Transistor memory devices with large memory windows, using multi-stacking of densely packed, hydrophobic charge trapping metal nanoparticle array.
Organic field-effect transistor (OFET) memories have rapidly evolved from low-cost and flexible electronics with relatively low-memory capacities to memory devices that require high-capacity memory such as smart memory cards or solid-state hard drives. Here, we report the high-capacity OFET memories based on the multilayer stacking of densely packed hydrophobic metal NP layers in place of the traditional transistor memory systems based on a single charge trapping layer. We demonstrated that the memory performances of devices could be significantly enhanced by controlling the adsorption isotherm behavior, multilayer stacking structure and hydrophobicity of the metal NPs. For this study, tetraoctylammonium (TOA)-stabilized Au nanoparticles (TOA-Au(NPs)) were consecutively layer-by-layer (LbL) assembled with an amine-functionalized poly(amidoamine) dendrimer (PAD). The formed (PAD/TOA-Au(NP))(n) films were used as a multilayer stacked charge trapping layer at the interface between the tunneling dielectric layer and the SiO2 gate dielectric layer. For a single AuNP layer (i.e. PAD/TOA-Au(NP))1) with a number density of 1.82 × 10(12) cm(-2), the memory window of the OFET memory device was measured to be approximately 97 V. The multilayer stacked OFET memory devices prepared with four Au(NP) layers exhibited excellent programmable memory properties (i.e. a large memory window (ΔV(th)) exceeding 145 V, a fast switching speed (1 μs), a high program/erase (P/E) current ratio (greater than 10(6)) and good electrical reliability) during writing and erasing over a relatively short time scale under an operation voltage of 100 V applied at the gate.


Diet in Irritable Bowel Syndrome (IBS): Interaction with Gut Microbiota and Gut Hormones.
Diet plays an important role not only in the pathophysiology of irritable bowel syndrome (IBS), but also as a tool that improves symptoms and quality of life. The effects of diet seem to be a result of an interaction with the gut bacteria and the gut endocrine cells. The density of gut endocrine cells is low in IBS patients, and it is believed that this abnormality is the direct cause of the symptoms seen in IBS patients. The low density of gut endocrine cells is probably caused by a low number of stem cells and low differentiation progeny toward endocrine cells. A low fermentable oligo-, di-, monosaccharide, and polyol (FODMAP) diet and fecal microbiota transplantation (FMT) restore the gut endocrine cells to the level of healthy subjects. It has been suggested that our diet acts as a prebiotic that favors the growth of a certain types of bacteria. Diet also acts as a substrate for gut bacteria fermentation, which results in several by-products. These by-products might act on the stem cells in such a way that the gut stem cells decrease, and consequently, endocrine cell numbers decrease. Changing to a low-FODMAP diet or changing the gut bacteria through FMT improves IBS symptoms and restores the density of endocrine cells.


1,4-Dihydropyridine binding sites in moss plasma membranes. Properties of receptors for a calcium channel antagonist.
An increase in cytoplasmic calcium is an early event in hormone (cytokinin)-induced vegetative bud formation in the moss Physcomitrella patens. Whole cell and calcium transport studies have implicated 1,4-dihydropyridine-sensitive calcium channels in this increase in cellular calcium. To understand the molecular nature of the dihydropyridine-sensitive calcium channel, we have established conditions for the binding of the arylazide 1,4-dihydropyridine, [3H]azidopine, to its receptor in moss plasma membranes. [3H]Azidopine bound specifically in a saturable and reversible manner. The KD for [3H]azidopine binding was 5.2 nM and the Bmax was 35.6 pmol/mg of protein. Association and dissociation of the receptor and [3H]azidopine were temperature-dependent, and association varied as a function of pH. Binding was inhibited by dihydropyridine, phenylalkylamine, and benzothiazepine calcium channel blockers, bepridil, lanthanum, and N-ethylmaleimide. [3H]Azidopine binding was stimulated by cations including calcium, strontium, manganese, and barium. [3H]Azidopine binding was also stimulated by cytokinin with a Km value for kinetin of 0.13 nM. These studies utilize a simple plant system to provide a biochemical framework for understanding calcium regulation during development and have implications for understanding mechanisms of signal transduction in plants.


National narcolepsy survey.
Narcolepsy is characterised by excessive daytime sleepiness and cataplexy and has a prevalence of 25 per 100,000. We suspect this is higher than presently seen in the Republic of Ireland. We aimed to calculate the Irish prevalence of Narcolepsy and to examine current management practices. We conducted an online survey of respiratory physicians, neurologists, paediatric neurologists, and psychiatrists with an interest in sleep disorders (73% response rate). Of this group, a total of 16 physicians managed 180 patients prior to January 2009. A clinical diagnosis alone was reached in 67 (41%) patients, the remainder by polysomnography or multiple sleep latency testing. No patients were diagnosed by cerebro-spinal fluid analysis of hypocretin levels. While 70 (42%) patients received modafanil, only 7 (4%) were treated with sodium oxybate. Even allowing for missing data it is apparent that Narcolepsy is hugely under-diagnosed in Ireland, however, current practises adhere with new international guidelines.


Rhesus TRIM5α disrupts the HIV-1 capsid at the inter-hexamer interfaces.
TRIM proteins play important roles in the innate immune defense against retroviral infection, including human immunodeficiency virus type-1 (HIV-1). Rhesus macaque TRIM5α (TRIM5α(rh)) targets the HIV-1 capsid and blocks infection at an early post-entry stage, prior to reverse transcription. Studies have shown that binding of TRIM5α to the assembled capsid is essential for restriction and requires the coiled-coil and B30.2/SPRY domains, but the molecular mechanism of restriction is not fully understood. In this study, we investigated, by cryoEM combined with mutagenesis and chemical cross-linking, the direct interactions between HIV-1 capsid protein (CA) assemblies and purified TRIM5α(rh) containing coiled-coil and SPRY domains (CC-SPRY(rh)). Concentration-dependent binding of CC-SPRY(rh) to CA assemblies was observed, while under equivalent conditions the human protein did not bind. Importantly, CC-SPRY(rh), but not its human counterpart, disrupted CA tubes in a non-random fashion, releasing fragments of protofilaments consisting of CA hexamers without dissociation into monomers. Furthermore, such structural destruction was prevented by inter-hexamer crosslinking using P207C/T216C mutant CA with disulfide bonds at the CTD-CTD trimer interface of capsid assemblies, but not by intra-hexamer crosslinking via A14C/E45C at the NTD-NTD interface. The same disruption effect by TRIM5α(rh) on the inter-hexamer interfaces also occurred with purified intact HIV-1 cores. These results provide insights concerning how TRIM5α disrupts the virion core and demonstrate that structural damage of the viral capsid by TRIM5α is likely one of the important components of the mechanism of TRIM5α-mediated HIV-1 restriction.


The Anopheles dirus complex: spatial distribution and environmental drivers.
The Anopheles dirus complex includes efficient malaria vectors of the Asian forested zone. Studies suggest ecological and biological differences between the species of the complex but variations within species suggest possible environmental influences. Behavioural variation might determine vector capacity and adaptation to changing environment. It is thus necessary to clarify the species distributions and the influences of environment on behavioural heterogeneity. A literature review highlights variation between species, influences of environmental drivers, and consequences on vector status and control. The localisation of collection sites from the literature and from a recent project (MALVECASIA) produces detailed species distributions maps. These facilitate species identification and analysis of environmental influences. The maps give a good overview of species distributions. If species status partly explains behavioural heterogeneity, occurrence and vectorial status, some environmental drivers have at least the same importance. Those include rainfall, temperature, humidity, shade, soil type, water chemistry and moon phase. Most factors are probably constantly favourable in forest. Biological specificities, behaviour and high human-vector contact in the forest can explain the association of this complex with high malaria prevalence, multi-drug resistant Plasmodium falciparum and partial control failure of forest malaria in Southeast Asia. Environmental and human factors seem better than species specificities at explaining behavioural heterogeneity. Although forest seems essential for mosquito survival, adaptations to orchards and wells have been recorded. Understanding the relationship between landscape components and mosquito population is a priority in foreseeing the influence of land-cover changes on malaria occurrence and in shaping control strategies for the future.


AFM mapping of the elastic properties of brain tissue reveals kPa μm(-1) gradients of rigidity.
It is now well established that the mechanical environment of the cells in tissues deeply impacts cellular fate, including life cycle, differentiation and tumor progression. Designs of biomaterials already include the control of mechanical parameters, and in general, their main focus is to control the rheological properties of the biomaterials at a macroscopic scale. However, recent studies have demonstrated that cells can stress their environment below the micron scale, and therefore could possibly respond to the rheological properties of their environment at this micron scale. In this context, probing the mechanical properties of physiological cellular environments at subcellular scales is becoming critical. To this aim, we performed in vitro indentation measurements using AFM on sliced human pituitary gland tissues. A robust methodology was implemented using elasto-adhesive models, which shows that accounting for the adhesion of the probe on the tissue is critical for the reliability of the measurement. In addition to quantifying for the first time the rigidity of normal pituitary gland tissue, with a geometric mean of 9.5 kPa, our measurements demonstrated that the mechanical properties of this tissue are far from uniform at subcellular scales. Gradients of rigidity as large as 12 kPa μm(-1) were observed. This observation suggests that physiological rigidity can be highly non-uniform at the micron-scale.


Prospective relations between family conflict and adolescent maladjustment: security in the family system as a mediating process.
Conflict in specific family systems (e.g., interparental, parent-child) has been implicated in the development of a host of adjustment problems in adolescence, but little is known about the impact of family conflict involving multiple family systems. Furthermore, questions remain about the effects of family conflict on symptoms of specific disorders and adjustment problems and the processes mediating these effects. The present study prospectively examines the impact of family conflict and emotional security about the family system on adolescent symptoms of specific disorders and adjustment problems, including the development of symptoms of anxiety, depression, conduct problems, and peer problems. Security in the family system was examined as a mediator of these relations. Participants included 295 mother-father-adolescent families (149 girls) participating across three annual time points (grades 7-9). Including auto-regressive controls for initial levels of emotional insecurity and multiple adjustment problems (T1), higher-order emotional insecurity about the family system (T2) mediated relations between T1 family conflict and T3 peer problems, anxiety, and depressive symptoms. Further analyses supported specific patterns of emotional security/insecurity (i.e., security, disengagement, preoccupation) as mediators between family conflict and specific domains of adolescent adjustment. Family conflict was thus found to prospectively predict the development of symptoms of multiple specific adjustment problems, including symptoms of depression, anxiety, conduct problems, and peer problems, by elevating in in adolescent's emotional insecurity about the family system. The clinical implications of these findings are considered.


