The assessment of data sources for influenza virologic surveillance in New York State.
Following the 2013 USA release of the Influenza Virologic Surveillance Right Size Roadmap, the New York State Department of Health (NYSDOH) embarked on an evaluation of data sources for influenza virologic surveillance. To assess NYS data sources, additional to data generated by the state public health laboratory (PHL), which could enhance influenza surveillance at the state and national level. Potential sources of laboratory test data for influenza were analyzed for quantity and quality. Computer models, designed to assess sample sizes and the confidence of data for statistical representation of influenza activity, were used to compare PHL test data to results from clinical and commercial laboratories, reported between June 8, 2013 and May 31, 2014. Sample sizes tested for influenza at the state PHL were sufficient for situational awareness surveillance with optimal confidence levels, only during peak weeks of the influenza season. Influenza data pooled from NYS PHLs and clinical laboratories generated optimal confidence levels for situational awareness throughout the influenza season. For novel influenza virus detection in NYS, combined real-time (rt) RT-PCR data from state and regional PHLs achieved ≥85% confidence during peak influenza activity, and ≥95% confidence for most of low season and all of off-season. In NYS, combined data from clinical, commercial, and public health laboratories generated optimal influenza surveillance for situational awareness throughout the season. Statistical confidence for novel virus detection, which is reliant on only PHL data, was achieved for most of the year.


Does "supported decision-making" in India's Mental Health Care Bill, 2013, measure up to the CRPD's standards?
The Convention on the Rights of Persons with Disabilities (CRPD) requires States to replace their mental health laws based on substitute decision-making for persons with mental health issues with laws based on the supported decision-making paradigm. However, the exact scope of the CRPD's mandates is currently under debate, especially in the case of persons with very high support needs. The Mental Health Care Bill, 2013, introduces supported decision-making in India in the form of advance directives and nominated representatives. This article discusses how far the Bill measures up to the CRPD's standards and highlights some of the difficulties when the support needs of the person are very high.


Melatonin increases brown adipose tissue mass and function in Zücker diabetic fatty rats: implications for obesity control.
Melatonin limits obesity in rodents without affecting food intake and activity, suggesting a thermogenic effect. Previously we demonstrated that melatonin browns subcutaneous fat in Zücker diabetic fatty (ZDF) rats. Other works pointed to melatonin as a signal that increases brown adipose tissue (BAT) mass and function in rodents. However, direct proof of thermogenic properties (uncoupled mitochondria) of the newly recruited BAT in response to melatonin is still lacking. Therefore, in this work, we investigated if melatonin recruits thermogenic BAT in ZDF rats. Zücker lean (ZL) and ZDF animals were subdivided into two groups, control (C) and treated with oral melatonin (M) for 6 weeks. Mitochondrial mass, activity of citrate synthase (CS), and respiratory chain complexes I and IV were lower in C-ZDF than in C-ZL animals (P < .001). Melatonin treatment increased BAT weight in ZDF rats (P < .001). Also, it rose mitochondrial mass (P < .01) and activities of CS and complexes I and IV (P < .001) in both, ZDF and ZL rats. Uncoupling protein 1 (UCP1) mRNA and protein were 50% lower in BAT from obese rats. Also, guanosine diphosphate (GDP) binding was lower in ZDF than in lean rats (P < .01). Melatonin treatment of obese rats restored the expression of UCP1 and GDP binding to levels of lean rats and sensitized the thermogenic response to cold exposure. These data demonstrated that melatonin recruits thermogenic BAT in ZDF rats. This may contribute to melatonin's control of body weight and its metabolic benefits.


The dialectics of childhood diarrhea mortality.
As in European countries a century ago, diarrhea is a major cause of child mortality in poor countries today. In Stockholm at the turn of the 19th century, political commitment, infrastructural investments in water and sanitation, and enforcement of sanitary improvements by a strong implementing organization helped eliminate diarrhea as a principal cause of death among children. These interventions also had an equitable impact on social class differences in diarrhea mortality, but not on overall mortality; overall mortality declined, but class differences remained. General infrastructural improvement and health education coupled with targeted interventions to vulnerable children may be successful in improving child health and reducing social differentials in mortality. Specific health care interventions may need to be complemented by infrastructural investments to improve water and sanitation if diarrhea mortality is to be further reduced in poor countries today.


Patterns of GH output and their synchrony with short-term height increments influence stature and growth performance in normal children.
GH is the principal hormone driving growth throughout childhood, but the mechanism by which changing GH secretion is translated into height (Ht) remains undefined. We previously demonstrated that variability in urinary GH (uGH) output over weeks is an important statistical determinant of stature in normal children. We now examine the relationship between the temporal pattern of uGH output and growth using approximate entropy (ApEn) in 29 healthy prepubertal children, monitored serially three times a week from September to June [13 males and 16 females; age, 5.7-7.8 yr; Ht SD score, -2.9 to +2.3; median, 85 measurements per subject]. ApEn is a measure of process irregularity, with low values indicating strong regularity or subpattern persistence and high values indicating greater irregularity. The bivariate extension of this method, cross (X)-ApEn, provides a measure of the strength of interaction between two paired time series (joint signal synchrony). Low values of X-ApEn indicate synchrony, whereas high values indicate asynchrony. Weekly Ht gain, weight (Wt) gain, and uGH output showed significant orderliness in 20 of 29, 26 of 29, and 29 of 29 subjects, respectively, with weekly changes in uGH output being significantly more ordered than Ht gain (P < 0.001) and Wt gain (P < 0.01). ApEn(GH) was positively correlated with Ht gain over the whole year (r = 0.42) and Ht velocity SD score (r = 0.42; both, P < 0.05). Thus, increased irregularity in successive weekly GH output was associated with increased Ht gain and growth rate. There was significant synchrony between Ht and uGH, Wt and uGH, and Ht and Wt in 16 of 29, 22 of 29, and 25 of 29 subjects, respectively, as measured by X-ApEn. The degree of synchrony between Ht and GH output [X-ApEn(Ht:GH)] was inversely correlated with Ht SD score (r = -0.45) and Ht velocity SD score (r = -0.40; both P < 0.05), indicating that increased joint signal synchrony between Ht and GH accompanies tall stature and increased growth rate. These results extend the concept that variability in GH output over time is an important determinant of growth in normal children by indicating that an irregular temporal pattern of GH contributes to growth. Furthermore, synchrony between GH output and short-term Ht increments is associated with tall stature and good growth.


