Bowel cancer screening in an English prison: a qualitative service evaluation.
Bowel cancer screening home-testing kits are offered every two years to individuals aged 60-74 years in the United Kingdom (UK), with prisoners eligible for screening in the same way as the general population. There are currently major changes planned to the bowel cancer screening programme in England, with the transition to the single-sample faecal immunochemical test (FIT) and the planned lowering of the age limit from 60 to 50 years. In this project, we aimed to explore processes and beliefs around bowel cancer screening in an English prison. This is a qualitative study. Semistructured qualitative interviews were conducted with eight prisoners and four staff members in a male prison in North West England. Data were analysed via thematic analysis. Promoting and impeding factors to screening were identified. There was high willingness amongst prisoners to be screened for bowel cancer, with screening seen as important and having benefits for the individual and healthcare system. However, there was often low awareness of screening and there were psychological challenges associated with screening. Prison healthcare staff were widely respected and were a motivator to accept screening, with prisoners viewing prison as a good opportunity to access health care. Despite this, prison life was characterised by competing priorities, with security taking precedence, and screening sometimes a low priority for staff and prisoners. There were also considerable logistical challenges to delivering bowel cancer screening in a prison, and the system was not comparable with that in the community. Providing good-quality understandable information, though challenging, was key. This is the first project to explore entering the bowel cancer screening programme in UK prisons, and promoting and impeding factors to the take-up of screening have been identified. This information could be practically used by local commissioners and policymakers to aid the design of staff and prisoner interventions to maximise uptake of bowel cancer screening in prisons. As the planned changes to bowel cancer screening in England are likely to increase the number of eligible prisoners, this work could also be used to inform any service reconfiguration required to accommodate these changes.


Predictable evolution towards larger brains in birds colonizing oceanic islands.
Theory and evidence suggest that some selective pressures are more common on islands than in adjacent mainland habitats, leading evolution to follow predictable trends. The existence of predictable evolutionary trends has nonetheless been difficult to demonstrate, mainly because of the challenge of separating in situ evolution from sorting processes derived from colonization events. Here we use brain size measurements of >1900 avian species to reveal the existence of one such trend: increased brain size in island dwellers. Based on sister-taxa comparisons and phylogenetic ancestral trait estimations, we show that species living on islands have relatively larger brains than their mainland relatives and that these differences mainly reflect in situ evolution rather than varying colonization success. Our findings reinforce the view that in some instances evolution may be predictable, and yield insight into why some animals evolve larger brains despite substantial energetic and developmental costs.


Coagulation and mental disorders.
The neurovascular unit is a key player in brain development, homeostasis, and pathology. Mental stress affects coagulation, while severe mental illnesses, such as recurrent depression and schizophrenia, are associated with an increased thrombotic risk and cardiovascular morbidity. Evidence indicates that the hemostatic system is involved to some extent in the pathogenesis, morbidity, and prognosis of a wide variety of psychiatric disorders. The current review focuses on emerging data linking coagulation and some psychiatric disorders.


The competencies required by professional hospice palliative care spiritual care providers.
The Canadian Hospice Palliative Care Association (2002) identifies spiritual care of the dying and their families as a core service for Hospice Palliative Care programs. Yet, until the Spiritual Care Development Initiative of the Canadian Pallium Project, there was no published literature indicating systematic profiling of occupationally relevant core competencies or competency-based training programs specific to this specialized field of practice. This article describes a Canadian Community of Practice process to develop an occupational analysis-based competency profile for the Professional Hospice Palliative Care Spiritual Care Provider utilizing a modified Developing a Curriculum (DACUM) methodology. Competency profiles are important contributions to the development of curricula to train care providers who are recognized by other professions and by institutions as possessing the requisite theoretical and clinical expertise, particularly in academic tertiary care settings.


PQBP-1/Npw38, a nuclear protein binding to the polyglutamine tract, interacts with U5-15kD/dim1p via the carboxyl-terminal domain.
PQBP-1 was identified as a binding protein to the polyglutamine tract present in various transcription-related factors and causative genes for neurodegenerative disorders. This novel gene contains at least two functional domains, WW domain and carboxyl-terminal domain (CTD), strictly conserved beyond species. Although human PQBP-1 additionally contains the polar amino acid-rich domain by which it binds to the polyglutamine tract, genuine physiological function(s) have not been clarified. In this study, we showed that U5-15kD, human homologue of fission yeast dim1p, is a partner molecule of PQBP-1 binding to CTD. This finding suggests physiological functions of PQBP-1 in splicing, cell cycle, and ubiquitination, through which we can speculate the pathological roles of PQBP-1 in triplet repeat diseases.


Transcriptional and epigenetic regulation of interleukin-2 gene in activated T cells by morphine.
Chronic morphine inhibits interleukin-2 (IL-2) at both the transcriptional and protein synthesis levels. The molecular mechanisms by which morphine decreases IL-2 are not fully understood. The production of IL-2 is tightly regulated by several transcription factors that bind to the IL-2 promoter. Herein, we show that chronic morphine treatment results in an increase in cAMP levels with a concurrent up-regulation of the cAMP inducible repressor inducible cAMP early repressor (ICER)/cAMP response element modulator (CREM) and down-regulation of p-cAMP-response element-binding protein (CREB) in activated T cells. Furthermore, ICER competes for p-CREB binding to the cAMP-responsive elements (CREs) site. This leads to the uncoupling of CBP/p300 thereby abrogating IL-2 transcription. Overexpression of either antisense CREM or CREB plasmid rescued morphine-induced inhibition of IL-2 promoter activity and protein production. In addition, we also found that chronic morphine treatment inhibited the acetylation and trimethylation of histones and decreased both DNA demethylation and accessibility of the IL-2 promoter. These findings suggest that chronic morphine treatment may function through both transcriptional and epigenetic mechanisms to inhibit IL-2 production.


Localizing P300 generators in high-density event- related potential with fMRI.
To assess the effects of stimulus context on the P300 component, an eight-orientation Landolt ring task was introduced. As the stimulus context of this task differs from the traditional two-stimulus oddball paradigm, the purpose here was to apply EEG/fMRI integration to investigate the localization and activities of the P300 generators involved with this task. Ten healthy subjects performed the visual P300 task while fMRI and 64-channel ERP data were acquired. The voltage topographical maps of the P300 component were calculated and analyzed for the main activation foci. Furthermore, constraints from fMRI were used to identify the source activities of visual P300 ERP. Analysis of the hemodynamic response to the visual target stimuli revealed a distributed network of neural sources in the bilateral parietal lobules, middle and inferior frontal gyrus, precentral and postcentral cortex, and anterior cingulate gyrus. The analysis particularly showed preponderant activations of the bilateral superior parietal lobules. In this target detection design, two distinct P300 peaks were observed in the dipole waveforms, the bilateral prefrontal and the right inferior parietal dipole waveforms displayed the higher peak at short latency, while the four parietal, the anterior cingulate, and the temporal dipole waveforms had the higher peak at long latency. Compared with the classical oddball paradigm, the amplitude decreased in this study, which might be related to its particular stimulus context. The source technique was utilized to yield a realistic 11-dipole model and distinguish the anatomical generators of early and late components of the P300 response.


Sterol dynamics during endocytic trafficking in Arabidopsis.
Sterols are lipids found in membranes of eukaryotic cells. Functions of sterols have been demonstrated for various cellular processes including endocytic trafficking in animal, fungal, and plant cells. The ability to visualize sterols at the subcellular level is crucial to understand sterol distribution and function during endocytic trafficking. In plant cells, the polyene antibiotic filipin is the most extensively used tool for the specific detection of fluorescently labeled 3-β-hydroxysterols in situ. Filipin can to some extent be used to track sterol internalization in live cells, but this application is limited, due to the inhibitory effects filipin exerts on sterol-dependent endocytosis. Nevertheless, filipin-sterol labeling can be performed on aldehyde-fixed cells which allows for sterol detection in endocytic compartments. This approach can combine studies correlating sterol distribution with experimental manipulations of endocytic trafficking pathways. Here, we describe step-by-step protocols and troubleshooting for procedures on live and fixed cells to visualize sterols during endocytic trafficking. We also provide a detailed discussion of advantages and limitations of both methods. Moreover, we illustrate the use of the endocytic recycling inhibitor brefeldin A and a genetically modified version of one of its target molecules for studying endocytic sterol trafficking.


Standards for the assessment of visual evoked potentials in an ethnically heterogeneous adult population.
Visual evoked potentials (VEPs) were recorded by chequerboard pattern-reversal stimulation in 276 normal adult subjects aged 15-73 years. The sample comprised comparable white, coloured and black groups of both sexes. Significantly shorter latencies of the major positive component were found in both male and female black subjects compared with those in whites. No significant differences were found between the amplitudes in the different population groups, but females in each population group showed significantly higher amplitudes of the major positive component than males. There was a close relationship between latency and amplitude for both right and left eyes. Except in coloured females, a non-linear relationship of the major positive component with age was demonstrated, the shortest latency being at about 35 years of age. It is suggested that the following criteria be used in the evaluation of VEP recordings: latency as well as the difference between latencies and amplitudes of the two sides of the major positive component (P1) as compared with standardized values for ethnic, sex and age groups.


Sociability and synapse subtype-specific defects in mice lacking SRPX2, a language-associated gene.
The FoxP2 transcription factor and its target genes have been implicated in developmental brain diseases with a prominent language component, such as developmental verbal dyspraxia and specific language impairment. How FoxP2 affects neural circuitry development remains poorly understood. The sushi domain protein SRPX2 is a target of FoxP2, and mutations in SRPX2 are associated with language defects in humans. We have previously shown that SRPX2 is a synaptogenic protein that increases excitatory synapse density. Here we provide the first characterization of mice lacking the SRPX2 gene, and show that these mice exhibit defects in both neural circuitry and communication and social behaviors. Specifically, we show that mice lacking SRPX2 show a specific reduction in excitatory VGlut2 synapses in the cerebral cortex, while VGlut1 and inhibitory synapses were largely unaffected. SRPX2 KO mice also exhibit an abnormal ultrasonic vocalization ontogenetic profile in neonatal pups, and reduced preference for social novelty. These data demonstrate a functional role for SRPX2 during brain development, and further implicate FoxP2 and its targets in regulating the development of vocalization and social circuits.


