A wirelessly powered and controlled device for optical neural control of freely-behaving animals.
Optogenetics, the ability to use light to activate and silence specific neuron types within neural networks in vivo and in vitro, is revolutionizing neuroscientists' capacity to understand how defined neural circuit elements contribute to normal and pathological brain functions. Typically, awake behaving experiments are conducted by inserting an optical fiber into the brain, tethered to a remote laser, or by utilizing an implanted light-emitting diode (LED), tethered to a remote power source. A fully wireless system would enable chronic or longitudinal experiments where long duration tethering is impractical, and would also support high-throughput experimentation. However, the high power requirements of light sources (LEDs, lasers), especially in the context of the extended illumination periods often desired in experiments, precludes battery-powered approaches from being widely applicable. We have developed a headborne device weighing 2 g capable of wirelessly receiving power using a resonant RF power link and storing the energy in an adaptive supercapacitor circuit, which can algorithmically control one or more headborne LEDs via a microcontroller. The device can deliver approximately 2 W of power to the LEDs in steady state, and 4.3 W in bursts. We also present an optional radio transceiver module (1 g) which, when added to the base headborne device, enables real-time updating of light delivery protocols; dozens of devices can be controlled simultaneously from one computer. We demonstrate use of the technology to wirelessly drive cortical control of movement in mice. These devices may serve as prototypes for clinical ultra-precise neural prosthetics that use light as the modality of biological control.


Condensation and dissociation rates for gas phase metal clusters from molecular dynamics trajectory calculations.
In gas phase synthesis systems, clusters form and grow via condensation, in which a monomer binds to an existing cluster. While a hard-sphere equation is frequently used to predict the condensation rate coefficient, this equation neglects the influences of potential interactions and cluster internal energy on the condensation process. Here, we present a collision rate theory-molecular dynamics simulation approach to calculate condensation probabilities and condensation rate coefficients. We use this approach to examine atomic condensation onto 6-56-atom Au and Mg clusters. The probability of condensation depends upon the initial relative velocity (v) between atom and cluster and the initial impact parameter (b). In all cases, there is a well-defined region of b-v space where condensation is highly probable, and outside of which the condensation probability drops to zero. For Au clusters with more than 10 atoms, we find that at gas temperatures in the 300-1200 K range, the condensation rate coefficient exceeds the hard-sphere rate coefficient by a factor of 1.5-2.0. Conversely, for Au clusters with 10 or fewer atoms and for 14- and 28-atom Mg clusters, as cluster equilibration temperature increases, the condensation rate coefficient drops to values below the hard-sphere rate coefficient. Calculations also yield the self-dissociation rate coefficient, which is found to vary considerably with gas temperature. Finally, calculations results reveal that grazing (high b) atom-cluster collisions at elevated velocity (>1000 m s-1) can result in the colliding atom rebounding (bounce) from the cluster surface or binding while another atom dissociates (replacement). The presented method can be applied in developing rate equations to predict material formation and growth rates in vapor phase systems.


Ecological relationships of meso-scale distribution in 25 neotropical vertebrate species.
Vertebrates are a vital ecological component of Amazon forest biodiversity. Although vertebrates are a functionally important part of various ecosystem services they continue to be threatened by anthropogenic impacts throughout the Amazon. Here we use a standardized, regularly spaced arrangement of camera traps within 25km2 to provide a baseline assessment of vertebrate species diversity in a sustainable use protected area in the eastern Brazilian Amazon. We examined seasonal differences in the per species encounter rates (number of photos per camera trap and number of cameras with photos). Generalized linear models (GLMs) were then used to examine the influence of five variables (altitude, canopy cover, basal area, distance to nearest river and distance to nearest large river) on the number of photos per species and on functional groups. GLMs were also used to examine the relationships between large predators [Jaguar (Panthera onca) and Puma (Puma concolor)] and their prey. A total of 649 independent photos of 25 species were obtained from 1,800 camera trap days (900 each during wet and dry seasons). Only ungulates and rodents showed significant seasonal differences in the number of photos per camera. The number of photos differed between seasons for only three species (Mazama americana, Dasyprocta leporina and Myoprocta acouchy) all of which were photographed more (3 to 10 fold increase) during the wet season. Mazama americana was the only species where a significant difference was found in occupancy, with more photos in more cameras during the wet season. For most groups and species variation in the number of photos per camera was only explained weakly by the GLMs (deviance explained ranging from 10.3 to 54.4%). Terrestrial birds (Crax alector, Psophia crepitans and Tinamus major) and rodents (Cuniculus paca, Dasyprocta leporina and M. acouchy) were the notable exceptions, with our GLMs significantly explaining variation in the distribution of all species (deviance explained ranging from 21.0 to 54.5%). The group and species GLMs showed some novel ecological information from this relatively pristine area. We found no association between large cats and their potential prey. We also found that rodent and bird species were more often recorded closer to streams. As hunters gain access via rivers this finding suggests that there is currently little anthropogenic impact on the species. Our findings provide a standardized baseline for comparison with other sites and with which planned management and extractive activities can be evaluated.


New predictors of complications in carotid body tumor resection.
This study examined the relationship between two new variables, tumor distance to base of skull (DTBOS) and tumor volume, with complications of carotid body tumor (CBT) resection, including bleeding and cranial nerve injury. Patients who underwent CBT resection between 2004 and 2014 were studied using a standardized, multi-institutional database. Demographic, perioperative, and outcomes data were collected. CBT measurements were determined from computed tomography, magnetic resonance imaging, and ultrasound examination. There were 356 CBTs resected in 332 patients (mean age, 51 years; 72% female); 32% were classified as Shamblin I, 43% as Shamblin II, and 23% as Shamblin III. The mean DTBOS was 3.3 cm (standard deviation [SD], 2.1; range, 0-10), and the mean tumor volume was 209.7 cm3 (SD, 266.7; range, 1.1-1642.0 cm3). The mean estimated blood loss (EBL) was 257 mL (SD, 426; range, 0-3500 mL). Twenty-four percent of patients had cranial nerve injuries. The most common cranial nerves injured were the hypoglossal (10%), vagus (11%), and superior laryngeal (5%) nerves. Both Shamblin grade and DTBOS were statistically significantly correlated with EBL of surgery and cranial nerve injuries, whereas tumor volume was statistically significantly correlated with EBL. The logistic model for predicting blood loss and cranial nerve injury with all three variables-Shamblin, DTBOS, and volume (R2 = 0.171, 0.221, respectively)-was superior to a model with Shamblin alone (R2 = 0.043, 0.091, respectively). After adjusting for Shamblin grade and volume, every 1-cm decrease in DTBOS was associated with 1.8 times increase in risk of >250 mL of blood loss (95% confidence interval, 1.25-2.55) and 1.5 times increased risk of cranial nerve injury (95% confidence interval, 1.19-1.92). This large study of CBTs demonstrates the value of preoperatively determining tumor dimensions and how far the tumor is located from the base of the skull. DTBOS and tumor volume, when used in combination with the Shamblin grade, better predict bleeding and cranial nerve injury risk. Furthermore, surgical resection before expansion toward the base of the skull reduces complications as every 1-cm decrease in the distance to the skull base results in 1.8 times increase in >250 mL of blood loss and 1.5 times increased risk of cranial nerve injury.


Palliative care provision for patients with chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) is a major cause of disability, morbidity and mortality in old age. Patients with advanced stage COPD are most likely to be admitted three to four times per year with acute exacerbations of COPD (AECOPD) which are costly to manage. The adverse events of AECOPD are associated with poor quality of life, severe physical disability, loneliness, and depression and anxiety symptoms. Currently there is a lack of palliative care provision for patients with advanced stage COPD compared with cancer patients despite having poor prognosis, intolerable dyspnoea, lower levels of self efficacy, greater disability, poor quality of life and higher levels of anxiety and depression. These symptoms affect patients' quality of life and can be a source of concern for family and carers as most patients are likely to be housebound and may be in need of continuous support and care. Evidence of palliative care provision for cancer patients indicate that it improves quality of life and reduces health care costs. The reasons why COPD patients do not receive palliative care are complex. This partly may relate to prognostic accuracy of patients' survival which poses a challenge for healthcare professionals, including general practitioners for patients with advanced stage COPD, as they are less likely to engage in end-of-life care planning in contrast with terminal disease like cancer. Furthermore there is a lack of resources which constraints for the wider availability of the palliative care programmes in the health care system. Potential barriers may include unwillingness of patients to discuss advance care planning and end-of-life care with their general practitioners, lack of time, increased workload, and fear of uncertainty of the information to provide about the prognosis of the disease and also lack of appropriate tools to guide general practitioners when to refer patients for palliative care. COPD is a chronic incurable disease; those in an advanced stage of the disease pursuing intensive medical treatment may also benefit from the simultaneous holistic care approach of palliative care services, medical services and social services to improve quality of end of life care.


Studying gene and gene-environment effects of uncommon and common variants on continuous traits: a marker-set approach using gene-trait similarity regression.
Genomic association analyses of complex traits demand statistical tools that are capable of detecting small effects of common and rare variants and modeling complex interaction effects and yet are computationally feasible. In this work, we introduce a similarity-based regression method for assessing the main genetic and interaction effects of a group of markers on quantitative traits. The method uses genetic similarity to aggregate information from multiple polymorphic sites and integrates adaptive weights that depend on allele frequencies to accomodate common and uncommon variants. Collapsing information at the similarity level instead of the genotype level avoids canceling signals that have the opposite etiological effects and is applicable to any class of genetic variants without the need for dichotomizing the allele types. To assess gene-trait associations, we regress trait similarities for pairs of unrelated individuals on their genetic similarities and assess association by using a score test whose limiting distribution is derived in this work. The proposed regression framework allows for covariates, has the capacity to model both main and interaction effects, can be applied to a mixture of different polymorphism types, and is computationally efficient. These features make it an ideal tool for evaluating associations between phenotype and marker sets defined by linkage disequilibrium (LD) blocks, genes, or pathways in whole-genome analysis.