Selective hematopoietic stem cell ablation using CD117-antibody-drug-conjugates enables safe and effective transplantation with immunity preservation.
Hematopoietic stem cell transplantation (HSCT) is a curative therapy for blood and immune diseases with potential for many settings beyond current standard-of-care. Broad HSCT application is currently precluded largely due to morbidity and mortality associated with genotoxic irradiation or chemotherapy conditioning. Here we show that a single dose of a CD117-antibody-drug-conjugate (CD117-ADC) to saporin leads to > 99% depletion of host HSCs, enabling rapid and efficient donor hematopoietic cell engraftment. Importantly, CD117-ADC selectively targets hematopoietic stem cells yet does not cause clinically significant side-effects. Blood counts and immune cell function are preserved following CD117-ADC treatment, with effective responses by recipients to both viral and fungal challenges. These results suggest that CD117-ADC-mediated HSCT pre-treatment could serve as a non-myeloablative conditioning strategy for the treatment of a wide range of non-malignant and malignant diseases, and might be especially suited to gene therapy and gene editing settings in which preservation of immunity is desired.


Chiral auxiliary recycling in continuous flow: automated recovery and reuse of Oppolzer's sultam.
The telescoping of a three-stage, chiral auxiliary-mediated transformation in flow is described, including continuous separation of the product and auxiliary. The auxiliary can either be collected for later reuse, or directly fed back to the beginning of the process for recycling in real time, enabling each molecule of auxiliary to make multiple equivalents of chiral product and thus minimizing the step- and atom-economy issues associated with auxiliary-mediated synthesis. This concept is demonstrated for the asymmetric hydrogenation of olefins using Oppolzer's sultam, shortening the total reaction time >100 fold compared to batch, and demonstrating formal sub-stoichiometric auxiliary loading with respect to the process by automating auxiliary recycling within a closed loop.


Gene expression profiling reveals the defining features of the classical, intermediate, and nonclassical human monocyte subsets.
New official nomenclature subdivides human monocytes into 3 subsets: the classical (CD14(++)CD16(-)), intermediate (CD14(++)CD16(+)), and nonclassical (CD14(+)CD16(++)) monocytes. This introduces new challenges, as monocyte heterogeneity is mostly understood based on 2 subsets, the CD16(-) and CD16(+) monocytes. Here, we comprehensively defined the 3 circulating human monocyte subsets using microarray, flow cytometry, and cytokine production analysis. We find that intermediate monocytes expressed a large majority (87%) of genes and surface proteins at levels between classical and nonclassical monocytes. This establishes their intermediary nature at the molecular level. We unveil the close relationship between the intermediate and nonclassic monocytes, along with features that separate them. Intermediate monocytes expressed highest levels of major histocompatibility complex class II, GFRα2 and CLEC10A, whereas nonclassic monocytes were distinguished by cytoskeleton rearrangement genes, inflammatory cytokine production, and CD294 and Siglec10 surface expression. In addition, we identify new features for classic monocytes, including AP-1 transcription factor genes, CLEC4D and IL-13Rα1 surface expression. We also find circumstantial evidence supporting the developmental relationship between the 3 subsets, including gradual changes in maturation genes and surface markers. By comprehensively defining the 3 monocyte subsets during healthy conditions, we facilitate target identification and detailed analyses of aberrations that may occur to monocyte subsets during diseases.


Physical activity and preventable premature deaths from non-communicable diseases in Brazil.
Studies on the impact of counterfactual scenarios of physical activity on premature deaths from non-communicable diseases (NCDs) are sparse in the literature. We estimated preventable premature deaths from NCDs (diabetes, ischemic heart disease, stroke, and breast and colon cancers) in Brazil by increasing population-wide physical activity (i) to theoretical minimum risk exposure levels; (ii) reaching the physical activity recommendation; (iii) reducing insufficient physical activity by 10%; and (iv) eliminating the gender differences in physical activity. Preventable fractions were estimated using data from a nationally representative survey, relative risks from a meta-analysis and number of premature deaths (30-69 years) from the Brazilian Mortality Information System. Physical activity could potentially avoid up to 16 700 premature deaths from NCDs in Brazil, corresponding to 5.75 and 3.23% of premature deaths from major NCDs and of all-causes, respectively. Other scenarios suggested the following impact on premature deaths: reaching physical activity recommendation (5000 or 1.74% of major NCDs); 10% reduction in insufficient physical activity (500 or 0.17% of major NCDs); eliminating gender differences in physical activity (1000 or 0.33% of major NCDs). Physical activity may play an important role to reduce premature deaths from NCD in Brazil.