Immunologic aspects in the pathogenesis and treatment of immune thrombocytopenic purpura in children.
Immune thrombocytopenic purpura (ITP) is either a transient or a chronic bleeding disorder characterized by an unbalanced immune response due to inflammation (ie, infection, autoimmunity). There is new evidence arguing for an immune-mediated mechanism operative in ITP-increased HLA-DR expression, defects in cellular and humoral immunity, and specific autoantibody production. Moreover, different forms of ITP may be associated with specific alterations in the immune response. New immunosupportive modalities emphasize biologic interventions as a therapeutic strategy for autoimmune disorders. Practical guidelines for the diagnosis and treatment of ITP were recently established by the American Society of Hematology. These recommendations will substantially influence patient care and future research.


Composition and expression of conserved microRNA genes in diploid cotton (Gossypium) species.
MicroRNAs are ubiquitous in plant genomes but vary greatly in their abundance within and conservation among plant lineages. To gain insight into the evolutionary birth/death dynamics of microRNA families, we sequenced small RNA and 5'-end PARE libraries generated from two closely related species of Gossypium. Here, we demonstrate that 33 microRNA families, with similar copy numbers and average evolutionary rates, are conserved in the two congeneric cottons. Analysis of the presence/absence of these microRNA families in other land plants sheds light on their depth of phylogenetic origin and lineage-specific loss/gain. Conserved microRNA families in Gossypium exhibit a striking interspecific asymmetry in expression, potentially connected to relative proximity to neighboring transposable elements. A complex correlated expression pattern of microRNA target genes with their controlling microRNAs indicates that possible functional divergence of conserved microRNA families can also exist even within a single plant genus.


Acupuncture in the neonatal intensive care unit-using ancient medicine to help today's babies: a review.
Acupuncture has been used for thousands of years in Eastern medicine for a variety of conditions and illnesses, including pain. Neonatal intensive care, on the other hand, is a relatively new branch of medicine that has emerged as the pivotal influence in increasing survival of critically ill newborn infants only within the last 50 years. Unfortunately, pain is an inevitable part of treatment in a neonatal intensive care unit (NICU). The control and prevention of pain remains a major issue for clinicians despite recognition and understanding of the myriad of short- and long-term problems that are associated with both pain and its treatment within the NICU environment. In this review, we examine the potential role of acupuncture to decrease and treat pain in babies requiring neonatal intensive care and discuss future therapeutic and research implications for the use of this ancient therapy within the modern environment of the NICU.


Social cues to joint actions: the role of shared goals.
In daily life, we do not just move independently from how others move. Rather, the way we move conveys information about our cognitive and affective attitudes toward our conspecifics. However, the implicit social substrate of our movements is not easy to capture and isolate given the complexity of human interactive behaviors. In this perspective article we discuss the crucial conditions for exploring the impact of "interpersonal" cognitive/emotional dimensions on the motor behavior of individuals interacting in realistic contexts. We argue that testing interactions requires one to build up naturalistic and yet controlled scenarios where participants reciprocally adapt their movements in order to achieve an overarching "shared goal." We suggest that a shared goal is what singles out real interactions from situations where two or more individuals contingently but independently act next to each other, and that "interpersonal" socio-emotional dimensions might fail to affect co-agents' behaviors if real interactions are not at place. We report the results of a novel joint-grasping task suitable for exploring how individual sub-goals (i.e., correctly grasping an object) relate to, and depend from, the representation of "shared goals."


The replicative helicase MCM recruits cohesin acetyltransferase ESCO2 to mediate centromeric sister chromatid cohesion.
Chromosome segregation depends on sister chromatid cohesion which is established by cohesin during DNA replication. Cohesive cohesin complexes become acetylated to prevent their precocious release by WAPL before cells have reached mitosis. To obtain insight into how DNA replication, cohesion establishment and cohesin acetylation are coordinated, we analysed the interaction partners of 55 human proteins implicated in these processes by mass spectrometry. This proteomic screen revealed that on chromatin the cohesin acetyltransferase ESCO2 associates with the MCM2-7 subcomplex of the replicative Cdc45-MCM-GINS helicase. The analysis of ESCO2 mutants defective in MCM binding indicates that these interactions are required for proper recruitment of ESCO2 to chromatin, cohesin acetylation during DNA replication, and centromeric cohesion. We propose that MCM binding enables ESCO2 to travel with replisomes to acetylate cohesive cohesin complexes in the vicinity of replication forks so that these complexes can be protected from precocious release by WAPL Our results also indicate that ESCO1 and ESCO2 have distinct functions in maintaining cohesion between chromosome arms and centromeres, respectively.


Aggregation in malaria parasites places limits on mosquito infection rates.
Gametocytes are responsible for the transmission of malaria parasites, Plasmodium spp., from man to mosquito. Although transmission success, as measured by the proportion of mosquitoes infected, generally increases with gametocyte density, the proportion of parasites that are gametocytes is always paradoxically only a few percent of the asexual blood parasites. To address this paradox, we analyse transmission data sets from an urban and an adjacent rural setting in Cameroon to elucidate whether there are discernable lower and upper limits to Plasmodium falciparum gametocyte density that are linked to transmission success. We find that there exists a lower gametocyte density at which mosquito infection rates considerably increase. In addition, we identify upper gametocyte densities at which mosquito infection rates level off. Greatest increases in infection rates occur at low gametocyte densities and coincide with maximum oocyst aggregation within the infected mosquito population. This aggregated oocyst distribution remains despite increases in gametocyte density and ever-decreasing gains in mosquito infection rates. There is increasing suggestion that malaria parasites have evolved sex allocation strategies to ensure transmission in response to a changing, transmission-blocking environment. Here transmission-blocking immunity is proposed not only to ensure low density gametocyte transmission success but also to impose upper limits on transmission success.


The primary cilium of connective tissue cells: imaging by multiphoton microscopy.
Although the role of the primary cilium as a sensory organelle in epithelial cells has been elucidated significantly over the past decade, the function of primary cilia in connective tissue cells has been studied less extensively. Primary cilia have been implicated as mechanotransducers in connective tissues, but the mechanisms by which the cells sense loads and convert them to biochemical signals for tissue formation and adaptation are poorly understood. Before hypotheses regarding the function of the primary cilium in connective tissue cells can be tested, methods for quantitation of incidence as well as three-dimensional visualization of primary cilia with respect to the extracellular matrix (ECM) are needed. The objective of this study was to develop a rapid method for visualizing primary cilia in their native ECM in a wide range of connective tissues. Whole-mount immunohistochemical and multiphoton microscopy techniques were developed to simultaneously image primary cilia, cell nuclei, and collagen and their relationships to each other in situ. Axonemes of primary cilia projecting into the ECM were successfully visualized in thick sections of growth plate cartilage, tendon, ligament, meniscus, intervertebral disc, and perichondrium. These methodologies will allow analysis of the incidence and three-dimensional orientation of primary cilia and enable investigation of the role of primary cilia in normal and pathological growth and adaptation in a variety of musculoskeletal tissues.


Purification and characterization of the DNA polymerase alpha associated exonuclease: the RTH1 gene product.
We report here the purification and mechanistic characterization of a 5'-3' exonuclease associated with DNA polymerase alpha from the yeast Saccharomyces cerevisiae. Earlier, we identified a 5' --> 3' exonuclease activity that copurified with yeast DNA polymerase alpha-primase in a multiprotein complex [Biswas, E. E., et al. (1993) Biochemistry, 32, 3020-3027]. Peptide sequence analysis of the purified 47 kDa exonuclease was carried out, and the peptide sequence was found to be identical to the S. cerevisiae gene YKL510 encoded polypeptide, which is also known as yeast RAD2 homolog 1 or RTH1 nuclease. The native exonuclease also had strong flap endonuclease activity similar to that observed with RTH1 nuclease and homologous yeast (RAD2) and mammalian enzymes. During our studies, we have discovered certain unique features of the mechanism of action of the native RTH1 nuclease. Studies presented here indicated that the exonuclease had specific pause sites during its 5'-3' exonuclease nucleotide excision. These pause sites were easily detected with long (approximately 50 bp) oligonucleotide substrates during exonucleolytic excision by the formation of a discontinuous ladder of excision product. We have further analyzed the mechanism of generation of the pause sites, as they could occur through a number of different pathways. Alignment of the pause sites with the nucleotide sequence of the oligonucleotide substrate indicated that the pause sites were dependent on the nucleotide sequence. Our analysis revealed that RTH1 nuclease pauses predominantly at G:C rich sequences. With poly(dA):oligo(dT)50 as substrate, the exonucleolytic products formed a continuous ladder with no evidence of pausing. The G:C rich DNA sequences are thermodynamically more stable than the A:T rich sequences, which may be in part responsible for pausing of the RTH1 5' --> 3' exonuclease at these sites.


Engineering human hematopoietic environments through ossicle and bioreactor technologies exploitation.
The bone marrow microenvironment contains cellular niches that maintain the pool of hematopoietic stem and progenitor cells and support hematopoietic maturation. Malignant hematopoietic cells also co-opt normal cellular interactions to promote their own growth and evade therapy. In vivo systems used to study human hematopoiesis have been developed through transplantation into immunodeficient mouse models. However, incomplete cross-compatibility between the murine stroma and transplanted human hematopoietic cells limits the rate of engraftment and the study of relevant interactions. To supplement in vivo xenotransplantation models, complementary strategies have recently been developed, including the use of three-dimensional human bone marrow organoids in vivo, generated from bone marrow stromal cells seeded onto osteo-inductive scaffolds, as well as the use of ex vivo bioreactor models. These topics were the focus of the Spring 2020 International Society for Experimental Hematology New Investigator webinar. We review here the latest advances in generating humanized hematopoietic organoids and how they allow for the study of novel microenvironmental interactions.


