Highlights from the 14(th) St Gallen International Breast Cancer Conference 2015 in Vienna: Dealing with classification, prognostication, and prediction refinement to personalize the treatment of patients with early breast cancer.
The refinement of the classification, the risk of relapse and the prediction of response to multidisciplinary treatment for early breast cancer has been the major theme of the 14(th) St Gallen International Breast Cancer Consensus Conference 2015. The meeting, held in Vienna, assembled 3500-4000 participants from 134 countries worldwide. It culminated, on the final day, with the International Consensus Session, delivered by 40-50 of the world's most experienced opinion leaders in the field of breast cancer treatment. The panelist addressed the "semantic" classification of breast cancer subtypes by pathology-based biomarkers (e.g. estrogen receptor, progesterone receptor and HER2) vs genomic classifiers. They also refined the biomarker prognostication dissecting the impact of the various gene signatures and pathologic variables in predicting the outcome of patients with early breast cancer in terms of early and late relapse. Finally they addressed the challenges stemming from the intra- and inter-observer variability in the assessment of pathologic variables and the role of gene signatures for the prediction of response to specific therapeutic approach such as endocrine therapy and chemotherapy and for personalizing local treatment of patients with early breast cancer. The vast majority of the questions asked during the consensus were about controversial issues. The opinion of the panel members has been used to implement guidance for treatment choice. This is the unique feature of the St. Gallen Consensus, ensuring that the resulting recommendations will take due cognizance of the variable resource limitations in different countries. Information derived from evidence based medicine and large meta-analyses is of obvious and enormous value. The weakness of this approach is that it gives particular weight to older trials (which have accumulated more event endpoints) and is frequently unable to collect sufficient detail on the patients and tumors in the trials to allow assessment of whether the treatments which are better on average offer equal value to all currently definable patient subgroups. What St Gallen can provide is clinically useful updated breast cancer treatment consensus for the majority of patients treated outside of clinical trials (>90%) in most countries.


The fate of patients with acute myeloid leukemia not undergoing induction chemotherapy.
Advances in chemotherapy for acute myeloid leukemia (AML) have resulted in the exclusion of patients not undergoing induction chemotherapy from research studies. To examine in detail the clinical experience of such patients, we retrospectively analyzed the outcomes of consecutive patients diagnosed with AML at our hospital from 2004 to 2012. Of 158 AML patients, 43 (27 %) did not undergo induction chemotherapy. Their median survival duration was 1.5 months, with 11, six, and four patients surviving more than 3, 6, and 12 months, respectively. As expected, their survival was worse than that of those treated with intensive or less-intensive induction therapy (14, 74, and 47 % at 1 year, and 0, 40, and 10 % at 4 years, respectively). Low white blood cell count at AML diagnosis and prior history of myelodysplastic syndrome were significantly associated with longer survival. Our findings suggest that modern supportive care measures do not prolong survival for AML patients not undergoing induction chemotherapy, although certain patients show relatively long survival. These data should prove helpful in discussing treatment pathways with patients for cases in which palliative or supportive therapy alone may be a viable treatment option.


Inferring signalling networks from longitudinal data using sampling based approaches in the R-package 'ddepn'.
Network inference from high-throughput data has become an important means of current analysis of biological systems. For instance, in cancer research, the functional relationships of cancer related proteins, summarised into signalling networks are of central interest for the identification of pathways that influence tumour development. Cancer cell lines can be used as model systems to study the cellular response to drug treatments in a time-resolved way. Based on these kind of data, modelling approaches for the signalling relationships are needed, that allow to generate hypotheses on potential interference points in the networks. We present the R-package 'ddepn' that implements our recent approach on network reconstruction from longitudinal data generated after external perturbation of network components. We extend our approach by two novel methods: a Markov Chain Monte Carlo method for sampling network structures with two edge types (activation and inhibition) and an extension of a prior model that penalises deviances from a given reference network while incorporating these two types of edges. Further, as alternative prior we include a model that learns signalling networks with the scale-free property. The package 'ddepn' is freely available on R-Forge and CRAN http://ddepn.r-forge.r-project.org, http://cran.r-project.org. It allows to conveniently perform network inference from longitudinal high-throughput data using two different sampling based network structure search algorithms.


Prospective isolation of mesenchymal stem cells from multiple mammalian species using cross-reacting anti-human monoclonal antibodies.
Mesenchymal stem cells (MSCs) of human and nonhuman mammalian species are often studied for various applications in regenerative medicine research. These MSCs can be derived from human bone marrow (BM) and identified by their ability to form fibroblast-like colony forming units that develop into stromal like cells when expanded in culture. These cells are characterized by their spindle-shaped morphology, their characteristic phenotype (CD73(+), CD90(+), CD105(+), CD45⁻, and CD34⁻), and their ability to differentiate into cells of the osteogenic, adipogenic, and chondrogenic lineages. However, the identification and purification of MSCs from nonhuman mammalian species is hampered by the lack of suitable monoclonal antibodies (mAb). In this report, primary BM and cultured BM-derived MSCs of human and monkey, goat, sheep, dog, and pig were screened for cross-reactivity using a panel of 43 mAb, of which 22 react with either human BM mononuclear cells or cultured human MSCs. We found 7 mAb with specificity for CD271, MSCA-1 (W8B2 antigen), W4A5, CD56, W3C4 (CD349), W5C4, and 58B1, which showed interspecies cross-reactivity. These mAb proved to be useful for prospective sorting of MSCs from the BM of the 6 mammalian species studied as well as for the characterization of their cultured offspring. Flow sorting with the cross-reacting mAb resulted in up to 2400-fold enrichment of the clonogenic cell fraction (fibroblast-like colony forming units). This study provides an important contribution for the comparative prospective isolation of primary BM-MSCs and the characterization of cultured MSCs from multiple mammalian species for preclinical research.


BRAF pyrosequencing analysis aided by a lookup table.
BRAF mutations have substantial therapeutic, diagnostic, and prognostic significance, so detecting and specifying them is an important part of the workload of molecular pathology laboratories. Pyrosequencing assays are well suited for this analysis but can produce complex results. Therefore, we introduce a pyrosequencing lookup table based on Pyromaker that assists the user in generating hypotheses for solving complex pyrosequencing results. The lookup table contains all known mutations in the sequenced region and the positions in the dispensation sequence at which changes would occur with those mutations. We demonstrate the lookup table using a homebrew dispensation sequence for BRAF codons 596 to 605 as well as a commercially available kit-based dispensation sequence for codons 599 to 600. These results demonstrate that the homebrew dispensation sequence unambiguously identifies all known BRAF mutations in this region, whereas the kit-based dispensation sequence has one unresolvable degeneracy that could be solved with the addition of two injections. Using the lookup table and confirmatory virtual pyrogram, we unambiguously solved clinical pyrograms of the complex mutations V600K (c.1798_1799delGTinsAA), V600R (c.1798_1799delGTinsAG), V600D (c.1799_1800delTGinsAT), V600E (c.1799_1800delTGinsAA), and V600_K601delinsE (c.1799_1801delTGA). In addition, we used the approach to hypothesize and confirm a new mutation in human melanoma, V600_K601delinsEI (c.1799_1802delTGAAinsAAAT).


Small RNA and transcriptome deep sequencing proffers insight into floral gene regulation in Rosa cultivars.
Roses (Rosa sp.), which belong to the family Rosaceae, are the most economically important ornamental plants--making up 30% of the floriculture market. However, given high demand for roses, rose breeding programs are limited in molecular resources which can greatly enhance and speed breeding efforts. A better understanding of important genes that contribute to important floral development and desired phenotypes will lead to improved rose cultivars. For this study, we analyzed rose miRNAs and the rose flower transcriptome in order to generate a database to expound upon current knowledge regarding regulation of important floral characteristics. A rose genetic database will enable comprehensive analysis of gene expression and regulation via miRNA among different Rosa cultivars. We produced more than 0.5 million reads from expressed sequences, totalling more than 110 million bp. From these, we generated 35,657, 31,434, 34,725, and 39,722 flower unigenes from Rosa hybrid: 'Vital', 'Maroussia', and 'Sympathy' and Rosa rugosa Thunb., respectively. The unigenes were assigned functional annotations, domains, metabolic pathways, Gene Ontology (GO) terms, Plant Ontology (PO) terms, and MIPS Functional Catalogue (FunCat) terms. Rose flower transcripts were compared with genes from whole genome sequences of Rosaceae members (apple, strawberry, and peach) and grape. We also produced approximately 40 million small RNA reads from flower tissue for Rosa, representing 267 unique miRNA tags. Among identified miRNAs, 25 of them were novel and 242 of them were conserved miRNAs. Statistical analyses of miRNA profiles revealed both shared and species-specific miRNAs, which presumably effect flower development and phenotypes. In this study, we constructed a Rose miRNA and transcriptome database, and we analyzed the miRNAs and transcriptome generated from the flower tissues of four Rosa cultivars. The database provides a comprehensive genetic resource which can be used to better understand rose flower development and to identify candidate genes for important phenotypes.


Hunter-gatherers have less famine than agriculturalists.
The idea that hunter-gatherer societies experience more frequent famine than societies with other modes of subsistence is pervasive in the literature on human evolution. This idea underpins, for example, the 'thrifty genotype hypothesis'. This hypothesis proposes that our hunter-gatherer ancestors were adapted to frequent famines, and that these once adaptive 'thrifty genotypes' are now responsible for the current obesity epidemic. The suggestion that hunter-gatherers are more prone to famine also underlies the widespread assumption that these societies live in marginal habitats. Despite the ubiquity of references to 'feast and famine' in the literature describing our hunter-gatherer ancestors, it has rarely been tested whether hunter-gatherers suffer from more famine than other societies. Here, we analyse famine frequency and severity in a large cross-cultural database, in order to explore relationships between subsistence and famine risk. This is the first study to report that, if we control for habitat quality, hunter-gatherers actually had significantly less--not more--famine than other subsistence modes. This finding challenges some of the assumptions underlying for models of the evolution of the human diet, as well as our understanding of the recent epidemic of obesity and type 2 diabetes mellitus.


