Epidermal growth factor mediated healing in stem cell-derived vocal fold mucosa.
The goal of vocal fold wound healing is the reconstitution of functional tissue, including a structurally and functionally intact epithelium. Mechanisms underlying reepithelialization in vocal folds are not known, although it is suspected that healing involves the interplay between several growth factors. We used a three-dimensional human embryonic stem cell-derived model of vocal fold mucosa to examine the effects of one growth factor, exogenous epidermal growth factor (EGF), on wound healing. A scratch wound was created in the in vitro model. Rate of wound healing, epidermal growth factor receptor (EGFR) activation, and cell proliferation after injury were analyzed with and without application of both exogenous EGF and an EGFR inhibitor, gefitinib. Wound repair after injury was significantly hastened by application of exogenous EGF (13.3 μm/h, ± 2.63) compared with absence of exogenous EGF (7.1 μm/h ± 2.84), but inhibited with concurrent addition of Gefitinib (5.2 μm/h, ± 2.23), indicating that EGF mediates wound healing in an EGFR-dependent manner. Immunohistochemistry revealed that EGFR activation occurred only in the presence of exogenous EGF. Although not statistically significant, increased density of Ki67 staining in the epithelium adjacent to the scratch wound was observed after treatment with EGF, suggesting a tendency for exogenous EGF to increase epithelial cell proliferation. Exogenous EGF increases the rate of wound healing in an EGFR-dependent manner in a three-dimensional stem cell-derived model of vocal fold mucosa. This model of wound healing can be used to gain insight into the mechanisms that regulate vocal fold epithelial repair after injury.


Quantitative real-time polymerase chain reaction for malaria diagnosis and its use in malaria vaccine clinical trials.
The demand for an effective malaria vaccine is high, with millions of people being affected by the disease every year. A large variety of potential vaccines are under investigation worldwide, and when tested in clinical trials, researchers need to extract as much data as possible from every vaccinated and control volunteer. The use of quantitative real-time polymerase chain reaction (PCR), carried out in real-time during the clinical trials of vaccines designed to act against the liver stage of the parasite's life cycle, provides more information than the gold standard method of microscopy alone and increases both safety and accuracy. PCR can detect malaria parasites in the blood up to 5 days before experienced microscopists see parasites on blood films, with a sensitivity of 20 parasites/mL blood. This PCR method has so far been used to follow 137 vaccinee and control volunteers in Phase IIa trials in Oxford and on 220 volunteer samples during a Phase IIb field trial in The Gambia.


Bremazocine: a kappa-opioid agonist with potent analgesic and other pharmacologic properties.
Bremazocine is a kappa-opioid receptor agonist with potent analgesic and diuretic activities. As an analgesic it is three- to four-times more potent than morphine, as determined in both hot plate and tail flick tests. Bremazocine and other benzomorphan analogs were synthesized in an effort to produce opiates with greater kappa-opioid receptor selectivity and with minimal morphine-like side effects. Unlike morphine bremazocine is devoid of physical and psychological dependence liability in animal models and produces little or no respiratory depression. While bremazocine does not produce the characteristic euphoria associated with morphine and its abuse, it has been shown to induce dysphoria, a property that limits its clinical usefulness. Similarly to morphine, repeated administration of bremazocine leads to tolerance to its analgesic effect. It has been demonstrated that the marked diuretic effect of bremazocine is mediated primarily by the central nervous system. Because of its psychotomimetic side effects (disturbance in the perception of space and time, abnormal visual experience, disturbance in body image perception, de-personalization, de-realization and loss of self control) bremazocine has limited potential as a clinical analgesic. However, its possible utility for the therapy of alcohol and drug addiction warrants further consideration because of its ability to decrease ethanol and cocaine self-administration in non-human primates. In addition, the ability of bremazocine-like drugs to lower intraocular pressure and to minimize ischemic damage in animal models suggests their possible use in the therapy of glaucoma and cardiovascular disease.


In vitro study on anti-oxidant and anti-inflammatory properties of Varnya Mahakashaya Dashemani (aqueous extract): A polyherbal formulation.
Plants used in Varnya Mahakashaya Dashemani (VMD) formulation were investigated individually by many scientists. Most of them have exhibited antioxidant, anti-inflammatory and antimicrobial activities when they have been extracted with the different solvents. Here, an attempt has been made to analyze these activities in aqueous extract of the whole formulation. The aim of this study was to evaluate antioxidant and anti-inflammatory potential of polyherbal formulation VMD. Phytochemical constituents of VMD extract were analyzed using standardized protocols and Fourier transform infrared spectroscopy analysis for functional groups. The amount of total phenolics and flavonoids was determined using the Folin-Ciocalteau and aluminum chloride method, respectively. The in vitro antioxidant properties of VMD aqueous extract was screened by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. Anti-inflammatory potency was evaluated with inhibition of 15-lipoxygenase (15-LOX). Phytochemical analysis indicated the presence of alkaloids, flavonoids, tannins, saponin and phenols. The total phenolic content of VMD extract was 50 μg/ml of gallic acid equivalent and the total flavonoids content was 90 μg/ml Quercetin equivalent. It showed higher free radicals quenching capacity with an IC50 value of 34.20 ± 3.03 μg/ml for DPPH and ferric reducing ability by FRAP with an equivalent value of 560 μM (Fe++)/g extract. Significant inhibition of 15-LOX enzyme was prominent with increasing concentration of the sample with an IC50 of 33.62 ± 5.8 μg/ml. VMD has high antioxidant, anti-inflammatory potential and further studies can lead to identification and isolation of more potent therapeutic bioactive compounds from this extract.


Deletion mutagenesis identifies a haploinsufficient role for γ-zein in opaque2 endosperm modification.
Quality Protein Maize (QPM) is a hard kernel variant of the high-lysine mutant opaque2. Using γ-irradiation, we created opaque QPM variants to identify opaque2 modifier genes and to investigate deletion mutagenesis combined with Illumina sequencing as a maize (Zea mays) functional genomics tool. A K0326Y QPM deletion mutant was null for the 27- and 50-kD γ-zeins and abolished vitreous endosperm formation. Illumina exon and RNA sequencing revealed a 1.2-megabase pair deletion encompassing the 27- and 50-kD γ-zein genes on chromosome 7 and a deletion of at least 232 kb on chromosome 9. Protein body number was reduced by over 90%, while protein body size is similar to the wild type. Kernels hemizygous for the γ-zein deletion had intermediate 27- and 50-kD γ-zein levels and were semivitreous, indicating haploinsufficiency of these gene products in opaque2 endosperm modification. The γ-zein deletion further increased lysine in QPM in its homozygous and hemizygous states. This work identifies 27-kD γ-zein as an opaque2 modifier gene within the largest QPM quantitative trait locus and may suggest the 50-kD γ-zein also contributes to this quantitative trait locus. It further demonstrates that genome-wide deletions in nonreference maize lines can be identified through a combination of assembly of Illumina reads against the B73 genome and integration of RNA sequencing data.


A Qualitative Study of Barriers to Accessing Water, Sanitation and Hygiene for Disabled People in Malawi.
Globally, millions of people lack access to improved water, sanitation and hygiene (WASH). Disabled people, disadvantaged both physically and socially, are likely to be among those facing the greatest inequities in WASH access. This study explores the WASH priorities of disabled people and uses the social model of disability and the World Health Organization's International Classification of Functioning, Disability and Health (ICF) framework to look at the relationships between impairments, contextual factors and barriers to WASH access. 36 disabled people and 15 carers from urban and rural Malawi were purposively selected through key informants. The study employed a range of qualitative methods including interviews, emotion mapping, free-listing of priorities, ranking, photo voice, observation and WASH demonstrations. A thematic analysis was conducted using nVivo 10. WASH access affected all participants and comprised almost a third of the challenges of daily living identified by disabled people. Participants reported 50 barriers which related to water and sanitation access, personal and hand hygiene, social attitudes and participation in WASH programs. No two individuals reported facing the same set of barriers. This study found that being female, being from an urban area and having limited wealth and education were likely to increase the number and intensity of the barriers faced by an individual. The social model proved useful for classifying the majority of barriers. However, this model was weaker when applied to individuals who were more seriously disabled by their body function. This study found that body function limitations such as incontinence, pain and an inability to communicate WASH needs are in and of themselves significant barriers to adequate WASH access. Understanding these access barriers is important for the WASH sector at a time when there is a global push for equitable access.


Stark-tuned, far-infrared laser for high-frequency plasma diagnostics.
A Stark-tuned optically pumped far-infrared CH(3)OH laser operating at 119 microm has been built. The laser is designed to operate at high power while exhibiting a well-separated Stark doublet. At a pump power of 65 W and an electric field of 1 kV/cm the laser has delivered over 100 mW cw while exhibiting a frequency splitting of 34 MHz. These parameters indicate that this laser would be suitable for use in the present generation of modulated interferometers on large thermonuclear plasma devices. The achieved modulation frequency is more than an order of magnitude higher than could be achieved by using standard techniques.


Diagnosis of aortic interruption by CT angiography.
Interrupted aortic arch (IAA) is a rare congenital malformation of the aortic arch, which might be accompanied with other coexisting cardiovascular anomalies. Many cases with IAA are diagnosed at their neonatal and newborn period but in rare cases the diagnosis is not established until adulthood. The patients may have no clinical symptoms but the signs of heart failure will gradually appear and may cause death. The development of imaging methods such as computed tomography (CT) and magnetic resonance (MR) imaging has dramatically changed the diagnostics. Here we report a 20-year-old young man with IAA associated with sinus venosus atrial septal defect (SVD) and partial anomalous pulmonary venous connection (PAPVC) referred to our hospital.


Intensive pre-Incan metallurgy recorded by lake sediments from the Bolivian Andes.
The history of pre-Columbian metallurgy in South America is incomplete because looting of metal artifacts has been pervasive. Here, we reconstruct a millennium of metallurgical activity in southern Bolivia using the stratigraphy of metals associated with smelting (Pb, Sb, Bi, Ag, Sn) from lake sediments deposited near the major silver deposit of Cerro Rico de Potosí. Pronounced metal enrichment events coincide with the terminal stages of Tiwanaku culture (1000 to 1200 A.D.) and Inca through early Colonial times (1400 to 1650 A.D.). The earliest of these events suggests that Cerro Rico ores were actively smelted at a large scale in the Late Intermediate Period, providing evidence for a major pre-Incan silver industry.


