Efficacy and safety of ticagrelor for long-term secondary prevention of atherothrombotic events in relation to renal function: insights from the PEGASUS-TIMI 54 trial.
We evaluated the relationship of renal function and ischaemic and bleeding risk as well as the efficacy and safety of ticagrelor in stable patients with prior myocardial infarction (MI). Patients with a history of MI 1-3 years prior from PEGASUS-TIMI 54 were stratified based on estimated glomerular filtration rate (eGFR), with <60 mL/min/1.73 m(2) pre-specified for analysis of the effect of ticagrelor on the primary efficacy composite of cardiovascular death, MI, or stroke (major adverse cardiovascular events, MACE) and the primary safety endpoint of TIMI major bleeding. Of 20 898 patients, those with eGFR <60 (N = 4849, 23.2%) had a greater risk of MACE at 3 years relative to those without, which remained significant after multivariable adjustment (hazard ratio, HRadj 1.54, 95% confidence interval, CI 1.27-1.85, P < 0.001). The relative risk reduction in MACE with ticagrelor was similar in those with eGFR <60 (ticagrelor pooled vs. placebo: HR 0.81; 95% CI 0.68-0.96) vs. ≥60 (HR 0.88; 95% CI 0.77-1.00, Pinteraction = 0.44). However, due to the greater absolute risk in the former group, the absolute risk reduction with ticagrelor was higher: 2.7 vs. 0.63%. Bleeding tended to occur more frequently in patients with renal dysfunction. The absolute increase in TIMI major bleeding with ticagrelor was similar in those with and without eGFR <60 (1.19 vs. 1.43%), whereas the excess of minor bleeding tended to be more pronounced (1.93 vs. 0.69%). In patients with a history of MI, patients with renal dysfunction are at increased risk of MACE and consequently experience a particularly robust absolute risk reduction with long-term treatment with ticagrelor.


Embryonic DNA methylation: insights from the genomics era.
DNA methylation is an epigenetic mark associated with transcriptional repression and essential for proper vertebrate development. Because of its association with human development and disease, DNA methylation has stayed in the research focus for almost half a century. However, it is only since very recently that we are able to generate genome-wide maps of this mark and gain insight into the many roles DNA methylation plays during the vertebrate life cycle. The recent adaptation of next-generation sequencing technologies to global DNA methylation profiling has spurred a flurry of studies conducted on various model systems. The recent publications of zebrafish base-pair resolution methylomes provided key advances in our understanding of how DNA methylation is remodeled during early vertebrate embryogenesis and highlighted important differences in developmental strategies between mammals and lower vertebrates. Here, we review our understanding of the role of DNA methylation during vertebrate development and discuss the insights obtained from almost a decade of methylome studies.


Rapid epidemic expansion of the SARS-CoV-2 Omicron variant in southern Africa.
The SARS-CoV-2 epidemic in southern Africa has been characterized by three distinct waves. The first was associated with a mix of SARS-CoV-2 lineages, while the second and third waves were driven by the Beta (B.1.351) and Delta (B.1.617.2) variants, respectively1-3. In November 2021, genomic surveillance teams in South Africa and Botswana detected a new SARS-CoV-2 variant associated with a rapid resurgence of infections in Gauteng province, South Africa. Within three days of the first genome being uploaded, it was designated a variant of concern (Omicron, B.1.1.529) by the World Health Organization and, within three weeks, had been identified in 87 countries. The Omicron variant is exceptional for carrying over 30 mutations in the spike glycoprotein, which are predicted to influence antibody neutralization and spike function4. Here we describe the genomic profile and early transmission dynamics of Omicron, highlighting the rapid spread in regions with high levels of population immunity.


Do ants need to estimate the geometrical properties of trail bifurcations to find an efficient route? A swarm robotics test bed.
Interactions between individuals and the structure of their environment play a crucial role in shaping self-organized collective behaviors. Recent studies have shown that ants crossing asymmetrical bifurcations in a network of galleries tend to follow the branch that deviates the least from their incoming direction. At the collective level, the combination of this tendency and the pheromone-based recruitment results in a greater likelihood of selecting the shortest path between the colony's nest and a food source in a network containing asymmetrical bifurcations. It was not clear however what the origin of this behavioral bias is. Here we propose that it results from a simple interaction between the behavior of the ants and the geometry of the network, and that it does not require the ability to measure the angle of the bifurcation. We tested this hypothesis using groups of ant-like robots whose perceptual and cognitive abilities can be fully specified. We programmed them only to lay down and follow light trails, avoid obstacles and move according to a correlated random walk, but not to use more sophisticated orientation methods. We recorded the behavior of the robots in networks of galleries presenting either only symmetrical bifurcations or a combination of symmetrical and asymmetrical bifurcations. Individual robots displayed the same pattern of branch choice as individual ants when crossing a bifurcation, suggesting that ants do not actually measure the geometry of the bifurcations when travelling along a pheromone trail. Finally at the collective level, the group of robots was more likely to select one of the possible shorter paths between two designated areas when moving in an asymmetrical network, as observed in ants. This study reveals the importance of the shape of trail networks for foraging in ants and emphasizes the underestimated role of the geometrical properties of transportation networks in general.


Enisamium is an inhibitor of the SARS-CoV-2 RNA polymerase and shows improvement of recovery in COVID-19 patients in an interim analysis of a clinical trial.
Pandemic SARS-CoV-2 causes a mild to severe respiratory disease called Coronavirus Disease 2019 (COVID-19). Control of SARS-CoV-2 spread will depend on vaccine-induced or naturally acquired protective herd immunity. Until then, antiviral strategies are needed to manage COVID-19, but approved antiviral treatments, such as remdesivir, can only be delivered intravenously. Enisamium (laboratory code FAV00A, trade name Amizon®) is an orally active inhibitor of influenza A and B viruses in cell culture and clinically approved in countries of the Commonwealth of Independent States. Here we show that enisamium can inhibit SARS-CoV-2 infections in NHBE and Caco-2 cells. In vitro, the previously identified enisamium metabolite VR17-04 directly inhibits the activity of the SARS-CoV-2 RNA polymerase. Docking and molecular dynamics simulations suggest that VR17-04 prevents GTP and UTP incorporation. To confirm enisamium's antiviral properties, we conducted a double-blind, randomized, placebo-controlled trial in adult, hospitalized COVID-19 patients, which needed medical care either with or without supplementary oxygen. Patients received either enisamium (500 mg per dose) or placebo for 7 days. A pre-planned interim analysis showed in the subgroup of patients needing supplementary oxygen (n = 77) in the enisamium group a mean recovery time of 11.1 days, compared to 13.9 days for the placebo group (log-rank test; p=0.0259). No significant difference was found for all patients (n = 373) or those only needing medical care (n = 296). These results thus suggest that enisamium is an inhibitor of SARS-CoV-2 RNA synthesis and that enisamium treatment shortens the time to recovery for COVID-19 patients needing oxygen.


Toxicological Profiling of Highly Purified Metallic and Semiconducting Single-Walled Carbon Nanotubes in the Rodent Lung and E. coli.
The electronic properties of single-walled carbon nanotubes (SWCNTs) are potentially useful for electronics, optics, and sensing applications. Depending on the chirality and diameter, individual SWCNTs can be classified as semiconducting (S-SWCNT) or metallic (M-SWCNT). From a biological perspective, the hazard profiling of purified metallic versus semiconducting SWCNTs has been pursued only in bacteria, with the conclusion that aggregated M-SWCNTs are more damaging to bacterial membranes than S-SWCNTs. However, no comparative studies have been performed in a mammalian system, where most toxicity studies have been undertaken using relatively crude SWCNTs that include a M:S mix at 1:2 ratio. In order to compare the toxicological impact of SWCNTs sorted to enrich them for each of the chirality on pulmonary cells and the intact lung, we used density gradient ultracentrifugation and extensive rinsing to prepare S- and M-SWCNTs that are >98% purified. In vitro screening showed that both tube variants trigger similar amounts of interleukin 1β (IL-1β) and transforming growth factor (TGF-β1) production in THP-1 and BEAS-2B cells, without cytotoxicity. Oropharyngeal aspiration confirmed that both SWCNT variants induce comparable fibrotic effects in the lung and abundance of IL-1β and TGF-β1 release in the bronchoalveolar lavage fluid. There was also no change in the morphology, membrane integrity, and viability of E. coli, in contradistinction to the previously published effects of aggregated tubes on the bacterial membrane. Collectively, these data indicate that the electronic properties and chirality do not independently impact SWCNT toxicological impact in the lung, which is of significance to the safety assessment and incremental use of purified tubes by industry.


Neural correlates of real-world route learning.
Classical theories of spatial microgenesis (Siegel and White, 1975) posit that information about landmarks and the paths between them is acquired prior to the establishment of more holistic survey-level representations. To test this idea, we examined the neural and behavioral correlates of landmark and path encoding during a real-world route learning episode. Subjects were taught a novel 3 km route around the University of Pennsylvania campus and then brought to the laboratory where they performed a recognition task that required them to discriminate between on-route and off-route buildings. Each building was preceded by a masked prime, which could either be the building that immediately preceded the target building along the route or immediately succeeded it. Consistent with previous reports using a similar paradigm in a virtual environment (Janzen and Weststeijn, 2007), buildings at navigational decision points (DPs) were more easily recognized than non-DP buildings and recognition was facilitated by in-route vs. against-route primes. Functional magnetic resonance imaging (fMRI) data collected during the recognition task revealed two effects of interest: first, greater response to DP vs. non-DP buildings in a wide network of brain regions previously implicated in spatial processing; second, a significant interaction between building location (DP vs. non-DP) and route direction (in-route vs. against-route) in a retrosplenial/parietal-occipital sulcus region previously labeled the retrosplenial complex (RSC). These results indicate that newly learned real-world routes are coded in terms of paths between decision points and suggest that the RSC may be a critical locus for integrating landmark and path information.


