1.60) and the precarious workers (OR 1.29, 95% CI=1.28-1.30) than in the non-precarious workers, after adjusting for age, socioeconomic status (education level, occupational class, marital status, average equivalent household income and average number of hours worked per week), health behavior (smoking, drinking and exercise) and medical service access (regular medical examination, have chronic disease or hospitalized within 1 year). When female workers were considered, the precarious to non-precarious workers (OR 1.89, 95% CI=1.86-1.92), non-precarious to precarious workers(OR 1.24, 95% CI=1.23-1.26) and precarious workers (OR 1.27, 95% CI=1.25-1.28) all reported poorer health than the non-precarious workers after adjusting for the aforementioned factors. This study showed that changes in employment status were associated with differences in self assessed health among men and women. Specifically, the results of this study showed that a corresponding positive outcome based on self assessed health was greater for employees that changed from precarious to non-precarious jobs and for male employees with precarious jobs, whereas female employees with non-precarious jobs had higher self assessed health. However, additional longitudinal studies on the health effects of employment status should be conducted.


Conducting redox polymers with non-activated charge transport properties.
Non-activated charge transport has been demonstrated in terephthalate-functionalized conducting redox polymers. The transition from a temperature-activated conduction mechanism to a residual scattering mechanism was dependent on the doping level. The latter mechanism is associated with apparent negative activation barriers to charge transport and is generally found in polymer materials with a high degree of order. Crystallographic data, however, suggested a low degree of order in this polymer, indicating the existence of interconnected crystal domains in the predominantly amorphous polymer matrix through which the charge was transported. We have thus shown that the addition of bulky pendant groups to conducting polymers does not prevent efficient charge transport via the residual scattering mechanism with low barriers to charge transport.


A systematic review of psychosocial interventions for adults with autism spectrum disorders.
Individuals with autism spectrum disorders (ASD) spend the majority of their lives as adults, and psychosocial interventions show promise for improving outcomes in this population. This research conducted a systematic review of all peer-review studies evaluating psychosocial interventions for adults with ASD. A total of 1,217 studies were reviewed, only 13 met inclusion criteria. The majority of studies were single case studies or non-randomized controlled trials, and most focused on applied behavior analysis or social cognition training. Effects of psychosocial treatment in adults with ASD were largely positive ranging from d = 0.14-3.59, although the quantity and quality of studies is limited. There is substantial need for the rigorous development and evaluation of psychosocial treatments for adults with ASD.


Certified Nursing Assistants Balancing Family Caregiving Roles: Health Care Utilization Among Double- and Triple-Duty Caregivers.
This study examines how certified nursing assistants (CNAs) balancing family caregiving roles-child care (double-duty child caregivers), elder care (double-duty elder caregivers), and both child and elder care (triple-duty caregivers)-utilize health care services relative to nonfamily caregiving counterparts (formal-only caregivers). A sample of 884 CNAs from the Work, Family and Health Study was drawn on to assess the number of acute care (i.e., emergency room or urgent care facility) and other health care (i.e., outpatient treatment or counseling) visits made during the past 6 months. Double-duty elder and triple-duty caregivers had higher acute care utilization rates than formal-only caregivers. CNAs with and without family caregiving roles had similar rates of other health care visits. CNAs providing informal care for older adults have higher acute care visit rates. Given the increasing need for family caregivers and the vital importance of the health of the nursing workforce for the health of others, future research on how double- and triple-duty caregivers maintain their health amidst constant caregiving should be a priority.


Re-evaluation of sodium aluminium silicate (E 554) and potassium aluminium silicate (E 555) as food additives.
The Panel on Food Additives and Flavourings (FAF) provided a scientific opinion re-evaluating the safety of Sodium aluminium silicate (E 554) and potassium aluminium silicate (E 555) as food additives. The Scientific Committee for Food (SCF) assigned these food additives together with other aluminium-containing food additives a provisional tolerable weekly intake (PTWI) of 7 mg aluminium/kg body weight (bw). In 2008, EFSA established a tolerable weekly intake (TWI) of 1 mg aluminium/kg bw per week. Sodium aluminium silicate was shown in rats to be absorbed to a limited extent at 0.12 ± 0.011%. The Panel considered that potassium aluminium silicate would be absorbed and become systemically available similarly to sodium aluminium silicate. No information on the physicochemical characterisation of sodium aluminium silicate and potassium aluminium silicate when used as food additives has been submitted and only very limited toxicological data were available for sodium aluminium silicate. Exposure to E 554 was calculated based on the reported use levels in food supplements. Exposure to aluminium from this use of E 554 was calculated to exceed the TWI for aluminium. Based on the data provided by interested business operators, the Panel considered that E 555 is not being used as a carrier, but as an inseparable component of 'potassium aluminium silicate-based pearlescent pigments'. The Panel calculated the regulatory maximum exposure to E 555 as a carrier for titanium dioxide (E 171) and iron oxides and hydroxides (E 172). Exposure to aluminium from this single use at the maximum permitted level could theoretically far exceed the TWI. Considering that only very limited toxicological data and insufficient information on the physicochemical characterisation of both food additives were available, the Panel concluded that the safety of sodium aluminium silicate (E 554) and potassium aluminium silicate (E 555) could not be assessed.


SU-E-T-614: Dose-Reactive Methods in Adaptive Robust Radiation Therapy for Lung Cancer.
To perform adaptive radiation therapy treatments for lung cancer using IMRT; to show that adjusting the target dose after each fraction, in order to'react' to errors in the dose delivered in prior fractions, can lead to significant changes in the daily tumor dose over the treatment. Before the start of treatment, the beamlet intensities are optimized to deliver the current target dose distribution for a target set of breathing patterns at minimal healthy tissue dose. In each fraction, the current beamlet intensities are delivered, and the patient's breathing pattern is measured. The breathing pattern set is updated using the breathing pattern, and the target dose distribution is updated to account for dose errors realized in the previous fraction. The beamlet intensities are then re-optimized for the updated dose distribution and uncertainty set, to be used in the next fraction. This process continues until the end of the treatment. We consider three types of updates to the target dose distribution: the reactive± update, which responds to both under and overdose; the reactive- update, which responds only to underdose; and the reactive+ update, which responds only to overdose. On breathing pattern sequences obtained from real patients, dose-reactive methods result in final dose performance comparable to non-dose-reactive methods. However, as the treatment progresses, the reactive± update results in growing daily underdose and overdose, the reactive- update results in growing daily overdose, and the reactive+ update results in growing daily underdose. In contrast, non-reactive methods have stable or decreasing tumor underdose and overdose. By incorporating dose-reaction, the final tumor dose distribution can be made to conform closely to the target dose distribution, but at the cost of increasing tumor underdose and/or overdose. This increasing heterogeneity may have implications for the biological effectiveness of treatments obtained by dose-reaction.


Relapse to heroin-seeking in rats under opioid maintenance: the effects of stress, heroin priming, and withdrawal.
It is widely believed that opioid withdrawal symptoms contribute to relapse to opioid use, but relapse is highly probable in experienced users even after prolonged abstinence and during opioid maintenance therapy. We have found using an animal model of relapse, the reinstatement procedure, that the two events that reliably reinstate heroin-seeking behavior are reexposure to heroin, and brief exposure to footshock stress. Contrary to expectation, opioid antagonist-induced withdrawal does not reinstate heroin-seeking. We now report on reinstatement of heroin-seeking in rats trained to self-administer heroin and subsequently exposed to a maintenance dose of heroin via minipump and allowed to self-administer saline. With the minipump in, naloxone-induced withdrawal did not reinstate drug-seeking, a priming injection on heroin was only mildly effective, and footshock was highly effective. Twenty-four hours after removal of the minipump (spontaneous withdrawal), animals reinitiated heroin-seeking and, subsequently, both heroin and footshock reinstated heroin-seeking. In summary, brief exposure to stress reinstated heroin-seeking in both heroin-maintained and withdrawn animals. The heroin prime reliably reinstated drug-seeking only in the absence of the minipump; opioid "withdrawal," as such, did not reinstate drug-seeking behavior. Naloxone given to heroin-maintained animals induced withdrawal symptoms, caused a mild depression in the levels of dopamine and its metabolites in the nucleus accumbens septi (NAS), but did not reinstate drug-seeking. Reinstatement of heroin-seeking during spontaneous withdrawal was not accompanied by reductions in basal dopamine and its metabolites in NAS.