Conductive properties and gating of channels formed by syringopeptin 25A, a bioactive lipodepsipeptide from Pseudomonas syringae pv. syringae, in planar lipid membranes.
Syringopeptin 25A, a pseudomonad lipodepsipeptide, can form ion channels in planar lipid membranes. Pore conductance is around 40 pS in 0.1 M NaCl. Channel opening is strongly voltage dependent and requires a negative potential on the same side of the membrane where the toxin was added. These pores open and close with a lifetime of several seconds. At negative voltages, an additional pore state of around 10 pS and a lifetime of around 30 ms is also present. The voltage dependence of the rates of opening and closing of the stable pores is exponential. This allows estimation of the equivalent charge that is moved across the membrane during the process of opening at about 2.6 elementary charges. When NaCl is present, the pore is roughly 3 times more permeant for anions than for cations. The current voltage characteristic of the pore is nonlinear, i.e., pore conductance is larger at negative than at positive voltages. The maximal conductance of the pore depends on the concentration of the salt present, in a way that varies almost linearly with the conductivity of the solution. From this, an estimate of a minimal pore radius of 0.4 nm was derived.


Pharmacologic activation of a compensatory integrated stress response kinase promotes mitochondrial remodeling in PERK-deficient cells.
The integrated stress response (ISR) comprises the eIF2α kinases PERK, GCN2, HRI, and PKR, which induce translational and transcriptional signaling in response to diverse insults. Deficiencies in PERK signaling lead to mitochondrial dysfunction and contribute to the pathogenesis of numerous diseases. We define the potential for pharmacologic activation of compensatory eIF2α kinases to rescue ISR signaling and promote mitochondrial adaptation in PERK-deficient cells. We show that the HRI activator BtdCPU and GCN2 activator halofuginone promote ISR signaling and rescue ER stress sensitivity in PERK-deficient cells. However, BtdCPU induces mitochondrial depolarization, leading to mitochondrial fragmentation and activation of the OMA1-DELE1-HRI signaling axis. In contrast, halofuginone promotes mitochondrial elongation and adaptive mitochondrial respiration, mimicking regulation induced by PERK. This shows halofuginone can compensate for deficiencies in PERK signaling and promote adaptive mitochondrial remodeling, highlighting the potential for pharmacologic ISR activation to mitigate mitochondrial dysfunction and motivating the pursuit of highly selective ISR activators.


Effects of inhaled prostaglandins E1, E2, and F2alpha on the airway resistance of healthy and asthmatic man.
1. Changes in specific airway conductance after the inhalation of aerosols of prostaglandins (PG) E1, E2, and F2alpha were investigated in healthy and asthmatic subjects. 2. Inhalation of 155 nmol (55 mug) of PGE1 or 156 nmol (55 mug) of PGF2 resulted in consistent minor bronchodilatation in healthy subjects, but in asthmatic patients airway conductance increased significantly, along with subjective improvement. Isoprenaline (988 nmol; 550 mug) inhalation resulted in a similar increase in conductance to that obtained after these two prostaglandins, whereas a control aerosol had no effect. In contrast to the isoprenaline aerosol, both PGE1 and PGF2 were highly irritant to inhale. It was concluded that this made them unsuitable for therapeutic use. 3. Prostaglandin F2alpha inhalation resulted in a dose-related bronchoconstriction in healthy and asthmatic subjects. Asthmatics were approximately 150 than were the healthy subjects but there was very wide and significant variantion in the sensitivity of the asthmatic subjects. In contrast the asthmatic subjects were only 8-5 times more sensitive to histamine than the healthy subjects with less variation in response of individual subjects. The reasons for the hyper-reactivity of asthmatic subjects to PGF2alpha is unknown and no correlation could be drawn between increased sensitivity and age, type of asthma, or treatment. 4. The effects of disodium cromoglycate, flufenamic acid, atropine methonitrate, PGF2 and isoprenaline on PGF2alpha-induced bronchoconstriction were investigated in healthy subjects. Prostaglandin E2 reversed PGF2alpha-induced bronchoconstriction, as did isoprenaline, but prior treatment with the other drugs had no effect in preventing bronchoconstriction.


First dose in neonates: are juvenile mice, adults and in vitro-in silico data predictive of neonatal pharmacokinetics of fluconazole.
Selection of the first-dose-in-neonates is challenging. The objective of this proof-of-concept study was to evaluate a pharmacokinetic bridging approach to predict a neonatal dosing regimen. We selected fluconazole as a paradigm compound. We used data from studies in juvenile mice and adults to develop population pharmacokinetic models using NONMEM. We also develop a physiologically-based pharmacokinetic model from in vitro-in silico data using Simcyp. These three models were then used to predict neonatal pharmacokinetics and dosing regimens for fluconazole. From juvenile mice to neonates, a correction factor of maximum lifespan potential should be used for extrapolation, while a "renal factor" taking into account renal maturation was required for successful bridging based on adult and in vitro-in silico data. Simulations results demonstrated that the predicted drug exposure based on bridging approach was comparable to the observed value in neonates. The prediction errors were -2.2, +10.1 and -4.6 % for juvenile mice, adults and in vitro-in silico data, respectively. A model-based bridging approach provided consistent predictions of fluconazole pharmacokinetic parameters in neonates and demonstrated the feasibility of this approach to justify the first-dose-in-neonates, based on all data available from different sources (including physiological informations, preclinical studies and adult data), allowing evidence-based decisions of neonatal dose rather than empiricism.


Bacteriological profile, antimicrobial susceptibility patterns of the isolates among street vended foods and hygienic practice of vendors in Gondar town, Northwest Ethiopia: a cross sectional study.
There are numerous advantages offered by street vended foods, but evidence exists that foods exposed for sale on the road side may be contaminated by pathogenic microorganisms. However, information on the bacteriological profile, bacterial load and antimicrobial susceptibility patterns of bacterial isolates from street food in Gondar town are lacking. The aim of this study was to assess bacterial profile, bacterial load, and antimicrobial susceptibility patterns of bacterial isolates among street vended foods and also the hygienic practice of vendors in Gondar town, Northwest Ethiopia. Socio-demographic characteristics and the hygienic practices of 24 vendors were collected using structured questionnaire. A total of 72 food samples from four different food items were analyzed and counted by standard aerobic plate count method. Ten grams of each food sample was transferred in to 90 ml of buffered peptone water and homogenized. The homogenates were serially dilute and a volume of 0.1 ml dilution was spread on solid media and incubated at 35-37 °C for 24 h. Antibiotic susceptibility testing was done for isolated species using Muller Hinton agar and data was entered and analyzed by using SPSS version 20.0. Seventy two food samples of street vended food were analysed for bacterial pathogens. 44/72 tested positive, a total of 63 isolates were identified as 19 samples contained two pathogens. The total mean aerobic bacterial count was 6.64 × 104 CFU/g which is varied from 1 × 104-1.86 × 105 CFU/g. S. aureus is the most frequent isolate 34 (53.96%) followed by E.coli 15(23.8%), Enterobacter species 10(15.87%) and Citrobacter species 4(6.3%). Gentamycin, chloramphenicol, tetracycline, ciprofloxacin, and trimethoprim-sulfamethoxazole were found to be the most effective antimicrobials against all isolates but the enterobactereaceae were resistant to ampicillin and Ceftaziidime and S.aureus were resistant to penicillin. The results of this study showed that, the majority of street-vended food items in Gondar were contaminated with one or more different pathogenic bacteria. The presence of these bacteria in foods could lead to potential health problems for consumers. Therefore, health education as well as training in food safety and hygienic handling is required for food handlers to minimize contamination and the likelihood of people falling ill.


Social factors related to risk for violence and sexually transmitted infections/HIV among Asian massage parlor workers in San Francisco.
Asian women who work at massage parlors in San Francisco have high levels of risk for sexually transmitted infections (STIs), including HIV, and being victims of violence, which jeopardizes their health and wellbeing. On the basis of mapping, the targeted districts in San Francisco where massage parlors were located, 23 massage parlors were identified where commercial sex activity took place. Using snowball-sampling methods, 43 Asian female massage parlor workers were recruited for focus groups; 21 participants were Vietnamese and 22 were Thai. Qualitative analyses revealed frequent exposure to violence including verbal or physical abuse from customers and gang members, as well as persistent HIV risk behaviors associated with multiple daily sex partners, inconsistent condom use with customers, and forced sex. Social factors related to gender, immigration status, and socioeconomic status appeared to be closely tied to the health and wellbeing of Asian masseuses. Study findings suggest that individualized as well as community-level interventions are necessary to improve these women's health and decrease their prolonged exposure to risks for STIs and violence.


Detection of cytokines at the site of tuberculin-induced delayed-type hypersensitivity in man.
Cytokines are chiefly local mediators which play an important role in the regulation of the cell-cell interactions which may be involved in the development of the delayed-type hypersensitivity (DTH) reaction. Using immunohistochemical techniques, the presence of IL-1 alpha, IL-1 beta, IL-6, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) in the skin in tuberculin-purified protein derivative (PPD)-induced DTH reactions was investigated in six normal individuals. Cells staining for these cytokines were first observed 6 h after PPD challenge, and they were detected throughout the duration of the 7-day experiment. The number of cells staining for IFN-gamma reached a peak at 48 h, where 33% of the total aggregate cells were positive, but declined thereafter to 3% at day 7. On the other hand, the number of cells staining for TNF-alpha and IL-1 persisted at high levels throughout the observation period of 7 days (e.g. at 48 h and thereafter, about 40% cells positive for TNF-alpha and 20% for IL-1 alpha and IL-1 beta). Double immunofluorescence and staining on sequential sections showed that IFN-gamma-staining cells were CD3+ T cells; TNF-alpha, IL-1 and IL-6 staining cells were mainly of the CD68+ macrophages/monocytes and that 80% of the CD1a+ cells (Langerhans-like cells) in the dermis contained TNF-alpha and IL-1. The presence of these cytokines at the site of inflammation suggests that they may be locally produced by the inflammatory cells. Their persistence during the reaction suggests that they are intimately associated with this response, and are involved in the development of the reaction.