Characterization of the collagen-binding S-layer protein CbsA of Lactobacillus crispatus.
The cbsA gene of Lactobacillus crispatus strain JCM 5810, encoding a protein that mediates adhesiveness to collagens, was characterized and expressed in Escherichia coli. The cbsA open reading frame encoded a signal sequence of 30 amino acids and a mature polypeptide of 410 amino acids with typical features of a bacterial S-layer protein. The cbsA gene product was expressed as a His tag fusion protein, purified by affinity chromatography, and shown to bind solubilized as well as immobilized type I and IV collagens. Three other Lactobacillus S-layer proteins, SlpA, CbsB, and SlpnB, bound collagens only weakly, and sequence comparisons of CbsA with these S-layer proteins were used to select sites in cbsA where deletions and mutations were introduced. In addition, hybrid S-layer proteins that contained the N or the C terminus from CbsA, SlpA, or SlpnB as well as N- and C-terminally truncated peptides from CbsA were constructed by gene fusion. Analysis of these molecules revealed the major collagen-binding region within the N-terminal 287 residues and a weaker type I collagen-binding region in the C terminus of the CbsA molecule. The mutated or hybrid CbsA molecules and peptides that failed to polymerize into a periodic S-layer did not bind collagens, suggesting that the crystal structure with a regular array is optimal for expression of collagen binding by CbsA. Strain JCM 5810 was found to contain another S-layer gene termed cbsB that was 44% identical in sequence to cbsA. RNA analysis showed that cbsA, but not cbsB, was transcribed under laboratory conditions. S-layer-protein-expressing cells of strain JCM 5810 adhered to collagen-containing regions in the chicken colon, suggesting that CbsA-mediated collagen binding represents a true tissue adherence property of L. crispatus.


Integration of microwave tomography with magnetic resonance for improved breast imaging.
Breast magnetic resonance imaging is highly sensitive but not very specific for the detection of breast cancer. Opportunities exist to supplement the image acquisition with a more specific modality provided the technical challenges of meeting space limitations inside the bore, restricted breast access, and electromagnetic compatibility requirements can be overcome. Magnetic resonance (MR) and microwave tomography (MT) are complementary and synergistic because the high resolution of MR is used to encode spatial priors on breast geometry and internal parenchymal features that have distinct electrical properties (i.e., fat vs fibroglandular tissue) for microwave tomography. The authors have overcome integration challenges associated with combining MT with MR to produce a new coregistered, multimodality breast imaging platform--magnetic resonance microwave tomography, including: substantial illumination tank size reduction specific to the confined MR bore diameter, minimization of metal content and composition, reduction of metal artifacts in the MR images, and suppression of unwanted MT multipath signals. MR SNR exceeding 40 dB can be obtained. Proper filtering of MR signals reduces MT data degradation allowing MT SNR of 20 dB to be obtained, which is sufficient for image reconstruction. When MR spatial priors are incorporated into the recovery of MT property estimates, the errors between the recovered versus actual dielectric properties approach 5%. The phantom and human subject exams presented here are the first demonstration of combining MT with MR to improve the accuracy of the reconstructed MT images.


Effect of background components on spatial-frequency masking.
Previous studies of spatial-frequency masking and adaptation have shown that the contrast-detection threshold elevates maximally when the test spatial frequency is the same as the masking (or adapting) frequency but changes only slightly when they are separated by two or more octaves. At low spatial frequencies, however, the peak of the threshold-elevation function does not obey this rule: there is a well-established peak shift in the threshold-elevation functions toward higher spatial frequencies. We investigated whether this shift might be due to the masking effects caused by the background field, which contributes energy at the very low end of the spectrum. We first measured the effect of a 3-cycles/deg (c/deg) mask on detection of a range of test frequencies, compared with unmasked detection thresholds. We then measured the combined effect of a 2-c/deg and a 3-c/deg mask on detection, compared with detection with just the 2-c/deg mask. The comparison in the second case still tests the effect of the 3-c/deg mask, but the presence of the hidden 2-c/deg mask causes the peak masking effect to shift toward higher frequencies. This result provides a proof of concept for the hypothesis that the peak shift at low spatial frequencies is caused by the low-frequency energy in the background field, which is present in both masked and unmasked conditions. A five-parameter quantitative model of frequency masking is presented that describes the pure contrast-detection function, the frequency-masking functions at mask frequencies of 0.25, 0.5, 2, and 3 c/deg, and the peak-shift phenomenon.


Carboxypeptidase A6 in zebrafish development and implications for VIth cranial nerve pathfinding.
Carboxypeptidase A6 (CPA6) is an extracellular protease that cleaves carboxy-terminal hydrophobic amino acids and has been implicated in the defective innervation of the lateral rectus muscle by the VIth cranial nerve in Duane syndrome. In order to investigate the role of CPA6 in development, in particular its potential role in axon guidance, the zebrafish ortholog was identified and cloned. Zebrafish CPA6 was secreted and interacted with the extracellular matrix where it had a neutral pH optimum and specificity for C-terminal hydrophobic amino acids. Transient mRNA expression was found in newly formed somites, pectoral fin buds, the stomodeum and a conspicuous condensation posterior to the eye. Markers showed this tissue was not myogenic in nature. Rather, the CPA6 localization overlapped with a chondrogenic site which subsequently forms the walls of a myodome surrounding the lateral rectus muscle. No other zebrafish CPA gene exhibited a similar expression profile. Morpholino-mediated knockdown of CPA6 combined with retrograde labeling and horizontal eye movement analyses demonstrated that deficiency of CPA6 alone did not affect either VIth nerve development or function in the zebrafish. We suggest that mutations in other genes and/or enhancer elements, together with defective CPA6 expression, may be required for altered VIth nerve pathfinding. If mutations in CPA6 contribute to Duane syndrome, our results also suggest that Duane syndrome can be a chondrogenic rather than a myogenic or neurogenic developmental disorder.


15-Lipoxygenase-2 expression in benign and neoplastic lung: an immunohistochemical study and correlation with tumor grade and proliferation.
15-Lipoxygenase-2 (15-LOX-2) is an arachidonic acid-metabolizing enzyme expressed in prostate, lung, skin, esophagus, and cornea. In the benign prostate, it is expressed in differentiated secretory epithelial cells, where its enzymatic product 15-HETE may regulate transcription by activating the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma). 15-LOX-2 and 15-HETE formation are reduced in prostate carcinoma. The distribution of 15-LOX-2 in the normal lung and its expression in lung carcinomas has not been reported and was investigated in the current study by using immunohistochemistry and tissue microarrays (TMAs). In benign lung, 15-LOX-2 immunostaining was noted exclusively in type II pneumocytes, which are known to express PPARgamma. Of 160 lung carcinomas, 15-LOX-2 was expressed in non-small cell carcinomas (NSCLC), including 33 of 69 (48%) adenocarcinomas, with 10 of 16 (63%) bronchioloalveolar carcinomas immunopositive. Fourteen of 55 (25%) squamous cell carcinomas and 2 of 14 (14%) large cell carcinomas showed weak immunostaining. All 19 neuroendocrine tumors were negative. Better differentiated NSCLCs showed greater 15-LOX-2 expression, with a significant inverse correlation between 15-LOX-2 immunostaining and tumor grade (P < 0.03). A significant inverse correlation was also noted between 15-LOX-2 immunostaining and tumor cell proliferation (Ki-67 immunostaining; P < 0.0001). These findings suggest a possible role of 15-LOX-2 in regulating secretory differentiation and proliferation in benign lung and NSCLCs, particularly adenocarcinomas.


Flaviviruses are sensitive to inhibition of thymidine synthesis pathways.
Dengue virus has emerged as a global health threat to over one-third of humankind. As a positive-strand RNA virus, dengue virus relies on the host cell metabolism for its translation, replication, and egress. Therefore, a better understanding of the host cell metabolic pathways required for dengue virus infection offers the opportunity to develop new approaches for therapeutic intervention. In a recently described screen of known drugs and bioactive molecules, we observed that methotrexate and floxuridine inhibited dengue virus infections at low micromolar concentrations. Here, we demonstrate that all serotypes of dengue virus, as well as West Nile virus, are highly sensitive to both methotrexate and floxuridine, whereas other RNA viruses (Sindbis virus and vesicular stomatitis virus) are not. Interestingly, flavivirus replication was restored by folinic acid, a thymidine precursor, in the presence of methotrexate and by thymidine in the presence of floxuridine, suggesting an unexpected role for thymidine in flavivirus replication. Since thymidine is not incorporated into RNA genomes, it is likely that increased thymidine production is indirectly involved in flavivirus replication. A possible mechanism is suggested by the finding that p53 inhibition restored dengue virus replication in the presence of floxuridine, consistent with thymidine-less stress triggering p53-mediated antiflavivirus effects in infected cells. Our data reveal thymidine synthesis pathways as new and unexpected therapeutic targets for antiflaviviral drug development.


Asthma progression and mortality: the role of inhaled corticosteroids.
Overall, asthma mortality rates have declined dramatically in the last 30 years, due to improved diagnosis and to better treatment, particularly in the 1990s following the more widespread use of inhaled corticosteroids (ICSs). The impact of ICS on other long-term outcomes, such as lung function decline, is less certain, in part because the factors associated with these outcomes are incompletely understood. The purpose of this review is to evaluate the effect of pharmacological interventions, particularly ICS, on asthma progression and mortality. Furthermore, we review the potential mechanisms of action of pharmacotherapy on asthma progression and mortality, the effects of ICS on long-term changes in lung function, and the role of ICS in various asthma phenotypes.Overall, there is compelling evidence of the value of ICS in improving asthma control, as measured by improved symptoms, pulmonary function and reduced exacerbations. There is, however, less convincing evidence that ICS prevents the decline in pulmonary function that occurs in some, although not all, patients with asthma. Severe exacerbations are associated with a more rapid decline in pulmonary function, and by reducing the risk of severe exacerbations, it is likely that ICS will, at least partially, prevent this decline. Studies using administrative databases also support an important role for ICS in reducing asthma mortality, but the fact that asthma mortality is, fortunately, an uncommon event makes it highly improbable that this will be demonstrated in prospective trials.