Comparative genomic analysis of the thermophilic biomass-degrading fungi Myceliophthora thermophila and Thielavia terrestris.
Thermostable enzymes and thermophilic cell factories may afford economic advantages in the production of many chemicals and biomass-based fuels. Here we describe and compare the genomes of two thermophilic fungi, Myceliophthora thermophila and Thielavia terrestris. To our knowledge, these genomes are the first described for thermophilic eukaryotes and the first complete telomere-to-telomere genomes for filamentous fungi. Genome analyses and experimental data suggest that both thermophiles are capable of hydrolyzing all major polysaccharides found in biomass. Examination of transcriptome data and secreted proteins suggests that the two fungi use shared approaches in the hydrolysis of cellulose and xylan but distinct mechanisms in pectin degradation. Characterization of the biomass-hydrolyzing activity of recombinant enzymes suggests that these organisms are highly efficient in biomass decomposition at both moderate and high temperatures. Furthermore, we present evidence suggesting that aside from representing a potential reservoir of thermostable enzymes, thermophilic fungi are amenable to manipulation using classical and molecular genetics.


Baby Business: a randomised controlled trial of a universal parenting program that aims to prevent early infant sleep and cry problems and associated parental depression.
Infant crying and sleep problems (e.g. frequent night waking, difficulties settling to sleep) each affect up to 30% of infants and often co-exist. They are costly to manage and associated with adverse outcomes including postnatal depression symptoms, early weaning from breast milk, and later child behaviour problems. Preventing such problems could improve these adverse outcomes and reduce costs to families and the health care system. Anticipatory guidance-i.e. providing parents with information about normal infant sleep and cry patterns, ways to encourage self-settling in infants, and ways to develop feeding and settling routines before the onset of problems-could prevent such problems. This paper outlines the protocol for our study which aims to test an anticipatory guidance approach. 750 families from four Local Government Areas in Melbourne, Australia have been randomised to receive the Baby Business program (intervention group) or usual care (control group) offered by health services. The Baby Business program provides parents with information about infant sleep and crying via a DVD and booklet (mailed soon after birth), telephone consultation (at infant age 6-8 weeks) and parent group session (at infant age 12 weeks). All English speaking parents of healthy newborn infants born at > 32 weeks gestation and referred by their maternal and child health nurse at their first post partum home visit (day 7-10 postpartum), are eligible. The primary outcome is parent report of infant night time sleep as a problem at four months of age and secondary outcomes include parent report of infant daytime sleep or crying as a problem, mean duration of infant sleep and crying/24 hours, parental depression symptoms, parent sleep quality and quantity and health service use. Data will be collected at two weeks (baseline), four months and six months of age. An economic evaluation using a cost-consequences approach will, from a societal perspective, compare costs and health outcomes between the intervention and control groups. To our knowledge this is the first randomised controlled trial of a program which aims to prevent both infant sleeping and crying problems and associated postnatal depression symptoms. If effective, it could offer an important public health prevention approach to these common, distressing problems.


Pharmacological interventions for primary biliary cholangitis: an attempted network meta-analysis.
Primary biliary cholangitis (previously primary biliary cirrhosis) is a chronic liver disease caused by the destruction of small intra-hepatic bile ducts resulting in stasis of bile (cholestasis), liver fibrosis, and liver cirrhosis. The optimal pharmacological treatment of primary biliary cholangitis remains uncertain. To assess the comparative benefits and harms of different pharmacological interventions in the treatment of primary biliary cholangitis through a network meta-analysis and to generate rankings of the available pharmacological interventions according to their safety and efficacy. However, it was not possible to assess whether the potential effect modifiers were similar across different comparisons. Therefore, we did not perform the network meta-analysis, and instead, assessed the comparative benefits and harms of different interventions using standard Cochrane methodology. We searched the Cochrane Central Register of Controlled Trials (CENTRAL; 2017, Issue 2), MEDLINE, Embase, Science Citation Index Expanded, World Health Organization International Clinical Trials Registry Platform, and randomised controlled trials registers to February 2017 to identify randomised clinical trials on pharmacological interventions for primary biliary cholangitis. We included only randomised clinical trials (irrespective of language, blinding, or publication status) in participants with primary biliary cholangitis. We excluded trials which included participants who had previously undergone liver transplantation. We considered any of the various pharmacological interventions compared with each other or with placebo or no intervention. We used standard methodological procedures expected by Cochrane. We calculated the odds ratio (OR) and rate ratio with 95% confidence intervals (CI) using both fixed-effect and random-effects models based on available-participant analysis with Review Manager 5. We assessed risk of bias according to Cochrane, controlled risk of random errors with Trial Sequential Analysis, and assessed the quality of the evidence using GRADE. We identified 74 trials including 5902 participants that met the inclusion criteria of this review. A total of 46 trials (4274 participants) provided information for one or more outcomes. All the trials were at high risk of bias in one or more domains. Overall, all the evidence was low or very low quality. The proportion of participants with symptoms varied from 19.9% to 100% in the trials that reported this information. The proportion of participants who were antimitochondrial antibody (AMA) positive ranged from 80.8% to 100% in the trials that reported this information. It appeared that most trials included participants who had not received previous treatments or included participants regardless of the previous treatments received. The follow-up in the trials ranged from 1 to 96 months.The proportion of people with mortality (maximal follow-up) was higher in the methotrexate group versus the no intervention group (OR 8.83, 95% CI 1.01 to 76.96; 60 participants; 1 trial; low quality evidence). The proportion of people with mortality (maximal follow-up) was lower in the azathioprine group versus the no intervention group (OR 0.56, 95% CI 0.32 to 0.98; 224 participants; 2 trials; I2 = 0%; low quality evidence). However, it has to be noted that a large proportion of participants (25%) was excluded from the trial that contributed most participants to this analysis and the results were not reliable. There was no evidence of a difference in any of the remaining comparisons. The proportion of people with serious adverse events was higher in the D-penicillamine versus no intervention group (OR 28.77, 95% CI 1.57 to 526.67; 52 participants; 1 trial; low quality evidence). The proportion of people with serious adverse events was higher in the obeticholic acid plus ursodeoxycholic acid (UDCA) group versus the UDCA group (OR 3.58, 95% CI 1.02 to 12.51; 216 participants; 1 trial; low quality evidence). There was no evidence of a difference in any of the remaining comparisons for serious adverse events (proportion) or serious adverse events (number of events). None of the trials reported health-related quality of life at any time point. nine trials had no special funding or were funded by hospital or charities; 31 trials were funded by pharmaceutical companies; and 34 trials provided no information on source of funding. Based on very low quality evidence, there is currently no evidence that any intervention is beneficial for primary biliary cholangitis. However, the follow-up periods in the trials were short and there is significant uncertainty in this issue. Further well-designed randomised clinical trials are necessary. Future randomised clinical trials ought to be adequately powered; performed in people who are generally seen in the clinic rather than in highly selected participants; employ blinding; avoid post-randomisation dropouts or planned cross-overs; should have sufficient follow-up period (e.g. five or 10 years or more); and use clinically important outcomes such as mortality, health-related quality of life, cirrhosis, decompensated cirrhosis, and liver transplantation. Alternatively, very large groups of participants should be randomised to facilitate shorter trial duration.


FRET-based calcium imaging: a tool for high-throughput/content phenotypic drug screening in Alzheimer disease.
Perturbed intracellular store calcium homeostasis is suggested to play a major role in the pathophysiology of Alzheimer disease (AD). A number of mechanisms have been suggested to underlie the impairment of endoplasmic reticulum calcium homeostasis associated with familial AD-linked presenilin 1 mutations (FAD-PS1). Without aiming at specifically targeting any of those pathophysiological mechanisms in particular, we rather performed a high-throughput phenotypic screen to identify compounds that can reverse the exaggerated agonist-evoked endoplasmic reticulum calcium release phenotype in HEK293 cells expressing FAD-PS1. For that purpose, we developed a fully automated high-throughput calcium imaging assay using a fluorescence resonance energy transfer-based calcium indicator at single-cell resolution. This novel robust assay offers a number of advantages compared with the conventional calcium measurement screening technologies. The assay was employed in a large-scale screen with a library of diverse compounds comprising 20,000 low-molecular-weight molecules, which resulted in the identification of 52 primary hits and 4 lead structures. In a secondary assay, several hits were found to alter the amyloid β (Aβ) production. In view of the recent failure of AD drug candidates identified by target-based approaches, such a phenotypic drug discovery paradigm may present an attractive alternative for the identification of novel AD therapeutics.


Unicompartmental knee arthroplasties implanted for osteoarthritis with partial loss of joint space have high re-operation rates.
The indications and contraindications for unicompartmental knee arthroplasty (UKA) are controversial. The aim of the study was to determine the risk factors for re-operation in our practice. A series of 113 medial UKAs with mean follow-up of 63 months were reviewed retrospectively. Pre-operatively all knees had radiographic or arthroscopic evidence of severe cartilage damage. The re-operation rate was not related to age, gender, arthroscopic finding or body mass index. It was related to the joint space on pre-operative standing weight bearing radiographs taken in extension. The re-operation rate was 6 (95% CI 2.1-17, P<0.001) times higher when the thickness of the pre-operative medial joint space was >2 mm rather than ≤2 mm. It was 8 (95% CI 2.8-22.5, P<0.001) times higher when the thickness of the pre-operative medial space was >40% of the thickness of the lateral space. The ratio of pre-operative joint spaces has a greater influence on revision rate than the absolute measurement and is independent of radiographic magnification or the patient's normal cartilage thickness. We therefore recommend that, in medial knee osteoarthritis, UKA should only be used if the pre-operative medial joint space on standing radiographs is ≤40% of the lateral joint space, even if severe cartilage damage is seen arthroscopically.


Time-Cumulative Toxicity of Neonicotinoids: Experimental Evidence and Implications for Environmental Risk Assessments.
Our mechanistic understanding of the toxicity of chemicals that target biochemical and/or physiological pathways, such as pesticides and medical drugs is that they do so by binding to specific molecules. The nature of the latter molecules (e.g., enzymes, receptors, DNA, proteins, etc.) and the strength of the binding to such chemicals elicit a toxic effect in organisms, which magnitude depends on the doses exposed in a given timeframe. While dose and time of exposure are critical factors determining the toxicity of pesticides, different types of chemicals behave differently. Experimental evidence demonstrates that the toxicity of neonicotinoids increases with exposure time as much as with the dose, and therefore it has been described as time-cumulative toxicity. Examples for aquatic and terrestrial organisms are shown here. This pattern of toxicity, also found among carcinogenic compounds and other toxicants, has been ignored in ecotoxicology and risk assessments for a long time. The implications of the time-cumulative toxicity of neonicotinoids on non-target organisms of aquatic and terrestrial environments are far reaching. Firstly, neonicotinoids are incompatible with integrated pest management (IPM) approaches and secondly regulatory assessments for this class of compounds cannot be based solely on exposure doses but need also to take into consideration the time factor.