Distribution of positively charged amino acid residues in antimicrobial peptide epinecidin-1 is crucial for in vitro glioblastoma cytotoxicity and its underlying mechanisms.
Epinecidin-1 (epi) was identified from orange-spotted grouper (Epinephelus coioides) and exhibits diverse biological activities. The aims of this study were to investigate how the distribution of positively charged amino acid residues affects epi-mediated cytotoxicity and to examine the molecular mechanism underlying epi-induced cytotoxicity in U87MG human glioblastoma cells. MTS/PMS and trypan blue exclusion assay were used to measure cell viability. Necrotic cell death was confirmed by detecting cyclophilin A release and propidium iodide incorporation. DNA damage was evaluated by measuring phosphorylated H2AX. Intracellular reactive oxygen species (ROS) were analyzed by flow cytometry using dihydroergotamine. Mitochondrial membrane potential was detected by flow cytometry using tetramethylrhodamine, ethyl ester. Overall, we found that epi caused cytotoxicity in U87MG cells by inducing DNA damage and necrosis through mitochondrial hyperpolarization and subsequent ROS production. The proper folding of epi into an α-helical structure was essential for epi-mediated anti-glioblastoma effects. In addition, NFκB signaling was activated in U87MG cells after exposure to epi. Suppression of NFκB further enhanced epi-induced cytotoxicity, ROS generation and DNA damage, indicating that NFκB may play a protective role in epi-induced cytotoxicity. Our findings may be useful for the design and improvement of antimicrobial peptides with anti-cancer activity.


Cardiopulmonary response to oxygen therapy in hypoxaemic chronic airflow obstruction.
The acute change in pulmonary artery pressure in response to oxygen may have prognostic value for patients with chronic obstructive pulmonary disease treated with long term domiciliary oxygen. A study was carried out to elucidate the mechanism of the acute cardio-respiratory response to oxygen in such patients and to determine whether it can be quantified non-invasively. The effects of acute oxygen administration (100% for 20 minutes and 28% oxygen for 24 hours) were assessed by non-invasive means and right heart catheterisation in 17 patients with severe stable hypoxaemic chronic obstructive pulmonary disease. Measurements included change in the ratio of dead space to tidal volume (VD/VT), effective pulmonary capillary blood flow (by rebreathing and single breath soluble gas uptake: QRB, QSB), left ventricular ejection fraction (radionuclide ventriculography), and M mode echocardiographic estimates of ventricular diameters and fractional shortening. These values were compared with those obtained from right heart catheter measurements of pulmonary artery pressure, cardiac index (thermodilution and direct carbon dioxide Fick: QTD, QFICK), and pulmonary vascular resistance. Oxygen administration resulted in a significant fall in pulmonary artery pressure, QTD, and QRB and a significant increase in VD/VT. The fall in QRB after 100% oxygen breathing for 20 minutes correlated strongly with the fall in pulmonary artery pressure (r = 0.86). There was no correlation between the fall in pulmonary artery pressure and the fall in QSB or the risen in VD/VT. Left ventricular ejection fraction did not change significantly. Echocardiography was technically unsatisfactory because of lung hyperinflation. Apart from a possible relation between VO2max and fall in pulmonary artery pressure after 24 hours of 28% oxygen breathing (r = 0.49, p less than 0.1) none of the baseline respiratory function measurements predicted the fall in pulmonary artery pressure or QRB. It is concluded that the cardiopulmonary response to acute oxygen breathing in patients with hypoxic chronic obstructive pulmonary disease includes a reduction in pulmonary artery pressure and cardiac output and a redistribution of pulmonary blood flow, and that rebreathing measurements of effective pulmonary blood flow can be used to quantify this response non-invasively.


The complete mitochondrial genome of the lesser sac-winged bat Saccopteryx leptura (Chiroptera: Emballonuridae) from Costa Rica.
Here we present the first complete mitochondrial genome of the lesser sac-winged bat Saccopteryx leptura (Chiroptera: Emballonuridae) from Costa Rica, assembled from next-generation sequencing data. The mitogenome of Saccopteryx leptura measures 16,577 bp in length, and contains 13 protein-coding genes, 2 ribosomal RNA genes and 22 transfer RNA genes. A slight A + T bias was observed in the mitogenome of Saccopteryx leptura with an overall base composition of 31.5% A, 28.3% T, 25.8% C, and 14.2% G, and a GC content of 40.1%. The gene arrangement was identical to that of previously described bat mitogenomes.


Caloric intake, aging, and mild cognitive impairment: a population-based study.
In a population-based case-control study, we examined whether moderate and high caloric intakes are differentially associated with the odds of having mild cognitive impairment (MCI). The sample was derived from the Mayo Clinic Study of Aging in Olmsted County, Minnesota. Non-demented study participants aged 70-92 years (1,072 cognitively normal persons and 161 subjects with MCI) reported their caloric consumption within 1 year of the date of interview by completing a Food Frequency Questionnaire. An expert consensus panel classified each subject as either cognitively normal or having MCI based on published criteria. We conducted multivariable logistic regression analyses to compute odds ratios (OR) and 95% confidence intervals (95% CI) after adjusting for age, gender, education, depression, medical comorbidity, and body mass index. We also conducted stratified analyses by apolipoprotein E ε4 genotype status. Analyses were conducted in tertiles of caloric intake: 600 to <1,526 kcals per day (reference group); 1,526 to 2,143 kcals per day (moderate caloric intake group); and >2,143 kcals per day (high caloric intake group). In the primary analysis, there was no significant difference between the moderate caloric intake group and the reference group (OR 0.87, 95% CI 0.53-1.42, p = 0.57). However, high caloric intake was associated with a nearly two-fold increased odds of having MCI (OR 1.96, 95% CI 1.26-3.06, p = 0.003) as compared to the reference group. Therefore, high caloric intake was associated with MCI but not moderate caloric intake. This association is not necessarily a cause-effect relationship.


Simulation and analysis of a multi-order imaging Fabry-Perot interferometer for the study of thermospheric winds and temperatures.
We describe an analysis procedure for estimating the thermospheric winds and temperatures from the multi-order two-dimensional (2D) interferograms produced by an imaging Fabry-Perot interferometer (FPI) as imaged by a CCD detector. We also present a forward model describing the 2D interferograms. To investigate the robustness and accuracy of the analysis, we perform several Monte Carlo simulations using this forward model for an FPI that has recently been developed and deployed to northeastern Brazil. The first simulation shows that a slight cross-contamination at high temperatures exists between neighboring orders in the interferogram, introducing a bias in the estimated temperatures and increasing errors in both the estimated temperatures and winds when each order is analyzed in full. The second simulation investigates how using less than an entire order in the analysis reduces the cross contamination observed in the first set of simulations, improving the accuracy of the estimated temperatures. The last simulation investigates the effect of the signal-to-noise ratio on the errors in the estimated parameters. It is shown that, for the specific FPI simulated in this study, a signal-to-noise ratio of 1.5 is required to obtain thermospheric wind errors of 5 m/s and temperature errors of 20 K.


The Manson Impact Structure: 40Ar/39Ar Age and Its Distal Impact Ejecta in the Pierre Shale in Southeastern South Dakota.
The (40)Ar/(39)Ar ages of a sanidine clast from a melt-matrix breccia of the Manson, Iowa, impact structure (MIS) indicate that the MIS formed 73.8 +/- 0.3 million years ago (Ma) and is not coincident with the Cretaceous-Tertiary boundary (64.43 +/- 0.05 Ma). The MIS sanidine is 9 million years older than (40)Ar/(39)Ar age spectra of MIS shock-metamorphosed microcline and melt-matrix breccia interpreted earlier to be 64 to 65 Ma. Grains of shock-metamorphosed quartz, feldspar, and zircon were found in the Crow Creek Member (upper Campanian) at a biostratigraphic level constrained by radiometric ages in the Pierre Shale of South Dakota that are consistent with the (40)Ar/(39)Ar age of 73.8 +/- 0.3 Ma for MIS reported herein.


Total mesorectal excision using a soft and flexible robotic arm: a feasibility study in cadaver models.
Sponsored by the European Commission, the FP7 STIFF-FLOP project aimed at developing a STIFFness controllable Flexible and Learn-able manipulator for surgical operations, in order to overcome the current limitations of rigid-link robotic technology. Herein, we describe the first cadaveric series of total mesorectal excision (TME) using a soft and flexible robotic arm for optic vision in a cadaver model. TME assisted by the STIFF-FLOP robotic optics was successfully performed in two embalmed male human cadavers. The soft and flexible optic prototype consisted of two modules, each measuring 60 mm in length and 14.3 mm in maximum outer diameter. The robot was attached to a rigid shaft connected to an anthropomorphic manipulator robot arm with six degrees of freedom. The controller device was equipped with two joysticks. The cadavers (BMI 25 and 28 kg/m2) were prepared according to the Thiel embalming method. The procedure was performed using three standard laparoscopic instruments for traction and dissection, with the aid of a 30° rigid optics in the rear for documentation. Following mobilization of the left colonic flexure and division of the inferior mesenteric vessels, TME was completed down to the pelvic floor. The STIFF-FLOP robotic optic arm seemed to acquire superior angles of vision of the surgical field in the pelvis, resulting in an intact mesorectum in both cases. Completion times of the procedures were 165 and 145 min, respectively. No intraoperative complications occurred. No technical failures were registered. The STIFF-FLOP soft and flexible robotic optic arm proved effective in assisting a laparoscopic TME in human cadavers, with a superior field of vision compared to the standard laparoscopic vision, especially low in the pelvis. The introduction of soft and flexible robotic devices may aid in overcoming the technical challenges of difficult laparoscopic procedures based on standard rigid instruments.


Stabilization of short telomeres and telomerase activity accompany immortalization of Epstein-Barr virus-transformed human B lymphocytes.
We have measured telomere length and telomerase activity throughout the life span of clones of human B lymphocytes transformed by Epstein-Barr virus. Shortening of telomeres occurred at similar rates in all populations and persisted until chromosomes had little telomeric DNA remaining. At this stage, some of the clones entered a proliferative crisis and died. Only clones in which telomeres were stabilized, apparently by activation of telomerase, continued to proliferate indefinitely, i.e., became immortal. Since loss of telomeres impairs chromosome function, and may thus affect cell survival, we propose that telomerase activity is required for immortality. We have now detected this enzyme in a variety of immortal human cells transformed by different viruses, indicating that telomerase activation may be a common step in immortalization.