A Fatal Twist: Volvulus of the Small Intestine in a 46-Year-Old Woman.
A 46-year-old woman presented to two emergency departments within 12 hours because of acute abdominal pain. Physical exam demonstrated tenderness and epigastric guarding. An ultrasound was interpreted as negative; she was discharged home. Later that evening, she was found dead. Postmortem exam revealed acute hemorrhagic necrosis of a segment of jejunum secondary to volvulus. Clinical clues suggesting presentations of small bowel volvulus are usually nonspecific; the diagnosis is typically confirmed at surgery. Her unremitting abdominal pain, persistent vomiting, and absolute neutrophilia were consistent with an acute process. The etiology of this volvulus was caused by an elastic fibrous band at the root of the jejunal mesentery. While congenital fibrous bands are rare in adults, this interpretation is favored for two reasons. First, the band was located 20 cm superior to postsurgical adhesions in the lower abdomen and pelvis. Second, there was no history of trauma or previous surgery involving the site of volvulus.


Sexual dimorphism of male face shape, partnership status and the temporal context of relationship sought modulate women's preferences for direct gaze.
Most previous studies of face preferences have investigated the physical cues that influence face preferences. Far fewer studies have investigated the effects of cues to the direction of others' social interest (i.e. gaze direction) on face preferences. Here we found that unpartnered women demonstrated stronger preferences for direct gaze (indicating social interest) from feminine male faces than from masculine male faces when judging men's attractiveness for long-term relationships, but not when judging men's attractiveness for short-term relationships. Moreover, unpartnered women's preferences for direct gaze from feminine men were stronger for long-term than short-term relationships, but there was no comparable effect for judgements of masculine men. No such effects were evident among women with romantic partners, potentially reflecting different motivations underlying partnered and unpartnered women's judgements of men's attractiveness. Collectively these findings (1) complement previous findings whereby women demonstrated stronger preferences for feminine men as long-term than short-term partners, (2) demonstrate context-sensitivity in the integration of physical and social cues in face preferences, and (3) suggest that gaze preferences may function, at least in part, to facilitate efficient allocation of mating effort.


Current status of antifungal susceptibility testing methods.
Antifungal susceptibility testing is a very dynamic field of medical mycology. Standardization of in vitro susceptibility tests by the Clinical and Laboratory Standards Institute (CLSI) and the European Committee for Antimicrobial Susceptibility Testing (EUCAST), and current availability of reference methods constituted the major remarkable steps in the field. Based on the established minimum inhibitory concentration (MIC) breakpoints, it is now possible to determine the susceptibilities of Candida strains to fluconazole, itraconazole, voriconazole, and flucytosine. Moreover, utility of fluconazole antifungal susceptibility tests as an adjunct in optimizing treatment of candidiasis has now been validated. While the MIC breakpoints and clinical significance of susceptibility testing for the remaining fungi and antifungal drugs remain yet unclear, modifications of the available methods as well as other methodologies are being intensively studied to overcome the present drawbacks and limitations. Among the other methods under investigation are Etest, colorimetric microdilution, agar dilution, determination of fungicidal activity, flow cytometry, and ergosterol quantitation. Etest offers the advantage of practical application and favorable agreement rates with the reference methods that are frequently above acceptable limits. However, MIC breakpoints for Etest remain to be evaluated and established. Development of commercially available, standardized colorimetric panels that are based on CLSI method parameters has added more to the antifungal susceptibility testing armamentarium. Flow cytometry, on the other hand, appears to offer rapid susceptibility testing but requires specified equipment and further evaluation for reproducibility and standardization. Ergosterol quantitation is another novel approach, which appears potentially beneficial particularly in discrimination of azole-resistant isolates from heavy trailers. The method is yet investigational and requires to be further studied. Developments in methodology and applications of antifungal susceptibility testing will hopefully provide enhanced utility in clinical guidance of antifungal therapy. However, and particularly in immunosuppressed host, in vitro susceptibility is and will remain only one of several factors that influence clinical outcome.


The burden of disease in older people and implications for health policy and practice.
23% of the total global burden of disease is attributable to disorders in people aged 60 years and older. Although the proportion of the burden arising from older people (≥60 years) is highest in high-income regions, disability-adjusted life years (DALYs) per head are 40% higher in low-income and middle-income regions, accounted for by the increased burden per head of population arising from cardiovascular diseases, and sensory, respiratory, and infectious disorders. The leading contributors to disease burden in older people are cardiovascular diseases (30·3% of the total burden in people aged 60 years and older), malignant neoplasms (15·1%), chronic respiratory diseases (9·5%), musculoskeletal diseases (7·5%), and neurological and mental disorders (6·6%). A substantial and increased proportion of morbidity and mortality due to chronic disease occurs in older people. Primary prevention in adults aged younger than 60 years will improve health in successive cohorts of older people, but much of the potential to reduce disease burden will come from more effective primary, secondary, and tertiary prevention targeting older people. Obstacles include misplaced global health priorities, ageism, the poor preparedness of health systems to deliver age-appropriate care for chronic diseases, and the complexity of integrating care for complex multimorbidities. Although population ageing is driving the worldwide epidemic of chronic diseases, substantial untapped potential exists to modify the relation between chronological age and health. This objective is especially important for the most age-dependent disorders (ie, dementia, stroke, chronic obstructive pulmonary disease, and vision impairment), for which the burden of disease arises more from disability than from mortality, and for which long-term care costs outweigh health expenditure. The societal cost of these disorders is enormous.


Tobacco Smoke Exposure Reduction Strategies-Do They Work?
Many children experience tobacco smoke exposure (TSE) and parents may take preventive measures to reduce TSE. The study goal is to assess if these strategies are associated with lower cotinine values, an objective biological measure of TSE. Families admitted to Children's Hospital Colorado from 2014 to 2018 who screened positive for TSE were invited to participate in a tobacco smoking cessation/reduction program. Caregivers were consented and asked about demographics, beliefs around smoking, and strategies to reduce TSE. Child urine samples were collected, tested for cotinine levels, and analyzed using geometric means. Bivariable comparisons and multivariable linear regression were completed using SAS v9.4 (SAS Institute, Cary, NC). Two hundred thirteen children (81.4%) are included in this analysis. The median ages of children and parents were 4 and 32 years respectively. Fifty-seven percent of children were male, 36% were Hispanic, and 55% were white. Fifty-six percent of parents had at least some college education and 69% had an annual income less than $50K. The median daily cigarettes smoked per day were 10. Eighty-eight percent reported using at least 1 type of protective measure to prevent TSE and 90% believed they protect other household members from TSE. None of the strategies had a significant relationship with lower cotinine levels on bivariable or multivariable analyses. Parental strategies to decrease TSE did not result in lower cotinine levels. Many measures are not evidence-based and do not protect children. Parent's clothing and homes may create a reservoir for nicotine. Education should focus on exposure elimination and cessation rather than protective measures.


The nature and importance of attachment relationships to parents and peers during adolescence.
The nature and quality of adolescents' attachments to peers and parents were assessed with the newly developed Inventory of Adolescent Attachments. The relative influence on measures of self-esteem and life satisfaction of relations with peers and with parents was then investigated in a hierarchical regression model. The sample consisted of 213 adolescents ranging from 12 to 19 years of age. Two hypotheses were tested: (1) The quality of perceived attachments both to parents and peers would be related to well-being, and (2) the quality of parental relationships would be a more powerful predictor of well-being than would the quality of peer relationships. Confirming the study's hypotheses, the perceived quality of the adolescents' relationships to both peers and parents, their frequency of utilization of peers, and their degree of negative life change were significantly related to both measures of well-being. The quality of attachment to parents was significantly more powerful than that to peers in predicting well-being. In addition, quality of attachment to parents showed a moderating effect under conditions of high life stress on the measures of self-esteem. The study suggests that it is useful to consider the quality of attachments to significant others as an important variable throughout the life span.


Minimally-destructive atmospheric ionisation mass spectrometry authenticates authorship of historical manuscripts.
Authentic historic manuscripts fetch high sums, but establishing their authenticity is challenging, relies on a host of stylistic clues and requires expert knowledge. High resolution mass spectrometry has not, until now, been applied to guide the authentication of historic manuscripts. Robert Burns is a well-known Scottish poet, whose fame, and the eponymous 'Burns Night' are celebrated world-wide. Authenticity of his works is complicated by the 'industrial' production of fakes by Alexander Smith in the 1890s, many of which were of good quality and capable of fooling experts. This study represents the first analysis of the inks and paper used in Burns poetry, in a minimally destructive manner that could find application in many areas. Applying direct infusion mass spectrometry to a panel of selected authenticated Burns and Smith manuscripts, we have produced a Support Vector Machine classifier that distinguishes Burns from Smith with a 0.77 AUC. Using contemporary recipes for inks, we were also able to match features of each to the inks used to produce some of Burns' original manuscripts. We anticipate the method and classifier having broad application in authentication of manuscripts, and our analysis of contemporary inks to provide insights into the production of written works of art.


Photophysics of the galvinoxyl free radical revisited.
The photophysical properties of the free neutral radical galvinoxyl were studied by a combination of femtosecond time-resolved spectroscopy and quantum chemical calculations. The electronic absorption spectrum is dominated by an intense band at 430 nm that is ascribed to the D(9,10)← D(0) transitions. Upon photoexcitation at 400 nm, the population of the D(9,10) states decays within less than 200 fs to the electronic ground state. This ultrafast internal conversion does not involve intramolecular modes with large amplitude motion as the measured dynamics does not show any significant dependence on the environment, but is most probably facilitated by a high density of electronic states of different character. Depending on the solvent, a weak transient band due to the galvinoxylate anion is also observed. This closed-shell species, which is fluorescent although its deactivation is also dominated by non-radiative decay, is generated upon biphotonic ionization of the solvent and electron capture. The ultrashort excited-state lifetime of the galvinoxyl radical precludes photoinduced disproportionation previously claimed to be at the origin of the formation of both anion and cation.