The Health-Related Quality of Life for Patients with Myalgic Encephalomyelitis / Chronic Fatigue Syndrome (ME/CFS).
Myalgic encephalomyelitis (ME)/chronic fatigue syndrome (CFS) is a common, severe condition affecting 0.2 to 0.4 per cent of the population. Even so, no recent international EQ-5D based health-related quality of life (HRQoL) estimates exist for ME/CFS patients. The main purpose of this study was to estimate HRQoL scores using the EQ-5D-3L with Danish time trade-off tariffs. Secondary, the aims were to explore whether the results are not influenced by other conditions using regression, to compare the estimates to 20 other conditions and finally to present ME/CFS patient characteristics for use in clinical practice. All members of the Danish ME/CFS Patient Association in 2013 (n=319) were asked to fill out a questionnaire including the EQ-5D-3L. From these, 105 ME/CFS patients were identified and gave valid responses. Unadjusted EQ-5D-3L means were calculated and compared to the population mean as well as to the mean of 20 other conditions. Furthermore, adjusted estimates were calculated using ordinary least squares (OLS) regression, adjusting for gender, age, education, and co-morbidity of 18 self-reported conditions. Data from the North Denmark Health Profile 2010 was used as population reference in the regression analysis (n=23,392). The unadjusted EQ-5D-3L mean of ME/CFS was 0.47 [0.41-0.53] compared to a population mean of 0.85 [0.84-0.86]. The OLS regression estimated a disutility of -0.29 [-0.21;-0.34] for ME/CFS patients in this study. The characteristics of ME/CFS patients are different from the population with respect to gender, relationship, employment etc. The EQ-5D-3L-based HRQoL of ME/CFS is significantly lower than the population mean and the lowest of all the compared conditions. The adjusted analysis confirms that poor HRQoL of ME/CFS is distinctly different from and not a proxy of the other included conditions. However, further studies are needed to exclude the possible selection bias of the current study.


Forskolin enhancement of opioid currents in rat locus coeruleus neurons.
1. Opioids are known to hyperpolarize all neurons in the nucleus locus coeruleus (LC) and to inhibit adenylyl cyclase. Recent work has shown that activation of adenylyl cyclase with forskolin increased the amplitude of the opioid hyperpolarization in LC cells. The aim of the present study was to determine the mechanism of this augmented hyperpolarization. 2. Agonist-induced currents were studied in LC cells in brain slices using both intracellular and whole cell recordings. Forskolin increased the amplitude of mu-opioid- and alpha 2-adrenoceptor-mediated currents by approximately 30% of control measured at -60 mV. This effect of forskolin was dependent on the concentration having a threshold of approximately 1 microM and a peak effect at approximately 30 microM. Dideoxyforskolin (30 microM) caused a small reduction (-52 +/- 28 pA, mean +/- SE) in the amplitude of the opioid current. 3. Forskolin increased the agonist current in the outward direction over the entire potential range between -140 and -50 mV when recordings were made from neurons in cells recorded from slices cut in the horizontal plane. This augmented current produced a shift of the apparent reversal potential to more negative values. 4. Both the forskolin augmentation of the opioid current and the opoid current itself were reduced when the space clamp was improved by cutting the slice in the coronal plane, increasing the extracellular potassium concentration, and treating the slice with carbenoxolone. In addition, forskolin did not change the reversal potential of the opoid current. When expressed as a percentage change from control, forskolin had no significant effect on the opioid current in carbenoxolone (-13 +/- 13%) but produced a small augmentation in high extracellular potassium (15 +/- 4%) and coronoal slices (31 +/- 12%). 5. Two models were tested to explain the action of forskolin, one where cells are coupled electronically by a forskolin-sensitive conductance (coupled-cell model) and a second where opioids mediate an inhibition of a forskolin-induced cation conductance (2-conductance model). The experimental results were fit well only by the coupled-cell model, which predicted that the opioid/forskolin interaction is indirect and occurs primarily in response to forskolin increasing the degree of electrotonic coupling between LC neurons. The consequence of increased coupling would be to augment synchronous activity within the nucleus.


Patient-Reported and Objectively Measured Function Before and After Reverse Shoulder Arthroplasty.
Documenting functional outcomes after reverse shoulder arthroplasty (RSA) is critical to advancing patient care. The interplay been self-reported and objectively measured outcome measures has not been widely described. The utilization of wearable devices to document upper extremity limb activity is a new approach for objectively measuring outcomes. Therefore, the purpose of this study was to evaluate changes in pain, and self-reported function and objectively measured limb activity after RSA. We also assessed the influence of pain on self-reported function and objectively measured limb activity to determine the impact of pain on outcomes after RSA. This study implemented a prospective, repeated-measures design. Fourteen patients undergoing RSA underwent testing before surgery, and 2 and 12 months after surgery. Patient-reported instruments included pain, Disabilities of Arm, Shoulder and hands (DASH), and physical component summary (PCS) of the 36-Item Short Form Health Survey. Objective limb activity (mean activity value, m/s/min epoch; inactive time, %; low activity, %; and high activity, %) was captured with triaxial accelerometers worn on the upper and lower arm. A repeated-measures ANOVA tested for differences across time. The Spearman rank-order correlation was calculated to evaluate the influence of pain on DASH, PCS scores, and mean limb activity. Patient-reported measures improved after surgery (pain, P < .01; DASH, P < .01; PCS, P = .01). No change in limb activity was found at 1 year compared with preoperative values for mean (forearm, P = 1.00; arm, P = .36), inactivity (forearm, P = .33; arm, P = .22), low (forearm, P = .77; arm, P = .11) or high (forearm, P = 1.00; arm, P = .20) activity. There was a relationship between pain and DASH scores 1 year after surgery (P = .04) but not before surgery (P = .16), or 2 months after surgery (P = .30). There was no relationship between pain and PCS scores at any time point (preoperative, P = .97; 2 months, P = .21; 1 year, P = .08) nor pain and limb activity (forearm: preoperative, P = .36; 2 months, P = .67; 1 year, P = .16; arm: preoperative, P = .97; 2 months, P = .59; 1 year, P = .51). RSA reduced pain and enhanced patient-perceived function. Objectively measured upper extremity limb activity is not different 1 year after surgery compared with preoperative levels. III.


Gentlest ascent dynamics for calculating first excited state and exploring energy landscape of Kohn-Sham density functionals.
We develop the gentlest ascent dynamics for Kohn-Sham density functional theory to search for the index-1 saddle points on the energy landscape of the Kohn-Sham density functionals. These stationary solutions correspond to excited states in the ground state functionals. As shown by various examples, the first excited states of many chemical systems are given by these index-1 saddle points. Our novel approach provides an alternative, more robust way to obtain these excited states, compared with the widely used ΔSCF approach. The method can be easily generalized to target higher index saddle points. Our results also reveal the physical interest and relevance of studying the Kohn-Sham energy landscape.


STRATEGIES FOR EQUITABLE PHARMACOGENOMIC-GUIDED WARFARIN DOSING AMONG EUROPEAN AND AFRICAN AMERICAN INDIVIDUALS IN A CLINICAL POPULATION.
The blood thinner warfarin has a narrow therapeutic range and high inter- and intra-patient variability in therapeutic doses. Several studies have shown that pharmacogenomic variants help predict stable warfarin dosing. However, retrospective and randomized controlled trials that employ dosing algorithms incorporating pharmacogenomic variants under perform in African Americans. This study sought to determine if: 1) including additional variants associated with warfarin dose in African Americans, 2) predicting within single ancestry groups rather than a combined population, or 3) using percentage African ancestry rather than observed race, would improve warfarin dosing algorithms in African Americans. Using BioVU, the Vanderbilt University Medical Center biobank linked to electronic medical records, we compared 25 modeling strategies to existing algorithms using a cohort of 2,181 warfarin users (1,928 whites, 253 blacks). We found that approaches incorporating additional variants increased model accuracy, but not in clinically significant ways. Race stratification increased model fidelity for African Americans, but the improvement was small and not likely to be clinically significant. Use of percent African ancestry improved model fit in the context of race misclassification.


Interferon signature in patients with STAT1 gain-of-function mutation is epigenetically determined.
STAT1 gain-of-function (GOF) variants lead to defective Th17 cell development and chronic mucocutaneous candidiasis (CMC), but frequently also to autoimmunity. Stimulation of cells with STAT1 inducing cytokines like interferons (IFN) result in hyperphosphorylation and delayed dephosphorylation of GOF STAT1. However, the mechanism how the delayed dephosphorylation exactly causes the increased expression of STAT1-dependent genes, and how the intracellular signal transduction from cytokine receptors is affected, remains unknown. In this study we show that the circulating levels of IFN-α were not persistently elevated in STAT1 GOF patients. Nevertheless, the expression of interferon signature genes was evident even in the patient with low or undetectable serum IFN-α levels. Chromatin immunoprecipitation (ChIP) experiments revealed that the active chromatin mark trimethylation of lysine 4 of histone 3 (H3K4me3), was significantly enriched in areas associated with interferon-stimulated genes in STAT1 GOF cells in comparison to cells from healthy donors. This suggests that the chromatin binding of GOF STAT1 variant promotes epigenetic changes compatible with higher gene expression and elevated reactivity to type I interferons, and possibly predisposes for interferon-related autoimmunity. The results also suggest that epigenetic rewiring may be responsible for treatment failure of Janus kinase 1/2 (JAK1/2) inhibitors in certain patients.


Measuring the Benefits of Mass Vaccination Programs in the United States.
Since the late 1940s, mass vaccination programs in the USA have contributed to the significantly reduced morbidity and mortality of infectious diseases. To assist the evaluation of the benefits of mass vaccination programs, the number of individuals who would have suffered death or permanent disability in the USA in 2014, had mass vaccination never been implemented, was estimated for measles, mumps, rubella, tetanus, diphtheria, pertussis, polio, Haemophilus influenzae type b (Hib), hepatitis B, varicella, and human papillomavirus (HPV). The estimates accounted for mortality and morbidity trends observed for these infections prior to mass vaccination and the impact of advances in standard of living and health care. The estimates also considered populations with and without known factors leading to an elevated risk of permanent injury from infection. Mass vaccination prevented an estimated 20 million infections and 12,000 deaths and permanent disabilities in 2014, including 10,800 deaths and permanent disabilities in persons at elevated risk. Though 9000 of the estimated prevented deaths were from liver cirrhosis and cancer, mass vaccination programs have not, at this point, shown empirical impacts on the prevalence of those conditions. Future studies can refine these estimates, assess the impact of adjusting estimation assumptions, and consider additional risk factors that lead to heightened risk of permanent harm from infection.


p53 and K-ras in radon-associated lung adenocarcinoma.
Mutations in the p53 tumor suppressor gene and the K-ras proto-oncogene are common genetic defects in lung cancer. Analysis of the patterns of damage in these genes may provide important insights into the mechanisms by which environmental mutagens initiate cancer. Previously, our laboratory found that a rare p53 codon 249 mutation (AGG(ARG) to ATG(MET) transversion) was present in 31% of a series of 52 large and squamous cell lung cancers from uranium miners, suggesting that this mutation might be a marker for radon exposure. In the current study, we analyzed 23 lung adenocarcinomas from the same cohort of highly exposed uranium miners. These tumors failed to show the codon 249 transversion, but 9 (39%) of 23 contained 1 or more mutations within hotspots in the K-ras gene. The results suggest that there is a histological tissue-type specificity for the codon 249 mutation; although this mutation was common in squamous and large cell tumors from very highly exposed uranium miners, it is rare in adenocarcinomas from the same cohort of miners.


