Identification of a small molecule inhibitor of importin β mediated nuclear import by confocal on-bead screening of tagged one-bead one-compound libraries.
In eukaryotic cells, proteins and RNAs are transported between the nucleus and the cytoplasm by nuclear import and export receptors. Over the past decade, small molecules that inhibit the nuclear export receptor CRM1 have been identified, most notably leptomycin B. However, up to now no small molecule inhibitors of nuclear import have been described. Here we have used our automated confocal nanoscanning and bead picking method (CONA) for on-bead screening of a one-bead one-compound library to identify the first such import inhibitor, karyostatin 1A. Karyostatin 1A binds importin β with high nanomolar affinity and specifically inhibits importin α/β mediated nuclear import at low micromolar concentrations in vitro and in living cells, without perturbing transportin mediated nuclear import or CRM1 mediated nuclear export. Surface plasmon resonance binding experiments suggest that karyostatin 1A acts by disrupting the interaction between importin β and the GTPase Ran. As a selective inhibitor of the importin α/β import pathway, karyostatin 1A will provide a valuable tool for future studies of nucleocytoplasmic trafficking.


Use of human MonoMac6 cells for development of in vitro assay predictive of adjuvant safety in vivo.
Subunit vaccines composed of recombinant or purified antigens have a good safety record but are poorly immunogenic and require adjuvants to activate innate immunity and facilitate antigen specific immune response. Of the many adjuvant formulations that are under development, very few are licensed mainly due to concerns about adverse side effects. The goal of our study was to develop in vitro assays that could predict toxicity of adjuvants in vivo. Pro-inflammatory cytokines IL-β, IL-6, TNF-α, and IL-8 were measured in human primary monocytes and the monocytoid cell line, MonoMac 6 (MM6), activated with a panel of TLR agonists or with adjuvants. A 0.5 EU/ml dose of Standard for endotoxin (previously shown to provide a margin between pyrogenic and non-pyrogenic substances in rabbits) was used as a comparator to establish a "safety threshold". FSL-1, Pam3CSK4, flagellin, and R848 TLR agonists but not Alum, MF59, Poly I:C, or MPL adjuvants induced cytokines in MM6 cells above the safety threshold. To confirm the predictive value of the in vitro assays, FSL-1 and flagellin were injected intramuscularly into New Zealand White (NZW) rabbits. Both TLR agonists induced fever within 6-8h post-injection followed 24-48 h later by increased C reactive protein (CRP). Importantly, an early peak in plasma prostaglandin E2 (PGE(2)) levels preceded rise in body temperature. In vitro production of PGE(2) in monocytes and MM6 cells was found following treatments with various TLR agonists but not with alum, MF59, MPL, or Poly I:C adjuvants. Together, our studies demonstrated a strong correlation between production of pro-inflammatory cytokines above a "safety threshold" and production of PGE(2)in vitro and an increase in body temperature in rabbits. The developed human cell based assays could provide an important tool for early screening of new molecular moieties and adjuvant formulations and may assist in selection of safer products.


Effect of furosemide in canine low-pressure pulmonary edema.
We studied the effect of furosemide on pulmonary oxygen exchange, lung liquid, and central hemodynamics in dogs with pulmonary capillary leak induced by intravenous oleic acid (OA). 2 h after OA, triple indicator-dilution lung liquid volume and pulmonary shunt (Qs/Qt) doubled despite normal pulmonary capillary wedge pressure in 16 dogs compared with dogs not given OA in which no variable change during the same time. Six edematous dogs were then treated with furosemide (1 mg/kg), and 2 h later they showed significant reductions in Qs/Qt and lung liquid. In contrast, six other edematous dogs not given furosemide increased Qs/Qt and lung liquid during the same time. The changes in edema after furosemide could not be attributed to altered wedge or colloid osmotic pressures, and similar changes in Qs/Qt and lung liquid with furosemide were observed in four nephrectomized dogs. We conclude that pulmonary vasoactive effects of furosemide account for reduced shunt and edema in canine pulmonary capillary leak. These effects of furosemide differ from those in cardiogenic pulmonary edema, and suggest a different rationale for diuretic therapy in low-pressure pulmonary edema. Analysis of count rates from 51Cr-labeled erythrocytes and 125I-labeled albumin in lungs excised from 12 dogs indicated that the composition of excess lung liquid did not change with furosemide, and was 50% plasma, 25% blood, and 25% crystalloid.


A model for all genomes: the role of transcription factories.
A model for all genomes involving one major architectural motif is presented: DNA or chromatin loops are tethered to "factories" through the transcription machinery-a polymerase (active or inactive) or its transcription factors (activators or repressors). These loops appear and disappear as polymerases initiate and terminate (and as factors bind and dissociate), so the structure is ever-changing and self-organizing. This model is parsimonious, detailed (and so easily tested), and incorporates elements found in various other models.


Idiosyncratic Mòjiāng virus attachment glycoprotein directs a host-cell entry pathway distinct from genetically related henipaviruses.
In 2012, cases of lethal pneumonia among Chinese miners prompted the isolation of a rat-borne henipavirus (HNV), Mòjiāng virus (MojV). Although MojV is genetically related to highly pathogenic bat-borne henipaviruses, the absence of a conserved ephrin receptor-binding motif in the MojV attachment glycoprotein (MojV-G) indicates a differing host-cell recognition mechanism. Here we find that MojV-G displays a six-bladed β-propeller fold bearing limited similarity to known paramyxoviral attachment glycoproteins, in particular at host receptor-binding surfaces. We confirm the inability of MojV-G to interact with known paramyxoviral receptors in vitro, indicating an independence from well-characterized ephrinB2/B3, sialic acid and CD150-mediated entry pathways. Furthermore, we find that MojV-G is antigenically distinct, indicating that MojV would less likely be detected in existing large-scale serological screening studies focused on well-established HNVs. Altogether, these data indicate a unique host-cell entry pathway for this emerging and potentially pathogenic HNV.


A systematic review of the management and outcome of toxic epidermal necrolysis treated in burns centres.
Toxic epidermal necrolysis (TEN) is a rare condition characterised by mucocutaneous exfoliation of greater than 30% total body surface area (%TBSA), increasingly being treated in burns centres. The rate of mortality varies significantly in the literature, with recent prospective studies in non-burns centres reporting percentage mortality of approximately 45%. We undertook a systematic review of published studies that included TEN patients treated specifically in burns centres to determine a cumulative mortality rate. Electronic searches of MEDLINE, EMBASE and The Cochrane Library (Issue 4, 2010) databases from 1966 onwards were used to identify English articles related to the treatment of TEN in burns centres. The systematic literature search identified 20 studies which specifically described patients with TEN grater than 30% %TBSA. Treatment regimens varied amongst studies, as did mortality. The overall percentage mortality of the combined populations was 30%. Risk factors commonly described as associated with mortality included age, %TBSA and delay to definitive treatment. The review highlights the variation between principles of treatment and mortality amongst burns centres. It offers a standard that burns centre can use to internationally compare their mortality rates. The review supports the ongoing reporting of outcomes in TEN patients with epidermal detachment greater than 30%.


Ontogenetic variability of Bothrops atrox and Bothrops asper snake venoms from Colombia.
The lancehead snakes Bothrops asper and Bothrops atrox inflict 70-90% of the 3000 bites reported every year in Colombia. In this work, the venoms of B. atrox from Meta (Villavicencio, 33 specimens) and B. asper from Antioquia (San Carlos, 45 specimens), all of them born in captivity, were obtained at different ages (0-6 months; 1, 2 and 3-years old) and compared in terms of their pharmacological and immunochemical characteristics. A conspicuous ontogenetic variability was observed in venom samples from both species. Venoms from newborn and juvenile specimens showed higher lethal, hemorrhagic, edema-forming and coagulant activities, whereas venoms from 3-year old specimens showed higher indirect hemolytic, i.e. phospholipase A2 activity, being more significant in the case of B. asper. SDS-polyacrylamide gel electrophoresis of whole venom for both species evidenced a predominance of high mol. mass bands in the venoms from specimens of <1 year of age, with a change towards bands having lower mol. mass as snakes aged. Gel filtration chromatography showed five peaks in the venoms of B. asper of <6 months and in those from 3-year old specimens. Venom of adult specimens showed a higher number of peaks with indirect hemolytic activity than venom of newborn specimens. Polyvalent antivenom produced in Costa Rica recognized all the bands of both venoms from specimens at all ages tested, when assayed by Western blotting.


Salinity-induced changes in the morphology and major mineral nutrient composition of purslane (Portulaca oleracea L.) accessions.
This study was undertaken to determine the effects of varied salinity regimes on the morphological traits (plant height, number of leaves, number of flowers, fresh and dry weight) and major mineral composition of 13 selected purslane accessions. Most of the morphological traits measured were reduced at varied salinity levels (0.0, 8, 16, 24 and 32 dS m(-1)), but plant height was found to increase in Ac1 at 16 dS m(-1) salinity, and Ac13 was the most affected accession. The highest reductions in the number of leaves and number of flowers were recorded in Ac13 at 32 dS m(-1) salinity compared to the control. The highest fresh and dry weight reductions were noted in Ac8 and Ac6, respectively, at 32 dS m(-1) salinity, whereas the highest increase in both fresh and dry weight was recorded in Ac9 at 24 dS m(-1) salinity compared to the control. In contrast, at lower salinity levels, all of the measured mineral levels were found to increase and later decrease with increasing salinity, but the performance of different accessions was different depending on the salinity level. A dendrogram was also constructed by UPGMA based on the morphological traits and mineral compositions, in which the 13 accessions were grouped into 5 clusters, indicating greater diversity among them. A three-dimensional principal component analysis also confirmed the output of grouping from cluster analysis.


Lost in Translation: Language, Terminology, and Understanding of Penile-Anal Intercourse in an HIV Prevention Trial in South Africa, Uganda, and Zimbabwe.
Despite efforts to use culturally appropriate, understandable terms for sexual behavior in HIV prevention trials, the way in which participants interpret questions is underinvestigated and not well understood. We present findings from qualitative interviews with 88 women in South Africa, Uganda, and Zimbabwe who had previously participated in an HIV prevention trial. Findings suggested that participants may have misinterpreted questions pertaining to penile-anal intercourse (PAI) to refer to vaginal sex from behind and subsequently misreported the behavior. Three key issues emerge from these findings: first, the underreporting of socially stigmatized sexual behaviors due to social desirability bias; second, the inaccurate reporting of sexual behaviors due to miscomprehension of research terms; and third, the ambiguity in vernacular terms for sexual behavior and lack of acceptable terms for PAI in some languages. These findings highlight methodological challenges around developing clear and unambiguous definitions for sexual behaviors, with implications not only for clinical trials but also for clinical practice and sexual risk assessment. We discuss the challenges in collecting accurate and reliable data on heterosexual PAI in Africa and make recommendations for improved data collection on sensitive behaviors.