Rhodococcus olei sp. nov., with the ability to degrade petroleum oil, isolated from oil-contaminated soil.
A Gram-stain-positive, non-motile, creamy-white and rod-coccus shaped actinobacterium, designated strain Ktm-20T, capable of degrading petroleum oil was isolated from oil-contaminated soil. Strain Ktm-20T was able to grow at 15-37 °C, at pH 5.5-10.0 and at 0.0-2.0 % (w/v) NaCl concentration. This strain was taxonomically characterized by a polyphasic approach. The 16S rRNA gene sequence analysis showed that strain Ktm-20T belonged to the genus Rhodococcus and is closely related to Rhodococcus triatomae DSM 44892T, Rhodococcus pedocola UC12T, Rhodococcus wratislaviensis NBRC 100605T, Rhodococcus agglutinans CFH S0262T and Rhodococcus canchipurensis MBRL 353T (98.8, 98.7, 98.5, 98.4 and 98.3 % gene sequence similarity, respectively). The only respiratory quinone was MK-8(H2); the major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside; and the predominant fatty acids were C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and C18 : 1ω9c. The cell-wall peptidoglycan contained meso-diaminopimelic acid; and galactose, glucose, arabinose and ribose were detected as diagnostic sugars from whole-cell hydrolysates. Mycolic acids were detected. The DNA G+C content was 70.9 mol%. The DNA-DNA relatedness values between strain Ktm-20T and closely related species of the genus Rhodococcus were between 38.3-25.3 %, which falls below the threshold value of 70 % for the strain to be considered as novel. The morphological, physiological, chemotaxonomic and phylogenetic analyses clearly distinguished this strain from its closest phylogenetic neighbours. Thus, strain Ktm-20T represents a novel species of the genus Rhodococcus, for which the name Rhodococcus olei sp. nov. is proposed. The type strain is Ktm-20T (=KEMB 9005-695T=KACC 19390T=JCM 32206T).


Increased levels of the oxidative stress biomarker 8-iso-prostaglandin F2α in wastewater associated with tobacco use.
Wastewater analysis has been demonstrated to be a complementary approach for assessing the overall patterns of drug use by a population while the full potential of wastewater-based epidemiology has yet to be explored. F2-isoprostanes are a prototype wastewater biomarker to study the cumulative oxidative stress at a community level. In this work, 8-iso-prostaglandin F2α (8-iso-PGF2α) was analysed in raw 24 h-composite wastewater samples collected from 4 Norwegian and 7 other European cities in 2014 and 2015. Using the same samples, biomarkers of alcohol (ethyl sulfate) and tobacco (trans-3'-hydroxycotinine) use were also analysed to investigate any possible correlation between 8-iso-PGF2α and the consumption of the two drugs. The estimated per capita daily loads of 8-iso-PGF2α in the 11 cities ranged between 2.5 and 9.9 mg/day/1000 inhabitants with a population-weighted mean of 4.8 mg/day/1000 inhabitants. There were no temporal trends observed in the levels of 8-iso-PGF2α, however, spatial differences were found at the inter-city level correlating to the degree of urbanisation. The 8-iso-PGF2α mass load was found to be strongly associated with that of trans-3'-hydroxycotinine while it showed no correlation with ethyl sulfate. The present study shows the potential for 8-iso-PGF2α as a wastewater biomarker for the assessment of community public health.


Genetic background, and not ontogenetic effects, affects avian seasonal timing of reproduction.
Avian seasonal timing is a life-history trait with important fitness consequences and which is currently under directional selection due to climate change. To predict micro-evolution in this trait, it is crucial to properly estimate its heritability. Heritabilities are often estimated from pedigreed wild populations. As these are observational data, it leaves the possibility that the resemblance between related individuals is not due to shared genes but to ontogenetic effects; when the environment for the offspring provided by early laying pairs differs from that by late pairs and the laying dates of these offspring when they reproduce themselves is affected by this environment, this may lead to inflated heritability estimates. Using simulation studies, we first tested whether and how much such an early environmental effect can inflate heritability estimates from animal models, and we showed that pedigree structure determines by how much early environmental effects inflate heritability estimates. We then used data from a wild population of great tits (Parus major) to compare laying dates of females born early in the season in first broods and from sisters born much later, in second broods. These birds are raised under very different environmental conditions but have the same genetic background. The laying dates of first and second brood offspring do not differ when they reproduce themselves, clearly showing that ontogenetic effects are very small and hence, family resemblance in timing is due to genes. This finding is essential for the interpretation of the heritabilities reported from wild populations and for predicting micro-evolution in response to climate change.


Improving Detection Accuracy of Memristor-Based Bio-Signal Sensing Platform.
Recently a novel neuronal activity sensor exploiting the intrinsic thresholded integrator capabilities of memristor devices has been proposed. Extracellular potentials captured by a standard bio-signal acquisition platform are fed into a memristive device which reacts to the input by changing its resistive state (RS) only when the signal ampitude exceeds a threshold. Thus, significant peaks in the neural signal can be stored as non-volatile changes in memristor resistive state whilst noise is effectively suppressed. However, as a memristor is subjected to increasing numbers of supra-threshold stimuli during practical operation, it accumulates (RS) changes and eventually saturates. This leads to severely redsignaluced neural activity detection capabilities. In this work we explore different signal processing and memristor operating procedure strategies in order to improve the detection rate of significant neuronal activity events. We analyse the data obtained from a single-memristive device biased with a reference neural recording and observe that performance can be improved markedly by a) increasing the frequency at which the memristor is reset to an initial resistive state where it is known to be highly responsive, b) appropriately preconditioning the input waveform through application of gain and offset in order to optimally exploit the intrinsic device behaviour. All results are validated by benchmarking obtained spike detection performance against a state-of-the-art template matching system utilising computationally-heavy, multi-dimensional, principal component analysis.