On-Demand Preexposure Prophylaxis in Men at High Risk for HIV-1 Infection.
Antiretroviral preexposure prophylaxis has been shown to reduce the risk of human immunodeficiency virus type 1 (HIV-1) infection in some studies, but conflicting results have been reported among studies, probably due to challenges of adherence to a daily regimen. We conducted a double-blind, randomized trial of antiretroviral therapy for preexposure HIV-1 prophylaxis among men who have unprotected anal sex with men. Participants were randomly assigned to take a combination of tenofovir disoproxil fumarate (TDF) and emtricitabine (FTC) or placebo before and after sexual activity. All participants received risk-reduction counseling and condoms and were regularly tested for HIV-1 and HIV-2 and other sexually transmitted infections. Of the 414 participants who underwent randomization, 400 who did not have HIV infection were enrolled (199 in the TDF-FTC group and 201 in the placebo group). All participants were followed for a median of 9.3 months (interquartile range, 4.9 to 20.6). A total of 16 HIV-1 infections occurred during follow-up, 2 in the TDF-FTC group (incidence, 0.91 per 100 person-years) and 14 in the placebo group (incidence, 6.60 per 100 person-years), a relative reduction in the TDF-FTC group of 86% (95% confidence interval, 40 to 98; P=0.002). Participants took a median of 15 pills of TDF-FTC or placebo per month (P=0.57). The rates of serious adverse events were similar in the two study groups. In the TDF-FTC group, as compared with the placebo group, there were higher rates of gastrointestinal adverse events (14% vs. 5%, P=0.002) and renal adverse events (18% vs. 10%, P=0.03). The use of TDF-FTC before and after sexual activity provided protection against HIV-1 infection in men who have sex with men. The treatment was associated with increased rates of gastrointestinal and renal adverse events. (Funded by the National Agency of Research on AIDS and Viral Hepatitis [ANRS] and others; ClinicalTrials.gov number, NCT01473472.).


Clinical trial experience in Japan and future issues in developing drugs to treat COVID-19.
The National Center for Global Health and Medicine plays a central role in the treatment and research of infectious diseases in Japan. It has conducted various research and development activities on drugs to treat coronavirus disease 2019 (COVID-19) with clinical questions as starting points. Clinical trials are essential in developing new treatment modalities, but we have noticed some characteristic difficulties in clinical trials on emerging and re-emerging infectious diseases. For example, since there is no standard of care when an emerging infectious disease starts to spread, establishing an appropriate control group is complicated, and many things are hurried at the start of trials. This means there is little time to arrange a placebo, and conducting blinded, randomized, controlled trials has been difficult. Another issue characteristic of infectious disease has been that progress in enrolling subjects is affected by the spread of the disease. It was also a struggle to select institutions that provide medical care on the front lines of infectious disease and conduct clinical trials regularly. To start multicenter clinical trials expeditiously, a regulated and structured network is thus considered necessary. From the perspective of implementation, it is preferable to conduct decentralized clinical trials (DCTs) that do not depend on people coming to the medical institution, while from the perspective of preventing infections during the spread of COVID-19, wide adoption of eConsent is desirable. Based on the experience of COVID-19, new measures must be taken to prepare for emerging and re-emerging infectious diseases in the future.


Density gradient multilayer polymerization for creating complex tissue.
An adaptable density gradient multilayer polymerization (DGMP) method facilitates simple fabrication of complex multicompartment scaffolds with structurally continuous interfaces. Solvent density liquid-liquid phase segregation compartmentalizes varied mechanical and chemical cues independently. Bulk photopolymerization produces stratified three-dimensional and two-dimensional matrices. Cells attach to patterned adhesion peptides on biomimetic 2D substrates.


Risk factors for pathologically confirmed benign breast disease.
Between November 1981 and March 1983, data were collected to evaluate risk factors for benign breast lesions in a case-control study based on 288 women with histologically proven benign breast disease, admitted for biopsy to the Tumor Institute of Milan, and 285 age-matched controls. Questions were asked about menstrual and reproductive characteristics, marital status, education, history of various diseases, and lifetime use of oral contraceptives and other hormonal treatments. Nulliparity or low parity, late age at first birth, and late menopause were associated with an increased risk of benign breast disease. The elevated risk associated with late age at first birth was not accounted for by parity. Early age at menarche was associated with an increased risk, but the estimate was not statistically significant. The data do not suggest that the use of oral contraceptives or other female hormones (such as estrogen replacement therapy) is related to the risk of benign breast disease. Risk was apparently lower, however, among current and long-term oral contraceptive users. There was no evidence of a trend with reference to body mass index. The present data indicate a substantial agreement between the risk factors for (pathologically confirmed) benign and malignant breast disease, not only directly, by showing a relationship with parity, age at first birth, and age at menopause, but also indirectly, by failing to produce evidence that greater weight or the use of oral contraceptives has a protective effect.


Labelled drug-related public expenditure in relation to gross domestic product (GDP) in Europe: a luxury good?
"Labelled drug-related public expenditure" is the direct expenditure explicitly labelled as related to illicit drugs by the general government of the state. As part of the reporting exercise corresponding to 2005, the European Monitoring Centre for Drugs and Drug Addiction's network of national focal points set up in the 27 European Union (EU) Member States, Norway, and the candidates countries to the EU, were requested to identify labelled drug-related public expenditure, at the country level. This was reported by 10 countries categorised according to the functions of government, amounting to a total of EUR 2.17 billion. Overall, the highest proportion of this total came within the government functions of Health (66%), and Public Order and Safety (POS) (20%). By country, the average share of GDP was 0.023% for Health, and 0.013% for POS. However, these shares varied considerably across countries, ranging from 0.00033% in Slovakia, up to 0.053% of GDP in Ireland in the case of Health, and from 0.003% in Portugal, to 0.02% in the UK, in the case of POS; almost a 161-fold difference between the highest and the lowest countries for Health, and a 6-fold difference for POS. Why do Ireland and the UK spend so much in Health and POS, or Slovakia and Portugal so little, in GDP terms? To respond to this question and to make a comprehensive assessment of drug-related public expenditure across countries, this study compared Health and POS spending and GDP in the 10 reporting countries. Results found suggest GDP to be a major determinant of the Health and POS drug-related public expenditures of a country. Labelled drug-related public expenditure showed a positive association with the GDP across the countries considered: r = 0.81 in the case of Health, and r = 0.91 for POS. The percentage change in Health and POS expenditures due to a one percent increase in GDP (the income elasticity of demand) was estimated to be 1.78% and 1.23% respectively. Being highly income elastic, Health and POS expenditures can be considered luxury goods; as a nation becomes wealthier it openly spends proportionately more on drug-related health and public order and safety interventions.


Nanog requires BRD4 to maintain murine embryonic stem cell pluripotency and is suppressed by bromodomain inhibitor JQ1 together with Lefty1.
Embryonic stem cells (ESCs) are maintained in an undifferentiated state through expression of the core transcriptional factors Nanog, Oct4, and Sox2. However, the epigenetic regulation of pluripotency is poorly understood. Differentiation of ESCs is accompanied by a global reduction of panacetylation of histones H3 and H4 suggesting that histone acetylation plays an important role in maintenance of ESC pluripotency. Acetylated lysine residues on histones are read by members of the bromodomain family that includes BET (bromodomain and extraterminal domain) proteins for which highly potent and selective inhibitors have been developed. In this study we demonstrate that the pan-BET bromodomain inhibitor JQ1 induces rapid spontaneous differentiation of murine ESCs by inducing marked transcriptional downregulation of Nanog as well as the stemness markers Lefty1 and Lefty2, but not Myc, often used as a marker of BET inhibitor activity in cancer. We show that the effects of JQ1 are recapitulated by knockdown of the BET family member BRD4 implicating this protein in Nanog regulation. These data are also supported by chromatin immunoprecipitation experiments which confirm BRD4 binding at the Nanog promoter that is known to require acetylation by the histone acetyltransferase MOF for transcriptional activity. In further support of our findings, we show that JQ1 antagonizes the stem cell-promoting effects of the histone deacetylase inhibitors sodium butyrate and valproic acid. Our data suggest that BRD4 is critical for the maintenance of ESC pluripotency and that this occurs primarily through the maintenance of Nanog expression.


Identification of two genes with prepilin-like leader sequences involved in type 4 fimbrial biogenesis in Pseudomonas aeruginosa.
Type 4 fimbriae are surface filaments produced by a range of bacterial pathogens for colonization of host epithelial surfaces. In Pseudomonas aeruginosa, they are involved in adhesion as well as in a form of surface translocation called twitching motility, and sensitivity to infection by fimbria-specific bacteriophage. Analysis of the 2.5-kb intergenic region between the previously defined pilR and pilV genes on P. aeruginosa genomic SpeI fragment E has identified three new genes, fimT, fimU, and dadA*. The predicted 18.5-kDa products of the fimT and fimU genes contain prepilin-like leader sequences, whereas the third gene, dadA*, encodes a protein similar to the D-amino acid dehydrogenase of Escherichia coli. Isogenic mutants constructed by allelic exchange demonstrated that the fimU gene was required for fimbrial biogenesis and twitching motility, whereas the fimT and dada* mutants retained wild-type phenotypes. However, overexpression of the fimT gene was found to be able to functionally replace the lack of a fimU gene product, suggesting a subtle role in fimbrial biogenesis. The identification of these proteins increases the similarity between type 4 fimbrial biogenesis and the supersystems involved in macromolecular traffic, such as extracellular protein secretion and DNA uptake, all of which now possess multiple protein species that possess prepilin-like leader sequences.


Microarray-based identification of htrA, a Streptococcus pneumoniae gene that is regulated by the CiaRH two-component system and contributes to nasopharyngeal colonization.
Nasopharyngeal carriage is the reservoir from which most disease with Streptococcus pneumoniae arises. Survival as a commensal in this environment is likely to require a set of adaptations distinct from those needed to cause disease, some of which may be mediated by two-component signal transduction systems (TCSTS). We examined the contributions of nine pneumococcal TCSTS to the process of nasopharyngeal colonization by using an infant rat model. Whereas deletions in all but one of these systems have been associated previously with a high degree of attenuation in a murine model of pneumonia, only the CiaRH system was necessary for efficient carriage. Transcriptional analysis by using microarray hybridization identified a locus consisting of two adjacent genes, htrA and spoJ, that was specifically and strongly downregulated in a DeltaciaRH-null mutant. A S. pneumoniae strain lacking the htrA gene encoding a putative serine protease, but not one lacking spoJ, showed decreased fitness in a competitive model of colonization, a finding consistent with this gene mediating a portion of the carriage deficit observed with the DeltaciaRH strain.