Transient, small-scale field-aligned currents in the plasma sheet boundary layer during storm time substorms.
We report on field-aligned current observations by the four Magnetospheric Multiscale (MMS) spacecraft near the plasma sheet boundary layer (PSBL) during two major substorms on 23 June 2015. Small-scale field-aligned currents were found embedded in fluctuating PSBL flux tubes near the separatrix region. We resolve, for the first time, short-lived earthward (downward) intense field-aligned current sheets with thicknesses of a few tens of kilometers, which are well below the ion scale, on flux tubes moving equatorward/earthward during outward plasma sheet expansion. They coincide with upward field-aligned electron beams with energies of a few hundred eV. These electrons are most likely due to acceleration associated with a reconnection jet or high-energy ion beam-produced disturbances. The observations highlight coupling of multiscale processes in PSBL as a consequence of magnetotail reconnection.


[The rapid analysis of polychlorinated quaterphenyls in blood using different diameter capillary column with the high-resolution gas chromatograph high-resolution mass spectrometer].
The polychlorinated quaterphenyl (PCQ) concentrations in blood are important discriminative parameters in yusho patient. In this study, a rapid analytical method for PCQ using different diameter capillary column (rapid-Rtx65TG) with high-resolution gas chromatograph high-resolution mass spectrometer (HRGC/HRMS) instead of the gas chromatograph electron capture detector (ECD/GC) was developed. Using different diameter capillary columns, the analysis time of the HRGC/HRMS was drastically shortened, and the detection sensitivity was improved. In the rapid-Rtx65TG column, a small-bore capillary column (length 1m, I.D. 0.1mm) was connected with the inlet side of the GC, and behind that column, a large-bore capillary column (length 15mm, I.D. 0.53mm) for octadecachloroquaterphenyl (ODCQ) analysis was connected. In the HRGC/HRMS measurement of ODCQ by the rapid-Rtx65TG column, the minimum limit of detection for the apparatus was 0.4 pg, and the minimum limit of determination for the blood was 0.008 ppb. On ECD/GC in the conventional method and HRGC/HRMS in this study, the PCQ concentration in blood including yusho patients and yusho suspected persons was almost equivalent.


The Goldbeter-Koshland switch in the first-order region and its response to dynamic disorder.
In their classical work (Proc. Natl. Acad. Sci. USA, 1981, 78:6840-6844), Goldbeter and Koshland mathematically analyzed a reversible covalent modification system which is highly sensitive to the concentration of effectors. Its signal-response curve appears sigmoidal, constituting a biochemical switch. However, the switch behavior only emerges in the 'zero-order region', i.e. when the signal molecule concentration is much lower than that of the substrate it modifies. In this work we showed that the switching behavior can also occur under comparable concentrations of signals and substrates, provided that the signal molecules catalyze the modification reaction in cooperation. We also studied the effect of dynamic disorders on the proposed biochemical switch, in which the enzymatic reaction rates, instead of constant, appear as stochastic functions of time. We showed that the system is robust to dynamic disorder at bulk concentration. But if the dynamic disorder is quasi-static, large fluctuations of the switch response behavior may be observed at low concentrations. Such fluctuation is relevant to many biological functions. It can be reduced by either increasing the conformation interconversion rate of the protein, or correlating the enzymatic reaction rates in the network.


COL12A1 as a prognostic biomarker links immunotherapy response in breast cancer.
Immunotherapy has shown promising efficacy for breast cancer (BC) patients. Yet the predictive biomarkers for immunotherapy response remain lacking. Based on two GEO datasets, 53 differentially expressed genes associated with durvalumab treatment response were identified. Using least absolute shrinkage and selection operator (LASSO) and univariate Cox regression, four genes (COL12A1, TNN, SCUBE2, and FDCSP) revealed prognostic value in the TCGA BC cohort. COL12A1 outperformed the others, without overlap in its survival curve. Survival analysis by Kaplan-Meier plotter demonstrated that COL12A1 was negatively associated with BC patients' prognosis. A COL12A1-based nomogram was further developed to predict the overall survival in BC patients. The calibration plot revealed an optimal agreement between nomogram prediction and actual observation. Moreover, COL12A1 expression was significantly up-regulated in BC tissues and COL12A1 knockdown impaired the proliferation of MDA-MB-231 and BT549 cells. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment analysis pathway indicated that the function of COL12A1 was related to immunity-related pathways. Immunological analyses illustrated that COL12A1 was correlated with M2 macrophage infiltration and M2 macrophage markers (transforming growth factor beta 1 (TGFB1), interleukin-10, colony stimulating factor 1 receptor (CSF1R) and CD163) in BC. Immunohistochemistry staining further revealed a highly positive relationship of COL12A1 with TGF-β1. The co-incubated models of BC cells and M2 macrophges showed COL12A1 knockdown suppressed M2 macrophage infiltration. Additionally, silencing COL12A1 suppressed TGF-B1 protein expression, and treating with TGFB1 could reverse the inhibitory effects on M2 macrophage infiltration by COL12A1 knockdown. Using immunotherapy datasets, we also found elevated expression of COL12A1 predicted poor response to anti-PD-1/PD-L1 therapy. These results reinforce the current understanding of COL12A1's roles in tumorigenesis and immunotherapy response in BC.


Subjective pulsatile tinnitus cured by carotid endarterectomy. A case report.
A 70-year-old woman with unilateral pulsatile tinnitus was relieved of the complaint after carotid endarterectomy. Internal carotid artery stenosis presenting with pulsatile tinnitus is rare. Endarterectomy can relieve symptoms if the correct diagnosis is made.


[Disclosure of HIV infection from the perspective of adolescents living with HIV/AIDS and their parents and caregivers].
The purpose of this study was to identify relevant issues in the disclosure of HIV-positive status in children and adolescents, aiming to improve the quality of their healthcare. A qualitative study included adolescents living with HIV/ AIDS and their parents and caregivers at AIDS reference services in São Paulo and Santos, Brazil. In-depth interviews and focus group were used. The main reasons for disclosure were: poor treatment adherence, sexual maturity, adolescent's request, and inadequate procedures by medical staff. Disclosure was a critical moment for adolescents, with a strong impact on their life plans and horizons. Adolescents infected through sexual transmission and drug use reported the most problematic scenes involved in disclosure. Despite its initial negative impact, disclosure resulted in improved healthcare and better dialogue among the adolescents, caregivers, and healthcare providers. The adolescents also requested clear, no-nonsense, honest information. The authors conclude that health services need to actively provide an adequate approach to facilitate disclosure of HIV-positive status to children and adolescents, realizing that it is a long-term process supported by the family and a multidisciplinary team.


Naringenin reduces inflammatory pain in mice.
Naringenin is a flavonoid widely consumed by humans that present anti-inflammatory activity and low toxicity. Recently, the analgesic effect of naringenin has been demonstrated in neuropathic pain models. Herein, we tested the analgesic effects of naringenin in several models of inflammatory pain. Mice received treatment with naringenin (16.7-150 mg/kg, per oral), or with the controls anti-inflammatory drugs indomethacin (5 mg/kg, intraperitoneal) or dipyrone (80 mg/kg, intraperitoneal) prior the inflammatory stimuli injection. For acute pain, we used acetic acid- and PBQ-induced visceral pain (abdominal writhings), and formalin-, capsaicin-, and CFA-induced paw flinching and licking. By using an electronic version of von Frey filaments, we also investigated the effects of naringenin in pain intensity to a mechanical stimulus (mechanical hyperalgesia) after carrageenan, capsaicin, CFA, or PGE2 intraplantar injection. Naringenin (50 mg/kg) reduced acute pain behaviors induced by all tested stimuli, including both phases of formalin test, suggesting a direct nociceptor modulatory effect of this compound besides its anti-inflammatory activity. Accordingly, naringenin also inhibited the increased sensitivity to mechanical stimulus induced by carrageenan, capsaicin, and PGE2. Daily treatment with naringenin during 7 days also reduced CFA-induced mechanical hyperalgesia without gastric or hepatic toxicity. The mechanisms of naringenin involve the inhibition of carrageenan-induced oxidative stress, hyperalgesic cytokines (IL-33, TNF-α, and IL-1β) production and NF-κB activation in the paw skin. Naringenin also activated the analgesic NO-cyclic GMP-PKG-ATP sensitive K(+) channel signaling pathway to inhibit carrageenan-induced mechanical hyperalgesia and neutrophil recruitment. These results suggest that naringenin inhibits both inflammatory pain and neurogenic inflammation.


Antiplatelet properties of Pim kinase inhibition are mediated through disruption of thromboxane A2 receptor signaling.
Pim kinases are upregulated in several forms of cancer, contributing to cell survival and tumour development, but their role in platelet function and thrombotic disease has not been explored. We report for the first time that Pim-1 is expressed in human and mouse platelets. Genetic deletion or pharmacological inhibition of Pim kinase results in reduced thrombus formation but is not associated with impaired haemostasis. Attenuation of thrombus formation was found to be due to inhibition of the thromboxane A2 receptor as effects on platelet function was non-additive to inhibition caused by the cyclooxygenase inhibitor indomethacin or thromboxane A2 receptor antagonist GR32191. Treatment with Pim kinase inhibitors caused reduced surface expression of the thromboxane A2 receptor and resulted in reduced responses to thromboxane A2 receptor agonists, indicating a role for Pim kinase in the regulation of thromboxane A2 receptor function. Our research identifies a novel, Pim kinase dependent regulatory mechanism for the thromboxane A2 receptor and represents a new targeting strategy that is independent of COX-1 inhibition or direct antagonism of the thromboxane A2 receptor that whilst attenuating thrombosis does not increase bleeding.


Misoprostol for the treatment of early pregnancy failure.
Nearly 20% of all pregnancies end in early pregnancy failure, and surgical evacuation of retained products of conception is often used to manage this failure. Misoprostol is an inexpensive, stable analog of prostaglandin E(1), and is powerful at contracting the uterus. With intravaginal misoprostol, the peak plasma levels are lower, but the levels after 4 hours are higher, than after oral or sublingual administration. With oral misoprostol, the evacuation rates in early pregnancy varied from about 50% up to 96%. Similar variation in evacuation rates were obtained from small trials with intravaginal misoprostol. To date, only small studies have used sublingual misoprostol, and there has been no direct comparison to oral or intravaginal misoprostol. A recent large clinical trial has shown, that with intravaginal misoprostol 800 microg, an expulsion rate of 84% can be achieved by 8 days. This large trial also established that women prefer misoprostol to surgical evacuation. Two economic evaluations have shown that misoprostol treatment is less costly than surgical intervention. On the basis of recent findings, it seems likely that misoprostol treatment will become a standard or preferred treatment for early pregnancy failure.