WHO analysis of causes of maternal death: a systematic review.
The reduction of maternal deaths is a key international development goal. Evidence-based health policies and programmes aiming to reduce maternal deaths need reliable and valid information. We undertook a systematic review to determine the distribution of causes of maternal deaths. We selected datasets using prespecified criteria, and recorded dataset characteristics, methodological features, and causes of maternal deaths. All analyses were restricted to datasets representative of populations. We analysed joint causes of maternal deaths from datasets reporting at least four major causes (haemorrhage, hypertensive disorders, sepsis, abortion, obstructed labour, ectopic pregnancy, embolism). We examined datasets reporting individual causes of death to investigate the heterogeneity due to methodological features and geographical region and the contribution of haemorrhage, hypertensive disorders, abortion, and sepsis as causes of maternal death at the country level. 34 datasets (35,197 maternal deaths) were included in the primary analysis. We recorded wide regional variation in the causes of maternal deaths. Haemorrhage was the leading cause of death in Africa (point estimate 33.9%, range 13.3-43.6; eight datasets, 4508 deaths) and in Asia (30.8%, 5.9-48.5; 11,16 089). In Latin America and the Caribbean, hypertensive disorders were responsible for the most deaths (25.7%, 7.9-52.4; ten, 11,777). Abortion deaths were the highest in Latin America and the Caribbean (12%), which can be as high as 30% of all deaths in some countries in this region. Deaths due to sepsis were higher in Africa (odds ratio 2.71), Asia (1.91), and Latin America and the Caribbean (2.06) than in developed countries. Haemorrhage and hypertensive disorders are major contributors to maternal deaths in developing countries. These data should inform evidence-based reproductive health-care policies and programmes at regional and national levels. Capacity-strengthening efforts to improve the quality of burden-of-disease studies will further validate future estimates.


Interactive effects of early and later nutritional conditions on the adult antioxidant defence system in zebra finches.
In vertebrates, antioxidant defences comprise a mixture of endogenously produced components and exogenously obtained antioxidants that are derived mostly from the diet. It has been suggested that early-life micronutritional conditions might influence the way in which the antioxidant defence system operates, which could enable individuals to adjust the activity of the endogenous and exogenous components in line with their expected intake of dietary antioxidants if the future environment resembles the past. We investigated this possibility by experimentally manipulating the micronutrient content of the diet during different periods of postnatal development in the zebra finch (Taeniopygia guttata). Birds that had a low micronutrient diet during the growth phase initially had a lower total antioxidant capacity (TAC) than those reared under a high micronutrient diet, but then showed a compensatory response, so that by the end of the growth phase, the TAC of the two groups was the same. Interestingly, we found an interactive effect of micronutrient intake early and late in development: only those birds that continued with the same dietary treatment (low or high) throughout development showed a significant increase in their TAC during the period of sexual maturation. A similar effect was also found in the level of enzymatic antioxidant defences (glutathione peroxidase; GPx). No significant effects were found in the level of oxidative damage in lipids [malondialdehyde (MDA) levels]. These findings demonstrate the importance of early and late developmental conditions in shaping multiple aspects of the antioxidant system. Furthermore, they suggest that young birds may adjust their antioxidant defences to enable them to 'thrive' on diets rich or poor in micronutrients later in life.


Explanation of the discrepancy between the degree of organic nitrate decomposition, nitrite formation and guanylate cyclase stimulation.
We continuously studied the quantitative formation of nitric oxide (NO), nitrite and nitrate ions from several organic nitrate esters in the presence of various thiol-containing compounds by spectroscopy and HPLC. The results indicate that there are different pathways of decomposition depending on the chemical nature of the mercaptan tested. The amino acid cysteine is known to function as an essential cofactor for guanylate cyclase activation by organic nitrates in vitro. For comparison we investigated several structural analogues with respect to their nitric oxide or nitrite ion releasing potency. Both were found to represent the main products resulting from nitrate ester breakdown besides the respective alcohols. We found that only those compounds were able to activate the enzyme in the presence of nitroglycerin (GTN) which induce the release of NO as well. On the other hand, nearly all other thiols tested caused an in vitro decomposition of organic nitrates by producing excess nitrite and the corresponding disulfide without the formation of NO. Thus, the decomposition of organic nitrates to nitrite ions does not contribute at all to activation of guanylate cyclase. Our results confirm that the liberation of nitric oxide is the common principle of action for all nitrovasodilators. In addition, our results suggest that the thiol consuming transformation of organic nitrates into nitrite ions (ratio NO/nitrite 1:10) may lead to a depletion of cysteine stores, resulting in a decreased formation of NO and, consequently, in a decrease of guanylate cyclase activation, clinically arising as nitrate tolerance.


Measuring quality of life of people with predementia and dementia and their caregivers: a systematic review protocol.
Dementia is the fastest growing major cause of disability globally and may have a profound impact on the health-related quality of life (HRQoL) of both the patient with dementia and those who care for them. This review aims to systematically identify and synthesise the measurements of HRQoL for people with, and their caregivers across the full spectrum of, dementia from its preceding stage of predementia to end of life. A systematic literature review was conducted in Medical Literature Analysis and Retrieval System Online , ExcerptaMedicadataBASE, Cochrane Database of Systematic Reviews , Cochrane Central Register of Controlled Trials, Database of Abstracts of Reviews of Effect, National Health Service Economic Evaluation Database and PsycINFO between January 1990 and the end of April 2017. Two reviewers will independently assess each study for inclusion and disagreements will be resolved by a third reviewer. Data will be extracted using a predefined data extraction form following best practice. Study quality will be assessed with the Effective Public Health Practice Project quality assessment tool. HRQoL measurements will be presented separately for people with dementia and caregivers by instrument used and, when possible, HRQoL will be reported by disease type and stage of the disease. Descriptive statistics of the results will be provided. A narrative synthesis of studies will also be provided discussing differences in HRQoL measurements by instrument used to estimate it, type of dementia and disease severity. This systematic literature review is exempt from ethics approval because the work is carried out on published documents. The findings of the review will be disseminated in a related peer-reviewed journal and presented at conferences. They will also contribute to the work developed in the Real World Outcomes across the Alzheimer's disease spectrum for better care: multimodal data access platform (ROADMAP). CRD42017071416.