High-energy asymmetric supercapacitors based on free-standing hierarchical Co-Mo-S nanosheets with enhanced cycling stability.
Layered transition metal sulfides (TMS) are emerging as advanced materials for energy storage and conversion applications. In this work, we report a facile and cost-effective anion exchange technique to fabricate a layered, multifaceted, free standing, ultra-thin ternary cobalt molybdenum sulfide nanosheet (Co-Mo-S NS) architecture grown on a 3D porous Ni foam substrate. The unique Co-Mo layered double hydroxides are first synthesized as precursors and consequently transformed into ultra-thin Co-Mo-S NS. When employed as an electrode for supercapacitors, the Co-Mo-S NS delivered an ultra-high specific capacitance of 2343 F g-1 at a current density of 1 mA cm-2 with tremendous rate capability and extraordinary cycling performance (96.6% capacitance retention after 20 000 cycles). Furthermore, assembled Co-Mo-S/nitrogen doped graphene nanosheets (NGNS) in an asymmetric supercapacitor (ASC) device delivered an excellent energy density of 89.6 Wh kg-1, an amazing power density of 20.07 kW kg-1, and superior cycling performance (86.8% capacitance retention after 50 000 cycles). Such exceptional electrochemical performance of Co-Mo-S NS is ascribed to the good electrical contact with the 3D Ni foam, ultra-high contact area with the electrolyte, and enhanced architectural softening during the charging/discharging process. It is expected that the fabricated, unique, ultra-thin Co-Mo-S NS have great potential for future energy storage devices.


Comparative analysis of proteins in the culture supernatants of human intestinal epithelial cells infected with the wild-type and rtxE mutant of Vibrio vulnificus.
Bacterial virulence factors and secreted extracellular proteins from damaged host cells following infection have been recognized as key mediators in the pathophysiological alterations observed in septic shock, and have also been shown to have a synergistic influence on bacterial pathogenicity. We hypothesized that during infections, virulence factors as well as host-shed proteins may synergistically influence aspects of the pathogenicity of V. vulnificus, such as primary septic shock and overproduction of proinflammatory cytokines. However, virulence factors and host-derived proteins have yet to be clearly evaluated during V. vulnificus infection. In this study, we analyzed and compared the proteins in conditioned supernatants generated from co-cultures of host cells and either wild-type or rtxE mutant V. vulnificus using LC-QTOF-MS/MS analysis. In a previous study, we determined that the culture supernatants of the rtxE mutant V. vulnificus-infected INT-407 cells induced significantly lower levels of IL-8 production from human intestinal epithelial cells than did the culture supernatants of wild-type V. vulnificus-infected INT-407 cells. LC-QTOF-MS/MS analysis results demonstrated that levels of proteins such as HSP90 α/β, 14-3-3 γ, PRX II, hnRNP K, β-actin, α-tubulin and V. vulnificus flagellin were significantly lower in the culture supernatants of rtxE mutant V. vulnificus-infected INT-407 cells than in the culture supernatants of wild-type V. vulnificus-infected INT-407 cells. These results demonstrate that V. vulnificus RTX toxins acting via rtxE, a transporter of virulence factors, play a very important role in the pathogenesis of V. vulnificus, as well as in its initial role in inducing pathogenic mediators from host cells.


Stress perfusion imaging using cardiovascular magnetic resonance: a review.
Stress perfusion CMR can provide both excellent diagnostic and important prognostic information in the context of a comprehensive assessment of cardiac anatomy and function. This coupled with the high spatial resolution, and the lack of both attenuation artefacts and ionising radiation, make CMR stress perfusion imaging a highly attractive stress imaging modality. It is now in routine use in many centres, and shows promise in evaluating patients with clinical problems beyond those of epicardial coronary disease.


Acute myocarditis presenting as acute myocardial infarction.
Myocarditis, often caused by an infectious agent, may have a presentation mimicking that of acute myocardial infarction. The authors describe a young man who, when first seen in the emergency department, appeared to have an acute myocardial infarction with normal coronary arteries. At autopsy, the histologic findings were diagnostic of active myocarditis, mixed-cell type. Neutrophilic infiltration with microabscess suggested an infectious cause, although the etiology was uncertain. The patient's clinical course, pathologic findings, and several treatment modalities are discussed. The differentiation between acute myocardial infarction and acute myocarditis can be difficult. Electrocardiographic changes and enzyme elevations are common in both. Therefore, clinicians should consider myocarditis in all patients--particularly young patients--with normal coronary arteries and suspected myocardial infarction.


Cyanobacterial sulfide-quinone reductase: cloning and heterologous expression.
The gene encoding sulfide-quinone reductase (SQR; E.C.1.8.5.'), the enzyme catalyzing the first step of anoxygenic photosynthesis in the filamentous cyanobacterium Oscillatoria limnetica, was cloned by use of amino acid sequences of tryptic peptides as well as sequences conserved in the Rhodobacter capsulatus SQR and in an open reading frame found in the genome of Aquifex aeolicus. SQR activity was also detected in the unicellular cyanobacterium Aphanothece halophytica following sulfide induction, with a V(max) of 180 micromol of plastoquinone-1 (PQ-1) reduced/mg of chlorophyll/h and apparent K(m) values of 20 and 40 microM for sulfide and quinone, respectively. Based on the conserved sequences, the gene encoding A. halophytica SQR was also cloned. The SQR polypeptides deduced from the two cyanobacterial genes consist of 436 amino acids for O. limnetica SQR and 437 amino acids for A. halophytica SQR and show 58% identity and 74% similarity. The calculated molecular mass is about 48 kDa for both proteins; the theoretical isoelectric points are 7.7 and 5.6 and the net charges at a neutral pH are 0 and -14 for O. limnetica SQR and A. halophytica SQR, respectively. A search of databases showed SQR homologs in the genomes of the cyanobacterium Anabaena PCC7120 as well as the chemolithotrophic bacteria Shewanella putrefaciens and Thiobacillus ferrooxidans. All SQR enzymes contain characteristic flavin adenine dinucleotide binding fingerprints. The cyanobacterial proteins were expressed in Escherichia coli under the control of the T7 promoter. Membranes isolated from E. coli cells expressing A. halophytica SQR performed sulfide-dependent PQ-1 reduction that was sensitive to the quinone analog inhibitor 2n-nonyl-4-hydroxyquinoline-N-oxide. The wide distribution of SQR genes emphasizes the important role of SQR in the sulfur cycle in nature.


AIDS: has the problem been adequately assessed?
Epidemiologic inferences about the epidemic of acquired immunodeficiency syndrome (AIDS) are developed from three different sources: case-control studies, cohort studies, and national data on prevalence and incidence. Together these data imply that reducing the number of sexual partners may in itself provide little protection. Shifting to a "lower-risk" behavior will only be helpful if the cumulative risk of transmitting the virus can be reduced nearly to zero. However, if all members of a minimally affected population made such a shift, the benefit to that population could be dramatic. The emerging realization that latency periods are longer than was first thought implies that the epidemic could be much more severe than has been anticipated. Regional trends give no assurance that it has reached a peak, even in areas of highest prevalence. Much more work needs to be done to establish the actual role of saliva in transmitting HTLV-III because oral contagion has the greatest potential to spread the disease. Efforts to determine the latency period are crucial to predicting the future of the epidemic. Preventive measures--behavior modification and education--hold some hope of retarding spread of the epidemic pending development of effective therapy or a vaccine.


Comparing the van Oosterhout and Chybicki-Burczyk methods of estimating null allele frequencies for inbred populations.
In spite of the usefulness of codominant markers in population genetics, the existence of null alleles raises challenging estimation issues in natural populations that are characterized by positive inbreeding coefficients (F > 0). Disregarding the possibility of F > 0 in a population will generally lead to overestimates of null allele frequencies. Conversely, estimates of inbreeding coefficients (F) may be strongly biased upwards (excess homozygotes), in the presence of nontrivial frequencies of null alleles. An algorithm has been presented for the estimation of null allele frequencies in inbred populations (van Oosterhout method), using external estimates of the F-statistics. The goal of this study is to introduce a modification of this method and to provide a formal comparison with an alternative likelihood-based method (Chybicki-Burczyk). Using simulated data, we illustrate the strengths and limitations of these competing methods. Under most circumstances, the likelihood method is preferable, but for highly inbred organisms, a modified van Oosterhout method offers some advantages.


RankProdIt: A web-interactive Rank Products analysis tool.
The first objective of a DNA microarray experiment is typically to generate a list of genes or probes that are found to be differentially expressed or represented (in the case of comparative genomic hybridizations and/or copy number variation) between two conditions or strains. Rank Products analysis comprises a robust algorithm for deriving such lists from microarray experiments that comprise small numbers of replicates, for example, less than the number required for the commonly used t-test. Currently, users wishing to apply Rank Products analysis to their own microarray data sets have been restricted to the use of command line-based software which can limit its usage within the biological community. Here we have developed a web interface to existing Rank Products analysis tools allowing users to quickly process their data in an intuitive and step-wise manner to obtain the respective Rank Product or Rank Sum, probability of false prediction and p-values in a downloadable file. The online interactive Rank Products analysis tool RankProdIt, for analysis of any data set containing measurements for multiple replicated conditions, is available at: http://strep-microarray.sbs.surrey.ac.uk/RankProducts.


Aviremia 10 Years Postdiscontinuation of Antiretroviral Therapy Initiated During Primary Human Immunodeficiency Virus-1 Infection and Association With Gag-Specific T-Cell Responses.
Combination antiretroviral therapy during primary human immunodeficiency virus-1 infection may enable long-term drug-free virological control in rare individuals. We describe a female who maintained aviremia and a normal CD4(+)/CD8(+) T cell ratio for 10 years after stopping therapy, despite a persistent viral reservoir. Cellular immune responses may have contributed to this outcome.


