Current cigarette smoking among adults--United States, 2005-2013.
Tobacco use is the leading cause of preventable disease and death in the United States, resulting in more than 480,000 premature deaths and $289 billion in direct health care expenditures and productivity losses each year. Despite progress over the past several decades, millions of adults still smoke cigarettes, the most commonly used tobacco product in the United States. To assess progress made toward the Healthy People 2020 target of reducing the proportion of U.S. adults who smoke cigarettes to ≤12.0% (objective TU-1.1), CDC used data from the 2013 National Health Interview Survey (NHIS) to provide updated national estimates of cigarette smoking prevalence among adults aged ≥18 years. Additionally, for the first time, estimates of cigarette smoking prevalence were assessed among lesbian, gay, or bisexual persons (LGB) using NHIS data. The proportion of U.S. adults who smoke cigarettes declined from 20.9% in 2005 to 17.8% in 2013, and the proportion of daily smokers declined from 16.9% to 13.7%. Among daily cigarette smokers, the proportion who smoked 20-29 cigarettes per day (CPD) declined from 34.9% to 29.3%, and the proportion who smoked ≥30 CPD declined from 12.7% to 7.1%. However, cigarette smoking remains particularly high among certain groups, including adults who are male, younger, multiracial or American Indian/Alaska Native, have less education, live below the federal poverty level, live in the South or Midwest, have a disability/limitation, or who are LGB. Proven population-based interventions, including tobacco price increases, comprehensive smoke-free policies in worksites and public places, high-impact anti-tobacco mass media campaigns, and easy access to smoking cessation assistance, are critical to reducing cigarette smoking and smoking-related disease and death among U.S. adults, particularly among subpopulations with the greatest burden.


The association between medical costs and participation in the vitality health promotion program among 948,974 members of a South African health insurance company.
Examine the association between the levels of participation in an incentive-based health promotion program (Vitality) and inpatient medical claims among members of a major health insurer. A 1-year, cross-sectional, correlational analyses of engagement with a health promotion program and hospital claims experience (admissions costs, days in hospital, and admission rate) of members of a national private health insurer. Adult members of South Africa's largest national private health insurer, Discovery Health. Insured members were also eligible for voluntary membership in an insurance-linked incentivized health promotion program, Vitality. The study sample included 948,974 adult members of the Discovery Health plan for the year 2006. Of these, 591,134 (62.3%) were also members of the Vitality health promotion program. The study sample was grouped based on registration and the level of engagement with the Vitality health promotion program into the following: not registered (37.5%), registered but not engaged with any health promotion activity (21.9%), low engagement (30.9%), and high engagement (9.5%). High engagement was defined a priori by the accumulation of an arbitrary number of points on the Vitality program, allocated against specific activities (knowledge, fitness-related activities, assessment and screening, and healthy choices). Hospital admission costs, the number of days in hospital, and hospital admission rates were compared among highly engaged members and those members who were not enrolled in the program, nonengaged, and lowly engaged. Data were normalized for age, gender, plan type, and chronic disease status. Highly engaged members had lower costs per patient, shorter stays in hospital, and fewer admissions compared with other groups (p < .001). Low or no engagement was not associated with lower hospital costs. Admission rates were also 7.4% lower for cardiovascular disease, 13.2% lower for cancers, and 20.7% lower for endocrine and metabolic diseases in the highly engaged group compared with any of the other groups (p < .01). Engagement in an incentive-based wellness program, offered by a health insurer, was associated with lower health care costs.


MHC class I molecules are an essential cell surface component involved in Theileria parva sporozoite binding to bovine lymphocytes.
The major histocompatibility complex (MHC) class I molecules are ubiquitous cell surface molecules involved in the cell-mediated immune response. We show here, using a number of different, independent approaches, that these proteins are an essential component of the host cell surface receptor involved in Theileria parva sporozoite invasion. Monoclonal antibodies (mAbs) reactive with common determinants on MHC class I molecules and with beta-2 microglobulin inhibited sporozoite entry by specifically preventing the initial binding event. However, in experiments using lymphocytes from heterozygous cattle in which at least four MHC class I gene products are expressed, mAbs which reacted with only one of these products did not inhibit entry. Using a series of bovine deletion mutant cell lines from which one or both MHC class I haplotypes had been lost, sporozoite binding and entry clearly correlated with the level of class I surface expression. While the level of sporozoite entry into cells in which one of the MHC class I haplotypes was lost was only slightly lower than into the parent cells, in a double deletion cell line having less than 5% of the class I expression of the parent cells the level of infection was only 4.3% of that into the parent cells. Furthermore, sporozoite entry into cells from a spontaneously arising mutant cell line exhibiting low levels of class I expression was correspondingly low. Treatment of lymphocytes with IL-2 produced a significant increase in host cell susceptibility and sporozoite entry and this increase correlated with either an increase in the number of target molecules per host cell, or in the binding of bovine MHC class I molecules to the mAbs. In particular, a significant increase in the level of reactivity with mAb W6/32 was observed. Lastly, we show that parasite entry can be competitively inhibited with an isolated sporozoite surface protein, p67. However, p67 binds weakly to lymphocyte surface molecules and initial attempts to use p67 to isolate the relevant host cell molecule(s) have not been successful.


The curvHDR method for gating flow cytometry samples.
High-throughput flow cytometry experiments produce hundreds of large multivariate samples of cellular characteristics. These samples require specialized processing to obtain clinically meaningful measurements. A major component of this processing is a form of cell subsetting known as gating. Manual gating is time-consuming and subjective. Good automatic and semi-automatic gating algorithms are very beneficial to high-throughput flow cytometry. We develop a statistical procedure, named curvHDR, for automatic and semi-automatic gating. The method combines the notions of significant high negative curvature regions and highest density regions and has the ability to adapt well to human-perceived gates. The underlying principles apply to dimension of arbitrary size, although we focus on dimensions up to three. Accompanying software, compatible with contemporary flow cytometry infor-matics, is developed. The method is seen to adapt well to nuances in the data and, to a reasonable extent, match human perception of useful gates. It offers big savings in human labour when processing high-throughput flow cytometry data whilst retaining a good degree of efficacy.


A critical evaluation of enzyme immunoassays for detection of antinuclear autoantibodies of defined specificities. I. Precision, sensitivity, and specificity.
To determine the performance characteristics of enzyme-based immunoassay (EIA) kits for the detection of antinuclear and other autoantibodies of defined specificities. Nine manufacturers of EIA kits to detect antibodies of defined specificities participated in a study in which they received coded sera from the Centers for Disease Control and Prevention. These coded sera contained different dilutions of antibody of one specificity mixed with sera containing antibodies of other specificities. The manufacturers were asked to use their standard technology to determine antibody content and send the data to a committee of the International Union of Immunological Societies for analysis. The data were analyzed for sensitivity and specificity in the detection of anti-double-stranded DNA (anti-dsDNA), anti-single-stranded DNA, antihistone, anti-Sm, anti-U1 RNP, anti-SSA/Ro, anti-SSB/La, anti-Scl-70 (DNA topoisomerase I), anticentromere, and anti-Jo-1 antibodies. In addition, replicate samples were included in the coded sera to evaluate the precision of each EIA method. Lack of sensitivity and specificity was most evident in the anti-dsDNA and anti-Sm kits, although 2 kits for anti-dsDNA achieved acceptable sensitivity and specificity. Generally, anti-SSA/Ro, anti-SSB/La, anti-Scl-70, anticentromere, and anti-Jo-1 kits performed well. Many false-positive results were obtained with a multiple myeloma serum containing cryoprecipitates, but multiple myeloma sera without cryoprecipitates presented no problem in the EIA system. Precision, based on evaluation of replicate samples, varied from very good to poor. No single manufacturer was clearly superior to others in terms of their products' overall sensitivity, specificity, and precision. Areas that needed improvement were in kits for the detection of antibodies to dsDNA and to Sm antigen. Some EIA kits achieved good sensitivity and specificity. Individual manufacturers were informed of the performance of their respective kits so they could take measures to correct perceived deficiencies and thus improve the reliability of a group of important diagnostic assays used in the evaluation of systemic rheumatic diseases.


Characterization of the human urinary proteome: a method for high-resolution display of urinary proteins on two-dimensional electrophoresis gels with a yield of nearly 1400 distinct protein spots.
The abundance profile of the human urinary proteome is known to change as a result of diseases or drug toxicities, particularly of those affecting the kidney and the urogenital tract. A consequence of such insults is the ability to identify proteins in urine, which may be useful as quantitative biomarkers. To succeed in discovering them, reproducible urine sample preparation methods and good protein resolution in two-dimensional electrophoresis (2-DE) gels for parallel semiquantitative protein measurements are desirable. Here, we describe a protein fractionation strategy enriching proteins of molecular masses (M(r)) lower than 30 kDa in a fraction separate from larger proteins. The fraction containing proteins with M(r)s higher than 30 kDa was subsequently subjected to immunoaffinity subtraction chromatography removing most of the highly abundant albumin and immunoglobulin G. Following 2-DE display, superior protein spot resolution was observed. Subsequent high-throughput mass spectrometry analysis of ca. 1400 distinct spots using matrix-assisted laser desorption/ionization-time of flight peptide mass fingerprinting and liquid chromatography-electrospray ionization tandem mass spectrometry lead to the successful identification of 30% of the proteins. As expected from high levels of post-translational modifications in most urinary proteins and the presence of proteolytic products, ca. 420 identified spots collapsed into 150 unique protein annotations. Only a third of the proteins identified in this study are described as classical plasma proteins in circulation, which are known to be relatively abundant in urine despite their retention to a large extent in the glomerular blood filtration process. As a proof of principle that our urinary proteome display effort holds promise for biomarker discovery, proteins isolated from the urine of a renal cell carcinoma patient were profiled prior to and after nephrectomy. Particularly, the decrease in abundance of the kininogen 2-DE gel spot train in urine after surgery was striking.