Association between individual quadriceps muscle volume/enthesis and patello femoral joint cartilage morphology.
The aim of this study was to determine the association between individual quadriceps muscle volumes and the quadriceps enthesis structures and cartilage morphology at the patellofemoral joint (PFJ). We studied 12 cadavers (age 75 ± 5 years). For both legs, individual quadriceps muscles (vastus lateralis (VL), rectus femoris (RF), vastus intermedialis (VI) and vastus medialis (VM)) were dissected and their volumes measured. Cartilage areas at the PFJ were classified using the International Cartilage Repair Society (ICRS) score. Histological sections were evaluated at the quadriceps tendon enthesis (laterally, centrally and medially). Several variables were calculated on the binary images based on two-dimensional analysis. These were apparent bone area (BA) and apparent trabecular thickness (TH). A Spearman rank test was used to determine the strength of correlation between individual quadriceps muscles volume, the structure of the quadriceps tendon enthesis and the ICRS score. The thickness of calcified fibrocartilage tissue was significantly greater in the central part of the enthesis than both medially (P = 0.03) and laterally (P = 0.04). Uncalcified fibrocartilage was significantly thicker laterally (P = 0.04) and centrally (P = 0.02) than medially. Muscle volume was highest (P <0.05) for the VL, followed by the VI, VM and RF. There was no association between total and individual muscle volumes and ICRS or BA. However, there was a strong positive correlation (r = 0.81) between the VL/VM volume ratio and BA ratio (bone volume at the lateral part divided by bone volume at the medial part). There was a moderate positive correlation between VL/VM and ICRS (r = 0.65) and between ICRS and BA ratio (lateral/medial; r = 0.74). Individual and total quadriceps volumes were not correlated with cartilage loss at the PFJ or fibrocartilage thickness. However, both VL/VM and BA ratio (lateral/medial) were positively correlated with ICRS scoring and therefore could be a tool for predicting degree of PFJ osteoarthritis severity.


Proportions of Kaposi sarcoma, selected non-Hodgkin lymphomas, and cervical cancer in the United States occurring in persons with AIDS, 1980-2007.
Given the higher risk of AIDS-defining malignancies that include Kaposi sarcoma (KS), certain non-Hodgkin lymphomas (NHLs), and cervical cancer in persons with human immunodeficiency virus (HIV) infection, the HIV epidemic has likely contributed to the overall numbers of these cancers in the United States. To quantify the proportions of KS, AIDS-defining NHLs, and cervical cancer in the United States that occurred among persons with AIDS from 1980 to 2007. The HIV/AIDS Cancer Match Study (1980-2007) linked data from 16 US HIV/AIDS and cancer registries to identify cases with and without AIDS for KS, AIDS-defining NHLs (ie, diffuse large B-cell lymphoma [DLBCL], Burkitt lymphoma [BL], and central nervous system [CNS] lymphoma), and cervical cancer. Using linked data, we derived cancer rates for persons with and without AIDS. To estimate national counts, the rates were applied to national AIDS surveillance and US Census data. Proportion of AIDS-defining malignancies in the United States occurring in persons with AIDS. In the United States, an estimated 81.6% (95% confidence interval [CI], 81.2%-81.9%) of 83,252 KS cases, 6.0% (95% CI, 5.8%-6.1%) of 351,618 DLBCL cases, 19.9% (95% CI, 18.1%-21.7%) of 17,307 BL cases, 27.1% (95% CI, 26.1%-28.1%) of 27,265 CNS lymphoma cases, and 0.42% (95% CI, 0.37%-0.47%) of 375,452 cervical cancer cases occurred among persons with AIDS during 1980-2007. The proportion of KS and AIDS-defining NHLs in persons with AIDS peaked in the early 1990s (1990-1995: KS, 90.5% [95% CI, 90.2%-90.8%]; DLBCL, 10.2% [95% CI, 9.9%-10.5%]; BL, 27.8% [95% CI, 25.0%-30.5%]; and CNS lymphoma, 48.3% [95% CI, 46.7%-49.8%]; all P < .001 [compared with 1980-1989]) and then declined (2001-2007: KS, 70.5% [95% CI, 68.1%-73.0%]; DLBCL, 4.7% [95% CI, 4.3%-5.2%]; BL, 21.5% [95% CI, 17.7%-25.4%]; and CNS lymphoma, 12.9% [95% CI, 10.5%-15.3%]; all P < .001 [compared with 1990-1995]). The proportion of cervical cancers in persons with AIDS increased over time (1980-1989: 0.11% [95% CI, 0.09%-0.13%]; 2001-2007: 0.71% [95% CI, 0.51%-0.91%]; P < .001). In the United States, the estimated proportions of AIDS-defining malignancies that occurred among persons with AIDS were substantial, particularly for KS and some NHLs. Except for cervical cancer, the proportions of AIDS-defining malignancies occurring among persons with AIDS peaked in the mid-1990s and then declined.


Spectral redemption in clustering sparse networks.
Spectral algorithms are classic approaches to clustering and community detection in networks. However, for sparse networks the standard versions of these algorithms are suboptimal, in some cases completely failing to detect communities even when other algorithms such as belief propagation can do so. Here, we present a class of spectral algorithms based on a nonbacktracking walk on the directed edges of the graph. The spectrum of this operator is much better-behaved than that of the adjacency matrix or other commonly used matrices, maintaining a strong separation between the bulk eigenvalues and the eigenvalues relevant to community structure even in the sparse case. We show that our algorithm is optimal for graphs generated by the stochastic block model, detecting communities all of the way down to the theoretical limit. We also show the spectrum of the nonbacktracking operator for some real-world networks, illustrating its advantages over traditional spectral clustering.


Automotive sulfate emission data.
This paper discusses automotive sulfate emission results obtained by the Office of Mobile Source Air Pollution Control of EPA, General Motors, Ford, Chrysler, and Esso. This work has been directed towards obtaining sulfate emission factors for cars with and without catalyst. While the EPA and Chrysler investigations have found significant sulfate formation in noncatalyst cars, GM, Ford, and Esso have found only trace levels from noncatalyst cars. All of these investigators agree that much higher quantities of sulfate are emitted from catalyst cars. The work done to date shows pelleted catalysts to have much lower sulfate emissions over the low speed-EPA Federal Test Procedures than monolith catalysts. This is probably due to temporary storage of sulfates on the catalyst due to chemical interaction with the alumina pellets. The sulfate compounds are, to a large degree, emitted later under higher speed conditions which result in higher catalyst temperatures which decompose the alumina salt. Future work will be directed towards further elucidation of this storage mechanism as well as determining in detail how factors such as air injection rate and catalyst location affect sulfate emissions.


Athletes' use of exercise imagery during weight training.
Imagery is a cognitive process during which people use their minds to create (or recreate) experiences that are similar to real-life situations. This study examined how college athletes used imagery during weight training. Subjects were 295 Division I (n = 163) and Division II (n = 132) college student athletes (men: n = 138, women: n = 157) who participated in a weight training program as a requirement of their sport. They completed a slightly modified version of the "Weight Lifting Imagery Questionnaire." Results showed that appearance imagery (i.e., images related to the attainment of a fit-looking body) was used and considered the most effective followed by technique imagery (i.e., images related to performing the skill and techniques correctly with good form) and energy imagery (i.e., images related to getting "psyched up" or feeling energized). Other variables that effected imagery use were gender, age, time of season, and levels of motivation. In addition, gender, previous imagery training, and level of motivation had an effect on the perceptions of imagery effectiveness. Confidence in the ability to image was associated with both imagery use and effectiveness, and imagery use and effectiveness were associated with confidence in the weight room. The findings support previous research in exercise imagery that appearance imagery is most used followed by technique and energy imagery and extend them in such a way that strength coaches have practical advice on how to use imagery in a positive way with their athletes. Suggestions about how strength coaches can use imagery with their clients are provided.


Augmentin duo™ in the treatment of childhood blepharokeratoconjunctivitis.
To report the use of Augmentin Duo 400/57 (GlaxoSmithKline, Middlesex, UK) in the treatment of childhood blepharokeratoconjunctivitis (BKC). This is a retrospective interventional case series. The case notes of 7 consecutive patients treated with Augmentin Duo 400/57 for BKC during 18 months were reviewed. Diagnostic criteria for BKC were blepharitis including recurrent chalazia and meibomian gland dysfunction, eyelid margin telangiectasia and facial rosacea, recurrent episodes of chronic red eye, photophobia, watering, punctate superficial keratopathy, corneal neovascularization, and corneal ulcers. Seven children (age range: 6 to 14 years) were diagnosed as having BKC. All children received systemic Augmentin Duo 400/57 and showed considerable improvement within the first month of therapy. Six children had no recurrences during a mean follow-up of 6 months. No patients experienced any side effects from this treatment. Augmentin Duo 400/57 has not previously been reported in the treatment of BKC in children. In this case series, Augmentin Duo 400/57 proved to be at least as effective as current treatments with systemic erythromycin or doxycycline with the advantage of a twice-daily dosage and a superior side-effect profile.