Environmental distribution and genetic diversity of vegetative compatibility groups determine biocontrol strategies to mitigate aflatoxin contamination of maize by Aspergillus flavus.
Maize infected by aflatoxin-producing Aspergillus flavus may become contaminated with aflatoxins, and as a result, threaten human health, food security and farmers' income in developing countries where maize is a staple. Environmental distribution and genetic diversity of A. flavus can influence the effectiveness of atoxigenic isolates in mitigating aflatoxin contamination. However, such information has not been used to facilitate selection and deployment of atoxigenic isolates. A total of 35 isolates of A. flavus isolated from maize samples collected from three agro-ecological zones of Nigeria were used in this study. Ecophysiological characteristics, distribution and genetic diversity of the isolates were determined to identify vegetative compatibility groups (VCGs). The generated data were used to inform selection and deployment of native atoxigenic isolates to mitigate aflatoxin contamination in maize. In co-inoculation with toxigenic isolates, atoxigenic isolates reduced aflatoxin contamination in grain by > 96%. A total of 25 VCGs were inferred from the collected isolates based on complementation tests involving nitrate non-utilizing (nit(-)) mutants. To determine genetic diversity and distribution of VCGs across agro-ecological zones, 832 nit(-) mutants from 52 locations in 11 administrative districts were paired with one self-complementary nitrate auxotroph tester-pair for each VCG. Atoxigenic VCGs accounted for 81.1% of the 153 positive complementations recorded. Genetic diversity of VCGs was highest in the derived savannah agro-ecological zone (H = 2.61) compared with the southern Guinea savannah (H = 1.90) and northern Guinea savannah (H = 0.94) zones. Genetic richness (H = 2.60) and evenness (E5 = 0.96) of VCGs were high across all agro-ecological zones. Ten VCGs (40%) had members restricted to the original location of isolation, whereas 15 VCGs (60%) had members located between the original source of isolation and a distance > 400 km away. The present study identified widely distributed VCGs in Nigeria such as AV0222, AV3279, AV3304 and AV16127, whose atoxigenic members can be deployed for a region-wide biocontrol of toxigenic isolates to reduce aflatoxin contamination in maize.


Flap Endonuclease 1 Limits Telomere Fragility on the Leading Strand.
The existence of redundant replication and repair systems that ensure genome stability underscores the importance of faithful DNA replication. Nowhere is this complexity more evident than in challenging DNA templates, including highly repetitive or transcribed sequences. Here, we demonstrate that flap endonuclease 1 (FEN1), a canonical lagging strand DNA replication protein, is required for normal, complete leading strand replication at telomeres. We find that the loss of FEN1 nuclease activity, but not DNA repair activities, results in leading strand-specific telomere fragility. Furthermore, we show that FEN1 depletion-induced telomere fragility is increased by RNA polymerase II inhibition and is rescued by ectopic RNase H1 expression. These data suggest that FEN1 limits leading strand-specific telomere fragility by processing RNA:DNA hybrid/flap intermediates that arise from co-directional collisions occurring between the replisome and RNA polymerase. Our data reveal the first molecular mechanism for leading strand-specific telomere fragility and the first known role for FEN1 in leading strand DNA replication. Because FEN1 mutations have been identified in human cancers, our findings raise the possibility that unresolved RNA:DNA hybrid structures contribute to the genomic instability associated with cancer.


Perivascular space in Parkinson's disease: Association with CSF amyloid/tau and cognitive decline.
Whether perivascular space (PVS) visible on magnetic resonance imaging (MRI) represents glymphatic dysfunction and whether this imaging marker is pathologic in Parkinson's disease (PD) have been controversial. The objective was to determine whether PVS visible on MRI is independently associated with cognitive decline in patients with PD, and to test whether pathologic proteins in the CSF (such as Aβ42) mediate the pathologic role of PVS. A total of 341 patients with Parkinson's disease from Parkinson's Progression Marker Initiative (PPMI) cohort was included in the present study. PVS in the basal ganglia (BG-PVS) and centrum semiovale were evaluated with a semiquantitative scale. Changes in the Montreal Cognitive Assessment (MoCA) score and the absolute MoCA score at the 3-year assessment were considered the main cognitive outcome. A multivariable linear regression model was used to test the association between PVS and cognitive decline. A mixed linear model and path analysis were used to test the interaction among PVS, CSF biomarkers and cognitive decline. BG-PVS was associated with cognitive decline in patients with PD at the 3-year follow-up independent of age, baseline cognition, motor and nonmotor function, presynaptic dopaminergic deficiency, and CSF biomarkers. The interaction between BG-PVS and Aβ42/tTau, Aβ42/pTau, and Aβ42 levels was significantly predictive of 3-year cognitive decline. Path analysis confirmed that CSF Aβ42/tTau levels partially mediated the pathologic effect of BG-PVS on cognitive outcome in PD. BG-PVS is independently associated with cognitive decline in PD, and this association may be partially mediated by toxic CSF proteins.


Engaging children through sport: examining the disconnect between program vision and implementation.
The purpose of this study was to provide recommendations to an organization trying to effectively implement nontraditional sport programming to reach a broader range of children and engage them in physical activity. This consultation-based qualitative study used data collected from 7 after-school sport program sites. Data were collected through participant observation and semistructured interviews with program instructors. The data were analyzed in 2 steps. First, descriptive coding was used to group observations and responses from each question, then pattern coding was used to find emerging themes. Researchers then compared both within and across program sites. Researchers found that enjoyment, ability, and language influenced interactions; age-appropriateness, engagement, and curriculum design impacted curriculum; and instructor roles and ongoing mentoring impacted effectiveness of training/support. A fundamental disconnect was evident between the program vision and the instructors' interpretation (and therefore, implementation) of the vision. Recommendations offered for practice include continued focus on curriculum design that can engage children at each level of development (grades K-5) and increased training and field support for instructors to ensure intended implementation of the programming.


The Linking Lives health education program: a randomized clinical trial of a parent-based tobacco use prevention program for african american and latino youths.
We evaluated the effectiveness of a parent-based add-on component to a school-based intervention to prevent cigarette smoking among African American and Latino middle school youths. Mother-adolescent dyads (n=1386) were randomly assigned to 2 groups: (1) a school-based smoking-prevention intervention or (2) the same intervention with a parent-based add-on component called Raising Smoke-Free Kids. Mothers in the experimental condition received the parent add-on component. Mothers in the control condition received information on selecting a high school. All adolescents received a version of Project Towards No Tobacco Use (TNT). The primary outcome was a reduction in adolescent cigarette smoking. Follow-up data were obtained from 1096 mother-adolescent dyads at 15 months postintervention. At follow-up, the odds of smoking cigarettes were reduced by 42% for adolescents in the parent add-on condition versus the TNT-only condition. Mothers in the parent add-on condition were more likely than were mothers in the TNT-only condition to set rules about risk-sensitive social activities and to be perceived as trustworthy by their child. Group differences also were found in the frequency and quality of mother-adolescent communication. Including parent add-on components in school-based smoking prevention programs can reduce smoking behavior on the part of inner-city middle school youths.


Treatment efficacy: hearing loss in children.
This article provides a review of the topic of treatment efficacy for children with hearing loss. Efficacy is related to a wide range of treatment goals in the areas of sensory and perceptual skill development, language development (regardless of communication modality), speech-production skill development, academic performance, and social-emotional growth. Topics addressed in this article include (a) the definition of hearing loss in children; (b) incidence and prevalence data; (c) the effects of childhood hearing loss on daily life, including language and literacy, speech perception and production, socialization and family dynamics; (d) the role of audiologists and speech-language pathologists in managing children with hearing loss; and (e) a summary of pertinent efficacy research for children with hearing loss. The analysis of the available research suggests that (a) early intervention for children who are deaf or hard of hearing has long-term positive effects on overall development; (b) a variety of communication modalities exist for this population, and research to date has been more descriptive than prognostic on the choice of modality; (c) sensory aids (hearing aids, tactile aids, and cochlear implants) provide different degrees of benefit for children in the areas of speech perception, production, and language development, depending upon the extent of their hearing loss; (d) few studies have addressed rates of learning and long-term outcomes, but existing data suggest that enriched programs provide some children with hearing loss with the ability to overcome developmental lags in language and academic skills.


Liver macrophages contribute to pancreatic cancer-related cachexia.
Cachexia is a devastating process especially in pancreatic cancer patients and contributes to their poor survival. We attempted to clarify the pathological and molecular changes that occur in the liver during the development of cachexia. Using immunohistochemistry we investigated the infiltration of inflammatory mononuclear cells in liver biopsies of pancreatic cancer patients with or without cachexia, and the potential relevance of the cells for the nutritional and inflammatory status. Additionally, these findings were compared with the patients' clinical parameters. We found a significantly higher amount of CD68 immunoreactive macrophages in liver cross sections of patients with pancreatic cancer and cachexia. The number of CD68-positive macrophages was significantly inversely correlated with the nutritional status. Additionally, in these CD68-positive areas a significant increase in IL-6 and IL-1 immunoreactive cells was localized. Moreover, we found significantly increased areas of CD68-positive macrophages in liver biopsies of patients with a more dedifferentiated (aggressive) grading of the tumor. In conclusion, these results suggest that a crucial interaction between the tumor, PBMCs, and the liver may play a central role in the development and regulation of cachexia. Furthermore, pancreatic cancer may be able to alter systemic organ function even without obvious metastatic disease.