[Professional opinion about hospitalising Latin-American immigrant children in Andalucía, Spain].
There has been an increase in paediatric hospital attention being provided for Latin-American immigrant patients in Spain during the recent years. This work was aimed at ascertaining professionals' opinion regarding this population's specific and differential needs concerning Latin-American immigrant children being hospitalised. The study consisted of a qualitative, phenomenological type of investigation, based on semi-structured interviews of key informers in Andalusia (Spain). The subjects of the study were professionals from paediatric services in hospital centres in Andalusia. One of the most prominent (positive) results revealed that the Latin-American immigrant population presented less difficulties regarding hospital functioning when compared to other immigrant groups. The psychological aspects which had most impact on paediatric hospitalisation would be emotional alterations, such as the presence of anxiety and stress, or other aspects, like a lack of trust when being hospitalised.


State funding for health information technology and selected ambulatory healthcare quality measures.
Previous studies on the effects of health information technology (health IT) on ambulatory quality have had mixed results. New York State has invested heavily in health IT throughout the State, creating a unique opportunity to assess effects on health care quality across multiple communities. To determine any association between primary care providers' receipt of funding from New York State's Healthcare Efficiency and Affordability Law for New Yorkers Program (HEAL NY) and ambulatory quality of care. A statewide, longitudinal cohort study of primary care physicians in New York State was conducted. Data regarding which primary care physicians received funding through the HEAL NY program (Phase 5 or Phase 10) in 2008 or 2009 were obtained from the New York State Department of Health. Health care quality in 2010 was measured using claims data that had been aggregated across 7 commercial health plans across the state. Physicians were divided into 2 groups, based on receipt of HEAL funding (yes/no). Any association was measured between study group and each of 7 quality measures, all of which appear in the Stage 1 federal Meaningful Use program. Negative binomial regression was used, adjusting for provider gender and specialty. The study included 3,988 primary care providers, of whom 863 (22%) had received HEAL NY funding. The HEAL-funded physicians provided higher quality of care on 5 of the 7 measures: breast cancer screening, eye exams in patients with diabetes, nephropathy screening in patients with diabetes, influenza vaccination and pneumococcal vaccination (p<0.0001 for all adjusted comparisons). The HEAL-funded group provided higher quality of care by an absolute 2 to 6 percentage points per measure for those 5 measures. Primary care physicians who received state funding for health IT provided higher quality of care than those who did not receive such funding.


Visual imagery in hemianopic patients.
In this article we report some findings about visual imagery in patients with stable homonymous hemianopia compared to healthy control subjects. These findings were obtained by analyzing the gaze control through recording of eye movements in different phases of viewing and imagery. We used six different visual stimuli for the consecutive viewing and imagery phases. With infrared oculography, we recorded eye movements during this presentation phase and in three subsequent imagery phases in absence of the stimulus. Analyzing the basic parameters of the gaze sequences (known as "scanpaths"), we discovered distinct characteristics of the "viewing scanpaths" and the "imagery scanpaths" in both groups, which suggests a reduced extent of the image within the cognitive representation. We applied different similarity measures (string/vector string editing, Markov analysis). We found a "progressive consistency of imagery," shown through raising similarity values for the comparison of the late imagery scanpaths. This result suggests a strong top-down component in picture exploration: In both groups, healthy subjects and hemianopic patients, a mental model of the viewed picture must evolve very soon and substantially determine the eye movements. As our hemianopic patients showed analogous results to the normal subjects, we conclude that these patients are well adjusted to their deficit and, despite their perceptual defect, have a preserved cognitive representation, which follows the same top-down vision strategies in the process of visual imagery.


The mild stress of chronic prenatal injections may have additive effects on drugs administered during pregnancy to alter brain sexual differentiation.
In order to clarify the effect of the prenatal (PN) treatment of the drug 1,4,6-androstatriene-3,17-dione (ATD) which blocks the conversion of testosterone into estradiol on male sexual behavior of the rats offsprings, from the effect of the mild stress induced by the PN administration of the Propylene glycol (PG), the vehicle used to dissolve ATD. Pregnant Wistar rats were divided into three groups. The CON group did not receive any kind of treatment. The other two groups (PG and ATD) were injected i.p. during gestation (days 11-22) with 0 and 5 mg of ATD, dissolved in 0.1 ml of PG, respectively, doses reported by other authors. Sexual performance of the male pups was analyzed three months later in four successive tests. In the first sexual test of these naive rats, the percentage of males mounting, intromitting, ejaculating and the ejaculation frequency of the ATD group decreased significantly in comparison with the CON group. Also in the first and 4th tests, mounting, intromission and ejaculation latencies, as in the post-ejaculatory refractory period, ATD group, was significantly longer in comparison with the CON group. PG males showed a male sexual behavior (MSB) similar to that observed in the ATD group, but the differences did not reach statistical significance when they were compared with the CON group. We considered that the PN stress induced by the daily administration of PG and ATD, results in a slower execution of the MSB in both groups and avoid distinguish the effect of the ATD. Then chronic PN injections, as a route of administration, could act as mild stressor and may have additive effects on drugs affecting brain sexual differentiation.


Major life-history transitions by deterministic directional natural selection.
For large-scale evolution on Earth there has been a directional change where simple self-replicators evolved into large organisms with high metabolic rates and long pre-reproductive periods. Associated with this increase there have been major life-history transitions from asomatic, non-senescing, haploid, and asexually reproducing organisms to somatic, senescing, diploid, and sexually reproducing organisms with male and female individuals. Using a game theoretical model, it is shown that this trajectory can be explained by deterministic natural selection as it arises from first principles of the self-replication process in mobile organisms. It is shown (i) that selection for an increase in the energetic state of the organism puts a direction to evolution, (ii) that selection by density-dependent competitive interactions can explain the major life-history transitions as a function of the energetic state of the organism, and (iii) that the two selection processes combined can explain an exponentially increasing body mass. It is also shown (iv) why, for the case with an increasing body mass, we may expect many life histories to evolve in accordance with the exponents of the body mass allometries, (v) that an upper constraint on the body mass and metabolic rate can induce an additional transition into eusocial communities, and (vi) that the evolutionary trajectory is likely reversible with backward evolution during periods of environmental crises.


The nuclear effector of Wnt-signaling, Tcf1, functions as a T-cell-specific tumor suppressor for development of lymphomas.
The HMG-box factor Tcf1 is required during T-cell development in the thymus and mediates the nuclear response to Wnt signals. Tcf1(-/-) mice have previously been characterized and show developmental blocks at the CD4-CD8- double negative (DN) to CD4+CD8+ double positive transition. Due to the blocks in T-cell development, Tcf1(-/-) mice normally have a very small thymus. Unexpectedly, a large proportion of Tcf1(-/-) mice spontaneously develop thymic lymphomas with 50% of mice developing a thymic lymphoma/leukemia at the age of 16 wk. These lymphomas are clonal, highly metastatic, and paradoxically show high Wnt signaling when crossed with Wnt reporter mice and have high expression of Wnt target genes Lef1 and Axin2. In wild-type thymocytes, Tcf1 is higher expressed than Lef1, with a predominance of Wnt inhibitory isoforms. Loss of Tcf1 as repressor of Lef1 leads to high Wnt activity and is the initiating event in lymphoma development, which is exacerbated by activating Notch1 mutations. Thus, Notch1 and loss of Tcf1 functionally act as collaborating oncogenic events. Tcf1 deficiency predisposes to the development of thymic lymphomas by ectopic up-regulation of Lef1 due to lack of Tcf1 repressive isoforms and frequently by cooperating activating mutations in Notch1. Tcf1 therefore functions as a T-cell-specific tumor suppressor gene, besides its established role as a Wnt responsive transcription factor. Thus, Tcf1 acts as a molecular switch between proliferative and repressive signals during T-lymphocyte development in the thymus.


Working memory in children: tracing age differences and special educational needs to parameters of a formal model.
Parameters of a formal working-memory model were estimated for verbal and spatial memory updating of children. The model proposes interference though feature overwriting and through confusion of whole elements as the primary cause of working-memory capacity limits. We tested 2 age groups each containing 1 group of normal intelligence and 1 deficit group. For young children the deficit was developmental dyslexia; for older children it was a general learning difficulty. The interference model predicts less interference through overwriting but more through confusion of whole elements for the dyslexic children than for their age-matched controls. Older children exhibited less interference through confusion of whole elements and a higher processing rate than young children, but general learning difficulty was associated with slower processing than in the age-matched control group. Furthermore, the difference between verbal and spatial updating mapped onto several meaningful dissociations of model parameters.


Cortical and spinal excitabilities are differently balanced in power athletes.
It is recognised that power-sport practices have a particular effect on lower-limb neuromuscular parameters. Less is known about corticospinal network adaptation, however, or whether these adaptations are specific to the lower limb. In the present study, the corticospinal and spinal excitabilities of upper and lower limbs have been examined in a group of untrained participants (UT, n = 10) and compared to those of a group of well-trained athletes practicing parkour (PK, n = 10). This activity, consisting of overcoming obstacles offered by the urban environment, was chosen as a model of power activity. The motor evoked potentials (MEPs) induced by transcranial magnetic stimulations and H-reflexes and maximal M-waves evoked by peripheral nerve stimulations were elicited in both upper- (flexor carpi radialis [FCR]) and lower-limb muscles (soleus [SOL] and gastrocnemius medialis [GM]). The results tended toward an overall greater corticospinal excitability in PK than in UT (as evidenced by greater MEP/Mmax ratio) and lower spinal excitability (lower Hmax/Mmax). H/MMAX ratio was lower for PK (0.32) than for UT (0.41) in SOL (p = 0.02), while MEP/MMAX was greater for PK than for UT in FCR (PK: 0.12; UT: 0.06; P = 0.04) and in GM (PK: 0.05, UT: 0.03, P = 0.02). In both limbs, the decrease of spinal excitability induced by parkour practice was counterbalanced by an increase in cortical excitability. Finally, the present study indicates that such long-term power practice leads to similar corticospinal plasticity in upper and lower limbs, explained by the similar solicitation of those muscles.