Characterisation of the DNA-polymerase-alpha-primase complex from the silk glands of Bombyx mori.
Silk gland cells of Bombyx mori undergo chromosomal endoduplication throughout larval development. The DNA content of both posterior and middle silk gland nuclei increased by 300,000 times the haploid genomic content, amounting to 18 rounds of replication. The DNA doubling time is approximately 48 h and 24 h during the fourth and fifth instars of larval development. However, DNA content does not change during the interim moult. Concomitant with DNA content, DNA polymerase activity also increases as development progressed. Enzyme activity is predominantly due to DNA polymerase alpha with no detectable level of polymerase beta. DNA polymerase alpha from silk gland extracts was purified to homogeneity (using a series of columns involving ion-exchange, gel-filtration and affinity chromatography), resulting in a 4000-fold increase in specific activity. The enzyme is a heterogeneous multimer of high molecular mass, and the catalytic (polymerase) activity is resident in the 180-kDa subunit. The enzyme shows a pI of 6.2 and the Km values for the dNTP vary over 5-16 microM. The polymerase is tightly associated with primase activity and initiates primer synthesis in the presence of ribonucleoside triphosphates on a single-stranded DNA template. The primase activity is resident in the 45-kDa subunit. The enzyme is devoid of any detectable exonuclease activity. The abundance of DNA polymerase alpha in silk glands and its strong association with the nuclear matrix suggest a role in the DNA endoduplication process.


Suppression of star formation in the galaxy NGC 253 by a starburst-driven molecular wind.
The under-abundance of very massive galaxies in the Universe is frequently attributed to the effect of galactic winds. Although ionized galactic winds are readily observable, most of the expelled mass (that is, the total mass flowing out from the nuclear region) is likely to be in atomic and molecular phases that are cooler than the ionized phases. Expanding molecular shells observed in starburst systems such as NGC 253 (ref. 12) and M 82 (refs 13, 14) may facilitate the entrainment of molecular gas in the wind. Although shell properties are well constrained, determining the amount of outflowing gas emerging from such shells and the connection between this gas and the ionized wind requires spatial resolution better than 100 parsecs coupled with sensitivity to a wide range of spatial scales, a combination hitherto not available. Here we report observations of NGC 253, a nearby starburst galaxy (distance ∼ 3.4 megaparsecs) known to possess a wind, that trace the cool molecular wind at 50-parsec resolution. At this resolution, the extraplanar molecular gas closely tracks the Hα filaments, and it appears to be connected to expanding molecular shells located in the starburst region. These observations allow us to determine that the molecular outflow rate is greater than 3 solar masses per year and probably about 9 solar masses per year. This implies a ratio of mass-outflow rate to star-formation rate of at least 1, and probably ∼3, indicating that the starburst-driven wind limits the star-formation activity and the final stellar content.


Clinical Outcomes in Patients With Type 2 Diabetes Mellitus and Peripheral Artery Disease: Results From the EXSCEL Trial.
Recent trials have identified anti-diabetes mellitus agents that lower major adverse cardiovascular event (MACE) rates, although some increase rates of lower-extremity amputation (LEA). Patients with peripheral artery disease (PAD) have greater incidence of diabetes mellitus and risk for LEA, prompting this investigation of clinical outcomes in patients with diabetes mellitus and PAD in the EXSCEL trial (Exenatide Study of Cardiovascular Event Lowering). EXSCEL evaluated the effects of once-weekly exenatide (a GLP-1 [glucagon-like peptide-1] receptor agonist) versus placebo on the rates of the primary composite MACE end point (cardiovascular death, myocardial infarction, or stroke) among patients with type 2 diabetes mellitus. In this post hoc analysis, we assessed the association of baseline PAD with rates of MACE, LEA, and the effects of exenatide versus placebo in patients with and without PAD. EXSCEL included 2800 patients with PAD (19% of the trial population). These individuals had higher unadjusted and adjusted rates of MACE compared with patients without PAD (13.6% versus 11.4%, respectively) as well as a higher adjusted hazard ratio (adjusted hazard ratio, 1.13 [95% CI, 1.00-1.27]; P=0.047). Patients with PAD had higher all-cause mortality (adjusted hazard ratio 1.38 [95% CI, 1.20-1.60]; P<0.001) and more frequent LEA (adjusted hazard ratio 5.48 [95% CI, 4.16-7.22]; P<0.001). Patients treated with exenatide or placebo had similar rates of MACE and LEA, regardless of PAD status. EXSCEL participants with PAD had higher rates of all-cause mortality and LEA compared with those without PAD. There were no differences in MACE or LEA rates with exenatide versus placebo. Clinical Trial Registration URL: https://www.clinicaltrials.gov. Unique identifier: NCT01144338.


Helping Babies Breathe (HBB) training: What happens to knowledge and skills over time?
The first minutes after birth are critical to reducing neonatal mortality. Helping Babies Breathe (HBB) is a simulation-based neonatal resuscitation program for low resource settings. We studied the impact of initial HBB training followed by refresher training on the knowledge and skills of the birth attendants in facilities. We conducted HBB trainings in 71 facilities in the NICHD Global Network research sites (Nagpur and Belgaum, India and Eldoret, Kenya), with a 6:1 ratio of facility trainees to Master Trainers (MT). Because of staff turnover, some birth attendants (BA) were trained as they joined the delivery room staff, after the initial training was completed (catch-up initial training). We compared pass rates for skills and knowledge pre- and post- initial HBB training and following refresher training among active BAs. An Objective Structured Clinical Examination (OSCE) B tested resuscitation skill retention by comparing post-initial training performance with pre-refresher training performance. We identified factors associated with loss of skills in pre-refresher training performance using multivariable logistic regression analysis. Daily bag and mask ventilation practice, equipment checks and supportive supervision were stressed as part of training. One hundred five MT (1.6 MT per facility) conducted initial and refresher HBB trainings for 835 BAs; 76% had no prior resuscitation training. Initial training improved knowledge and skills: the pass percentage for knowledge tests improved from 74 to 99% (p < 0.001). Only 5% could ventilate a newborn mannequin correctly before initial training but 97% passed the post-initial ventilation training test (p < 0.0001) and 99% passed the OSCE B resuscitation evaluation. During pre-refresher training evaluation, a mean of 6.7 (SD 2.49) months after the initial training, 99% passed the knowledge test, but the successful completion rate fell to 81% for the OSCE B resuscitation skills test. Characteristics associated with deterioration of resuscitation skills were BAs from tertiary care facilities, no prior resuscitation training, and the timing of training (initial vs. catch-up training). HBB training significantly improved neonatal resuscitation knowledge and skills. However, skills declined more than knowledge over time. Ongoing skills practice and monitoring, more frequent retesting, and refresher trainings are needed to maintain neonatal resuscitation skills. ClinicalTrials.gov Identifier: NCT01681017 ; 04 September 2012, retrospectively registered.


Multi-targeted priming for genome-wide gene expression assays.
Complementary approaches to assaying global gene expression are needed to assess gene expression in regions that are poorly assayed by current methodologies. A key component of nearly all gene expression assays is the reverse transcription of transcribed sequences that has traditionally been performed by priming the poly-A tails on many of the transcribed genes in eukaryotes with oligo-dT, or by priming RNA indiscriminately with random hexamers. We designed an algorithm to find common sequence motifs that were present within most protein-coding genes of Saccharomyces cerevisiae and of Neurospora crassa, but that were not present within their ribosomal RNA or transfer RNA genes. We then experimentally tested whether degenerately priming these motifs with multi-targeted primers improved the accuracy and completeness of transcriptomic assays. We discovered two multi-targeted primers that would prime a preponderance of genes in the genomes of Saccharomyces cerevisiae and Neurospora crassa while avoiding priming ribosomal RNA or transfer RNA. Examining the response of Saccharomyces cerevisiae to nitrogen deficiency and profiling Neurospora crassa early sexual development, we demonstrated that using multi-targeted primers in reverse transcription led to superior performance of microarray profiling and next-generation RNA tag sequencing. Priming with multi-targeted primers in addition to oligo-dT resulted in higher sensitivity, a larger number of well-measured genes and greater power to detect differences in gene expression. Our results provide the most complete and detailed expression profiles of the yeast nitrogen starvation response and N. crassa early sexual development to date. Furthermore, our multi-targeting priming methodology for genome-wide gene expression assays provides selective targeting of multiple sequences and counter-selection against undesirable sequences, facilitating a more complete and precise assay of the transcribed sequences within the genome.


Small fields measurements with radiochromic films.
The small fields in radiotherapy are widely used due to the development of techniques such as intensity-modulated radiotherapy and stereotactic radio surgery. The measurement of the dose distributions for small fields is a challenge. A perfect dosimeter should be independent of the radiation energy and the dose rate and should have a negligible volume effect. The radiochromic (RC) film characteristics fit well to these requirements. However, the response of RC films and their digitizing processes present a significant spatial inhomogeneity problem. The present work uses a method for two-dimensional (2D) measurement with RC films based on the reduction of the spatial inhomogeneity of both the film and the film digitizing process. By means of registering and averaging several measurements of the same field, the inhomogeneities are mostly canceled. Measurements of output factors (OFs), dose profiles (in-plane and cross-plane), and 2D dose distributions are presented. The field sizes investigated are 0.5 × 0.5 cm(2), 0.7 × 0.7 cm(2), 1 × 1 cm(2), 2 × 2 cm(2), 3 × 3 cm(2), 6 × 6 cm(2), and 10 × 10 cm(2) for 6 and 15 MV photon beams. The OFs measured with the RC film are compared with the measurements carried out with a PinPoint ionization chamber (IC) and a Semiflex IC, while the measured transversal dose profiles were compared with Monte Carlo simulations. The results obtained for the OFs measurements show a good agreement with the values obtained from RC films and the PinPoint and Semiflex chambers when the field size is greater or equal than 2 × 2 cm(2). These agreements give confidence on the accuracy of the method as well as on the results obtained for smaller fields. Also, good agreement was found between the measured profiles and the Monte Carlo calculated profiles for the field size of 1 × 1 cm(2). We expect, therefore, that the presented method can be used to perform accurate measurements of small fields.


Magnetic polymer models for epigenetics-driven chromosome folding.
Epigenetics is a driving force of important and ubiquitous phenomena in nature such as cell differentiation or even metamorphosis. Opposite to its widespread role, understanding the biophysical principles that allow epigenetics to control and rewire gene regulatory networks remains an open challenge. In this work we study the effects of epigenetic modifications on the spatial folding of chromosomes-and hence on the expression of the underlying genes-by mapping the problem to a class of models known as magnetic polymers. In this work we show that a first order phase transition underlies the simultaneous spreading of certain epigenetic marks and the conformational collapse of a chromosome. Further, we describe Brownian dynamics simulations of the model in which the topology of the polymer and thermal fluctuations are fully taken into account and that confirm our mean field predictions. Extending our models to allow for nonequilibrium terms yields new stable phases which qualitatively agrees with observations in vivo. Our results show that statistical mechanics techniques applied to models of magnetic polymers can be successfully exploited to rationalize the outcomes of experiments designed to probe the interplay between a dynamic epigenetic landscape and chromatin organization.