Integration of sample preparation and analysis into an optofluidic chip for multi-target disease detection.
Detection of molecular biomarkers with high specificity and sensitivity from biological samples requires both sophisticated sample preparation and subsequent analysis. These tasks are often carried out on separate platforms which increases required sample volumes and the risk of errors, sample loss, and contamination. Here, we present an optofluidic platform which combines an optical detection section with single nucleic acid strand sensitivity, and a sample processing unit capable of on-chip, specific extraction and labeling of nucleic acid and protein targets in complex biological matrices. First, on-chip labeling and detection of individual lambda DNA molecules down to concentrations of 8 fM is demonstrated. Subsequently, we demonstrate the simultaneous capture, fluorescence tagging and detection of both Zika specific nucleic acid and NS-1 protein targets in both buffer and human serum. We show that the dual DNA and protein assay allows for successful differentiation and diagnosis of Zika against cross-reacting species like dengue.


The Genetics of Neuropathic Pain from Model Organisms to Clinical Application.
Neuropathic pain (NeuP) arises due to injury of the somatosensory nervous system and is both common and disabling, rendering an urgent need for non-addictive, effective new therapies. Given the high evolutionary conservation of pain, investigative approaches from Drosophila mutagenesis to human Mendelian genetics have aided our understanding of the maladaptive plasticity underlying NeuP. Successes include the identification of ion channel variants causing hyper-excitability and the importance of neuro-immune signaling. Recent developments encompass improved sensory phenotyping in animal models and patients, brain imaging, and electrophysiology-based pain biomarkers, the collection of large well-phenotyped population cohorts, neurons derived from patient stem cells, and high-precision CRISPR generated genetic editing. We will discuss how to harness these resources to understand the pathophysiological drivers of NeuP, define its relationship with comorbidities such as anxiety, depression, and sleep disorders, and explore how to apply these findings to the prediction, diagnosis, and treatment of NeuP in the clinic.


GNAS mutations in Pseudohypoparathyroidism type 1a and related disorders.
Pseudohypoparathyroidism type 1a (PHP1a) is characterized by hypocalcaemia and hyperphosphatemia due to parathyroid hormone resistance, in association with the features of Albright's hereditary osteodystrophy (AHO). PHP1a is caused by maternally inherited inactivating mutations of Gs-alpha, which is encoded by a complex imprinted locus termed GNAS. Paternally inherited mutations can lead either to pseudopseudohypoparathyroidism (PPHP) characterized by AHO alone, or to progressive osseous heteroplasia (POH), characterized by severe heterotopic ossification. The clinical aspects and molecular genetics of PHP1a and its related disorders are reviewed together with the 343 kindreds with Gs-alpha germline mutations reported so far in the literature. These 343 (176 different) mutations are scattered throughout the 13 exons that encode Gs-alpha and consist of 44.9% frameshift, 28.0% missense, 14.0% nonsense, and 9.0% splice-site mutations, 3.2% in-frame deletions or insertions, and 0.9% whole or partial gene deletions. Frameshift and other highly disruptive mutations were more frequent in the reported 37 POH kindreds than in PHP1a/PPHP kindreds (97.3% vs. 68.7%, P < 0.0001). This mutation update and respective genotype-phenotype data may be of use for diagnostic and research purposes and contribute to a better understanding of these complex disorders.


OX40 expression in neutrophils promotes hepatic ischemia/reperfusion injury.
Neutrophils play critical roles during the initial phase of hepatic ischemia/reperfusion injury (HIRI). However, the regulation of neutrophil activation, infiltration, and proinflammatory cytokine secretion has not been fully elucidated. In this study, we revealed that OX40 was expressed by neutrophils, its expression in neutrophils was time-dependently upregulated following HIRI, and Ox40 knockout markedly alleviated liver injury. Compared with wild-type neutrophils, the adoptive transfer of Ox40-/- neutrophils decreased HIRI in neutrophil-depleted Rag2/Il2rg-/- or Ox40-/- mice. Moreover, consistently, the in vitro experiments showed that Ox40 not only prolonged neutrophil survival but also promoted proinflammatory cytokines, ROS production, and even neutrophil chemotaxis. Further investigation demonstrated that the knockout of Ox40 in neutrophils inhibited NF-κB signaling via the TRAF1/2/4 and IKKα/IKKβ/IκBα pathways. OX40L and OX86 stimulation could enhance neutrophil activation and survival in vitro and in vivo. In conclusion, our study provides a new understanding of OX40, which is expressed not only in adaptive immune cells but also in innate immune cells, i.e., neutrophils, contributing to the activation and survival of neutrophils. These findings provide a novel potential therapeutic target for the prevention of HIRI during liver transplantation or hepatic surgery.


Graph Drawing Aesthetics-Created by Users, Not Algorithms.
Prior empirical work on layout aesthetics for graph drawing algorithms has concentrated on the interpretation of existing graph drawings. We report on experiments which focus on the creation and layout of graph drawings: participants were asked to draw graphs based on adjacency lists, and to lay them out "nicely." Two interaction methods were used for creating the drawings: a sketch interface which allows for easy, natural hand movements, and a formal point-and-click interface similar to a typical graph editing system. We find, in common with many other studies, that removing edge crossings is the most significant aesthetic, but also discover that aligning nodes and edges to an underlying grid is important. We observe that the aesthetics favored by participants during creation of a graph drawing are often not evident in the final product and that the participants did not make a clear distinction between the processes of creation and layout. Our results suggest that graph drawing systems should integrate automatic layout with the user's manual editing process, and provide facilities to support grid-based graph creation.


Support vector machines for detecting age-related changes in running kinematics.
Age-related changes in running kinematics have been reported in the literature using classical inferential statistics. However, this approach has been hampered by the increased number of biomechanical gait variables reported and subsequently the lack of differences presented in these studies. Data mining techniques have been applied in recent biomedical studies to solve this problem using a more general approach. In the present work, we re-analyzed lower extremity running kinematic data of 17 young and 17 elderly male runners using the Support Vector Machine (SVM) classification approach. In total, 31 kinematic variables were extracted to train the classification algorithm and test the generalized performance. The results revealed different accuracy rates across three different kernel methods adopted in the classifier, with the linear kernel performing the best. A subsequent forward feature selection algorithm demonstrated that with only six features, the linear kernel SVM achieved 100% classification performance rate, showing that these features provided powerful combined information to distinguish age groups. The results of the present work demonstrate potential in applying this approach to improve knowledge about the age-related differences in running gait biomechanics and encourages the use of the SVM in other clinical contexts.


A Multisite Preregistered Paradigmatic Test of the Ego-Depletion Effect.
We conducted a preregistered multilaboratory project (k = 36; N = 3,531) to assess the size and robustness of ego-depletion effects using a novel replication method, termed the paradigmatic replication approach. Each laboratory implemented one of two procedures that was intended to manipulate self-control and tested performance on a subsequent measure of self-control. Confirmatory tests found a nonsignificant result (d = 0.06). Confirmatory Bayesian meta-analyses using an informed-prior hypothesis (δ = 0.30, SD = 0.15) found that the data were 4 times more likely under the null than the alternative hypothesis. Hence, preregistered analyses did not find evidence for a depletion effect. Exploratory analyses on the full sample (i.e., ignoring exclusion criteria) found a statistically significant effect (d = 0.08); Bayesian analyses showed that the data were about equally likely under the null and informed-prior hypotheses. Exploratory moderator tests suggested that the depletion effect was larger for participants who reported more fatigue but was not moderated by trait self-control, willpower beliefs, or action orientation.


Production and properties of hemicellulases by a Thermomyces lanuginosus strain.
Thermophilic fungi producing extremely high beta-xylanase and their associated hemicellulases have attracted considerable attention because of potential industrial applications. Thermomyces lanuginosus strain SSBP isolated from soil, produced beta-xylanase activity of 59 600 nkat ml-1 when cultivated on a medium containing corn cobs as substrate and yeast extract as nitrogen source. Lower beta-xylanase activities were produced after growth on other xylan substrates, sugars and soluble starch. Other hemicellulases were produced extracellularly at significantly lower levels than the beta-xylanase activity produced on corn cobs. No cellulase activity was observed. The optimal conditions for beta-xylanase production were 50 degrees C and pH 6.5, whereas 70 degrees C and between pH 5. 5 and 9.5 were optimal for beta-xylanase activity. The temperature optima for other hemicellulases were less than the xylanase with the exception of beta-mannosidase. The pH optima of the other hemicellulases were between 5.0 and 6.5. Xylanase was stable up to 70 degrees C and between pH 5.5 and 9.0 for 30 min whereas the other hemicellulase were less stable. These results suggest that the most suitable conditions for hydrolysis of hemicellulose by these enzymes would be at 50 degrees C and pH 6.0.