Antibiotic effectiveness: balancing conservation against innovation.
Antibiotic effectiveness is a natural societal resource that is diminished by antibiotic use. As with other such assets, keeping it available requires both conservation and innovation. Conservation encompasses making the best use of current antibiotic effectiveness by reducing demand through vaccination, infection control, diagnostics, public education, incentives for clinicians to prescribe fewer antibiotics, and restrictions on access to newer, last-resort antibiotics. Innovation includes improving the efficacy of current drugs and replenishing effectiveness by developing new drugs. In this paper, I assess the relative benefits and costs of these two approaches to maintaining our ability to treat infections.


Severe hypoglycaemia associated with ingesting counterfeit medication.
Cross-border importation of traditional and prescription medications is common, and many of these drugs are not approved by the Australian Therapeutic Goods Administration. Furthermore, counterfeit versions of prescription medications are also available (eg, weight-loss medications, anabolic steroids, and medications to enhance sexual performance). We describe a 54-year-old man with the first Australian case of severe hypoglycaemia induced by imported, laboratory-confirmed counterfeit Cialis. This serves to remind medical practitioners that counterfeit medication may be the cause of severe hypoglycaemia (or other unexplained illness).


Group-specific 16S rRNA targeted probes for the detection of type I and type II methanotrophs by fluorescence in situ hybridisation.
The study of methane-oxidising bacteria (methanotrophs) is of special interest, because of their role in the natural reduction of methane emissions from many different sources. Therefore new probes were developed to detect specifically either type I (Methylococcaceae) or type II methanotrophs (Methylocystaceae). The probes have shown high specificity in fluorescence in situ hybridisations (FISH), as demonstrated by parallel hybridisation of target and reference strains as well as sequence data analysis. With these probes, methanotrophs were detected in soil and root samples from rice microcosms, demonstrating their applicability even in a complex environmental matrix.


Visual attention and dietary supplementation in children with perinatal brain injury.
To investigate whether children with perinatal brain injury have impairments in specific components of visual attention, and whether early dietary supplementation can reduce any deficits. Children participating in the Dolphin neonatal trial of dietary supplementation were tested at age 6 months with the Infant Fixation Shift Attention Test, and at 4 to 5 years with four subtests of the Early Childhood Attention Battery (ECAB) assessing different components of attention (selective, sustained, and executive function), and the Fluid Crystallized Intelligence Index of the Kaufman Assessment Battery for Children, Second Edition (KABC-II). From 59 children originally assigned to trial groups, 33 were available for testing at 4 to 5 years (18 treatment group of whom seven, six, and five showed mild, moderate, or severe neonatal brain injury; 15 controls with one, seven, and seven in the neonatal brain injury categories respectively). Given the imbalance in numbers with mild brain injury, analysis of trial group differences is restricted to moderate and severe brain injury severities (n=25). Children with perinatal brain injury showed poorer attention across all components relative to age norms (mean standard scores 75-87; p<0.001 for three of the four subtests), with the greatest impairment in sustained attention. These impairments remained when compared with cognitive age assessed using the Fluid Crystallized Intelligence Index. Impairment was reduced in the treatment compared to the control group (p=0.04 for flanker test, p=0.002 for counterpointing, and p=0.027 for the overall ECAB score). Perinatal brain injury is associated with later impaired attention, beyond that predicted from any general cognitive disability. Impairment varies across attention components, being most severe for sustained attention. The effects on flanker and counterpointing suggest that dietary supplementation from 0 to 2 years of age may reduce attention problems. Measuring the different components of attention is important when considering assessment and interventions for children with perinatal brain injury.


Novel haem co-ordination variants of flavocytochrome P450BM3.
Bacillus megaterium flavocytochrome P450 BM3 is a catalytically self-sufficient fatty acid hydroxylase formed by fusion of soluble NADPH-cytochrome P450 reductase and P450 domains. Selected mutations at residue 264 in the haem (P450) domain of the enzyme lead to novel amino acid sixth (distal) co-ordination ligands to the haem iron. The catalytic, spectroscopic and thermodynamic properties of the A264M, A264Q and A264C variants were determined in both the intact flavocytochromes and haem domains of P450 BM3. Crystal structures of the mutant haem domains demonstrate axial ligation of P450 haem iron by methionine and glutamine ligands trans to the cysteine thiolate, creating novel haem iron ligand sets in the A264M/Q variants. In contrast, the crystal structure of the A264C variant reveals no direct interaction between the introduced cysteine side chain and the haem, although EPR data indicate Cys(264) interactions with haem iron in solution. The A264M haem potential is elevated by comparison with wild-type haem domain, and substrate binding to the A264Q haem domain results in a approximately 360 mV increase in potential. All mutant haem domains occupy the conformation adopted by the substrate-bound form of wild-type BM3, despite the absence of added substrate. The A264M mutant (which has higher dodecanoate affinity than wild-type BM3) co-purifies with a structurally resolved lipid. These data demonstrate that a single mutation at Ala(264) is enough to perturb the conformational equilibrium between substrate-free and substrate-bound P450 BM3, and provide firm structural and spectroscopic data for novel haem iron ligand sets unprecedented in nature.


Cerebrospinal meningitis with the presence of Acinetobacter spp.
The study was conducted on 16 strains of Acinetobacter sp. which were isolated from cerebrospinal fluid. The diagnostic material was analysed with the use of automatic BacT/Alert system (Organon Teknika). The analysis was performed in the Department of Microbiology, Medical University in Bydgoszcz. API 20NE system (bioMérieux) enabled the identification of 14 strains (87.5%) as A. baumannii, 1 strain as A. haemolyticus and 1 strain as A. lwoffii. The micro-organisms were isolated from patients whose age ranged between 4 and 66 years. These patients were treated in the departments of Neurosurgery (75.0%), Neurology (18.8%) and Intensive Therapy (6.2%). The infection of cerebrospinal fluid was caused by injury and subsequent exposure to the bacteria present in external environment. Antibiotic-sensitivity of these micro-organisms was evaluated with the help of disc-diffusion method, observing standardisation conditions outlined by NCCLS. All the strains proved sensitive to carbapenems, 15 strains were sensitive to netilmicin, 7 strains--to tobramycin and 7 strains--to amikacin. All the strains displayed multiple resistance. The only exception was A. haemolyticus. The use of two-discs allowed for the detection of ESBLs in 7 A. baumannii strains. Positive results were most frequently obtained after the combination of sublactam and aztreonam. Due to microscopic resemblance between Acinetobacter spp., and bacteria of Neisseria, Moraxella and Haemophilus genus, microbiological diagnostics should not be restricted to microscopic assessment of cerebrospinal fluid and quick serological tests evaluating the antigens of the most frequent aetiological factors. Considering multiple resistance of Acinetobacter spp. to antibiotics, the treatment should be based on sensitivity tests and the ability of a given antibiotic to penetrate into cerebrospinal fluid. In our opinion, both reasonable antibiotic policy as well as observing the principles of hygiene and monitoring infections play equally important roles in the prevention of infections with Acinetobacter spp. Such combined measures may help to prevent the spreading of multiple resistant strains in hospital environment.


Identification of a Novel O-Conotoxin Reveals an Unusual and Potent Inhibitor of the Human α9α10 Nicotinic Acetylcholine Receptor.
Conotoxins are a pool of disulfide-rich peptide neurotoxins produced by cone snails for predation and defense. They are a rich reservoir of novel ligands for ion channels, neurotransmitter receptors and transporters in the nervous system. In this study, we identified a novel conotoxin component, O-conotoxin GeXXVIIA, from the venom of Conus generalis. The native form of this component is a disulfide-linked homodimer of a 5-Cys-containing peptide. Surprisingly, our electrophysiological studies showed that, in comparison to the folded monomers, the linear peptide of this toxin had the highest inhibitory activity at the human α9α10 nicotinic acetylcholine receptor (nAChR), with an IC50 of 16.2 ± 1.4 nM. The activities of the N-terminal and C-terminal halves of the linear toxin are markedly reduced compared with the full-length toxin, suggesting that the intact sequence is required to potently inhibit the hα9α10 nAChR. α9α10 nAChRs are expressed not only in the nervous system, but also in a variety of non-neuronal cells, such as cochlear hair cells, keratinocytes, epithelial and immune cells. A potent inhibitor of human α9α10 nAChRs, such as GeXXVIIA, would facilitate unraveling the functions of this nAChR subtype. Furthermore, this unusual nAChR inhibitor may lead to the development of novel α9α10 nAChR-targeting drugs.


Nebulised fentanyl for post-operative pain relief, a prospective double-blind controlled randomised clinical trial.
Intravenous (IV) route for fentanyl administration is the gold standard for post-operative pain relief, but complications such as respiratory depression, bradycardia and hypotension have limited this route. The aim of this randomised controlled trial was to compare the efficacy of nebulised fentanyl with IV fentanyl for post-operative pain relief after lower abdominal surgery. In the post-operative care unit, at the time of first onset of pain (visual analogue scale- VAS score > 4) patients were randomised into three groups and fentanyl was administered either IV 2 μg/kg or by nebulisation of solution containing 3 or 4 μg/kg fentanyl over 8 min in 90 patients divided into three groups of 30 each. Observation were made for pain relief by visual analogue scale score 0-10. Adverse effects such as respiratory depression, bradycardia and hypotension were also recoded. Statistical analysis was performed using Medcalc software version 12, 2012. (MedCalc Software, Ostend, Belgium). In the nebulisation group, it was observed that the analgesic efficacy of fentanyl was dose dependent with a delayed onset of analgesia (10 min vs. 5 min). Nebulisation with 4 μg/kg fentanyl produced analgesia at par to 2 μg/kg IV fentanyl with prolonged duration (90 min vs. 30 min) and with significantly less adverse effects. This study shows that nebulisation with 4 μg/kg fentanyl may be used as an alternative to IV 2 μg/kg fentanyl for adequate post-operative pain relief.