Reducing racial bias among health care providers: lessons from social-cognitive psychology.
The paper sets forth a set of evidence-based recommendations for interventions to combat unintentional bias among health care providers, drawing upon theory and research in social cognitive psychology. Our primary aim is to provide a framework that outlines strategies and skills, which can be taught to medical trainees and practicing physicians, to prevent unconscious racial attitudes and stereotypes from negatively influencing the course and outcomes of clinical encounters. These strategies and skills are designed to: 1) enhance internal motivation to reduce bias, while avoiding external pressure; 2) increase understanding about the psychological basis of bias; 3) enhance providers' confidence in their ability to successfully interact with socially dissimilar patients; 4) enhance emotional regulation skills; and 5) improve the ability to build partnerships with patients. We emphasize the need for programs to provide a nonthreatening environment in which to practice new skills and the need to avoid making providers ashamed of having racial, ethnic, or cultural stereotypes. These recommendations are also intended to provide a springboard for research on interventions to reduce unintentional racial bias in health care.


Huge variation in obtaining ethical permission for a non-interventional observational study in Europe.
Ethical approval (EA) must be obtained before medical research can start. We describe the differences in EA for an pseudonymous, non-interventional, observational European study. Sixteen European national coordinators (NCs) of the international study on very old intensive care patients answered an online questionnaire concerning their experience getting EA. N = 8/16 of the NCs could apply at one single national ethical committee (EC), while the others had to apply to various regional ECs and/or individual hospital institutional research boards (IRBs). The time between applying for EA and the first decision varied between 7 days and 300 days. In 9/16 informed consent from the patient was not deemed necessary; in 7/16 informed consent was required from the patient or relatives. The upload of coded data to a central database required additional information in 14/16. In 4/16 the NCs had to ask separate approval to keep a subject identification code list to de-pseudonymize the patients if questions would occur. Only 2/16 of the NCs agreed that informed consent was necessary for this observational study. Overall, 6/16 of the NCs were satisfied with the entire process and 8/16 were (very) unsatisfied. 11/16 would welcome a European central EC that would judge observational studies for all European countries. Variations in the process and prolonged time needed to get EA for observational studies hampers inclusion of patients in some European countries. This might have a negative influence on the external validity. Further harmonization of ethical approval process across Europe is welcomed for low-risk observational studies. Getting ethical approval for low-risk, non-interventional, observational studies varies enormously across European countries.


Metal toxicity affects predatory stream invertebrates less than other functional feeding groups.
Ecosystem effects of heavy metals need to be identified for a retrospective risk assessment, and potential impacts need to be predicted for a prospective risk assessment. In this study, we established a strong correlation between the toxic pressure of dissolved metals and invertebrate species. We compiled available data from a wide geographical range of Australian streams that were contaminated with heavy metals [mainly copper (Cu) and zinc (Zn)] and the corresponding invertebrate communities. Heavy metal toxicity is positively related to the proportion of predators within the invertebrate community, represented by the predatorratio, with an effect threshold range of 2.6 μg/L - 26 μg/L for Cu and 62 μg/L - 617 μg/L for Zn. These effect concentrations are in the ranges of the concentrations identified in model ecosystems and other field investigations and are just above the existing guideline limits. Heavy metals also affects the taxa richness negatively. Other community measures, such as the evenness, number of EPT (Ephemeroptera, Plecoptera, and Trichoptera) taxa, SPEcies At Risk (SPEAR)pesticides or SPEARsalinity were relatively poorly correlated with heavy metal toxicity in the streams. Therefore, we suggest applying the predatorratio within the community as a starting point for an indicator of the dissolved metal toxicity, the SPEARmetals.


Current status of hematopoietic stem cell transplantation in Taiwan.
In Taiwan, hematopoietic SCT (HSCT) has been used to treat patients with hematological diseases since 1983. Since then, more than 2200 patients have undergone HSCT in 15 large hospitals. The disease entities included acute leukemia in 37% of cases, non-Hodgkin's lymphoma in 26%, CML in 10%, multiple myeloma in 7% and severe aplastic anemia in 6%. The conditioning regimens used were mainly myeloablative (84% of cases). Non-myeloablative regimens were fludarabine-based. The average age of allogeneic recipients was at least 10 years older than those in the era before their application. The grafts of all patients were derived from peripheral blood in 85% of cases, BM in 13% and cord blood (CB) in 2%. Forty percent of HSCT patients received autologous grafts, whereas more than 25% of allogeneic HSCT patients received grafts from unrelated donors, and overall, there were more than 200 Taiwan HSCT recipients. Currently, CB has been used successfully in pediatric patients with thalassemia major and also in adult patients with hematological malignancy. After transplantation, there was a relatively lower prevalence of acute GVHD. However, a relatively higher proportion of hepatitis B carriers in the recipients had led to a higher incidence of viral reactivation and clinical hepatitis, which was dramatically decreased following lamivudine prophylaxis. In conclusion, HSCT has been successfully adapted to routine clinical care in Taiwan. Several important findings contributing to the progress of HSCT in the past two decades have also been noticed on this island.


Fractal component of variability of heart rate and systolic blood pressure in congestive heart failure.
1. There is a substantial non-harmonic or fractal component to the variability of both heart rate and blood pressure in normal subjects. Heart rate is the more complex of these two signals, with respect to the slope, beta, of the 1/f beta relationship. In congestive heart failure, heart rate spectral power is attenuated, but the fractal and harmonic components of heart rate and systolic blood pressure variability have not been characterized. 2. Two groups, each comprising 20 men, were studied during 15 min of supine rest and spontaneous respiration: one with functional class II-IV heart failure (age 52 +/- 2 years; mean +/- SEM) and a second group of healthy men (age 46 +/- 2 years). 3. Total spectral power for heart rate was significantly reduced in heart failure (P < 0.02), whereas total spectral power for systolic blood pressure was similar in the two groups. In both heart failure and normal subjects, 65-80% of total spectral power in these two signals displayed fractal characteristics. 4. In heart failure, the slope of the 1/f beta relationship for heart rate was significantly steeper than in normal subjects (1.40 +/- 0.08 compared with 1.14 +/- 0.05; P < 0.05), indicating reduced complexity of the fractal component of heart rate variability. There was no significant difference in the 1/f beta slope for systolic blood pressure variability between these two groups, but the blood pressure signals were less complex than heart rate variations in both heart failure (2.31 +/- 0.15; P < 0.006) and normal subjects (2.47 +/- 0.15; P < 0.0001). 5. Parasympathetic nervous system activity, as estimated from heart rate variability was reduced (P < 0.01) in patients with heart failure, whereas trends towards increased sympathetic nervous system activity and decreased non-harmonic power were not significant. 6. The non-harmonic components of cardiac frequency are reduced in heart failure. Non-harmonic power is not attenuated, but the complexity of the heart rate signal is less than in subjects with normal ventricular function. A reduction in parasympathetic modulation appears to contribute to this loss of complexity of heart rate. Consequently, the heart rate signal comes to resemble that of blood pressure. In contrast, the variability and complexity of the systolic blood pressure signal is similar in heart failure and normal subjects. This reduced complexity of heart rate variability may have adverse implications for patients with heart failure.


Development of the Dutch Structure for Integrated Children's Palliative Care.
Children's palliative care (CPC) is gaining attention worldwide, facilitated by the exchange of knowledge during regular specialised congresses. This article describes the developments in the Netherlands over the past 15 years. The Foundation for Children's Palliative Expertise (PAL) was established as a nationwide initiative committed to improving palliative care for children countrywide. This led to the development of the first hospital-based CPC team in 2012, which expanded to a total of seven teams adjacent to children's university hospitals. Regional networks for CPC were developed in parallel to these teams from 2014 onwards. The networks are a collaboration of professionals from different disciplines and organisations, from hospital to homecare, and have covered the aspects of CPC nationally from 2019 onwards. They are connected through the Dutch Knowledge Centre for CPC. This centre was established in 2018 by the PAL Foundation in collaboration with the Dutch Association for Pediatrics. In 2013, the first evidence-based guideline, 'palliative care for children', provided access to knowledge for parents and healthcare providers, and in 2017, a format for an individual palliative care plan was established. Within the Knowledge Centre for CPC, a physician's support centre for dilemma's regarding the end of life of children was set up. The efforts to have children's palliative care embedded in the regular Dutch health care insurance are ongoing.


Direct measurement of particle inertial migration in rectangular microchannels.
Particles traveling at high velocities through microfluidic channels migrate from their starting streamlines due to inertial lift forces. Theories predict different scaling laws for these forces and there is little experimental evidence by which to validate theory. Here we experimentally measure the three dimensional positions and migration velocities of particles. Our experimental method relies on a combination of sub-pixel accurate particle tracking and velocimetric reconstruction of the depth dimension to track thousands of individual particles in three dimensions. We show that there is no simple scaling of inertial forces upon particle size, but that migration velocities agree well with numerical simulations and with a two-term asymptotic theory that contains no unmeasured parameters.


Simple organ cornea culture model for re-epithelialization after in vitro excimer laser ablation.
Most of the in vitro work to characterize the effects of clinical laser surgery on corneal tissues has concentrated on the effects on stromal keratocytes and endothelium with little attention being paid to corneal epithelium. Our purpose is to describe the epithelial healing rates observed in freshly cultured rabbit corneas treated with phototherapeutic keratectomy (PTK). Corneas were placed in a simple organ culture system, with media change every 2 days. A clinical excimer laser was used to perform a 6 mm diameter, 100 microm depth transepithelial PTK on 24 cultured rabbit corneas, 1 day after culture initiation. For each post-treatment day, one experimental and one control cornea were removed from culture and stained with fluorescein, photographed, and fixed for histology. Epithelial defect area was measured with digital imaging software and analyzed statistically to assess the re-epithelialization rate. Control corneas, maintained in culture for 1-4 days, had no epithelial defects. Those corneas treated with PTK exhibited an immediate epithelial defect that slowly healed over 3 days. This was confirmed on histopathological analysis. A significant linear trend in re-epithelialization across the time points studied was found (F = 80.48, P = 0.0029). The slope of the linear regression model showed an estimate rate of re-epithelialization of -6.70 over the 3 days. We have described the development of a simple, whole organ, rabbit cornea culture model for re-epithelialization after PTK. Our rates of epithelial healing resemble those found in the literature in live rabbit models. Therefore, this model may possibly be used to monitor epithelial wound healing in different corneal diseases or injuries.