Molecular evidence of an interaction between prenatal environmental exposures and birth outcomes in a multiethnic population.
Inner-city, minority populations are high-risk groups for adverse birth outcomes and also are more likely to be exposed to environmental contaminants, including environmental tobacco smoke (ETS), benzo[a]pyrene (BaP), and other polycyclic aromatic hydrocarbons (PAHs) found in urban air. In a sample of nonsmoking African-American and Dominican women, we evaluated the effects on birth outcomes of prenatal exposure to ETS, using questionnaire data and plasma cotinine as a biomarker of exposure, and environmental PAHs using BaP-DNA adducts as a molecular dosimeter. We previously reported that among African Americans, high prenatal exposure to PAHs estimated by prenatal personal air monitoring was associated with lower birth weight (p = 0.003) and smaller head circumference (p = 0.01) after adjusting for potential confounders. In the present analysis, self-reported ETS was associated with decreased head circumference (p = 0.04). BaP-DNA adducts were not correlated with ETS or dietary PAHs. There was no main effect of BaP-DNA adducts on birth outcomes. However, there was a significant interaction between the two pollutants such that the combined exposure to high ETS and high adducts had a significant multiplicative effect on birth weight (p = 0.04) and head circumference (p = 0.01) after adjusting for ethnicity, sex of newborns, maternal body mass index, dietary PAHs, and gestational age. This study provides evidence that combined exposure to environmental pollutants at levels currently encountered in New York City adversely affects fetal development.


Intrahepatic bile duct regeneration in mice does not require Hnf6 or Notch signaling through Rbpj.
The potential for intrahepatic bile duct (IHBD) regeneration in patients with bile duct insufficiency diseases is poorly understood. Notch signaling and Hnf6 have each been shown to be important for the morphogenesis of IHBDs in mice. One congenital pediatric liver disease characterized by reduced numbers of IHBDs, Alagille syndrome, is associated with mutations in Notch signaling components. Therefore, we investigated whether liver cell plasticity could contribute to IHBD regeneration in mice with disruptions in Notch signaling and Hnf6. We studied a mouse model of bile duct insufficiency with liver epithelial cell-specific deficiencies in Hnf6 and Rbpj, a mediator of canonical Notch signaling. Albumin-Cre Hnf6(flox/flox)Rbpj(flox/flox) mice initially developed no peripheral bile ducts. The evolving postnatal liver phenotype was analyzed using IHBD resin casting, immunostaining, and serum chemistry. With age, Albumin-Cre Hnf6(flox/flox)Rbpj(flox/flox) mice mounted a ductular reaction extending through the hepatic tissue and then regenerated communicating peripheral IHBD branches. Rbpj and Hnf6 were determined to remain absent from biliary epithelial cells constituting the ductular reaction and the regenerated peripheral IHBDs. We report the expression of Sox9, a marker of biliary epithelial cells, in cells expressing hepatocyte markers. Tissue analysis indicates that reactive ductules did not arise directly from preexisting hilar IHBDs. We conclude that liver cell plasticity is competent for regeneration of IHBDs independent of Notch signaling via Rbpj and Hnf6.


Beyond Consent: Building Trusting Relationships With Diverse Populations in Precision Medicine Research.
With the growth of precision medicine research on health data and biospecimens, research institutions will need to build and maintain long-term, trusting relationships with patient-participants. While trust is important for all research relationships, the longitudinal nature of precision medicine research raises particular challenges for facilitating trust when the specifics of future studies are unknown. Based on focus groups with racially and ethnically diverse patients, we describe several factors that influence patient trust and potential institutional approaches to building trustworthiness. Drawing on these findings, we suggest several considerations for research institutions seeking to cultivate long-term, trusting relationships with patients: (1) Address the role of history and experience on trust, (2) engage concerns about potential group harm, (3) address cultural values and communication barriers, and (4) integrate patient values and expectations into oversight and governance structures.


Setting a research agenda for interprofessional education and collaborative practice in the context of United States health system reform.
Interprofessional education (IPE) and collaborative practice (CP) have been prolific areas of inquiry exploring research questions mostly concerned with local program and project assessment. The actual sphere of influence of this research has been limited. Often discussed separately, this article places IPE and CP in the same conceptual space. The interface of these form a nexus where new knowledge creation may be facilitated. Rigorous research on IPE in relation to CP that is relevant to and framed by health system reform in the U.S. is the ultimate research goal of the National Center for Interprofessional Practice and Education at the University of Minnesota. This paper describes the direction and scope for a focused and purposive IPECP research agenda linked to improvement in health outcomes, contextualized by health care reform in the U.S. that has provided a revitalizing energy for this area of inquiry. A research agenda articulates a focus, meaningful and robust questions, and a theory of change within which intervention outcomes are examined. Further, a research agenda identifies the practices the area of inquiry is interested in informing, and the types of study designs and analytic approaches amenable to carrying out the proposed work.


Temperature- and excitation wavelength-dependent emission in a manganese(ii) complex.
A mononuclear manganese(ii) complex with a chelating 4-(3,5-diphenyl-1H-pyrazol-1-yl)-6-(piperidin-1-yl)pyrimidine ligand (L), [MnL2Cl2]·H2O, shows intriguing excitation wavelength-dependent emission. Depending on the excitation wavelength, the complex demonstrates three emission bands with the maxima at 380 nm, 440 nm and 495 nm. The 380 nm and 440 nm emissions originate from the π → π* and n → π* ligand-centered transitions. The long-wave 495 nm emission with microsecond lifetimes is related to the d-d transitions and/or metal-to-ligand and halogen-to-ligand charge transfer. The emission behavior of this complex is strongly temperature-dependent: upon cooling from 300 K down to 77 K, the intensity of emission considerably increases. The enhancement of the luminescence upon cooling is accompanied by the appearance of the vibrational structure. This complex is the first example of manganese(ii) complexes demonstrating excitation wavelength-dependent emission.


Divergent drivers of the microbial methane sink in temperate forest and grassland soils.
Aerated topsoils are important sinks for atmospheric methane (CH4 ) via oxidation by CH4 -oxidizing bacteria (MOB). However, intensified management of grasslands and forests may reduce the CH4 sink capacity of soils. We investigated the influence of grassland land-use intensity (150 sites) and forest management type (149 sites) on potential atmospheric CH4 oxidation rates (PMORs) and the abundance and diversity of MOB (with qPCR) in topsoils of three temperate regions in Germany. PMORs measurements in microcosms under defined conditions yielded approximately twice as much CH4 oxidation in forest than in grassland soils. High land-use intensity of grasslands had a negative effect on PMORs (-40%) in almost all regions and fertilization was the predominant factor of grassland land-use intensity leading to PMOR reduction by 20%. In contrast, forest management did not affect PMORs in forest soils. Upland soil cluster (USC)-α was the dominant group of MOBs in the forests. In contrast, USC-γ was absent in more than half of the forest soils but present in almost all grassland soils. USC-α abundance had a direct positive effect on PMOR in forest, while in grasslands USC-α and USC-γ abundance affected PMOR positively with a more pronounced contribution of USC-γ than USC-α. Soil bulk density negatively influenced PMOR in both forests and grasslands. We further found that the response of the PMORs to pH, soil texture, soil water holding capacity and organic carbon and nitrogen content differ between temperate forest and grassland soils. pH had no direct effects on PMOR, but indirect ones via the MOB abundances, showing a negative effect on USC-α, and a positive on USC-γ abundance. We conclude that reduction in grassland land-use intensity and afforestation has the potential to increase the CH4 sink function of soils and that different parameters determine the microbial methane sink in forest and grassland soils.


Systematic Review and Meta-Analysis to Establish the Association of Common Genetic Variations in Vitamin D Binding Protein With Chronic Obstructive Pulmonary Disease.
Background: Vitamin-D binding protein (DBP) also known as GC protein, is a major determinant for vitamin- D metabolism and transport. GC1F, GC1S, and GC2 are the three allelic variants (denoted as rs4588 and rs7041) of GC, and known to be associated with chronic obstructive pulmonary disease (COPD). However, contradictory reports and population specific risk attributed by these alleles warranted detailed genetic epidemiology study to establish the association between GC variants and COPD. In this study we performed a meta-analysis and investigated the genetic architecture of GC locus to establish the association and uncover the plausible reason for allelic heterogeneity. Methods: Published cross-sectional case control studies were screened and meta-analysis was performed between GC variants and COPD outcome. RevMan-v5.3 software was used to perform random and/or fixed models to calculate pooled odds ratio (Meta-OR). Linkage disequilibrium (LD) and haplotypes at GC locus were evaluated using 1000 Genomes genotype data. In silico functional implications of rs4588 and rs7041 was tested using publicly available tools. Results: GC1F allele and GC1F/1F genotype were found to confer COPD risk in overall meta-analysis. GC1S/1S was found to confer risk only among Europeans. In silico investigation of rs4588 and rs7041 identified strong eQTL effects and potential role in regulation of GC expression. Large differences in allele frequencies, linkage disequilibrium (LD) and haplotypes were identified at GC locus across different populations (Japanese, African, Europeans, and Indians), which may explain the variable association of different GC alleles in different populations. Conclusion: GC1F and GC1F/1F impose significant genetic risk for COPD, among Asians. Considerable differences in allele frequencies and LD structure in GC locus may impose population specific risk.


Functional Specificity and Sex Differences in the Neural Circuits Supporting the Inhibition of Automatic Imitation.
Humans show an involuntary tendency to copy other people's actions. Although automatic imitation builds rapport and affiliation between individuals, we do not copy actions indiscriminately. Instead, copying behaviors are guided by a selection mechanism, which inhibits some actions and prioritizes others. To date, the neural underpinnings of the inhibition of automatic imitation and differences between the sexes in imitation control are not well understood. Previous studies involved small sample sizes and low statistical power, which produced mixed findings regarding the involvement of domain-general and domain-specific neural architectures. Here, we used data from Experiment 1 ( N = 28) to perform a power analysis to determine the sample size required for Experiment 2 ( N = 50; 80% power). Using independent functional localizers and an analysis pipeline that bolsters sensitivity, during imitation control we show clear engagement of the multiple-demand network (domain-general), but no sensitivity in the theory-of-mind network (domain-specific). Weaker effects were observed with regard to sex differences, suggesting that there are more similarities than differences between the sexes in terms of the neural systems engaged during imitation control. In summary, neurocognitive models of imitation require revision to reflect that the inhibition of imitation relies to a greater extent on a domain-general selection system rather than a domain-specific system that supports social cognition.