A dual role for homothorax in inhibiting wing blade development and specifying proximal wing identities in Drosophila.
The Drosophila wing imaginal disc gives rise to three body parts along the proximo-distal (P-D) axis: the wing blade, the wing hinge and the mesonotum. Development of the wing blade initiates along part of the dorsal/ventral (D/V) compartment boundary and requires input from both the Notch and wingless (wg) signal transduction pathways. In the wing blade, wg activates the gene vestigial (vg), which is required for the wing blade to grow. wg is also required for hinge development, but wg does not activate vg in the hinge, raising the question of what target genes are activated by wg to generate hinge structures. Here we show that wg activates the gene homothorax (hth) in the hinge and that hth is necessary for hinge development. Further, we demonstrate that hth also limits where along the D/V compartment boundary wing blade development can initiate, thus helping to define the size and position of the wing blade within the disc epithelium. We also show that the gene teashirt (tsh), which is coexpressed with hth throughout most of wing disc development, collaborates with hth to repress vg and block wing blade development. Our results suggest that tsh and hth block wing blade development by repressing some of the activities of the Notch pathway at the D/V compartment boundary.


Soccer vs. running training effects in young adult men: which programme is more effective in improvement of body composition? Randomized controlled trial.
The aims of this study were: 1) To determine the effects of a 12-week recreational soccer training programme and continuous endurance running on body composition of young adult men and 2) to determine which of these two programmes was more effective concerning body composition. Sixty-four participants completed the randomized controlled trial and were randomly assigned to one of three groups: a soccer training group (SOC; n=20), a running group (RUN; n=21) or a control group performing no physical training (CON; n=23). Training programmes for SOC and RUN lasted 12-week with 3 training sessions per week. Soccer sessions consisted of 60 min ordinary five-a-side, six-a-side or seven-a-side matches on a 30-45 m wide and 45-60 m long plastic grass pitch. Running sessions consisted of 60 min of continuous moderate intensity running at the same average heart rate as in SOC (~80% HRmax). All participants, regardless of group assignment, were tested for each of the following dependent variables: body weight, body height, body mass index, percent body fat, body fat mass, fat-free mass and total body water. In the SOC and RUN groups there was a significant decrease (p < 0.05) in body composition parameters from pre- to post-training values for all measures with the exception of fat-free mass and total body water. Body mass index, percent body fat and body fat mass did not differ between groups at baseline, but by week 12 were significantly lower (p < 0.05) in the SOC and RUN groups compared to CON. To conclude, recreational soccer training provides at least the same changes in body composition parameters as continuous running in young adult men when the training intensity is well matched.


Methods for the Study of Verticillium chlamydosporium in the Rhizosphere.
Methods for screening isolates of the nematophagous fungus, V. chlamydosporium, for their ability to colonize the surface of plant roots are described. Significant differences in the extent of colonization were observed in sterile conditions and in soil; plant species and cultivars also differed in their ability to support a selected isolate of the fungus. Although fungal density could be estimated using a semi-selective medium, it was not possible to separate differences in vegetative growth from differences in sporulation. There was a weak positive correlation between estimates of fungal density on the roots of plants grown in sterile conditions and the extent of hyphal growth measured by direct observation.


iSEE: Interactive SummarizedExperiment Explorer.
Data exploration is critical to the comprehension of large biological data sets generated by high-throughput assays such as sequencing. However, most existing tools for interactive visualisation are limited to specific assays or analyses. Here, we present the iSEE (Interactive SummarizedExperiment Explorer) software package, which provides a general visual interface for exploring data in a SummarizedExperiment object. iSEE is directly compatible with many existing R/Bioconductor packages for analysing high-throughput biological data, and provides useful features such as simultaneous examination of (meta)data and analysis results, dynamic linking between plots and code tracking for reproducibility. We demonstrate the utility and flexibility of iSEE by applying it to explore a range of real transcriptomics and proteomics data sets.


Evaluation of oxidant stress and antioxidant defense in discrete brain regions of rats exposed to 900 MHz radiation.
In the current study, the effects of 900 MHz radio-frequency electromagnetic radiation (RF-EMR) on levels of thiobarbituric acid-reactive substances (TBARS), total antioxidants (TA), and glutathione S-transferase (GST) activity in discrete brain regions were studied in adolescent rats. Thirty-six male Wistar rats (6-8 weeks old) were allotted into three groups (n = 12 in each group). Control group (1) remained undisturbed in their home cage; sham group (2) was exposed to mobile phone in switch off mode for four weeks; RF-EMR-exposed group (3) was exposed to 900 MHz of RF-EMR (1 hr/day with peak power density of 146.60 µW/cm2) from an activated Global System for Mobile communication (GSM) mobile phone (kept in silent mode; no ring tone and no vibration) for four weeks. On 29th day, behavioral analysis was done. Followed by this, six animals from each group were sacrificed and biochemical parameters were studied in amygdala, hippocampus, frontal cortex, and cerebellum. Altered behavioral performances were found in RF-EMR-exposed rats. Additionally, elevated TBARS level was found with all brain regions studied. RF-EMR exposure significantly decreased TA in the amygdala and cerebellum but its level was not significantly changed in other brain regions. GST activity was significantly decreased in the hippocampus but, its activity was unaltered in other brain regions studied. RF-EMR exposure for a month induced oxidative stress in rat brain, but its magnitude was different in different regions studied. RF-EMR-induced oxidative stress could be one of the underlying causes for the behavioral deficits seen in rats after RF-EMR exposure (Fig. 5, Ref. 37).


NMN transport by snake renal tubules: choline effects, countertransport, H+-NMN exchange.
N1-methylnicotinamide (NMN) transport by isolated perfused snake (Thamnophis spp.) proximal renal tubules was examined for effects of choline and evidence of countertransport of organic cations and H+-NMN exchange. Choline had no effect on NMN transport, supporting the concept of multiple organic cation transport processes. A transconcentration of unlabeled NMN had no effect on unidirectional transepithelial fluxes of labeled NMN. Low pH in the bath enhanced the lumen-to-bath flux of labeled NMN and, at the same time, reduced the intracellular concentration of labeled NMN, but low pH in the perfusate had no effect on the bath-to-lumen flux of labeled NMN. To examine transport at single membranes more directly in intact tubules, we examined efflux of labeled NMN across the luminal membrane of tubules covered with oil and across the peritubular membrane of tubules with oil-filled lumens. Neither unlabeled NMN nor low pH in the perfusate had any effect on efflux across the luminal membrane, providing no evidence of countertransport or H+-NMN exchange across this membrane. However, unlabeled NMN and low pH in the bath both significantly stimulated efflux across the peritubular membrane, providing evidence that both countertransport of organic cations and H+-NMN exchange may play a role in transport of NMN across this membrane.


The effects of QEEG-informed neurofeedback in ADHD: an open-label pilot study.
In ADHD several EEG biomarkers have been described before, with relevance to treatment outcome to stimulant medication. This pilot-study aimed at personalizing neurofeedback treatment to these specific sub-groups to investigate if such an approach leads to improved clinical outcomes. Furthermore, pre- and post-treatment EEG and ERP changes were investigated in a sub-group to study the neurophysiological effects of neurofeedback. Twenty-one patients with ADHD were treated with QEEG-informed neurofeedback and post-treatment effects on inattention (ATT), hyperactivity/impulsivity (HI) and comorbid depressive symptoms were investigated. There was a significant improvement for both ATT, HI and comorbid depressive complaints after QEEG-informed neurofeedback. The effect size for ATT was 1.78 and for HI was 1.22. Furthermore, anterior individual alpha peak frequency (iAPF) demonstrated a strong relation to improvement on comorbid depressive complaints. Pre- and post-treatment effects for the SMR neurofeedback sub-group exhibited increased N200 and P300 amplitudes and decreased SMR EEG power post-treatment. This pilot study is the first study demonstrating that it is possible to select neurofeedback protocols based on individual EEG biomarkers and suggests this results in improved treatment outcome specifically for ATT, however these results should be replicated in further controlled studies. A slow anterior iAPF at baseline predicts poor treatment response on comorbid depressive complaints in line with studies in depression. The effects of SMR neurofeedback resulted in specific ERP and EEG changes.


Endothelin: an important factor in acute renal failure?
Very low concentrations of the vasoconstrictor peptide endothelin cause intense long-lasting renal vasoconstriction. In the isolated perfused rat kidney, the concentration of endothelin required to reduce blood-flow by 50% is 200 pmol/l, compared with 1000 pmol/l angiotensin II (previously the most potent known vasoconstrictor). Whereas angiotensin II has little effect on the glomerular filtration rate (GFR), a rise in endothelin from 100 to 800 pmol/l reduces GFR by 90%. Endothelin is probably present in the circulation at low concentrations in vivo; events associated clinically with acute renal failure would tend to increase this concentration. Endothelin may be a mediator in the pathogenesis of acute renal failure.


Pairwise likelihood estimation for confirmatory factor analysis models with categorical variables and data that are missing at random.
Methods for the treatment of item non-response in attitudinal scales and in large-scale assessments under the pairwise likelihood (PL) estimation framework and under a missing at random (MAR) mechanism are proposed. Under a full information likelihood estimation framework and MAR, ignorability of the missing data mechanism does not lead to biased estimates. However, this is not the case for pseudo-likelihood approaches such as the PL. We develop and study the performance of three strategies for incorporating missing values into confirmatory factor analysis under the PL framework, the complete-pairs (CP), the available-cases (AC) and the doubly robust (DR) approaches. The CP and AC require only a model for the observed data and standard errors are easy to compute. Doubly-robust versions of the PL estimation require a predictive model for the missing responses given the observed ones and are computationally more demanding than the AC and CP. A simulation study is used to compare the proposed methods. The proposed methods are employed to analyze the UK data on numeracy and literacy collected as part of the OECD Survey of Adult Skills.