Formation and release of arrestin domain-containing protein 1-mediated microvesicles (ARMMs) at plasma membrane by recruitment of TSG101 protein.
Mammalian cells are capable of delivering multiple types of membrane capsules extracellularly. The limiting membrane of late endosomes can fuse with the plasma membrane, leading to the extracellular release of multivesicular bodies (MVBs), initially contained within the endosomes, as exosomes. Budding viruses exploit the TSG101 protein and endosomal sorting complex required for transport (ESCRT) machinery used for MVB formation to mediate the egress of viral particles from host cells. Here we report the discovery of a virus-independent cellular process that generates microvesicles that are distinct from exosomes and which, like budding viruses, are produced by direct plasma membrane budding. Such budding is driven by a specific interaction of TSG101 with a tetrapeptide PSAP motif of an accessory protein, arrestin domain-containing protein 1 (ARRDC1), which we show is localized to the plasma membrane through its arrestin domain. This interaction results in relocation of TSG101 from endosomes to the plasma membrane and mediates the release of microvesicles that contain TSG101, ARRDC1, and other cellular proteins. Unlike exosomes, which are derived from MVBs, ARRDC1-mediated microvesicles (ARMMs) lack known late endosomal markers. ARMMs formation requires VPS4 ATPase and is enhanced by the E3 ligase WWP2, which interacts with and ubiquitinates ARRDC1. ARRDC1 protein discharged into ARMMs was observed in co-cultured cells, suggesting a role for ARMMs in intercellular communication. Our findings reveal an intrinsic cellular mechanism that results in direct budding of microvesicles from the plasma membrane, providing a formal paradigm for the evolutionary recruitment of ESCRT proteins in the release of budding viruses.


Polymorphisms in calpastatin and mu-calpain genes are associated with beef iron content.
The objective of this study was to assess the association of markers in the calpastatin and mu-calpain loci with iron in beef cattle muscle. The population consisted of 259 cross-bred steers from Beefmaster, Brangus, Bonsmara, Romosinuano, Hereford and Angus sires. Total iron and heme iron concentrations were measured. Markers in the calpastatin (referred to as CAST) and mu-calpain (referred to as CAPN4751) genes were used to assess their association with iron levels. The mean and standard error for iron and heme iron content in the population was 35.6 ± 1.3 μg and 27.1 ± 1.4 μg respectively. Significant associations (P < 0.01) of markers were observed for both iron and heme iron content. For CAST, animals with the CC genotype had higher levels of iron and heme iron in longissimus dorsi muscle. For CAPN4751, individuals with the TT genotype had higher concentrations of iron and heme iron than did animals with the CC and CT genotypes. Genotypes known to be associated with tougher meat were associated with higher levels of iron concentration.


The puff-specific Drosophila protein Bj6, encoded by the gene no-on transient A, shows homology to RNA-binding proteins.
The DNA coding for the puff-specific protein Bj6 has been isolated by expression cloning. The gene is localized in 14C1,2 on the X chromosome and is expressed ubiquitously during embryonic development with prominent expression during the first 12 h of embryogenesis. cDNA and genomic clones have been sequenced and show a single open-reading frame of 2.1 kb length, coding for a Mr = 77,000 basic protein. In the aminoterminal half of the protein we detect stretches of repeated amino acids, centrally a region with homology to RNA-binding proteins containing the RNP 1 and RNP 2 consensus motif of RNA binding proteins, and the carboxyterminal part is rich in charged amino acids. The Bj6 protein is a product of the gene no-on transient A, a gene required for normal vision and courtship behaviour in Drosophila.


Role of brain imaging in disorders of brain-gut interaction: a Rome Working Team Report.
Imaging of the living human brain is a powerful tool to probe the interactions between brain, gut and microbiome in health and in disorders of brain-gut interactions, in particular IBS. While altered signals from the viscera contribute to clinical symptoms, the brain integrates these interoceptive signals with emotional, cognitive and memory related inputs in a non-linear fashion to produce symptoms. Tremendous progress has occurred in the development of new imaging techniques that look at structural, functional and metabolic properties of brain regions and networks. Standardisation in image acquisition and advances in computational approaches has made it possible to study large data sets of imaging studies, identify network properties and integrate them with non-imaging data. These approaches are beginning to generate brain signatures in IBS that share some features with those obtained in other often overlapping chronic pain disorders such as urological pelvic pain syndromes and vulvodynia, suggesting shared mechanisms. Despite this progress, the identification of preclinical vulnerability factors and outcome predictors has been slow. To overcome current obstacles, the creation of consortia and the generation of standardised multisite repositories for brain imaging and metadata from multisite studies are required.


Building waste management core indicators through Spatial Material Flow Analysis: net recovery and transport intensity indexes.
In this paper, the material and spatial characterization of the flows within a municipal solid waste (MSW) management system are combined through a Network-Based Spatial Material Flow Analysis. Using this information, two core indicators are developed for the bio-waste fraction, the Net Recovery Index (NRI) and the Transport Intensity Index (TII), which are aimed at assessing progress towards policy-related sustainable MSW management strategies and objectives. The NRI approaches the capacity of a MSW management system for converting waste into resources through a systematic metabolic approach, whereas the TII addresses efficiency in terms of the transport requirements to manage a specific waste flow throughout the entire MSW management life cycle. Therefore, both indicators could be useful in assessing key MSW management policy strategies, such as the consecution of higher recycling levels (sustainability principle) or the minimization of transport by locating treatment facilities closer to generation sources (proximity principle). To apply this methodological approach, the bio-waste management system of the region of Catalonia (Spain) has been chosen as a case study. Results show the adequacy of both indicators for identifying those points within the system with higher capacity to compromise its environmental, economic and social performance and therefore establishing clear targets for policy prioritization. Moreover, this methodological approach permits scenario building, which could be useful in assessing the outcomes of hypothetical scenarios, thus proving its adequacy for strategic planning.