The C-terminal end of the Trypanosoma brucei editing deaminase plays a critical role in tRNA binding.
Adenosine to inosine editing at the wobble position allows decoding of multiple codons by a single tRNA. This reaction is catalyzed by adenosine deaminases acting on tRNA (ADATs) and is essential for viability. In bacteria, the anticodon-specific enzyme is a homodimer that recognizes a single tRNA substrate (tRNA(Arg)(ACG)) and can efficiently deaminate short anticodon stem-loop mimics of this tRNA in vitro. The eukaryal enzyme is composed of two nonidentical subunits, ADAT2 and ADAT3, which upon heterodimerization, recognize seven to eight different tRNAs as substrates, depending on the organism, and require a full-length tRNA for activity. Although crystallographic data have provided clues to why the bacterial deaminase can utilize short substrates, residues that provide substrate binding and recognition with the eukaryotic enzymes are not currently known. In the present study, we have used a combination of mutagenesis, binding studies, and kinetic analysis to explore the contribution of individual residues in Trypanosoma brucei ADAT2 (TbADAT2) to tRNA recognition. We show that deletion of the last 10 amino acids at the C terminus of TbADAT2 abolishes tRNA binding. In addition, single alanine replacements of a string of positively charged amino acids (KRKRK) lead to binding defects that correlate with losses in enzyme activity. This region, which we have termed the KR-domain, provides a first glance at key residues involved in tRNA binding by eukaryotic tRNA editing deaminases.


The politics of decarbonization and the catalytic impact of subnational climate experiments.
The Paris Agreement of 2015 marks a formal shift in global climate change governance from an international legal regime that distributes state commitments to solve a collective action problem to a catalytic mechanism to promote and facilitate transformative pathways to decarbonization. It does so through a system of nationally determined contributions, monitoring and ratcheting up of commitments, and recognition that the practice of climate governance already involved an array of actors and institutions at multiple scales. In this article, we develop a framework that focuses on the politics of decarbonization to explore policy pathways and mechanisms that can disrupt carbon lock-in through these diverse, decentralized responses. It identifies political mechanisms-normalization, capacity building, and coalition building-that contribute to the scaling and entrenchment of discrete decarbonization initiatives within or across jurisdictions, markets, and practices. The role for subnational (municipal, state/provincial) climate governance experiments in this new context is especially profound. Drawing on such cases, we illustrate the framework, demonstrate its utility, and show how its political analysis can provide insight into the relationship between climate governance experiments and the formal global response as well as the broader challenge of decarbonization.


Global conformational dynamics of a Y-family DNA polymerase during catalysis.
Replicative DNA polymerases are stalled by damaged DNA while the newly discovered Y-family DNA polymerases are recruited to rescue these stalled replication forks, thereby enhancing cell survival. The Y-family DNA polymerases, characterized by low fidelity and processivity, are able to bypass different classes of DNA lesions. A variety of kinetic and structural studies have established a minimal reaction pathway common to all DNA polymerases, although the conformational intermediates are not well defined. Furthermore, the identification of the rate-limiting step of nucleotide incorporation catalyzed by any DNA polymerase has been a matter of long debate. By monitoring time-dependent fluorescence resonance energy transfer (FRET) signal changes at multiple sites in each domain and DNA during catalysis, we present here a real-time picture of the global conformational transitions of a model Y-family enzyme: DNA polymerase IV (Dpo4) from Sulfolobus solfataricus. Our results provide evidence for a hypothetical DNA translocation event followed by a rapid protein conformational change prior to catalysis and a subsequent slow, post-chemistry protein conformational change. Surprisingly, the DNA translocation step was induced by the binding of a correct nucleotide. Moreover, we have determined the directions, rates, and activation energy barriers of the protein conformational transitions, which indicated that the four domains of Dpo4 moved in a synchronized manner. These results showed conclusively that a pre-chemistry conformational change associated with domain movements was too fast to be the rate-limiting step. Rather, the rearrangement of active site residues limited the rate of correct nucleotide incorporation. Collectively, the conformational dynamics of Dpo4 offer insights into how the inter-domain movements are related to enzymatic function and their concerted interactions with other proteins at the replication fork.


Task-independent and task-specific age effects on brain activity during working memory, visual attention and episodic retrieval.
It is controversial whether the effects of aging on various cognitive functions have the same common cause or several different causes. To investigate this issue, we scanned younger and older adults with functional magnetic resonance imaging (fMRI) while performing three different tasks: working memory, visual attention and episodic retrieval. There were three main results. First, in all three tasks, older adults showed weaker occipital activity and stronger prefrontal and parietal activity than younger adults. The occipital reduction is consistent with the view that sensory processing decline is a common cause in cognitive aging, and the prefrontal increase may reflect functional compensation. Secondly, older adults showed more bilateral patterns of prefrontal activity than younger adults during working memory and visual attention tasks. These findings are consistent with the Hemispheric Asymmetry Reduction in Older Adults (HAROLD) model. Finally, compared to younger adults, older adults showed weaker hippocampal formation activity in all three tasks but stronger parahippocampal activity in the episodic retrieval task. The former finding suggests that age-related hippocampal deficits may have a global effect in cognition, and the latter is consistent with an age-related increase in familiarity-based recognition. Taken together, the results indicate that both common and specific factors play an important role in cognitive aging.


An image-guided precision proton radiation platform for preclinical in vivo research.
There are many unknowns in the radiobiology of proton beams and other particle beams. We describe the development and testing of an image-guided low-energy proton system optimized for radiobiological research applications. A 50 MeV proton beam from an existing cyclotron was modified to produce collimated beams (as small as 2 mm in diameter). Ionization chamber and radiochromic film measurements were performed and benchmarked with Monte Carlo simulations (TOPAS). The proton beam was aligned with a commercially-available CT image-guided x-ray irradiator device (SARRP, Xstrahl Inc.). To examine the alternative possibility of adapting a clinical proton therapy system, we performed Monte Carlo simulations of a range-shifted 100 MeV clinical beam. The proton beam exhibits a pristine Bragg Peak at a depth of 21 mm in water with a dose rate of 8.4 Gy min-1 (3 mm depth). The energy of the incident beam can be modulated to lower energies while preserving the Bragg peak. The LET was: 2.0 keV µm-1 (water surface), 16 keV µm-1 (Bragg peak), 27 keV µm-1 (10% peak dose). Alignment of the proton beam with the SARRP system isocenter was measured at 0.24 mm agreement. The width of the beam changes very little with depth. Monte Carlo-based calculations of dose using the CT image data set as input demonstrate in vivo use. Monte Carlo simulations of the modulated 100 MeV clinical proton beam show a significantly reduced Bragg peak. We demonstrate the feasibility of a proton beam integrated with a commercial x-ray image-guidance system for preclinical in vivo studies. To our knowledge this is the first description of an experimental image-guided proton beam for preclinical radiobiology research. It will enable in vivo investigations of radiobiological effects in proton beams.


Catalytically propelled 3D printed colloidal microswimmers.
Synthetic microswimmers are widely employed model systems in the studies of out-of-equilibrium phenomena. Unlike biological microswimmers which naturally occur in various shapes and forms, synthetic microswimmers have so far been limited almost exclusively to spherical shapes. Here, we exploit 3D printing to produce microswimmers with complex shapes in the colloidal size regime. We establish the flexibility of 3D printing by two-photon polymerisation to produce particles smaller than 10 microns with a high-degree of shape complexity. We further demonstrate that 3D printing allows control over the location of the active site through orienting the particles in different directions during printing. We verify that particles behave colloidally by imaging their motion in the passive and active states and by investigating their mean square displacement. In addition, we find that particles exhibit shape-dependant behavior, thereby demonstrating the potential of our method to launch a wide-range of in-depth studies into shape-dependent active motion and behaviour.


Modeling of temperature and excitation dependences of efficiency in an InGaN light-emitting diode.
The changes in excitation dependence of efficiency with temperature are modeled for a wurtzite InGaN light-emitting diode. The model incorporates bandstructure changes with carrier density because of screening of quantum-confined Stark effect. Bandstructure is computed by solving Poisson and k · p equations in the envelope approximation. The information is used in a dynamical model for populations in momentum-resolved electron and hole states. Application of the approach shows the interplay of quantum-well and barrier emissions giving rise to shape changes in efficiency versus current density with changing temperature, as observed in some experiments.


Possible axonal regrowth in late recovery from the minimally conscious state.
We used diffusion tensor imaging (DTI) to study 2 patients with traumatic brain injury. The first patient recovered reliable expressive language after 19 years in a minimally conscious state (MCS); the second had remained in MCS for 6 years. Comparison of white matter integrity in the patients and 20 normal subjects using histograms of apparent diffusion constants and diffusion anisotropy identified widespread altered diffusivity and decreased anisotropy in the damaged white matter. These findings remained unchanged over an 18-month interval between 2 studies in the first patient. In addition, in this patient, we identified large, bilateral regions of posterior white matter with significantly increased anisotropy that reduced over 18 months. In contrast, notable increases in anisotropy within the midline cerebellar white matter in the second study correlated with marked clinical improvements in motor functions. This finding was further correlated with an increase in resting metabolism measured by PET in this subregion. Aberrant white matter structures were evident in the second patient's DTI images but were not clinically correlated. We propose that axonal regrowth may underlie these findings and provide a biological mechanism for late recovery. Our results are discussed in the context of recent experimental studies that support this inference.