Why Is It Just So Hard? Making Sense of End-of-Life Communication Between Adult Children and Their Terminally Ill Parental Figures.
The aging population in the United States is predicted to become one fifth of the population by 2050. With that increase, more individuals in the country will be experiencing chronic health conditions and the need for care, with end of life (EoL) becoming more of a topic that needs to be discussed. This study aims to explore the ways adult children talk to their parents about EoL, death, and dying. We discovered six themes: protection, meeting needs, guilt and regret, control, family dynamics, and communication type. Each of these was prevalent in responses to how adult children cope, how they cared for their parent, and how hope played into the interactions. We believe these themes will be helpful in developing a quantitative scale to study EoL further and be practical in helping adult children cope following death of their parent.


Crimean-Congo hemorrhagic fever: An emerging threat for the intensivist.
We present the case of a 55-year-old female, who presented with 15 days of fever with rash, pancytopenia, and altered behavior. She was investigated for routine causes of fever with rash and multi organ dysfunction and treated for the same. As she tested negative for all routine causes of such an illness and did not show improvement to therapy, she was investigated for Crimean-Congo hemorrhagic fever and tested positive for the same. She was started on ribavirin, but eventually succumbed to her illness. This disease has rarely been reported from the Northern India and we need to have high clinical suspicion for this deadly disease so that appropriate therapy can be started in time for the patient and prophylaxis given to all inadvertently exposed.


Analysis of sickness absence among employees of four NHS trusts.
To determine the value of using routinely collected sickness absence data as part of a health needs assessment of healthcare workers. Sickness absence records of almost 12900 NHS staff for one calendar year were analysed. Three measures of absence, the absence rate, the absence frequency rate, and the mean duration of absence, were assessed for the population and comparisons made between men and women, full and part time and different occupational groups of staff. Also, the main causes of sickness absence were found. Almost 60% of the study population had no spells of sickness absence in the year of study and almost 20% had only one spell of sickness absence. Female staff were more likely to have experienced sickness absence than male staff. Although absence due to conditions related to pregnancy were included in the analysis, the incidence of these was not sufficient to account for the higher rates of absence among female staff. In general, full time staff had greater rates of sickness absence than part time staff. 71% of all absences were of < 1 week duration. The main known causes of sickness absence were respiratory disorders, digestive disorders, and musculoskeletal disorders. The transition from units managed directly from the health board to trusts with individual responsibility for personnel issues at the time of data collection resulted in variations in the quality of data available for analysis. This together with the use of "dump" codes has influenced the quality of the analysis. However, such data should be available for analysis to tailor occupational health care to the needs of the population.


Long-read sequencing data analysis for yeasts.
Long-read sequencing technologies have become increasingly popular due to their strengths in resolving complex genomic regions. As a leading model organism with small genome size and great biotechnological importance, the budding yeast Saccharomyces cerevisiae has many isolates currently being sequenced with long reads. However, analyzing long-read sequencing data to produce high-quality genome assembly and annotation remains challenging. Here, we present a modular computational framework named long-read sequencing data analysis for yeasts (LRSDAY), the first one-stop solution that streamlines this process. Starting from the raw sequencing reads, LRSDAY can produce chromosome-level genome assembly and comprehensive genome annotation in a highly automated manner with minimal manual intervention, which is not possible using any alternative tool available to date. The annotated genomic features include centromeres, protein-coding genes, tRNAs, transposable elements (TEs), and telomere-associated elements. Although tailored for S. cerevisiae, we designed LRSDAY to be highly modular and customizable, making it adaptable to virtually any eukaryotic organism. When applying LRSDAY to an S. cerevisiae strain, it takes ∼41 h to generate a complete and well-annotated genome from ∼100× Pacific Biosciences (PacBio) running the basic workflow with four threads. Basic experience working within the Linux command-line environment is recommended for carrying out the analysis using LRSDAY.


An active protein kinase A (PKA) is involved in meiotic arrest of rat growing oocytes.
Reinitiation of meiosis in meiotically competent, fully grown mammalian oocytes is governed by a fall in intraoocyte cAMP concentrations and the subsequent inactivation of protein kinase A (PKA). A similar reduction in intraoocyte cAMP concentrations in growing, meiotically incompetent rat oocytes not leading to resumption of meiosis, questions the involvement of PKA in the regulation of meiosis at this early stage of oocyte development. We examined the possibility of whether PKA activity maintains growing oocytes in meiotic arrest and further explored the mode of activation of PKA under conditions of relatively low cAMP concentrations. Our experiment demonstrated that inactivation of PKA stimulates growing rat oocytes to resume meiosis, and elevates the activity of their maturation-promoting factor (MPF). We also found that the expressions of type I and type II regulatory subunits (RI and RII) of PKA are higher in growing and fully grown oocytes, respectively. In addition, we revealed that the common 1:1 ratio between the regulatory (R) and catalytic (C) subunits of PKA is apparently not abrogated and, in accordance PKA activity in growing oocyte-cell extract is fully dependent on cAMP. Finally, we identified in growing oocytes, the A kinase anchoring protein (AKAP) 140, which was previously depicted in fully grown oocytes. We conclude that an active PKA prevents growing oocytes from resuming meiosis. Our findings further suggest that relatively high abundance of the PKAI isoform and/or its subcellular compartmentalization, through interaction with AKAP140, could possibly account for the high basal PKA activity at relatively low intraoocyte cAMP concentrations.


Generalized Rosenfeld scalings for tracer diffusivities in not-so-simple fluids: mixtures and soft particles.
Rosenfeld [Phys. Rev. A 15, 2545 (1977)] originally noticed that casting the transport coefficients of simple monatomic equilibrium fluids in a specific dimensionless form makes them approximately single-valued functions of excess entropy. This observation has predictive value because, while the transport coefficients of dense fluids can be difficult to estimate from first principles, the excess entropy can often be accurately predicted from liquid-state theory. In this work, we use molecular simulations to investigate whether Rosenfeld's observation is a special case of a more general scaling law relating the tracer diffusivities of particles in mixtures to the excess entropy. Specifically, we study the tracer diffusivities, static structure, and thermodynamic properties of a variety of one- and two-component model fluid systems with either additive or nonadditive interactions of the hard-sphere or Gaussian-core form. The results of the simulations demonstrate that the effects of mixture concentration and composition, particle-size asymmetry and additivity, and strength of the interparticle interactions in these fluids are consistent with an empirical scaling law relating the excess entropy to a dimensionless (generalized Rosenfeld) form of tracer diffusivity, which we introduce here. The dimensionless form of the tracer diffusivity follows from knowledge of the intermolecular potential and the transport/thermodynamic behavior of fluids in the dilute limit. The generalized Rosenfeld scaling requires less information and provides more accurate predictions than either Enskog theory or scalings based on the pair-correlation contribution to the excess entropy. As we show, however, it also suffers from some limitations especially for systems that exhibit significant decoupling of individual component tracer diffusivities.


Commodity risk assessment of specified species of Lonicera potted plants from Turkey.
The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in the Commission Implementing Regulation (EU) 2018/2019 as 'High-risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by potted plants (2-4 years old) of specified Lonicera species produced in nurseries and that are imported from Turkey, taking into account the available scientific information, including the technical information provided by the NPPO of Turkey. The relevance of any pest for this Opinion was based on evidence following defined criteria listed in Section 4.1. Three species, the EU-quarantine pests Lopholeucaspis japonica and Meloidogyne chitwoodi and the protected zone quarantine pest Bemisia tabaci, fulfilled these criteria and were selected for further evaluation. For these pests, the risk mitigation measures proposed in the technical dossier from Turkey were evaluated taking into account the possible limiting factors. For these pests, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The estimated degree of pest freedom varies among the pests evaluated, with B. tabaci on evergreen species of Lonicera spp. being the pest most frequently expected on the imported plants. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9,293 and 10,000 plants per 10,000 would be free of B. tabaci.


Binding of pertechnetate to uranyl(VI) in aqueous solution. A density functional theory molecular dynamics study.
According to constrained Car-Parrinello molecular dynamics simulations and thermodynamic integration, the free binding energy between uranyl hydrate and pertechnetate in aqueous solution is significantly lower than that between uranyl and nitrate, namely, by 1.7 kcal mol(-1). This is the first study of the differential binding of these two ligands to uranyl, which can have implications for the separability of uranium and technetium during the reprocessing of nuclear waste.


Recruiting interns and residents to an osteopathic medical training program.
As more graduates of colleges of osteopathic medicine enroll in allopathic training programs and specialty programs, medical directors of osteopathic medical institutions are faced with the challenge of maintaining a well-trained housestaff. More important, perhaps, this exodus threatens the very core of osteopathic medicine, namely, its ability to produce well-trained primary care physicians. A recruitment plan used at the Chicago College of Osteopathic Medicine and its affiliates is presented here. The plan emphasizes the importance of trainee involvement in their own education as well as the use of residents in recruiting interns and specialty residents.


Heritability of blood pressure and hemodynamics in African- and European-American youth.
Hypertension prevalence is much higher in African-Americans (AAs) than in European-Americans (EAs). It is unknown whether this difference is related to potential ethnic differences in the relative contribution of genes and environment to population variation in blood pressure and underlying hemodynamics. We studied 308 EA and 226 AA twin pairs, including monozygotic and dizygotic twins, of the same as well as the opposite sex (mean+/-SD age, 14.7+/-3.1 years). Supine resting systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure, and heart rate (HR) were measured by a Dinamap instrument and hemodynamics (stroke volume, cardiac index, and total peripheral resistance [TPR] index) by impedance cardiography. Ethnic and sex effects on genetic and environmental contributions to resting blood pressure and hemodynamics were estimated by genetic model fitting. For most measures, the best-fitting model showed no differences in heritability between AAs and EAs or between males and females, with heritabilities of 0.50 for cardiac index, of 0.64 for HR, and of SBP, pulse pressure, and stroke volume in between. Heritability of DBP was 0.45 in EAs and 0.58 in AAs with no effect of sex. For TPR index in EAs, 46% of the variance could be attributed to familial effects, but no significant distinction could be made between shared environmental and genetic factors. Heritability of TPR index in AAs was 0.51. Adjustment for obesity yielded virtually identical heritabilities. In summary, relative influences of genetic and environmental factors on blood pressure and hemodynamics in AA and EA youth are similar and independent of (genes for) obesity.