Mapping Local and Global Liquid Phase Behavior in Living Cells Using Photo-Oligomerizable Seeds.
Liquid-liquid phase separation plays a key role in the assembly of diverse intracellular structures. However, the biophysical principles by which phase separation can be precisely localized within subregions of the cell are still largely unclear, particularly for low-abundance proteins. Here, we introduce an oligomerizing biomimetic system, "Corelets," and utilize its rapid and quantitative light-controlled tunability to map full intracellular phase diagrams, which dictate the concentrations at which phase separation occurs and the transition mechanism, in a protein sequence dependent manner. Surprisingly, both experiments and simulations show that while intracellular concentrations may be insufficient for global phase separation, sequestering protein ligands to slowly diffusing nucleation centers can move the cell into a different region of the phase diagram, resulting in localized phase separation. This diffusive capture mechanism liberates the cell from the constraints of global protein abundance and is likely exploited to pattern condensates associated with diverse biological processes. VIDEO ABSTRACT.


Effects of form familiarity on perception of words, pseudowords, and nonwords in the two cerebral hemispheres.
Previous investigations of hemispheric processes of word perception provide a mixed picture of the sensitivity of each hemisphere to the familiarity of the visual form of lateralized displays. We investigated this issue by presenting words, pseudowords, and nonwords briefly to either the left (LH) or right (RH) hemisphere in lowercase, uppercase, and a matched, unfamiliar mixed-case form, and used an eye tracker to ensure central fixation and the Reicher-Wheeler task to suppress influences of stimulus asymmetry. Familiarity of form exerted a substantial effect on perception. In particular, perception of LH and RH displays of words, pseudowords, and nonwords was least accurate for mixed case, intermediate for upper case, and most accurate for lowercase. However, form had no effect on the LH advantage observed for words, pseudowords, and nonwords, indicating that form affected processing in both hemispheres to a similar extent. Moreover, LH and RH displays both showed that mixed case disrupted performance most for words, and more for pseudowords than for nonwords, indicating the sensitivity to form shown by each hemisphere reflected more than a general perceptual process. Implications for the role of form familiarity in hemispheric processing of words are discussed.


Cardiac involvement in consecutive unselected hospitalized COVID-19 population: In-hospital evaluation and one-year follow-up.
Cardiovascular disease (CVD) can occur in COVID-19 and has impact on clinical course. Data on CVD prevalence in hospitalized COVID-19 patients and sequelae in survivors is limited. Aim of this prospective study carried out on consecutive unselected COVID-19 population, was to assess: 1) CVD occurrence among hospitalized COVID-19 patients, 2) persistence or new onset of CVD at one-month and one-year follow-up. Over 30 days n = 152 COVID-19 patients underwent cardiovascular evaluation. Standard electrocardiogram (ECG), Troponin and echocardiography were integrated by further tests when indicated. Medical history, arterial blood gas, blood tests, chest computed tomography and treatment were recorded. CVD was defined as the occurrence of a new condition during the hospitalization for COVID-19. Survivors attended a one-month follow-up visit and a one-year telephone follow-up. Forty-two patients (28%) experienced a wide spectrum of CVD with acute myocarditis being the most frequent. Death occurred in 32 patients (21%) and more frequently in patients who developed CVD (p = 0.032). After adjustment for confounders, CVD was independently associated with death occurrence. At one-month follow-up visit, 7 patients (9%) presented persistent or delayed CVD. At one-year telephone follow-up, 57 patients (48%) reported persistent symptoms. Cardiovascular evaluation in COVID-19 patients is crucial since the occurrence of CVD in hospitalized COVID-19 patients is common (28%), requires specific treatment and increases the risk of in-hospital mortality. Persistence or delayed presentation of CVD at 1-month (9%) and persistent symptoms at 1-year follow-up (48%) suggest the need for monitoring COVID-19 survivors.


Morphological and gene expression analysis under cool temperature conditions in rice anther development.
Cool temperature conditions are known to lead to pollen sterility in rice. Pollen sterility is an agriculturally important phenomenon because it imparts a large influence directly on rice yield. However, cool temperature stress tolerance varies among rice cultivars and avoidance of cool temperature stress is difficult by practical method of agriculture. In this study using two rice cultivars, Hitomebore (high tolerance) and Sasanishiki (low tolerance), we analyzed morphological features and gene expression profiles, under cool temperature stress, in anther development of rice. Hitomebore was given cool temperature stress (19 degrees C) at flowering stage, and showed 87.3% seed fertility. Meanwhile, the seed fertility decreased to 41.7% in the case of Sasanishiki. A transverse section of Hitomebore anther revealed that the degradation of the tapetum started at the uninucleate microspore stage, and the tapetum had completely vanished at mature stage. The tapetum provides nutrients for pollen development, and its degradation occurs at a late stage in pollen development. In contrast, degradation of the tapetum did not occur at the uninucleate microspore stage in Sasanishiki, and the tapetum was clearly intact at mature stage, suggesting that tapetum degradation is critical for accurate pollen development and cool temperature tolerance correlated with the degree of tapetum degeneration. In gene expression analysis of anther, 356 genes that showed different expression levels between two cultivars at cool temperatures were found. These genes will lead to understanding the mechanism of cool temperature stress response in rice pollen development and the identification of genes involved in accurate tapetum degradation.


Reconstitution of water channel function of an aquaporin overexpressed and purified from Pichia pastoris.
The aquaporin PM28A is one of the major integral proteins in spinach leaf plasma membranes. Phosphorylation/dephosphorylation of Ser274 at the C-terminus and of Ser115 in the first cytoplasmic loop has been shown to regulate the water channel activity of PM28A when expressed in Xenopus oocytes. To understand the mechanisms of the phosphorylation-mediated gating of the channel the structure of PM28A is required. In a first step we have used the methylotrophic yeast Pichia pastoris for expression of the pm28a gene. The expressed protein has a molecular mass of 32462 Da as determined by matrix-assisted laser desorption ionization-mass spectrometry, forms tetramers as revealed by electron microscopy and is functionally active when reconstituted in proteoliposomes. PM28A was efficiently solubilized from urea- and alkali-stripped Pichia membranes by octyl-beta-D-thioglucopyranoside resulting in a final yield of 25 mg of purified protein per liter of cell culture.


Life cycle assessment of pig slurry treatment technologies for nutrient redistribution in Denmark.
Animal slurry management is associated with a range of impacts on fossil resource use and the environment. The impacts are greatest when large amounts of nutrient-rich slurry from livestock production cannot be adequately utilised on adjacent land. To facilitate nutrient redistribution, a range of different technologies are available. This study comprised a life cycle assessment of the environmental impacts from handling 1000 kg of pig slurry ex-animal. Application of untreated pig slurry onto adjacent land was compared with using four different treatment technologies to enable nutrient redistribution before land application: (a) separation by mechanical screw press, (b) screw press separation with composting of the solid fraction, (c) separation by decanter centrifuge, and (d) decanter centrifuge separation with ammonia stripping of the liquid fraction. Emissions were determined based on a combination of values derived from the literature and simulations with the Farm-N model for Danish agricultural and climatic conditions. The environmental impact categories assessed were climate change, freshwater eutrophication, marine eutrophication, terrestrial acidification, natural resource use, and soil carbon, nitrogen and phosphorus storage. In all separation scenarios, the liquid fraction was applied to land on the pig-producing (donor) farm and the solid fraction transported to a recipient farm and utilised for crop production. Separation, especially by centrifuge, was found to result in a lower environmental impact potential than application of untreated slurry to adjacent land. Composting and ammonia stripping either slightly increased or slightly decreased the environmental impact potential, depending on the impact category considered. The relative ranking of scenarios did not change after a sensitivity analysis in which coefficients for field emissions of nitrous oxide, ammonia and phosphorus were varied within the range cited in the literature. Therefore, the best technology to implement in a given situation depends on the environmental problem in question, local policy, cost and practicality.


Assessment of comparative pain relief and tolerability of SKI306X compared with celecoxib in patients with rheumatoid arthritis: a 6-week, multicenter, randomized, double-blind, double-dummy, phase III, noninferiority clinical trial.
SKI306X, which consists of biologically active ingredients from Clematis mandsburica, Tricbosantbes kirilowii, and Prunella vulgaris, was developed and tested in preclinical trials in Korea. Those studies found that SKI306X was associated with an anti-inflammatory and analgesic effect, and that it can delay the destruction of cartilage in rheumatoid arthritis (RA). The aim of this study was to compare the pain relief and tolerability of SKI306X and celecoxib in patients with RA. This study was a 6-week, multicenter, randomized, double-blind, double-dummy, Phase III, noninferiority clinical trial. Eligible patients were aged 18 to 80 years, had a history of RA with a disease duration of > or =3 months, and were functional American College of Rheumatology (ACR) class I, II, or III before entry. After a washout period of 2 weeks, patients were randomized to SKI306X 200 mg TID or celecoxib 200 mg BID for 6 weeks. The primary end point was a change in patient assessment of pain intensity using a visual analog scale (VAS). The secondary end points were a 20% improvement in response rate as defined by the ACR (ACR20) and the frequency of rescue medication use. Results after 3 and 6 weeks of treatment were compared with baseline and between treatment groups, and all patients were assessed for adverse events (AEs), clinical laboratory data, and vital signs. AEs were identified based on spontaneous reports by patients during interviews conducted by the investigators and the study coordinator. Two hundred twenty-two Korean patients from 7 medical centers were assessed and 183 were enrolled and randomized to 1 of 2 treatment groups. Ninety-one patients (10 male, 81 female; mean [SD] age, 52.13 [12.64] years; mean [SD] duration of RA, 9.08 [10.23] years; no. [%] of ACR class I, II, and III, 13 [14.29], 44 [48.35] and 34 [37.36] patients, respectively) received SKI306X 200 mg TID and 92 patients (10 male, 82 female; mean [SD] age, 51.78 [10.94] years; mean [SD] duration of RA, 8.78 [7.78] years; no. [%] of ACR class I, II, and III, 14 [15.22], 44 [47.83], and 34 [36.96] patients, respectively) received celecoxib 200 mg BID. An analysis of the change in reported pain intensity as determined by VAS (mm) score between baseline and week 3 (mean [SD], 13.64 [16.62] vs 14.45 [15.89]), and between baseline and week 6 (18.4 [20.8] vs 17.9 [19.1], respectively) suggested that SKI306X was not inferior to celecoxib. The number of patients who achieved ACR20 response rate was not significantly different between the SKI306X group and the celecoxib group at week 3 (16/87 [18.4%] vs 24/87 [27.6%], respectively) and at week 6 (29/87 [33.3%] vs 29/87 [33.3%]). The frequency of rescue medication use was not significantly different between the SKI306X group and celecoxib group at week 3 (54/87 [62.1%] vs 47/87 [54.0%], respectively) or week 6 (57/87 [65.5%] vs 49/87 [56.3%]). Drug-related AEs were reported by 27 (29.7%) patients in the SKI306X group and 22 (23.9%) patients in the celecoxib group. The most frequent drug-related AEs were epigastric pain (9/91 [9.9%]) in the SKI306X group and glutamyltranferase elevation (4/92 [4.3%]) in the celecoxib group. No significant between-group differences were observed in the prevalence of drug-related clinical- or laboratory-determined AEs. The results of this study suggest that SKI306X was generally well tolerated and not inferior to celecoxib in regard to pain relief in these Korean patients with RA.