Positive CD63 Basophil Activation Tests Are Common in Children with Chronic Spontaneous Urticaria and Linked to High Disease Activity.
The basophil activation test (BAT) using CD63 expression is a sensitive and specific tool for the diagnostic workup of autoimmune chronic spontaneous urticaria (CSU). The definition of a positive BAT is directly dependent on the reference range and the cutoff values established in control populations. As of now, the pediatric reference range and cutoff values of the CD63 BAT remain to be established. In this study, we analyzed CD63 expression in 80 children (1-17 years old) without chronic urticaria (i.e., controls) and compared the values to those of a pediatric cohort of 105 CSU patients and 23 physical urticaria (PU) patients. Based on the log-normal distribution of CD63 values in control subjects, the reference range and the cutoff for positive CD63 BAT values was established to be 1.2-1.8% (95% CI) and 1.8%, respectively. Children with CSU showed significantly elevated and significantly increased BAT values compared to healthy controls (Wilcoxon rank test p value <0.001). In contrast, no difference was found between BAT results in controls and PU patients. In pediatric CSU patients, a higher disease activity was associated with higher BAT values. Our study provides, for the first time, reference and cutoff values for the CD63 BAT in children. Our findings show that positive CD63 BAT are common in children with CSU and linked to a high disease activity.


Migration and Development: A Theoretical Perspective.
The debate on migration and development has swung back and forth like a pendulum, from developmentalist optimism in the 1950s and 1960s, to neo-Marxist pessimism over the 1970s and 1980s, towards more optimistic views in the 1990s and 2000s. This paper argues how such discursive shifts in the migration and development debate should be primarily seen as part of more general paradigm shifts in social and development theory. However, the classical opposition between pessimistic and optimistic views is challenged by empirical evidence pointing to the heterogeneity of migration impacts. By integrating and amending insights from the new economics of labor migration, livelihood perspectives in development studies and transnational perspectives in migration studies - which share several though as yet unobserved conceptual parallels - this paper elaborates the contours of a conceptual framework that simultaneously integrates agency and structure perspectives and is therefore able to account for the heterogeneous nature of migration-development interactions. The resulting perspective reveals the naivety of recent views celebrating migration as self-help development "from below". These views are largely ideologically driven and shift the attention away from structural constraints and the vital role of states in shaping favorable conditions for positive development impacts of migration to occur.


Commissioning a CT-compatible LDR tandem and ovoid applicator using Monte Carlo calculation and 3D dosimetry.
To determine the geometric and dose attenuation characteristics of a new commercially available CT-compatible LDR tandem and ovoid (T&O) applicator using Monte Carlo calculation and 3D dosimetry. For geometric characterization, we quantified physical dimensions and investigated a systematic difference found to exist between nominal ovoid angle and the angle at which the afterloading buckets fall within the ovoid. For dosimetric characterization, we determined source attenuation through asymmetric gold shielding in the buckets using Monte Carlo simulations and 3D dosimetry. Monte Carlo code MCNP5 was used to simulate 1.5 × 10(9) photon histories from a (137)Cs source placed in the bucket to achieve statistical uncertainty of 1% at a 6 cm distance. For 3D dosimetry, the distribution about an unshielded source was first measured to evaluate the system for (137)Cs, after which the distribution was measured about sources placed in each bucket. Cylindrical PRESAGE(®) dosimeters (9.5 cm diameter, 9.2 cm height) with a central channel bored for source placement were supplied by Heuris Inc. The dosimeters were scanned with the Duke Large field of view Optical CT-Scanner before and after delivering a nominal dose at 1 cm of 5-8 Gy. During irradiation the dosimeter was placed in a water phantom to provide backscatter. Optical CT scan time lasted 15 min during which 720 projections were acquired at 0.5° increments, and a 3D distribution was reconstructed with a (0.05 cm)(3) isotropic voxel size. The distributions about the buckets were used to calculate a 3D distribution of transmission rate through the bucket, which was applied to a clinical CT-based T&O implant plan. The systematic difference in bucket angle relative to the nominal ovoid angle (105°) was 3.1°-4.7°. A systematic difference in bucket angle of 1°, 5°, and 10° caused a 1% ± 0.1%, 1.7% ± 0.4%, and 2.6% ± 0.7% increase in rectal dose, respectively, with smaller effect to dose to Point A, bladder, sigmoid, and bowel. For 3D dosimetry, 90.6% of voxels had a 3D γ-index (criteria = 0.1 cm, 3% local signal) below 1.0 when comparing measured and expected dose about the unshielded source. Dose transmission through the gold shielding at a radial distance of 1 cm was 85.9% ± 0.2%, 83.4% ± 0.7%, and 82.5% ± 2.2% for Monte Carlo, and measurement for left and right buckets, respectively. Dose transmission was lowest at oblique angles from the bucket with a minimum of 56.7% ± 0.8%, 65.6% ± 1.7%, and 57.5% ± 1.6%, respectively. For a clinical T&O plan, attenuation from the buckets leads to a decrease in average Point A dose of ∼3.2% and decrease in D(2cc) to bladder, rectum, bowel, and sigmoid of 5%, 18%, 6%, and 12%, respectively. Differences between dummy and afterloading bucket position in the ovoids is minor compared to effects from asymmetric ovoid shielding, for which rectal dose is most affected. 3D dosimetry can fulfill a novel role in verifying Monte Carlo calculations of complex dose distributions as are common about brachytherapy sources and applicators.


Direct visualization of cardiac radiofrequency ablation lesions.
Effective ablation of atrial fibrillation and other cardiac arrhythmias requires precise catheter navigation and controlled delivery of energy to cardiac tissue. In this study, we summarize our initial experience using a fiber optic direct visualization catheter to evaluate and guide placement of endocardial radiofrequency (RF) ablation lesions. RF lesions were created in cadaveric porcine hearts and examined in a blood-filled field using a direct visualization catheter. Direct visualization of RF lesions was repeated in vivo using an ovine model. Lesions and interlesion gaps were clearly identifiable using the direct visualization catheter. It was possible to place lesions in proximity to anatomical landmarks and in relation to one another. Catheter-generated images correlated well with lesion appearance on gross examination. Direct catheter-based visualization is a feasible technique for guiding RF lesion placement, estimating lesion size, and identifying interlesion gaps. Future work is needed to correlate surface appearance with transmurality and electrical isolation.


Genomic Approaches to Understanding Response and Resistance to Immunotherapy.
Immunotherapy has led to a paradigm shift in the treatment of some malignancies, providing long-term, durable responses for patients with advanced cancers. However, such therapy has benefited only a subset of patients, with some patients failing to respond to treatment at all and others achieving a limited response followed by tumor progression. Understanding factors contributing to an effective response and further elucidating mechanisms of resistance will be crucial as these therapies are applied more broadly. Genomics-based approaches have significantly advanced the study of response and resistance to immunotherapy in general, and to immune checkpoint blockade more specifically. Here, we review how genomic and transcriptomic approaches have identified both somatic and germline positive correlates of response, including high mutational/neoantigen load and low intratumoral heterogeneity, among others. The genomic analysis of resistant tumors has additionally identified crucial factors involved in resistance to immune checkpoint blockade, including loss of PTEN and upregulation of other immune checkpoints. Overall, the continued use of genomic techniques at the point of care, combined with appropriate functional studies, would ideally lead to a better understanding of why certain patients respond to immune-based therapies, allowing clinicians to identify the subset of patients likely to benefit from such therapy, and potentially providing insight into how other therapies may be added in combination to increase the number of patients who may benefit from immunotherapy. Clin Cancer Res; 22(23); 5642-50. ©2016 AACR.


Transcriptome analysis of pancreatic cancer reveals a tumor suppressor function for HNF1A.
Pancreatic ductal adenocarcinoma (PDAC) is driven by the accumulation of somatic mutations, epigenetic modifications and changes in the micro-environment. New approaches to investigating disruptions of gene expression networks promise to uncover key regulators and pathways in carcinogenesis. We performed messenger RNA-sequencing in pancreatic normal (n = 10) and tumor (n = 8) derived tissue samples, as well as in pancreatic cancer cell lines (n = 9), to determine differential gene expression (DE) patterns. Sub-network enrichment analyses identified HNF1A as the regulator of the most significantly and consistently dysregulated expression sub-network in pancreatic tumor tissues and cells (median P = 7.56×10(-7), median rank = 1, range = 1-25). To explore the effects of HNF1A expression in pancreatic tumor-derived cells, we generated stable HNF1A-inducible clones in two pancreatic cancer cell lines (PANC-1 and MIA PaCa-2) and observed growth inhibition (5.3-fold, P = 4.5×10(-5) for MIA PaCa-2 clones; 7.2-fold, P = 2.2×10(-5) for PANC-1 clones), and a G0/G1 cell cycle arrest and apoptosis upon induction. These effects correlated with HNF1A-induced down-regulation of 51 of 84 cell cycle genes (e.g. E2F1, CDK2, CDK4, MCM2/3/4/5, SKP2 and CCND1), decreased expression of anti-apoptotic genes (e.g. BIRC2/5/6 and AKT) and increased expression of pro-apoptotic genes (e.g. CASP4/9/10 and APAF1). In light of the established role of HNF1A in the regulation of pancreatic development and homeostasis, our data suggest that it also functions as an important tumor suppressor in the pancreas.