Efficient and heritable functional knock-out of an adult phenotype in Drosophila using a GAL4-driven hairpin RNA incorporating a heterologous spacer.
We have developed a modified RNA interference (RNAi) method for generating gene knock-outs in Drosophila melanogaster. We used the sequence of the yellow (y) locus to construct an inverted repeat that will form a double-stranded hairpin structure (y-IR) that is under the control of the upstream activating sequence (UAS) of the yeast transcriptional activator GAL4. Hairpins are extremely difficult to manipulate in Escherichia coli, so our method makes use of a heterologous 330 bp spacer encoding sequences from green fluorescent protein to facilitate the cloning steps. When the UAS-y-IR hairpin is expressed under the control of different promoter-GAL4 fusions, a high frequency of y pigment phenocopies is obtained in adults. Consequently this method for producing gene knock-outs has several advantages over previous methods in that it is applicable to any gene within the fly genome, greatly facilitates cloning of the hairpin, can be used if required with GAL4 drivers to avoid lethality or to induce RNAi in a specific developmental stage and/or tissue, is useful for generating knock-outs of adult phenotypes as reported here and, finally, the system can be manipulated to investigate the trans-acting factors that are involved in the RNAi mechanism.


Reconsidering the role of social disadvantage in physical and mental health: stressful life events, health behaviors, race, and depression.
Prevalence of depression is associated inversely with some indicators of socioeconomic position, and the stress of social disadvantage is hypothesized to mediate this relation. Relative to whites, blacks have a higher burden of most physical health conditions but, unexpectedly, a lower burden of depression. This study evaluated an etiologic model that integrates mental and physical health to account for this counterintuitive patterning. The Baltimore Epidemiologic Catchment Area Study (Maryland, 1993-2004) was used to evaluate the interaction between stress and poor health behaviors (smoking, alcohol use, poor diet, and obesity) and risk of depression 12 years later for 341 blacks and 601 whites. At baseline, blacks engaged in more poor health behaviors and had a lower prevalence of depression compared with whites (5.9% vs. 9.2%). The interaction between health behaviors and stress was nonsignificant for whites (odds ratio (OR = 1.04, 95% confidence interval: 0.98, 1.11); for blacks, the interaction term was significant and negative (β: -0.18, P < 0.014). For blacks, the association between median stress and depression was stronger for those who engaged in zero (OR = 1.34) relative to 1 (OR = 1.12) and ≥2 (OR = 0.94) poor health behaviors. Findings are consistent with the proposed model of mental and physical health disparities.


Molecular and physiological responses during thermal acclimation of leaf photosynthesis and respiration in rice.
To further our understanding of how sustained changes in temperature affect the carbon economy of rice (Oryza sativa), hydroponically grown plants of the IR64 cultivar were developed at 30°C/25°C (day/night) before being shifted to 25/20°C or 40/35°C. Leaf messenger RNA and protein abundance, sugar and starch concentrations, and gas-exchange and elongation rates were measured on preexisting leaves (PE) already developed at 30/25°C or leaves newly developed (ND) subsequent to temperature transfer. Following a shift in growth temperature, there was a transient adjustment in metabolic gene transcript abundance of PE leaves before homoeostasis was reached within 24 hr, aligning with Rdark (leaf dark respiratory CO2 release) and An (net CO2 assimilation) changes. With longer exposure, the central respiratory protein cytochrome c oxidase (COX) declined in abundance at 40/35°C. In contrast to Rdark , An was maintained across the three growth temperatures in ND leaves. Soluble sugars did not differ significantly with growth temperature, and growth was fastest with extended exposure at 40/35°C. The results highlight that acclimation of photosynthesis and respiration is asynchronous in rice, with heat-acclimated plants exhibiting a striking ability to maintain net carbon gain and growth when exposed to heat-wave temperatures, even while reducing investment in energy-conserving respiratory pathways.


Noninvasive diagnosis of pulmonary embolism.
Pulmonary embolism (PE), with an incidence of 23 per 100,000 patients per year, is a frequent clinical problem, responsible for 200,000 deaths each year in the United States. Pulmonary angiography, the gold standard for diagnosing PE, is invasive, costly and not universally available. Moreover, PE is confirmed in only approximately 30% of patients in whom it is suspected, rendering noninvasive screening tests necessary. Several strategies have been recently proposed to reduce the need for pulmonary angiography in the diagnostic workup of pulmonary embolism. The objective of this article is to analyze the individual performance of the new diagnostic instruments and their combination in rational diagnostic strategies. The author has been working in this field and has contributed original papers on diagnosis of pulmonary embolism and cost-effectiveness of noninvasive diagnostic tests. In addition, the material examined in this article includes articles published in the journals covered by the Science Citation Index and Medline. Several strategies have been recently proposed to reduce the need for pulmonary angiography in the diagnostic workup of pulmonary embolism. The PIOPED study has established the value of ventilation-perfusion lung scan, a normal perfusion lung scan virtually ruling out PE, whereas a high probability lung scan is considered diagnostic in face of reasonable clinical suspicion. All other lung scan results are nondiagnostic. However, clinical evaluation, although insufficiently accurate to yield a definitive diagnosis, is probably reliable enough to be used for estimating pretest probability of PE. The combination of a low clinical probability of PE and a so-called low probability lung scan yields a very low posttest probability of PE, thus foregoing the need for pulmonary angiography. Other useful instrument in patients with nondiagnostic scans is plasma D-dimer (DD) measurement (ELISA assay), which when under a cutoff value of 500 micrograms/L potentially exclude PE, due to high sensitivity (97%). Conversely, venous compression ultrasonography of the lower limbs (US) is highly specific (98%) for deep vein thrombosis (DVT), and disclosing a DVT warrants anticoagulant treatment without resorting to angiography. The potential role of echocardiography is also discussed. The rational sequence of noninvasive tests is currently under discussion. Performing D-dimer and US before lung scan may be the most cost-effective strategy, pulmonary angiography being performed only in case of an inconclusive noninvasive workup. Even though PE remains a difficult diagnostic challenge, the availability of novel noninvasive tests (plasma D-dimer and ultrasonography of the lower limbs) and the rehabilitation of clinical assessment allow a more rational and sparse prescription of pulmonary angiography. More work needs to be done to assess test performances and refine diagnostic strategies in distinct patient subgroups, particularly those hospitalized. Screening patients with plasma D-dimer and ultrasonography of the lower limbs may be the most cost-effective strategy, at least in outpatients.


The development of spatial and memory circuits in the rat.
We provide a concise review of recent studies related to the development of neural circuits supporting spatial navigation and memory in the rat. We chart the relative timeline of the emergence of the four main classes of spatially tuned neurons within the hippocampus and related limbic areas: head direction cells emerge earliest (postnatal day 12, P12), before the eyes of the rats are even open, followed by place cells and boundary responsive cells; grid cells emerge last, around the age of weaning (P21). The rate of maturation is unique to each type of neuron, with the head direction and grid cells showing rapid developmental spurts, in contrast to place cells, which show a more gradual trend of maturation. Interestingly, the emergence of allocentric spatial abilities occurs only after the full complement of spatial neurons becomes functional at P20-21, whereas associative processing in the place cell network is evident from as early as P16. We also present evidence supporting the view that the sensory inputs, which are particularly salient to adult spatial networks, may not be essential for the immature spatial system. Crucially, visual information, although more salient than other sensory modalities for anchoring the adult head direction system, does not appear to be essential for setting up the immature head direction network. We conclude by highlighting an urgent need for new theoretical models that can account for the sequential emergence of spatial cells, as well as the lack of primacy of vision in the early organization of the head direction network. WIREs Cogn Sci 2017, 8:e1424. doi: 10.1002/wcs.1424 For further resources related to this article, please visit the WIREs website.


Blockade of CD40/CD40 ligand interactions attenuates skin fibrosis and autoimmunity in the tight-skin mouse.
To assess the association of CD40/CD40 ligand (CD40L) interactions with the development of skin fibrosis and autoimmunity in tight-skin (TSK/+) mouse, which is a mouse model for human systemic sclerosis. Newly born TSK/+ mice were treated with murine anti-CD40L monoclonal antibody (100 microg intraperitoneally weekly). Hypodermal thickness of 8-week-old female mice (defined as the thickness of a subcutaneous loose connective tissue layer beneath the panniculus carnosus) was measured under a light microscope. All skin sections were taken from the para-midline, upper back region. Serum anti-topoisomerase I autoantibody levels, serum immunoglobulin levels and plasma soluble CD40L levels were determined by enzyme-linked immunosorbent assay. For analysis of lymphocyte surface molecules, single cell suspensions of lymphocytes were stained by monoclonal antibodies. Proliferation of TSK/+ B cells and fibroblasts to anti-CD40 antibodies was assessed by the uptake of [3H]-labelled thymidine and bromodeoxyuridine, respectively. The blockade of CD40/CD40L interactions by anti-CD40L monoclonal antibody significantly reduced cutaneous fibrosis (65%) and anti-topoisomerase I autoantibody in TSK/+ mice. Anti-CD40L monoclonal antibody also normalised B lymphocyte abnormal activation in TSK/+ mice, demonstrated by hyper-gamma-globulinaemia. Furthermore, augmented CD40/CD40L interactions in TSK/+ mice were suggested by upregulated expression of CD40L on CD4(+) T cells, elevated plasma soluble CD40L levels. The hyperresponsiveness to CD40 stimulation was also observed in TSK/+ B cells and fibroblasts. Cutaneous fibrosis and autoimmunity in TSK/+ mice are closely correlated with CD40/CD40L interactions.