Genomics of Methylotrophy in Gram-Positive Methylamine-Utilizing Bacteria.
Gram-positive methylotrophic bacteria have been known for a long period of time, some serving as model organisms for characterizing the specific details of methylotrophy pathways/enzymes within this group. However, genome-based knowledge of methylotrophy within this group has been so far limited to a single species, Bacillus methanolicus (Firmicutes). The paucity of whole-genome data for Gram-positive methylotrophs limits our global understanding of methylotrophy within this group, including their roles in specific biogeochemical cycles, as well as their biotechnological potential. Here, we describe the isolation of seven novel strains of Gram-positive methylotrophs that include two strains of Bacillus and five representatives of Actinobacteria classified within two genera, Arthrobacter and Mycobacterium. We report whole-genome sequences for these isolates and present comparative analysis of the methylotrophy functional modules within these genomes. The genomic sequences of these seven novel organisms, all capable of growth on methylated amines, present an important reference dataset for understanding the genomic basis of methylotrophy in Gram-positive methylotrophic bacteria. This study is a major contribution to the field of methylotrophy, aimed at closing the gap in the genomic knowledge of methylotrophy within this diverse group of bacteria.


Sphingolipid rheostat alterations related to transformation can be exploited for specific induction of lysosomal cell death in murine and human glioma.
The search for cancer cell-specific targets suffers from a lack of integrative approaches that take into account the relative contributions of several mechanisms or pathways involved in cell death. A systematic experimental and computational comparison of murine glioma cells with astrocytes, their nontransformed counterparts, identified differences in the sphingolipid (SL) rheostat linked to an increased lysosomal instability in glioma cells. In vitro and in silico analyses indicate that sphingosine metabolized in lysosomes was preferentially recycled into ceramide, the prodeath member of the rheostat, in astrocytes. In glioma cells, it preferentially was used for production of the prosurvival sphingosine-1-phosphate (S1P). A combination of tumor necrosis factor alpha (TNF-alpha), lipopolysaccharide (LPS), and interferon gamma (IFN-gamma) strongly decreased S1P production that resulted in abnormal lysosome enlargement and cell death associated with mitochondrial dysfunction of glioma cells only. Lack of intracellular S1P in glioma cells was concomitant with protein and lipid accumulation in enlarged lysosomes, indicating a blockade in lysosome recycling, and hence a role for S1P in membrane trafficking. A pharmacological sphingosine kinase inhibitor efficiently replaced the TNF-alpha, LPS, and IFN-gamma combination and killed murine and human glioma cells without affecting astrocytes. Our study provides evidence for a novel mechanism of lysosomal death dependent upon the SL rheostat that can be specifically triggered in glioma cells. It further strengthens the potential of cancer therapies based on specific ceramide pathway alterations.


Prevention of recurrent bacterial cystitis by intravesical administration of hyaluronic acid: a pilot study.
To assess the effect of bladder instillations of hyaluronic acid (HA) on the rate of recurrence of urinary tract infection (UTI). Forty women (mean age 35 years) with a history of recurrent UTI received intravesical instillations of HA (40 mg in 50 mL phosphate-buffered saline) once weekly for 4 weeks then once monthly for 4 months. The UTI status was assessed over a prospective follow-up of 12.4 months and compared with the rates of UTI before instillation, determined by a retrospective review of patient charts covering 15.8 months. After HA treatment no patients had a UTI during the 5-month treatment phase and 28 (70%) were recurrence-free at the end of the follow-up. The mean (sd) rate of UTI per patient-year was 4.3 (1.55) before treatment and 0.3 (0.55) afterward (P < 0.001). The median time to recurrence after HA treatment was 498 days, compared with 96 days beforehand (P < 0.001). The tolerability was excellent, as side-effects were limited to nine patients who reported mild bladder irritation; no patient interrupted the treatment. In this preliminary study, bladder instillations of HA had a significant effect on the rate of UTI in women with a history of recurrent UTIs. The bladder instillation of HA is an acceptable and promising therapeutic alternative in patients with recurrent UTI. Expanded placebo controlled clinical trials examining this application of HA are currently underway.


The role of promastigote secretory gel in the origin and transmission of the infective stage of Leishmania mexicana by the sandfly Lutzomyia longipalpis.
Transmission of leishmaniasis is effected by a specific developmental stage, the metacyclic promastigote. The precursors of metacyclic promastigotes were a distinct subpopulation of parasites, identified for the first time as a new stage in the life-cycle and named leptomonad promastigotes. Microdissection of infected sandflies into 4 midgut regions and foregut allowed precursor-product relationships to be established for amastigote-procyclic promastigote, procyclic-nectomonad promastigote, nectomonad-leptomonad promastigote and leptomonad-metacyclic promastigote developmental switches. Metacyclic promastigotes occurred mainly in the thoracic midgut and cardia, coincident with the accumulation of a promastigote secretory gel (PSG) plug in these anterior regions. The gel-like plug was isolated from flies with mature infections and found to contain predominantly leptomonad promastigotes. The PSG plug also contained the majority (75%) of the total metacyclic promastigote population in the sandflies, which were concentrated at the anterior pole. The PSG plug was found to be the main site of metacyclogenesis, and acted as a reservoir of leptomonad promastigotes from which metacyclic forms differentiated and migrated forward to promote the infective potential of the fly. The PSG plug occluded and distorted the midgut, forcing the stomodeal valve open and affecting the feeding success of the sandflies, such that they experienced difficulty in taking a full meal. Collectively, these data support the role of the PSG in the transmission of leishmaniasis, by conditioning the midgut environment for metacyclogenesis and altering the feeding ability of infected sandflies.


Psychosocial Stress Over the Lifespan, Psychological Factors, and Cardiometabolic Risk in the Community.
The complex relationship between psychosocial stress over the lifetime, psychological factors, and cardiometabolic risk is still poorly understood. Accordingly, our aims were (1) to independently assess the associations between childhood adversity, life-event stress in remote (earlier than the last 5 years), and recent adulthood and cardiometabolic risk, and (2) to determine the role of psychological factors including personality, coping, and depression in these associations. The sample included 2674 adults, aged 35 to 66 years, randomly selected from urban area. Participants underwent a physical examination including the assessment of obesity markers, blood pressure, and blood lipid and glucose levels. Stress during adulthood was determined using the severity scores of 52 stressful life events. Information on adverse childhood experiences and major depressive disorders was collected using semistructured interviews, whereas personality traits and coping mechanisms were evaluated through questionnaires. Both childhood adversity and stress in remote adulthood were associated with elevated body mass index (β [95% confidence interval {CI}] = 0.249 [0.029 to 0.468]; 0.020 [0.006 to 0.034]), waist circumference (β [95% CI] = 0.061 [0.024 to 0.099]; 0.08 [0.04 to 0.11]), and the global cardiometabolic risk score (β [95% CI] = 0.278 [0.017 to 0.540]; 0.017 [0.001 to 0.033]) after adjustment for sociodemographic, lifestyle, and psychological factors. In addition, childhood adversity was associated with low high density lipoprotein levels (β [95% CI] = -0.021 [-0.042 to 0.000]), as well as increased fat mass and systolic blood pressure levels (β [95% CI] = 0.506 [0.165 to 0.846]; 0.952 [0.165 to 1.740]) and stress in remote adulthood with apolipoprotein B levels (β [95% CI] = 0.607 [0.312 to 0.901]). Psychological factors did not account for these associations and were not effect modifiers. Our data demonstrate that psychosocial stress during childhood and remote adulthood favor adiposity and abnormal lipid metabolism.


Intergenerational social mobility: the United States in comparative perspective.
Emily Beller and Michael Hout examine trends in U.S. social mobility, especially as it relates to the degree to which a person's income or occupation depends on his or her parents' background and to the independent contribution of economic growth. They also compare U.S. social mobility with that in other countries. They conclude that slower economic growth since 1975 and the concentration of that growth among the wealthy have slowed the pace of U.S. social mobility. In measuring mobility, economists tend to look at income and sociologists, occupation. The consensus among those measuring occupational mobility is that the average correlation between the occupations of fathers and sons today ranges from 0.30 to 0.40, meaning that most variation in the ranking of occupations is independent of social origins. Those measuring income mobility tend to agree that the elasticity between fathers' and sons' earnings in the United States today is about 0.4, meaning that 40 percent of the difference in incomes between families in the parents' generation also shows up in differences in incomes in the sons' generation. Beller and Hout show that occupational mobility increased during the 1970s, compared with the 1940s-1960s, but there is some evidence to suggest that by the 1980s and 1990s it had declined to past levels. Existing data on income mobility show no clear trends over time, but increases in economic inequality during the 1980s made mobility more consequential by making economic differences between families persist for a longer time. In international comparisons, the United States occupies a middle ground in occupational mobility but ranks lower in income mobility. Researchers have used the variation in mobility to study whether aspects of a country's policy regime, such as the educational or social welfare systems, might be driving these results. There is as yet, however, no scholarly consensus about the sources of cross-national differences in mobility.