Comparing safety climate between two populations of hospitals in the United States.
To compare safety climate between diverse U.S. hospitals and Veterans Health Administration (VA) hospitals, and to explore the factors influencing climate in each setting. Primary data from surveys of hospital personnel; secondary data from the American Hospital Association's 2004 Annual Survey of Hospitals. Cross-sectional study of 69 U.S. and 30 VA hospitals. For each sample, hierarchical linear models used safety-climate scores as the dependent variable and respondent and facility characteristics as independent variables. Regression-based Oaxaca-Blinder decomposition examined differences in effects of model characteristics on safety climate between the U.S. and VA samples. The range in safety climate among U.S. and VA hospitals overlapped substantially. Characteristics of individuals influenced safety climate consistently across settings. Working in southern and urban facilities corresponded with worse safety climate among VA employees and better safety climate in the U.S. sample. Decomposition results predicted 1.4 percentage points better safety climate in U.S. than in VA hospitals: -0.77 attributable to sample-characteristic differences and 2.2 due to differential effects of sample characteristics. Results suggest that safety climate is linked more to efforts of individual hospitals than to participation in a nationally integrated system or measured characteristics of workers and facilities.


Assessing the impact of COVID-19 border restrictions on dengue transmission in Yunnan Province, China: an observational epidemiological and phylogenetic analysis.
In response to the COVID-19 pandemic, China implemented strict restrictions on cross-border travel to prevent disease importation. Yunnan, a Chinese province that borders dengue-endemic countries in Southeast Asia, experienced unprecedented reduction in dengue, from 6840 recorded cases in 2019 to 260 in 2020. Using a combination of epidemiological and virus genomic data, collected from 2013 to 2020 in Yunnan and neighbouring countries, we conduct a series of analyses to characterise the role of virus importation in driving dengue dynamics in Yunnan and assess the association between recent international travel restrictions and the decline in dengue reported in Yunnan in 2020. We find strong evidence that dengue incidence between 2013-2019 in Yunnan was closely linked with international importation of cases. A 0-2 month lag in incidence not explained by seasonal differences, absence of local transmission in the winter, effective reproductive numbers < 1 (as estimated independently using genetic data) and diverse cosmopolitan dengue virus phylogenies all suggest dengue is non-endemic in Yunnan. Using a multivariate statistical model we show that the substantial decline in dengue incidence observed in Yunnan in 2020 but not in neighbouring countries is closely associated with the timing of international travel restrictions, even after accounting for other environmental drivers of dengue incidence. We conclude that Yunnan is a regional sink for DENV lineage movement and that border restrictions may have substantially reduced dengue burden in 2020, potentially averting thousands of cases. Targeted testing and surveillance of travelers returning from high-risk areas could help to inform public health strategies to minimise or even eliminate dengue outbreaks in non-endemic settings like southern China. Funding for this study was provided by National Key Research and Development Program of China, Beijing Science and Technology Planning Project (Z201100005420010); Beijing Natural Science Foundation (JQ18025); Beijing Advanced Innovation Program for Land Surface Science; National Natural Science Foundation of China (82073616); Young Elite Scientist Sponsorship Program by CAST (YESS) (2018QNRC001); H.T., O.P.G. and M.U.G.K. acknowledge support from the Oxford Martin School. O.J.B was supported by a Wellcome Trust Sir Henry Wellcome Fellowship (206471/Z/17/Z). Chinese translation of the abstract (Appendix 2).


Epidermal growth factor receptor activation in glioblastoma through novel missense mutations in the extracellular domain.
Protein tyrosine kinases are important regulators of cellular homeostasis with tightly controlled catalytic activity. Mutations in kinase-encoding genes can relieve the autoinhibitory constraints on kinase activity, can promote malignant transformation, and appear to be a major determinant of response to kinase inhibitor therapy. Missense mutations in the EGFR kinase domain, for example, have recently been identified in patients who showed clinical responses to EGFR kinase inhibitor therapy. Encouraged by the promising clinical activity of epidermal growth factor receptor (EGFR) kinase inhibitors in treating glioblastoma in humans, we have sequenced the complete EGFR coding sequence in glioma tumor samples and cell lines. We identified novel missense mutations in the extracellular domain of EGFR in 13.6% (18/132) of glioblastomas and 12.5% (1/8) of glioblastoma cell lines. These EGFR mutations were associated with increased EGFR gene dosage and conferred anchorage-independent growth and tumorigenicity to NIH-3T3 cells. Cells transformed by expression of these EGFR mutants were sensitive to small-molecule EGFR kinase inhibitors. Our results suggest extracellular missense mutations as a novel mechanism for oncogenic EGFR activation and may help identify patients who can benefit from EGFR kinase inhibitors for treatment of glioblastoma.


Triage of pregnant women in the emergency department: evaluation of a triage decision aid.
Applying the Australasian Triage Scale to pregnant women presenting to emergency departments (EDs) is difficult as the descriptors may not reflect the urgency of the obstetric condition. This study aimed to examine whether condition-specific algorithms and triage education improved triage assessment and documentation of pregnant women presenting to the ED. Algorithms with a decision aid for triage with minimum agreed descriptors were developed to triage two pregnancy conditions (pre-eclampsia and antepartum haemorrhage). Triage documentation was then audited before (n=50) and after (n=50) a triage education programme which introduced algorithms for both conditions. Significant differences were examined using χ(2) test with significance set at p<0.05. The quality of documentation of specific clinically significant symptoms of pre-eclampsia improved considerably, including the presence of headache from 58% pre-education to 80% post-education (p=0.002), visual disturbances from 58% to 90% (p<0.001), epigastric pain from 24% to 80% (p=0.002) and the presence of fetal movements from 62% to 90% (p=0.001). Documentation of descriptors for vaginal bleeding >20 weeks gestation improved for estimation of blood loss from 54% to 86% (p<0.001), patient 'appearance' from 32% to 62% (p=0.003) and, importantly, descriptions of patient's own assessment of their well-being from 8% to 28% (p=0.009). The introduction of triage education and condition-specific decision aids for triage markedly improved triage assessment and documentation. The application of algorithms may reduce clinical risk resulting from suboptimal triage of pregnant women presenting to EDs.


Short foot exercises have additional effects on knee pain, foot biomechanics, and lower extremity muscle strength in patients with patellofemoral pain.
Patellofemoral pain (PFP) is a common knee problem. The foot posture in a relaxed stance is reported as a distal factor of PFP. However, the effects of short foot exercise (SFE) on the knee and functional factors have not yet been investigated in patients with PFP. This study aimed to investigate the additional effects of SFE on knee pain, foot biomechanics, and lower extremity muscle strength in patients with PFP following a standard exercise program. Thirty patients with a 'weak and pronated' foot subgroup of PFP were randomized into a control group (ConG, n= 15) and a short foot exercise group (SFEG, n= 15) with concealed allocation and blinded to the group assignment. The program of ConG consisted of hip and knee strengthening and stretching exercises. SFEG program consisted of additional SFE. Both groups performed the supervised training protocol two times per week for 6 weeks. Assessment measures were pain visual analog scale (pVAS), Kujala Patellofemoral Score (KPS), navicular drop test (NDT), rearfoot angle (RA), foot posture index (FPI), and strength tests of the lower extremity muscles. Both groups displayed decreases in pVAS scores, but it was only significant in favor of SFEG. NDT, RA, and FPI scores decreased in SFEG whereas they increased in ConG. There was a significant group-by-time interaction effect in hip extensor strength and between-group difference was found to be significantly in favor of SFEG. An intervention program consisting of additional SFE had positive effects on knee pain, navicular position, and rearfoot posture. An increase in the strength of the hip extensors may also be associated with improved stabilization by SFE.


Dynamics of skeletal pattern formation in developing chick limb.
During development of the embryonic chick limb the skeletal pattern is laid out as cartilaginous primordia, which emerge in a proximodistal sequence over a period of 4 days. The differentiation of cartilage is preceded by changes in cellular contacts at specific locations in the precartilage mesenchyme. Under realistic assumptions, the biosynthesis and diffusion through the extracellular matrix of a cell surface protein, such as fibronectin, will lead to spatial patterns of this molecule that could be the basis of the emergent primordia. As cellular differentiation proceeds, the size of the mesenchymal diffusion chamber is reduced in descrete steps, leading to sequential reorganizations of the morphogen pattern. The successive patterns correspond to observed rows of skeletal elements, whose emergence, in theory and in practice, depends on the maintenance of a unique boundary condition at the limb bud apex.


Nonideal ultrathin mantle cloak for electrically large conducting cylinders.
Based on the concept of the scattering cancellation technique, we propose a nonideal ultrathin mantle cloak that can efficiently suppress the total scattering cross sections of an electrically large conducting cylinder (over one free-space wavelength). The cloaking mechanism is investigated in depth based on the Mie scattering theory and is simultaneously interpreted from the perspective of far-field bistatic scattering and near-field distributions. We remark that, unlike the perfect transformation-optics-based cloak, this nonideal cloaking technique is mainly designed to minimize simultaneously several scattering multipoles of a relatively large geometry around considerably broad bandwidth. Numerical simulations and experimental results show that the antiscattering ability of the metasurface gives rise to excellent total scattering reduction of the electrically large cylinder and remarkable electric-field restoration around the cloak. The outstanding cloaking performance together with the good features of and ultralow profile, flexibility, and easy fabrication predict promising applications in the microwave frequencies.


X-ray diffraction evaluation of stress in high pressure deformation experiments.
This paper explores the applicability of x-ray diffraction measurements of stress to high pressure deformation experiments. We model measurements of elastic lattice strains in various geometries for both axial and rotational deformation apparatus. We then show that, for most cases, stresses can be inverted from the diffraction data. A comparison between the results of our models and actual experimental data also indicates that plastic deformation can have an influence that is not addressed properly in the elastic models of lattice strains and should therefore be treated with caution.