Are revisions of patellofemoral arthroplasties more like primary or revision TKAs.
Patellofemoral arthroplasty (PFA) can be considered in patients with patellofemoral disease. However, the use of partial arthroplasty often causes concern among clinicians and patients that revision to total knee arthroplasty (TKA) will be needed and, if so, whether this revision will be straightforward or more complicated. We set out to determine if conversion of a PFA to a TKA was more similar to a primary or to a revision TKA in terms of surgical characteristics, knee scores, range of motion, and complications. Between 2001 and 2008, we revised 21 PFAs to TKAs, all of which were available for followup at a minimum of 5 years (median, 6 years; range, 5-12 years). These patients were matched one-to-one by age, sex, body mass index, length of followup, and preoperative Knee Society Scores (KSS) to 21 primary and 21 revision TKAs. We analyzed operative time and amount of blood loss. Clinical outcomes assessed were range of motion and KSS. Blood loss (405 mL versus 460 mL versus 900 mL; odds/hazard ratio, 1.33, 95% confidence interval [CI], 0.3-5.85; p=0.14 for primary TKA versus revision PFA and odds/hazard ratio, 0.13, 95% CI, 0.03-0.52; p<0.01 for revision PFA versus revision TKA) and operative time (52 minutes versus 72 minutes versus 115 minutes; odds/hazard ratio, 5.45, 95% CI, 1.23-27.4; p=0.02 for primary TKA versus revision PFA and odds/hazard ratio, 0.5, 95% CI, 0.01-0.44; p<0.001 for revision PFA versus revision TKA) were not different between the primary TKA and revision PFA groups but higher in the revision TKA group. KSS (knee and function) were higher in the primary TKA group (92 [range, 60-100] and 91 [range, 65-100]) than they were in the revision PFA (85 [range, 40-100] and 85 [range, 30-100]) and revision TKA groups (75 [range, 30-100] and 68 [range, 25-100]; p<0.001). Flexion was better in the primary TKA (125 [range, 105-130]) and revised PFA (120 [range, 100-130]) groups than the revision TKA group (105 [range, 80-115]; p=0.0013). There were more complications in the revision PFA group (two of 21) compared with the primary TKA group (zero of 21, p=0.005) but not compared with the revision TKA group (three of 21; p=0.85). With the numbers available, we found that revising a PFA is comparable to a primary TKA in regard to surgical characteristics and postoperative clinical outcomes (including knee scores and range of motion), and both are superior to revision TKA, although the frequency of complications was higher in the revision PFA group than it was in the primary TKA group. The majority of patients undergoing revision of a PFA to a TKA can be treated with a standard implant. Level III, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


A holographic interferometer for measuring radiation energy deposition profiles in transparent liquids.
An apparatus has been designed for real-time and double-exposure holographic interferometry to determine radiation absorbed dose distributions in transparent liquids. The change in refractive index of the liquid due to a temperature rise after irradiation is measured interferometrically. In a cylindrically symmetrical radiation field, the dose distribution can be computed from data supplied by the reconstruction of the holographic interferogram taken as side-view profile of the change in optical pathlength. Relatively inexpensive components such as a low-powered He-Ne laser together with a conventional photographic shutter and low-cost mirrors and lenses were used. The mathematical procedure for unfolding the three-dimensional dose distribtion is described, and an example is given for use with a high-intensity, pulsed, 2-MV electron source.


Bacterial community composition over a dry winter in meso- and eutrophic Portuguese water bodies.
In order to investigate the bacterial diversity in a number of rivers, reservoirs and lakes in northern and central Portugal during the winter of 2004/5 (atypically dry), we applied molecular methodologies, namely denaturing gradient gel electrophoresis with primers targeting fractions of the bacterial 16S rRNA gene. Environmental parameters such as pH, conductivity, inorganic nutrients, total suspended solids and chlorophyll a were determined in order to characterize the trophic status of the studied water bodies. We found water bodies with oligotrophic to hypereutrophic characteristics. Organisms belonging to the Bacteroidetes and Alphaproteobacteria were found at the highest pH environment. Bacteroidetes were also related to high nutrient concentrations. Verrucomicrobia were associated with the most oligotrophic reservoir and low pH values. Actinobacteria were present in all samples from lakes and reservoirs, indicating its preference for lentic water bodies. Cyanobacteria dominance was related to high pH and conductivity levels. In general the conductivity values recorded in winter 2005 were the highest over recent years and chlorophyll a also reached very high levels. The data emphasize an enhanced risk of eutrophication for the studied water bodies, especially in the subsequent months when the temperature rises.


Hepatitis C virus vaccines in the era of new direct-acting antivirals.
Hepatitis C virus (HCV) infection is a major global health problem as it has a high propensity for establishing chronicity. Chronic HCV carriers are at risk of developing severe liver disease including fibrosis, cirrhosis and liver cancer. While treatment has considerably improved over the years, therapy is still only partially effective, and is plagued by side effects, which contribute to treatment failure and is expensive to manage. The drug development pipeline contains several compounds that hold promise to achieve the goal of a short and more tolerable therapy, and are also likely to improve treatment response rates. It remains to be seen, however, how potent antiviral drug cocktails will affect the hepatitis C burden worldwide. In resource-poor environments, considerable costs, inadequate infrastructure for medical supervision and distribution may diminish the impact of future therapies. Consequently, development of novel therapeutic and prophylactic strategies is imperative to contain HCV infection globally.


Sequence variant on 8q24 confers susceptibility to urinary bladder cancer.
We conducted a genome-wide SNP association study on 1,803 urinary bladder cancer (UBC) cases and 34,336 controls from Iceland and The Netherlands and follow up studies in seven additional case-control groups (2,165 cases and 3,800 controls). The strongest association was observed with allele T of rs9642880 on chromosome 8q24, 30 kb upstream of MYC (allele-specific odds ratio (OR) = 1.22; P = 9.34 x 10(-12)). Approximately 20% of individuals of European ancestry are homozygous for rs9642880[T], and their estimated risk of developing UBC is 1.49 times that of noncarriers. No association was observed between UBC and the four 8q24 variants previously associated with prostate, colorectal and breast cancers, nor did rs9642880 associate with any of these three cancers. A weaker signal, but nonetheless of genome-wide significance, was captured by rs710521[A] located near TP63 on chromosome 3q28 (allele-specific OR = 1.19; P = 1. 15 x 10(-7)).


Polygenic inheritance of adolescent idiopathic scoliosis: a study of extended families in Utah.
A heritability study of 69 extended Utah families with a history of adolescent idiopathic scoliosis (AIS) indicates that AIS is a polygenic, multifactorial condition. Each family reported a history of AIS within four generations; a total of 247 individuals were confirmed via X-rays and medical records to have AIS. Coefficient of kinship was more than 25 standard deviations higher for these 69 families than for the general population. Excluding all probands and assuming autosomal dominant inheritance, 1,260 individuals over the age of 16 were determined to be at risk for AIS because they have a parent with AIS. Assuming 50% of these individuals carry the allele, estimated penetrance in at-risk males is approximately 9%, and estimated penetrance in at-risk females is approximately 29%. Recurrence risk in relatives decreases as the degree of relationship to the affected individual becomes more distant; however, the lowest recurrence risk calculated, for third-degree relatives, is still an average of 9%, well above the general population's risk. Onset of AIS appears to be inherited separate from curve pattern and severity. In a study of phenotypes in 36 of the families, the affected individuals were consistent in either curve severity or curve pattern, but not both. It is unclear whether severity or pattern is more heritable, but it is possible that the location of the curve on the spine is the most heritable trait of the phenotype. The study demonstrates the genetic complexity of AIS, including the low penetrance of its cumulative alleles and variable expression.


Carcinoembryonic antigen-related cell adhesion molecule 2 controls energy balance and peripheral insulin action in mice.
The carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a transmembrane glycoprotein with pleotropic functions, including clearance of hepatic insulin. We investigated the functions of the related protein CEACAM2, which has tissue-specific distribution (kidney, uterus, and crypt epithelia of intestinal tissues), in genetically modified mice. Ceacam2-null mice (Cc2-/-) were generated from a 129/SvxC57BL/6J background. Female mice were assessed by hyperinsulinemic-euglycemic clamp analysis and indirect calorimetry and body fat composition was measured. Cc2-/- mice and controls were fed as pairs, given insulin tolerance tests, and phenotypically characterized. Female, but not male Cc2-/- mice exhibited obesity that resulted from hyperphagia and reduced energy expenditure. Pair feeding experiments showed that hyperphagia led to peripheral insulin resistance. Insulin action was normal in liver but compromised in skeletal muscle of female Cc2-/- mice; the mice had incomplete fatty acid oxidation and impaired glucose uptake and disposal. The mechanism of hyperphagia in Cc2-/- mice is not clear, but appears to result partly from increased hyperinsulinemia-induced hypothalamic fatty acid synthase levels and activity. Hyperinsulinemia was caused by increased insulin secretion. In mice, CEACAM2 is expressed by the hypothalamus. Cc2-/- mice develop obesity from hyperphagia and reduced energy expenditure, indicating its role in regulating energy balance and insulin sensitivity.


Age-related changes in blood pressure.
This report is based on three different representative population samples of a total of 1304 men (50-79 years old) and 1246 women (38-79 years old) observed for up to 12 years. Subjects' consumption of antihypertensive drugs and blood pressure levels in subjects with and without such treatment are presented. The prevalence of treatment with antihypertensive drugs (including beta-blockers and diuretics for other indications) increased from 2% at age 50 years to 37% at 79 years of age among the men and from 1% at 38 years to 61% at 79 years of age among the women. The mean systolic/diastolic blood pressure in untreated subjects increased from 138/91 mm Hg at age 50 years to 159/91 mm Hg at age 70 years in the men and from 123/79 mm Hg at age 38 years to 168/93 mm Hg at age 70 years in the women. At age 79 years the mean systolic/diastolic blood pressure was 155/83 mm Hg in the men and 161/85 mm Hg in the women. In a longitudinal follow-up of reexamined subjects, there was an increase in systolic blood pressure levels up to age 75 years and a reduction in diastolic blood pressure after age 75 years in both sexes.


Electrospray droplet exposure to organic vapors: metal ion removal from proteins and protein complexes.
The exposure of aqueous nanoelectrospray droplets to various organic vapors can dramatically reduce sodium adduction on protein ions in positive ion mass spectra. Volatile alcohols, such as methanol, ethanol, and isopropanol lead to a significant reduction in sodium ion adduction but are not as effective as acetonitrile, acetone, and ethyl acetate. Organic vapor exposure in the negative ion mode, on the other hand, has essentially no effect on alkali ion adduction. Evidence is presented to suggest that the mechanism by which organic vapor exposure reduces alkali ion adduction in the positive mode involves the depletion of alkali metal ions via ion evaporation of metal ions solvated with organic molecules. The early generation of metal/organic cluster ions during the droplet desolvation process results in fewer metal ions available to condense on the protein ions formed via the charged residue mechanism. These effects are demonstrated with holomyoglobin ions to illustrate that the metal ion reduction takes place without detectable protein denaturation, which might be revealed by heme loss or an increase in charge state distribution. No evidence is observed for denaturation with exposure to any of the organic vapors evaluated in this work.