VGF-derived peptide, TLQP-21, regulates food intake and body weight in Siberian hamsters.
The Siberian hamster survives winter by decreasing food intake and catabolizing abdominal fat reserves, resulting in a sustained, profound loss of body weight. VGF gene expression is photoperiodically regulated in the hypothalamus with significantly higher expression in lean Siberian hamsters. The aim of this study was to investigate the role of VGF in regulating these seasonal cycles by determining the effects of a VGF-derived peptide (TLQP-21) on food intake and body weight. Acute intracerebroventricular administration of TLQP-21 decreased food intake, and chronic treatment caused a sustained reduction in food intake and body weight and decreased abdominal fat depots. Behavioral analysis revealed that TLQP-21 reduced meal size but not the frequency of feeding bouts, suggesting a primary action on satiety. Hamsters treated with TLQP-21 lost a similar amount of weight as a pair-fed group in which food intake was matched to that of the TLQP-21-treated group. Central or peripheral treatment with TLQP-21 did not produce a significant effect on resting metabolic rate. We conclude that the primary action of TLQP-21 is to decrease food intake rather than increase energy expenditure. TLQP-21 treatment caused a decrease in UCP-1 mRNA in brown adipose tissue, but hypothalamic expression of orexigenic and anorexigenic neuropeptide genes remained unchanged after TLQP-21 treatment, although compensatory increases in NPY and AgRP mRNA were observed in the pair-fed hamsters. The effects of TLQP-21 administration are similar to those in hamsters in short days, suggesting that increased VGF activity may contribute to the hypophagia that underlies the seasonal catabolic state.


Student-teacher relationship quality moderates longitudinal associations between child temperament and behavior problems.
Relatively few studies have focused on relational factors within the school environment that could moderate the associations between temperament and behavior problems. The purpose of this longitudinal study was to examine whether student-teacher relationship quality (i.e., Closeness, Conflict, and Dependency) moderated longitudinal associations between child temperament (i.e., Negative Affectivity, Surgency/Extraversion, and Effortful Control) and externalizing and internalizing problems. The participants were 744 elementary school students (mean age 8.4 years at study entry) followed over 3 years (four measurement times). Parents assessed their child's temperament at study entry (T1) and the children reported their self-perceptions of problems 3 years later (T4). Student-teacher relationship quality was constructed as a latent variable based on ratings from three different teachers over 3 years (T1, T2, T3). Latent moderated structural equations revealed that a low level of Surgency/Extraversion was associated with less internalizing problems but only for children experiencing a high level of Closeness with their teachers. A high level of Negative Affectivity was associated with more internalizing problems, but only for children experiencing a high level of Conflict or a high level of Dependency with their teachers. Student-teacher relationship quality did not moderate the associations between temperament and externalizing problems. Improving student-teacher relationship quality through focused interventions thus represents a useful strategy.


Genetic methods for illuminating the function of neural circuits.
Guided by the notion that biology itself offers some of the most incisive tools for studying biological systems, neurophysiologists rely increasingly on cell biological mechanisms and materials encoded in DNA to visualize and control the activity of neurons in functional circuits. Optical reporter proteins can broadcast the operational states of genetically designated cells and synapses; remote-controlled effectors can suppress or induce electrical activity. Many challenges, however, remain. These include the development of novel gene expression systems that target reporters and effectors to functionally relevant neuronal ensembles, the capacity to monitor and manipulate multiple populations of neurons in parallel, the ability to observe and elicit precisely timed action potentials, and the power to communicate with genetically designated target neurons through electromagnetic signals other than light.


A new polysaccharidic gel matrix for drug delivery: preparation and mechanical properties.
A new type of hydrogel was prepared, under controlled conditions, by diffusion of Ca(II) ions into a solution of the carboxylated derivative of Scleroglucan (Sclerox). The obtained hydrogel was loaded with Theophylline (TPH) and Myoglobin (MGB), two model drugs of remarkably different steric hindrance, and also used, after freeze drying, for the preparation of tablets. Release studies were carried out on both the freshly prepared gel and on the tablets. As far as the gel systems experiments are concerned, the delivery profiles resulted to be deeply dependent on the molecular dimensions of the loaded molecules; TPH was easily released while the larger tested molecule (MGB) remained partially entrapped within the three-dimensional network. Furthermore, in the case of MGB, the release was dependent also on polymer concentration (c(p)): at the highest investigated c(p) value a corresponding lowest delivery of the guest molecule was observed. This effect of polymer concentration on the rate of delivery was studied applying three different mathematical approaches: the one that better fitted the experimental release profile allowed to support the explanation of the mechanism involved in the observed two-step delivery that has been related to the drug trapping inside the clusters of the gel network. The delivery profiles from the tablets showed how the release, in this case, could be related, essentially, to the molecular dimensions of the guest molecules, independently on the c(p) used to prepare the starting hydrogel. TPH was completely delivered in a few hours while the MGB was almost unable to diffuse out of the matrix and more than 80% resulted entrapped in the network for at least 24 h. The novel hydrogel, at different c(p), was also characterized by means of a texture analyzer to inspect its mechanical properties. According to the compression data, the hardness, the work of cohesion and the work of adhesion of the networks were estimated. Furthermore, by means of relaxation experiments, analysed applying the generalized Maxwell model, the gels can be classified as solid viscoelastic materials and the mechanical spectra indicated a predominance of the viscous behaviour, while the Young modulus, E0, as expected, was found to increase with polymer concentration.


Whole genome amplification and exome sequencing of archived schistosome miracidia.
Adult schistosomes live in the blood vessels and cannot easily be sampled from humans, so archived miracidia larvae hatched from eggs expelled in feces or urine are commonly used for population genetic studies. Large collections of archived miracidia on FTA cards are now available through the Schistosomiasis Collection at the Natural History Museum (SCAN). Here we describe protocols for whole genome amplification of Schistosoma mansoni and Schistosome haematobium miracidia from these cards, as well as real time PCR quantification of amplified schistosome DNA. We used microgram quantities of DNA obtained for exome capture and sequencing of single miracidia, generating dense polymorphism data across the exome. These methods will facilitate the transition from population genetics, using limited numbers of markers to population genomics using genome-wide marker information, maximising the value of collections such as SCAN.


Regional differential effects of chronic nicotine on brain alpha 4-containing and alpha 6-containing receptors.
Chronic nicotine upregulates central nicotinic acetylcholine receptors (nAChRs), a plasticity process thought to contribute to its addictive properties. To analyze this process in vivo, we chronically exposed mice to nicotine using minipump delivering nicotine at concentrations close to those found in tobacco smokers. Binding studies show upregulation of high-affinity nAChRs after 21 days of treatment in cortical areas, caudate putamen, nucleus accumbens, hippocampus, ventral tegmental area, and superior colliculi. No upregulation was observed in thalamus and discrete cortical areas. Using wild type and alpha 6-/- mice, we observed a downregulation of alpha 6*-nAChRs in superior colliculi and no effects in other structures. The complex pattern of upregulation/downregulation observed in this study depends on both nAChR composition and regional distribution.


Exocytosis precedes and predicts the increase in growth in oscillating pollen tubes.
We examined exocytosis during oscillatory growth in lily (Lilium formosanum and Lilium longiflorum) and tobacco (Nicotiana tabacum) pollen tubes using three markers: (1) changes in cell wall thickness by Nomarski differential interference contrast (DIC), (2) changes in apical cell wall fluorescence in cells stained with propidium iodide (PI), and (3) changes in apical wall fluorescence in cells expressing tobacco pectin methyl esterase fused to green fluorescent protein (PME-GFP). Using PI fluorescence, we quantified oscillatory changes in the amount of wall material from both lily and tobacco pollen tubes. Measurement of wall thickness by DIC was only possible with lily due to limitations of microscope resolution. PME-GFP, a direct marker for exocytosis, only provides information in tobacco because its expression in lily causes growth inhibition and cell death. We show that exocytosis in pollen tubes oscillates and leads the increase in growth rate; the mean phase difference between exocytosis and growth is -98 degrees +/- 3 degrees in lily and -124 degrees +/- 4 degrees in tobacco. Statistical analyses reveal that the anticipatory increase in wall material predicts, to a high degree, the rate and extent of the subsequent growth surge. Exocytosis emerges as a prime candidate for the initiation and regulation of oscillatory pollen tube growth.


Outcome in 104 pubovaginal slings using freeze-dried allograft fascia lata from a single tissue bank.
We describe our experience with the use of allograft fascia lata for the treatment of stress urinary incontinence. One hundred and four patients underwent allograft fascia lata pubovaginal slings. Preoperatively, all were evaluated by a detailed urogynecologic evaluation, voiding diary, and pelvic examination. The pubovaginal sling was performed using a 2x15-cm freeze-dried nonirradiated cadaveric fascia lata specimen. Outcome measures were assessed by a urogynecologic questionnaire, pad usage, and disease-specific quality-of-life questionnaires. Eighty-eight percent (91 of 104) responded to a mailed urogynecology and disease-specific quality-of-life questionnaire with an average follow-up period of 19. 4 +/- 10.3 months. The mean preoperative daily pad usage was 4.6 +/- 3.0, postoperatively pad usage was 1.1 +/- 1.4 (P < 0.0001). Urge incontinence resolved in 41% (n = 24) of the 59 patients who complained of this preoperatively. Eighty-seven percent of the responders indicated that urinary incontinence was not substantially affecting their daily life. As in our preliminary report, the use of freeze-dried allograft pubovaginal sling continues to provide good results without adverse outcomes. A prospective, randomized comparison of autologous versus allograft slings and a review of preparation techniques used by tissue banks are needed.


Comprehensive molecular phylogeny of the grassbirds and allies (Locustellidae) reveals extensive non-monophyly of traditional genera, and a proposal for a new classification.
The widespread Old World avian family Locustellidae ('grassbirds and allies') comprises 62 extant species in 11 genera. In the present study, we used one mitochondrial and, for most species, four nuclear loci to infer the phylogeny of this family. We analysed 59 species, including the five previously unsampled genera plus two genera that had not before been analysed in a densely sampled dataset. This study revealed extensive disagreement with current taxonomy; the genera Bradypterus, Locustella, Megalurus, Megalurulus and Schoenicola were all found to be non-monophyletic. Non-monophyly was particularly pronounced for Megalurus, which was widely scattered across the tree. Three of the five monotypic genera (Amphilais, Buettikoferella and Malia) were nested within other genera; one monotypic genus (Chaetornis) formed a clade with one of the two species of Schoenicola; whereas the position of the fifth monotypic genus (Elaphrornis) was unresolved. Robsonius was confirmed as sister to the other genera. We propose a phylogenetically informed revision of genus-level taxonomy, including one new generic name. Finally, we highlight several non-monophyletic species complexes and deep intra-species divergences that point to conflict in taxonomy and suggest an underestimation of current species diversity in this group.