A visit to the stroke belt of the United States.
Southeastern part of United States has been called the Stroke Belt due to a much higher incidence of stroke compared to the rest of the country. In this article, I summarize my 2 weeks of observations as a clinical preceptor at the Comprehensive Stroke Center, University of Alabama Hospital, Birmingham, AL. 57 patients were admitted during these 2 weeks, 61% had ischemic strokes, and 23% received intravenous recombinant tissue plasminogen activator (IV rt-PA). Endovascular neuro-interventionalists were performing diagnostic catheter angiography in 14% and emergent revascularization procedures in 7% of consecutive patients. Also, the stroke team enrolled 6 patients into National institute of health (NIH) funded clinical trials (3 Argatroban tPA stroke study (ARTSS), 2 Safety study of external counter pulsation as a treatment for acute ischemic stroke (CUFFS), 1 stenting and aggressive medical management for preventing recurrent stroke in intracranial stenosis (SAMMPRIS). In my opinion, these observations provided me with useful knowledge how to develop a cutting edge, proactive stroke treatment system. In particular, availability 24 × 7 and consistent application of a curative, "finding reasons to treat approach" coupled with state-of the-art technologies and skilled operators could make a huge difference.


Effect of electromagnetic field emitted by cellular phones on fetal heart rate patterns.
The study was planned to determine the effects of electromagnetic fields produced by cellular phones on baseline fetal heart rate, acceleration and deceleration. Forty pregnant women undergoing non-stress test were admitted to the study. Non-stress test was obtained while the subjects were holding the CP on stand by mode and on dialing mode, each for 5 min. Similar recordings were taken while there were no phones around for 10 min. Electromagnetic fields produced by cellular phones do not cause any demonstrable affect in fetal heart rate, acceleration and deceleration.


Impact of community-based interventions for the prevention and control of malaria on intervention coverage and health outcomes for the prevention and control of malaria.
In this paper, we aim to evaluate the effectiveness of community-based interventions (CBIs) for the prevention and management of malaria. We conducted a systematic review and identified 42 studies for inclusion. Twenty-five of the included studies evaluated the impact of the community-based distribution of insecticide-treated nets (ITNs), indoor residual spraying (IRS), or impregnated bed sheets; 14 studies evaluated intermittent preventive therapy (IPT) delivered in community settings; two studies focused on community-based education for malaria prevention; and one study evaluated environmental management through drain cleaning. Our analysis suggests that, overall, the community-based delivery of interventions to prevent and control malaria resulted in a significant increase in ITNs ownership (RR: 2.16, 95% CI: 1.86, 2.52) and usage (RR: 1.77, 95% CI: 1.48, 2.11). However, usage of ITNs was limited to two-thirds of the population who owned them. Community-based strategies also led to a significant decrease in parasitemia (RR: 0.56, 95% CI: 0.42, 0.74), malaria prevalence (RR: 0.46, 95% CI: 0.29, 0.73), malaria incidence (RR: 0.70, 95% CI: 0.54, 0.90), and anemia prevalence (RR: 0.79, 95% CI: 0.64, 0.97). We found a non-significant impact on splenomegaly, birth outcomes (low birth weight, prematurity, stillbirth/miscarriage), anthropometric measures (stunting, wasting, and underweight), and mortality (all-cause and malaria-specific). The subgroup analysis suggested that community-based distribution of ITNs, impregnated bed sheets and IRS, and IPT are effective strategies. Qualitative synthesis suggests that high coverage could be achieved at a lower cost with the integration of CBIs with existing antenatal care and immunization campaigns. Community-based delivery of interventions to prevent and control malaria are effective strategies to improve coverage and access and reduce malaria burden, however, efforts should also be concerted to prevent over diagnosis and drug resistance.


Transplantation after ex vivo lung perfusion: A midterm follow-up.
A large proportion of donor lungs are discarded due to known or presumed organ dysfunction. Ex vivo lung perfusion (EVLP) has proven its value as a tool for discrimination between reversible and irreversible donor lung pathology. However, the long-term outcome after transplantation of lungs after EVLP is essentially unknown. We report short-term and midterm outcomes of recipients who received transplants of EVLP-evaluated lungs. Single-center results of recipients of lungs with prior EVLP were compared with consecutive recipients of non-EVLP lungs (controls) during the same period. Short-term follow-up included time to extubation, time in the intensive care unit, and the presence of primary graft dysfunction at 72 hours postoperatively. Mortality and incidence of chronic lung allograft dysfunction were monitored for up to 4 years after discharge. During a 4-year period, 32 pairs of initially rejected donor lungs underwent EVLP. After EVLP, 22 double lungs and 5 single lungs were subsequently transplanted. During this period, 145 patients received transplants of conventional donor lungs that did not have EVLP and constituted the control group. Median time to extubation was 7 hours in the EVLP group and 6 hours in the non-EVLP control group (p = 0.45). Median intensive care unit stay was 4 days vs. 3 days, respectively (p = 0.15). Primary graft dysfunction grade > 1 was present in 14% in the EVLP group and in 12% in the non-EVLP group at 72 hours after transplant. Survival at 1 year was 92% in the EVLP group and 79% in the non-EVLP group. Cumulative survival and freedom from retransplantation or chronic rejection were also comparable between the 2 groups (p = 0.43) when monitored up to 4 years. Selected donor lungs rejected for transplantation can be used after EVLP. This technique is effective for selection of transplantable donor lungs. Patients who received lungs evaluated under EVLP have short-term and midterm outcomes comparable to recipients of non-EVLP donor lungs.


Factors associated with adolescent overweight/obesity in Ho Chi Minh city.
To identify factors associated with overweight/obesity among adolescents in Ho Chi Minh City, Vietnam. In 2004, a cross-sectional survey was conducted using multi-stage cluster sampling. Weight and height of 2 678 students from 31 junior high schools in Ho Chi Minh City were measured. Dietary intake and physical activity were assessed using a validated food frequency questionnaire and a physical activity questionnaire. Parental, household and neighbourhood characteristics were also collected. Factors associated with overweight/obesity in adolescents were modelled using hierarchical, multivariate logistic regression. "Survey commands" in STATA were used to account for the multi-stage cluster sampling design. After adjusting for other study factors, the odds of overweight/obesity was significantly higher for male gender (OR=3.1, 95% CI: 1.8, 3.8), younger age (OR=3.4, 95% CI: 2.1, 5.3), schools located in wealthy districts (OR=2.5, 95% CI: 1.8, 3.4), higher family economic status (OR=1.9, 95% CI: 1.1, 4.9), higher parental education (OR=2.0, 95% CI: 1.4, 2.9), overweight or obese parents (OR=3.5, 95% CI: 1.7, 7.0), more time spent watching TV (OR=4.3, 95% CI: 1.3, 14.7), frequent consumption of soft drinks (OR=3.0, 95% CI: 1.6, 5.3) and more time studying after class (OR=2.9, 95% CI: 1.7, 4.9). The odds of overweight/obesity was lower with the availability of fruit at home (OR=0.5, 95% CI: 0.3, 0.7), frequent consumption of fruit/vegetables (OR=0.3, 95% CI: 0.2, 0.4) and being physically active (OR=0.5, 95% CI: 0.4, 0.8). Family/parental factors as well as children's lifestyle should be taken into account when planning interventions to prevent adolescent obesity.


Human coproporphyrinogen oxidase. Biochemical characterization of recombinant normal and R231W mutated enzymes expressed in E. coli as soluble, catalytically active homodimers.
To obtain recombinant human coproporphyrinogen oxidase (CPX), a cDNA for the coding region of mature human CPX has been expressed in E. coli. CPX was produced as a fusion protein with glutathione S-transferase followed by the hexapeptide recognition site for thrombin cleavage just preceding first amino acid of the CPX protein. The human CPX was found to be in the soluble fraction. This previously unobtainable human heme synthetic enzyme was purified to electrophoretic homogeneity with a specific activity of 4200 nmol/hr./mg of protein using a Glutathione Sepharose 4B column and gel filtration. Recombinant human CPX exhibits homogeneous behavior during high performance liquid chromatography (HPLC) and the N-terminal sequence, confirmed by protein sequencing, revealed a single polypeptide chain. In its active form, human CPX is a homodimer. According to the hydrodynamic properties derived from analytical ultracentrifugation, dimeric CPX has a nearly globular shape. Additionally, naturally occurring Arg to Trp (R231W)-mutated CPX has been also expressed in E. coli and further characterized. The mutated enzyme has a Km value of 0.55 microM as compared to 0.30 microM for the wild type. The catalytic efficiency (specificity constant, kcat/Km) of the mutated CPX was four fold lower than wild-type enzyme. The activity measurement of the mutated enzyme showed higher thermal sensitivity as compared with wild type CPX. The measured pI for mutated CPX is 5.65, compared to 6.40 for wild type. The pH optima for the mutated and wild-type protein are 6.6 and 6.8, respectively. The R231W mutation of CPX does not affect dimer formation and both normal and mutated CPX exhibit identical sedimentation properties. The thermal denaturation of both wild type and mutant CPX was found to be irreversible. The mutated CPX contained a significant amount of tightly bound porphyrin coproporphyrin. No metal association was found either in wild type or in mutated CPX. The availability of the recombinant human CPX will aid in structural and mechanistic studies.


Is the ovarian reserve influenced by vitamin D deficiency and the dress code in an infertile Iranian population?
In the recent years, vitamin D has become a topical subject and a focus of research not only in reproductive medicine but across many medical disciplines. In reproductive medicine, studies have identified an association between vitamin D status in women and ovarian reserve. In humans, exposure of the skin to sunlight is the main important source of vitamin D. A dress code of wearing concealing clothing is a risk factor for vitamin D deficiency. The objective of this prospective observational study was to evaluate the correlation between vitamin D deficiency and ovarian reserve in a population of infertile women in Iran. As part of the basic fertility assessment of study participants, blood tests were taken to measure vitamin D concentration and transvaginal ultrasound scans were performed on day 2-5 of the cycle to determine antral follicle count (AFC). All study participants were assessed by a reproductive medicine specialist and consultant dermatologist to classify their skin types according to the Fitzpatrick classification. In addition, the dress code of each study participant was recorded noting the percentage of exposed skin not covered by concealing clothing. 189 infertility patients were included in this study. The mean concentration of vitamin D in this study population was 15.46 ng/ml, indicating severe vitamin D deficiency. A statistically significant negative correlation between age and vitamin D (p = 0.008) and age and AFC (p = 0.001) was identified. This study revealed a highly significant correlation between vitamin D concentrations and AFC (p < 0.001). A concealing dress code is an independent risk factor for vitamin D deficiency due to a lack of skin exposure to sunlight. Our study suggests that the so caused severe vitamin D deficiency may play a crucial role in reduced ovarian reserve in the herein described group of an infertile female Iranian population.