Determinants of differential HIV incidence among women in three southern African locations.
We explored factors associated with differential HIV incidence among women participating in a HIV prevention trial in Harare, Durban, and Johannesburg. The trial had shown no effect of the intervention (diaphragm and lubricant gel) on HIV incidence. A prospective cohort analysis was conducted of trial participants followed for 12-24 months. Sociodemographic, biological, and behavioral data were collected at baseline and at quarterly visits. Factors associated with HIV incidence were estimated using multivariable Cox regression models, stratified by study location. Attributable risk was calculated from the adjusted hazard ratios (AHR). There were 309 incident HIV infections among the 4948 women in the analysis. HIV incidence was highest in Durban [6.75/100 person-years; 95% confidence interval (CI): 5.74 to 7.93], lower in Johannesburg (3.33/100 person-years; 95% CI: 2.51 to 4.44), and lowest in Harare (2.72/100 person-years; 95% CI: 2.26 to 3.26). Sexually transmitted infections were important risk factors in Harare [prevalent herpes simplex virus type 2 (HSV2) AHR = 2.56, 95% CI: 1.61 to 4.06; incident HSV2 AHR = 12.60, 95% CI: 2.13 to 21.87; Neisseria gonorrhoeae AHR = 6.82, 95% CI: 2.13 to 21.87] and in Durban (prevalent HSV2 AHR = 1.64, 95% CI: 1.07 to 2.51; N. gonorrhoeae AHR = 4.40, 95% CI: 2.07 to 9.39). In Durban, having multiple partners (adjusted odds ratio (AOR) = 1.78 95% CI: 1.11 to 2.85) and sex although a partner was under the influence of alcohol/drugs (AOR = 1.51 95% CI: 1.05 to 2.16) significantly increased risk, whereas in Johannesburg, sexual debut <16 years (AOR = 2.60 95% CI: 1.30 to 5.17) was a strong predictor of HIV acquisition. Important differences were seen in drivers of HIV incidence at the 3 study locations. Results from this analysis imply that targeted HIV programing could have a large impact on incident HIV infection in women, and that the most effective approach will likely vary based on knowledge of the local situation/epidemiology.


An Unsettled Debate About the Potential Role of Infection in Pathogenesis of Atherosclerosis.
Association of infection with atherosclerosis is by no means new. Several sero-epidemiological and pathologic studies as well as animal models have shown a link between infection and atherosclerosis. Exciting discoveries in recent times related to role of inter-individual genetic variation in modulating inflammatory response to infection have reignited the enthusiasm in proving a causal link between infection and atherosclerosis. The purpose of this article was to review and analyze the available evidence linking infection with atherosclerosis.


Association mapping and genetic dissection of drought-induced canopy temperature differences in rice.
Drought-stressed plants display reduced stomatal conductance, which results in increased leaf temperature by limiting transpiration. In this study, thermal imaging was used to quantify the differences in canopy temperature under drought in a rice diversity panel consisting of 293 indica accessions. The population was grown under paddy field conditions and drought stress was imposed for 2 weeks at flowering. The canopy temperature of the accessions during stress negatively correlated with grain yield (r= -0.48) and positively with plant height (r=0.56). Temperature values were used to perform a genome-wide association (GWA) analysis using a 45K single nucleotide polynmorphism (SNP) map. A quantitative trait locus (QTL) for canopy temperature under drought was detected on chromosome 3 and fine-mapped using a high-density imputed SNP map. The candidate genes underlying the QTL point towards differences in the regulation of guard cell solute intake for stomatal opening as the possible source of temperature variation. Genetic variation for the significant markers of the QTL was present only within the tall, low-yielding landraces adapted to drought-prone environments. The absence of variation in the shorter genotypes, which showed lower leaf temperature and higher grain yield, suggests that breeding for high grain yield in rice under paddy conditions has reduced genetic variation for stomatal response under drought.


Brain ageing in schizophrenia: evidence from 26 international cohorts via the ENIGMA Schizophrenia consortium.
Schizophrenia (SZ) is associated with an increased risk of life-long cognitive impairments, age-related chronic disease, and premature mortality. We investigated evidence for advanced brain ageing in adult SZ patients, and whether this was associated with clinical characteristics in a prospective meta-analytic study conducted by the ENIGMA Schizophrenia Working Group. The study included data from 26 cohorts worldwide, with a total of 2803 SZ patients (mean age 34.2 years; range 18-72 years; 67% male) and 2598 healthy controls (mean age 33.8 years, range 18-73 years, 55% male). Brain-predicted age was individually estimated using a model trained on independent data based on 68 measures of cortical thickness and surface area, 7 subcortical volumes, lateral ventricular volumes and total intracranial volume, all derived from T1-weighted brain magnetic resonance imaging (MRI) scans. Deviations from a healthy brain ageing trajectory were assessed by the difference between brain-predicted age and chronological age (brain-predicted age difference [brain-PAD]). On average, SZ patients showed a higher brain-PAD of +3.55 years (95% CI: 2.91, 4.19; I2 = 57.53%) compared to controls, after adjusting for age, sex and site (Cohen's d = 0.48). Among SZ patients, brain-PAD was not associated with specific clinical characteristics (age of onset, duration of illness, symptom severity, or antipsychotic use and dose). This large-scale collaborative study suggests advanced structural brain ageing in SZ. Longitudinal studies of SZ and a range of mental and somatic health outcomes will help to further evaluate the clinical implications of increased brain-PAD and its ability to be influenced by interventions.


A hybrid micro-macroevolutionary approach to gene tree reconstruction.
Gene family evolution is determined by microevolutionary processes (e.g., point mutations) and macroevolutionary processes (e.g., gene duplication and loss), yet macroevolutionary considerations are rarely incorporated into gene phylogeny reconstruction methods. We present a dynamic program to find the most parsimonious gene family tree with respect to a macroevolutionary optimization criterion, the weighted sum of the number of gene duplications and losses. The existence of a polynomial delay algorithm for duplication/loss phylogeny reconstruction stands in contrast to most formulations of phylogeny reconstruction, which are NP-complete. We next extend this result to obtain a two-phase method for gene tree reconstruction that takes both micro- and macroevolution into account. In the first phase, a gene tree is constructed from sequence data, using any of the previously known algorithms for gene phylogeny construction. In the second phase, the tree is refined by rearranging regions of the tree that do not have strong support in the sequence data to minimize the duplication/lost cost. Components of the tree with strong support are left intact. This hybrid approach incorporates both micro- and macroevolutionary considerations, yet its computational requirements are modest in practice because the two-phase approach constrains the search space. Our hybrid algorithm can also be used to resolve nonbinary nodes in a multifurcating gene tree. We have implemented these algorithms in a software tool, NOTUNG 2.0, that can be used as a unified framework for gene tree reconstruction or as an exploratory analysis tool that can be applied post hoc to any rooted tree with bootstrap values. The NOTUNG 2.0 graphical user interface can be used to visualize alternate duplication/loss histories, root trees according to duplication and loss parsimony, manipulate and annotate gene trees, and estimate gene duplication times. It also offers a command line option that enables high-throughput analysis of a large number of trees.


Detection of human retrovirus 5 in patients with arthritis and systemic lupus erythematosus.
To examine whether human retrovirus 5 (HRV-5) infection is associated with autoimmune rheumatic disease. DNA from patients with various disorders including inflammatory diseases and from normal subjects was tested by nested polymerase chain reaction (PCR) for HRV-5 proviral DNA. Positive results were confirmed by DNA sequencing. HRV-5 proviral DNA was detected in 53% of synovial samples from arthritic joints, in 12% of blood samples from patients with rheumatoid arthritis (RA), and in 16% of blood samples from patients with systemic lupus erythematosus. In contrast, it was not detectable by PCR of affected tissues from patients with several other autoimmune diseases and was found in only 1 of >200 tissue specimens obtained at autopsy from non-RA patients. Sequence analysis of the amplified viral segment showed genetic variation between samples with maintenance of the open reading frame, typical of a replicating infectious retrovirus. This is the first report of the frequent detection of HRV-5 in any disease. We propose that the possible involvement of HRV-5 in autoimmune and rheumatic disease should be investigated further.


Simian virus 40 large-T-antigen-specific rejection of mKSA tumor cells in BALB/c mice is critically dependent on both strictly tumor-associated, tumor-specific CD8(+) cytotoxic T lymphocytes and CD4(+) T helper cells.
Protective immunity of BALB/c mice immunized with simian virus 40 (SV40) large T antigen (TAg) against SV40-transformed, TAg-expressing mKSA tumor cells is critically dependent on both CD8(+) and CD4(+) T lymphocytes. By depleting mice of T-cell subsets at different times before and after tumor challenge, we found that at all times, CD4(+) and CD8(+) cells both were equally important in establishing and maintaining a protective immune response. CD4(+) cells do not contribute to tumor eradication by directly lysing mKSA cells. However, CD4(+) lymphocytes provide help to CD8(+) cells to proliferate and to mature into fully active cytotoxic T lymphocytes (CTL). Depletion of CD4(+) cells by a single injection of CD4-specific monoclonal antibody at any time from directly before injection of the vaccinating antigen to up to 7 days after tumor challenge inhibited the generation of cytolytic CD8(+) lymphocytes. T helper cells in this system secrete the typical Th-1 cytokines interleukin 2 (IL-2) and gamma interferon. Because in this system TAg-specific CD8(+) cells secrete only minute amounts of IL-2, it appears that T helper cells provide these cytokines for CD8(+) T cells. Moreover, this helper effect of CD4(+) T cells in mKSA tumor rejection in BALB/c mice does not simply improve the activity of TAg-specific CD8(+) CTL but actually enables them to mature into cytolytic effector cells. Beyond this activity, the presence of T helper cells is necessary even in the late phase of tumor cell rejection in order to maintain protective immunity. However, despite the support of CD4(+) T helper cells, the tumor-specific CTL response is so weak that only at the site of tumor cell inoculation and not in the spleen or in the regional lymph nodes can TAg-specific CTL be detected.


WE-D-218-01: Ultrasound Scanner Innovations and Clinical Practice.
Of all the imaging modalities, ultrasound scanners have gone through the most profound changes over the last several decades in terms of their size, capability, and cost. Much of this is due to the small data acquisition devices (ultrasound transducers) and Moore's Law dependent signal/image processors that comprise and ultrasound scanner. These are in direct contrast with the front ends of MRI or CT scanners with their sizeable power hungry gantries. Thus ultrasound has been a direct beneficiary of the miniaturization associated with the semiconductor industry; this has enabled the migration of much hardware functionality to software and development of much smaller devices even including handheld scanners. Such changes are having a significant impact on clinical utilization of ultrasound. This talk will review some of these including the recent introduction of complete software backends, i.e. ultrasound scanners composed of analog front ends which are connected to processors with minimal dedicated digital hardware. 1. Understand the architecture of an ultrasound scanner and how it has changed with evolving technology. 2. Understand the implications to clinical practice from these changes. 3. Understand the possibilities for the future of ultrasound scanners both from the view of new technical capabilities and how these might impact the clinic.