Secondary Bilateral Angle Closure Glaucoma due to Topiramate.
We examined a 39-year-old female with severe headache and blurred vision. She was on topiramate, 50 mg once a day for one week because of migraine. Periorbital edema, chemosis, myopia, high intraocular pressures, and shallow anterior chambers were present at the initial examination. Iridocorneal angles were closed, ultrasound showed choroidal effusions. We stopped topiramate and started antiglaucoma treatment. After one week the intraocular pressure was 10 mm Hg in both eyes without treatment. A new ultrasound showed no choroidal effusions. Topiramate has been associated with acute secondary angle closure glaucoma as an idiosyncratic reaction to the drug. Physicians prescribing topiramate need to alert patients of this potential sight-threatening idiosyncratic reaction.


Trends in Medical Aid in Dying in Oregon and Washington.
The combined 28 years of data of medical aid in dying (MAID) between Oregon (OR) and Washington (WA) are the most comprehensive in North America. No reports to date have compared MAID use in different US states. To evaluate and compare patterns of MAID use between the states with the longest-running US death with dignity programs. A retrospective observational cohort study of OR and WA patients with terminal illness who received prescriptions as part of their states' legislation allowing MAID. All published annual reports, from 1998 to 2017 in OR and from 2009 to 2017 in WA, were reviewed. A total of 3368 prescriptions were included. Number of deaths from self-administration of lethal medication vs number of prescriptions written. A combined 3368 prescriptions were written in OR and WA, with 2558 patient deaths from lethal ingestion (76.0%). Of the 2558 patients, most were male (1311 [51.3%]), older than 65 years (1851 [72.4%]), and non-Hispanic white (2426 [94.8%]). The most common underlying illnesses were cancer (1955 [76.4%]), neurologic illness (261 [10.2%]), lung disease (144 [5.6%]), and heart disease (117 [4.6%]). Loss of autonomy (2235 [87.4%]), impaired quality of life (2203 [86.1%]), and loss of dignity (1755 [68.6%]) were the most common reasons for pursuing MAID. Time between drug intake to coma ranged from 1 to 660 minutes and time from drug intake to death ranged from 1 to 6240 minutes. In the 1557 patients for whom rates of complications were reported, 1494 (96.0%) did not experience a complication (592 of 626 [94.6%] in OR and 902 of 931 [96.8%] in WA). Eight patients (<0.5%) regained consciousness after drug ingestion in OR. Annual rates per year for percentage of patients who received a prescription ingesting the prescribed medication ranged from 48% to 87%, with no significant time trend in OR (adjusted odds ratio per year, 1.01; 95% CI, 0.99-1.02; P = .59) but with an increase over time in WA (adjusted odds ratio per year, 1.13; 95% CI, 1.08-1.19; P < .001). In both OR and WA there were increases in the number of patient deaths due to MAID per 1000 deaths over time. In this study, MAID results in Oregon and Washington were similar, although MAID use measured as a percentage of patients prescribed lethal medications and then self-administering them increased only in WA. Most patients who acquired lethal prescriptions had cancer or terminal illnesses that are difficult to palliate and lead to loss of autonomy, dignity, and quality of life.


Pro-Apoptotic Activity of French Polynesian Padina pavonica Extract on Human Osteosarcoma Cells.
Recently, seaweeds and their extracts have attracted great interest in the pharmaceutical industry as a source of bioactive compounds. Studies have demonstrated the cytotoxic activity of macroalgae towards different types of cancer cell models, and their consumption has been suggested as a chemo-preventive agent against several cancers such as breast, cervix and colon cancers. Reports relevant to the chemical properties of brown algae Padina sp. are limited and those accompanied to a comprehensive evaluation of the biological activity on osteosarcoma (OS) are non existent. In this report, we explored the chemical composition of French Polynesian Padina pavonica extract (EPP) by spectrophotometric assays (total phenolic, flavonoid and tannin content, and antioxidant activity) and by gas chromatography-mass spectrometry (GC-MS) analysis, and provided EPP lipid and sterols profiles. Several compounds with relevant biological activity were also identified that suggest interesting pharmacological and health-protecting effects for EPP. Moreover, we demonstrated that EPP presents good anti-proliferative and pro-apoptotic activities against two OS cell lines, SaOS-2 and MNNG, with different cancer-related phenotypes. Finally, our data suggest that EPP might target different properties associated with cancer development and aggressiveness.


Relative performance of antimicrobial susceptibility assays on clinical Escherichia coli isolates from animals.
The assessment of antimicrobial resistance in bacteria derived from animals is often performed using the disc diffusion assay. However broth-microdilution is the preferred assay for national antimicrobial resistance surveillance programs. This study aimed to evaluate the accuracy of disc diffusion relative to broth-microdilution across a panel of 12 antimicrobials using data from a collection of 994 clinical Escherichia coli isolates from animals. Disc diffusion performance was evaluated by diagnostic sensitivity, specificity, likelihood ratio pairs and receive-operating characteristic (ROC) analysis. Data was dichotomised using CLSI susceptible and resistant clinical breakpoints. In addition, disc diffusion breakpoints produced using diffusion Breakpoint Estimation Testing Software (dBETS) were evaluated. Analysis revealed considerable variability in performance estimates for disc diffusion susceptible and resistant breakpoints (AUC ranges: 0.78-0.99 and 0.92-1.0, respectively) across the panel of antimicrobials. Ciprofloxacin, tetracycline, and ampicillin estimates were robust across both breakpoints, whereas estimates for several antimicrobials including amoxicillin-clavulanic acid, cefoxitin and gentamicin were less favourable using susceptible breakpoints. Overall performance estimates were moderately improved when dBETS susceptible breakpoints were applied. For most antimicrobials, disc diffusion was accurate at predicting resistance of clinical E. coli from animals that could otherwise be determined by broth-microdilution. While disc diffusion is suboptimal for assessing the proportion of fully susceptible isolates for some drugs, sensitivity and specificity estimates provided here allow for the use of standard formula to correct this. For this reason, disc diffusion has applicability in national surveillance provided the performance of the assay is taken into account.


Genetic linkage of systemic lupus erythematosus to 13q32 in African American families with affected male members.
Systemic lupus erythematosus (SLE) is a complex autoimmune disorder involving genetic and environmental factors. Previously, our group showed that SLE females with affected male relatives have higher prevalence of renal disease than SLE females with no affected male relatives in a sample of 372 individuals from 159 families. By adding 392 individuals from 181 new families, we replicated this finding in the largest collection of families with affected males, confirming our hypothesis that multiplex SLE families with at least one affected male member ("male families") comprise a distinct subpopulation of SLE multiplex families. We studied 64 male families by a genome-wide scan for SLE and found the largest signal (lod=3.08) at 13q32 in 18 African American male families using an affected-relative-pair model-free linkage method. Closer examination of IBD sharing at this region suggested a dominant mode of inheritance. Multipoint model-based linkage analysis generated a lod score of 3.13 in the same chromosomal region with a low-disease allele frequency of 0.0004 and a disease penetrance of 0.5 for the 18 African American male families. We performed fine mapping in these and three additional African American male families and the SLE predisposing locus was localized to a region tightly linked to the marker D13S892. We have therefore confirmed the linkage of SLE to 13q32, which was reported previously, and suggested that an SLE susceptibility gene in this region is specific to predisposition of African Americans to a specific form of SLE, with males at high risk.


Genetic variations in the TERT and CLPTM1L gene region and gastrointestinal stromal tumors risk.
Recent studies have suggested polymorphisms in the TERT and CLPTM1L region are associated with carcinogenesis of many distinct cancer types, including gastrointestinal cancers. However, the contribution of polymorphisms in the TERT and CLPTM1L gene region to gastrointestinal stromal tumors (GISTs) risk is still unknown. We tested the six tagSNPs on TERT and CLPTM1L region with GIST risk, using a population-based, two-stage, case-control study in 2,000 subjects. Functional validation was conducted to validate our findings of TERT rs2736098 and explore its influence on relative telomere length (RTL) in GIST cells. It showed that variant rs2736098 was significantly associated with increased risk of GIST (per allele OR = 1.29, 95% CI: 1.14-1.47, P = 7.03 × 10-5). The difference remain significant after Bonferroni correction (P = 7.03 × 10-5 * 6 = 4.2 × 10-4). Real-time PCR showed carriers of genotype CC have the longest RTL, following by carriers of genotype CT, while carriers of genotype TT have the shortest RTL in GIST tissues (P < 0.001). Our data provide evidence to implicate TERT rs2736098 polymorphism as a novel susceptibility factor for GIST risk.


Dynamic chromatin organization during foregut development mediated by the organ selector gene PHA-4/FoxA.
Central regulators of cell fate, or selector genes, establish the identity of cells by direct regulation of large cohorts of genes. In Caenorhabditis elegans, foregut (or pharynx) identity relies on the FoxA transcription factor PHA-4, which activates different sets of target genes at various times and in diverse cellular environments. An outstanding question is how PHA-4 distinguishes between target genes for appropriate transcriptional control. We have used the Nuclear Spot Assay and GFP reporters to examine PHA-4 interactions with target promoters in living embryos and with single cell resolution. While PHA-4 was found throughout the digestive tract, binding and activation of pharyngeally expressed promoters was restricted to a subset of pharyngeal cells and excluded from the intestine. An RNAi screen of candidate nuclear factors identified emerin (emr-1) as a negative regulator of PHA-4 binding within the pharynx, but emr-1 did not modulate PHA-4 binding in the intestine. Upon promoter association, PHA-4 induced large-scale chromatin de-compaction, which, we hypothesize, may facilitate promoter access and productive transcription. Our results reveal two tiers of PHA-4 regulation. PHA-4 binding is prohibited in intestinal cells, preventing target gene expression in that organ. PHA-4 binding within the pharynx is limited by the nuclear lamina component EMR-1/emerin. The data suggest that association of PHA-4 with its targets is a regulated step that contributes to promoter selectivity during organ formation. We speculate that global re-organization of chromatin architecture upon PHA-4 binding promotes competence of pharyngeal gene transcription and, by extension, foregut development.