TRB3 is stimulated in diabetic kidneys, regulated by the ER stress marker CHOP, and is a suppressor of podocyte MCP-1.
The prevalence of diabetic nephropathy continues to rise, highlighting the importance of investigating and discovering novel treatment strategies. TRB3 is a kinase-like molecule that modifies cellular survival and metabolism and interferes with signal transduction pathways. Herein, we report that TRB3 expression is increased in the kidneys of type 1 and type 2 diabetic mice. TRB3 is expressed in conditionally immortalized podocytes; however, it is not stimulated by elevated glucose. The diabetic milieu is associated with increased oxidative stress and circulating free fatty acids (FFA). We show that reactive oxygen species (ROS) such as H(2)O(2) and superoxide anion (via the xanthine/xanthine oxidase reaction) as well as the FFA palmitate augment TRB3 expression in podocytes. C/EBP homologous protein (CHOP) is a transcription factor that is associated with the endoplasmic reticulum stress response. CHOP expression increases in diabetic mouse kidneys and in podocytes treated with ROS and FFA. In podocytes, transfection of CHOP increases TRB3 expression, and ROS augment recruitment of CHOP to the proximal TRB3 promoter. MCP-1/CCL2 is a chemokine that contributes to the inflammatory injury associated with diabetic nephropathy. In these studies, we demonstrate that TRB3 can inhibit basal and stimulated podocyte production of MCP-1. In summary, enhanced ROS and/or FFA associated with the diabetic milieu induce podocyte CHOP and TRB3 expression. Because TRB3 inhibits MCP-1, manipulation of TRB3 expression could provide a novel therapeutic approach in diabetic kidney disease.


Serum uric acid level is an independent risk factor for presence of calcium in coronary arteries: an observational case-controlled study.
A link between uric acid levels and cardiovascular diseases has been previously reported. Coronary artery calcium score (CACS) is a marker of atherosclerotic disease and a predictor of cardiovascular events. We sought to determine if serum uric acid level is an independent risk factor for the presence of calcium in coronary arteries. Four hundred and forty-two patients who were evaluated in the cardiology outpatient clinic for suspected coronary heart disease with a low-moderate risk for coronary artery disease were included in this observational case-controlled study. Serum uric acid levels were measured with colorimetric methods. CACS were performed using a 64-slice CT scanner. Patients were divided to 3 groups according to their CACS value (Group 1: CACS=0, Group 2: CACS 1-100, Group 3: CACS>100). The demographical characteristics and laboratory findings of 3 groups were similar, except age, fasting glucose levels and serum uric acid levels. Serum uric acid levels were found to increase significantly with increasing CACS (p=0.001). Patients were grouped according to presence CAC (CACS=0 and CACS≥1) and in the multiple regression analysis, age (OR, 1.11, 95% CI, 1.07-1.16), smoking (OR, 3.83, 95% CI, 2.06-7.09), serum uric acid levels (OR, 1.26, 95% CI, 1.04-1.54) and average 10-year total risk of Framingham risk score (OR, 1.13, 95% CI, 1.04-1.09) appeared as independent factors predictive of presence of CAC (p<0.05). Serum uric acid level is an independent risk factor for presence of coronary calcium. Moreover, increasing levels of serum uric acid are associated with increasing CACS.


ε-Aminocaproic acid and clinical value in cardiac anesthesia.
The primary aim was to compare the "clinical value" of tranexamic acid (TXA) with ε-aminocaproic acid (EACA) when used for blood conservation during high-risk cardiac surgery. Data previously reported by the Blood Conservation Using Antifibrinolytics in a Randomized Trial (BART) study investigators were reanalyzed independently after appropriate statistical adjustment. The authors compared TXA with EACA for important primary and secondary outcomes and applied the "clinical value" equation to this comparison. BART, the largest blinded multicenter study on this topic to date, compared all 3 commonly used antifibrinolytics head-to-head in a randomized dose-equivalent fashion during high-risk cardiac surgery. Comparisons of TXA with EACA with application of the clinical value equation was not performed specifically by the BART investigators. One thousand five hundred fifty patients enrolled in 2 of the 3 arms of the BART study were included in the analysis (TXA, n= 770 and EACA, n = 780, with data reported by the investigators in the New England Journal of Medicine). The major finding was that there were no significant differences in overall safety and clinically important efficacy between TXA and EACA. Considering the substantial difference in costs and with the increasing volume of high-risk cardiac surgery, EACA has increased "clinical value" when compared with TXA. EACA should be the antifibrinolytic medication of choice for high-risk cardiac surgery.


Dynamic viewer of medical events in electronic medical record.
Medical record should enable doctors to comprehend the patient's history and select suitable medical treatment. In paper based medical records, medical events (examination, treatment etc.) are recorded successively, and problem oriented recording is difficult to be applied to patients with much information and a long history. Consequently it is not easy to understand the patient's history from paper based medical records. In order to solve this problem, we developed the flow sheet system in our electronic medical record (EMR). To make a flow sheet, we analyzed the structure of the medical event data. In this paper we introduced the medical event information model for our EMR. Furthermore, we clarified the specification of the data presentation on the flow sheet. We developed the flow sheet on the basis of these analyses. Because there are 3 layers in the vertical axis of the flow sheet, many items of the medical event can be displayed on the screen. When user clicks the cell, the corresponding detail data including images are shown. This system functions to link medical event items with a problem, and shows the bundled items on the flow sheet when the user selects the problem. We implemented this system in Osaka University Hospital. The number of the types of medical events and those of detail events in inpatients are 5.0+1.7 (mean+SD) and 60+47, respectively. The medical doctors in Osaka University Hospital evaluated this system, and concludes that the flow sheet data presentation makes comprehension of the patient's history easier than paper based records. As to the function of bundling the items relevant to the problem, they feel it is especially useful for patients with chronic disease. Thus the flow sheet data presentation in EMR is useful for medical practice.


The Role of lncRNAs in the Pathobiology and Clinical Behavior of Multiple Myeloma.
MM is a hematological neoplasm that is still considered an incurable disease. Besides established genetic alterations, recent studies have shown that MM pathogenesis is also characterized by epigenetic aberrations, such as the gain of de novo active chromatin marks in promoter and enhancer regions and extensive DNA hypomethylation of intergenic regions, highlighting the relevance of these non-coding genomic regions. A recent study described how long non-coding RNAs (lncRNAs) correspond to 82% of the MM transcriptome and an increasing number of studies have demonstrated the importance of deregulation of lncRNAs in MM. In this review we focus on the deregulated lncRNAs in MM, including their biological or functional mechanisms, their role as biomarkers to improve the prognosis and monitoring of MM patients, and their participation in drug resistance. Furthermore, we also discuss the evidence supporting the role of lncRNAs as therapeutic targets through different novel RNA-based strategies.


A sulfur mimic of 1,1-bis(diphenylphosphino)methane: a new ligand opens up.
A simple method for synthesizing diphosphine monosulfide species was developed utilizing lithium sulfide and chlorophosphine starting materials. This afforded 1,1,2,2-tetraphenyldiphosphine monosulfide (1), as well as 1,1,2,2-tetracyclohexyldiphosphine monosulfide (2), which could be used as convenient ligand precursors. Upon addition of 1 or 2 to the ruthenium compound Ru(C5Me5)(cod)Cl, the diphosphine monosulfides rearranged to give bidentate bis(ditertiaryphosphino)thioether ligands in Ru(C5Me5)(PPh2SPPh2)Cl (3) and Ru(C5Me5)(PCy2SPCy2)Cl (4).


Interdependence in Health and Functioning Among Older Spousal Caregivers and Care Recipients.
Older spousal caregiving relationships involve support that may be affected by the health of either the caregiver or care recipient. We conducted a longitudinal analysis using pooled data from 4,632 community-dwelling spousal care recipients and caregivers aged ⩾50 from the 2002 to 2014 waves of the Health and Retirement Study. We specified logistic and negative binomial regression models using lagged predictor variables to assess the role of partner health status on spousal caregiver and care recipient health care utilization and physical functioning outcomes. Care recipients' odds of hospitalization, odds ratio (OR): 0.83, p<.001, decreased when caregivers had more ADL difficulties. When spouses were in poorer versus better health, care recipients' bed days decreased (4.69 vs. 2.54) while caregivers' bed days increased (0.20 vs. 0.96). Providers should consider the dual needs of caregivers caring for care recipients and their own health care needs, in adopting a family-centered approach to management of older adult long-term care needs.


The relative importance of psychosocial factors in arthritis: findings from 10,509 Australian women.
To determine the relative importance of psychosocial factors in arthritis diagnosis in an ageing cohort of Australian women. This study focused on 10,509 women from the 1946-1951 cohort who responded to questions on arthritis in the fifth mailed population-based survey of the Australian Longitudinal Study on Women's Health conducted in 2007. Arthritis was characterised by widespread psychosocial concerns, particularly relating to chronic stress and poor mental health. Univariate analyses revealed that in comparison to women without stress, women with moderate/high stress levels had a 2.5-fold increase in reporting arthritis. Experiencing ongoing negative interpersonal life events concerning illness of a family member/close friend and relationship difficulties was also associated with a 1.4-fold increase in the reporting of arthritis. Likewise, significantly reduced levels of optimism and perceived social support were noted (all associations p<.001). Psychiatric diagnosis was also associated with a two-fold increase in having arthritis (p<.001). Following adjustment for behavioural, demographic and health-related characteristics, anxiety was the only psychosocial factor associated with arthritis (OR=1.4, 95% CI=1.2, 1.7; p<.001). This study examined, epidemiologically, the relative importance of psychosocial factors in arthritis in an ageing cohort of Australian women. The findings from this population-based study indicate that women with arthritis are more likely to report a range of psychosocial-related problems, particularly with regard to chronic stress perception and anxiety. Longitudinal analyses are required to examine the processes by which stress and psychosocial factors may contribute to arthritis risk and poor adaptation in terms of health-related quality of life.


Long-term survival after surgical treatment of metachronous bilateral adrenal metastases of non-small cell lung carcinoma.
Background Although resection of the metastases is the treatment of choice for unilateral solitary adrenal metastasis of non-small cell lung carcinoma (NSCLC), the surgical treatment for bilateral adrenal metastases is quite rare, likely due to the coexistence of multiple synchronous metastases at other sites and/or primary adrenal insufficiency following bilateral adrenalectomy. We herein report a rare case of asynchronous metastasis of NSCLC to the bilateral adrenal glands with long-term survival after bilateral adrenalectomy. Case Report A 70-year-old male underwent right upper lobectomy for lung adenocarcinomaT2aN2M0, stageIIIA following induction chemotherapy. Forty-four months later, right adrenalectomy of a right adrenal tumor was performed, which revealed metastatic lung carcinoma. Following the administration of adjuvant chemotherapy, a metastatic tumor was detected in the left adrenal gland. Although there were no other signs of distant metastasis on radiological examinations, he underwent the chemotherapy due to the risk of adrenal insufficiency. However, on follow-up CT the adrenal lesion was found to have enlarged; therefore, left adrenalectomy was performed. Three years and six months later, he was doing well, with no evidence of recurrence. Conclusions Selected patients with solitary adrenal metastases of NSCLC can benefit from an aggressive treatment approach, even if such metastases are bilateral.