Prevalence of Hypertension in Low- and Middle-Income Countries: A Systematic Review and Meta-Analysis.
We aimed to obtain overall and regional estimates of hypertension prevalence, and to examine the pattern of this disease condition across different socio-demographic characteristics in low-and middle-income countries. We searched electronic databases from inception to August 2015. We included population-based studies that reported hypertension prevalence using the current definition of blood pressure ≥140/90 mm Hg or self-reported use of antihypertensive medication. We used random-effects meta-analyses to pool prevalence estimates of hypertension, overall, by World Bank region and country income group. Meta-regression analyses were performed to explore sources of heterogeneity across the included studies. A total of 242 studies, comprising data on 1,494,609 adults from 45 countries, met our inclusion criteria. The overall prevalence of hypertension was 32.3% (95% confidence interval [CI] 29.4-35.3), with the Latin America and Caribbean region reporting the highest estimates (39.1%, 95% CI 33.1-45.2). Pooled prevalence estimate was also highest across upper middle income countries (37.8%, 95% CI 35.0-40.6) and lowest across low-income countries (23.1%, 95% CI 20.1-26.2). Prevalence estimates were significantly higher in the elderly (≥65 years) compared with younger adults (<65 years) overall and across the geographical regions; however, there was no significant sex-difference in hypertension prevalence (31.9% vs 30.8%, P = 0.6). Persons without formal education (49.0% vs 24.9%, P < 0.00001), overweight/obese (46.4% vs 26.3%, P < 0.00001), and urban settlers (32.7% vs 25.2%, P = 0.0005) were also more likely to be hypertensive, compared with those who were educated, normal weight, and rural settlers respectively. This study provides contemporary and up-to-date estimates that reflect the significant burden of hypertension in low- and middle-income countries, as well as evidence that hypertension remains a major public health issue across the various socio-demographic subgroups. On average, about 1 in 3 adults in the developing world is hypertensive. The findings of this study will be useful for the design of hypertension screening and treatment programmes in low- and middle-income countries.


Clinical spectrum of pyruvate kinase deficiency: data from the Pyruvate Kinase Deficiency Natural History Study.
An international, multicenter registry was established to collect retrospective and prospective clinical data on patients with pyruvate kinase (PK) deficiency, the most common glycolytic defect causing congenital nonspherocytic hemolytic anemia. Medical history and laboratory and radiologic data were retrospectively collected at enrollment for 254 patients with molecularly confirmed PK deficiency. Perinatal complications were common, including anemia that required transfusions, hyperbilirubinemia, hydrops, and prematurity. Nearly all newborns were treated with phototherapy (93%), and many were treated with exchange transfusions (46%). Children age 5 years and younger were often transfused until splenectomy. Splenectomy (150 [59%] of 254 patients) was associated with a median increase in hemoglobin of 1.6 g/dL and a decreased transfusion burden in 90% of patients. Predictors of a response to splenectomy included higher presplenectomy hemoglobin (P = .007), lower indirect bilirubin (P = .005), and missense PKLR mutations (P = .0017). Postsplenectomy thrombosis was reported in 11% of patients. The most frequent complications included iron overload (48%) and gallstones (45%), but other complications such as aplastic crises, osteopenia/bone fragility, extramedullary hematopoiesis, postsplenectomy sepsis, pulmonary hypertension, and leg ulcers were not uncommon. Overall, 87 (34%) of 254 patients had both a splenectomy and cholecystectomy. In those who had a splenectomy without simultaneous cholecystectomy, 48% later required a cholecystectomy. Although the risk of complications increases with severity of anemia and a genotype-phenotype relationship was observed, complications were common in all patients with PK deficiency. Diagnostic testing for PK deficiency should be considered in patients with apparent congenital hemolytic anemia and close monitoring for iron overload, gallstones, and other complications is needed regardless of baseline hemoglobin. This trial was registered at www.clinicaltrials.gov as #NCT02053480.


The Case for a Pre-Cancer Genome Atlas (PCGA).
Understanding the earliest molecular and cellular events associated with cancer initiation remains a key bottleneck to transforming our approach to cancer prevention and detection. While TCGA has provided unprecedented insights into the genomic events associated with advanced stage cancer, there have been few studies comprehensively profiling premalignant and early-stage disease or elucidating the role of the microenvironment in premalignancy and tumor initiation. In this article, we make a call for development of a "Pre-Cancer Genome Atlas (PCGA)," a concerted initiative to characterize the molecular alterations in premalignant lesions and the corresponding changes in the microenvironment associated with progression to invasive carcinoma. This initiative will require a multicenter coordinated effort to comprehensively profile (cellular and molecular) premalignant lesions and their corresponding "field of injury" collected longitudinally as the lesion progresses towards or regresses from frank malignancy across multiple tumor types. Genomic characterization of alterations in premalignant lesions and their microenvironment, for both bulk tissue and single cells, will enable development of biomarkers for early detection and risk stratification as well as allow for the development of novel targeted cancer interception strategies. The multi-institutional and multidisciplinary collaborative "big-data" effort underlying the PCGA will help usher in a new era of precision medicine for cancer detection and prevention.