Uterine and vaginal organ growth requires epidermal growth factor receptor signaling from stroma.
Estrogens are crucial for growth and function of the female genital tract. Recently, we showed that induction of uterine epithelial proliferation by estradiol is a paracrine event requiring an estrogen receptor-positive stroma. Growth factors [such as EGF (epidermal growth factor) ligands] are likely paracrine mediators, which may directly or indirectly regulate epithelial proliferation in estrogen target organs via cell-cell interactions. In this report, we used mice with a null mutation in their EGF receptor (EGFR) to examine the role of EGFR signaling in growth of the uterus and vagina and in estrogen-induced uterine and vaginal epithelial proliferation. When WT and EGFR-knockout (EGFR-KO) uteri and vaginae were grown as renal capsule grafts in nude mice, growth of uterine and vaginal grafts of EGFR-KO mice was reduced, compared with their WT counterparts. Grafts of both EGFR-KO uteri and vaginae were about one third smaller (wet weight) than their corresponding WT organs, even though differentiation of both epithelium and mesenchyme were normal in both cases. Both wild-type and EGFR-KO vaginal grafts contained within their lumina alternating layers of cornified and mucified epithelial cell layers, indicating cyclic alteration of epithelial differentiation. In response to estradiol treatment, stromal cell labeling index (LI), as assessed by incorporation of 3H-thymidine, was severely depressed in EGFR-KO uterine and vaginal grafts vs. stromal cell LI in WT uterine and vaginal grafts. Unexpectedly, epithelium of both EGFR-KO and wild-type grafts responded comparably to estradiol with a marked elevation (approximately 7-fold overall) of epithelial LI in response to estradiol in uterine and vaginal epithelia. These data supported the hypothesis that overall uterine and vaginal organ growth, in response to estrogen, required EGFR signaling for DNA synthesis in the fibromuscular stroma, whereas EGFR signaling was not essential for estrogen-induced epithelial growth in the uterus and vagina.


Best (but oft-forgotten) practices: identifying and accounting for regression to the mean in nutrition and obesity research.
Regression to the mean (RTM) is a statistical phenomenon where initial measurements of a variable in a nonrandom sample at the extreme ends of a distribution tend to be closer to the mean upon a second measurement. Unfortunately, failing to account for the effects of RTM can lead to incorrect conclusions on the observed mean difference between the 2 repeated measurements in a nonrandom sample that is preferentially selected for deviating from the population mean of the measured variable in a particular direction. Study designs that are susceptible to misattributing RTM as intervention effects have been prevalent in nutrition and obesity research. This field often conducts secondary analyses of existing intervention data or evaluates intervention effects in those most at risk (i.e., those with observations at the extreme ends of a distribution). To provide best practices to avoid unsubstantiated conclusions as a result of ignoring RTM in nutrition and obesity research. We outlined best practices for identifying whether RTM is likely to be leading to biased inferences, using a flowchart that is available as a web-based app at https://dustyturner.shinyapps.io/DecisionTreeMeanRegression/. We also provided multiple methods to quantify the degree of RTM. Investigators can adjust analyses to include the RTM effect, thereby plausibly removing its biasing influence on estimating the true intervention effect. The identification of RTM and implementation of proper statistical practices will help advance the field by improving scientific rigor and the accuracy of conclusions. This trial was registered at clinicaltrials.gov as NCT00427193.


Preservation of the Epiphysis and Growth Plate in the Surgical Management of Femoral Osteosarcoma in a Skeletally Immature Patient by Intercalary Resection and Biological Reconstruction: A Case Report.
Osteosarcoma arises most frequently in the metaphysis around the knee and its management by limb salvage surgery in skeletally immature pediatric patients is extremely challenging. Common reconstructive methods such as endoprosthetic or biological reconstruction are not fully capable of dealing with durability-related and growth-related problems and their functional outcomes are not as good as those seen in adult cases. A definitive limb salvaging procedure in children that outperforms amputation or rotationplasty has not yet been established. Herein, we report a case of stage IV osteosarcoma in the femur of a 7-year-old boy that was safely managed with intercalary resection preserving the distal femoral growth plate and epiphysis, followed by biological reconstruction using a frozen tumor-devitalized autograft. Good response to preoperative chemotherapy and the diaphyseal location of the tumor enabled us to perform a tumor resection that spared the growth plate and preserved the native knee joint structure. Plate fixation over the growth plate was terminated by removing the locking screws in the epiphysis after 44 months, which restored growth capacity to some extent. At 50 months postoperatively, no recurrence or progression of the disease was observed. The patient uses an extension shoe and reports having little discomfort in his daily life despite having a restricted range of motion and limb length discrepancy. In conclusion, limb salvage with biological reconstruction in skeletally immature patients can provide an acceptable functional outcome, including minimized limb length discrepancy, if critical damage to the growth plate and articular components can be avoided.


A comparison of pain experienced by patients treated with labial and lingual orthodontic appliances.
The aim of this prospective longitudinal study was to compare pain experiences among Chinese adult patients treated with labial and lingual orthodontic appliances. Sixty patients, 30 with labial appliances (18 females and 12 males, mean age 20.33 years, SD +/- 4.205) and 30 with lingual appliances (22 females and 8 males, mean age 21.63 years, SD +/- 2.236), rated their overall pain experience on a 100 mm visual analogue scale (VAS) at three time points: 1 week (T(1)), 1 month (T(2)), and 3 months (T(3)) after bracket placement. In addition, on a separate 100 mm VAS, they rated their pain experience at the locations of the tongue, lips, cheeks, gums, face, and jaw at T(1), T(2), and T(3). Changes in pain VAS were conducted using Friedman analysis of variance, area under the curve (AUC) analysis and the data were compared using a t-test. There was no significant difference in global ratings of pain among those treated with labial or lingual appliances (P > 0.05). Among both groups, global ratings of pain decreased over the study period (P < 0.001). Patients treated with lingual appliances reported higher ratings of tongue pain (P < 0.001), while those treated with labial appliances reported higher ratings of lip (P < 0.001) and cheek (P < 0.001) pain. The findings indicate that patients treated with labial and lingual appliances rate similarly the level of overall pain they experience during treatment. Ratings of overall pain experienced decreased for both treatment groups with time. However, ratings of pain differed at various sites with respect to the type of orthodontic appliance. These findings have implications in informing patients' treatment decision-making processes regarding labial and lingual appliances and in the management of discomfort associated with different treatment modalities.


Expression of two nonallelic type II procollagen genes during Xenopus laevis embryogenesis is characterized by stage-specific production of alternatively spliced transcripts.
The pattern of type II collagen expression during Xenopus laevis embryogenesis has been established after isolating specific cDNA and genomic clones. Evidence is presented suggesting that in X. laevis there are two transcriptionally active copies of the type II procollagen gene. Both genes are activated at the beginning of neurula stage and steady-state mRNA levels progressively increase thereafter. Initially, the transcripts are localized to notochord, somites, and the dorsal region of the lateral plate mesoderm. At later stages of development and parallel to increased mRNA accumulation, collagen expression becomes progressively more confined to chondrogenic regions of the tadpole. During the early period of mRNA accumulation, there is also a transient pattern of expression in localized sites that will later not undergo chondrogenesis, such as the floor plate in the ventral neural tube. At later times and coincident with the appearance of chondrogenic tissues in the developing embryo, expression of the procollagen genes is characterized by the production of an additional, alternatively spliced transcript. The alternatively spliced sequences encode the cysteine-rich globular domain in the NH2-propeptide of the type II procollagen chain. Immunohistochemical analyses with a type II collagen monoclonal antibody documented the deposition of the protein in the extracellular matrix of the developing embryo. Type II collagen expression is therefore temporally regulated by tissue-specific transcription and splicing factors directing the synthesis of distinct molecular forms of the precursor protein in the developing Xenopus embryo.


Mortality After Repeat Revascularization Following PCI or CABG for Left Main Disease: The EXCEL Trial.
The aim of this study was to investigate the incidence and impact on mortality of repeat revascularization after index percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG) for left main coronary artery disease (LMCAD). The impact on mortality of the need of repeat revascularization following PCI or CABG in patients with unprotected LMCAD is unknown. All patients with LMCAD and site-assessed low or intermediate SYNTAX (Synergy Between PCI With Taxus and Cardiac Surgery) scores randomized to PCI (n = 948) or CABG (n = 957) in the EXCEL (Evaluation of XIENCE Versus Coronary Artery Bypass Surgery for Effectiveness of Left Main Revascularization) trial were included. Repeat revascularization events were adjudicated by an independent clinical events committee. The effect of repeat revascularization on mortality through 3-year follow-up was examined in time-varying Cox regression models. During 3-year follow-up, there were 346 repeat revascularization procedures among 185 patients. PCI was associated with higher rates of any repeat revascularization (12.9% vs. 7.6%; hazard ratio: 1.73; 95% confidence interval: 1.28 to 2.33; p = 0.0003). Need for repeat revascularization was independently associated with increased risk for 3-year all-cause mortality (adjusted hazard ratio: 2.05; 95% confidence interval: 1.13 to 3.70; p = 0.02) and cardiovascular mortality (adjusted hazard ratio: 4.22; 95% confidence interval: 2.10 to 8.48; p < 0.0001) consistently after both PCI and CABG (pint = 0.85 for both endpoints). Although target vessel revascularization and target lesion revascularization were both associated with an increased risk for mortality, target vessel non-target lesion revascularization and non-target vessel revascularization were not. In the EXCEL trial, repeat revascularization during follow-up was performed less frequently after CABG than PCI and was associated with increased mortality after both procedures. Reducing the need for repeat revascularization may further improve long-term survival after percutaneous or surgical treatment of LMCAD. (EXCEL Clinical Trial; NCT01205776).


Cost Consequence Analysis of Implementing the Low Risk Ankle Rule in Emergency Departments.
Implementation of the Low Risk Ankle Rule can safely reduce radiographs for children with acute ankle injuries. The main objective of this study is to examine the costs and consequences of implementing the rule. For children aged 3 to 16 years and with an acute ankle injury, we collected data on health care provider visits, imaging, and treatment at the index emergency department (ED) visit and days 7 and 28 post-ED discharge. This was done during 3 consecutive 6-month phases at 6 EDs. After the baseline phase 1, the Low Risk Ankle Rule was introduced in phases 2 and 3 in 3 intervention EDs, but not in the 3 pair-matched control EDs. We compared the effect of the Low Risk Ankle Rule on health care and patient-paid costs, the proportion of radiographs ordered, the proportion of missed clinically important fractures, and the follow-up use of health care resources. We enrolled 2,151 children with ankle injuries, 1,055 at the intervention and 1,096 at the control EDs. Health care costs were $36.93 less per patient at intervention compared with control sites (P=.02). Out-of-pocket costs to the patients were $2.09 more per patient at intervention sites (P=.30). In intervention versus control sites, the main contributor to cost reduction was the 22.9% reduction in ankle radiography. Furthermore, there were no significant differences in the frequency of missed clinically important fractures (0.1% versus 0.9%) or follow-up use of health care resources. Widespread implementation of the Low Risk Ankle Rule may lead to reduction of unnecessary radiographs for children and result in cost savings.