Using Bayesian adaptive designs to improve phase III trials: a respiratory care example.
Bayesian adaptive designs can improve the efficiency of trials, and lead to trials that can produce high quality evidence more quickly, with fewer patients and lower costs than traditional methods. The aim of this work was to determine how Bayesian adaptive designs can be constructed for phase III clinical trials in critical care, and to assess the influence that Bayesian designs would have on trial efficiency and study results. We re-designed the High Frequency OSCillation in Acute Respiratory distress syndrome (OSCAR) trial using Bayesian adaptive design methods, to allow for the possibility of early stopping for success or futility. We constructed several alternative designs and studied their operating characteristics via simulation. We then performed virtual re-executions by applying the Bayesian adaptive designs using the OSCAR data to demonstrate the practical applicability of the designs. We constructed five alternative Bayesian adaptive designs and identified a preferred design based on the simulated operating characteristics, which had similar power to the original design but recruited fewer patients on average. The virtual re-executions showed the Bayesian sequential approach and original OSCAR trial yielded similar trial conclusions. However, using a Bayesian sequential design could have led to a reduced sample size and earlier completion of the trial. Using the OSCAR trial as an example, this case study found that Bayesian adaptive designs can be constructed for phase III critical care trials. If the OSCAR trial had been run using one of the proposed Bayesian adaptive designs, it would have terminated at a smaller sample size with fewer deaths in the trial, whilst reaching the same conclusions. We recommend the wider use of Bayesian adaptive approaches in phase III clinical trials. OSCAR Trial registration ISRCTN, ISRCTN10416500 . Retrospectively registered 13 June 2007.


Deriving motor primitives through action segmentation.
The purpose of the present experiment is to further understand the effect of levels of processing (top-down vs. bottom-up) on the perception of movement kinematics and primitives for grasping actions in order to gain insight into possible primitives used by the mirror system. In the present study, we investigated the potential of identifying such primitives using an action segmentation task. Specifically, we investigated whether or not segmentation was driven primarily by the kinematics of the action, as opposed to high-level top-down information about the action and the object used in the action. Participants in the experiment were shown 12 point-light movies of object-centered hand/arm actions that were either presented in their canonical orientation together with the object in question (top-down condition) or upside down (inverted) without information about the object (bottom-up condition). The results show that (1) despite impaired high-level action recognition for the inverted actions participants were able to reliably segment the actions according to lower-level kinematic variables, (2) segmentation behavior in both groups was significantly related to the kinematic variables of change in direction, velocity, and acceleration of the wrist (thumb and finger tips) for most of the included actions. This indicates that top-down activation of an action representation leads to similar segmentation behavior for hand/arm actions compared to bottom-up, or local, visual processing when performing a fairly unconstrained segmentation task. Motor primitives as parts of more complex actions may therefore be reliably derived through visual segmentation based on movement kinematics.


Associations of Various Health-Ratings with Geriatric Giants, Mortality and Life Satisfaction in Older People.
Self-rated health is routinely used in research and practise among general populations. Older people, however, seem to change their health perceptions. To accurately understand these changed perceptions we therefore need to study the correlates of older people's self-ratings. We examined self-rated, nurse-rated and physician-rated health's association with common disabilities in older people (the geriatric giants), mortality hazard and life satisfaction. For this, we used an age-representative population of 501 participant aged 85 from a middle-sized city in the Netherlands: the Leiden 85-plus Study. Participants with severe cognitive dysfunction were excluded. Participants themselves provided health ratings, as well as a visiting physician and a research nurse. Visual acuity, hearing loss, mobility, stability, urinal and faecal incontinence, cognitive function and mood (depressive symptoms) were included as geriatric giants. Participants provided a score for life satisfaction and were followed up for vital status. Concordance of self-rated health with physician-rated (k = .3 [.0]) and nurse-rated health (k = .2 [.0]) was low. All three ratings were associated with the geriatric giants except for hearing loss (all p < 0.001). Associations were equal in strength, except for depressive symptoms, which showed a stronger association with self-rated health (.8 [.1] versus .4 [.1]). Self-rated health predicted mortality less well than the other ratings. Self-rated health related stronger to life satisfaction than physician's and nurse's ratings. We conclude that professionals' health ratings are more reflective of physical health whereas self-rated health reflects more the older person's mental health, but all three health ratings are useful in research.


Search for pair production of first-generation scalar leptoquarks in pp collisions at √s = 7 TeV.
A search for pair production of first-generation scalar leptoquarks is performed in the final state containing two electrons and two jets using proton-proton collision data at √s = 7 TeV. The data sample used corresponds to an integrated luminosity of 33 pb⁻¹ collected with the CMS detector at the CERN LHC. The number of observed events is in good agreement with the predictions for the standard model background processes, and an upper limit is set on the leptoquark pair production cross section times β² as a function of the leptoquark mass, where β is the branching fraction of the leptoquark decay to an electron and a quark. A 95% confidence level lower limit is set on the mass of a first-generation scalar leptoquark at 384 GeV for β = 1, which is the most stringent direct limit to date.


RIM controls homeostatic plasticity through modulation of the readily-releasable vesicle pool.
Rab3 interacting molecules (RIMs) are evolutionarily conserved scaffolding proteins that are located at presynaptic active zones. In the mammalian nervous system, RIMs have two major activities that contribute to the fidelity of baseline synaptic transmission: they concentrate calcium channels at the active zone and facilitate synaptic vesicle docking/priming. Here we confirm that RIM has an evolutionarily conserved function at the Drosophila neuromuscular junction and then define a novel role for RIM during homeostatic synaptic plasticity. We show that loss of RIM disrupts baseline vesicle release, diminishes presynaptic calcium influx, and diminishes the size of the readily-releasable pool (RRP) of synaptic vesicles, consistent with known activities of RIM. However, loss of RIM also completely blocks the homeostatic enhancement of presynaptic neurotransmitter release that normally occurs after inhibition of postsynaptic glutamate receptors, a process termed synaptic homeostasis. It is established that synaptic homeostasis requires enhanced presynaptic calcium influx as a mechanism to potentiate vesicle release. However, despite a defect in baseline calcium influx in rim mutants, the homeostatic modulation of calcium influx proceeds normally. Synaptic homeostasis is also correlated with an increase in the size of the RRP of synaptic vesicles, although the mechanism remains unknown. Here we demonstrate that the homeostatic modulation of the RRP is blocked in the rim mutant background. Therefore, RIM-dependent modulation of the RRP is a required step during homeostatic plasticity. By extension, homeostatic plasticity appears to require two genetically separable processes, the enhancement of presynaptic calcium influx and a RIM-dependent modulation of the RRP.


Suppression of eIF2α kinases alleviates Alzheimer's disease-related plasticity and memory deficits.
Expression of long-lasting synaptic plasticity and long-term memory requires protein synthesis, which can be repressed by phosphorylation of eukaryotic initiation factor 2 α-subunit (eIF2α). Elevated phosphorylation of eIF2α has been observed in the brains of Alzheimer's disease patients and Alzheimer's disease model mice. Therefore, we tested whether suppressing eIF2α kinases could alleviate synaptic plasticity and memory deficits in Alzheimer's disease model mice. Genetic deletion of eIF2α kinase PERK prevented enhanced phosphorylation of eIF2α and deficits in protein synthesis, synaptic plasticity and spatial memory in mice that express familial Alzheimer's disease-related mutations in APP and PSEN1. Similarly, deletion of another eIF2α kinase, GCN2, prevented impairments of synaptic plasticity and defects in spatial memory exhibited by the Alzheimer's disease model mice. Our findings implicate aberrant eIF2α phosphorylation as a previously unidentified molecular mechanism underlying Alzheimer's disease-related synaptic pathophysioloy and memory dysfunction and suggest that PERK and GCN2 are potential therapeutic targets for treatment of individuals with Alzheimer's disease.


Visual hallucinations in Parkinson's disease: theoretical models.
One of the most challenging tasks in neuroscience is to be able to meaningfully connect information across the different levels of investigation, from molecular or structural biology to the resulting behavior and cognition. Visual hallucinations are a frequent occurrence in Parkinson's disease and significantly contribute to the burden of the disease. Because of the widespread pathological processes implicated in visual hallucinations in Parkinson's disease, a final common mechanism that explains their manifestation will require an integrative approach, in which consideration is taken across all complementary levels of analysis. This review considers the leading hypothetical frameworks for visual hallucinations in Parkinson's disease, summarizing the key aspects of each in an attempt to highlight the aspects of the condition that such a unifying hypothesis must explain. These competing hypotheses include implications of dream imagery intrusion, deficits in reality monitoring, and impairments in visual perception and attention.