The Nature, Extent, and Consequences of Genetic Variation in the opa Repeats of Notch in Drosophila.
Polyglutamine (pQ) tracts are abundant in proteins co-interacting on DNA. The lengths of these pQ tracts can modulate their interaction strengths. However, pQ tracts >40 residues are pathologically prone to amyloidogenic self-assembly. Here, we assess the extent and consequences of variation in the pQ-encoding opa repeats of Notch in Drosophila melanogaster. We use Sanger sequencing to genotype opa sequences ([Formula: see text]-CAX repeats), which have resisted assembly using short sequence reads. While most sampled lines carry the major allele opa31 encoding Q13HQ17 or the opa32 allele encoding Q13HQ18, many lines carry rare alleles encoding pQ tracts >32 residues: opa33a (Q14HQ18), opa33b (Q15HQ17), opa34 (Q16HQ17), opa35a1/opa35a2 (Q13HQ21), opa36 (Q13HQ22), and opa37 (Q13HQ23). Only one rare allele encodes a tract <31 residues: opa23 (Q13-Q10). This opa23 allele shortens the pQ tract while simultaneously eliminating the interrupting histidine. We introgressed these opa variant alleles into common backgrounds and measured the frequency of Notch-type phenotypes. Homozygotes for the short and long opa alleles have defects in embryonic survival and sensory bristle organ patterning, and sometimes show wing notching. Consistent with functional differences between Notch opa variants, we find that a scute inversion carrying the rare opa33b allele suppresses the bristle patterning defect caused by achaete/scute insufficiency, while an equivalent scute inversion carrying opa31 manifests the patterning defect. Our results demonstrate the existence of potent pQ variants of Notch and the need for long read genotyping of key repeat variables underlying gene regulatory networks.


Favorable treatment outcome in non-Hodgkin's lymphoma patients with "poor" mobilization of peripheral blood progenitor cells.
Our purpose was to evaluate the outcome and costs of high-dose chemotherapy and autologous peripheral blood progenitor cell (PBPC) transplantation in patients with the inability to mobilize sufficient numbers of PBPCs to allow rapid engraftment after PBPC transplantation. We treated 172 consecutive non-Hodgkin's lymphoma (NHL) patients with cyclophosphamide and granulocyte colony-stimulating factor followed by apheresis to collect PBPCs. The cells were separated on a Percoll gradient and purged with monoclonal antibodies and complement. The patients were categorized as "good" mobilizers if a collection of > or =2 x 10(6) CD34+ cells/kg was obtained (n = 138, 80%) or "poor" mobilizers if <2 x 10(6) CD34+ cells/kg were obtained (n = 34, 20%). With a median follow-up of 3.5 years, there is no statistically significant difference in actuarial event-free survival, overall survival, or relapse for good mobilizers compared with poor mobilizers. However, there was a trend toward increasing nonrelapse, transplantation-related mortality of 11.8% for poor mobilizers versus 3.6% for good mobilizers (P = .08) and early death from all causes including relapse within 120 days (poor 20.6% versus good 8.7%, P = .06). The total cost for bone marrow transplantation-related care was significantly higher, at $140,264 for poor mobilizers versus $80,833 for good mobilizers (P = .0001). The population of patients with NHL who mobilize PBPCs poorly into the circulation have a higher cost for posttransplant support. However, there is no significant difference in relapse, event-free survival, or overall survival for such patients compared with those who mobilize PBPCs easily.


Alectinib Resistance in ALK-Rearranged Lung Cancer by Dual Salvage Signaling in a Clinically Paired Resistance Model.
The mechanisms responsible for the development of resistance to alectinib, a second-generation anaplastic lymphoma kinase (ALK) inhibitor, are still unclear, and few cell lines are currently available for investigating ALK-rearranged lung cancer. To identify the mechanisms underlying acquired resistance to alectinib, two patient-derived cell lines were established from an alectinib-naïve ALK-rearranged lung cancer and then after development of alectinib resistance. The properties acquired during treatments were detected by comparisons of the two cell lines, and then functional analyses were performed. Coactivation of c-Src and MET was identified after the development of alectinib resistance. Combinatorial therapy against Src and MET significantly restored alectinib sensitivity in vitro (17.2-fold). Increased apoptosis, reduction of tumor volume, and inhibition of MAPK and PI3K/AKT signaling molecules for proliferation and survival were observed when the three kinases (Src, MET, and ALK) were inhibited. A patient-derived xenograft from the alectinib-resistant cells indicated that combination therapy with a saracatinib and crizotinib significantly decreased tumor size in vivo. To confirm the generality, a conventional alectinib-resistant cell line model (H2228-AR1S) was established from NCI-H2228 cells (EML4-ALK variant 3a/b). In H2228-AR1S, combination inhibition of Src and MET also restored alectinib sensitivity. These data reveal that dual salvage signaling from MET and Src is a potential therapeutic target in alectinib-resistant patients. IMPLICATIONS: This study demonstrates the feasibility to elucidate personalized drug-resistance mechanisms from individual patient samples.


Evaluation of emotion-based messages designed to motivate Hispanic and Asian parents of early adolescents to engage in calcium-rich food and beverage parenting practices.
Setting healthful beverage expectations, making calcium-rich foods and beverages (CRF/B) available, and role modeling are parenting practices promoting calcium intake among early adolescents. This study aimed to evaluate emotion-based messages designed to motivate parents of early adolescents to perform these practices. Emotion-based messages were developed for each parenting practice and tested in 35 parents from 5 states. Findings were used to modify messages and develop a survey administered via Amazon MechanicalTurk to a convenience sample of Asian (n = 166) and Hispanic (n = 184) parents of children 10-13 years. Main outcome measures were message comprehension, motivation, relevance, acceptability, and novelty. Engagement in the parenting practices was also assessed. Message comprehension was acceptable for the majority of parents. Most also agreed that messages were motivational (setting healthful beverage expectations (69.0%), making CRF/B available (67.4%), and role modeling (80.0%)), relevant and acceptable. About 30-50% indicated they had not seen the information before. Many parents indicated they were already engaging in the practices (> 70%). No racial/ethnic differences were observed for responses to messages or engaging in parenting practices. Results indicate that emotion-based messages designed to motivate parents to engage in parenting practices that promote calcium intake among early adolescents were motivating, relevant, and acceptable.


Critical assessment of the Boltzmann approach to active systems.
Generic models of propelled particle systems posit that the emergence of polar order is driven by the competition between local alignment and noise. Although this notion has been confirmed employing the Boltzmann equation, the range of applicability of this equation remains elusive. We introduce a broad class of mesoscopic collision rules and analyze the prerequisites for the emergence of polar order in the framework of kinetic theory. Our findings suggest that a Boltzmann approach is appropriate for weakly aligning systems but is incompatible with experiments on cluster forming systems.


African swine fever: Update on Eastern, Central and Southern Africa.
Control of African swine fever (ASF) in countries in Eastern, Central and Southern Africa (ECSA) is particularly complex owing to the presence of all three known epidemiological cycles of maintenance of the virus, namely an ancient sylvatic cycle involving the natural hosts and vectors of the disease as well as domestic cycles with and without involvement of natural vectors. While the situation is well documented in some of the countries, for others very little information is available. In spite of the unfavourable ASF situation, the pig population in the sub-region has grown exponentially in recent decades and is likely to continue to grow in response to rapid urban growth resulting in increasing demand for animal protein by populations that are no longer engaged in livestock production. Better management of ASF will be essential to permit the pig sector to reach its full potential as a supplier of high quality protein and a source of income to improve livelihoods and create wealth. No vaccine is currently available and it is likely that, in the near future, the sub-region will continue to rely on the implementation of preventive measures, based on the epidemiology of the disease, to avoid both the devastating losses that outbreaks can cause and the risk the sub-region poses to other parts of Africa and the world. The current situation in the ECSA sub-region is reviewed and gaps in knowledge are identified in order to support ongoing strategy development for managing ASF in endemic areas.


Interactions between two QTLs for time to anthesis on spike development and fertility in wheat.
Earliness per se (Eps) genes are reported to be important in fine-tuning flowering time in wheat independently of photoperiod (Ppd) and vernalisation (Vrn). Unlike Ppd and Vrn genes, Eps have relatively small effects and their physiological effect along with chromosomal position are not well defined. We evaluated eight lines derived from crossing two vernalisation insensitive lines, Paragon and Baj (late and early flowering respectively), to study the detailed effects of two newly identified QTLs, Eps-7D and Eps-2B and their interactions under field conditions. The effect of both QTLs was minor and was affected by the allelic status of the other. While the magnitude of effect of these QTLs on anthesis was similar, they are associated with very different profiles of pre-anthesis development which also depends on their interaction. Eps-7D affected both duration before and after terminal spikelet while not affecting final leaf number (FLN) so Eps-7D-early had a faster rate of leaf appearance. Eps-2B acted more specifically in the early reproductive phase and slightly altered FLN without affecting the leaf appearance rate. Both QTLs affected the spike fertility by altering the rate of floret development and mortality. The effect of Eps-2B was very small but consistent in that -late allele tended to produce more fertile florets.


Forskolin has a bimodal cAMP-independent effect on superoxide anion generation in isolated osteoclasts.
The mode of action of forskolin is of clinical and scientific interest since forskolin has been shown to have potentially therapeutic bone anti-resorptive and anti-hypertensive properties. Forskolin is thought to inhibit the bone resorptive activity of osteoclasts by elevating cytosolic cAMP and to mimic as well as augment the anti-resorptive effect of calcitonin (CT). Other studies have found that forskolin has a dose-dependent dual effect in mouse calavaria, stimulating bone resorption at low doses and having an inhibitory effect at high doses. However, the acute effect of forskolin on osteoclast functional modality has never been studied. The present investigation examined the effect of low (1 mM) and high doses (10 mM) of forskolin on superoxide anion (O2-) generation in isolated bone-resorbing rat osteoclasts. Forskolin was found to have a bimodal cAMP-independent effect on O2- generation, being stimulatory at a low dose and having an inhibitory effect at a higher dose. These findings also suggest that CT-induced inhibition of O2- generation in the osteoclasts is likely to be mediated by cAMP-independent pathways, perhaps involving [Ca2+]i modulation.