PyClone: statistical inference of clonal population structure in cancer.
We introduce PyClone, a statistical model for inference of clonal population structures in cancers. PyClone is a Bayesian clustering method for grouping sets of deeply sequenced somatic mutations into putative clonal clusters while estimating their cellular prevalences and accounting for allelic imbalances introduced by segmental copy-number changes and normal-cell contamination. Single-cell sequencing validation demonstrates PyClone's accuracy.


The effect of caffeine on tear secretion.
Caffeine, probably the most widely consumed psychoactive substance, is claimed to have conflicting effects on some tear film dynamics. This study sought to investigate the effect of orally ingested caffeine on tear secretion. In an examiner-masked, placebo-controlled, crossover experimental model, the effect of caffeine intake on tear secretion was studied in 41 healthy volunteers aged 20 to 26 years (mean, 23.0 ± 2.1 years). Participants were randomly assigned into two groups, A and B, to receive two different treatments in two sessions. Subjects in group A were exposed to 5.0 mg/kg body weight of caffeine dissolved in 200 mL of water on their first visit, whereas those in group B were exposed to 200 mL of water. On the second visit, however, the order of treatment was reversed. Schirmer 1 scores were measured repeatedly at 45, 90, 135, and 180 minutes after treatment. The baseline Schirmer 1 scores were compared with posttreatment scores. Schirmer 1 scores increased after caffeine intake. The increase was statistically significant at 45 and 90 minutes (p < 0.05) after caffeine intake. Age, body mass, and blood pressure had no correlation with Schirmer 1 scores (Spearman correlation test, p > 0.05). There was no influence of gender in caffeine's effect on tear secretion (F = 0.994, p = 0.399). From our study, orally ingested caffeine appears to stimulate tear secretion in healthy non-dry eye subjects.


Pedicle screw-based posterior dynamic stabilisation of the lumbar spine: in vitro cadaver investigation and a finite element study.
Pedicle screw-based dynamic constructs either benefit from a dynamic (flexible) interconnecting rod or a dynamic (hinged) screw. Both types of systems have been reported in the literature. However, reports where the dynamic system is composed of two dynamic components, i.e. a dynamic (hinged) screw and a dynamic rod, are sparse. In this study, the biomechanical characteristics of a novel pedicle screw-based dynamic stabilisation system were investigated and compared with equivalent rigid and semi-rigid systems using in vitro testing and finite element modelling analysis. All stabilisation systems restored stability after decompression. A significant decrease in the range of motion was observed for the rigid system in all loadings. In the semi-rigid construct the range of motion was significantly less than the intact in extension, lateral bending and axial rotation loadings. There were no significant differences in motion between the intact spine and the spine treated with the dynamic system (P>0.05). The peak stress in screws was decreased when the stabilisation construct was equipped with dynamic rod and/or dynamic screws.


Isotopic evidence of multiple controls on atmospheric oxidants over climate transitions.
The abundance of tropospheric oxidants, such as ozone (O3) and hydroxyl (OH) and peroxy radicals (HO2 + RO2), determines the lifetimes of reduced trace gases such as methane and the production of particulate matter important for climate and human health. The response of tropospheric oxidants to climate change is poorly constrained owing to large uncertainties in the degree to which processes that influence oxidants may change with climate and owing to a lack of palaeo-records with which to constrain levels of atmospheric oxidants during past climate transitions. At present, it is thought that temperature-dependent emissions of tropospheric O3 precursors and water vapour abundance determine the climate response of oxidants, resulting in lower tropospheric O3 in cold climates while HOx (= OH + HO2 + RO2) remains relatively buffered. Here we report observations of oxygen-17 excess of nitrate (a proxy for the relative abundance of atmospheric O3 and HOx) from a Greenland ice core over the most recent glacial-interglacial cycle and for two Dansgaard-Oeschger events. We find that tropospheric oxidants are sensitive to climate change with an increase in the O3/HOx ratio in cold climates, the opposite of current expectations. We hypothesize that the observed increase in O3/HOx in cold climates is driven by enhanced stratosphere-to-troposphere transport of O3, and that reactive halogen chemistry is also enhanced in cold climates. Reactive halogens influence the oxidative capacity of the troposphere directly as oxidants themselves and indirectly via their influence on O3 and HOx. The strength of stratosphere-to-troposphere transport is largely controlled by the Brewer-Dobson circulation, which may be enhanced in colder climates owing to a stronger meridional gradient of sea surface temperatures, with implications for the response of tropospheric oxidants and stratospheric thermal and mass balance. These two processes may represent important, yet relatively unexplored, climate feedback mechanisms during major climate transitions.


Evidence for dissociation of gluconeogenesis stimulated by non-esterified fatty acids and changes in fructose 2,6-bisphosphate in cultured rat hepatocytes.
In order to examine the role of fructose 2,6-bisphosphate (Fru-2,6-P2) in non-esterified-fatty-acid-stimulated gluconeogenesis, Fru-2,6-P2 levels were measured in cultured rat hepatocytes under conditions mimicking the fasted state. After addition of either 1.5 mM-palmitate or 10 nM-glucagon, [U-14C]lactate incorporation into glucose increased 2-fold, but only glucagon suppressed Fru-2,6-P2. Prevention of palmitate oxidation with a carnitine palmitoyltransferase-I inhibitor (2-bromopalmitate) diminished glucose production and Fru-2,6-P2 levels. Addition of exogenous glucose to the media increased Fru-2,6-P2 in a dose-related manner, which was further augmented by addition of palmitate. When Fru-2,6-P2 levels were examined in cells cultured under conditions mimicking the fed state (significantly higher basal Fru-2,6-P2 levels and lower glucose production), palmitate oxidation was associated with a significant fall in Fru-2,6-P2. In conclusion, the present studies have demonstrated a dissociation between fatty-acid-stimulated gluconeogenesis and changes in Fru-2,6-P2 in cultured rat hepatocytes. Further experiments suggest that the accumulation of intracellular hexose 6-phosphate as a result of fatty-acid-stimulated gluconeogenesis masks a putative inhibitory effect of fatty acids on Fru-2,6-P2 concentrations.


Recovery of an environmental Chlamydia strain from activated sludge by co-cultivation with Acanthamoeba sp.
Chlamydiae are a unique group of obligate intracellular bacteria comprising important pathogens of vertebrates as well as symbionts of free-living amoebae. Although there is ample molecular evidence for a huge diversity and wide distribution of chlamydiae in nature, environmental chlamydiae are currently represented by only few isolates. This paper reports the recovery of a novel environmental chlamydia strain from activated sludge by co-cultivation with Acanthamoeba sp. The recovered environmental chlamydia strain UV-7 showed the characteristic morphology of chlamydial developmental stages as revealed by electron microscopy and was identified as a new member of the family Parachlamydiaceae (98.7 % 16S rRNA sequence similarity to Parachlamydia acanthamoebae). Infection studies suggested that Parachlamydia sp. UV-7 is not confined to amoeba hosts but is also able to invade mammalian cells. These findings outline a new straightforward approach to retrieving environmental chlamydiae from nature without prior, tedious isolation and cultivation of their natural host cells, and lend further support to suggested implications of environmental chlamydiae for public health.


A scalable open-source MATLAB toolbox for reconstruction and analysis of multispectral optoacoustic tomography data.
Multispectral photoacoustic tomography enables the resolution of spectral components of a tissue or sample at high spatiotemporal resolution. With the availability of commercial instruments, the acquisition of data using this modality has become consistent and standardized. However, the analysis of such data is often hampered by opaque processing algorithms, which are challenging to verify and validate from a user perspective. Furthermore, such tools are inflexible, often locking users into a restricted set of processing motifs, which may not be able to accommodate the demands of diverse experiments. To address these needs, we have developed a Reconstruction, Analysis, and Filtering Toolbox to support the analysis of photoacoustic imaging data. The toolbox includes several algorithms to improve the overall quantification of photoacoustic imaging, including non-negative constraints and multispectral filters. We demonstrate various use cases, including dynamic imaging challenges and quantification of drug effect, and describe the ability of the toolbox to be parallelized on a high performance computing cluster.


Of plasticity and specificity: dialectics of the micro- and macro-environment and the organ phenotype.
The study of biological form and how it arises is the domain of the developmental biologists; but once the form is achieved, the organ poses a fascinating conundrum for all the life scientists: how are form and function maintained in adult organs throughout most of the life of the organism? That they do appears to contradict the inherently plastic nature of organogenesis during development. How do cells with the same genetic information arrive at, and maintain such different architectures and functions, and how do they keep remembering that they are different from each other? It is now clear that narratives based solely on genes and an irreversible regulatory dynamics cannot answer these questions satisfactorily, and the concept of microenvironmental signaling needs to be added to the equation. During development, cells rearrange and differentiate in response to diffusive morphogens, juxtacrine signals and the extracellular matrix (ECM). These components, which constitute the modular microenvironment, are sensitive to cues from other tissues and organs of the developing embryo as well as from the external macroenvironment. On the other hand, once the organ is formed, these modular constituents integrate and constrain the organ architecture, which ensures structural and functional homeostasis and therefore, organ specificity. We argue here that a corollary of the above is that once the organ architecture is compromised in adults by mutations or by changes in the microenvironment such as aging or inflammation, that organ becomes subjected to the developmental and embryonic circuits in search of a new identity. But since the microenvironment is no longer embryonic, the confusion leads to cancer: hence as we have argued, tumors become new evolutionary organs perhaps in search of an elusive homeostasis.