Reconstruction and analysis of the lncRNA-miRNA-mRNA network based on competitive endogenous RNA reveal functional lncRNAs in rheumatoid arthritis.
Rheumatoid arthritis (RA) is an autoimmune disease with an unknown etiology, occurring in approximately 1.0% of general population. More and more studies have suggested that long non-coding RNAs (lncRNAs) could play important roles in various biological processes and be associated with the pathogenesis of different kinds of diseases including RA. Although a large number of lncRNAs have been found, our knowledge of their function and physiological/pathological significance is still in its infancy. In order to reveal functional lncRNAs and identify the key lncRNAs in RA, we reconstructed a global triple network based on the competitive endogenous RNA (ceRNA) theory using the data from National Center for Biotechnology Information Gene Expression Omnibus and our previous paper. Meanwhile, Gene Ontology (GO) and pathway analysis were performed using Cytoscape plug-in BinGO and Database for Annotation, Visualization, and Integration Discovery (DAVID), respectively. We found that the lncRNA-miRNA-mRNA network was composed of 7 lncRNA nodes, 90 mRNA nodes, 24 miRNA nodes, and 301 edges. The functional assay showed that 147 GO terms and 23 pathways were enriched. In addition, three lncRNAs (S5645.1, XR_006437.1, J01878) were highly related to RA, and therefore, were selected as key lncRNAs. This study suggests that specific lncRNAs are associated with the development of RA, and three lncRNAs (S5645.1, XR_006437.1, J01878) could be used as potential diagnostic biomarkers and therapeutic targets.


Non-destructive characterisation of mesenchymal stem cell differentiation using LC-MS-based metabolite footprinting.
Bone regeneration is a complex biological process where major cellular changes take place to support the osteogenic differentiation of mesenchymal bone progenitors. To characterise these biological changes and better understand the pathways regulating the formation of mature bone cells, the metabolic profile of mesenchymal stem cell (MSC) differentiation in vitro has been assessed non-invasively during osteogenic (OS) treatment using a footprinting technique. Liquid chromatography (LC)-mass spectrometry (MS)-based metabolite profiling of the culture medium was carried out in parallel to mineral deposition and alkaline phosphatase activity which are two hallmarks of osteogenesis in vitro. Metabolic profiles of spent culture media with a combination of univariate and multivariate analyses investigated concentration changes of extracellular metabolites and nutrients linked to the presence of MSCs in culture media. This non-invasive LC-MS-based analytical approach revealed significant metabolic changes between the media from control and OS-treated cells showing distinct effects of MSC differentiation on the environmental footprint of the cells in different conditions (control vs. OS treatment). A subset of compounds was directly linked to the osteogenic time-course of differentiation, and represent interesting metabolite candidates as non-invasive biomarkers for characterising the differentiation of MSCs in a culture medium.


Exposure of children to phthalates and the impact of consumer practices in Slovakia.
Phthalates are environmental pollutants that can enter the human body by ingestion, inhalation and dermal absorption. Food constitutes the most important source of human exposure to these chemicals. The aim of our study was the biological monitoring of exposure to eight phthalate metabolites in children (n=107), 10-12 years of age, living in eastern Slovakia. Additionally, we analysed some associations between anthropometric measures, questionnaire data (i.e. eating and drinking habits, practice of personal care products) and concentrations of phthalate metabolites. Because of the short half-life of phthalates, within 24-48 h, we used 24-h recalls to assess dietary intakes. We used high-performance liquid chromatography and tandem mass spectrometry for the analysis of spot urine samples to determine concentrations of phthalate metabolites mono-ethyl phthalate (MEP), mono-n-butyl phthalate, mono-iso-butyl phthalate, mono-benzyl phthalate (MBzP), mono (2-ethyl-5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl) phthalate (5oxo-MEHP), mono-carboxy pentyl and mono (2-ethylhexyl) phthalate (MEHP). We found statistically significant association between consumer practices and concentration of some phthalate metabolites, concretely consumption of milk and dairy products with MBzP and margarine with MEP (p<0.01 in both cases) and margarine with 5oxo-MEHP, hot beverages with 5OH-MEHP, baguettes and semifinished products with MEP (p<0.05 in all cases). Further, we found relationship between use of cosmetic products and phthalate concentrations, nail polish application and MEP and use of body lotion and MEHP (p<0.05 in both cases). We concluded that consumer practices (including eating and drinking habits and personal care) represent the substantial source of phthalate exposure in Slovak children.


Vertex models: from cell mechanics to tissue morphogenesis.
Tissue morphogenesis requires the collective, coordinated motion and deformation of a large number of cells. Vertex model simulations for tissue mechanics have been developed to bridge the scales between force generation at the cellular level and tissue deformation and flows. We review here various formulations of vertex models that have been proposed for describing tissues in two and three dimensions. We discuss a generic formulation using a virtual work differential, and we review applications of vertex models to biological morphogenetic processes. We also highlight recent efforts to obtain continuum theories of tissue mechanics, which are effective, coarse-grained descriptions of vertex models.This article is part of the themed issue 'Systems morphodynamics: understanding the development of tissue hardware'.


Nitrogen-fixing bacteria in Eucalyptus globulus plantations.
Eucalypt cultivation is an important economic activity worldwide. In Portugal, Eucalyptus globulus plantations account for one-third of the total forested area. The nutritional requirements of this crop have been well studied, and nitrogen (N) is one of the most important elements required for vegetal growth. N dynamics in soils are influenced by microorganisms, such as diazotrophic bacteria (DB) that are responsible for biological nitrogen fixation (BNF), so the aim of this study was to evaluate and identity the main groups of DB in E. globulus plantations. Samples of soil and root systems were collected in winter and summer from three different Portuguese regions (Penafiel, Gavião and Odemira). We observed that DB communities were affected by season, N fertilization and moisture. Furthermore Bradyrhizobium and Burkholderia were the most prevalent genera in these three regions. This is the first study describing the dynamic of these bacteria in E. globulus plantations, and these data will likely contribute to a better understanding of the nutritional requirements of eucalypt cultivation and associated organic matter turnover.


Serenoa repens for benign prostatic hyperplasia.
Benign prostatic hyperplasia (BPH) is a nonmalignant enlargement of the prostate, which can lead to obstructive and irritative lower urinary tract symptoms (LUTS). The pharmacologic use of plants and herbs (phytotherapy) for the treatment of LUTS associated with BPH is common. The extract of the berry of the American saw palmetto, or dwarf palm plant, Serenoa repens (SR), which is also known by its botanical name of Sabal serrulatum, is one of several phytotherapeutic agents available for the treatment of BPH. This systematic review aimed to assess the effects and harms of Serenoa repens in the treatment of men with LUTS consistent with BPH. We searched for trials in general and in specialized databases, including the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE®, EMBASE, CINAHL®, Web of Science, SCOPUS, BIOSIS Previews®, LILACS, ClinicalTrials.gov, Controlled-Trials.com, World Health Organization (WHO), and Google Scholar. We also handsearched systematic reviews, references, and clinical practice guidelines. There were no language restrictions. Trials were eligible if they randomized men with symptomatic BPH to receive preparations of SR (alone or in combination) for at least four weeks in comparison with placebo or other interventions, and included clinical outcomes, such as urologic symptom scales, symptoms, and urodynamic measurements. Eligibility was assessed by at least two independent observers (JT, RM). One review author (JT) extracted Information on patients, interventions, and outcomes which was then checked by another review author (RM). The main outcome measure for comparing the effectiveness of SR with active or inert controls was change in urologic symptom-scale scores, with validated scores taking precedence over non validated ones. Secondary outcomes included changes in nocturia and urodynamic measures. The main outcome measure for harms was the number of men reporting side effects. In a meta-analysis of two high quality long-term trials (n = 582), Serenoa repens therapy was not superior to placebo in reducing LUTS based on the AUA (mean difference (MD) 0.25 points, 95% confidence interval (CI) -0.58 to 1.07). A 72 week trial with high quality evidence, using the American Urological Association Symptom Score Index, reported that SR was not superior to placebo at double and triple doses. In the same trial the proportions of clinical responders (≥ three-point improvement) were nearly identical (42.6% and 44.2% for SR and placebo, respectively), and not significant (RR 0.96, 95% CI 0.76 to 1.22).This update, which did not change our previous conclusions, included two new trials with 444 additional men, an 8.5% (5666/5222) increase from our 2009 updated review, and a 28.8% (1988/1544) increase for our main comparison, SR monotherapy versus placebo control (17 trials). Overall, 5666 men were assessed from 32 randomized, controlled trials, with trial lengths from four to 72 weeks. Twenty-seven trials were double blinded and treatment allocation concealment was adequate in 14.In a trial of high quality evidence (N = 369), versus placebo, SR did not significantly decrease nightly urination on the AUA Nocturia scale (range zero to five) at 72 weeks follow-up (one-sided P = 0.19).The three high quality, moderate-to-long term trials found peak urine flow was not improved with Serenoa repens compared with placebo (MD 0.40 mL/s, 95% CI -0.30 to 1.09).Comparing prostate size (mean change from baseline), one high quality 12-month trial (N = 225) reported no significant difference between SR and placebo (MD -1.22 cc, 95% CI -3.91 to 1.47). Serenoa repens, at double and triple doses, did not improve urinary flow measures or prostate size in men with lower urinary tract symptoms consistent with BPH.