An Educational Intervention to Train Professional Nurses in Promoting Patient Engagement: A Pilot Feasibility Study.
Introduction: Growing evidence recognizes that patients who are motivated to take an active role in their care can experience a range of health benefits and reduced healthcare costs. Nurses play a critical role in the effort to make patients fully engaged in their disease management. Trainings devoted to increase nurses' skills and knowledge to assess and promote patient engagement are today a medical education priority. To address this goal, we developed a program of nurse education training in patient engagement strategies (NET-PES). This paper presents pilot feasibility study and preliminary participants outcomes for NET-PES. Methods: This is a pilot feasibility study of a 2-session program on patient engagement designed to improve professional nurses' ability to engage chronic patients in their medical journey; the training mainly focused on passing patient engagement assessment skills to clinicians as a crucial mean to improve care experience. A pre-post pilot evaluation of NET-PES included 46 nurses working with chronic conditions. A course specific competence test has been developed and validated to measure patient engagement skills. The design included self-report questionnaire completed before and after the training for evaluation purposes. Participants met in a large group for didactic presentations and then they were split into small groups in which they used role-play and case discussion to reflect upon the value of patient engagement measurement in relation to difficult cases from own practice. Results: Forty-six nurses participated in the training program. The satisfaction questionnaire showed that the program met the educational objectives and was considered to be useful and relevant by the participants. Results demonstrated changes on clinicians' attitudes and skills in promoting engagement. Moreover, practitioners demonstrated increases on confidence regarding their ability to support their patients' engagement in the care process. Conclusions: Learning programs teaching nurses about patient engagement strategies and assessment measures in clinical practice are key in supporting the realization of patient engagement in healthcare. Training nurses in this area is feasible and accepted and might have an impact on their ability to engage patients in the chronic care journey. Due to the limitation of the research design, further research is needed to assess the effectiveness of such a program and to verify if the benefits envisaged in this pilot are maintained on a long-term perspective and to test results by employing a randomized control study design.


Cancer surgery among American Indians.
American Indians (AIs) have the poorest cancer survival rates of any U.S. ethnic group. Late diagnosis, poor access to specialty care, and delays in therapy likely contribute to excess mortality. Surgery plays a central role in therapy for solid organ cancer. To determine whether operative outcomes also contribute to poor long-term survival among AI patients with cancer. Population-based retrospective cohort study comparing patient- and hospital-level factors and short-term operative outcomes for AI and non-Hispanic white patients. Survey-weighted multivariate analyses assessed the effect of AI ethnicity on hospital location, in-hospital mortality, and prolonged length of stay. A 20% stratified sample of all US community hospitals. Patients undergoing oncologic resection for 1 of 20 malignant neoplasms in the Nationwide Inpatient Sample from January 1, 1998, through December 31, 2009. In-hospital mortality, length of stay, and hospital location (rural vs urban). Of 740,878 patients who met our inclusion criteria, 3048 were AIs. The AI patients were younger, more likely to undergo cancer surgery at rural hospitals, and more likely to be admitted for nonelective procedures and had more comorbidities than non-Hispanic white patients of similar ages (all, P < .05). The AI patients had comparable inpatient mortality and length of stay. CONCLUSIONS AND RELEVANCE This investigation is the largest study of surgical outcomes among AIs to date and the first to focus on cancer surgery. This relatively young cohort does not experience poor outcomes after oncologic resection. Future research should uncover other factors in the continuum of cancer care that may contribute to the poor long-term survival of AI patients with cancer, including delivery of perioperative therapies.


A one health framework to estimate the cost of antimicrobial resistance.
The costs attributable to antimicrobial resistance (AMR) remain theoretical and largely unspecified. Current figures fail to capture the full health and economic burden caused by AMR across human, animal, and environmental health; historically many studies have considered only direct costs associated with human infection from a hospital perspective, primarily from high-income countries. The Global Antimicrobial Resistance Platform for ONE-Burden Estimates (GAP-ON€) network has developed a framework to help guide AMR costing exercises in any part of the world as a first step towards more comprehensive analyses for comparing AMR interventions at the local level as well as more harmonized analyses for quantifying the full economic burden attributable to AMR at the global level. GAP-ON€ (funded under the JPIAMR 8th call (Virtual Research Institute) is composed of 19 international networks and institutions active in the field of AMR. For this project, the Network operated by means of Delphi rounds, teleconferences and face-to-face meetings. The resulting costing framework takes a bottom-up approach to incorporate all relevant costs imposed by an AMR bacterial microbe in a patient, in an animal, or in the environment up through to the societal level. The framework itemizes the epidemiological data as well as the direct and indirect cost components needed to build a realistic cost picture for AMR. While the framework lists a large number of relevant pathogens for which this framework could be used to explore the costs, the framework is sufficiently generic to facilitate the costing of other resistant pathogens, including those of other aetiologies. In order to conduct cost-effectiveness analyses to choose amongst different AMR-related interventions at local level, the costing of AMR should be done according to local epidemiological priorities and local health service norms. Yet the use of a common framework across settings allows for the results of such studies to contribute to cumulative estimates that can serve as the basis of broader policy decisions at the international level such as how to steer R&D funding and how to prioritize AMR amongst other issues. Indeed, it is only by building a realistic cost picture that we can make informed decisions on how best to tackle major health threats.


Overview of mRNA expression profiling using DNA microarrays.
DNA microarray technology allows simultaneous measurement of the mRNA levels of thousands of genes. This powerful technology has applications in addressing many biological questions that were not approachable previously; however, the enormous size of microarray data sets leads to issues of experimental design and statistical analysis that are unfamiliar to many molecular biologists. The type of array used, the design of the biological experiment, the number of experimental replicates, and the statistical method for data analysis should all be chosen based on the scientific goals of the investigator. This overview presents a discussion of the relative merits and limitations of various methods with respect to some common applications of microarray experiments.