Redox state and methods to evaluate oxidative stress in liver damage: From bench to bedside.
Oxidative stress is importantly involved in the pathophysiology of various liver diseases. The redox state participates on the course of the inflammatory, metabolic and proliferative liver diseases. The main sources of the reactive oxygen species (ROS) are represented by the mitochondria and cytochrome P450 enzymes in the hepatocyte, Kupffer cells and neutrophils. Cells are provided with efficient molecular strategies to strictly control the intracellular ROS level and to maintain the balance between oxidant and antioxidant molecules. Hepatocyte's proteins, lipids and DNA are among the cellular structures to be affected primarily by ROS and reactive nitrogen species (RNS). This process disrupts at cellular and molecular level the structure-function relationship on liver cells at different sites. Therefore, further studies on the molecular mechanisms of the oxidative stress pathways on liver diseases are urgently required, because they could explain the pathogenesis of various liver disorders. Moreover, new methods to evaluate oxidative stress like the oxidative markers among hepatocytes offers the potential to diagnose the degree of liver injury and ultimately to assess the response to pharmacological therapies. In this review, we discuss the molecular, metabolic and aging aspects of the oxidative stress, and the methods to evaluate oxidative stress on liver damage.


Development of stable monolithic wide-field Michelson interferometers.
Bulk wide-field Michelson interferometers are very useful for high precision applications in remote sensing and astronomy. A stable monolithic Michelson interferometer is a key element in high precision radial velocity (RV) measurements for extrasolar planets searching and studies. Thermal stress analysis shows that matching coefficients of thermal expansion (CTEs) is a critical requirement for ensuring interferometer stability. This requirement leads to a novel design using BK7 and LAK7 materials, such that the monolithic interferometer is free from thermal distortion. The processes of design, fabrication, and testing of interferometers are described in detail. In performance evaluations, the field angle is typically 23.8° and thermal sensitivity is typically -2.6×10(-6)/°C near 550 nm, which corresponds to ∼800 m/s/°C in the RV scale. Low-cost interferometer products have been commissioned in multiple RV instruments, and they are producing high stability performance over long term operations.


Tramadol or morphine administered during operation: a study of immediate postoperative effects after abdominal hysterectomy.
Tramadol may cause awareness and EEG activation during anaesthesia. We compared tramadol with morphine, administered during wound-closure, surmising that tramadol may cause earlier awakening, more rapid recovery, less respiratory depression and equivalent pain relief. Forty patients received nitrous oxide-enflurane for abdominal surgery. At wound closure, patients received tramadol 3 mg kg-1 or morphine 0.2 mg kg-1 and end-tidal enflurane concentrations were maintained at 0.5 kPa until skin closure, whereupon anaesthesia was discontinued. Times to spontaneous respiration, awakening and orientation were similar in the two groups, as were blood-gas tensions, ventilatory frequency, pain scores and incidence of nausea. Half of each group required supplementary analgesia during their 90-min stay in the recovery room. P-deletion counts improved more rapidly in the tramadol group. This study confirms previous reports that tramadol did not antagonize the hypnotic effects of volatile anaesthetics. Tramadol, administered during operation, was as effective as morphine in providing postoperative analgesia while permitting more rapid psychomotor recovery.


A systematic review of positron emission tomography (PET) and positron emission tomography/computed tomography (PET/CT) for the diagnosis of breast cancer recurrence.
Breast cancer (BC) accounts for one-third of all cases of cancer in women in the UK. Current strategies for the detection of BC recurrence include computed tomography (CT), magnetic resonance imaging (MRI) and bone scintigraphy. Positron emission tomography (PET) and, more recently, positron emission tomography/computed tomography (PET/CT) are technologies that have been shown to have increasing relevance in the detection and management of BC recurrence. To review the accuracy of PET and PET/CT for the diagnosis of BC recurrence by assessing their value compared with current practice and compared with each other. MEDLINE and EMBASE were searched from inception to May 2009. Studies were included if investigations used PET or PET/CT to diagnose BC recurrence in patients with a history of BC and if the reference standard used to define the true disease status was histological diagnosis and/or long-term clinical follow-up. Studies were excluded if a non-standard PET or PET/CT technology was used, investigations were conducted for screening or staging of primary breast cancer, there was an inadequate or undefined reference standard, or raw data for calculation of diagnostic accuracy were not available. Quality assessment and data extraction were performed independently by two reviewers. Direct and indirect comparisons were made between PET and PET/CT and between these technologies and methods of conventional imaging, and meta-analyses were carried out. Analysis was conducted separately on patient- and lesion-based data. Subgroup analysis was conducted to investigate variation in the accuracy of PET in certain populations or contexts and sensitivity analysis was conducted to examine the reliability of the primary outcome measures. Of the 28 studies included in the review, 25 presented patient-based data and 7 presented lesion-based data for PET and 5 presented patient-based data and 1 presented patient- and lesion-based data for PET/CT; 16 studies conducted direct comparisons with 12 comparing the accuracy of PET or PET/CT with conventional diagnostic tests and 4 with MRI. For patient-based data (direct comparison) PET had significantly higher sensitivity [89%, 95% confidence interval (CI) 83% to 93% vs 79%, 95% CI 72% to 85%, relative sensitivity 1.12, 95% CI 1.04 to 1.21, p = 0.005] and significantly higher specificity (93%, 95% CI 83% to 97% vs 83%, 95% CI 67% to 92%, relative specificity 1.12, 95% CI 1.01 to 1.24, p = 0.036) compared with conventional imaging tests (CITs)--test performance did not appear to vary according to the type of CIT tested. For patient-based data (direct comparison) PET/CT had significantly higher sensitivity compared with CT (95%, 95% CI 88% to 98% vs 80%, 95% CI 65% to 90%, relative sensitivity 1.19, 95% CI 1.03 to 1.37, p = 0.015), but the increase in specificity was not significant (89%, 95% CI 69% to 97% vs 77%, 95% CI 50% to 92%, relative specificity 1.15, 95% CI 0.95 to 1.41, p = 0.157). For patient-based data (direct comparison) PET/CT had significantly higher sensitivity compared with PET (96%, 95% CI 90% to 98% vs 85%, 95% CI 77% to 91%, relative sensitivity 1.11, 95% CI 1.03 to 1.18, p = 0.006), but the increase in specificity was not significant (89%, 95% CI 74% to 96% vs 82%, 95% CI 64% to 92%, relative specificity 1.08, 95% CI 0.94 to 1.20, p = 0.267). For patient-based data there were no significant differences in the sensitivity or specificity of PET when compared with MRI, and, in the one lesion based study, there was no significant differences in the sensitivity or specificity of PET/CT when compared with MRI. Studies reviewed were generally small and retrospective and this may have limited the generalisability of findings. Subgroup analysis was conducted on the whole set of studies investigating PET and was not restricted to comparative studies. Conventional imaging studies that were not compared with PET or PET/CT were excluded from the review. Available evidence suggests that for the detection of BC recurrence PET, in addition to conventional imaging techniques, may generally offer improved diagnostic accuracy compared with current standard practice. However, uncertainty remains around its use as a replacement for, rather than an add-on to, existing imaging technologies. In addition, PET/CT appeared to show clear advantage over CT and PET alone for the diagnosis of BC recurrence. Future research should include: prospective studies with patient populations clearly defined with regard to their clinical presentation; a study of diagnostic accuracy of PET/CT compared with conventional imaging techniques; a study of PET/CT compared with whole-body MRI; studies investigating the possibility of using PET/CT as a replacement for rather than an addition to CITs; and using modelling of the impact of PET/CT on patient outcomes to inform the possibility of conducting large-scale intervention trials.


A New Method for a Virtue-Based Responsible Conduct of Research Curriculum: Pilot Test Results.
Drawing on Pennock's theory of scientific virtues, we are developing an alternative curriculum for training scientists in the responsible conduct of research (RCR) that emphasizes internal values rather than externally imposed rules. This approach focuses on the virtuous characteristics of scientists that lead to responsible and exemplary behavior. We have been pilot-testing one element of such a virtue-based approach to RCR training by conducting dialogue sessions, modeled upon the approach developed by Toolbox Dialogue Initiative, that focus on a specific virtue, e.g., curiosity and objectivity. During these structured discussions, small groups of scientists explore the roles they think the focus virtue plays and should play in the practice of science. Preliminary results have shown that participants strongly prefer this virtue-based model over traditional methods of RCR training. While we cannot yet definitively say that participation in these RCR sessions contributes to responsible conduct, these pilot results are encouraging and warrant continued development of this virtue-based approach to RCR training.


The eating attitudes test: psychometric features and clinical correlates.
Psychometric and clinical correlates of the Eating Attitudes Test (EAT) are described for a large sample of female anorexia nervosa (N = 160) and female comparison (N = 140) subjects. An abbreviated 26-item version of the EAT (EAT-26) is proposed, based on a factor analysis of the original scale (EAT-40). The EAT-26 is highly correlated with the EAT-40 (r = 0.98) and the three factors form subscales which are meaningfully related to bulimia, weight, body-image variables and psychological symptoms. Whereas there are no differences between bulimic and restricter anorexia nervosa patients on the total EAT-26 and EAT-40 scores, these groups do indicate significant differences on EAT-26 factors. Norms for the anorexia nervosa and female comparison subjects are presented for the EAT-26, EAT-40 and the EAT-26 factors. It is concluded that the EAT-26 is a reliable, valid and economical instrument which may be useful as an objective measure of the symptoms of anorexia nervosa.


Three distinct clades of cultured heterocystous cyanobacteria constitute the dominant N2-fixing members of biological soil crusts of the Colorado Plateau, USA.
The identity of the numerically dominant N(2)-fixing bacteria in biological soil crusts of the Colorado Plateau region and two outlying areas was determined using multiple approaches, to link the environmental diversity of nifH gene sequences to cultured bacterial isolates from the regions. Of the nifH sequence-types detected in soil crusts of the Colorado Plateau, 89% (421/473) were most closely related to nifH signature sequences from cyanobacteria of the order Nostocales. N(2)-fixing cyanobacterial strains were cultured from crusts and their morphotypes, 16S rRNA gene and nifH gene sequences were characterized. The numerically dominant diazotrophs in the Colorado Plateau crusts fell within three clades of heterocystous cyanobacteria. Two clades are well-represented by phylogenetically and morphologically coherent strains, corresponding to the descriptions of Nostoc commune and Scytonema hyalinum, which are widely recognized as important N(2)-fixing components of soil crusts. A third, previously-overlooked clade was represented by a phylogenetically coherent but morphologically diverse group of strains that encompass the morphogenera Tolypothrix and Spirirestis. Many of the strains in each of these groups contained at least two nifH copies that represent different clusters in the nifH environmental survey.