Molecular epidemiological survey of rotaviruses in sewage by reverse transcriptase seminested PCR and restriction fragment length polymorphism assay.
Rotavirus double-stranded RNA was detected directly in sewage treatment plant samples over a 1-year period by reverse transcription followed by PCR amplification of the VP7 gene and Southern blot hybridization. The presence of naturally occurring rotaviruses was demonstrated in 42% of raw sewage samples and in 67% of treated effluent samples. Amplified viral sequences were analyzed by restriction enzymes. Ten different restriction profiles were characterized, most of which were found in treated effluent samples. A mixture of restriction profiles was observed in 75% of contaminated effluent samples. The profiles were compared with those obtained from human rotavirus isolates involved in infections in children from the same area (six different profiles were detected). Five identical viral sequences were detected in both environmental and clinical samples. Restriction profiles were also compared to profiles from known genomic sequences of human and animal viruses. Both human and animal origins of rotavirus contamination of water seemed likely.


Functional imaging of changes in cerebellar activity related to learning during a novel eye-hand tracking task.
Coordination between the eyes and the hand is likely to be based on a process of motor learning, so that the interactions between the two systems can be accurately controlled. By using an unusual tracking task we measured the change in brain activation levels, as recorded with 3T functional magnetic resonance imaging (fMRI), between naïve human subjects and the same subjects after a period of extended training. Initially the performance of the two groups was similar. One subject group was then trained in a synchronous, coordinated, eye-hand task; the other group trained with a 304 ms temporal offset between hand and eye tracking movements. After training, different patterns of performance were observed for the groups, and different functional activation profiles. Significant change in the relationship between functional activation levels and eye-hand task conditions was predominantly restricted to visuo-motor areas of the lateral and vermal cerebellum. In an additional test with one of the subject groups, we show that there was increased cerebellar activation after learning, irrespective of change in performance error. These results suggest that two factors contribute to the measured blood oxygen level-dependent (BOLD) signal. One declined with training and may be directly related to performance error. The other increased after training, in the test conditions nearest to the training condition, and may therefore be related to acquisition of experience in the task. The loci of activity changes suggest that improved performance is because of selective modified processing of ocular and manual control signals within the cerebellum. These results support the suggestion that coordination between eye and hand movement is based on an internal model acquired by the cerebellum that provides predictive signals linking the control of the two effectors.


Feedback control of AHR signalling regulates intestinal immunity.
The aryl hydrocarbon receptor (AHR) recognizes xenobiotics as well as natural compounds such as tryptophan metabolites, dietary components and microbiota-derived factors, and it is important for maintenance of homeostasis at mucosal surfaces. AHR activation induces cytochrome P4501 (CYP1) enzymes, which oxygenate AHR ligands, leading to their metabolic clearance and detoxification. Thus, CYP1 enzymes have an important feedback role that curtails the duration of AHR signalling, but it remains unclear whether they also regulate AHR ligand availability in vivo. Here we show that dysregulated expression of Cyp1a1 in mice depletes the reservoir of natural AHR ligands, generating a quasi AHR-deficient state. Constitutive expression of Cyp1a1 throughout the body or restricted specifically to intestinal epithelial cells resulted in loss of AHR-dependent type 3 innate lymphoid cells and T helper 17 cells and increased susceptibility to enteric infection. The deleterious effects of excessive AHR ligand degradation on intestinal immune functions could be counter-balanced by increasing the intake of AHR ligands in the diet. Thus, our data indicate that intestinal epithelial cells serve as gatekeepers for the supply of AHR ligands to the host and emphasize the importance of feedback control in modulating AHR pathway activation.


Systemic TNF blockade does not modulate synovial expression of the pro-inflammatory mediator HMGB1 in rheumatoid arthritis patients--a prospective clinical study.
High-mobility group box chromosomal protein 1 (HMGB1) has recently been identified as an endogenous mediator of arthritis. TNF and IL-1beta, pivotal cytokines in arthritis pathogenesis, both have the ability to induce the release of HMGB1 from myeloid and dendritic cells. It was, therefore, decided to investigate whether treatment based on TNF blockade in rheumatoid arthritis (RA) affects the expression of synovial HMGB1. Repeated arthroscopy-guided sampling of synovial tissue was performed in nine patients with RA before and nine weeks after initiation of anti-TNF mAb (infliximab) therapy. Synovial biopsy specimens were analysed for HMGB1 protein by immunohistochemical staining and for HMGB1 mRNA expression by real-time reverse transcriptase PCR (RT-PCR). Statistical evaluations were based on Wilcoxon's signed rank tests or Spearman rank sum tests. Aberrant, extranuclear HMGB1 and constitutive nuclear HMGB1 expression, with histological signs of inflammation, were evident in all biopsies obtained before infliximab therapy. Signs of inflammation were still evident in the second biopsies obtained nine weeks after initiation of infliximab therapy. The cytoplasmic and extracellular expression of HMGB1 decreased in five patients, remained unchanged in one patient and increased in three patients, making the overall change in HMGB1 protein expression not significant. No correlation between the clinical response, as measured by disease activity score calculated for 28 joints (DAS28) or the American College of Rheumatology response criteria (ACR 20, 50, and 70), and the direction of change of HMGB1 expression in individual patients could be discerned. In addition, infliximab therapy did not alter HMGB1 mRNA synthesis. Pro-inflammatory HMGB1 expression during rheumatoid synovitis was not consistently influenced by TNF-blocking therapy with infliximab. This suggests that TNF is not the main inducer of extranuclear HMGB1 during synovitis and that HMGB1 may represent a TNF-independent molecule that could be considered as a possible target for future therapeutic intervention in RA.


Occurrence of Potato Cyst Nematode, Globodera rostochiensis, on Potato in the Saint-Amable Region, Quebec, Canada.
Potato (Solanum tuberosum L. subsp. tuberosum) is one of the most important food crops in Canada. Potato cyst nematodes, Globodera rostochiensis (Wollenweber, 1923) Skarbilovich, 1959 and Globodera pallida (Stone, 1973) Behrens, 1975, are considered the most economically important nematode pests of potatoes worldwide and are the subject of strict quarantine regulations in many countries including Canada. In Canada, G. rostochiensis was found in the Saanich Peninsula of Vancouver Island, British Columbia and both G. rostochiensis and G. pallida are present on the island of Newfoundland (3). During August of 2006, soil and roots of potato plants collected from a 19.2-ha field in the Saint-Amable region, Quebec, were submitted to the Nematology Laboratory, Canadian Food Inspection Agency, Ottawa. Golden, spherical-shaped cysts were found associated with the roots and were also extracted from the soil. The nematodes recovered were identified by morphological and morphometric analysis and DNA analysis. Measurements and morphological observation of 60 second-stage juveniles were as follows: body length = 427.4 ± 22.0 (395 to 495) μm; stylet length = 21.3 ± 1.2 (18 to 23) μm; distance from stylet base to dorsal esophageal gland opening = 5.5 ± 0.9 (3.5 to 7.0) μm; tail length = 46.7 ± 3.7 (36 to 58) μm; hyaline tail terminus = 21.1 ± 3.1 (13 to 27) μm; and the shape of stylet basal knobs varied from rounded to lateral flattened. Observations for 35 mature cysts were: number of cuticular ridges between anus and vulval fenestra = 21 ± 6 (12 to 31); fenestral length = 18.4 ± 3.6 (11 to 30) μm; distance from anus to the edge of fenestra = 81.1 ± 30.4 (29 to 165) μm; Granek's ratio = 4.4 ± 1.6 (1.8 to 6.0), with parallel wavy cuticular ridges between anus and vulval fenestra. These observations conformed to the published description of G. rostochiensis (2). Additionally, the identification was supported by the PCR-restriction fragment length polymorphism pattern of the ITS-1 region amplified using primers 18S (5'-TTGATTACGTCCCTGCCCTTT-3') and rDNA1.58S (5'-ACGAGCCGAGTGATCCACCG-3') and digested with BstU I and a PCR product specific for G. rostochiensis (1). Sequence of a 1,190-bp PCR fragment of ribosomal DNA amplified using primers rDNA1 (5'-TTGATTACGTCCCTGCCCTTT-3') and rDNA2 (5'- TTTCACTCGCCGTTACTAAGG-3') showed 99.0 to 100% identity with published DNA sequences of the same genomic region for G. rostochiensis (4). The sequence is available from the authors upon request. The origin of the introduction of G. rostochiensis into Saint-Amable is unknown. An intensive soil survey is underway to define the infested area, and strict quarantine measures have been taken to prevent further spread of G. rostochiensis. To our knowledge, this is the first evidence of the occurrence of G. rostochiensis in Quebec, Canada. References: (1) A. Fullaondo et al. Nematology 1:157, 1999. (2) A. M. Golden et al. Proc. Helminthol. Soc. Wash. 39:64, 1972. (3) A. R. Stone et al. Plant Dis. Rep. 61:590, 1977. (4) S. A. Subbotin et al. Nematology 2:591, 2000.


Temporal modulation of spatial tactile extinction in right-brain-damaged patients.
Unilateral and bilateral electrotactile stimuli were delivered to both hands of 11 right-brain-damaged (RBD) patients with left tactile extinction and 20 healthy subjects. Bimanual stimuli could be presented simultaneously or with varying stimulus onset asynchronies (SOAs). Subjects indicated their detection of unilateral or bilateral stimuli, their judgements of whether stimuli were simultaneous or successive, and, in the latter case, which side came first. In RBD patients, extinction was maximal with simultaneous presentations and decreased as SOA increased. With short SOAs, omissions of left-sided stimuli occurred with both right-side and left-side stimulus precedence, suggesting a forward and backward interference of the right stimulus on the processing of the left stimulus within a time window of at least 100 msec. In contrast, there was no interference of the left stimulus on the detection of the right stimulus. Unlike controls, extinction patients rarely expressed simultaneity judgements, but those that were produced tended to be veridical or nearly so, like in normal controls. Whereas controls expressed generally accurate judgements of right or left precedence, patients showed a bias toward a right precedence and a maximal uncertainty between left-first and right-first choices when the left stimulus had a lead between 100 and 200 msec. The results are consistent with the hypothesis that, as in other sensory modalities, tactile extinction is associated with an abnormal persistent bias of attention toward the ipsilesional side that delays the processing of contralesional stimuli. However, the finding that both extinction and explicit judgements of simultaneity tended to occur with simultaneous bilateral stimuli suggest the presence of some residual neural capacity to detect precise temporal coincidence.