Identification of H5N1-specific T-cell responses in a high-risk cohort in vietnam indicates the existence of potential asymptomatic infections.
Most reported human H5N1 viral infections have been severe and were detected after hospital admission. A case ascertainment bias may therefore exist, with mild cases or asymptomatic infections going undetected. We sought evidence of mild or asymptomatic H5N1 infection by examining H5N1-specific T-cell and antibody responses in a high-risk cohort in Vietnam. Peripheral blood mononuclear cells were tested using interferon-γ enzyme-linked immunospot T assays measuring the response to peptides of influenza H5, H3, and H1 hemagglutinin (HA), N1 and N2 neuraminidase, and the internal proteins of H3N2. Horse erythrocyte hemagglutination inhibition assay was performed to detect antibodies against H5N1. Twenty-four of 747 individuals demonstrated H5-specific T-cell responses but little or no cross-reactivity with H3 or H1 HA peptides. H5N1 peptide-specific T-cell lines that did not cross-react with H1 or H3 influenza virus HA peptides were generated. Four individuals also had antibodies against H5N1. This is the first report of ex vivo H5 HA-specific T-cell responses in a healthy but H5N1-exposed population. Our results indicate that the presence of H5N1-specific T cells could be an additional diagnostic tool for asymptomatic H5N1 infection.


The evolution of speed, size and shape in modern athletics.
In the present study, we show that the fastest runners and swimmers are becoming not only faster but also heavier, taller and more slender. During the past century, the world record speeds for 100 m-freestyle and 100 m-dash have increased with body mass (M) raised to the power 1/6, in accordance with the constructal scaling of animal locomotion. The world records also show that the speeds have increased in proportion with body heights (H) raised to the power 1/2, in accordance with animal locomotion scaling. If the athlete's body is modeled with two length scales (H, body width L), the (M, H) data can be used to calculate the slenderness of the body, H/L. The world records show that the body slenderness is increasing very slowly over time.


The extracellular RNA complement of Escherichia coli.
The secretion of biomolecules into the extracellular milieu is a common and well-conserved phenomenon in biology. In bacteria, secreted biomolecules are not only involved in intra-species communication but they also play roles in inter-kingdom exchanges and pathogenicity. To date, released products, such as small molecules, DNA, peptides, and proteins, have been well studied in bacteria. However, the bacterial extracellular RNA complement has so far not been comprehensively characterized. Here, we have analyzed, using a combination of physical characterization and high-throughput sequencing, the extracellular RNA complement of both outer membrane vesicle (OMV)-associated and OMV-free RNA of the enteric Gram-negative model bacterium Escherichia coli K-12 substrain MG1655 and have compared it to its intracellular RNA complement. Our results demonstrate that a large part of the extracellular RNA complement is in the size range between 15 and 40 nucleotides and is derived from specific intracellular RNAs. Furthermore, RNA is associated with OMVs and the relative abundances of RNA biotypes in the intracellular, OMV and OMV-free fractions are distinct. Apart from rRNA fragments, a significant portion of the extracellular RNA complement is composed of specific cleavage products of functionally important structural noncoding RNAs, including tRNAs, 4.5S RNA, 6S RNA, and tmRNA. In addition, the extracellular RNA pool includes RNA biotypes from cryptic prophages, intergenic, and coding regions, of which some are so far uncharacterised, for example, transcripts mapping to the fimA-fimL and ves-spy intergenic regions. Our study provides the first detailed characterization of the extracellular RNA complement of the enteric model bacterium E. coli. Analogous to findings in eukaryotes, our results suggest the selective export of specific RNA biotypes by E. coli, which in turn indicates a potential role for extracellular bacterial RNAs in intercellular communication.


Effects of acquired resistance on infection with Eimeria falciformis var. Pragensis in mice.
Mice immunized with infections of 500, 5,000, or 20,000 oocysts of E. falciformis var. pragensis were reinfected with 20,000 and 100,000 oocysts at 20 and 38 days, respectively, after the initial infection. After the first challenge infection, none of the immunized mice showed clinical signs of coccidiosis; a few mice passed very low numbers of oocysts, and oocyst discharge seemed to correlate negatively with immunizing dose. None of the mice immunized twice passed oocysts after challenge. Mice immunized with three infections were completely immune to challenge for 4 months. The effect of the immune response on the life cycle of the coccidium was determined by histological examination of the intestines of immune and nonimmune mice infected with the parasite. In both the immune and nonimmune groups, sporozoites penetrated absorptive epithelial cells and migrated to crypt epithelial cells during the first 6 to 24 h postinfection. At 48 to 72 h postinfection, the sporozoites developed into mature first-generation schizonts in the nonimmune mice, whereas the developing first-generation schizonts degenerated within the crypt epithelial cells of the immune mice. In nonimmune mice, third-generation merozoites, inoculated intracecally, developed into mature fourth-generation schizonts, whereas in immune mice the developing fourth-generation schizonts degenerated before maturing. The possibility that a cellmediated immune mechanism is responsible for the arrest in schizogony is discussed.


Basal rupture of saccular aneurysm. A pathological case report.
In a case of subarachnoid hemorrhage, the arteries of the circle of Willis were left undisturbed and undissected at postmortem examination. A block of frontal lobe tissue with the attached vessels was serially sectioned disclosing in its entirety a saccular aneurysm that had ruptured at the base rather than the dome. The histology of the hemostatic process was clearly depicted.


Genotype imputation reference panel selection using maximal phylogenetic diversity.
The recent dramatic cost reduction of next-generation sequencing technology enables investigators to assess most variants in the human genome to identify risk variants for complex diseases. However, sequencing large samples remains very expensive. For a study sample with existing genotype data, such as array data from genome-wide association studies, a cost-effective approach is to sequence a subset of the study sample and then to impute the rest of the study sample, using the sequenced subset as a reference panel. The use of such an internal reference panel identifies population-specific variants and avoids the problem of a substantial mismatch in ancestry background between the study population and the reference population. To efficiently select an internal panel, we introduce an idea of phylogenetic diversity from mathematical phylogenetics and comparative genomics. We propose the "most diverse reference panel", defined as the subset with the maximal "phylogenetic diversity", thereby incorporating individuals that span a diverse range of genotypes within the sample. Using data both from simulations and from the 1000 Genomes Project, we show that the most diverse reference panel can substantially improve the imputation accuracy compared to randomly selected reference panels, especially for the imputation of rare variants. The improvement in imputation accuracy holds across different marker densities, reference panel sizes, and lengths for the imputed segments. We thus propose a novel strategy for planning sequencing studies on samples with existing genotype data.


Double primary tumors-renal cell carcinoma and duodenal mucinous adenocarcinoma.
A 59-year old patient was admited to the Gastroenterology Clinic with the signs of gastrointestinal bleeding. Computerized tomography (CT) and a barium-meal radiography revealed a circumferential nodular wall narrowing and incomplete stricture at the D2 part of the duodenum. CT also showed a poorly demarcated mass in the upper and lower poles of the left kidney. During the operation, the whole kidney together with the tumor was removed and also a part of the duodenum. Morphological features of both tumors were typical and distinctive enough to set the diagnosis of two independent primary tumors. The possibility of one being the metastasis of the other was excluded. The diagnosis of double primary malignant neoplasms - renal cell carcinoma and duodenal mucinous adenocarcinoma was made.


Differential distribution of protein kinases along the crypt-to-lumen regions of rat colonic epithelium.
The activity of cAMP-dependent and cAMP-independent protein kinases, a class of enzymes involved in the regulation of cell proliferation was measured in rat colonic epithelium. Sequential cell populations harvested by a stepwise scraping technique from colonic crypt regions were identified by histology and incorporation of [3H]-thymidine into DNA. cAMP-independent phosphorylation of casein, in the presence of [gamma-32P]ATP, was markedly suppressed by quercetin, a bioflavonoid known to inhibit G-type casein kinase, protein kinase-C and tyrosine protein kinase. Conversely, the cyclic nucleotide regulatable form requiring histone as substrate was responsive to the action of the heat stable protein kinase inhibitor. The protein kinase species were characterised and partially purified by DEAE-cellulose chromatography. The activity of cAMP-dependent protein kinase in colonic cytosols (pmol 32P/min/mg protein, means (SE)) increased from 129.4 (15.9) in superficial cell populations to 238.5 (31.4) in lower crypt cell fractions (p less than 0.01). Colonic cAMP-independent protein kinase activity increased from 87.3 (15.6) in surface cell preparations to 178.1 (30.0) in lower crypt cell populations (p less than 0.02). A comparable activity gradient was observed in membrane fractions. The activity gradient persisted when the results were expressed as a function of cellular DNA. These findings indicate that protein kinases display a defined topological segregation along the colonic crypt regions and that during migration to the lumen colonic cells attenuate enzyme signals supposedly related to tissue growth.