Protease-Activated Receptor 2 Facilitates Bacterial Dissemination in Pneumococcal Pneumonia.
Streptococcus pneumoniae is the most common causative pathogen in community-acquired pneumonia. Protease-activated receptor 2 (PAR2) is expressed by different cell types in the lungs and can mediate inflammatory responses. We sought to determine the role of PAR2 during pneumococcal pneumonia. Pneumococcal pneumonia or sepsis was induced in wild-type and PAR2 knock-out (Par2-/-) mice by infection with viable S. pneumoniae. Par2-/- mice demonstrated improved host defense, a largely preserved lung barrier integrity, and reduced mortality during pneumococcal pneumonia. PAR2 deficiency did not influence bacterial growth after intravenous infection. Inhibition of the endogenous PAR2 activating proteases tissue factor/factor VIIa or tryptase did not impact on bacterial burdens during pneumonia. In a PAR2 reporter cell line it was demonstrated that S. pneumoniae-derived proteases are able to cleave PAR2. These results show that S. pneumoniae is able to cleave and exploit PAR2 to disseminate systemically from the airways.


Early induction of apoptosis in hematopoietic cell lines after exposure to flavopiridol.
Flavopiridol (NSC 649890; Behringwerke L86-8275, Marburg, Germany), is a potent inhibitor of cyclin dependent kinases (CDKs) 1, 2, and 4. It has potent antiproliferative effects in vitro and is active in tumor models in vivo. While surveying the effect of flavopiridol on cell cycle progression in different cell types, we discovered that hematopoietic cell lines, including SUDHL4, SUDHL6 (B-cell lines), Jurkat, and MOLT4 (T-cell lines), and HL60 (myeloid), displayed notable sensitivity to flavopiridol-induced apoptosis. For example, after 100 nmol/L for 12 hours, SUDHL4 cells displayed a similar degree of DNA fragmentation to that shown by the apoptosis-resistant PC3 prostate carcinoma cells only after 3,000 nmol/L for 48 hours. After exposure to 1,000 nmol/L flavopiridol for 12 hours, typical apoptotic morphology was observed in SUDHL4 cells, but not in PC3 prostate carcinoma cells despite comparable potency (SUDHL4: 120 nmol/L; PC3: 203 nmol/L) in causing growth inhibition by 50% (IC50). Flavopiridol did not induce topoisomerase I or II cleavable complex activity. A relation of p53, bcl2, or bax protein levels to apoptosis in SUDHL4 was not appreciated. While flavopiridol caused cell cycle arrest with decline in CDK1 activity in PC3 cells, apoptosis of SUDHL4 cells occurred without evidence of cell cycle arrest. These results suggest that antiproliferative activity of flavopiridol (manifest by cell cycle arrest) may be separated in different cell types from a capacity to induce apoptosis. Cells from hematopoietic neoplasms appear in this limited sample to be very susceptible to flavopiridol-induced apoptosis and therefore clinical trials in hematopoietic neoplasms should be of high priority.


Modelling malaria population structure and its implications for control.
Mathematical models of malaria transmission have been used to inform the design of malaria control programs since the mid 20th century, and many of these models have provided useful insights into the complexity of the disease. Among developing countries, however and particularly in sub-Saharan Africa, malaria remains a major cause of morbidity and mortality. One of the main difficulties in controlling the most virulent human malaria parasite, Plasmodium falciparum, is its genetic diversity, which confounds attempts to design an effective vaccine. The population structure of P. falciparum remains poorly understood but plays a key role in determining epidemiological patterns of disease and the development of immunity. We discuss the seminal model of malaria transmission developed by Ross and MacDonald, and the modifications that have been made since to include more realism. We show that age profiles of disease and serological data support a theoretical model in which the parasite population is diverse and structured into several antigenic types and highlight the implications of this structure for controlling malaria. Lastly, we discuss the current sequence data on parasite antigen genes that are important for the aquisition of immunity, and the results of a new analysis of P. falciparum population structure at the genomic level.


Palliative Endoscopic Ultrasound Biliary Drainage for Advanced Malignant Biliary Obstruction: Should It Replace the Percutaneous Approach?
Endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic biliary drainage (PTBD) are the standard of care in malignant biliary obstruction cases. Recently, endoscopic ultrasound-guided biliary drainage (EUS-BD) has been widely used after unsuccessful ERCP. However, the patient's clinical impact of EUS-BD over PTBD is still not obvious. Therefore, this case series study aims to evaluate the clinical outcomes of patients with advanced malignant biliary obstruction who underwent EUS-BD after failed ERCP. A retrospective database study was performed between January 2016 and June 2018 in patients with advanced malignant biliary obstruction. Patients were consecutively enrolled without randomization. Treatment options consisted of ERCP and PTBD or EUS-BD if ERCP failed. Based on 144 biliary obstruction cases, 38 patients were enrolled; 24 (63.2%) were men. The patients' mean age was 66.8 ± 12.36 years. The most common cause of malignant biliary obstruction was pancreatic cancer (44.7%). Biliary drainage was achieved by ERCP (39.5%), PTBD (39.5%), and EUS-BD (21.1%). The technical success rate was 86.7% by PTBD and 87.5% by EUS-BD (p = 1.000), while the clinical success rate was 93.3% by PTBD and 62.5% by EUS-BD (p = 0.500). The median survival in patients who underwent PTBD versus those wo underwent EUS-BD was 11 versus 3 months (log-rank p = 0.455). In conclusion, there is no significant advantage of EUS-BD when compared to PTBD in terms of clinical success and survival benefit in advanced malignant biliary obstruction.


Sequencing of the core MHC region of black grouse (Tetrao tetrix) and comparative genomics of the galliform MHC.
The MHC, which is regarded as the most polymorphic region in the genomes of jawed vertebrates, plays a central role in the immune system by encoding various proteins involved in the immune response. The chicken MHC-B genomic region has a highly streamlined gene content compared to mammalian MHCs. Its core region includes genes encoding Class I and Class IIB molecules but is only ~92Kb in length. Sequences of other galliform MHCs show varying degrees of similarity as that of chicken. The black grouse (Tetrao tetrix) is a wild galliform bird species which is an important model in conservation genetics and ecology. We sequenced the black grouse core MHC-B region and combined this with available data from related species (chicken, turkey, gold pheasant and quail) to perform a comparative genomics study of the galliform MHC. This kind of analysis has previously been severely hampered by the lack of genomic information on avian MHC regions, and the galliformes is still the only bird lineage where such a comparison is possible. In this study, we present the complete genomic sequence of the MHC-B locus of black grouse, which is 88,390 bp long and contains 19 genes. It shows the same simplicity as, and almost perfect synteny with, the corresponding genomic region of chicken. We also use 454-transcriptome sequencing to verify expression in 17 of the black grouse MHC-B genes. Multiple sequence inversions of the TAPBP gene and TAP1-TAP2 gene block identify the recombination breakpoints near the BF and BLB genes. Some of the genes in the galliform MHC-B region also seem to have been affected by selective forces, as inferred from deviating phylogenetic signals and elevated rates of non-synonymous nucleotide substitutions. We conclude that there is large synteny between the MHC-B region of the black grouse and that of other galliform birds, but that some duplications and rearrangements have occurred within this lineage. The MHC-B sequence reported here will provide a valuable resource for future studies on the evolution of the avian MHC genes and on links between immunogenetics and ecology of black grouse.


Avian migrants adjust migration in response to environmental conditions en route.
The onset of migration in birds is assumed to be primarily under endogenous control in long-distance migrants. Recently, climate changes appear to have been driving a rapid change in breeding area arrival. However, little is known about the climatic factors affecting migratory birds during the migration cycle, or whether recently reported phenological changes are caused by plastic behavioural responses or evolutionary change. Here, we investigate how environmental conditions in the wintering areas as well as en route towards breeding areas affect timing of migration. Using data from 1984 to 2004 covering the entire migration period every year from observatories located in the Middle East and northern Europe, we show that passage of the Sahara Desert is delayed and correlated with improved conditions in the wintering areas. By contrast, migrants travel more rapidly through Europe, and adjust their breeding area arrival time in response to improved environmental conditions en route. Previous studies have reported opposing results from a different migration route through the Mediterranean region (Italy). We argue that the simplest explanation for different phenological patterns at different latitudes and between migratory routes appears to be phenotypic responses to spatial variability in conditions en route.


Peripheral Blood Leukocyte Production of BDNF following Mitogen Stimulation in Early Onset and Regressive Autism.
Brain-derived neurotrophic factor (BDNF) is critical for neuronal differentiation and synaptic development. BDNF is also implicated in the development of psychological disorders including depression, bipolar disorder and schizophrenia. Previously, elevated BDNF levels were observed in neonatal blood samples from infants who were later diagnosed with autism when compared with children who developed normally, suggesting that BDNF may be involved in the development of autism. BDNF is produced by activated brain microglial cells, a cellular phenotype that shares several features with peripheral macrophages, suggesting an important role for the immune system in BDNF production. We hypothesized that under mitogenic stimulation, peripheral blood mononuclear cells obtained from children with autism may have altered BDNF production compared with age-matched typically developing control subjects. In addition, we examined the differences between the production of BDNF in classic/early-onset autism and children who had a regressive form of autism. We show here that plasma levels of BDNF levels are increased in children with autism, especially in early onset autism subjects. Furthermore, under mitogenic stimulation with PHA and LPS, BDNF production is significantly increased in children with autism compared with typically developing subjects. However, stimulation with tetanus toxoid results in a decreased response in children with autism. This data suggest that immune cell-derived production of BDNF could be an important source for the increased BDNF that is detected in some subjects with autism. As a neurotrophic factor produced by immune cells, BDNF could help elucidate the role of the immune system in neurodevelopment and neuronal maintenance, which may be dysregulated in autism.


Place-Based Case Studies: A New Approach to an Effective Teaching Practice.
Case-based approaches have been used extensively in STEM classrooms to enhance the real-world applicability of course content. Prior research in the bioeducation field indicates, specifically, that such methods lead to increases in students' conceptual understanding and affect in the discipline relative to more traditional methods. Despite these outcomes, the majority of case study exercises are formatted in a generalist manner. In other words, the content and context of the case study itself are not framed around the communities in which the students live. In an effort to address this concern, we developed and implemented a series of place-based case study (PBCS) exercises within the introductory cell and molecular biology courses at our institutions. A comparative, quasi-experimental approach was used to evaluate the impact of PBCSs versus non-PBCSs on cognitive and non-cognitive student outcomes. Results indicated that both PBCSs and non-PBCSs led to increases in students' content knowledge; however, no statistically significant difference existed in post-exercise performance between the PBCS and non-PBCS cohorts at the University of Texas, for instance, after controlling for confounding factors. Importantly, data also revealed that students within the PBCS cohort agreed more strongly that the case studies provided them with a better understanding of how scientific advancements and research impacted the community in which they lived than did their peers in the non-PBCS cohort. Collectively, these outcomes suggest that PBCSs offer a scalable, classroom-based approach to engage students in relevant, practical experiences that are of direct interest to them and, ideally, the broader scientific community.