Broad phylogenomic sampling improves resolution of the animal tree of life.
Long-held ideas regarding the evolutionary relationships among animals have recently been upended by sometimes controversial hypotheses based largely on insights from molecular data. These new hypotheses include a clade of moulting animals (Ecdysozoa) and the close relationship of the lophophorates to molluscs and annelids (Lophotrochozoa). Many relationships remain disputed, including those that are required to polarize key features of character evolution, and support for deep nodes is often low. Phylogenomic approaches, which use data from many genes, have shown promise for resolving deep animal relationships, but are hindered by a lack of data from many important groups. Here we report a total of 39.9 Mb of expressed sequence tags from 29 animals belonging to 21 phyla, including 11 phyla previously lacking genomic or expressed-sequence-tag data. Analysed in combination with existing sequences, our data reinforce several previously identified clades that split deeply in the animal tree (including Protostomia, Ecdysozoa and Lophotrochozoa), unambiguously resolve multiple long-standing issues for which there was strong conflicting support in earlier studies with less data (such as velvet worms rather than tardigrades as the sister group of arthropods), and provide molecular support for the monophyly of molluscs, a group long recognized by morphologists. In addition, we find strong support for several new hypotheses. These include a clade that unites annelids (including sipunculans and echiurans) with nemerteans, phoronids and brachiopods, molluscs as sister to that assemblage, and the placement of ctenophores as the earliest diverging extant multicellular animals. A single origin of spiral cleavage (with subsequent losses) is inferred from well-supported nodes. Many relationships between a stable subset of taxa find strong support, and a diminishing number of lineages remain recalcitrant to placement on the tree.


Hospital ventilation simulation for the study of potential exposure to contaminants.
Airflow and ventilation are particularly important in healthcare rooms for controlling thermo-hygrometric conditions, providing anaesthetic gas removal, diluting airborne bacterial contamination and minimizing bacteria transfer airborne. An actual hospitalization room was the investigate case study. Transient simulations with computational fluid dynamics (CFD), based on the finite element method (FEM) were performed to investigate the efficiency of the existing heating, ventilation and air-conditioning (HVAC) plant with a variable air volume (VAV) primary air system. Solid modelling of the room, taking into account thermo-physical properties of building materials, architectural features (e.g., window and wall orientation) and furnishing (e.g., beds, tables and lamps) arrangement of the room, inlet turbulence high induction air diffuser, the return air diffusers and two patients lying on two parallel beds was carried out. Multiphysics modelling was used: a thermo-fluidynamic model (convection-conduction and incompressible Navier-Stokes) was combined with a convection-diffusion model. Three 3D models were elaborated considering different conditions/events of the patients (i.e., the first was considered coughing and/or the second breathing). A particle tracing and diffusion model, connected to cough events, was developed to simulate the dispersal of bacteria-carrying droplets in the isolation room equipped with the existing ventilation system. An analysis of the region of droplet fallout and the dilution time of bacteria diffusion of coughed gas in the isolation room was performed. The analysis of transient simulation results concerning particle path and distance, and then particle tracing combined with their concentration, provided evidence of the formation of zones that should be checked by microclimatic and contaminant control. The present study highlights the fact that the CFD-FEM application is useful for understanding the efficiency, adequacy and reliability of the ventilation system, but also provides important suggestions for controlling air quality, patients' comfort and energy consumption in a hospital.


The effect of oil sands process-affected water and model naphthenic acids on photosynthesis and growth in Emiliania huxleyi and Chlorella vulgaris.
Naphthenic acids (NAs) are among the most toxic organic pollutants present in oil sands process waters (OSPW) and enter marine and freshwater environments through natural and anthropogenic sources. We investigated the effects of the acid extractable organic (AEO) fraction of OSPW and individual surrogate NAs, on maximum photosynthetic efficiency of photosystem II (PSII) (FV/FM) and cell growth in Emiliania huxleyi and Chlorella vulgaris as representative marine and freshwater phytoplankton. Whilst FV/FM in E. huxleyi and C. vulgaris was not inhibited by AEO, exposure to two surrogate NAs: (4'-n-butylphenyl)-4-butanoic acid (n-BPBA) and (4'-tert-butylphenyl)-4-butanoic acid (tert-BPBA), caused complete inhibition of FV/FM in E. huxleyi (≥10 mg L(-1)n-BPBA; ≥50 mg L(-1)tert-BPBA) but not in C. vulgaris. Growth rates and cell abundances in E. huxleyi were also reduced when exposed to ≥10 mg L(-1)n- and tert-BPBA; however, higher concentrations of n- and tert-BPBA (100 mg L(-1)) were required to reduce cell growth in C. vulgaris. AEO at ≥10 mg L(-1) stimulated E. huxleyi growth rate (p ≤ 0.002), yet had no apparent effect on C. vulgaris. In conclusion, E. huxleyi was generally more sensitive to NAs than C. vulgaris. This report provides a better understanding of the physiological responses of phytoplankton to NAs which will enable improved monitoring of NA pollution in aquatic ecosystems in the future.


Morphofunctional aspects of the blood-brain barrier.
The blood-brain barrier (BBB) selectively controls the homeostasis of the Central Nervous System (CNS) environment by the specific structural and biochemical features of the endothelial cells, pericytes and glial endfeet, which represent the cellular components of the mature BBB. Endothelial tight junctions (TJs) are the most important structural component of the BBB, and molecular alteration in the phosphorylation state of some TJs proteins, like ZO-1 or occludin, are crucial in determining alterations in the control of BBB vascular permeability. Astrocytes endfeet enveloping the vessels wall, are considered important in the induction and maintenance of the BBB, through secretion of soluble factors, which modulate the expression of enzymatic complexes and antigens by endothelial cells and TJs - associated proteins. Moreover, astrocytes control water flux at BBB site by expressing a specific water channel, namely aquaporin-4 (AQP4), involved in the molecular composition of the orthogonal particles arrays (OAPs) on the perivascular glial endfeet and tightly coupled with the maintenance of the BBB integrity. Disruption of the BBB is a consistent event occurring in the development of several CNS diseases, including demyelinating lesions in the course of relapsing multiple sclerosis, stroke, Duchenne muscular dystrophy (DMD), but also mechanical injures, neurological insults, septic encephalopathy, brain tumors, permanent ischemia or transient ischemia followed by reperfusion. In most cases, these pathological conditions are associated with an increase in microvascular permeability, vasogenic edema, swollen atrocyte endfeet, and BBB disruption.


Myeloid-epithelial cross talk coordinates synthesis of the tissue-protective cytokine leukemia inhibitory factor during pneumonia.
In bacterial pneumonia, lung damage resulting from epithelial cell injury is a major contributor to the severity of disease and, in some cases, can lead to long-term sequelae, especially in the setting of severe lung injury or acute respiratory distress syndrome. Leukemia inhibitory factor (LIF), a member of the IL-6 cytokine family, is a critical determinant of lung tissue protection during pneumonia, but the cellular sources of LIF and the signaling pathways leading to its production in the infected lung are not known. Here, we demonstrate that lung epithelium, specifically alveolar type II cells, is the predominant site of LIF transcript induction in pneumonic mouse lungs. Epithelial cell cultures were induced to express LIF by bacteria and by sterile bronchoalveolar lavage fluid from pneumonic mice. Reciprocal bone marrow chimera studies demonstrated that LIF deficiency in the nonhematopoietic compartment, but not LIF deficiency in hematopoietic cells, eliminated LIF induction during pneumonia. Although NF-κB RelA (p65) is essential for the expression of many cytokines during pneumonia, its targeted mutation in the lung epithelium was inconsequential for pneumonia-driven LIF induction. However, maximal expression of this epithelial-derived cytokine was dependent on NF-κB RelA in myeloid cells. Overall, our data suggest a signaling axis whereby activation of NF-κB RelA in myeloid cells promotes epithelial LIF induction during lung infections, representing a means through which these two cell types collaborate to improve tissue resilience during pneumonia.


Hepatitis E virus infection as a marker for contaminated community drinking water sources in Tibetan villages.
In April-May 2001, a study was conducted to determine the prevalence of antibodies against hepatitis E virus (HEV) among 426 persons 8-49 years of age randomly selected from two groups of rural villages in central Tibet. Group 1 villages were assessed in 1998 as having poor quality water sources; new water systems were then constructed prior to this study. Group 2 villages had higher quality water and were not designated as priority villages for new systems prior to the study. No participants tested positive for IgM; only IgG was detected in the analyzed samples. Overall, 31% of the participants had ever been infected with HEV (95% confidence interval [CI] = 26.7-35.7%). The rate was higher in men (36.6%) than women (26.3%) and highest in those 30-39 years of age (49.1%). The rate of past infection was higher in group 1; the risk ratio was 2.77 (95% CI = 1.98-3.88). This difference is most likely the result of the poor quality of the original water sources in these villages. In resource-poor countries, HEV may be a useful health indicator reflecting the degree of contamination in village water sources. This may be especially important in rural areas (such as Tibet) where maternal mortality ratios are high because HEV may be an important cause of deaths during pregnancy in disease-endemic areas.


A novel approach to the intraoperative assessment of the uncinate margin of the pancreaticoduodenectomy specimen.
Currently, there is no consensus regarding the pancreaticoduodenectomy (PD) margins examined intraoperatively or the technical protocol for frozen section examination. The aim of this work was to summarize our experience regarding the intraoperative examination of the uncinate margin and to compare it with the published literature. Our local protocol for the intraoperative assessment of the uncinate margin of the PD specimen is described in this article. A PubMed search limited to English language publications using terms along the theme of pancreaticoduodenectomy and margin was performed. Retrieved articles were categorized according to whether they discussed frozen section margin examination. Ten articles published between 1981 and 2005 were retrieved which discussed the intraoperative examination of PD specimens. Of the 10 articles, 5 discussed the intraoperative consultation for diagnostic purposes only, 2 discussed the consultation for both diagnostic purposes and assessment of margins, and 3 discussed intraoperative assessment of margins only. Of the total of five articles that discussed the intraoperative assessment of margins, none detailed the technical protocol for examining the uncinate margin. Our proposed protocol for the intraoperative assessment of the uncinate margin of PD specimens allows for its accurate evaluation and has not been described previously in the English literature.