Cumulative stress restricts niche filling potential of habitat-forming kelps in a future climate.
Climate change is driving range contractions and local population extinctions across the globe. When this affects ecosystem engineers the vacant niches left behind are likely to alter the wider ecosystem unless a similar species can fulfil them.Here, we explore the stress physiology of two coexisting kelps undergoing opposing range shifts in the Northeast Atlantic and discuss what differences in stress physiology may mean for future niche filling.We used chlorophyll florescence (Fv/Fm) and differentiation of the heat shock response (HSR) to determine the capacity of the expanding kelp, Laminaria ochroleuca, to move into the higher shore position of the retreating kelp, Laminaria digitata. We applied both single and consecutive exposures to immersed and emersed high and low temperature treatments, replicating low tide exposures experienced in summer and winter.No interspecific differences in HSR were observed which was surprising given the species' different biogeographic distributions. However, chlorophyll florescence revealed clear differences between species with L. ochroleuca better equipped to tolerate high immersed temperatures but showed little capacity to tolerate frosts or high emersion temperatures.Many patterns observed were only apparent after consecutive exposures. Such cumulative effects have largely been overlooked in tolerance experiments on intertidal organisms despite being more representative of the stress experienced in natural habitats. We therefore suggest future experiments incorporate consecutive stress into their design.Climate change is predicted to result in fewer ground frosts and increased summer temperatures. Therefore, L. ochroleuca may be released from its summer cold limit in winter but still be prevented from moving up the shore due to desiccation in the summer. Laminaria ochroleuca will, however, likely be able to move into tidal pools. Therefore, only partial niche filling by L. ochroleuca will be possible in this system as climate change advances. A plain language summary is available for this article.


An immunotoxin with therapeutic potential in T cell leukemia: WT1-ricin A.
A conjugate of the monoclonal antibody WT1 and ricin A-chain was studied for its suitability for purging marrow of leukemic T cells for autologous transplantation in T cell acute lymphocytic leukemia (T-ALL). The conjugate was powerfully cytotoxic to the human T-ALL cell line, GH1, which expresses the WT1 antigen at a high density. Treatment of the cells with the conjugate at 10(-11) M reduced their rate of protein synthesis by 50%, and the inclusion of 6 mM ammonium chloride in the cultures enhanced the potency of cytotoxic effect by 10-100-fold. Clonogenic assays indicated that less than 0.1% of GH1 cells survived 3-hr exposure to the conjugate in ammonium chloride. WT1 alone did not react with multipotent (CFU-GEMM) hematopoietic progenitors in normal human bone marrow, as measured by fluorescence-activated cell sorting. Under conditions giving maximal killing of GH1 cells, there was no toxicity to multipotential progenitors in normal human marrow.


Bipolar radio frequency endometrial ablation compared with balloon endometrial ablation in dysfunctional uterine bleeding: impact on patients' health-related quality of life.
To compare health-related quality of life (HRQoL) after bipolar radio frequency ablation and thermal balloon ablation in women with dysfunctional uterine bleeding. Randomized clinical trial. Teaching hospital. Women suffering from dysfunctional uterine bleeding. Bipolar radio frequency ablation and thermal balloon ablation. Patients were asked to complete HRQoL questionnaires at baseline, and at 2 days, 2 weeks, 3 months, 6 months, and 12 months after surgery. The questionnaires contained the medical outcomes study Short-Form 36 (SF-36), the Self-rating Depression Scale, the Rotterdam Symptom Checklist, State-Trait Anxiety Inventory, and a structured clinical history questionnaire. Data on HRQoL were available on at least two different time points in 115 of 126 randomized patients. HRQoL improved significantly over time in both groups, except for the domain of general health in the SF-36. None of the dimensions showed a significant difference between both groups, neither was there a significant interaction between time and treatment effect. Both methods significantly improved HRQoL in women with dysfunctional uterine bleeding. However, despite better amenorrhea and satisfaction rates after bipolar radio frequency ablation, there was no difference in HRQoL between the two groups.


Early detection of malignant pleural mesothelioma in asbestos-exposed individuals with a noninvasive proteomics-based surveillance tool.
Malignant pleural mesothelioma (MM) is an aggressive, asbestos-related pulmonary cancer that is increasing in incidence. Because diagnosis is difficult and the disease is relatively rare, most patients present at a clinically advanced stage where possibility of cure is minimal. To improve surveillance and detection of MM in the high-risk population, we completed a series of clinical studies to develop a noninvasive test for early detection. We conducted multi-center case-control studies in serum from 117 MM cases and 142 asbestos-exposed control individuals. Biomarker discovery, verification, and validation were performed using SOMAmer proteomic technology, which simultaneously measures over 1000 proteins in unfractionated biologic samples. Using univariate and multivariate approaches we discovered 64 candidate protein biomarkers and derived a 13-marker random forest classifier with an AUC of 0.99±0.01 in training, 0.98±0.04 in independent blinded verification and 0.95±0.04 in blinded validation studies. Sensitivity and specificity at our pre-specified decision threshold were 97%/92% in training and 90%/95% in blinded verification. This classifier accuracy was maintained in a second blinded validation set with a sensitivity/specificity of 90%/89% and combined accuracy of 92%. Sensitivity correlated with pathologic stage; 77% of Stage I, 93% of Stage II, 96% of Stage III and 96% of Stage IV cases were detected. An alternative decision threshold in the validation study yielding 98% specificity would still detect 60% of MM cases. In a paired sample set the classifier AUC of 0.99 and 91%/94% sensitivity/specificity was superior to that of mesothelin with an AUC of 0.82 and 66%/88% sensitivity/specificity. The candidate biomarker panel consists of both inflammatory and proliferative proteins, processes strongly associated with asbestos-induced malignancy. The SOMAmer biomarker panel discovered and validated in these studies provides a solid foundation for surveillance and diagnosis of MM in those at highest risk for this disease.


Optical characterization of photonic crystal slabs using orthogonally oriented polarization filters.
We present an experimental method for direct analysis of guided-mode resonances in photonic crystal slab structures using transmission measurements. By positioning the photonic crystal slab between orthogonally oriented polarization filters light transmission is suppressed except for the guided-mode resonances. Angle resolved transmission measurements with crossed polarizers are performed to obtain the band structure around the Gamma-point. Results are compared to mode simulations. Spatially resolved measurements in a confocal microscope setup are used for homogeneity characterizations. Stitching errors and inhomogeneities in exposure dose down to 1.3% in photonic crystal slabs fabricated by electron beam lithography are observed using this method.


Interstate migrants in the United States: some social-economic differences by type of move;.
U.S. interstate migrants (over a five-year period) are separated into three groups: (a) those leaving state of birth; (b) those returning to state of birth; and (c) those outside state of birth at the beginning of the period and moving on to a third state by the end. Evidence is presented suggesting that the third group is particularly selective of persons with high social and economic status. The findings are linked to certain hypotheses about the changing role and function of migration in a highly developed country where the transition from a rural society based on agriculture has essentially been completed.


Venous thromboembolism in women: a specific reproductive health risk.
BACKGROUND Venous thromboembolism (VTE) is a specific reproductive health risk for women. METHODS Searches were performed in Medline and other databases. The selection criteria were high-quality studies and studies relevant to clinical reproductive medicine. Summaries were presented and discussed by the European Society of Human Reproduction and Embryology Workshop Group. RESULTS VTE is a multifactorial disease with a baseline annual incidence around 50 per 100 000 at 25 years and 120 per 100 000 at age 50. Its major complication is pulmonary embolism, causing death in 1-2% of patients. Higher VTE risk is associated with an inherited thrombophilia in men and women. Changes in the coagulation system and in the risk of clinical VTE in women also occur during pregnancy, with the use of reproductive hormones and as a consequence of ovarian stimulation when hyperstimulation syndrome and conception occur together. In pregnancy, the risk of VTE is increased ~5-fold, while the use of combined hormonal contraception (CHC) doubles the risk and this relative risk is higher with the more recent pills containing desogestrel, gestodene and drospirenone when compared with those with levonorgestrel. Similarly, hormone replacement therapy (HRT) increases the VTE risk 2- to 4-fold. There is a synergistic effect between thrombophilia and the various reproductive risks. Prevention of VTE during pregnancy should be offered to women with specific risk factors. In women who are at high risk, CHC and HRT should be avoided. CONCLUSIONS Clinicians managing pregnancy or treating women for infertility or prescribing CHC and HRT should be aware of the increased risks of VTE and the need to take a careful medical history to identify additional co-existing risks, and should be able to diagnose VTE and know how to approach its prevention.


Exploring routine use of telemedicine through a case study in rehabilitation.
This study examines how telerehabilitation becomes part of existing and new clinical routines and identifies factors that enable or constrain its routine use. An in-depth case study of a telemedicine program in rehabilitation implemented between an urban specialized rehabilitation center and a rural regional rehabilitation center was conducted. Using a conceptual framework based on Giddens' theory of structuration, a qualitative analysis was carried out using four data sources: focus groups and phone interviews (with health professionals, managers, and patients and their family members); telerehabilitation video recordings; and project documents (e.g., proposals, requests for funding, summaries, agendas of meetings, operating procedures, patient handouts, and tools for clinicians). In two rehabilitation programs for 1) patients who sustained a traumatic brain injury and 2) those who sustained a spinal cord injury, telerehabilitation was successfully incorporated into routine clinical practices for activities such as interdisciplinary care plans. However, for specialized clinical consultations or long-term patient follow-up, telerehabilitation was not successfully incorporated. Factors that facilitated or prevented the integration of telerehabilitation in routine practices stemmed from both the structure (norms, rules, resources, and values) and the agent (e.g., users of telerehabilitation, including clinicians, managers, and patients and their families) and include 1) shared beliefs and assumptions held by patient care team members regarding the nature of the clinical activities, and the perceptions of patients and their family members; 2) clinical and organizational leadership; 3) extent and type of telerehabilitation use; 4) available resources; and 5) collaborations already in place or needing to be developed. This study provides empirical evidence of how telerehabilitation activities may become integrated into routine day-to-day clinical activities.