Pharmacogenomic study in patients with multiple sclerosis: Responders and nonresponders to IFN-β.
We aimed to investigate the association between polymorphisms located in type I interferon (IFN)-induced genes, genes belonging to the toll-like receptor (TLR) pathway, and genes encoding neurotransmitter receptors and the response to IFN-β treatment in patients with multiple sclerosis (MS). In a first or screening phase of the study, 384 polymorphisms were genotyped in 830 patients with MS classified into IFN-β responders (n = 416) and nonresponders (n = 414) according to clinical criteria. In a second or validation phase, the most significant polymorphisms associated with IFN-β response were genotyped in an independent validation cohort of 555 patients with MS (281 IFN-β responders and 274 nonresponders). Seven single nucleotide polymorphisms (SNPs) were selected from the screening phase for further validation: rs832032 (GABRR3; p = 0.0006), rs6597 (STUB1; p = 0.019), rs3747517 (IFIH1; p = 0.010), rs2277302 (PELI3; p = 0.017), rs10958713 (IKBKB; p = 0.003), rs2834202 (IFNAR1; p = 0.030), and rs4422395 (CXCL1; p = 0.017). None of these SNPs were significantly associated with IFN-β response when genotyped in an independent cohort of patients. Combined analysis of these SNPs in all patients with MS (N = 1,385) revealed 2 polymorphisms associated with IFN-β response: rs2277302 (PELI3; p = 0.008) and rs832032 (GABRR3; p = 0.006). These findings do not support an association between polymorphisms located in genes related to the type I IFN or TLR pathways or genes encoding neurotransmitter receptors and the clinical response to IFN-β. Nevertheless, additional genetic and functional studies of PELI3 and GABRR3 are warranted.


Analysis of the cytochrome c oxidase subunit II (COX2) gene in giant panda, Ailuropoda melanoleuca.
The giant panda, Ailuropoda melanoleuca (Ursidae), has a unique bamboo-based diet; however, this low-energy intake has been sufficient to maintain the metabolic processes of this species since the fourth ice age. As mitochondria are the main sites for energy metabolism in animals, the protein-coding genes involved in mitochondrial respiratory chains, particularly cytochrome c oxidase subunit II (COX2), which is the rate-limiting enzyme in electron transfer, could play an important role in giant panda metabolism. Therefore, the present study aimed to isolate, sequence, and analyze the COX2 DNA from individuals kept at the Giant Panda Protection and Research Center, China, and compare these sequences with those of the other Ursidae family members. Multiple sequence alignment showed that the COX2 gene had three point mutations that defined three haplotypes, with 60% of the sequences corresponding to haplotype I. The neutrality tests revealed that the COX2 gene was conserved throughout evolution, and the maximum likelihood phylogenetic analysis, using homologous sequences from other Ursidae species, showed clustering of the COX2 sequences of giant pandas, suggesting that this gene evolved differently in them.


Pilot analysis of the Motivation Assessment for Team Readiness, Integration, and Collaboration (MATRICx) using Rasch analysis.
Healthcare services and the production of healthcare knowledge are increasingly dependent on highly functioning, multidisciplinary teams, requiring greater awareness of individuals' readiness to collaborate in translational science teams. Yet, there is no comprehensive tool of individual motivations and threats to collaboration that can guide preparation of individuals for work on well-functioning teams. This prospective pilot study evaluated the preliminary psychometric properties of the Motivation Assessment for Team Readiness, Integration, and Collaboration (MATRICx). We examined 55 items of the MATRICx in a sample of 125 faculty, students and researchers, using contemporary psychometric methods (Rasch analysis). We found that the motivator and threat items formed separate constructs relative to collaboration readiness. Further, respondents who identified themselves as inexperienced at working on collaborative projects defined the motivation construct differently from experienced respondents. These results are consistent with differences in strategic alliances described in the literature-for example, inexperienced respondents reflected features of cooperation and coordination, such as concern with sharing information and compatibility of goals. In contrast, the more experienced respondents were concerned with issues that reflected a collective purpose, more typical of collaborative alliances. While these different types of alliances are usually described as representing varying aspects along a continuum, our findings suggest that collaboration might be better thought of as a qualitatively different state than cooperation or coordination. These results need to be replicated in larger samples, but the findings have implications for the development and design of educational interventions that aim to ready scientists and clinicians for greater interdisciplinary work.


Barriers and facilitators to adherence to secondary stroke prevention medications after stroke: analysis of survivors and caregivers views from an online stroke forum.
To identify barriers and facilitators of medication adherence in patients with stroke along with their caregivers. Qualitative thematic analysis of posts about secondary prevention medications, informed by Perceptions and Practicalities Approach. Posts written by the UK stroke survivors and their family members taking part in the online forum of the Stroke Association, between 2004 and 2011. 84 participants: 49 stroke survivors, 33 caregivers, 2 not stated, identified using the keywords 'taking medication', 'pills', 'size', 'side-effects', 'routine', 'blister' as well as secondary prevention medication terms. Perceptions reducing the motivation to adhere included dealing with medication side effects, questioning doctors' prescribing practices and negative publicity about medications, especially in regard to statins. Caregivers faced difficulties with ensuring medications were taken while respecting the patient's decisions not to take tablets. They struggled in their role as advocates of patient's needs with healthcare professionals. Not experiencing side effects, attributing importance to medications, positive personal experiences of taking tablets and obtaining modification of treatment to manage side effects were facilitators of adherence. Key practical barriers included difficulties with swallowing tablets, dealing with the burden of treatment and drug cost. Using medication storage devices, following routines and getting help with medications from caregivers were important facilitators of adherence. An online stroke forum is a novel and valuable resource to investigate use of secondary prevention medications. Analysis of this forum highlighted significant barriers and facilitators of medication adherence faced by stroke survivors and their caregivers. Addressing perceptual and practical barriers highlighted here can inform the development of future interventions aimed at improving adherence to secondary prevention medication after stroke.


Effect of ketone bodies on glucose production and utilization in the miniature pig.
The effect of ketone bodies on glucose production (Ra) and utilization (Rd) was investigated in the 24-h starved, conscious unrestrained miniature pig. Infusing Na-DL-beta-OH-butyrate (Na-DL-beta-OHB) and thus shifting the blood pH from 7.40 to 7.56 resulted in a decrease of Ra by 52% and of Rd by 45%, as determined by the isotope dilution technique. Simultaneously, the concentrations of arterial insulin and glucagon were slightly enhanced, whereas the plasma levels of glucose, lactate, pyruvate, alanine, alpha-amino-N, and free fatty acids (FFA) were all reduced. Infusion of Na-bicarbonate, which yielded a similar shift in blood pH, did not mimick these effects. Infusion of equimolar amounts of the ketoacid, yielding a blood pH of 7.35, induced similar metabolic alterations with respect to plasma glucose, Ra, Rd, and insulin; however, plasma alanine and alpha-amino-N increased. Infusing different amounts of Na-DL-beta-OHB resulting in plasma steady state levels of ketones from 0.25 to 1.5 mM had similar effects on arterial insulin and glucose kinetics. No dose dependency was observed. Prevention of the Na-DL-beta-OHB-induced hypoalaninemia by simultaneous infusion of alanine (1 mumol/kg X min) did not prevent hypoglycemia. Infusion of Na-DL-beta-OHB plus insulin (0.4 mU/kg X min) showed no additive effect on the inhibition of Ra. Ketones did not inhibit the insulin-stimulated metabolic clearance rate (MCR) for glucose. Infusion of somatostatin (0.2 micrograms/kg X min) initially decreased plasma glucose, Ra, and Rd, which was followed by an increase in plasma glucose and Ra; however, on infusion of somatostatin plus Na-DL-beta-OHB, hypoglycemia and the reduced Ra were maintained. In the anaesthetized 24-h starved miniature pig, Na-DL-beta-OHB infusion decreased the hepatic exchange for glucose, lactate, and FFA, whereas the exchange for glycerol, alanine, and alpha-amino-N as well as liver perfusion rate were unaffected. Simultaneously, portal glucagon and insulin as well as hepatic insulin extraction rate were elevated. Leg exchange for glucose, lactate, glycerol, alanine, alpha-amino-N, and FFA were decreased, while ketone body utilization increased. Repeated infusion of Na-DL-beta-OHB at the fourth, fifth, and sixth day of starvation in the conscious, unrestrained mini-pig resulted in a significant drop in urinary nitrogen (N)-excretion. However, this effect was mimicked by infusing equimolar amounts of Na-bicarbonate. In contrast, when only the ketoacid was given, urinary N-excretion accelerated. To summarize: (a) Ketone bodies decrease endogenous glucose production via an insulin-dependent mechanism; in addition, ketones probably exert a direct inhibitory action on gluconeogenesis. The ketone body-induced hypoalaninemia does not contribute to this effect. (b) The counterregulatory response to hypoglycemia is reduced by ketones. (c) As a consequence of the decrease in R(a), glucose utilization declines during ketone infusion. (d)The insulin-stimulated MCR for glucose is not affected by ketones. (e) Ketones in their physiological moiety do not show a protein-sparing effect.


Youth's narratives about family members smoking: parenting the parent- it's not fair!
Successful cancer prevention policies and programming for youth must be based on a solid understanding of youth's conceptualization of cancer and cancer prevention. Accordingly, a qualitative study examining youth's perspectives of cancer and its prevention was undertaken. Not surprisingly, smoking (i.e., tobacco cigarette smoking) was one of the dominant lines of discourse in the youth's narratives. This paper reports findings of how youth conceptualize smoking with attention to their perspectives on parental and family-related smoking issues and experiences. Seventy-five Canadian youth ranging in age from 11-19 years participated in the study. Six of the 75 youth had a history of smoking and 29 had parents with a history of smoking. Youth were involved in traditional ethnographic methods of interviewing and photovoice. Data analysis involved multiple levels of analysis congruent with ethnography. Youth's perspectives of parents and other family members' cigarette smoking around them was salient as represented by the theme: It's not fair. Youth struggled to make sense of why parents would smoke around their children and perceived their smoking as an unjust act. The theme was supported by four subthemes: 1) parenting the parent about the dangers of smoking; 2) the good/bad parent; 3) distancing family relationships; and 4) the prisoner. Instead of being talked to about smoking it was more common for youth to share stories of talking to their parents about the dangers of smoking. Parents who did not smoke were seen by youth as the good parent, as opposed to the bad parent who smoked. Smoking was an agent that altered relationships with parents and other family members. Youth who lived in homes where they were exposed to cigarette smoke felt like a trapped prisoner. Further research is needed to investigate youth's perceptions about parental cigarette smoking as well as possible linkages between youth exposed to second hand smoke in their home environment and emotional and lifestyle-related health difficulties. Results emphasize the relational impact of smoking when developing anti-tobacco and cancer prevention campaigns. Recognizing the potential toll that second-hand smoke can have on youth's emotional well-being, health care professionals are encouraged to give youth positive messages in coping with their parents' smoking behaviour.