Spatial familial networks to infer demographic structure of wild populations.
In social species, reproductive success and rates of dispersal vary among individuals resulting in spatially structured populations. Network analyses of familial relationships may provide insights on how these parameters influence population-level demographic patterns. These methods, however, have rarely been applied to genetically derived pedigree data from wild populations.Here, we use parent-offspring relationships to construct familial networks from polygamous boreal woodland caribou (Rangifer tarandus caribou) in Saskatchewan, Canada, to inform recovery efforts. We collected samples from 933 individuals at 15 variable microsatellite loci along with caribou-specific primers for sex identification. Using network measures, we assess the contribution of individual caribou to the population with several centrality measures and then determine which measures are best suited to inform on the population demographic structure. We investigate the centrality of individuals from eighteen different local areas, along with the entire population.We found substantial differences in centrality of individuals in different local areas, that in turn contributed differently to the full network, highlighting the importance of analyzing networks at different scales. The full network revealed that boreal caribou in Saskatchewan form a complex, interconnected familial network, as the removal of edges with high betweenness did not result in distinct subgroups. Alpha, betweenness, and eccentricity centrality were the most informative measures to characterize the population demographic structure and for spatially identifying areas of highest fitness levels and family cohesion across the range. We found varied levels of dispersal, fitness, and cohesion in family groups. Synthesis and applications: Our results demonstrate the value of different network measures in assessing genetically derived familial networks. The spatial application of the familial networks identified individuals presenting different fitness levels, short- and long-distance dispersing ability across the range in support of population monitoring and recovery efforts.


Degradation of p57Kip2 mediated by SCFSkp2-dependent ubiquitylation.
The abundance of the cyclin-dependent kinase (CDK) inhibitor p57Kip2, an important regulator of cell cycle progression, is thought to be controlled by the ubiquitin-proteasome pathway. The Skp1/Cul1/F-box (SCF)-type E3 ubiquitin ligase complex SCFSkp2 has now been shown to be responsible for regulating the cellular level of p57Kip2 by targeting it for ubiquitylation and proteolysis. The elimination of p57Kip2 was impaired in Skp2-/- cells, resulting in abnormal accumulation of the protein. Coimmunoprecipitation analysis also revealed that Skp2 interacts with p57Kip2 in vivo. Overexpression of WT Skp2 promoted degradation of p57Kip2, whereas expression of a dominant negative mutant of Skp2 prolonged the half-life of p57Kip2. Mutation of the threonine residue (Thr-310) of human p57Kip2 that is conserved between the COOH-terminal QT domains of p57Kip2 and p27Kip1 prevented the effect of Skp2 on the stability of p57Kip2, suggesting that phosphorylation at this site is required for SCFSkp2-mediated ubiquitylation. Finally, the purified recombinant SCFSkp2 complex mediated p57Kip2 ubiquitylation in vitro in a manner dependent on the presence of the cyclin E-CDK2 complex. These observations thus demonstrate that the SCFSkp2 complex plays an important role in cell-cycle progression by determining the abundance of p57Kip2 and that of the related CDK inhibitor p27Kip1.


Andreas Vesalius (1514-1564) - an unfinished life.
The fame of Andreas Vesalius (1514-1564) rests on his anatomy text, De humani corporis fabrica, regarded as a seminal book in modern medicine. It was compiled while he taught anatomy at Padua, 1537-1543. Some of his findings challenged Galen's writings of the 2c AD, and caused De fabrica to be rejected immediately by classically trained anatomists. At age 29, Vesalius abandoned his studies and over the next two decades served as physician to Emperor Charles V of the Holy Roman Empire (HRE) and later to King Philip II of Spain in Madrid. In 1564, he sought to resume teaching anatomy in Padua, but release from royal service obliged him first to make a pilgrimage to Palestine. During the return voyage to Venice, he became ill and was put ashore alone on an Ionian island Zakynthos, where he died days later at age 50.


Effectiveness of voriconazole in the treatment of Aspergillus fumigatus-associated asthma (EVITA3 study).
IgE sensitization to Aspergillus fumigatus and a positive sputum fungal culture result are common in patients with refractory asthma. It is not clear whether these patients would benefit from antifungal treatment. We sought to determine whether a 3-month course of voriconazole improved asthma-related outcomes in patients with asthma who are IgE sensitized to A fumigatus. Asthmatic patients who were IgE sensitized to A fumigatus with a history of at least 2 severe exacerbations in the previous 12 months were treated for 3 months with 200 mg of voriconazole twice daily, followed by observation for 9 months, in a double-blind, placebo-controlled, randomized design. Primary outcomes were improvement in quality of life at the end of the treatment period and a reduction in the number of severe exacerbations over the 12 months of the study. Sixty-five patients were randomized. Fifty-nine patients started treatment (32 receiving voriconazole and 27 receiving placebo) and were included in an intention-to-treat analysis. Fifty-six patients took the full 3 months of medication. Between the voriconazole and placebo groups, there were no significant differences in the number of severe exacerbations (1.16 vs 1.41 per patient per year, respectively; mean difference, 0.25; 95% CI, 0.19-0.31), quality of life (change in Asthma Quality of Life Questionnaire score, 0.68 vs 0.88; mean difference between groups, 0.2; 95% CI, -0.05 to -0.11), or any of our secondary outcome measures. We were unable to show a beneficial effect of 3 months of treatment with voriconazole in patients with moderate-to-severe asthma who were IgE sensitized to A fumigatus on either the rate of severe exacerbations, quality of life, or other markers of asthma control.


Synthesis of F16 conjugated with 5-fluorouracil and biophysical investigation of its interaction with bovine serum albumin by a spectroscopic and molecular modeling approach.
5-Fluorouracil (5-FU) has been widely used as a chemotherapy agent in the treatment of many types of solid tumors. Investigation of its antimetabolites led to the development of an entire class of fluorinated pyrimidines. However, the toxicity profile associated with 5-FU is significant and includes diarrhea, mucositis, hand-foot syndrome and myelosuppression. In aiming at reducing of the side effects of 5-FU, we have designed and synthesized delocalized lipophilic cations (DLCs) as a vehicle for the delivery of 5-FU. DLCs accumulate selectively in the mitochondria of cancer cells because of the high mitochondrial transmembrane potential (ΔΨm). Many DLCs exhibited anti-cancer efficacy and were explored as potential anti-cancer drugs based on their selective accumulation in the mitochondria of cancer cells. F16, the DLC we used as a vehicle, is a small molecule that selectively inhibits tumor cell growth and dissipates mitochondrial membrane potential. The binding of the conjugate F16-5-FU to bovine serum albumin (BSA) was investigated using spectroscopic and molecular modeling approaches. Fluorescence quenching constants were determined using the Stern-Volmer equation to provide a measure of the binding affinity between F16-5-FU and BSA. The activation energy of the interaction between F16-5-FU and BSA was calculated and the unusually high value was discussed in terms of the special structural block indicated by the molecular modeling approach. Molecular modeling showed that F16-5-FU binds to human serum albumin in site II, which is consistent with the results of site-competitive replacement experiments. It is suggested that hydrophobic and polar forces played important roles in the binding reaction, in accordance with the results of thermodynamic experiments.


Direct effect of rituximab in B-cell-derived lymphoid neoplasias: mechanism, regulation, and perspectives.
The anti-CD20 monoclonal antibody rituximab is the backbone of treatment for the B-cell malignancies non-Hodgkin lymphoma and chronic lymphocytic leukemia. However, there is a wide variability in response to rituximab treatment, and some patients are refractory to current standard therapies. Rituximab kills B cells by multiple mechanisms of action, including complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity, which are immune-mediated mechanisms, as well as by direct effects on cell signaling pathways and cell membranes following CD20 binding. A large number of events that are affected by rituximab binding have been identified, including lipid raft modifications, kinase and caspase activation, and effects on transcription factors and apoptotic/antiapoptotic molecules. Studies on cell lines and isolated tumor cells have shown that by targeting these pathways, it may be possible to increase or decrease susceptibility to rituximab cell killing. An increased understanding of the direct effects of rituximab may therefore aid in the design of new, rational combinations to improve the outcome of CD20-based therapy for patients who currently have suboptimal outcome following standard treatments.


The Importance and Place of Adenotonsillectomy in Syndromic Children.
Upper airway obstruction and sleeping disorders are important issues in syndromic children, including mouth, lower-upper jaw, or all facial abnormalities. Tonsillectomy and/or adenoidectomy is required because of systemic problems and upper airway obstruction that increase the existing systemic problems, except those anomalies. However, tonsillectomy and/or adenoidectomy are mostly avoided because of the tendency to cause both intubation/perioperative systemic problems and respiratory complications in the postoperative period and in delays in the oral intake. However, these surgeries are sometimes required. In this context, we present our experience related with performing tonsillectomy and/or adenoidectomy in syndromic children admitted to our hospital. We retrospectively examined the data on tonsillectomy and/or adenoidectomy performed in syndromic patients in our clinic between 2001 and 2011. We did not observe any postoperative complications in adenoidectomy and/or tonsillectomy performed by the same surgeon in 14 syndromic cases. It should be noted that respiratory problems may arise from many different anatomical regions in syndromic patients. Therefore, surgery should be performed taking into consideration all of these factors in these patients. These patients must be hospitalized in the postoperative period.


Configuration of Ten Light-Harvesting Chlorophyll a/b Complex I Subunits in Chlamydomonas reinhardtii Photosystem I.
In plants, the photosystem I (PSI) core complex stably associates with its light-harvesting chlorophyll a/b complex I (LHCI) to form the PSI-LHCI supercomplex. The vascular plant PSI core complex associates with four distinct LHCI subunits, whereas that of the green alga Chlamydomonas reinhardtii binds nine distinct LHCI subunits (LHCA1-LHCA9). The stoichiometry and configuration of these LHCI subunits in the PSI-LHCI supercomplex of C. reinhardtii remain controversial. Here, we determined the stoichiometry of the nine distinct LHCI subunits relative to PSI subunits through uniform labeling of total proteins using 14C. We separated the nine LHCI polypeptides by three different sodium dodecyl sulfate-polyacrylamide gel electrophoresis systems. Our data revealed that the PSI-LHCI supercomplex contains two LHCA1 proteins and one of each of the other eight LHCI subunits. Subsequently, we identified their cross-linked products by immunodetection and mass spectrometry to determine the configuration of the 10 LHCI subunits within the PSI-LHCI supercomplex. Furthermore, analyses of PSI-LHCI complexes isolated from ΔLHCA2 and ΔLHCA5 mutants and oligomeric LHCI from a PSI-deficient (ΔpsaA/B) mutant provided supporting evidence for the LHCI subunit configuration. In conclusion, eight LHCI subunits bind to the PSI core at the site of PSAF subunit in two layers: LHCA1-LHCA8-LHCA7-LHCA3 from PSAG to PSAK, in the inner layer, and LHCA1-LHCA4-LHCA6-LHCA5 in the outer layer. The other two LHCI subunits, LHCA2 and LHCA9, bind PSAB between PSAG and PSAH, PSAG-LHCA9-LHCA2-PSAH. Our study provides new insights into the LHCI configuration linked to the PSI core.