Relative Age Effect in Young Swiss Alpine Skiers From 2004 to 2011.
To verify whether relative age effects (RAEs) occur among young male and female Swiss Alpine skiers of different age groups and performance levels. In addition, the efficacy of normalizing performance in physical tests to height and body mass to attenuate RAEs eventually present was tested. The Swiss Ski Power Test consists of anthropometric measures and physical tests for coordination and speed, endurance, and strength and has been used since 2004 to evaluate 11- to 19-y-old Swiss competitive Alpine skiers. The authors analyzed the distribution of 6996 tests performed by 1438 male and 1031 female Alpine skiers between 2004 and 2011 according to the athletes' respective relative age quartiles. Differences in anthropometric measures and performance in physical tests according to quartile were assessed, and the possibility of attenuating eventual RAEs on performance by normalization of results to height and body mass was tested. RAEs were found among all female and male age groups, with no differences between age groups. While performance level did not affect RAE for male skiers, it influenced RAE among female skiers. RAEs also influenced results in all physical tests except upper-limb strength. Normalization of results to body mass attenuated most RAEs identified. Small RAEs are present among young Swiss competitive Alpine skiers and should be taken into account in training and selection settings to prevent the waste of possible future talents. When ranking junior athletes according to their performance in physical tests, normalization of results to body mass decreases the bias caused by RAEs.


A prospective evaluation of the prevalence of submucous cleft palate in patients with isolated cleft lip versus controls.
Although there is an established relationship between cleft lip and overt cleft palate, the relationship between isolated cleft lip and submucous cleft palate has not been investigated. To test the hypothesis that patients with isolated cleft lip have a greater association with submucous cleft palate, a double-armed prospective trial was designed. A study group of 25 consecutive children presenting with an isolated cleft lip, with or without extension through the alveolus but not involving the secondary palate, was compared with a control group of 25 children with no known facial clefts. Eligible patients were examined for the presence of physical criteria associated with classic submucous cleft palate, namely, (1) bifid uvula, (2) absence of the posterior nasal spine, and (3) zona pellucida. Nasoendoscopy was subsequently performed just after induction of general anesthesia, and the findings were correlated with digital palpation of the palatal muscles. Patients who did not satisfy all three physical criteria and in whom nasoendoscopy was distinctly abnormal relative to the control group were classified as having occult submucous cleft palate. Classic submucous cleft palate was found in three study group patients (12 percent), all of whom had flattening or a midline depression of the posterior palate and musculus uvulae on nasoendoscopy and palpable diastasis of the palatal muscles under general anesthesia. An additional six study group patients (24 percent) had similar nasoendoscopic criteria and palpable diastasis of the palatal muscles; they were classified as having occult submucous cleft palate. No submucous cleft palate was identified in the control group. Seventeen patients in the study group had an alveolar cleft with a 53 percent (9 of 17) prevalence of submucous cleft palate. In the present study, classic submucous cleft palate in association with isolated cleft lip was 150 to 600 times the reported prevalence in the general population. All children with an isolated cleft lip should undergo peroral examination and speech/resonance assessment no later than the age of 3 years. Any child with an isolated cleft lip with velopharyngeal inadequacy or before an adenoidectomy should be assessed by flexible nasal endoscopy to avoid missing an occult submucous cleft palate.


Characterization of squamate olfactory receptor genes and their transcripts by the high-throughput sequencing approach.
The olfactory receptor (OR) genes represent the largest multigene family in the genome of terrestrial vertebrates. Here, the high-throughput next-generation sequencing (NGS) approach was applied to characterization of OR gene repertoires in the green anole lizard Anolis carolinensis and the Japanese four-lined ratsnake Elaphe quadrivirgata. Tagged polymerase chain reaction (PCR) products amplified from either genomic DNA or cDNA of the two species were used for parallel pyrosequencing, assembling, and screening for errors in PCR and pyrosequencing. Starting from the lizard genomic DNA, we accurately identified 56 of 136 OR genes that were identified from its draft genome sequence. These recovered genes were broadly distributed in the phylogenetic tree of vertebrate OR genes without severe biases toward particular OR families. Ninety-six OR genes were identified from the ratsnake genomic DNA, implying that the snake has more OR gene loci than the anole lizard in response to an increased need for the acuity of olfaction. This view is supported by the estimated number of OR genes in the Burmese python's draft genome (∼280), although squamates may generally have fewer OR genes than terrestrial mammals and amphibians. The OR gene repertoire of the python seems unique in that many class I OR genes are retained. The NGS approach also allowed us to identify candidates of highly expressed and silent OR gene copies in the lizard's olfactory epithelium. The approach will facilitate efficient and parallel characterization of considerable unbiased proportions of multigene family members and their transcripts from nonmodel organisms.


Prevalence, diagnosis, and disease course of pertussis in adults with acute cough: a prospective, observational study in primary care.
Most cases of adult pertussis probably remain undiagnosed. To explore the prevalence, diagnosis, and disease course of acute pertussis infection in adult patients presenting with acute cough. Prospective observational study between 2007 and 2010 in primary care in 12 European countries. Adults presenting with acute cough (duration of ≤28 days) were included. Bordetella pertussis infection was determined by polymerase chain reaction (from nasopharyngeal flocked swabs and sputa) and by measurement of immunoglobulin G antibodies to pertussis toxin (PT) in venous blood at day 28. An antibody titre to PT of ≥125 IU/ml or PCR positive result in a respiratory sample defined recent infection. Patients completed a symptom diary for 28 days. Serum and/or respiratory samples were obtained in 3074 patients. Three per cent (93/3074) had recent B. pertussis infection. Prior cough duration >2 weeks discriminated to some extent between those with and without pertussis (adjusted odds ratio 1.89, 95% confidence interval = 1.17 to 3.07; P = 0.010). Median cough duration after presentation was 17 and 12 days in patients with and without pertussis, respectively (P = 0.008). Patients with pertussis had longer duration of phlegm production (P = 0.010), shortness of breath (P = 0.037), disturbed sleep (P = 0.013) and interference with normal activities or work (P = 0.033) after presentation. Pertussis infection plays a limited role among adults presenting with acute cough in primary care, but GPs should acknowledge the possibility of pertussis in uncomplicated lower respiratory tract infection. As in children, pertussis also causes prolonged symptoms in adults. However, pertussis is difficult to discern from other acute cough syndromes in adults at first presentation.


Fixation biases affecting human SNPs.
Under neutrality all classes of mutation have an equal probability of becoming fixed in a population. In this article, we describe our analysis of the frequency distributions of >5000 human SNPs and provide evident of biases in the process of fixation of certain classes of point mutation that are most likely to be attributable to biased gene conversion. The results indicate an increased fixation probability of mutations that result in the incorporation of a GC base pair. Furthermore, in transcribed regions this process exhibits strand asymmetry, and is biased towards preserving a G base on the coding strand. Biased gene conversion has the potential to explain both existence of isochores and the compositional asymmetry in mammalian transcribed regions.


Bias analysis applied to Agricultural Health Study publications to estimate non-random sources of uncertainty.
The associations of pesticide exposure with disease outcomes are estimated without the benefit of a randomized design. For this reason and others, these studies are susceptible to systematic errors. I analyzed studies of the associations between alachlor and glyphosate exposure and cancer incidence, both derived from the Agricultural Health Study cohort, to quantify the bias and uncertainty potentially attributable to systematic error. For each study, I identified the prominent result and important sources of systematic error that might affect it. I assigned probability distributions to the bias parameters that allow quantification of the bias, drew a value at random from each assigned distribution, and calculated the estimate of effect adjusted for the biases. By repeating the draw and adjustment process over multiple iterations, I generated a frequency distribution of adjusted results, from which I obtained a point estimate and simulation interval. These methods were applied without access to the primary record-level dataset. The conventional estimates of effect associating alachlor and glyphosate exposure with cancer incidence were likely biased away from the null and understated the uncertainty by quantifying only random error. For example, the conventional p-value for a test of trend in the alachlor study equaled 0.02, whereas fewer than 20% of the bias analysis iterations yielded a p-value of 0.02 or lower. Similarly, the conventional fully-adjusted result associating glyphosate exposure with multiple myleoma equaled 2.6 with 95% confidence interval of 0.7 to 9.4. The frequency distribution generated by the bias analysis yielded a median hazard ratio equal to 1.5 with 95% simulation interval of 0.4 to 8.9, which was 66% wider than the conventional interval. Bias analysis provides a more complete picture of true uncertainty than conventional frequentist statistical analysis accompanied by a qualitative description of study limitations. The latter approach is likely to lead to overconfidence regarding the potential for causal associations, whereas the former safeguards against such overinterpretations. Furthermore, such analyses, once programmed, allow rapid implementation of alternative assignments of probability distributions to the bias parameters, so elevate the plane of discussion regarding study bias from characterizing studies as "valid" or "invalid" to a critical and quantitative discussion of sources of uncertainty.


Cholesterol Modification of an Anticancer Drug for Efficient Incorporation into a Supramolecular Hydrogel System.
Treatment of cancer in the peritoneal cavity may be improved with macroscale drug delivery systems that offer control over intraperitoneal concentration of chemotherapeutic agents. Currently, suitable drug carriers to facilitate a sustained release of small hydrophilic drugs such as mitomycin C are lacking. For this purpose, a pH-responsive supramolecular hydrogel based on ureido-pyrimidinone (UPy) chemistry is utilized here. In order to provide a sustained release profile, a lipophilicity-increasing cholesterol conjugation strategy is proposed that enhances affinity between the modified drug (mitomycin-PEG24 -cholesterol, MPC) and the hydrophobic compartments in the UPy gel. Additional advantages of cholesterol conjugation include improved chemical stability and potency of mitomycin C. In vitro the tunability of the system to obtain optimal effective concentrations over time is demonstrated with a combinatorial treatment of mitomycin C and MPC in one UPy hydrogel delivery system.