Rapid and Specific Diagnosis of Extrapulmonary Tuberculosis by Immunostaining of Tissues and Aspirates With Anti-MPT64.
Extrapulmonary tuberculosis (EPTB) constitutes about 15% to 20% of all cases of tuberculosis (TB). The confirmation of EPTB has always been a challenge to laboratory personnel. We aim to evaluate the diagnostic potential of immunostaining with anti-MPT64 in various EPTB specimens. We studied a total of 51 TB cases and 38 non-TB control specimens comprising of fine-needle aspirates and formalin-fixed biopsies. These were investigated using a combination of the Ziehl-Neelsen method, the Lowenstein-Jensen culture, immunostaining with anti-MPT64 and anti-BCG, and nested-polymerase chain reaction (PCR) for IS6110. Results of all the tests were compared using nested-PCR as the gold standard. Diagnostic validation of immunostaining for anti-MPT64 was performed using nested-PCR as the gold standard. The overall sensitivity, specificity, and positive and negative predictive values for immunostaining with anti-MPT64 were 100%, 97%, 97%, and 100%, respectively. Immunostaining using anti-MPT64 is a rapid and sensitive method for establishing an early and specific diagnosis of Mycobacterium tuberculosis infection. The technique is simple to be incorporated into routine pathology laboratories.


Defects in mitochondrial dynamics and metabolomic signatures of evolving energetic stress in mouse models of familial Alzheimer's disease.
The identification of early mechanisms underlying Alzheimer's Disease (AD) and associated biomarkers could advance development of new therapies and improve monitoring and predicting of AD progression. Mitochondrial dysfunction has been suggested to underlie AD pathophysiology, however, no comprehensive study exists that evaluates the effect of different familial AD (FAD) mutations on mitochondrial function, dynamics, and brain energetics. We characterized early mitochondrial dysfunction and metabolomic signatures of energetic stress in three commonly used transgenic mouse models of FAD. Assessment of mitochondrial motility, distribution, dynamics, morphology, and metabolomic profiling revealed the specific effect of each FAD mutation on the development of mitochondrial stress and dysfunction. Inhibition of mitochondrial trafficking was characteristic for embryonic neurons from mice expressing mutant human presenilin 1, PS1(M146L) and the double mutation of human amyloid precursor protein APP(Tg2576) and PS1(M146L) contributing to the increased susceptibility of neurons to excitotoxic cell death. Significant changes in mitochondrial morphology were detected in APP and APP/PS1 mice. All three FAD models demonstrated a loss of the integrity of synaptic mitochondria and energy production. Metabolomic profiling revealed mutation-specific changes in the levels of metabolites reflecting altered energy metabolism and mitochondrial dysfunction in brains of FAD mice. Metabolic biomarkers adequately reflected gender differences similar to that reported for AD patients and correlated well with the biomarkers currently used for diagnosis in humans. Mutation-specific alterations in mitochondrial dynamics, morphology and function in FAD mice occurred prior to the onset of memory and neurological phenotype and before the formation of amyloid deposits. Metabolomic signatures of mitochondrial stress and altered energy metabolism indicated alterations in nucleotide, Krebs cycle, energy transfer, carbohydrate, neurotransmitter, and amino acid metabolic pathways. Mitochondrial dysfunction, therefore, is an underlying event in AD progression, and FAD mouse models provide valuable tools to study early molecular mechanisms implicated in AD.


Strouhal numbers and optimization of swimming by odontocete cetaceans.
Swimming efficiencies of fish and cetaceans have been related to a certain synchrony between stroke cycle frequency, peak-to-peak tail/fluke amplitude and mean swimming speed. These kinematic parameters form a non-dimensional wake parameter, referred to as a Strouhal number, which for the range between 0.20 and 0.40 has been associated with enhanced swimming efficiency for fish and cetaceans. Yet to date there has been no direct experimental substantiation of what Strouhal numbers are preferred by swimming cetaceans. To address this lack of data, a total of 248 Strouhal numbers were calculated for the captive odontocete cetaceans Tursiops truncatus, Pseudorca crassidens, Orcinus orca, Globicephala melaena, Lagenorhynchus obliquidens and Stenella frontalis. Although the average Strouhal number calculated for each species is within the accepted range, considerable scatter is found in the data both within species and among individuals. A greater proportion of Strouhal values occur between 0.20 and 0.30 (74%) than the 0.25-0.35 (55%) range predicted for maximum swimming efficiency. Within 0.05 Strouhal increments, the greatest number of Strouhal values was found between 0.225 and 0.275 (44%). Where propulsive efficiency data were available (Tursiops truncatus, Pseudorca crassidens, Orcinus orca), peak swimming efficiency corresponded to this same Strouhal range. The odontocete cetacean data show that, besides being generally limited to a range of Strouhal numbers between 0.20 and 0.40, the kinematic parameters comprising the Strouhal number provide additional constraints. Fluke-beat frequency normalized by the ratio of swimming speed to body length was generally restricted from 1 to 2, whereas peak-to-peak fluke amplitude normalized by body length occurred predominantly between 0.15 and 0.25. The results indicate that the kinematics of the propulsive flukes of odontocete cetaceans are not solely dependent on Strouhal number, and the Strouhal number range for odontocete cetaceans occurs at slightly ( approximately 20%) lower values than previously predicted for maximum swimming efficiency.


Influence of tomato genotype on growth of inoculated and indigenous bacteria in the spermosphere.
We previously demonstrated a genetic basis in tomato for support of the growth of a biological control agent, Bacillus cereus UW85, in the spermosphere after seed inoculation (K. P. Smith, J. Handelsman, and R. M. Goodman, Proc. Natl. Acad. Sci. USA 96:4786-4790, 1999). Here we report results of studies examining the host effect on the support of growth of Bacillus and Pseudomonas strains, both inoculated on seeds and recruited from soil, using selected inbred tomato lines from the recombinant inbred line (RIL) population used in our previous study. Two tomato lines, one previously found to support high and the other low growth of B. cereus UW85 in the spermosphere, had similar effects on growth of each of a diverse, worldwide collection of 24 B. cereus strains that were inoculated on seeds and planted in sterilized vermiculite. In contrast, among RILs that differed for support of B. cereus UW85 growth in the spermosphere, we found no difference for support of growth of the biocontrol strains Pseudomonas fluorescens 2-79 or Pseudomonas aureofaciens AB254. Thus, while the host effect on growth extended to all strains of B. cereus examined, it was not exerted on other bacterial species tested. When seeds were inoculated with a marked mutant of B. cereus UW85 and planted in soil, RIL-dependent high and low support of bacterial growth was observed that was similar to results from experiments conducted in sterilized vermiculite. When uninoculated seeds from two of these RILs were planted in soil, changes in population levels of indigenous Bacillus and fluorescent Pseudomonas bacteria differed, as measured over time by culturing and direct microscopy, from growth patterns observed in the inoculation experiments. Neither RIL supported detectable levels of growth of indigenous Bacillus soil bacteria, while the line that supported growth of inoculated B. cereus UW85 supported higher growth of indigenous fluorescent pseudomonads and total bacteria. The vermiculite system used in these experiments was predictive for growth of B. cereus UW85 inoculated on seeds and grown in soil, but the patterns of growth of inoculated strains-both Bacillus and Pseudomonas spp.-did not reflect host genotype effects on indigenous microflora recruited from soil to the spermosphere.


Variation in secondary metabolites in a unique set of tomato accessions collected in Turkey.
In this study, 50 tomato landraces grown in Turkey were investigated in terms of their secondary metabolite profiles. Each accession was planted in 2016 and 2017 in 3 replicates in an open field. In this study, color, pH and brix of the fruit samples were measured and an unbiased LCMS-based metabolomics approach was applied. Based on Principal Components Analysis (PCA) and Hierarchical Cluster Analysis (HCA) of the relative abundance levels of >250 metabolites, it could be concluded that fruit size was the most influential to the biochemical composition, rather than the geographical origin of accessions. Results indicated substantial biodiversity in various metabolites generally regarded as key to fruit quality aspects, including sugars; phenolic compounds like phenylpropanoids and flavonoids; alkaloids and glycosides of flavour-related volatile compounds. The phytochemical data provides insight into which Turkish accessions might be most promising as starting materials for the tomato processing and breeding industries.


Neurotransmitter identity is acquired in a lineage-restricted manner in the Drosophila CNS.
The vast majority of the adult fly ventral nerve cord is composed of 34 hemilineages, which are clusters of lineally related neurons. Neurons in these hemilineages use one of the three fast-acting neurotransmitters (acetylcholine, GABA, or glutamate) for communication. We generated a comprehensive neurotransmitter usage map for the entire ventral nerve cord. We did not find any cases of neurons using more than one neurotransmitter, but found that the acetylcholine specific gene ChAT is transcribed in many glutamatergic and GABAergic neurons, but these transcripts typically do not leave the nucleus and are not translated. Importantly, our work uncovered a simple rule: All neurons within a hemilineage use the same neurotransmitter. Thus, neurotransmitter identity is acquired at the stem cell level. Our detailed transmitter- usage/lineage identity map will be a great resource for studying the developmental basis of behavior and deciphering how neuronal circuits function to regulate behavior.


RISUS study: Rugby Injury Surveillance in Ulster Schools.
To examine injury patterns in adolescent rugby players and determine factors associated with injury risk. Prospective injury surveillance study. N=28 Grammar Schools in Ulster, Ireland (2014-2015 playing season). 825 adolescent rugby players, across in 28 school first XV rugby squads; mean age 16.9 years. Injuries were classified by body part and diagnosis, and injury incidence using injuries per 1000 match hours of exposure. HRs for injury were calculated through Cox proportional hazard regression after correction for influential covariates. A total of n=426 injuries were reported across the playing season. Over 50% of injuries occurred in the tackle situation or during collisions (270/426), with few reported during set plays. The 3 most common injury sites were head/face (n=102, 23.9%), clavicle/shoulder (n=65, 15.3%) and the knee (n=56, 13.1%). Sprain (n=133, 31.2%), concussion (n=81, 19%) and muscle injury (n=65, 15.3%) were the most common diagnoses. Injury incidence is calculated at 29.06 injuries per 1000 match hours. There were no catastrophic injuries. A large percentage of injuries (208/424) resulted in absence from play for more than 28 days. Concussion carried the most significant time out from play (n=33; 15.9%), followed by dislocations of the shoulder (n=22; 10.6%), knee sprains (n=19, 9.1%), ankle sprains (n=14, 6.7%), hand/finger/thumb (n=11; 5.3%). 36.8% of participants in the study (304/825) suffered at least one injury during the playing season. Multivariate models found higher risk of injury (adjusted HR (AHR); 95% CI) with: higher age (AHR 1.45; 1.14 to 1.83), heavier weight (AHR 1.32; 1.04 to 1.69), playing representative rugby (AHR 1.42; 1.06 to 1.90) and undertaking regular strength training (AHR 1.65; 1.11 to 2.46). Playing for a lower ranked team (AHR 0.67; 0.49 to 0.90) and wearing a mouthguard (AHR 0.70; 0.54 to 0.92) were associated with lower risk of injury. There was a high incidence of severe injuries, with concussion, ankle and knee ligament injuries and upper limb fractures/dislocations causing greatest time loss. Players were compliant with current graduated return-to-play regulations following concussion. Physical stature and levels of competition were important risk factors and there was limited evidence for protective equipment.