Case 01-2017 - Primary vitreoretinal lymphoma (PVRL): report of a case and update of literature from 1942 to 2016.
Primary vitreoretinal lymphoma (PVRL), as a subset of primary central nervous system lymphoma (PCNSL), is a rare and fatal ocular malignancy. Most PVRL masquerades as chronic posterior uveitis, which makes the clinical diagnosis challenging. Vitreous cells, subretinal lesions and imaging techniques are essential for clinical diagnosis. Importantly, cytopathology/histopathology identification of malignant cells is the gold standard for the diagnosis of PVRL. In addition, molecular detection of immunoglobulin heavy chain (IgH) or T cell receptor (TCR) gene rearrangements, immunophenotyping for cell markers, and cytokine analysis of interleukine-10 elevation are often used as adjunct procedures. Current management of PVRL involves local radiation, intravitreal chemotherapy (methotrexate and rituximab), with or without systemic chemotherapy depending on the involvement of non-ocular tissues. In cases with concomitant PCNSL, systemic high-dose methotrexate/rituximab based therapy in conjunction with local therapy, whole brain radiotherapy and/or autologous stem cell transplantation is considered. Although PVRL normally responds well to initial treatment, high rates of relapse and CNS involvement usually lead to poor prognosis and limited survival. A professional team of medical experts in ophthalmologists, ocular pathologists, neuro-oncologists and hemato-oncologists is essential for optimizing patient management.


Insect sex-pheromone signals mediated by specific combinations of olfactory receptors.
We describe two male-specific olfactory receptors (ORs) in the silk moth, Bombyx mori, that are mutually exclusively expressed in a pair of adjacent pheromone-sensitive neurons of male antennae: One is specifically tuned to bombykol, the sex pheromone, and the other to bombykal, its oxidized form. Both pheromone ORs are coexpressed with an OR from the highly conserved insect OR subfamily. This coexpression promotes the functional expression of pheromone receptors and confers ligand-stimulated nonselective cation channel activity. The same effects were also observed for general ORs. Both odorant and pheromone signaling pathways are mediated by means of a common mechanism in insects.


Effects of panduratin A isolated from Kaempferia pandurata ROXB. on the expression of matrix metalloproteinase-9 by porphyromonas gingivalis supernatant-induced KB cells.
Periodontitis is initiated by accumulation of microbial plaque and activation of gingival inflammation through overexpression of matrix metalloproteinases (MMPs), leading to tissue destruction. Natural MMP inhibitors may be developed as therapeutic agents against periodontitis. In this study, panduratin A, a natural bioactive compound isolated from Kaempferia pandurata ROXB., was used to test its in vitro inhibitory activity against MMP-9 secretion from Porphyromonas gingivalis supernatant-induced human oral epidermoid carcinoma KB cells. Gelatin zymography, Western blot and RT-PCR analyses were performed to evaluate MMP-9 expression. The gelatin zymograms revealed that the main gelatinase secreted by P. gingivalis supernatant-induced KB cells migrated at 92 kDa, representing MMP-9. MMP-9 protein and mRNA levels were significantly decreased after panduratin A treatment (p<0.05). In contrast, panduratin A had no effect on tissue inhibitor of metalloproteinases (TIMP)-1 and TIMP-2 mRNA. Panduratin A also suppressed urokinase type plasminogen activator (uPA) mRNA expression. These results suggest that panduratin A could potentially prevent periodontal inflammation by decreasing the levels of MMP-9 protein and mRNA.


Lunar-modulated geomagnetic orientation by a marine mollusk.
Behavioral experiments indicated that the marine opisthobranch mollusk Tritonia diomedea can derive directional cues from the magnetic field of the earth. The magnetic direction toward which nudibranchs spontaneously oriented in the geomagnetic field showed recurring patterns of variation correlated with lunar phase, suggesting that the behavioral response to magnetism is modulated by a circa-lunar rhythm. The discovery of a magnetic sense in a mollusk with giant, reidentifiable neurons provides a unique opportunity to study the cellular mechanisms underlying magnetic field detection.


Meta-Analysis of Efficacy of Mirtazapine as an Adjunctive Treatment of Negative Symptoms in Schizophrenia.
Despite advances made in treating the positive symptoms of schizophrenia, treatment of negative symptoms remains an unmet therapeutic need. Adjunctive mirtazapine has shown promise for treatment of negative symptoms in several small clinical trials. To assess the efficacy of mirtazapine as an adjunctive treatment of negative symptoms in patients with chronic schizophrenia via meta-analysis. A systematic literature review of articles in English and Spanish was conducted in November 2011 by searching PubMed, the Cochrane Library, the Clinical Trial Registry of the NIH, and SIGLE (System for Grey Literature in Europe). Free text search terms for PubMed were "schizophrenia," "negative symptoms" and "mirtazapine." Publication date was not a limitation. Studies of people with schizophrenia/schizoaffective disorder were included in the meta-analysis if they were randomized, double-blind, and used the Positive and Negative Syndrome Scale (PANSS) as an outcome measure. Nine studies were initially identified. Five studies were included in the meta-analysis; 1 study was excluded for not using the PANSS, 3 were excluded as representing duplicate publications and open-label phases of one of the selected randomized control trials. Studies varied in the quality of their selection for participants with primary negative symptoms. Three of the 5 studies showed significant improvement in negative symptoms individually. The overall analysis showed improvement in negative symptoms with an effect size of 1.00 (0.084-1.918), which was statistically significant (p=0.032). Data from the negative symptoms subscale of the PANSS from 169 subjects was used in a forest plot to illustrate the relative strength of treatment effects. The variation in standard median deviation (SMD) attributable to heterogeneity was 27.35 %, indicating a high degree of heterogeneity. This meta-analysis supports the hypothesis that adding mirtazapine to treatment with antipsychotics can improve negative symptoms in schizophrenia. However, additional studies with more stringent negative symptom selection criteria and homogeneous use of antipsychotics are needed.


Peer victimization during middle childhood as a lead indicator of internalizing problems and diagnostic outcomes in late adolescence.
We examined evidence that peer victimization in middle childhood is a lead indicator of internalizing behavior problems and diagnostic outcomes during adolescence. This research was conducted as part of an ongoing multisite longitudinal investigation. The participants were 388 children (198 boys, 190 girls). Peer victimization was assessed with a peer nomination inventory that was administered when the average age of the participants was approximately 8.5 years. Internalizing problems were assessed using a behavior problem checklist completed by mothers in 9 consecutive years, and a structured clinical interview was administered to the participants in the summer following high school graduation (10-11 years after the victimization assessment). Peer victimization in middle childhood was correlated with internalizing problems on a bivariate basis through the late years of adolescence. Multilevel analyses also revealed associations between peer victimization and increases in internalizing problems over time. In addition, peer victimization had a modest link to unipolar depressive disorders in late adolescence. Victimization in the peer group during middle childhood appears to be a marker of long-term risk for internalizing behavior problems and unipolar depression.


Hypocretin/Orexin regulation of dopamine signaling and cocaine self-administration is mediated predominantly by hypocretin receptor 1.
Extensive evidence suggests that the hypocretins/orexins influence cocaine reinforcement and dopamine signaling via actions at hypocretin receptor 1. By comparison, the involvement of hypocretin receptor 2 in reward and reinforcement processes has received relatively little attention. Thus, although there is some evidence that hypocretin receptor 2 regulates intake of some drugs of abuse, it is currently unclear to what extent hypocretin receptor 2 participates in the regulation of dopamine signaling or cocaine self-administration, particularly under high effort conditions. To address this, we examined the effects of hypocretin receptor 1, and/or hypocretin receptor 2 blockade on dopamine signaling and cocaine reinforcement. We used in vivo fast scan cyclic voltammetry to test the effects of hypocretin antagonists on dopamine signaling in the nucleus accumbens core and a progressive ratio schedule to examine the effects of these antagonists on cocaine self-administration. Results demonstrate that blockade of either hypocretin receptor 1 or both hypocretin receptor 1 and 2 significantly reduces the effects of cocaine on dopamine signaling and decreases the motivation to take cocaine. In contrast, blockade of hypocretin receptor 2 alone had no significant effects on dopamine signaling or self-administration. These findings suggest a differential involvement of the two hypocretin receptors, with hypocretin receptor 1 appearing to be more involved than hypocretin receptor 2 in the regulation of dopamine signaling and cocaine self-administration. When considered with the existing literature, these data support the hypothesis that hypocretins exert a permissive influence on dopamine signaling and motivated behavior via preferential actions on hypocretin receptor 1.