Ray optical light trapping in silicon microwires: exceeding the 2n(2) intensity limit.
We develop a ray optics model of a silicon wire array geometry in an attempt to understand the very strong absorption previously observed experimentally in these arrays. Our model successfully reproduces the n2 ergodic limit for wire arrays in free space. Applying this model to a wire array on a Lambertian back reflector, we find an asymptotic increase in light trapping for low filling fractions. In this case, the Lambertian back reflector is acting as a wide acceptance angle concentrator, allowing the array to exceed the ergodic limit in the ray optics regime. While this leads to increased power per volume of silicon, it gives reduced power per unit area of wire array, owing to reduced silicon volume at low filling fractions. Upon comparison with silicon microwire experimental data, our ray optics model gives reasonable agreement with large wire arrays (4 μm radius), but poor agreement with small wire arrays (1 μm radius). This suggests that the very strong absorption observed in small wire arrays, which is not observed in large wire arrays, may be significantly due to wave optical effects.


Development and validation of a sport-specific exercise protocol for elite youth soccer players.
The aims of the current study were, firstly, to quantify the motion characteristics of professional youth soccer players and, secondly, to develop and validate a soccer-specific exercise protocol (SSEP). The motion characteristics of 12 first team members and 12 scholars (under 19s), signed to an English Premiership club were determined via motion analysis. Motion profiles from the analysis were then used to develop a SSEP for a non-motorised treadmill. Validity of the protocol was checked with 6, healthy, male soccer players who completed the SSEP and, on a separate occasion, a soccer match. Heart rates were recorded during both trials, in addition, capillary blood and expired air samples were taken, and RPE recorded, during the SSEP. Youth team players covered 10274+/-609 m, compared to 9741+/-882 m by the first team players (t=1.72, p>0.05; 95% CI for the difference = -1174 m to 109 m). The trend for greater mean distance covered by youth players could be attributed to the distances covered while jogging and running. Mean heart rate response was 166+/-9 beatsxmin(-1) during match play and 166+/-12 beatsxmin(-1) during the SSEP (t=0.164, p>0.05). Mean .VO(2) during the SSEP was 70+/-3% of .VO(2max). Blood lactate concentration fell from a mean value of 5.37+/-1.15 mmol x L(-1) during the first half to 4.74+/-1.25 mmol x L(-1) during the 2(nd) half (t=2.52, p<0.05). The findings of this study suggest that the protocol developed induced a similar physiological load to soccer match play and provides the opportunity to study the physiological demands of soccer.


Arbitrary scattering of an electromagnetic zero-order Bessel beam by a dielectric sphere.
Arbitrary electromagnetic (EM) scattering of a zero-order Bessel beam by a homogeneous water sphere in air is investigated. The radial components of the electric and magnetic scattering fields are expressed using a partial wave series involving the beam-shape coefficients, scattering coefficients of the sphere, and half-conical angle of the wavenumber components of the beam. The 3D scattering directivity plots in the far-field region are evaluated using a numerical integration procedure. It is shown here that shifting the sphere off the axis of wave propagation breaks the symmetry in the directivity patterns. Moreover, the scattering strongly depends on the half-cone angle of the beam. This investigation could provide a useful test of finite element codes for the evaluation of EM scattering and radiation forces, which are important in optical tweezers and related particle manipulation applications.


Structural Development Studies of Subtype-Selective Ligands for Peroxisome Proliferator-Activated Receptors (PPARs) Based on the 3,4-Disubstituted Phenylpropanoic Acid Scaffold as a Versatile Template.
Improvements in our understanding of the functions of the nuclear receptor peroxisome proliferator-activated receptor (PPAR) subtypes as pleiotropic regulators of biological responses, including lipid, lipoprotein, glucose homeostasis, inflammation, differentiation and proliferation of various cancer cells, and memory, have provided an opportunity to develop novel PPAR ligands with characteristic subtype selectivity. Such ligands are not only chemical tools to investigate the functions of PPARs, but also candidates for the treatment of PPAR-mediated diseases, including metabolic syndrome, inflammation, dementia, and cancer. This minireview summarizes our work on the design, synthesis, and pharmacological evaluation of subtype-selective PPAR agonists based on the use of 3,4-disubstituted phenylpropanoic acid as a versatile template.


Toxocariasis diagnosed in international travelers at the Institute of Tropical Medicine, Antwerp, Belgium, from 2000 to 2013.
Although infection with Toxocara canis or T. catis (commonly referred as toxocariasis) appears to be highly prevalent in (sub)tropical countries, information on its frequency and presentation in returning travelers and migrants is scarce. In this study, we reviewed all cases of asymptomatic and symptomatic toxocariasis diagnosed during post-travel consultations at the reference travel clinic of the Institute of Tropical Medicine, Antwerp, Belgium. Toxocariasis was considered as highly probable if serum Toxocara-antibodies were detected in combination with symptoms of visceral larva migrans if present, elevated eosinophil count in blood or other relevant fluid and reasonable exclusion of alternative diagnosis, or definitive in case of documented seroconversion. From 2000 to 2013, 190 travelers showed Toxocara-antibodies, of a total of 3436 for whom the test was requested (5.5%). Toxocariasis was diagnosed in 28 cases (23 symptomatic and 5 asymptomatic) including 21 highly probable and 7 definitive. All but one patients were adults. Africa and Asia were the place of acquisition for 10 and 9 cases, respectively. Twelve patients (43%) were short-term travelers (< 1 month). Symptoms, when present, developed during travel or within 8 weeks maximum after return, and included abdominal complaints (11/23 symptomatic patients, 48%), respiratory symptoms and skin abnormalities (10 each, 43%) and fever (9, 39%), often in combination. Two patients were diagnosed with transverse myelitis. At presentation, the median blood eosinophil count was 1720/μL [range: 510-14160] in the 21 symptomatic cases without neurological complication and 2080/μL [range: 1100-2970] in the 5 asymptomatic individuals. All patients recovered either spontaneously or with an anti-helminthic treatment (mostly a 5-day course of albendazole), except both neurological cases who kept sequelae despite repeated treatments and prolonged corticotherapy. Toxocariasis has to be considered in travelers returning from a (sub)tropical stay with varying clinical manifestations or eosinophilia. Prognosis appears favorable with adequate treatment except in case of neurological involvement.


Functional amyloids as inhibitors of plasmid DNA replication.
DNA replication is tightly regulated to constrain the genetic material within strict spatiotemporal boundaries and copy numbers. Bacterial plasmids are autonomously replicating DNA molecules of much clinical, environmental and biotechnological interest. A mechanism used by plasmids to prevent over-replication is 'handcuffing', i.e. inactivating the replication origins in two DNA molecules by holding them together through a bridge built by a plasmid-encoded initiator protein (Rep). Besides being involved in handcuffing, the WH1 domain in the RepA protein assembles as amyloid fibres upon binding to DNA in vitro. The amyloid state in proteins is linked to specific human diseases, but determines selectable and epigenetically transmissible phenotypes in microorganisms. Here we have explored the connection between handcuffing and amyloidogenesis of full-length RepA. Using a monoclonal antibody specific for an amyloidogenic conformation of RepA-WH1, we have found that the handcuffed RepA assemblies, either reconstructed in vitro or in plasmids clustering at the bacterial nucleoid, are amyloidogenic. The replication-inhibitory RepA handcuff assembly is, to our knowledge, the first protein amyloid directly dealing with DNA. Built on an amyloid scaffold, bacterial plasmid handcuffs can bring a novel molecular solution to the universal problem of keeping control on DNA replication initiation.


Glycocalyx breakdown is increased in African children with cerebral and uncomplicated falciparum malaria.
Cerebral malaria (CM) from Plasmodium falciparum infection is associated with endothelial dysfunction and parasite sequestration. The glycocalyx (GCX), a carbohydrate-rich layer lining the endothelium, is crucial in vascular homeostasis. To evaluate the role of its loss in the pathogenesis of pediatric CM, we measured GCX degradation in Tanzanian children with World Health Organization-defined CM (n = 55), uncomplicated malaria (UM; n = 20), and healthy controls (HCs; n = 25). Urine GCX breakdown products [glycosaminoglycans (GAGs)] were quantified using dimethylmethylene blue (DMMB) and liquid chromatography-tandem mass spectrometry assays. DMMB-GAG and mass spectrometry (MS)-GAG (g/mol creatinine) were increased in CM and UM compared with HCs (P < 0.001), with no differences in DMMB-GAG and MS-GAG between CM and UM children or between those with and without a fatal outcome. In CM survivors, urinary GCX DMMB-GAG normalized by d 3. After adjusting for disease severity, DMMB-GAG was significantly associated with parasitemia [partial correlation coefficient (Pcorr) = 0.34; P = 0.01] and plasma TNF (Pcorr = 0.26; P = 0.04) and inversely with plasma and urine NO oxidation products [Pcorr = -0.31 (P = 0.01) and Pcorr = -0.26 (P = 0.03), respectively]. GCX breakdown is increased in children with falciparum malaria, with similar elevations in CM and UM. Endothelial GCX degradation may impair endothelial NO production, exacerbate adhesion-molecule expression, exposure, and parasite sequestration, and contribute to malaria pathogenesis.-Yeo, T. W., Bush, P. A., Chen, Y., Young, S. P., Zhang, H., Millington, D. S., Granger, D. L., Mwaikambo, E. D., Anstey, N. M., Weinberg, J. B. Glycocalyx breakdown is increased in African children with cerebral and uncomplicated falciparum malaria.


Spatial Heterogeneity and Temporal Trends in Malaria on the Thai⁻Myanmar Border (2012⁻2017): A Retrospective Observational Study.
Malaria infections remain an important public health problem for the Thai-Myanmar border population, despite a plan for the elimination by the end of 2026 (Thailand) and 2030 (Myanmar). This study aimed to explore spatiotemporal patterns in Plasmodium falciparum and Plasmodium vivax incidence along the Thai-Myanmar border. Malaria cases among Thai citizens in 161 sub-districts in Thailand's Kanchanaburi and Tak Provinces (2012-2017) were analyzed to assess the cluster areas and temporal trends. Based on reported incidence, 65.22% and 40.99% of the areas studied were seen to be at elimination levels for P. falciparum and P. vivax already, respectively. There were two clear clusters of malaria in the region: One in the northern part (Cluster I), and the other in the central part (Cluster II). In Cluster I, the malaria season exhibited two peaks, while there was only one peak seen for Cluster II. Malaria incidence decreased at a faster rate in Cluster I, with 5% and 4% reductions compared with 4% and 3% reductions in P. falciparum and P. vivax incidence per month, respectively, in Cluster II. The decreasing trends reflect the achievements of malaria control efforts on both sides of the Thai-Myanmar border. However, these clusters could act as reservoirs. Perhaps one of the main challenges facing elimination programs in this low transmission setting is maintaining a strong system for early diagnosis and treatment, even when malaria cases are very close to zero, whilst preventing re-importation of cases.