The structures of a naturally empty cowpea mosaic virus particle and its genome-containing counterpart by cryo-electron microscopy.
Cowpea mosaic virus (CPMV) is a picorna-like plant virus. As well as an intrinsic interest in CPMV as a plant pathogen, CPMV is of major interest in biotechnology applications such as nanotechnology. Here, we report high resolution cryo electron microscopy (cryo-EM) maps of wild type CPMV containing RNA-2, and of naturally-formed empty CPMV capsids. The resolution of these structures is sufficient to visualise large amino acids. We have refined an atomic model for each map and identified an essential amino acid involved in genome encapsidation. This work has furthered our knowledge of Picornavirales genome encapsidation and will assist further work in the development of CPMV as a biotechnological tool.


Lessons from genetic profiling in soft tissue sarcomas.
Soft tissue sarcomas represent a heterogeneous group of tumors and include over 50 histotypes. Some of these tumor types are characterized by specific chromosomal translocations, whereas other types show complex genetic aberrations. The recent developments within gene expression technologies have now been applied to studies of soft tissue sarcomas (STS) and the first results indicate that genetic signatures are useful for classification and diagnosis. Distinctive expression profiles have been found in e.g. gastrointestinal stromal tumors (GISTs), synovial sarcomas, malignant peripheral nerve sheath tumors (MPNSTs), and in subsets of liposarcomas. The more pleomorphic tumor types, such as high-grade variants of leiomyosarcomas, malignant fibrous histiocytomas (MFHs), fibrosarcomas, and subtypes of liposarcomas, show a greater variability among the expression profiles, but interestingly subsets with distinctive expression profiles can be identified also among these tumors. The data available place many of the genes hypothesized to be involved in the development of a certain type of STS, such as the KIT gene in GIST development, among the top discriminating genes. Thereby expression profiling provides novel insights into the pathogenesis of STS. Although much work remains to be done to validate the data and to define optimal discriminating gene lists, the current lessons from gene expression studies in STS are encouraging and imply that genetic signatures may serve as diagnostic and prognostic markers and may help identify novel therapeutic strategies.


Hyperhomocysteinemia disrupts retinal pigment epithelial structure and function with features of age-related macular degeneration.
The disruption of retinal pigment epithelial (RPE) function and the degeneration of photoreceptors are cardinal features of age related macular degeneration (AMD); however there are still gaps in our understanding of underlying biological processes. Excess homocysteine (Hcy) has been reported to be elevated in plasma of patients with AMD. This study aimed to evaluate the direct effect of hyperhomocysteinemia (HHcy) on structure and function of RPE. Initial studies in a mouse model of HHcy, in which cystathionine-β-synthase (cbs) was deficient, revealed abnormal RPE cell morphology with features similar to that of AMD upon optical coherence tomography (OCT), fluorescein angiography (FA), histological, and electron microscopic examinations. These features include atrophy, vacuolization, hypopigmentation, thickened basal laminar membrane, hyporeflective lucency, choroidal neovascularization (CNV), and disturbed RPE-photoreceptor relationship. Furthermore, intravitreal injection of Hcy per se in normal wild type (WT) mice resulted in diffuse hyper-fluorescence, albumin leakage, and CNV in the area of RPE. In vitro experiments on ARPE-19 showed that Hcy dose-dependently reduced tight junction protein expression, increased FITC dextran leakage, decreased transcellular electrical resistance, and impaired phagocytic activity. Collectively, our results demonstrated unreported effects of excess Hcy levels on RPE structure and function that lead to the development of AMD-like features.


Modeling HIV-1 drug resistance as episodic directional selection.
The evolution of substitutions conferring drug resistance to HIV-1 is both episodic, occurring when patients are on antiretroviral therapy, and strongly directional, with site-specific resistant residues increasing in frequency over time. While methods exist to detect episodic diversifying selection and continuous directional selection, no evolutionary model combining these two properties has been proposed. We present two models of episodic directional selection (MEDS and EDEPS) which allow the a priori specification of lineages expected to have undergone directional selection. The models infer the sites and target residues that were likely subject to directional selection, using either codon or protein sequences. Compared to its null model of episodic diversifying selection, MEDS provides a superior fit to most sites known to be involved in drug resistance, and neither one test for episodic diversifying selection nor another for constant directional selection are able to detect as many true positives as MEDS and EDEPS while maintaining acceptable levels of false positives. This suggests that episodic directional selection is a better description of the process driving the evolution of drug resistance.


First Report of Dollar Spot Caused by Sclerotinia homoeocarpa on Switchgrass in the United States.
Sclerotinia homoeocarpa causes dollar spot on many grass species; however, it has not been described on switchgrass (Panicum virgatum L.) as a host. In August 2010, bleached, tan-to-straw-colored leaf spots with dark brown-to-reddish brown margins were found in patchy distribution in small field plots of 'Alamo' switchgrass at the East Tennessee Research and Education Center, Knoxville, TN. The plots had been planted to switchgrass for the past 21 years. Disease lesions covered 75 to 80% of leaf tissue per patch and were also evident on stems. To identify the pathogen, center portions of diseased leaves were cut into 20- to 30-cm segments, surface disinfested (95% ethanol for 30 s, 10% bleach for 1 min, and 95% ethanol for 30 s), and dried. Disinfested leaves (5-cm sections that included a leading edge of a lesion) were plated on potato dextrose agar (PDA). Plates were incubated at 22°C. Within 12 h, white, fluffy, aerial mycelium developed. Viewed from above, colonies were tan to cinnamon in color with a dark brown-to-black substratal stroma on and in the agar, which appeared brown as viewed from below the petri dish. No spores were observed. Morphological characteristics of colony and hyphal growth were identical to those of S. homoeocarpa F.T. Bennett (1). Pathogenicity studies were conducted with 6-week-old 'Alamo' switchgrass grown from scarified (2), surface-disinfested seed. Nine (9 × 9-cm2) pots with 18 plants each were inoculated with 20 mycelial plugs (6-mm diameter) per pot, taken from 3-to-5-day-old fungal cultures. Two control pots were inoculated with sterile PDA plugs and subjected to the same conditions. Plugs were placed on leaf surfaces and around the plant crowns. Plants were subjected to high humidity by enclosure in a plastic bag and incubated in a growth chamber at 25/20°C with a 12-h photoperiod. Plastic bags were removed after 48 h. Leaf spots appeared as early as 2 days postinoculation, with full symptoms after 2 weeks for eight of nine replicates. Control plants had no symptoms. The fungus was cultured from leaf spots and stem lesions of inoculated plants as described above. The same disease and fungus were observed, completing Koch's postulates. The internal transcribed spacer (ITS) regions of ribosomal DNA from the original isolate used for inoculation and from the isolate recovered from plants in the pathogenicity assay were amplified with PCR with primers ITS4 and ITS5 (4). PCR amplicons of ~565 bp were sequenced; sequences of amplicons from the original isolate and reisolate were identical and submitted to GenBank (Accession No. HQ850151). The sequence had 99% homology with several S. homoeocarpa isolates in GenBank, including three isolates from buffalograss in Oklahoma (Accession Nos. EU123800, EU123802, and EU123803). The mitochondrial small subunit region was amplified from the original isolate with primers NMS1 and NMS2 (3). The resultant 536-bp fragment was sequenced and submitted to GenBank (Accession No. HQ850152), but no S. homoeocarpa sequences were available for comparison. To our knowledge, this is the first confirmed report of switchgrass as a natural host for S. homoeocarpa, extending the known host range for the pathogen. References: (1) F. T. Bennett. Ann. Appl. Biol. 24:236, 1937. (2) K. D. Gwinn et al. Crop Sci. 31:1369, 1991. (3) K. N. Li et al. Appl. Environ. Microbiol. 60:4324, 1994. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, NY, 1990.


Face mask use during the COVID-19 pandemic: how risk perception, experience with COVID-19, and attitude towards government interact with country-wide policy stringency.
During the 2020 COVID-19 pandemic, governments imposed numerous regulations to protect public health, particularly the (mandatory) use of face masks. However, the appropriateness and effectiveness of face mask regulations have been widely discussed, as is apparent from the divergent measures taken across and within countries over time, including mandating, recommending, and discouraging their use. In this study, we analyse how country-level policy stringency and individual-level predictors associate with face mask use during the early stages of the global COVID-19 pandemic. First, we study how (self and other-related) risk perception, (direct and indirect) experience with COVID-19, attitude towards government and policy stringency shape face mask use. Second, we study whether there is an interaction between policy stringency and the individual-level variables. We conduct multilevel analyses exploiting variation in face mask regulations across countries and using data from approximately 7000 students collected in the beginning of the pandemic (weeks 17 through 19, 2020). We show that policy stringency is strongly positively associated with face mask use. We find a positive association between self-related risk perception and mask use, but no relationship of mask use with experience with COVID-19 and attitudes towards government. However, in the interaction analyses, we find that government trust and perceived clarity of communication moderate the link between stringency and mask use, with positive government perceptions relating to higher use in countries with regulations and to lower use in countries without regulations. We highlight that those countries that aim for widespread use of face masks should set strict measures, stress self-related risks of COVID-19, and use clear communication.


Acute airway management.
Acute airway management has challenged clinicians for nearly four millennia. History underscores the discoveries of surgeons and anesthesiologists, whose advances in technology and surgical technique have transformed management algorithms from primarily surgical tracheostomy, to transoral endotracheal intubation under direct laryngoscopy. Despite this progress and a better understanding of airway anatomy, physiology and pathogenesis of disease, the acute airway, whether obstructed, traumatically disrupted, or externally compressed, remains a life-threatening challenge. The role of all clinicians in acute airway management is patient stability and emergent control of the airway to ensure patency as well as adequate oxygenation and ventilation. The standard of care remains transoral intubation under direct laryngoscopy with use of indirect laryngoscopy as a first adjunct. If unsuccessful, surgical intubation of the trachea via cricothyroidotomy with subsequent conversion to open tracheostomy is the procedure of choice. While there is growing support for the use of percutaneous tracheostomy as an alternative to surgical intubation of the trachea after failed transoral intubation, the potential for damage to critical neck structures and longer time-to-intubation must be considered. In this perspective, we provide a history of acute airway management, highlighting milestones in the fields of airway surgery and anesthesia. We present a review of current medical and surgical approaches to managing the acute airway, including the risks, benefits and appropriateness of each approach with respect to patient stability, available equipment, clinician training and patient outcomes. We conclude with an emphasis on the role of the thoracic surgeon in prevention and the critical nature of regular surveillance of patients with chronic, partial tracheal obstruction.