Etiology and Management of Allergic Eyelid Dermatitis.
Allergic dermatitis is a common but often misdiagnosed condition that can present with a variety of findings including inflammation, eyelid malposition, and tearing. This study was performed to identify likely etiologies, along with presenting findings and treatment strategies for allergic dermatitis. A retrospective review of clinical records was performed of patients diagnosed with allergic dermatitis in academic oculoplastics practices from 2002 to 2014. Initial consultation included review of medications and cosmeceuticals. Suspected allergens were discontinued and an ophthalmic steroid was applied. Persistent allergic dermatitis led to further medication changes or formal allergy testing in an effort to identify the causative agent. Sixty-one patients were identified; average age was 66 years old (range: 33-94), and 45 of the patients were women (74%). Average follow up was 7 months (range: 1-60 months). Reasons for referral included epiphora (31.2%), ectropion (24.6%), blepharitis/dermatitis (18.0%), and "droopy" eyelid (14.8%). Presenting symptoms included irritation (77.1%) and tearing (50.8%). Average duration of symptoms was 16.5 months (range: 2 days-8 years). The 3 most common etiologies were eye drops (54.2%), creams/lotions (24.6%), and cosmetics (13.1%). Rubbing/manipulation was also thought to be a significant factor in 30% of the patients. Overall, the initial treatment regimen led to at least partial resolution in 88% of patients and 66% experienced complete resolution. Patients resistant to therapy were referred for patch testing. At follow up, 98% of patients had improvement in their symptoms and 90% of patients had complete resolution of their symptoms. Allergic eyelid dermatitis is commonly related to eye drops and topical cosmetics or skin care products. Identification and elimination of causative agents is the mainstay of management. Topical corticosteroids often facilitate resolution of the associated inflammation. Therapy resistant patients may benefit from formal allergy testing.


Species invasion shifts the importance of predator dependence.
The strength of interference between foraging individuals can influence per capita consumption rates, with important consequences for predator and prey populations and system stability. Here we demonstrate how the replacement of a previously established invader, the predatory crab Carcinus maenas, by the recently invading predatory crab Hemigrapsus sanguineus shifts predation from a species that experiences strong predator interference (strong predator dependence) to one that experiences weak predator interference (weak predator dependence). We demonstrate using field experiments that differences in the strength of predator dependence persist for these species both when they forage on a single focal prey species only (the mussel Mytilus edulis) and when they forage more broadly across the entire prey community. This shift in predator dependence with species replacement may be altering the biomass across trophic levels, consistent with theoretical predictions, as we show that H. sanguineus populations are much larger than C. maenas populations throughout their invaded ranges. Our study highlights that predator dependence may differ among predator species and demonstrates that different predatory impacts of two conspicuous invasive predators may be explained at least in part by different strengths of predator dependence.


The bromodomain inhibitor JQ1+ reduces calcium-sensing receptor activity in pituitary cell lines.
Corticotrophinomas represent 10% of all surgically removed pituitary adenomas, however, current treatment options are often not effective, and there is a need for improved pharmacological treatments. Recently, JQ1+, a bromodomain inhibitor that promotes gene transcription by binding acetylated histone residues and recruiting transcriptional machinery, has been shown to reduce proliferation in a murine corticotroph cell line, AtT20. RNA-Seq analysis of AtT20 cells following treatment with JQ1+ identified the calcium-sensing receptor (CaSR) gene as significantly downregulated, which was subsequently confirmed using real-time PCR and Western blot analysis. CaSR is a G protein-coupled receptor that plays a central role in calcium homeostasis but can elicit non-calcitropic effects in multiple tissues, including the anterior pituitary where it helps regulate hormone secretion. However, in AtT20 cells, CaSR activates a tumour-specific cAMP pathway that promotes ACTH and PTHrP hypersecretion. We hypothesised that the Casr promoter may harbour binding sites for BET proteins, and using chromatin immunoprecipitation (ChIP)-sequencing demonstrated that the BET protein Brd3 binds to the promoter of the Casr gene. Assessment of CaSR signalling showed that JQ1+ significantly reduced Ca2+e-mediated increases in intracellular calcium (Ca2+i) mobilisation and cAMP signalling. However, the CaSR-negative allosteric modulator, NPS-2143, was unable to reduce AtT20 cell proliferation, indicating that reducing CaSR expression rather than activity is likely required to reduce pituitary cell proliferation. Thus, these studies demonstrate that reducing CaSR expression may be a viable option in the treatment of pituitary tumours. Moreover, current strategies to reduce CaSR activity, rather than protein expression for cancer treatments, may be ineffective.


Association of MAPT subhaplotypes with clinical and demographic features in Parkinson's disease.
To determine whether distinct microtubule-associated protein tau MAPT H1 subhaplotypes are associated with clinical and demographic features in Parkinson's disease. A retrospective cohort study included 855 unrelated Caucasian patients with Parkinson's disease who were seen by Movement Disorder specialists at the Mayo Clinic Florida between 1998 and 2016. The primary outcome measures were specific demographic and clinical features of Parkinson's disease, including age at onset, disease progression, survival, motor signs, dementia, dystonia, dyskinesia, autonomic dysfunction, impulse control disorder, psychiatric features, REM sleep behavior disorder, restless legs syndrome, and Parkinson's disease subtype. Specific clinical features were measured at the initial visit and most recent visit. These outcomes were assessed for association with MAPT H1 subhaplotypes, which were defined by six haplotype tagging variants. Median onset age was 64 years (range: 22-94 years); 548 (64%) of patients were male. Significant associations (P < 0.0029) were observed between MAPT H1b and orthostatic hypotension (OR = 1.72, P = 0.001); between H1j and rest tremor (OR = 0.15; P < 0.001) as well as REM sleep behavior disorder (OR = 3.87, P < 0.001); between H1r and bradykinesia (OR = 0.11; P < 0.001); and between H1v and restless legs syndrome (OR = 4.02, P = 0.002). Four MAPT H1 subhaplotypes, but not the H2 haplotype, were significantly associated with specific clinical features in Parkinson's disease. MAPT haplotypic structure may explain some of the phenotypic variability in disease. Replication of our findings will be critical to fully resolve the Parkinson's disease risk association signal at Chr17q21.


Protective role of Phyllantus niruri extract in doxorubicin-induced myocardial toxicity in rats.
To investigate the effect of the aqueous extract of Phyllanthus niruri (Aq.E.PN) against doxorubicin (Dox)-induced myocardial toxicity in rats. Cardiotoxicity was produced by Dox administration (15 mg/kg for 2 weeks). Aq.E PN (200 mg/kg, orally) was administered as pretreatment for 2 weeks alternated with Dox for the next 2 weeks. The general observations, mortality, histopathology, biomarker enzymes like lactate dehydrogenase (LDH), creatinine phosphokinase (CPK) and alkaline phosphatase, diagnostic enzyme markers like aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and antioxidants such as glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) were monitored after 3 weeks of the last dose. Pretreatment with the Aq.E.PN significantly (P < 0.01) protected the myocardium from the toxic effects of Dox by reducing the elevated level of biomarker and diagnostic enzymes like LDH, CPK, AST and ALT to the normal levels. Aq.E PN increased the GSH, SOD and CAT levels and decreased the MDA levels in cardiac tissue. Administration of Dox caused cardiomyopathy associated with an antioxidant deficiency. These results suggest a cardioprotective effect of P. niruri due to its antioxidant properties.


Clinical use of nuclear magnetic resonance imaging in stroke.
There are many positive aspects to the use of MRI in the evaluation of cerebrovascular disease. First, the MR imaging technique appears to be essentially without hazard. It does not rely on ionizing radiation, and no intravenous injections of contrast agent are necessary. MRI exploits the tissue's inherent biophysical characteristics to provide superior contrast. Infarctions are well delineated by MRI, often better and earlier than CT. Because of the lack of MRI signal from bone and thus the lack of transverse artifact from bone often seen with CT, lesions in the posterior fossa are very well visualized. With MRI it is possible to obtain images in the transverse, coronal, and sagittal planes, which provides for good evaluation of lesion size and extent. Arteriovenous malformations have been visualized by MRI, but it is still too early to know whether MRI has any detection capability over CT in this disorder. Subdural hematomas have been well visualized by MRI, including cases of isodense subdurals not visualized by CT. On the other hand, MRI has not proven to have any advantage over CT in the evaluation of intracerebral hemorrhage, hemorrhagic infarction, and subarachnoid hemorrhage. In fact, for detection of intracerebral hemorrhage and subarachnoid hemorrhage, CT may be better at the present time. In chronic infarction the surrounding area of Wallerian degeneration may cause the area of infarction to appear larger than it actually is. Hopefully, with further research into the use of different pulse sequence techniques and with good neuropathological correlation, the present limitations of MRI can be eliminated.(ABSTRACT TRUNCATED AT 250 WORDS)


Polymorphism of the cyclin D1 gene, CCND1, and risk for incident sporadic colorectal adenomas.
Cyclin D1, encoded by the CCND1 gene and activated by the adenomatous polyposis coli-beta-catenin-T-cell factor/lymphoid enhancing factor pathway, induces G(1) to S-phase cell cycle transition, promoting cell proliferation. A recently described codon 242, exon 4, G to A single nucleotide polymorphism (A870G) produces a longer half-life cyclin D1. To investigate whether CCND1 genotype influences risk for colorectal adenoma, we genotyped CCND1 by PCR/RFLP on 161 incident sporadic adenoma cases and 213 controls ages 30-74 years in a North Carolina colonoscopy-based case-control study. At least one polymorphic A allele was found in 68% of cases and 60% of controls. Having an A allele was associated with increased risk for adenoma: the age- and sex-adjusted odds ratio (OR) was 1.5 [95% confidence interval (CI) 1.0-2.4], a finding that was stronger for those whose adenomas were multiple (OR 2.9, 95% CI 1.4-6.0), larger (>or=1 cm; OR 2.4, 95% CI 1.2-4.8), had moderate to severe dysplasia (OR 2.1, 95% CI 1.1-3.8), or were in the right side of the colon (OR 3.6, 95% CI 1.3-10.0). Joint risk factor multivariate analyses revealed stronger positive associations among those who were older (>57 years; OR 2.8, 95% CI 1.4-5.5), male (OR 2.8, 95% CI 1.3-5.7), currently smoked (OR 2.7, 95% CI 1.3-5.7), or currently drank alcohol (OR 2.2, 95% CI 1.2-4.2) if they had an A allele and stronger inverse associations among those who used nonsteroidal anti-inflammatory drugs (OR 0.4, 95% CI 0.2-0.9) or had higher calcium intakes (OR 0.4, 95% CI 0.2-0.9) if they had no A allele. These data support the hypothesis that the CCND1 A870G polymorphism may increase risk for colorectal neoplasms.