A methodology for ecosystem-scale modeling of selenium.
The main route of exposure for selenium (Se) is dietary, yet regulations lack biologically based protocols for evaluations of risk. We propose here an ecosystem-scale model that conceptualizes and quantifies the variables that determine how Se is processed from water through diet to predators. This approach uses biogeochemical and physiological factors from laboratory and field studies and considers loading, speciation, transformation to particulate material, bioavailability, bioaccumulation in invertebrates, and trophic transfer to predators. Validation of the model is through data sets from 29 historic and recent field case studies of Se-exposed sites. The model links Se concentrations across media (water, particulate, tissue of different food web species). It can be used to forecast toxicity under different management or regulatory proposals or as a methodology for translating a fish-tissue (or other predator tissue) Se concentration guideline to a dissolved Se concentration. The model illustrates some critical aspects of implementing a tissue criterion: 1) the choice of fish species determines the food web through which Se should be modeled, 2) the choice of food web is critical because the particulate material to prey kinetics of bioaccumulation differs widely among invertebrates, 3) the characterization of the type and phase of particulate material is important to quantifying Se exposure to prey through the base of the food web, and 4) the metric describing partitioning between particulate material and dissolved Se concentrations allows determination of a site-specific dissolved Se concentration that would be responsible for that fish body burden in the specific environment. The linked approach illustrates that environmentally safe dissolved Se concentrations will differ among ecosystems depending on the ecological pathways and biogeochemical conditions in that system. Uncertainties and model sensitivities can be directly illustrated by varying exposure scenarios based on site-specific knowledge. The model can also be used to facilitate site-specific regulation and to present generic comparisons to illustrate limitations imposed by ecosystem setting and inhabitants. Used optimally, the model provides a tool for framing a site-specific ecological problem or occurrence of Se exposure, quantify exposure within that ecosystem, and narrow uncertainties about how to protect it by understanding the specifics of the underlying system ecology, biogeochemistry, and hydrology.


Development of clinical practice guidelines for supportive care in childhood cancer--prioritization of topics using a Delphi approach.
Currently, very few guidelines for supportive care for children with cancer exist. In the Netherlands, nationwide guidelines are over 10 years old and mostly based on expert opinion. Consequently, there is growing support and need for clinical practice guidelines (CPGs), which ought to be developed with a well-defined methodology and include a systematic search of literature, evidence summaries, and a transparent description of the decision process for the final recommendations. Development of CPGs is time consuming; therefore, it is important to prioritize topics for which there is the greatest clinical demand. This study aims to prioritize childhood cancer supportive care topics for development of CPGs. A Delphi survey consisting of two rounds was conducted to prioritize relevant childhood cancer supportive care topics for the development of CPGs. A group of experts comprising 15 pediatric oncologists, 15 pediatric oncology nurses, and 15 general pediatricians involved in care for childhood cancer patients were invited to participate. All relevant supportive care topics in childhood cancer were rated. In both rounds, 36 panellists (82%) responded. Agreement between panellists was very good, with an intraclass correlation coefficient of 0.918 (95% confidence interval (CI) = 0.849-0.966, p < 0.001) in round 2. The ten topics with the highest score in the final round were infection, sepsis, febrile neutropenia, pain, nausea/vomiting, restrictions in daily life and activities, palliative care, procedural sedation, terminal care, and oral mucositis. We successfully used a Delphi survey to prioritize childhood cancer supportive care topics for the development of CPGs. This is a first step towards uniform and evidence-based Dutch guidelines in supportive care in childhood cancer. Even though performed nationally, we believe that this study can also be regarded as an example starting point for international development of CPGs in the field of supportive care in cancer or any other field for that matter.


Supercomputing enabling exhaustive statistical analysis of genome wide association study data: Preliminary results.
Most published GWAS do not examine SNP interactions due to the high computational complexity of computing p-values for the interaction terms. Our aim is to utilize supercomputing resources to apply complex statistical techniques to the world's accumulating GWAS, epidemiology, survival and pathology data to uncover more information about genetic and environmental risk, biology and aetiology. We performed the Bayesian Posterior Probability test on a pseudo data set with 500,000 single nucleotide polymorphism and 100 samples as proof of principle. We carried out strong scaling simulations on 2 to 4,096 processing cores with factor 2 increments in partition size. On two processing cores, the run time is 317h, i.e. almost two weeks, compared to less than 10 minutes on 4,096 processing cores. The speedup factor is 2,020 that is very close to the theoretical value of 2,048. This work demonstrates the feasibility of performing exhaustive higher order analysis of GWAS studies using independence testing for contingency tables. We are now in a position to employ supercomputers with hundreds of thousands of threads for higher order analysis of GWAS data using complex statistics.


Effects of exposure to bodies of different sizes on perception of and satisfaction with own body size: two randomized studies.
Body dissatisfaction is prevalent among women and associated with subsequent obesity and eating disorders. Exposure to images of bodies of different sizes has been suggested to change the perception of 'normal' body size in others. We tested whether exposure to different-sized (otherwise identical) bodies changes perception of own and others' body size, satisfaction with body size and amount of chocolate consumed. In Study 1, 90 18-25-year-old women with normal BMI were randomized into one of three groups to complete a 15 min two-back task using photographs of women either of 'normal weight' (Body Mass Index (BMI) 22-23 kg m-2), or altered to appear either under- or over-weight. Study 2 was identical except the 96 participants had high baseline body dissatisfaction and were followed up after 24 h. We also conducted a mega-analysis combining both studies. Participants rated size of others' bodies, own size, and satisfaction with size pre- and post-task. Post-task ratings were compared between groups, adjusting for pre-task ratings. Participants exposed to over- or normal-weight images subsequently perceived others' bodies as smaller, in comparison to those shown underweight bodies (p < 0.001). They also perceived their own bodies as smaller (Study 1, p = 0.073; Study 2, p = 0.018; mega-analysis, p = 0.001), and felt more satisfied with their size (Study 1, p = 0.046; Study 2, p = 0.004; mega-analysis, p = 0.006). There were no differences in chocolate consumption. This study suggests that a move towards using images of women with a BMI in the healthy range in the media may help to reduce body dissatisfaction, and the associated risk of eating disorders.


High thrombotic risk increases adverse clinical events up to 5 years after acute myocardial infarction. A nationwide retrospective cohort study.
The risk of recurrent events among survivors of acute myocardial infarction (AMI) is understudied. The aim of this analysis was to investigate the role of residual high thrombotic risk (HTR) as a predictor of recurrent in-hospital events after AMI. This retrospective cohort study included 186,646 patients admitted with AMI from 2009 to 2010 in all Italian hospitals who were alive 30 days after the index event. HTR was defined as at least one of the following in the 5 years preceding AMI: previous myocardial infarction, ischemic stroke/other vascular disease, type 2 diabetes mellitus, renal failure. Risk adjustment was performed in all multivariate survival analyses. Rates of major cardiac and cerebrovascular events (MACCE) within the following 5 years were calculated in both patients without fatal readmissions at 30 days and in those free from in-hospital MACCE at 1 year from the index hospitalization. The overall 5-year risk of MACCE was higher in patients with HTR than in those without HTR, in both survivors at 30 days [hazard ratio (HR), 1.49; 95% confidence interval (CI), 1.45-1.52; p<0.0001] and in those free from MACCE at 1 year (HR, 1.46; 95% CI, 1.41-1.51; p<0.0001). The risk of recurrent MACCE increased in the first 18 months after AMI (HR, 1.49) and then remained stable over 5 years. The risk of MACCE after an AMI endures over 5 years in patients with HTR. This is also true for patients who did not have any new cardiovascular event in the first year after an AMI. All patients with HTR should be identified and addressed to intensive preventive care strategies.


Reprogramming towards pluripotency requires AID-dependent DNA demethylation.
Reprogramming of somatic cell nuclei to yield induced pluripotent stem (iPS) cells makes possible derivation of patient-specific stem cells for regenerative medicine. However, iPS cell generation is asynchronous and slow (2-3 weeks), the frequency is low (<0.1%), and DNA demethylation constitutes a bottleneck. To determine regulatory mechanisms involved in reprogramming, we generated interspecies heterokaryons (fused mouse embryonic stem (ES) cells and human fibroblasts) that induce reprogramming synchronously, frequently and fast. Here we show that reprogramming towards pluripotency in single heterokaryons is initiated without cell division or DNA replication, rapidly (1 day) and efficiently (70%). Short interfering RNA (siRNA)-mediated knockdown showed that activation-induced cytidine deaminase (AID, also known as AICDA) is required for promoter demethylation and induction of OCT4 (also known as POU5F1) and NANOG gene expression. AID protein bound silent methylated OCT4 and NANOG promoters in fibroblasts, but not active demethylated promoters in ES cells. These data provide new evidence that mammalian AID is required for active DNA demethylation and initiation of nuclear reprogramming towards pluripotency in human somatic cells.