Preoperative staging of rectal cancer: accuracy of 3-Tesla magnetic resonance imaging.
The purpose of this study was to evaluate the accuracy of 3-Tesla magnetic resonance imaging (MRI) for the preoperative staging of rectal cancer. Thirty-five patients with a primary rectal cancer who underwent preoperative 3-T MRI using a phased-array coil and had a surgical resection were enrolled in the study group. Preoperatively, three experienced radiologists independently assessed the T and N staging. A confidence level scoring system was used to determine if there was any perirectal invasion, and receiver operating characteristic (ROC) curves were generated. The interobserver agreement was estimated using kappa statistics. The overall accuracy rate of T staging for rectal cancer was 92%. The diagnostic accuracy was 97% for T1, 89% for T2 and 91% for T3, respectively. The predictive accuracy for perirectal invasion by the three observers was high (Az>0.92). The interobserver agreement for T staging was moderate to substantial. The overall sensitivity, specificity, and accuracy for the detection of mesorectal nodal metastases were 80%, 98%, and 95%, respectively. In conclusion, preoperative 3-T MRI using a phase-array coil accurately indicates the depth of tumor invasion for rectal cancer with a low variability.


Relationship between serum acetaminophen concentration and N-acetylcysteine-induced adverse drug reactions.
Intravenous N-acetylcysteine is usually regarded as a safe antidote. However, during the infusion of the loading dose, different types of adverse drug reactions (ADR) may occur. The objective of this study was to investigate the relation between the incidence of different types of ADR and serum acetaminophen concentration in patients presenting to the hospital with acetaminophen overdose. This is a retrospective study of patients admitted to the hospital for acute acetaminophen overdose over a period of 5 years (1 January 2004 to 31 December 2008). Parametric and non-parametric tests were used to test differences between groups depending on the normality of the data. SPSS 15 was used for data analysis. Of 305 patients with acetaminophen overdose, 146 (47.9%) were treated with intravenous N-acetylcysteine and 139 (45.6%) were included in this study. Different types of ADR were observed in 94 (67.6%) patients. Low serum acetaminophen concentrations were significantly associated with cutaneous anaphylactoid reactions but not other types of ADR. Low serum acetaminophen concentration was significantly associated with flushing (p < 0.001), rash (p < 0.001) and pruritus (p < 0.001). However, there were no significant differences in serum acetaminophen concentrations between patients with and without the following ADR: gastrointestinal reactions (p = 0.77), respiratory reactions (p = 0.96), central nervous reactions (p = 0.82) and cardiovascular reactions (p = 0.37). In conclusion, low serum acetaminophen concentrations were associated with higher cutaneous anaphylactoid reactions. Such high serum acetaminophen concentrations may be protective against N-acetylcysteine-induced cutaneous ADR.


Validity of the brief pain inventory for use in documenting the outcomes of patients with noncancer pain.
The Brief Pain Inventory (BPI) is a short, self-administered questionnaire that was developed for use in cancer patients. While most empirical research with the BPI has been in pain of that etiology, the questionnaire is increasingly evident in published studies of patients with non-cancer pain. The current research addresses the need for formal evaluation of the reliability and validity of the BPI for use in non-cancer pain patients. Approximately 250 patients with arthritis or low back pain (LBP) self-administered a number of generic and condition-specific health status measures (including the BPI) in the clinic of their primary care provider at 2 time points: the initial clinic visit and the first visit following treatment. The reliability of BPI data collected from non-cancer pain patients was comparable to that reported in the literature for cancer patients and sufficient for group-level analyses (coefficient alphas were greater than 0.70). The factor structure of the BPI was replicated in this sample and the relationship of the BPI to generic measures of pain was strong. The BPI exhibited similar relationships to general and condition-specific measures of health as did a generic pain scale (SF-36 Bodily Pain). Finally, the BPI discriminated among levels of condition severity and was sensitive to change in condition over time in arthritis and LBP patients. Results support the validity of the BPI as a measure of pain in patients without cancer and, in particular, as a measure of pain for arthritis and LBP patients.


Functional characteristics of an endophyte community colonizing rice roots as revealed by metagenomic analysis.
Roots are the primary site of interaction between plants and microorganisms. To meet food demands in changing climates, improved yields and stress resistance are increasingly important, stimulating efforts to identify factors that affect plant productivity. The role of bacterial endophytes that reside inside plants remains largely unexplored, because analysis of their specific functions is impeded by difficulties in cultivating most prokaryotes. Here, we present the first metagenomic approach to analyze an endophytic bacterial community resident inside roots of rice, one of the most important staple foods. Metagenome sequences were obtained from endophyte cells extracted from roots of field-grown plants. Putative functions were deduced from protein domains or similarity analyses of protein-encoding gene fragments, and allowed insights into the capacities of endophyte cells. This allowed us to predict traits and metabolic processes important for the endophytic lifestyle, suggesting that the endorhizosphere is an exclusive microhabitat requiring numerous adaptations. Prominent features included flagella, plant-polymer-degrading enzymes, protein secretion systems, iron acquisition and storage, quorum sensing, and detoxification of reactive oxygen species. Surprisingly, endophytes might be involved in the entire nitrogen cycle, as protein domains involved in N(2)-fixation, denitrification, and nitrification were detected and selected genes expressed. Our data suggest a high potential of the endophyte community for plant-growth promotion, improvement of plant stress resistance, biocontrol against pathogens, and bioremediation, regardless of their culturability.


Generation of random numbers on graphics processors: forced indentation in silico of the bacteriophage HK97.
The use of graphics processing units (GPUs) in simulation applications offers a significant speed gain as compared to computations on central processing units (CPUs). Many simulation methods require a large number of independent random variables generated at each step. We present two approaches for implementation of random number generators (RNGs) on a GPU. In the one-RNG-per-thread approach, one RNG produces a stream of random numbers in each thread of execution, whereas the one-RNG-for-all-threads method builds on the ability of different threads to communicate, thus, sharing random seeds across an entire GPU device. We used these approaches to implement Ran2, Hybrid Taus, and Lagged Fibonacci algorithms on a GPU. We profiled the performance of these generators in terms of the computational time, memory usage, and the speedup factor (CPU time/GPU time). These generators have been incorporated into the program for Langevin simulations of biomolecules fully implemented on the GPU. The ∼250-fold computational speedup on the GPU allowed us to carry out single-molecule dynamic force measurements in silico to explore the mechanical properties of the bacteriophage HK97 in the experimental subsecond time scale. We found that the nanomechanical response of HK97 depends on the conditions of force application, including the rate of change and geometry of the mechanical perturbation. Hence, using the GPU-based implementation of RNGs, presented here, in conjunction with Langevin simulations, makes it possible to directly compare the results of dynamic force measurements in vitro and in silico.


Health promotion and surveillance: the establishment of an IUHPE global working group.
Following a series of international meetings on behavioral monitoring and surveillance, in 2007 the Italian Ministry of Health (Ministerio della Salute) and the Institute for Health (Istituto Superiore della Sanità) hosted the 5(th) International Conference on Behavioral Risk Factor Surveillance (BRFS) in Rome. A key focus of the conference was on how current surveillance systems could be applied to the field of health promotion, particularly in building the evidence base for health promotion practice. As a result of these discussions, the World Alliance for Risk Factor Surveillance (WARFS), an IUHPE Global Working Group, was formed to work toward providing knowledge and expertise in surveillance as a tool for advancing health promotion. For those IUHPE members interested in participation, this article provides an overview on the strategic direction of WARFS and the newly formed sub-working groups.


Impact of increasing the availability of healthier vs. less-healthy food on food selection: a randomised laboratory experiment.
Environmental cues shape behaviour, but few studies compare the impact of targeting healthier vs. less-healthy cues. One online study suggested greater impact on selection from increasing the number of less-healthy (vs. healthier) snacks. The current study aimed to: (1) extend the previous study by using physically-present snacks for immediate consumption; (2) explore responsiveness by socio-economic position; (3) investigate possible mediators (response inhibition, food appeal) of any socio-economic differences in selection. In a between-subjects laboratory experiment UK adults (n = 417) were randomised according to their ID number (without blinding) to one of three ranges of options: Two healthier, two less-healthy ["Equal"] (n = 136); Six healthier, two less-healthy ["Increased Healthier"] (n = 143); Two healthier, six less-healthy ["Increased Less-Healthy"] (n = 138). Participants completed measures of response inhibition and food appeal, and selected a snack for immediate consumption from their allocated range. The primary outcome was selection of a healthier (over less-healthy) snack. The odds of selecting a less-healthy snack were 2.9 times higher (95%CIs:1.7,5.1) in the Increased Less-Healthy condition compared to the Equal condition. The odds of selecting a healthier snack were 2.5 times higher (95%CIs:1.5,4.1) in the Increased Healthier (vs. Equal) condition. There was no significant difference in the size of these effects (- 0.2; 95%CIs:-1.1,0.7). Findings were inconclusive with regard to interactions by education, but the direction of effects was consistent with potentially larger impact of the Increased Healthier condition on selection for higher-educated participants, and potentially larger impact of the Increased Less-Healthy condition for less-educated participants. A greater impact from increasing the number of less-healthy (over healthier) foods was not replicated when selecting snacks for immediate consumption: both increased selections of the targeted foods with no evidence of a difference in effectiveness. The observed pattern of results suggested possible differential impact by education, albeit not statistically significant. If replicated in larger studies, this could suggest that removing less-healthy options has the potential to reduce health inequalities due to unhealthier diets. Conversely, adding healthier options could have the potential to increase these inequalities. ISRCTN: ISRCTN34626166 ; 11/06/2018; Retrospectively registered.