MO-C-BRCD-02: Physics of Cancer Cell Migration.
Two-dimensional (2D) in vitro culture systems have for a number of years provided a controlled and versatile environment for the study of cell adhesion and migration, two interrelated cell functions critical to cancer metastasis. However, the organization and functions of focal adhesion proteins in cells embedded in physiologically more relevant 3D matrices is qualitatively and functionally different from their organization and functions on conventional 2D planar substrates. In a 3D, crosslinked, fibrillar collagen matrix, cell migration and protrusion activity are still regulated by focal adhesion proteins, such as pl30Cas, FAK, Zyxin, Vinculin, Talin, and VASP, but differently from the 2D case. This talk will describe the implications of the dependence of focal adhesion rotein-based cellular functions on microenvironmental dimensionality in cancer. We will discuss the implications of this work in cancer metastasis.


Implant osseointegration and the role of microroughness and nanostructures: lessons for spine implants.
The use of spinal implants for spine fusion has been steadily increasing to avoid the risks of complications and donor site morbidity involved when using autologous bone. A variety of fusion cages are clinically available, with different shapes and chemical compositions. However, detailed information about their surface properties and the effects of such properties on osteogenesis is lacking in the literature. Here we evaluate the role of surface properties for spinal implant applications, covering some of the key biological processes that occur around an implant and focusing on the role of surface properties, specifically the surface structure, on osseointegration, drawing examples from other implantology fields when required. Our findings revealed that surface properties such as microroughness and nanostructures can directly affect early cell behavior and long-term osseointegration. Microroughness has been well established in the literature to have a beneficial effect on osseointegration of implants. In the case of the role of nanostructures, the number of reports is increasing and most studies reveal a positive effect from the nanostructures alone and a synergistic effect when combined with microrough surfaces. Long-term clinical results are nevertheless necessary to establish the full implications of surface nanomodifications.


Difference in the interaction of nano-diameter rod and tubular particles with a disclination line in a nematic liquid crystal.
In the presence of a disclination line, inclusions within an aligned nematic liquid crystal (LC) are first attracted and ultimately trapped in it. The kind of orientational distortion created by the inclusions is fundamental in determining the trapping. In the present work, we observe differences in the trapping behaviour, onto a ½ defect line in a nematic LC, of two types of particles both elongated but different in their actual geometry. Even if both types have cylindrical shape, aggregates of Mo6S2I8 nanowires (rod-like shape) and multiwall carbon nanotubes (tubular shape, i.e. hollow) trap differently although still due to deformations induced in the LC director field. Attractive forces are stronger on elongated bundles of nanowires than on similarly sized bundles of multi-wall carbon nanotubes. The reason is the difference in the attraction forces originating from different types of distortions of the LCs. The hollow and the full cylinders are not homotopically equivalent and this inequivalence holds also for the liquid crystal around them. The nanowires induce defects in the LC close-by their surfaces as shown for microrods, topologically equivalent to spheres. In contrast, multi-wall carbon nanotubes, being hollow, do not form defects close to their ends. However, the tubes are strongly bent and the strong planar anchoring of LC at the surface induces deformation in the LC enabling attraction forces with the defect line. HiPco single wall carbon nanotubes could not be trapped because their bundles looked much straighter and smaller than the ones of MWCNTs and thus neither defects nor standard strong deformations are expected. In conclusion, even if the shape of both types of particles is cylindrical, the topological difference between rods and tubes has profound consequences on the physical behaviour and on the presence and type of defect-mediated nematic attraction forces.


Children's screen viewing is related to psychological difficulties irrespective of physical activity.
We hypothesized that greater screen use would be associated with greater psychological difficulties and that children with high levels of screen entertainment use and low levels of physical activity would have the most-negative psychological profiles. Participants were 1013 children (age, mean ± SD: 10.95 ± 0.41 years), who self-reported average daily television hours and computer use and completed the Strengths and Difficulties Questionnaire. Sedentary time (minutes per day with <100 cpm) and moderate/vigorous physical activity (MVPA) (minutes with ≥ 2000 cpm) were measured by using accelerometers. Multivariate regression models examined the association between television viewing, computer use, sedentary time, and Strengths and Difficulties Questionnaire scores, with adjustment for MVPA, age, gender, level of deprivation, and pubertal status. Greater television and computer use were related to higher psychological difficulty scores after adjustment for MVPA, sedentary time, and confounders. However, sedentary time was inversely related to psychological difficulties after adjustment. Children who spent >2 hours per day watching television or using a computer were at increased risk of high levels of psychological difficulties (television, odds ratio [OR]: 1.61 [95% confidence interval [CI]: 1.20-2.15]; computer, OR: 1.59 [95% CI: 1.32-1.91]), and this risk increased if the children also failed to meet physical activity guidelines (television, OR: 1.70 [95% CI: 1.09-2.61]; computer, OR: 1.81 [95% CI: 1.02-3.20]). Both television viewing and computer use are important independent targets for intervention for optimal well-being for children, irrespective of levels of MVPA or overall sedentary time.


Facilitating access to emotions: neural signature of EMDR stimulation.
Eye Movement Desensitisation and Reprocessing (EMDR) is a method in psychotherapy effective in treating symptoms of posttraumatic stress disorder. The client attends to alternating bilateral visual, auditory or sensory stimulation while confronted with emotionally disturbing material. It is thought that the bilateral stimulation as a specific element of EMDR facilitates accessing and processing of negative material while presumably creating new associative links. We hypothesized that the putatively facilitated access should be reflected in increased activation of the amygdala upon bilateral EMDR stimulation even in healthy subjects. We investigated 22 healthy female university students (mean 23.5 years) with fMRI. Subjects were scanned while confronted with blocks of disgusting and neutral picture stimuli. One third of the blocks was presented without any additional stimulation, one third with bilateral simultaneous auditory stimulation, and one third with bilateral alternating auditory stimulation as used in EMDR. Contrasting disgusting vs. neutral picture stimuli confirmed the expected robust effect of amygdala activation for all auditory stimulation conditions. The interaction analysis with the type of auditory stimulation revealed a specific increase in activation of the right amygdala for the bilateral alternating auditory stimulation. Activation of the left dorsolateral prefrontal cortex showed the opposite effect with decreased activation. We demonstrate first time evidence for a putative neurobiological basis of the bilateral alternating stimulation as used in the EMDR method. The increase in limbic processing along with decreased frontal activation is in line with theoretical models of how bilateral alternating stimulation could help with therapeutic reintegration of information, and present findings may pave the way for future research on EMDR in the context of posttraumatic stress disorder.


Serial position memory in the visual-spatial domain: reconstructing sequences of unfamiliar faces.
In two studies we presented pictures of unfamiliar faces one at a time, then presented the complete set at test and asked for serial reconstruction of the order of presentation. Serial position functions were similar to those found with verbal materials, with considerable primacy and one item recency, position errors that were mainly to the adjacent serial position, a visual similarity effect, and effects of articulatory suppression that did not interact with the serial position effect or with the similarity effect. Serial position effects were found when faces had been seen for as little as 300 ms and after a 6-s retention interval filled with articulatory suppression. Serial position effects found with unfamiliar faces are not based on verbal encoding strategies, and important elements of serial memory may be general across modalities.


Lower in vivo mutation rate of human immunodeficiency virus type 1 than that predicted from the fidelity of purified reverse transcriptase.
The level of genetic variation of human immunodeficiency virus type 1 (HIV-1), a member of the lentivirus genus of the Retroviridae family, is high relative to that of retroviruses in some other genera. The high error rates of purified HIV-1 reverse transcriptase in cell-free systems suggest an explanation for this high genetic variation. To test whether the in vivo rate of mutation during reverse transcription of HIV-1 is as high as predicted by cell-free studies, and therefore higher than that rates of mutation of retroviruses in other genera, we developed an in vivo assay for detecting forward mutations in HIV-1, using the lacZ alpha peptide gene as a reporter for mutations. This system allows the rates and types of mutations that occur during a single cycle of replication to be studied. We found that the forward mutation rate for HIV-1 was 3.4 x 10(-5) mutations per bp per cycle. Base substitution mutations predominated; G-to-A transition mutations were the most common base substitution. The in vivo mutation rates for HIV-1 are three and seven times higher than those previously reported for two other retroviruses, spleen necrosis virus and bovine leukemia virus, respectively. In contrast, our calculated in vivo mutation rate for HIV-1 is about 20-fold lower than the error rate of purified HIV-1 reverse transcriptase, with the same target sequence. This finding indicates that HIV-1 reverse transcription in vivo is not as error prone as predicted from the fidelity of purified reverse transcriptase in cell-free studies. Our data suggest that the fidelity of purified HIV-1 reverse transcriptase may not accurately reflect the level of genetic variation in a natural infection.


Selection of chemotaxis mutants of Dictyostelium discoideum.
A method has been developed for the efficient selection of chemotaxis mutants of Dictyostelium discoideum. Mutants defective in the chemotactic response to folate could be enriched up to 30-fold in one round of selection using a chamber in which a compartment that contained the chemoattractant was separated by a sandwich of four nitrocellulose filters from a compartment that contained buffer. Mutagenized cells were placed in the center of the filter layer and exposed to the attractant gradient built up between the compartments for a period of 3-4 h. While wild-type cells moved through the filters in a wave towards the compartment that contained attractant, mutant cells remained in the filter to which they were applied. After several repetitions of the selection procedure, mutants defective in chemotaxis made up 10% of the total cell population retained in that filter. Mutants exhibiting three types of alterations were collected: motility mutants with either reduced speed of movement, or altered rates of turning; a single mutant defective in production of the attractant-degrading enzyme, folate deaminase; and mutants with normal motility but reduced chemotactic responsiveness. One mutant showed drastically reduced sensitivity in folate-induced cGMP production. Morphogenetic alterations of mutants defective in folate chemotaxis are described.