Improving hygiene in home deliveries in rural Ghana: how to build on current attitudes and practices.
Clean delivery of newborns is a key intervention for reducing infection-related neonatal mortality. Understanding local practices and beliefs is important for designing appropriate interventions. There are few data from Africa. This study explored delivery practices in Ghana to identify behaviors for intervention and to determine behavioral influencers. Data on the prevalence of clean delivery behaviors, collected through a demographic surveillance system, were analyzed for 2631 women who delivered at home within a 1-year period. Qualitative data on delivery practices were collected through birth narratives, in-depth interviews, and focus groups with recently delivered/pregnant women, traditional birth attendants, grandmothers, and husbands. Most women delivered on a covered surface (79%), had birth attendants who washed their hands (79%), cut the cord with a new blade (98%), and tied it with a new thread (90%). Eight percent of families practiced dry cord care. Families understood the importance of a clean delivery surface and many birth attendants knew the importance of hand-washing. Delivering on an uncovered surface was linked to impromptu deliveries and a belief that a swept floor is clean. Not washing hands was linked to rushing to help the woman, not being provided with soap, forgetfulness, and a belief among some that the babies are born dirty. The frequent application of products to the cord was nearly universal and respondents believed that applying nothing to the cord would have serious negative consequences. Delivery surfaces, hand-washing, and cord cutting and tying appear appropriate for the majority of women. Changing cord care practices is likely to be difficult unless replacement products are provided.


Characteristics of phase-change materials containing oxide nano-additives for thermal storage.
In this study, the authors report the production of nanocomposite-enhanced phase-change materials (NEPCMs) using the direct-synthesis method by mixing paraffin with alumina (Al2O3), titania (TiO2), silica (SiO2), and zinc oxide (ZnO) as the experimental samples. Al2O3, TiO2, SiO2, and ZnO were dispersed into three concentrations of 1.0, 2.0, and 3.0 wt.%. Through heat conduction and differential scanning calorimeter experiments to evaluate the effects of varying concentrations of the nano-additives on the heat conduction performance and thermal storage characteristics of NEPCMs, their feasibility for use in thermal storage was determined. The experimental results demonstrate that TiO2 is more effective than the other additives in enhancing both the heat conduction and thermal storage performance of paraffin for most of the experimental parameters. Furthermore, TiO2 reduces the melting onset temperature and increases the solidification onset temperature of paraffin. This allows the phase-change heat to be applicable to a wider temperature range, and the highest decreased ratio of phase-change heat is only 0.46%, compared to that of paraffin. Therefore, this study demonstrates that TiO2, added to paraffin to form NEPCMs, has significant potential for enhancing the thermal storage characteristics of paraffin.


Data mining on electronic health record databases for signal detection in pharmacovigilance: which events to monitor?
Data mining on electronic health records (EHRs) has emerged as a promising complementary method for post-marketing drug safety surveillance. The EU-ADR project, funded by the European Commission, is developing techniques that allow mining of EHRs for adverse drug events across different countries in Europe. Since mining on all possible events was considered to unduly increase the number of spurious signals, we wanted to create a ranked list of high-priority events. Scientific literature, medical textbooks, and websites of regulatory agencies were reviewed to create a preliminary list of events that are deemed important in pharmacovigilance. Two teams of pharmacovigilance experts independently rated each event on five criteria: 'trigger for drug withdrawal', 'trigger for black box warning', 'leading to emergency department visit or hospital admission', 'probability of event to be drug-related', and 'likelihood of death'. In case of disagreement, a consensus score was obtained. Ordinal scales between 0 and 3 were used for rating the criteria, and an overall score was computed to rank the events. An initial list comprising 23 adverse events was identified. After rating all the events and calculation of overall scores, a ranked list was established. The top-ranking events were: cutaneous bullous eruptions, acute renal failure, anaphylactic shock, acute myocardial infarction, and rhabdomyolysis. A ranked list of 23 adverse drug events judged as important in pharmacovigilance was created to permit focused data mining. The list will need to be updated periodically as knowledge on drug safety evolves and new issues in drug safety arise.


Interventions to increase testing, linkage to care and treatment of hepatitis C virus (HCV) infection among people in prisons: A systematic review.
While the burden of chronic hepatitis C virus (HCV) infection is significantly higher among people in prisons compared to the general population, testing and treatment uptake remain suboptimal. The aim of this systematic review was to synthesize evidence on the effectiveness of interventions to increase HCV testing, linkage to care and treatment uptake among people in prisons. We searched Medline (Ovid 1996-present), Embase (Ovid 1996-present), and the Cochrane Central Register of Controlled Trials for English language articles published between January 2007 and November 2017. Studies evaluating interventions to enhance HCV testing, linkage to care and treatment uptake for people in prison were included. Two independent reviewers evaluated articles selected for full-text review. Disagreements were resolved by consensus. A total of 475 unique articles were identified, 29 were eligible for full text review, and six studies were included. All but one study was conducted in the pre-direct-acting antiviral (DAA) era; no studies were conducted in low- or middle-income countries. Of the six studies, all but one focused on testing. Only two were randomised controlled trials; the remaining were single arm studies. Interventions to enhance HCV testing in prison settings included combination risk-based and birth-cohort screening strategies, on-site nurse-led opt-in screening clinics with pre-test counselling and education, and systematic dried blood spot testing. All interventions increased HCV testing, but risk of study bias was high in all studies. Interventions to enhance linkage to care included facilitated referral for HCV assessment and scheduling of specialist appointments; however, risk of study bias was critical. There is a lack of recent data on interventions to improve the HCV care cascade in people in prisons. With the introduction of short-course, well-tolerated DAAs, rigorous controlled studies evaluating interventions to improve testing, linkage and treatment uptake for people in prison are necessary.


Light availability prevails over soil fertility and structure in the performance of Asian knotweeds on riverbanks: new management perspectives.
Asian knotweeds (Fallopia spp.) are considered one of the world's most invasive species. Restoring habitats dominated by these exotic species requires a better understanding of the importance of abiotic factors controlling the invasive knotweeds performance. We used observational data obtained on the embankment of the Isère River (France) to study the performance of Fallopia spp. under different soil, light, and disturbance conditions. On the Isère riverbanks, light intensity assessed by light quantity transmitted through canopy was the most important factor explaining the variability observed on knotweed performance expressed as above-ground biomass per square meter. Asian knotweeds were more productive under intensive light conditions. Alternatively other factors such as mowing (twice a year), soil fertility, soil texture, position on the bank or exposure to the sun had no significant effect on knotweed biomass production. We conclude that decreasing light resources, for example, by increasing competitive pressure on sites dominated by Asian knotweeds could be included in management plans to control the populations of this invasive taxon.


Microbiome of the Black-Lipped Pearl Oyster Pinctada margaritifera, a Multi-Tissue Description With Functional Profiling.
Elucidating the role of prokaryotic symbionts in mediating host physiology has emerged as an important area of research. Since oysters are the world's most heavily cultivated bivalve molluscs, numerous studies have applied molecular techniques to understand the taxonomic and functional diversity of their associated bacteria. Here, we expand on this research by assessing the composition and putative functional profiles of prokaryotic communities from different organs/compartments of the black-lipped pearl oyster Pinctada margaritifera, a commercially important shellfish valued for cultured pearl production in the Pacific region. Seven tissues, in addition to mucous secretions, were targeted from P. margaritifera individuals: the gill, gonad, byssus gland, haemolymph, mantle, adductor muscle, mucus, and gut. Richness of bacterial Operational Taxonomic Units (OTUs) and phylogenetic diversity differed between host tissues, with mucous layers displaying the highest richness and diversity. This multi-tissues approach permitted the identification of consistent microbial members, together constituting the core microbiome of P. margaritifera, including Alpha- and Gammaproteobacteria, Flavobacteriia, and Spirochaetes. We also found a high representation of Endozoicimonaceae symbionts, indicating that they may be of particular importance to oyster health, survival and homeostasis, as in many other coral reef animals. Our study demonstrates that the microbial communities and their associated predicted functional profiles are tissue specific. Inferred physiological functions were supported by current physiological data available for the associated bacterial taxa specific to each tissue. This work provides the first baseline of microbial community composition in P. margaritifera, providing a solid foundation for future research into this commercially important species and emphasises the important effects of tissue differentiation in structuring the oyster microbiome.