Heterogeneity of D2 dopamine receptors in different brain regions.
The binding of [3H]spiperone has been examined in membranes derived from different regions of bovine brain. In caudate nucleus, nucleus accumbens, olfactory tubercle and putamen binding is to D2 dopamine and 5HT2 serotonin receptors, whereas in cingulate cortex only serotonin 5HT2 receptor binding can be detected. D2 dopamine receptors were examined in detail in caudate nucleus, olfactory tubercle and putamen using [3H]spiperone binding in the presence of 0.3 microM-mianserin (to block 5HT2 serotonin receptors). No evidence for heterogeneity among D2 dopamine receptors either between brain regions or within a brain region was found from the displacements of [3H]spiperone binding by a range of antagonists, including dibenzazepines and substituted benzamides. Regulation of agonist binding by guanine nucleotides did, however, differ between regions. In caudate nucleus a population of agonist binding sites appeared resistant to guanine nucleotide regulation, whereas this was not the case in olfactory tubercle and putamen.


Corticosteroids improve short-term survival in patients with severe alcoholic hepatitis: meta-analysis of individual patient data.
A meta-analysis was performed using individual patient data from the five most recent randomised controlled trials (RCTs) which evaluated corticosteroids in severe alcoholic hepatitis (Maddrey discriminant function (DF) ≥ 32 or encephalopathy). This approach overcomes limitations associated with the use of literature data and improves the relevance of the study and estimates of effect size. To compare 28-day survival between corticosteroid- and non-corticosteroid-treated patients and to analyse the response to treatment using the Lille model. Individual patient data were obtained from five RCTs comparing corticosteroid treatment with placebo (n=3), enteral nutrition (n=1) or an antioxidant cocktail (n=1). 221 patients allocated to corticosteroid treatment and 197 allocated to non-corticosteroid treatment were analysed. The two groups were similar at baseline. 28-day survival was higher in corticosteroid-treated patients than in non-corticosteroid-treated patients (79.97±2.8% vs 65.7±3.4%, p=0.0005). In multivariate analysis, corticosteroids (p=0.005), DF (p=0.006), leucocytes (p=0.004), Lille score (p<0.00001) and encephalopathy (p=0.003) were independently predictive of 28-day survival. A subgroup analysis was performed according to the percentile distribution of the Lille score. Patients were classified as complete responders (Lille score ≤ 0.16; ≤ 35th percentile), partial responders (Lille score 0.16-0.56; 35th-70th percentile) and null responders (Lille ≥ 0.56; ≥ 70th percentile). 28-day survival was strongly associated with these groupings (91.1±2.7% vs 79.4±3.8% vs 53.3±5.1%, p<0.0001). Corticosteroids had a significant effect on 28-day survival in complete responders (HR 0.18, p=0.006) and in partial responders (HR 0.38, p=0.04) but not in null responders. Analysis of individual data from five RCTs showed that corticosteroids significantly improve 28-day survival in patients with severe alcoholic hepatitis. The survival benefit is mainly observed in patients classified as responders by the Lille model.


Methadone safety: a clinical practice guideline from the American Pain Society and College on Problems of Drug Dependence, in collaboration with the Heart Rhythm Society.
Methadone is used for the treatment of opioid addiction and for treatment of chronic pain. The safety of methadone has been called into question by data indicating a large increase in the number of methadone-associated overdose deaths in recent years that has occurred in parallel with a dramatic rise in the use of methadone for chronic pain. The American Pain Society and the College on Problems of Drug Dependence, in collaboration with the Heart Rhythm Society, commissioned an interdisciplinary expert panel to develop a clinical practice guideline on safer prescribing of methadone for treatment of opioid addiction and chronic pain. As part of the guideline development process, the American Pain Society commissioned a systematic review of various aspects related to safety of methadone. After a review of the available evidence, the expert panel concluded that measures can be taken to promote safer use of methadone. Specific recommendations include the need to educate and counsel patients on methadone safety, use of electrocardiography to identify persons at greater risk for methadone-associated arrhythmia, use of alternative opioids in patients at high risk of complications related to corrected electrocardiographic QTc interval prolongation, careful dose initiation and titration of methadone, and diligent monitoring and follow-up. Although these guidelines are based on a systematic review, the panel identified numerous research gaps, most recommendations were based on low-quality evidence, and no recommendations were based on high-quality evidence. This guideline, based on a systematic review of the evidence on methadone safety, provides recommendations developed by a multidisciplinary expert panel. Safe use of methadone requires clinical skills and knowledge in use of methadone to mitigate potential risks, including serious risks related to risk of overdose and cardiac arrhythmias.


What is quality, and can we define it in lung cancer?-the case for quality improvement.
Decades worth of advances in diagnostics and therapeutics are associated with only marginal improvements in survival among lung cancer patients. An obvious explanation is late stage at presentation, but gaps in the quality of care may be another reason for stifled improvements in survival rates. A framework for quality put forth by Avedis Donabedian consists of measuring structures-of-care, processes, and outcomes. Using this approach to explore for potential quality gaps, there is evidence of inexplicable variability in outcomes across patients and hospitals; variation in outcomes across differing provider types (structures-of-care); and variation in approaches to staging (processes-of-care). However, this research has limitations and incontrovertible evidence of quality gaps is challenging to obtain. Other challenges to defining quality include scientific and clinical uncertainty among providers and the fact that quality is a multi-dimensional construct that cannot be measured by a single metric. Nonetheless, two facts compel us to pursue quality improvement: (I) both empirically and anecdotally, actual care falls short of expected care; and (II) evidence of potential quality gaps is not ignorable primarily on ethical grounds.


Isolation-by-environment as a driver of genetic differentiation among populations of the only broad-leaved evergreen shrub Ammopiptanthus mongolicus in Asian temperate deserts.
Whether the effect of migration-selection-drift equilibrium on population structure is governed by spatial or environmental differences is usually elucidated by isolation-by-distance (IBD), isolation-by-environment (IBE), and isolation-by-resistance (IBR) tests. The population structure of Ammopiptanthus mongolicus, a broad-leaved evergreen psammophyte in eastern Central Asia, was previously thought to follow an isolation by distance pattern. However, recent studies have emphasized the effects of environmental factors on its growth and distribution, suggesting an important influence of local adaptation on the genetic structure of the species. Using inter-simple sequence repeat (ISSR) markers, we verified the previously inferred low intra-population variation and high inter-population differentiation. However, in contrast to previous studies, the results of partial Mantel tests and a maximum likelihood population effects mixed model (MLPE) suggested that local climate differences, rather than geographic distances or resistance distances, are the main factor affecting population differentiation. Further analysis with removal of multicollinear climatic variables and univariate MLPE found that summer and winter precipitation were crucial for shaping the current population genetic structure. Since local precipitation is related to the regeneration, colonization, and overwintering survival of A. mongolicus, its influence on demographic change may explain its effect on the population genetic structure. In addition, precipitation is related to terrain despite westward decreases, which explains the independence of genetic difference and geographic distance. The identified role of IBE suggests that collecting germplasm resources from genetically differentiated populations could be a more effective strategy to preserve the overall genetic diversity of the species than the establishment of corridors to enhance gene flow among populations.


Daily low-dose carboplatin as a radiation sensitizer for newly diagnosed malignant glioma.
Surgical resection followed by local field radiotherapy is currently our most effective approach to treatment for most patients with malignant glioma. Carboplatin chemotherapy has direct cytotoxic effects on glioma cells and acts as a radiation sensitizer to enhance cell killing. Its demonstrated efficacy as a sensitizer in other solid tumors led to this clinical trial of carboplatin as a radiation sensitizer in the treatment of newly diagnosed glioblastoma multiforme (GBM) and anaplastic astrocytoma (AA). Fourteen patients (nine GBM and five AA) were treated with daily low-dose carboplatin 25 mg/m2 intravenously within 2 h of their fractionated radiotherapy to a total dose of 600 mg/m2. No significant toxicities attributable to this combined therapy were observed. All patients have progressed, with median time to progression of 16 weeks. Eleven patients have died, with median survival of 38 weeks for the entire cohort. Although this regimen appeared safe, there was no benefit in survival time compared to historical patients treated with radiotherapy. The limitations and future potential for the strategy of radiation sensitization are discussed.


Automated detection of exudates in colored retinal images for diagnosis of diabetic retinopathy.
Medical image analysis is a very popular research area these days in which digital images are analyzed for the diagnosis and screening of different medical problems. Diabetic retinopathy (DR) is an eye disease caused by the increase of insulin in blood and may cause blindness. An automated system for early detection of DR can save a patient's vision and can also help the ophthalmologists in screening of DR. The background or nonproliferative DR contains four types of lesions, i.e., microaneurysms, hemorrhages, hard exudates, and soft exudates. This paper presents a method for detection and classification of exudates in colored retinal images. We present a novel technique that uses filter banks to extract the candidate regions for possible exudates. It eliminates the spurious exudate regions by removing the optic disc region. Then it applies a Bayesian classifier as a combination of Gaussian functions to detect exudate and nonexudate regions. The proposed system is evaluated and tested on publicly available retinal image databases using performance parameters such as sensitivity, specificity, and accuracy. We further compare our system with already proposed and published methods to show the validity of the proposed system.


Covalent protein display on Hepatitis B core-like particles in plants through the in vivo use of the SpyTag/SpyCatcher system.
Virus-like particles (VLPs) can be used as nano-carriers and antigen-display systems in vaccine development and therapeutic applications. Conjugation of peptides or whole proteins to VLPs can be achieved using different methods such as the SpyTag/SpyCatcher system. Here we investigate the conjugation of tandem Hepatitis B core (tHBcAg) VLPs and the model antigen GFP in vivo in Nicotiana benthamiana. We show that tHBcAg VLPs could be successfully conjugated with GFP in the cytosol and ER without altering VLP formation or GFP fluorescence. Conjugation in the cytosol was more efficient when SpyCatcher was displayed on tHBcAg VLPs instead of being fused to GFP. This effect was even more obvious in the ER, showing that it is optimal to display SpyCatcher on the tHBcAg VLPs and SpyTag on the binding partner. To test transferability of the GFP results to other antigens, we successfully conjugated tHBcAg VLPs to the HIV capsid protein P24 in the cytosol. This work presents an efficient strategy which can lead to time and cost saving post-translational, covalent conjugation of recombinant proteins in plants.