[Current value of magnetic resonance imaging for diagnosis of coronary heart disease].
Magnetic Resonance Imaging (MRI) is a non-invasive imaging technique with increasing importance in clinical medicine. It has become a valuable and reliable imaging tool in the diagnosis and management of many medical and surgical conditions. Important advantages of MRI are its flexibility in orientation of imaging plane and the possibility of both anatomical and functional imaging. MRI is based on the application and detection of radio signals and works without any exposure to ionizing radiation, and therefore it is regarded as a safe imaging technique. In the heart there are well established imaging indications such as in acquired and congenital heart disease, pericardial and aortic disease and visualisation of cardiac masses and hypertrophy. Its applications in coronary artery disease (CAD) have been relatively limited, but recent developments in ultrafast imaging sequences and computer hardware have led to a considerable improvement in spatial and temporal image resolution. This has made applications in CAD a possibility, particularly coronary imaging and myocardial perfusion imaging. Recent clinical studies report good correlation between Magnetic Resonance Coronary Angiography (MRCA) and conventional x-ray contrast angiography in the detection of coronary lesions. In the assessment of coronary artery bypass graft (CABG) patency and the definition of anomalous coronary arteries, MRI showed good sensitivity and specificity. The first results of coronary artery flow measurements have now been reported. Myocardial perfusion imaging and stress-ventriculography for detection of wall motion abnormalities are reported as indirect imaging methods with high reliability and clinical value in the diagnosis of CAD. This overview describes recent developments in cardiac MRI and assesses the current and future value of MRI for clinical cardiology.


Performance of A-stage process treating combined municipal-industrial wastewater.
A biosorption column and a settling tank were operated for 6 months with combined municipal and industrial wastewaters (1 m3/hr) to study the effect of dissolved oxygen (DO) levels and Fe3+ dosage on removal efficiency of dissolved and suspended organics prior to biological treatment. High DO (>0.4 mg/L) were found to be detrimental for soluble chemical oxygen demand (COD) removals and iron dosing (up to 20 ppm) did not improve the overall performance. The system performed significantly better at high loading rate (>20 kg COD.m-3.d-1) where suspended solids and COD removals were greater than 80% and 60%, respectively. This is a significant improvement compared to the conventional primary sedimentation tank, and the process is a promising alternative for the pre-treatment of industrial wastewater.


Mapping of cosmid clones in Huntington's disease region of chromosome 4.
Huntington's disease (HD) is tightly linked to genetic markers in 4p16.3. We have used a regional somatic cell hybrid mapping panel to isolate and map 25 cosmids to the proximal portion of 4p16.3 and 17 cosmids to the distal portion. The latter were positioned by long-range restriction mapping relative to previously mapped markers. One cosmid, L6 (D4S166), spans the critical breakpoint in the mapping panel that distinguishes proximal and distal 4p16.3. Four of the cosmids mapped distal to D4S90, the previous terminal marker on 4p, and stretched to within 75 kb of the telomere. Several of the cosmids that mapped between L6 and D4S90 were clustered near a number of previously isolated clones in a region with many NotI sites. Cosmid E4 (D4S168) was localized immediately proximal to the one remaining gap in the long-range restriction map of distal 4p16.3. Although pulsed field gel mapping with E4 failed to link the two segments of the map, the intervening gap was excluded as a potential site for the HD gene by genetic analysis.


Optical imaging of SI topography in anesthetized and awake squirrel monkeys.
Orderly topographic maps in the primary somatosensory cortex (SI) serve as an anchor for our understanding of somatosensory cortical organization. However, this view is mostly based on data collected in the anesthetized animal. Less is known about these topographies in the awake primate. Even less is known about the relative activations of different subdivisions of SI (areas 3a, 3b, 1, and 2). Toward the goal of understanding the functional activation of SI, we conducted intrinsic signal optical imaging of areas 3b and 1 in awake squirrel monkeys. Monkeys were imaged repeatedly for a period of >2 years in awake and anesthetized states in response to vibrotactile and electrocutaneous stimuli presented to individual fingerpads. During this period, we found stable somatotopic maps in both the anesthetized and awake states, consistent with electrophysiologically recorded maps in areas 3b and 1 in the anesthetized state. In the awake animal, signal sizes were larger, but variability was greater, leading to decreased signal-to-noise ratios. Topographic activations were larger (in both area and amplitude) in the awake animal, suggesting either a less precise topography and/or more complex integration. This brings into question the role of a precise topographic map during behavior. In addition, whereas in the anesthetized animal strongest imaging signals were obtained from area 3b, in the awake animal, area 1 activation dominated over that in area 3b. Differences in relative dominance of area 3b versus area 1 suggest that inter-areal interactions in the alert animal differ substantially from that in the anesthetized animal.


Treatment of colorectal cancer in older patients: International Society of Geriatric Oncology (SIOG) consensus recommendations 2013.
Colorectal cancer (CRC) is one of the most commonly diagnosed cancers in Europe and worldwide, with the peak incidence in patients >70 years of age. However, as the treatment algorithms for the treatment of patients with CRC become ever more complex, it is clear that a significant percentage of older CRC patients (>70 years) are being less than optimally treated. This document provides a summary of an International Society of Geriatric Oncology (SIOG) task force meeting convened in Paris in 2013 to update the existing expert recommendations for the treatment of older (geriatric) CRC patients published in 2009 and includes overviews of the recent data on epidemiology, geriatric assessment as it relates to surgery and oncology, and the ability of older CRC patients to tolerate surgery, adjuvant chemotherapy, treatment of their metastatic disease including palliative chemotherapy with and without the use of the biologics, and finally the use of adjuvant and palliative radiotherapy in the treatment of older rectal cancer patients. An overview of each area was presented by one of the task force experts and comments invited from other task force members.


Maternal risk factors predicting child physical characteristics and dysmorphology in fetal alcohol syndrome and partial fetal alcohol syndrome.
Previous research in South Africa revealed very high rates of fetal alcohol syndrome (FAS), of 46-89 per 1000 among young children. Maternal and child data from studies in this community summarize the multiple predictors of FAS and partial fetal alcohol syndrome (PFAS). Sequential regression was employed to examine influences on child physical characteristics and dysmorphology from four categories of maternal traits: physical, demographic, childbearing, and drinking. Then, a structural equation model (SEM) was constructed to predict influences on child physical characteristics. Individual sequential regressions revealed that maternal drinking measures were the most powerful predictors of a child's physical anomalies (R² = .30, p < .001), followed by maternal demographics (R² = .24, p < .001), maternal physical characteristics (R²=.15, p < .001), and childbearing variables (R² = .06, p < .001). The SEM utilized both individual variables and the four composite categories of maternal traits to predict a set of child physical characteristics, including a total dysmorphology score. As predicted, drinking behavior is a relatively strong predictor of child physical characteristics (β = 0.61, p < .001), even when all other maternal risk variables are included; higher levels of drinking predict child physical anomalies. Overall, the SEM model explains 62% of the variance in child physical anomalies. As expected, drinking variables explain the most variance. But this highly controlled estimation of multiple effects also reveals a significant contribution played by maternal demographics and, to a lesser degree, maternal physical and childbearing variables.


Addressing the carbon footprint, healthfulness, and costs of self-selected diets in the USA: a population-based cross-sectional study.
The role of diet in health is well established and, in the past decade, more attention has been given to the role of food choices in the environment. The agricultural sector produces about a quarter of the world's greenhouse gas emissions (GHGE), and meat production, especially beef, is an important contributor to global GHGE. Our study aimed to address a fundamental gap in the diet-climate literature: identifying consumers who are receptive to making dietary changes, and the effect of their potential changes on GHGE, diet healthfulness, and diet costs. Dietary data on US individuals from a nationally representative survey were linked to food-related GHGE. We identified individuals receptive to changing their diets (potential changers) as those who reported trying US dietary guidance and were likely to agree that humans contribute to climate change. We assessed GHGE, diet healthfulness measured by the Healthy Eating Index (HEI), and diet costs before and after hypothetical changes replacing either beef or meats with poultry or plant-protein foods. Our sample comprised 7188 individuals, of whom 16% were potential changers. These were disproportionately women, highly educated, or had higher income compared with individuals deemed not likely to change. Replacing 100% of beef intake in potential changers with poultry reduced mean dietary GHGE by 1·38 kg CO2-equivalents per person per day (95% CI 1·19-1·58), a 35·7% decrease. This replacement also increased mean HEI by 1·7% and reduced mean diet costs by 1·7%. We observed the largest changes when replacing all beef, pork, or poultry intake with plant-protein foods (GHGE decreased by 49·6%, mean HEI increased by 8·7%, and dietary costs decreased by 10·5%). Hypothetical replacements in the potential changers alone resulted in whole population reductions in 1-day dietary GHGE of 1·2% to 6·7%, equivalent to 22-126 million fewer passenger vehicle km. Individual-level diet studies that include a variation in response by consumers can improve our understanding of the effects of climate policies such as those that include sustainability information in national dietary guidance. In our study, we found that changes by a small percentage of motivated individuals can modestly reduce the national dietary GHGE. Moreover, these substitutions can modestly improve diet healthfulness and reduce diet costs for individuals who make these changes. Wellcome Trust.


Collective decision-making appears more egalitarian in populations where group fission costs are higher.
Collective decision-making is predicted to be more egalitarian in conditions where the costs of group fission are higher. Here, we ask whether Trinidadian guppies (Poecilia reticulata) living in high or low predation environments, and thereby facing differential group fission costs, make collective decisions in line with this prediction. Using a classic decision-making scenario, we found that fish from high predation environments switched their positions within groups more frequently than fish from low predation environments. Because the relative positions individuals adopt in moving groups can influence their contribution towards group decisions, increased positional switching appears to support the prediction of more evenly distributed decision-making in populations where group fission costs are higher. In an agent-based model, we further identified that more frequent, asynchronous updating of individuals' positions could explain increased positional switching, as was observed in fish from high predation environments. Our results are consistent with theoretical predictions about the structure of collective decision-making and the adaptability of social decision-rules in the face of different environmental contexts.