Four crystal structures of human LLT1, a ligand of human NKR-P1, in varied glycosylation and oligomerization states.
Human LLT1 is a C-type lectin-like ligand of NKR-P1 (CD161, gene KLRB1), a C-type lectin-like receptor of natural killer cells. Using X-ray diffraction, the first experimental structures of human LLT1 were determined. Four structures of LLT1 under various conditions were determined: monomeric, dimeric deglycosylated after the first N-acetylglucosamine unit in two forms and hexameric with homogeneous GlcNAc2Man5 glycosylation. The dimeric form follows the classical dimerization mode of human CD69. The monomeric form keeps the same fold with the exception of the position of an outer part of the long loop region. The hexamer of glycosylated LLT1 consists of three classical dimers. The hexameric packing may indicate a possible mode of interaction of C-type lectin-like proteins in the glycosylated form.


Munchausen syndrome mimicking psychiatric disease with concomitant genuine physical illness.
Munchausen syndrome is a disorder in which patients intentionally produce symptoms mimicking physical or psychiatric illnesses with the aim to assume the sick role and to gain medical attention. Once a patient receives a Munchausen syndrome diagnosis every complaint made thence tends to be regarded with scepticism by clinical staff. However, it is possible that a bona fide illness, which might be disregarded, may coexist in these patients. We report a case of MS mimicking psychiatric disease with concomitant genuine acute physical illness. Despite the initial doubts about the veracity of the latter, due to its prompt recognition, treatment was successful.


Real-time teleteaching in medical physics.
Medical physics is a relatively small professional community, usually with a scarcity of expertise that could greatly benefit students entering the field. However, the reach of the profession can span great geographical distances, making the training of students a difficult task. In addition to the requirement of training new students, the evolving field of medical physics, with its many emerging advanced techniques and technologies, could benefit greatly from ongoing continuing education as well as consultation with experts.Many continuing education courses and workshops are constantly being offered, including many web-based study courses and virtual libraries. However, one mode of education and communication that has not been widely used is the real-time interactive process. Video-based conferencing systems do exist, but these usually require a substantial amount of effort and cost to set up.The authors have been working on promoting the ever-expanding capability of the Internet to facilitate the education of medical physics to students entering the field. A pilot project has been carried out for six years and reported previously. The project is a collaboration between the Department of Medical Physics at the Toronto Odette Cancer Centre in Canada and the Department of Biomedical Imaging at the University of Malaya in Malaysia. Since 2001, medical physics graduate students at the University of Malaya have been taught by lecturers from Toronto every year, using the Internet as the main tool of communication.The pilot study explored the different methods that can be used to provide real-time interactive remote education, and delivered traditional classroom lectures as well as hands-on workshops.Another similar project was started in 2007 to offer real-time teaching to a class of medical physics students at Wuhan University in Hubei, China. There are new challenges as well as new opportunities associated with this project. By building an inventory of tools and experiences, the intent is to broaden the real-time teleteaching method to serve a wide community so that future students entering the field can have efficient access to high-quality education that will benefit the profession in the long term.


The fusion protein SS18-SSX1 employs core Wnt pathway transcription factors to induce a partial Wnt signature in synovial sarcoma.
Expression of the SS18/SYT-SSX fusion protein is believed to underlie the pathogenesis of synovial sarcoma (SS). Recent evidence suggests that deregulation of the Wnt pathway may play an important role in SS but the mechanisms whereby SS18-SSX might affect Wnt signaling remain to be elucidated. Here, we show that SS18/SSX tightly regulates the elevated expression of the key Wnt target AXIN2 in primary SS. SS18-SSX is shown to interact with TCF/LEF, TLE and HDAC but not β-catenin in vivo and to induce Wnt target gene expression by forming a complex containing promoter-bound TCF/LEF and HDAC but lacking β-catenin. Our observations provide a tumor-specific mechanistic basis for Wnt target gene induction in SS that can occur in the absence of Wnt ligand stimulation.


Comparison of colistin monotherapy and non-colistin combinations in the treatment of multi-drug resistant Acinetobacter spp. bloodstream infections: a multicenter retrospective analysis.
To compare the efficacy of colistin (COL) monotherapy versus non-COL based combinations in the treatment of bloodstream infections (BSIs) due to multidrug resistant Acinetobacter spp.(MDR-A). Retrospective data of 107 MDR-A BSI cases from 27 tertiary centers in Turkey were included. 14-day mortality. Microbial eradication and clinical improvement. Thirty-six patients in the COL monotherapy (CM) group and 71 in the non-COL based combinations (NCC) group were included in the study. Mean age was 59.98 ± 20 years (range: 18-89) and 50.5% were male. Median duration of follow-up was 40 days (range: 9-297). The 14-day survival rates were 52.8% in CM and 47.23% in NCC group (P = 0.36). Microbiological eradication was achieved in 69% of CM and 83% of NCC group (P = 0.13). Treatment failure was detected in 22.9% of cases in both CM and NCC groups. Univariate analysis revealed that mean age (P = 0.001), Charlson comorbidity index (P = 0.03), duration of hospital stay before MDR-A BSI (P = 0.04), Pitt bacteremia score (P = 0.043) and Acute Physiology and Chronic Health Evaluation II score (P = 0.05) were significant in terms of 14-day mortality. Advanced age (P = 0.01) and duration of hospital stay before MDR-A BSI (P = 0.04) were independently associated with 14-day mortality in multivariate analysis. No significant difference was detected between CM and non-COL based combinations in the treatment of MDR-A BSIs in terms of efficacy and 14-day mortality.


Prospects for the diagnosis of breast cancer by noninvasive probing of calcifications using transmission Raman spectroscopy.
Breast calcifications can be found in both benign and malignant lesions, and the composition of these calcifications can indicate the possible disease state. As current practices such as mammography and histopathology examine the morphology of the specimen, they cannot reliably distinguish between the two types of calcification, which frequently are the only mammographic features that indicate the presence of a cancerous lesion. Raman spectroscopy is an optical technique capable of obtaining biochemical information of a sample in situ. We demonstrate for the first time the noninvasive recovery of Raman spectra of calcified materials buried within a chicken breast tissue slab 16 mm thick, achieved using transmission Raman spectroscopy. The spectra of both calcium hydroxyapatite (HAP) and calcium oxalate monohydrate (COM) are obtained and chemically identified. The experimental geometry and gross insensitivity of the Raman signal to the depth of the calcified lesion makes the concept potentially well suited for probing human female breasts, in conjunction with existing mammography or ultrasound, to provide complementary data in the early diagnosis of breast cancer.


[Analysis of blastomogenic activity of mammal carcinogens in Drosophila using the wts(P4) allele and RNA interference-induced p53 silencing].
The test for somatic mutagenesis and recombination in Drosophila is one of the widely used approaches for determination of possible carcinogenic effects of chemical compounds. The use of heterozygotes for mutant tumor suppressor gene wts enables more direct evaluation of the blastomogenic effects of chemical compounds, by tumor formation in the adult flies. This study presents evaluation of the SMART effectiveness upon the use of Drosophila heterozygotes for the wts(P4) gene, first included into the test system. The increase of the test resolution capacity compared to the literature data for the wts(P2) allele was observed. Using wts(P4) heterozygotes, a total of 20 carcinogenic compounds, and their slightly carcinogenic and noncarcinogenic analogs were tested. Specificity of the method was about 100%, and sensitivity depended on the type of the agent tested. The latter was absolute for the direct action carcinogens, with respect to carcinogens, requiring the metabolic activation. The sensitivity was elective and was limited by the presence of the enzymes capable of activating of these compounds. To increase the test sensitivity, the RNA interference-mediated silencing of the Drosophila p53 functional activity was successfully applied. Moreover, the frequency of wts tumor induction considerably increased both in spontaneous and induced mutagenesis conditions.


Active Warfighter Mental Health Lower in Mid-Career.
The present study investigated Special Operations Forces (SOF) combat Servicemember mental health at different SOF career stages in association with resilience. Fifty-eight SOF combat Service Members either entering SOF (career start; n=38) or multiple years with their SOF organization (mid-career; n=20) self-reported mild traumatic brain injury (TBI) history, resilience, subjective well-being, depression, anxiety, and posttraumatic stress. Poisson regression analyses were employed to test SOF career stage differences in each mental health symptom using resilience, while accounting for other pertinent military factors. There were significant interaction effects of SOF career stage and resilience on mental health symptoms. SOF career start combat Servicemembers endorsed lower depression and posttraumatic stress and higher subjective well-being with higher resilience, but these associations between resilience and mental health symptoms were not seen in SOF mid-career Servicemembers. Although preliminary, the adaptive association between resilience and mental health seemed to be blunted in combat Servicemembers having served multiple years in SOF. This information informs research to provide evaluation tools to support prophylactic performance and long-term health preservation in military populations.