Ethylene receptors function as components of high-molecular-mass protein complexes in Arabidopsis.
The gaseous plant hormone ethylene is perceived in Arabidopsis thaliana by a five-member receptor family composed of ETR1, ERS1, ETR2, ERS2, and EIN4. Gel-filtration analysis of ethylene receptors solubilized from Arabidopsis membranes demonstrates that the receptors exist as components of high-molecular-mass protein complexes. The ERS1 protein complex exhibits an ethylene-induced change in size consistent with ligand-mediated nucleation of protein-protein interactions. Deletion analysis supports the participation of multiple domains from ETR1 in formation of the protein complex, and also demonstrates that targeting to and retention of ETR1 at the endoplasmic reticulum only requires the first 147 amino acids of the receptor. A role for disulfide bonds in stabilizing the ETR1 protein complex was demonstrated by use of reducing agents and mutation of Cys4 and Cys6 of ETR1. Expression and analysis of ETR1 in a transgenic yeast system demonstrates the importance of Cys4 and Cys6 of ETR1 in stabilizing the receptor for ethylene binding. These data support the participation of ethylene receptors in obligate as well as ligand-dependent non-obligate protein interactions. These data also suggest that different protein complexes may allow for tailoring of the ethylene signal to specific cellular environments and responses.


The use of navigation systems in naturalistic driving.
In this study, we assessed the use of portable navigation systems in everyday driving by applying in-vehicle naturalistic driving. Experienced users of navigation systems, 7 females and 14 males, were provided with a specially equipped vehicle for approximately 1 month. Their trips were recorded using 4 cameras, Global Positioning System (GPS) data, and other sensor data. The drivers' navigation system use data were coded from the video recordings, which showed how often and for how long the system was activated and how often and for how long a driver operated the system. The system was activated for 23% of trips, predominantly on longer and unique trips. Analyses of the percentage of time for which the speed limit was exceeded showed no evidence of differences between trips for which the navigation system was used or not used. On trips for which the navigation system was activated, participants spent about 5% of trip time interacting with the device. About 40% of interacting behavior took place in the first 10% of the trip time, and about 35% took place while the car was standing still or moving at a very low speed; that is, 0-10 km/h. These results shed light on how and when drivers use navigation systems. They suggest that although drivers regulate their use of such systems to some extent, they often perform risky tasks while driving.


Primary humoral antibody response to Coxiella burnetii, the causative agent of Q fever.
Of 147 patients with acute Q fever diagnosed during a major outbreak in Birmingham, England, in early summer 1989, 41 provided sets of sera which allowed us to make a detailed analysis of the primary humoral immune response. Antibody titers specific for Coxiella burnetii were measured by the complement fixation test and by an immunoglobulin M (IgM)- and IgG-specific indirect immunofluorescence test. The relative avidity of specific IgGs was determined by the indirect immunofluorescence test with and without treatment of antigen-antibody complexes with 8 M urea. The IgG subclass responses after primary infection and their avidities were also determined for a limited number of paired serum specimens. Specific IgM titers persisted for more than 6 months in the majority of cases and were therefore not a sufficient criterion for the diagnosis of recent infection. However, for serial samples the antibody titer ratios (IgG/IgM) and the ratios (IgG titer with treatment/IgG titer without treatment) that indicated relative avidity changed significantly, depending on the time postinfection. Within the IgG class, the C. burnetii-specific antibody response over time was almost exclusively represented by subclass 1 molecules, which thus showed affinity maturation.


Detection of neuritic plaques in Alzheimer's disease by magnetic resonance microscopy.
Magnetic resonance microscopy (MRM) theoretically provides the spatial resolution and signal-to-noise ratio needed to resolve neuritic plaques, the neuropathological hallmark of Alzheimer's disease (AD). Two previously unexplored MR contrast parameters, T2* and diffusion, are tested for plaque-specific contrast to noise. Autopsy specimens from nondemented controls (n = 3) and patients with AD (n = 5) were used. Three-dimensional T2* and diffusion MR images with voxel sizes ranging from 3 x 10(-3) mm(3) to 5.9 x 10(-5) mm(3) were acquired. After imaging, specimens were cut and stained with a microwave king silver stain to demonstrate neuritic plaques. From controls, the alveus, fimbria, pyramidal cell layer, hippocampal sulcus, and granule cell layer were detected by either T2* or diffusion contrast. These structures were used as landmarks when correlating MRMs with histological sections. At a voxel resolution of 5.9 x 10(-5) mm(3), neuritic plaques could be detected by T2*. The neuritic plaques emerged as black, spherical elements on T2* MRMs and could be distinguished from vessels only in cross-section when presented in three dimension. Here we provide MR images of neuritic plaques in vitro. The MRM results reported provide a new direction for applying this technology in vivo. Clearly, the ability to detect and follow the early progression of amyloid-positive brain lesions will greatly aid and simplify the many possibilities to intervene pharmacologically in AD.


Overexpression of microRNA-205-5p promotes cholangiocarcinoma growth by reducing expression of homeodomain-interacting protein kinase 3.
The microRNA miR-205-5p has diverse effects in different malignancies, including cholangiocarcinoma (CCA), but its effects on CCA progression is unclear. Here we investigated the role and function of miR-205-5p in CCA. Three CCA cell lines and human serum samples were found to have much higher expression levels of miR-205-5p than seen in typical cholangiocyte cell lines and healthy controls. Inhibition of miR-205-5p suppressed CCA cell motility, invasion and proliferation of KKU-213B whereby overexpression of miR-205-5p promoted cell proliferation and motility of KKU-100 cells. Bioinformatics tools (miRDB, TargetScan, miRWalk, and GEPIA) all predicted various miR-205-5p targets. Experiments using miR-205-5p inhibitor and mimic indicated that homeodomain-interacting protein kinase 3 (HIPK3) was a potential direct target of miR-205-5p. Overexpression of HIPK3 using HIPK3 plasmid cloning DNA suppressed migration and proliferation of KKU-100 cells. Notably, HIPK3 expression was lower in human CCA tissues than in normal adjacent tissues. High HIPK3 expression was significantly associated with longer survival time of CCA patients. Multivariate regression analysis indicated tissue HIPK3 levels as an independent prognostic factor for CCA patients. These findings indicate that overexpression of miR-205-5p promotes CCA cells proliferation and migration partly via HIPK3-dependent way. Therefore, targeting miR-205-5p may be a potential treatment approach for CCA.


High repetition rate femtosecond double pass optical parametric generator with more than 2 W tunable output in the NIR.
We demonstrate a highly efficient double pass optical parametric generator based on periodically poled MgO-doped congruent LiNbO3. More than two watts of tunable near-IR radiation (1370-1650 nm) are generated by directly pumping the system with 550 fs pulses from a 42 MHz repetition rate passively mode-locked Yb:KGW oscillator. Pulse durations below 200 fs were achieved without further compression techniques. The system is extremely efficient, compact, cost effective, easy to align and easy to operate, which makes it an interesting alternative to more complex optical parametric oscillators or optical parametric amplifiers.


The dynamics of embodiment: a field theory of infant perseverative reaching.
The overall goal of this target article is to demonstrate a mechanism for an embodied cognition. The particular vehicle is a much-studied, but still widely debated phenomenon seen in 7-12 month-old-infants. In Piaget's classic "A-not-B error," infants who have successfully uncovered a toy at location "A" continue to reach to that location even after they watch the toy hidden in a nearby location "B." Here, we question the traditional explanations of the error as an indicator of infants' concepts of objects or other static mental structures. Instead, we demonstrate that the A-not-B error and its previously puzzling contextual variations can be understood by the coupled dynamics of the ordinary processes of goal-directed actions: looking, planning, reaching, and remembering. We offer a formal dynamic theory and model based on cognitive embodiment that both simulates the known A-not-B effects and offers novel predictions that match new experimental results. The demonstration supports an embodied view by casting the mental events involved in perception, planning, deciding, and remembering in the same analogic dynamic language as that used to describe bodily movement, so that they may be continuously meshed. We maintain that this mesh is a pre-eminently cognitive act of "knowing" not only in infancy but also in everyday activities throughout the life span.


Hyperplasia of mantle/marginal zone B cells with clear cytoplasm in peripheral lymph nodes. A clinicopathologic study of 35 cases.
We describe 35 peripheral lymph nodes classified as mantle cell/marginal zone B-cell hyperplasia with clear cells using morphologic and immunologic findings. For the purpose of this study, we obtained clinical follow-up information and performed immunoglobulin gene rearrangement studies on paraffin sections by polymerase chain reaction. Architecturally, the nodes were suggestive of a benign process: no pericapsular infiltration, sinuses readily identified, scattered reactive follicles present, and paracortical nodular hyperplasia present. No monocytoid B cells were present. Focally, small lymphoid cells with round nuclei and clear cytoplasm (clear cells) formed monomorphic nodular, inverse follicular, and/or marginal zone patterns. Flow cytometry and immunohistochemical analysis revealed neither light chain restriction nor an aberrant B-cell phenotype. Immunoglobulin gene rearrangement studies showed a clonal band in 1 of 26 cases in which DNA was amplified. To ascertain the clinical relevance of this positive case, follow-up information was obtained 30 months after the initial biopsy; the 83-year-old woman was alive without treatment but had splenomegaly and bone marrow involvement by marginal zone B-cell lymphoma. The morphologic and immunologic criteria used for diagnosis of mantle cell/marginal zone B-cell hyperplasia with clear cytoplasm are valid; however, to rule out the possibility of occult lymphoma, immunoglobulin gene rearrangement studies and clinical follow-up are necessary.