Age- and stress-associated C. elegans granulins impair lysosomal function and induce a compensatory HLH-30/TFEB transcriptional response.
The progressive failure of protein homeostasis is a hallmark of aging and a common feature in neurodegenerative disease. As the enzymes executing the final stages of autophagy, lysosomal proteases are key contributors to the maintenance of protein homeostasis with age. We previously reported that expression of granulin peptides, the cleavage products of the neurodegenerative disease protein progranulin, enhance the accumulation and toxicity of TAR DNA binding protein 43 (TDP-43) in Caenorhabditis elegans (C. elegans). In this study we show that C. elegans granulins are produced in an age- and stress-dependent manner. Granulins localize to the endolysosomal compartment where they impair lysosomal protease expression and activity. Consequently, protein homeostasis is disrupted, promoting the nuclear translocation of the lysosomal transcription factor HLH-30/TFEB, and prompting cells to activate a compensatory transcriptional program. The three C. elegans granulin peptides exhibited distinct but overlapping functional effects in our assays, which may be due to amino acid composition that results in distinct electrostatic and hydrophobicity profiles. Our results support a model in which granulin production modulates a critical transition between the normal, physiological regulation of protease activity and the impairment of lysosomal function that can occur with age and disease.


Elevated Epstein-Barr virus-encoded nuclear antigen-1 immune responses predict conversion to multiple sclerosis.
The aims of the study were to determine the immune responses to candidate viral triggers of multiple sclerosis (MS) in patients with clinically isolated syndromes (CISs), and to evaluate their potential value in predicting conversion to MS. Immune responses to Epstein-Barr virus (EBV), human herpesvirus 6, cytomegalovirus (HCMV), and measles were determined in a cohort of 147 CIS patients with a mean follow-up of 7 years and compared with 50 demographically matched controls. Compared with controls, CIS patients showed increased humoral (p < 0.0001) and cellular (p = 0.007) immune responses to the EBV-encoded nuclear antigen-1 (EBNA1), but not to other EBV-derived proteins. Immunoglobulin G (IgG) responses to other virus antigens and frequencies of T cells specific for HCMV and influenza virus gene products were unchanged in CIS patients. EBNA1 was the only viral antigen with which immune responses correlated with number of T2 lesions (p = 0.006) and number of Barkhof criteria (p=0.001) at baseline, and with number of T2 lesions (p = 0.012 at both 1 and 5 years), presence of new T2 lesions (p = 0.003 and p = 0.028 at 1 and 5 years), and Expanded Disability Status Scale score (p = 0.015 and p = 0.010 at 1 and 5 years) during follow-up. In a univariate Cox regression model, increased EBNA1-specific IgG responses predicted conversion to MS based on McDonald criteria (hazard ratio [95% confidence interval], 2.2 [1.2-4.3]; p = 0.003). Our results indicate that elevated immune responses toward EBNA1 are selectively increased in CIS patients and suggest that EBNA1-specific IgG titers could be used as a prognostic marker for disease conversion and disability progression.


The Use of Immersive Visualization for the Control of Dental Anxiety During Oral Debridement.
The purpose of this study was to evaluate the effects of Immersive Visualization (IV) eyewear on anxious, adult patients during oral debridement. Thirty adult volunteers (n=23 females; n=7 males) were enrolled in the study. Participants were required to be 18 years or older, exhibit at least moderate anxiety (score 9 or higher) on the Corah's Dental Anxiety Scale-Revised (DAS-R), and be generally healthy. Individuals were excluded from participation if they presented with severe dental calculus, periodontal disease, or dental caries, were taking psychotropic drugs, had a history of convulsive disorders, vertigo, or equilibrium disorders, or required antibiotic pre-medication. Subjects received a full mouth oral prophylaxis (supra- and subgingival scaling and selective polishing) by a single experienced dental hygienist. A split mouth design was utilized whereby each subject served as their own control. Subjects were randomly divided into 2 groups: Group A used IV eyewear during the first one-half of the appointment (right side of the mouth) and Group B used IV eyewear during the second one-half of the appointment (left side of the mouth). At screening, medical and dental histories were obtained, full mouth oral examinations were performed, and DAS-R was scored to determine eligibility. At baseline, the DAS-R was re-scored to validate anxiety levels. The Calmness Scale was scored pre- and post-IV treatment on a Likert scale ranging from 1 (very calm) to 7 (less calm). At the end of the study, subjects completed a Post IV Opinion survey. Data were entered into Microsoft Excel for Mac 2011 (Microsoft Corporation Version 14.3.5) and analyzed using SAS® 9.3 statistical software. Thirty subjects with a mean age of 29.9 years completed the study. Data analysis indicated no statistically significant difference between Group A and B with regard to mean DAS-R anxiety levels at baseline (3.15 and 2.40, respectively), with a p-value of 0.07. Data showed a significant difference when comparing the calmness mean scores within Group A pre- and post-IV treatments (4.66 and 2.93, respectively), with a p-value 0.01. Within Group B the data revealed a statistically significant difference between pre- and post-IV treatments (p<0.01, 4.33 and 2.13, respectively). Both treatment groups experienced a decrease in anxiety levels from pre to post IV treatments. Moreover, combined mean calmness scores of the 30 subjects (Group A and B) expressed in mean standard deviation showed there was a decrease from 4.50±1.31 in pre-IV treatment to 2.53±1.17 in post-IV treatment. Further investigation of the data showed that there was a significant correlation between calmness and gender; females reported higher levels of anxiety than men before and after IV treatment. Results from this study support the use of IV eyewear as an effective technique to reduce anxiety in adults during oral debridement. The use of the IV eyewear was well received by all subjects. The portable, affordable and easy-to-operate IV system makes this technique an appealing approach of reducing dental anxiety.


Comparison and evaluation of Renibacterium salmoninarum quantitative PCR diagnostic assays using field samples of Chinook and coho salmon.
Renibacterium salmoninarum is a Gram-positive bacterium causing bacterial kidney disease (BKD) in susceptible salmonid fishes. Several quantitative PCR (qPCR) assays to measure R. salmoninarum infection intensity have been reported, but comparison and evaluation of these assays has been limited. Here, we compared 3 qPCR primer/probe sets for detection of R. salmoninarum in field samples of naturally exposed Chinook and coho salmon first identified as positive by nested PCR (nPCR). Additional samples from a hatchery population of Chinook salmon with BKD were included to serve as strong positive controls. The 3 qPCR assays targeted either the multiple copy major soluble antigen (msa) genes or the single copy abc gene. The msa/non-fluorescent quencher (NFQ) assay amplified R. salmoninarum DNA in 53.2% of the nPCR positive samples, whereas the abc/NFQ assay amplified 21.8% of the samples and the abc/TAMRA assay 18.2%. The enzyme-linked immunosorbent assay (ELISA) successfully quantified only 16.4% of the nPCR positive samples. Although the msa/NFQ assay amplified a greater proportion of nPCR positive samples, the abc/NFQ assay better amplified those samples with medium and high ELISA values. A comparison of the geometric mean quantity ratios highlighted limitations of the assays, and the abc/NFQ assay strongly amplified some samples that were negative in other tests, in contrast to its performance among the sample group as a whole. These data demonstrate that both the msa/NFQ and abc/NFQ qPCR assays are specific and effective at higher infection levels and outperform the ELISA. However, most pathogen studies will continue to require multiple assays to both detect and quantify R. salmoninarum infection.


Effect of targeted agents on the endocrine response of breast cancer in the neoadjuvant setting: a systematic review.
Inhibition of aberrantly activated pathways cross-talking with hormone receptor (HR) improves response to endocrine therapy in patients with HR-positive advanced breast cancer. We performed a Pubmed database systematic review to ascertain the existence of a better clinical response when combining endocrine therapy with targeted agents in the neoadjuvant setting. Preclinical studies or trials evaluating toxicity were excluded. We found nine phase II trials that fulfilled the research criteria. The endocrine agents used were third generation aromatase inhibitors (AIs), anastrozole, letrozole or exemestane. The investigated targeted agents were inhibitors of tyrosine kinase receptors such as gefitinib, imatinib or trastuzumab/lapatinib, inhibitors of mTOR, such as everolimus, inhibitors of COX-2, such as celecoxib, and inhibitors of angiogenesis, such as bevacizumab. The response rate (RR) observed combining endocrine and targeted agents ranged between 36% and 90%. Overall the studies failed to show a remarkable advantage in RR in the combination group compared to historical control subjects receiving AIs alone.


Evidence that negative elongation factor represses transcription elongation through binding to a DRB sensitivity-inducing factor/RNA polymerase II complex and RNA.
Negative elongation factor (NELF) is a human transcription factor complex that cooperates with DRB sensitivity-inducing factor (DSIF)/hSpt4-hSpt5 to repress elongation by RNA polymerase II (RNAPII). NELF activity is associated with five polypeptides, including NELF-A, a candidate gene product for Wolf-Hirschhorn syndrome, and NELF-E, a putative RNA-binding protein with arginine-aspartic acid (RD) dipeptide repeats. Here we report several important findings regarding the DSIF/NELF-dependent elongation control. First, we have established an effective method for purifying the active NELF complex using an epitope-tagging technique. Second, the five polypeptides each are important and together are sufficient for its function in vitro. Third, NELF does not bind to either DSIF or RNAPII alone but does bind to the preformed DSIF/RNAPII complex. Fourth, NELF-E has a functional RNA-binding domain, whose mutations impair transcription repression without affecting known protein-protein interactions. Taken together, we propose that NELF causes RNAPII pausing through binding to the DSIF/RNAPII complex and to nascent transcripts. These results also have implications for how DSIF and NELF are regulated in a gene-specific manner in vivo.


A simple probabilistic model of multibody interactions in proteins.
Protein structure prediction methods typically use statistical potentials, which rely on statistics derived from a database of know protein structures. In the vast majority of cases, these potentials involve pairwise distances or contacts between amino acids or atoms. Although some potentials beyond pairwise interactions have been described, the formulation of a general multibody potential is seen as intractable due to the perceived limited amount of data. In this article, we show that it is possible to formulate a probabilistic model of higher order interactions in proteins, without arbitrarily limiting the number of contacts. The success of this approach is based on replacing a naive table-based approach with a simple hierarchical model involving suitable probability distributions and conditional independence assumptions. The model captures the joint probability distribution of an amino acid and its neighbors, local structure and solvent exposure. We show that this model can be used to approximate the conditional probability distribution of an amino acid sequence given a structure using a pseudo-likelihood approach. We verify the model by decoy recognition and site-specific amino acid predictions. Our coarse-grained model is compared to state-of-art methods that use full atomic detail. This article illustrates how the use of simple probabilistic models can lead to new opportunities in the treatment of nonlocal interactions in knowledge-based protein structure prediction and design.