Dependence of the Ce(iii)/Ce(iv) ratio on intracellular localization in ceria nanoparticles internalized by human cells.
CeO2 nanoparticles (CNPs) have been investigated as promising antioxidant agents with significant activity in the therapy of diseases involving free radicals or oxidative stress. However, the exact mechanism responsible for CNP activity has not been completely elucidated. In particular, in situ evidence of modification of the oxidative state of CNPs in human cells and their evolution during cell internalization and subsequent intracellular distribution has never been presented. In this study we investigated modification of the Ce(iii)/Ce(iv) ratio following internalization in human cells by X-ray absorption near edge spectroscopy (XANES). From this analysis on cell pellets, we observed that CNPs incubated for 24 h showed a significant increase in Ce(iii). By coupling on individual cells synchrotron micro-X-ray fluorescence (μXRF) with micro-XANES (μXANES) we demonstrated that the Ce(iii)/Ce(iv) ratio is also dependent on CNP intracellular localization. The regions with the highest CNP concentrations, suggested to be endolysosomes by transmission electron microscopy, were characterized by Ce atoms in the Ce(iv) oxidation state, while a higher Ce(iii) content was observed in regions surrounding these areas. These observations suggest that the interaction of CNPs with cells involves a complex mechanism in which different cellular areas play different roles.


Learning about COVID-19-related stigma, quarantine and isolation experiences in Finland.
The COVID-19 pandemic has intensely changed the everyday lives of people worldwide. This study explores the forms and outcomes of coronavirus and COVID-19-related social stigma and the experiences of people who were home quarantined or isolated in Finland during the spring 2020. The findings of this study can be used to improve support for those quarantined or isolated and to develop strategies to reduce the stigma associated with coronavirus and COVID-19. The study is based on qualitative one-to-one interviews with households with at least two members and at least one PCR confirmed COVID-19 case. Recruitment took place via website or SMS messages sent to PCR confirmed cases in the capital area of Helsinki. Sampling was based on maximum variation to acquire different types of respondents. The framework of health stigma was used to develop question guides and analyze stigma. Quarantine and isolation experiences were explored through open-ended questions. The analysis was based on thematic analysis. The study included 64 participants from 24 households. Perceived stigma among respondents was driven by fear and blame for infection, and it manifested in various ways leading to a reluctance to disclose their coronavirus status to others. Self-stigma developed from conflicting information and advice about coronavirus and COVID-19 led to difficulties interacting with others outside of the house and reluctance to meet people after quarantine and isolation. Quarantine and isolation experiences included uncertainty, health concerns, and boredom. Communication with others in similar situations was perceived vital, whereas discussions with family members about worries and fears related to coronavirus and COVID-19 was not preferred. This study shed light on the lives of those quarantined or isolated at home and provided a set of operational recommendations to minimize coronavirus and COVID-19-associated stigma and to reduce challenges faced by those in quarantine or isolation.


Fracture Healing Monitoring by Impact Tests: Single Case Study of a Fractured Tibia With External Fixator.
The correct evaluation of the healing process is important to define proper times of fixator dynamization and removal, avoiding refractures. Unfortunately, a quantitative healing assessment is not yet available in clinical practice. The aim of the paper is to prove the feasibility of the mechanical vibration method to assess bone healing in fractures treated with external fixation, in in vivo conditions. The case study was a patient with a tibial fracture treated with a monoaxial fixator. The healing process was monitored for three months through a series of five impact tests. The pins screwed into the bone were used both to excite and measure vibrations. Fracture healing was quantitatively assessed by estimating the resonant frequencies of the leg. The first frequency increased of about 4% per week during the observation period. After the hard callus formation (13 week), also other frequencies increased within the range 1%-6% per week. X-ray observations confirmed the healing progress and proved the method potentiality. In addition, the vibratory response of the leg after fixator removal was evaluated and resulted characterized by five modes in the bandwidth 0-1000 Hz. The results suggest that the vibratory response of a fractured bone treated with external fixation can be a promising indicator for quantitative healing monitoring. The mechanical vibration method could be helpful for reducing X-ray exposure of patients and could be performed more frequently, as desirable for obtaining more attentive monitoring.


Using DNA duplex stability information for transcription factor binding site discovery.
Transcription factor (TF) binding site discovery is an important step in understanding transcriptional regulation. Many computational tools have already been developed, but their success in detecting TF motifs is still limited. We believe one of the main reasons for the low accuracy of current methods is that they do not take into account the structural aspects of TF-DNA interaction. We have previously shown that knowledge about the structural class of the TF and information about nucleosome occupancy can be used to improve motif discovery. Here, we demonstrate the benefits of using information about the DNA double-helical stability for motif discovery. We notice that, in general, the energy needed to destabilize the DNA double helix is higher at TF binding sites than at random DNA sites. We use this information to derive informative positional priors that we incorporate into a motif finding algorithm. When applied to yeast ChIP-chip data, the new informative priors improve the performance of the motif finder significantly when compared to priors that do not use the energetic stability information.


Structural and functional characterization of the PDZ domain of the human phosphatase PTPN3 and its interaction with the human papillomavirus E6 oncoprotein.
The human protein tyrosine phosphatase non-receptor type 3 (PTPN3) is a PDZ (PSD-95/Dlg/ZO-1) domain-containing phosphatase with a tumor-suppressive or a tumor-promoting role in many cancers. Interestingly, the high-risk genital human papillomavirus (HPV) types 16 and 18 target the PDZ domain of PTPN3. The presence of a PDZ binding motif (PBM) on E6 confers interaction with a number of different cellular PDZ domain-containing proteins and is a marker of high oncogenic potential. Here, we report the molecular basis of interaction between the PDZ domain of PTPN3 and the PBM of the HPV E6 protein. We combined biophysical, NMR and X-ray experiments to investigate the structural and functional properties of the PDZ domain of PTPN3. We showed that the C-terminal sequences from viral proteins encompassing a PBM interact with PTPN3-PDZ with similar affinities to the endogenous PTPN3 ligand MAP kinase p38γ. PBM binding stabilizes the PDZ domain of PTPN3. We solved the X-ray structure of the PDZ domain of PTPN3 in complex with the PBM of the HPV E6 protein. The crystal structure and the NMR chemical shift mapping of the PTPN3-PDZ/peptide complex allowed us to pinpoint the main structural determinants of recognition of the C-terminal sequence of the E6 protein and the long-range perturbations induced upon PBM binding.


Preoperative distraction in children: hand-held videogames vs clown therapy.
Anxiety in children undergoing surgery is characterized by feelings of tension, apprehension, nervousness and fear which may manifest differently. Postoperative behavioural changes such as nocturnal enuresis, feeding disorders, apathy, and sleep disturbances may stem from postoperative anxiety. Some Authors pointed out that over 60% of children undergoing surgery are prone to developing behavioural alterations 2 weeks after surgery. Variables such as age, temperament and anxiety both in children and parents are considered predictors of such changes.1 Studies were published describing how psycho-behavioural interventions based on play, learning and entertainment in preparing children for surgery, may reduce preoperative anxiety. Clown-therapy is applied in the most important paediatric facilities and has proved to diminish children's emotional distress and sufferance, as well as consumption of both analgesics and sedatives and to facilitate the achievement of therapeutic goals. The aim of our study was to evaluate the efficacy of clown-therapy during the child's hospital stay, with a view to optimizing treatment and care, preventing behavioural alterations and enhancing the child's overall life quality.


Real-Time Lane Region Detection Using a Combination of Geometrical and Image Features.
Over the past few decades, pavement markings have played a key role in intelligent vehicle applications such as guidance, navigation, and control. However, there are still serious issues facing the problem of lane marking detection. For example, problems include excessive processing time and false detection due to similarities in color and edges between traffic signs (channeling lines, stop lines, crosswalk, arrows, etc.). This paper proposes a strategy to extract the lane marking information taking into consideration its features such as color, edge, and width, as well as the vehicle speed. Firstly, defining the region of interest is a critical task to achieve real-time performance. In this sense, the region of interest is dependent on vehicle speed. Secondly, the lane markings are detected by using a hybrid color-edge feature method along with a probabilistic method, based on distance-color dependence and a hierarchical fitting model. Thirdly, the following lane marking information is extracted: the number of lane markings to both sides of the vehicle, the respective fitting model, and the centroid information of the lane. Using these parameters, the region is computed by using a road geometric model. To evaluate the proposed method, a set of consecutive frames was used in order to validate the performance.


Development of the Polypay breed of sheep.
Development of the Polypay breed was undertaken to combine into a composite breed the potential for greatly increased reproductive capacity along with desirable growth rate and carcass quality. Four breeds were selected for the foundation of the new breed--the Rambouillet and the Targhee for hardiness, large body size, long breeding season, herding instinct and fleece characteristics; the Dorset for carcass quality, milking ability and long breeding season and the Finnsheep for early puberty, early postpartum fertility and high lambing rate. Dorset X Targhee and Finnsheep X Rambouillet matings were first made in 1968, and reciprocal Dorset-Targhee X Finn-Rambouillet matings were initiated in 1969. Beginning in 1970, the respective two-breed crosses and the four-breed cross (Polypay) were each mated inter se and selected, along with straightbred Rambouillets and Targhees, for lamb production when given two opportunities to lamb/year. Initial comparisons among straightbreds and inter se mated groups showed few important differences in meat-type body conformation, body condition or growth rate, but superior annual reproductive performance by Polypays. Fertility of Polypays at 1 yr of age was high and comparable to that of the Finn-Rambouillets. Response of Polypays to twice-a-year lambing was superior to responses of Rambouillets. Targhees, Dorset-Targhees or Finn-Rambouillets. The 1974 to 1975 annual production of young Polypays was about 13% more lambs weaned than from Finn-Rambouillets and 18% more weight of lamb weaned than from Dorset-Targhees, the best of the other groups for these traits. The current (1979 to 1981) reproductive performance of Polypay ewes selected for high once-a-year lambing rate under typical range management conditions is very competitive with that of 1/2 Finn crossbreds. Annual production of mature Polypays on the twice-a-year lambing schedule was 1.78 lambs weaned and 58.6 kg of lamb weaned/ewe put into fall breeding.