Levofloxacin disposition in cerebrospinal fluid in patients with external ventriculostomy.
In vitro levofloxacin exhibits both potent or intermediate activity against most of the pathogens frequently responsible for acute bacterial meningitis and synergistic activity with some beta-lactams. Since levofloxacin was shown to penetrate the cerebrospinal fluid (CSF) during meningeal inflammation both in animals and in humans, the disposition of levofloxacin in CSF was studied in 10 inpatients with external ventriculostomy because of communicating hydrocephalus related to subarachnoid occlusion due to cerebral accidents who were treated with 500 mg of levofloxacin intravenously twice a day because of extracerebral infections. Plasma and CSF concentration-time profiles and pharmacokinetics were assessed at steady state. Plasma and CSF levofloxacin concentrations were analyzed by high-pressure liquid chromatography. The peak concentration of levofloxacin at steady state (C(max ss))was 10.45 mg/liter in plasma and 4.06 mg/liter in CSF, respectively, with the ratio of the C(max ss) in CSF to the C(max ss) in plasma being 0.47. The areas under the concentration-time curves during the 12-h dosing interval (AUC(0-tau)s) were 47.69 mg. h/liter for plasma and 33.42 mg. h/liter for CSF, with the ratio of the AUC(0-tau) for CSF to the AUC(0-tau) for plasma being 0.71. The terminal-phase half-life of levofloxacin in CSF was longer than that in plasma (7.02 +/- 1.57 and 5.51 +/- 1.36 h, respectively; P = 0.034). The ratio of the levofloxacin concentration in CSF to the concentration in plasma progressively increased with time, from 0.30 immediately after dosing to 0.99 at the end of the dosing interval. In the ventricular CSF of patients with uninflamed meninges, levofloxacin was shown to provide optimal exposure, which approximately corresponded to the level of exposure of the unbound drug in plasma. The findings provide support for trials of levofloxacin with twice-daily dosing in combination with a reference beta-lactam for the treatment of bacterial meningitis in adults. This cotreatment could be useful both for overcoming Streptococcus pneumoniae resistance and for enabling optimal exposure of the CSF to at least one antibacterial agent for the overall treatment period.


Fluorescence signal enhancement of polydiacetylene vesicle stacks.
Polydiacetylenes provide a good biological/chemical label-free detection system owing to their blue to red color change and fluorescence change from non-fluorescence to red fluorescence. One of the important factors to consider before applying polydiacetylenes to various sensors is to enhance the sensitivity against specific molecules. This paper reports two methods for stacking polydiacetylene vesicles for fluorescence signal enhancement of polydiacetylene on solid substrates. This focuses on how to achieve a highly sensitive sensor chip by introducing two strategies to immobilize the vesicles effectively on substrates. One method is layer-by-layer deposition through reactions of vesicles and avidins alternately on an avidin treated substrate. The other is to prepare a clustered vesicles solution by mixing the appropriate amount of a biotinylated vesicle solution and avidin solution, and then immobilize the clustered vesicles on the substrates. The former allows easy control of the layer thickness, whereas the latter can shorten the process time. These strategies can be applied to a range of biological/chemical sensors for fluorescence signal enhancement.


The risk of birth defects among children of Persian Gulf War veterans.
There has been suspicion that service in the Persian Gulf War affected the health of veterans adversely, and there have been claims of an increased rate of birth defects among the children of those veterans. We evaluated the routinely collected data on all live births at 135 military hospitals in 1991, 1992, and 1993. The data base included up to eight diagnoses from the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) for each birth hospitalization, plus information on the demographic characteristics and service history of the parents. The records of over 75,000 newborns were evaluated for any birth defect (ICD-9-CM codes 740 to 759, plus neoplasms and hereditary diseases) and for birth defects defined as severe on the basis of the specific diagnoses and the criteria of the Centers for Disease Control and Prevention. During the study period, 33,998 infants were born to Gulf War veterans and 41,463 to non-deployed veterans at military hospitals. The overall risk of any birth defect was 7.45 percent, and the risk of severe birth defects was 1.85 percent. These rates are similar to those reported in civilian populations. In the multivariate analysis, there was no significant association for either men or women between service in the Gulf War and the risk of any birth defect or of severe birth defects in their children. This analysis finds no evidence of an increase in the risk of birth defects among the children of Gulf War veterans.


Inhibition of KCa2 and Kv11.1 Channels in Pigs With Left Ventricular Dysfunction.
Inhibition of KCa2 channels, conducting IKCa, can convert atrial fibrillation (AF) to sinus rhythm and protect against its induction. IKCa inhibition has been shown to possess functional atrial selectivity with minor effects on ventricles. Under pathophysiological conditions with ventricular remodeling, however, inhibiting IKCa can exhibit both proarrhythmic and antiarrhythmic ventricular effects. The aim of this study was to evaluate the effects of the IKCa inhibitor AP14145, when given before or after the IKr blocker dofetilide, on cardiac function and ventricular proarrhythmia markers in pigs with or without left ventricular dysfunction (LVD). Landrace pigs were randomized into an AF group (n = 6) and two control groups: SHAM1 (n = 8) and SHAM2 (n = 4). AF pigs were atrially tachypaced (A-TP) for 43 ± 4 days until sustained AF and LVD developed. A-TP and SHAM1 pigs received 20 mg/kg AP14145 followed by 100 µg/kg dofetilide whereas SHAM2 pigs received the same drugs in the opposite order. Proarrhythmic markers such as short-term variability of QT (STVQT) and RR (STVRR) intervals, and the number of premature ventricular complexes (PVCs) were measured at baseline and after administration of drugs. The influence on cardiac function was assessed by measuring cardiac output, stroke volume, and relevant echocardiographic parameters. IKCa inhibition by AP14145 did not increase STVQT or STVRR in any of the pigs. IKr inhibition by dofetilide markedly increased STVQT in the A-TP pigs, but not in SHAM operated pigs. Upon infusion of AP14145 the number of PVCs decreased or remained unchanged both when AP14145 was infused after baseline and after dofetilide. Conversely, the number of PVCs increased or remained unchanged upon dofetilide infusion. Neither AP14145 nor dofetilide affected relevant echocardiographic parameters, cardiac output, or stroke volume in any of the groups. IKCa inhibition with AP14145 was not proarrhythmic in healthy pigs, or in the presence of LVD resulting from A-TP. In pigs already challenged with 100 µg/kg dofetilide there were no signs of proarrhythmia when 20 mg/kg AP14145 were infused. KCa2 channel inhibition did not affect cardiac function, implying that KCa2 inhibitors can be administered safely also in the presence of LV dysfunction.


A 12-week evaluation of annatto tocotrienol supplementation for postmenopausal women: safety, quality of life, body composition, physical activity, and nutrient intake.
Evidence suggests that tocotrienols may benefit bone health in osteopenic women. However, their safety in this population has never been investigated. This study was to evaluate the safety of a 12-week supplementation of annato tocotrienol in postmenopausal osteopenic women, along with effects of the supplementation on quality of life, body composition, physical activity, and nutrient intake in this population. Eighty nine postmenopausal osteopenic women were randomly assigned to 3 treatment arms: (1) Placebo (430 mg olive oil/day), (2) Low tocotrientol (Low TT) (430 mg tocotrienol/day from DeltaGold 70 containing 300 mg tocotrienol) and (3) High tocotrienol (High TT) (860 mg tocotrienol/day from DeltaGold 70 containing 600 mg tocotrienol) for 12 weeks. DeltaGold 70 is an extract from annatto seed with 70% tocotrienol consisting of 90% delta-tocotrienol and 10% gamma-tocotrienol. Safety was examined by assessing liver enzymes (aspartate aminotransferase, alanine aminotransferase), alkaline phosphatase, bilirubin, kidney function (blood urea nitrogen and creatinine), electrolytes, glucose, protein, albumin, and globulin at 0, 6, and 12 weeks. Serum tocotrienol and tocopherol concentrations were assessed and pills counted at 0, 6, and 12 weeks. Quality of life, body composition, physical activity, and dietary macro- and micro-nutrient intake were evaluated at 0 and 12 weeks. A mixed model of repeated measures ANOVA was applied for analysis. Eighty seven subjects completed the study. Tocotrienol supplementation did not affect liver or kidney function parameters throughout the study. No adverse event due to treatments was reported by the participants. Tocotrienol supplementation for 6 weeks significantly increased serum delta-tocotrienol level and this high concentration was sustained to the end of study. There was no difference in serum delta-tocotrienol levels between the Low TT and the High TT groups. No effects of tocotrienol supplementation were observed on quality of life, body composition, physical activity, and nutrient intake. Annatto-derived tocotrienol up to 600 mg per day for 12 weeks appeared to be safe in postmenopausal osteopenic women, particularly in terms of liver and kidney functions. Tocotrienol supplementation for 12 weeks did not affect body composition, physical activity, quality of life, or intake of macro- and micro-nutrients in these subjects. ClinicalTrials.gov identifier: NCT02058420 . Tocotrienols and bone health of postmenopausal women.


Impaired cardiovascular autonomic nervous system function in patients with Churg-Strauss syndrome.
Although peripheral nervous system involvement in patients with Churg-Strauss syndrome (CSS) has been described, little is known about its autonomic part. Autonomic nervous system (ANS) function can be assessed by studying heart rate variability (HRV) and a decrease in the spectrum of HRV correlates with ANS impairment. Out of 24 CSS patients we chose 12 (four males, eight females, aged 40 ± 8.3 years) in disease remission and without cardiac involvement. Twelve age- and sex-matched healthy volunteers served as a control group. All underwent 24-h electrocardiogram (ECG) Holter recordings. HRV was calculated from 1-h segments, including: total power (TP), ultra-low frequency (ULF), very low frequency (VLF), low frequency (LF), and high frequency (HF) powers as well as normalized LF (LF%) and HF (HF%) powers and the LF to HF power ratio (LF/HF). The CSS patients showed decreased HRV parameters in the 1-h domains: TP (2038 vs. 3622 ms(2), p = 0.001), HF (561 vs. 1574 ms(2), p < 0.001), LF (672 vs. 1050 ms(2), p < 0.01), and VLF (544 vs. 738 ms(2), p = 0.016). However, LF% and LF/HF ratio were markedly higher in CSS patients than in controls (53.4% vs. 39%, p < 0.001 and 1.1 vs. 0.64, p < 0.001), whereas HF% was lower in CSS than in controls (46.6% vs. 61%, p < 0.001). These results were independent of duration of the disease, eosinophil count, corticosteroids, or peripheral nerve involvement in the past. The CSS patients show impaired HRV parameters, indicating parasympathetic ANS dysfunction in addition to peripheral nervous system involvement.