Gamma Knife radiosurgery for posterior fossa meningiomas: a multicenter study.
Posterior fossa meningiomas represent a common yet challenging clinical entity. They are often associated with neurovascular structures and adjacent to the brainstem. Resection can be undertaken for posterior fossa meningiomas, but residual or recurrent tumor is frequent. Stereotactic radiosurgery (SRS) has been used to treat meningiomas, and this study evaluates the outcome of this approach for those located in the posterior fossa. At 7 medical centers participating in the North American Gamma Knife Consortium, 675 patients undergoing SRS for a posterior fossa meningioma were identified, and clinical and radiological data were obtained for these cases. Females outnumbered males at a ratio of 3.8 to 1, and the median patient age was 57.6 years (range 12-89 years). Prior resection was performed in 43.3% of the patient sample. The mean tumor volume was 6.5 cm(3), and a median margin dose of 13.6 Gy (range 8-40 Gy) was delivered to the tumor. At a mean follow-up of 60.1 months, tumor control was achieved in 91.2% of cases. Actuarial tumor control was 95%, 92%, and 81% at 3, 5, and 10 years after radiosurgery. Factors predictive of tumor progression included age greater than 65 years (hazard ratio [HR] 2.36, 95% CI 1.30-4.29, p = 0.005), prior history of radiotherapy (HR 5.19, 95% CI 1.69-15.94, p = 0.004), and increasing tumor volume (HR 1.05, 95% CI 1.01-1.08, p = 0.005). Clinical stability or improvement was achieved in 92.3% of patients. Increasing tumor volume (odds ratio [OR] 1.06, 95% CI 1.01-1.10, p = 0.009) and clival, petrous, or cerebellopontine angle location as compared with petroclival, tentorial, and foramen magnum location (OR 1.95, 95% CI 1.05-3.65, p = 0.036) were predictive of neurological decline after radiosurgery. After radiosurgery, ventriculoperitoneal shunt placement, resection, and radiation therapy were performed in 1.6%, 3.6%, and 1.5%, respectively. Stereotactic radiosurgery affords a high rate of tumor control and neurological preservation for patients with posterior fossa meningiomas. Those with a smaller tumor volume and no prior radiation therapy were more likely to have a favorable response after radiosurgery. Rarely, additional procedures may be required for hydrocephalus or tumor progression.


Vaccination program in the response to the 2007 equine influenza outbreak in Australia.
At the time of the initial notification of the occurrence of equine influenza (EI) in Australia in August 2007, vaccination was restricted to horses for which it was an import requirement and only with the approval of the state or territory Chief Veterinary Officer. This paper describes the complexities involved in the selection of a vaccine and its distribution. A combination of ring, predictive and blanket vaccination was implemented during the response. The specific vaccination programs, including its use in buffer zones and for movement of horses, the performance of the vaccine, any adverse reactions and the effect on exposure of vaccinated horses to circulating virus, are also described. Vaccination is considered to have made a valuable contribution to the containment and subsequent eradication of EI from Australia and to risk management in the resumption of horse activities in affected areas from December 2007. Movement restrictions and other biosecurity measures were critically important in controlling the spread of EI and contributing to its eventual eradication, and vaccination was an aid to these measures.


Discovery of a Glucocorticoid Receptor (GR) Activity Signature Using Selective GR Antagonism in ER-Negative Breast Cancer.
Purpose: Although high glucocorticoid receptor (GR) expression in early-stage estrogen receptor (ER)-negative breast cancer is associated with shortened relapse-free survival (RFS), how associated GR transcriptional activity contributes to aggressive breast cancer behavior is not well understood. Using potent GR antagonists and primary tumor gene expression data, we sought to identify a tumor-relevant gene signature based on GR activity that would be more predictive than GR expression alone.Experimental Design: Global gene expression and GR ChIP-sequencing were performed to identify GR-regulated genes inhibited by two chemically distinct GR antagonists, mifepristone and CORT108297. Differentially expressed genes from MDA-MB-231 cells were cross-evaluated with significantly expressed genes in GR-high versus GR-low ER-negative primary breast cancers. The resulting subset of GR-targeted genes was analyzed in two independent ER-negative breast cancer cohorts to derive and then validate the GR activity signature (GRsig).Results: Gene expression pathway analysis of glucocorticoid-regulated genes (inhibited by GR antagonism) revealed cell survival and invasion functions. GR ChIP-seq analysis demonstrated that GR antagonists decreased GR chromatin association for a subset of genes. A GRsig that comprised n = 74 GR activation-associated genes (also reversed by GR antagonists) was derived from an adjuvant chemotherapy-treated Discovery cohort and found to predict probability of relapse in a separate Validation cohort (HR = 1.9; P = 0.012).Conclusions: The GRsig discovered herein identifies high-risk ER-negative/GR-positive breast cancers most likely to relapse despite administration of adjuvant chemotherapy. Because GR antagonism can reverse expression of these genes, we propose that addition of a GR antagonist to chemotherapy may improve outcome for these high-risk patients. Clin Cancer Res; 24(14); 3433-46. ©2018 AACR.


Rare paraneoplastic erythroderma associated with ectopic neuron-specific enolase deposition in basal cells.
The cutaneous symptom of the paraneoplastic erythroderma can be the only symptom of a malignancy. Although many cases associated with malignancies have been reported, the pathogenesis of cancer related erythroderma is still unclear. Herein we presented a patient with large cell neuroendocrine carcinoma (LCNEC) of the lung and contemporary severe erythroderma. The patient suffered from skin erythema and scaling all over the body and the cutaneous lesions recovered completely after 3 weeks of surgery. Strong expression of neuron-specific enolase (NSE, 2+ positive) was found in both primary cancer and basal cells of the preoperative skin. Three months later, postoperative skin biopsy presented nearly normal skin tissues, accompanied with a negative expression of NSE. Nine months after surgery, cancer recurred in the liver and brain with the first symptom of skin erythema and scaling. The pathology of liver biopsy tissues illustrated the LCNEC and 3+ positive expression of NSE. The skin biopsy tissues showed 2+ positive stain of NSE. Evaluation after two cycles of chemotherapy showed marked improvement in erythroderma and reduction of tumor volume. However, the patient experienced recurrent worsening of erythroderma when chemotherapy was terminated due to severe myelosuppression. Eleven months after surgery, the patient died of cancer cachexia and multiple organ failure. To our knowledge, this was the first case of paraneoplastic erythroderma associated with LCNEC of lung. Furthermore, we firstly discovered that the deposition of NSE in basal cells might be a crucial pathogenic factor of erythroderma.


Dynamic palmitoylation events following T-cell receptor signaling.
Palmitoylation is the reversible addition of palmitate to cysteine via a thioester linkage. The reversible nature of this modification makes it a prime candidate as a mechanism for regulating signal transduction in T-cell receptor signaling. Following stimulation of the T-cell receptor we find a number of proteins are newly palmitoylated, including those involved in vesicle-mediated transport and Ras signal transduction. Among these stimulation-dependent palmitoylation targets are the v-SNARE VAMP7, important for docking of vesicular LAT during TCR signaling, and the largely undescribed palmitoyl acyltransferase DHHC18 that is expressed in two isoforms in T cells. Using our newly developed On-Plate Palmitoylation Assay (OPPA), we show DHHC18 is capable of palmitoylating VAMP7 at Cys183. Cellular imaging shows that the palmitoylation-deficient protein fails to be retained at the Golgi and to localize to the immune synapse upon T cell activation.


Comparison of the bacterial flora of the duodenum in healthy cats and cats with signs of gastrointestinal tract disease.
To determine whether a colony environment predisposes healthy cats to high bacterial counts, including counts of obligate anaerobes, in the duodenum and whether increased numbers of bacteria could be found in the duodenum of cats with signs of chronic gastrointestinal tract disease. Prospective study. 20 healthy control cats (10 from a colony environment and 10 pet cats) and 19 cats with a history of chronic gastrointestinal tract disease. Undiluted duodenal fluid was quantitatively and qualitatively assessed by bacteriologic culture under aerobic and anaerobic conditions. Serum concentrations of cobalamin and folate were also measured. Significant differences were not detected in the numbers of bacteria found in the duodenum of cats housed in a colony environment, compared with pet cats fed an identical diet prior to sampling. All healthy cats were, therefore, combined into 1 control group. Compared with healthy cats, cats with clinical signs of gastrointestinal tract disease had significantly lower counts of microaerophilic bacteria, whereas total, anaerobic, and aerobic bacterial counts were not significantly different. None of the cats with disease had total bacterial counts higher than expected from the range established in the control cats. Differences were not detected in regard to serum folate or cobalamin concentrations between diseased and healthy cats. These findings indicated that healthy colony cats and pet cats have high numbers of bacteria in the duodenum, including high numbers of obligate anaerobes. Our findings also suggest that bacterial overgrowth in the small intestine is not a common clinical syndrome in cats with chronic nonobstructive gastrointestinal tract disease.