Alteration in the venous drainage of a dural arteriovenous fistula following angioplasty.
The pattern of venous drainage from a dural arteriovenous fistula (DAVF) has been shown to affect the natural history of these lesions. Angioplasty and stent placement of the dural sinuses have been described to improve outflow in venous hypertensive states and may improve the venous drainage pattern from a DAVF. We report the case of a patient with a benign but stenosed type IIa transverse sinus DAVF who underwent angioplasty to improve venous outflow. This resulted in conversion of the DAVF to a more malignant type IIb drainage pattern with reflux into the cortical venous system.


Specific tolerance to neural allografts induced with an antibody to the interleukin 2 receptor.
Despite considerable evidence documenting the central nervous system as a site of immunological privilege, immune responses do occur within the brain and neural allografts between major histocompatibility complexes (MHC) and minor antigen incompatible rat strains may be rejected. The survival of completely MHC incompatible neural allografts has been found to be prolonged indefinitely after administration of a monoclonal antibody (mAb) to the interleukin 2 receptor (IL-2R) for 10 d after transplantation. Here we present evidence that rats with long-term surviving lateral ventricular neural allografts, after anti-IL-2R treatment, accept subsequent neural allografts from the same donor strain, placed in a peripheral nonprivileged site, but rapidly reject third-party grafts. Thus, treatment with a mAb to the p55 chain of the IL-2R has resulted in the specific acceptance of second grafts of fully allogeneic neural tissue. These results suggest that ongoing interaction between elements of the host immune system and alloantigen within the brain maintains the tolerant state and furthermore, that interruption of signaling through the IL-2R may be important in allospecific tolerance induction.


Diversity of cytosine methylation across the fungal tree of life.
The generation of thousands of fungal genomes is leading to a better understanding of genes and genomic organization within the kingdom. However, the epigenome, which includes DNA and chromatin modifications, remains poorly investigated in fungi. Large comparative studies in animals and plants have deepened our understanding of epigenomic variation, particularly of the modified base 5-methylcytosine (5mC), but taxonomic sampling of disparate groups is needed to develop unifying explanations for 5mC variation. Here, we utilize the largest phylogenetic resolution of 5mC methyltransferases (5mC MTases) and genome evolution to better understand levels and patterns of 5mC across fungi. We show that extant 5mC MTase genotypes are descendent from ancestral maintenance and de novo genotypes, whereas the 5mC MTases DIM-2 and RID are more recently derived, and that 5mC levels are correlated with 5mC MTase genotype and transposon content. Our survey also revealed that fungi lack canonical gene-body methylation, which distinguishes fungal epigenomes from certain insect and plant species. However, some fungal species possess independently derived clusters of contiguous 5mC encompassing many genes. In some cases, DNA repair pathways and the N6-methyladenine DNA modification negatively coevolved with 5mC pathways, which additionally contributed to interspecific epigenomic variation across fungi.


A comparison of electrophoretic resolution for snapshot and finish-line imaging.
Finish-line imaging, in which DNA or other macromolecules are detected after electrophoresis for a constant distance, usually improves resolution compared to snapshot imaging, in which molecules are electrophoresed for a constant time in an apparatus of comparable dimensions. Resolving power, which is an objective measure of the ability of different separatory methods to detect closely spaced molecular species, can be used to compare directly the performance of systems employing both snapshot and finish-line imaging [E. A. Ribeiro and J. C. Sutherland, Anal. Biochem. 210, 378-388 (1993)]. Experimentally determined values of resolving power are influenced both by the method of imaging (snapshot vs finish-line) and by instrument-specific factors that affect resolution. Previous comparisons of the resolving power obtained with finish-line and snapshot imaging involved data sets acquired by different instruments with different instrumental resolutions. To reduce the influence of instrumental effects, we constructed a scanning laser fluorometer that can measure both snapshot and finish-line images of fluorochrome-labeled DNA. Snapshot and finish-line images of a DNA sample containing HaeII restriction fragments of the DNA from bacteriophage T7, which range in length from 474 to 6514 base pairs, were obtained under otherwise identical electrophoretic conditions. Snapshot and finish-line imaging give similar resolving powers for DNA molecules up to about 1.5 kbp long. For both imaging modes, maximum resolving power was achieved for DNA molecules between 2 and 3 kbp in length. For larger DNA molecules, finish-line imaging provided higher resolving power. The ratio of the resolving power of finish-line images to that of snapshot images increased monotonically as a function of DNA length. For the longest restriction fragments studied (6514 bp), the resolving power for finish-line images exceeded that of snapshot images by about 50%.


Risk management in waste water treatment.
With the continuous restructuring of the water market due to liberalisation, privatisation and internationalisation processes, the requirements on waste water disposal companies have grown. Increasing competition requires a target-oriented and clearly structured procedure. At the same time it is necessary to meet the environment-relevant legal requirements and to design the processes to be environment-oriented. The implementation of risk management and the integration of such a management instrument in an existing system in addition to the use of modern technologies and procedures can help to make the operation of the waste water treatment safer and consequently strengthen market position. The risk management process consists of three phases, risk identification, risk analysis/risk assessment and risk handling, which are based on each other, as well as of the risk managing. To achieve an identification of the risks as complete as possible, a subdivision of the kind of risks (e.g. legal, financial, market, operational) is suggested. One possibility to assess risks is the portfolio method which offers clear representation. It allows a division of the risks into classes showing which areas need handling. The determination of the appropriate measures to handle a risk (e.g. avoidance, reduction, shift) is included in the concluding third phase. Different strategies can be applied here. On the one hand, the cause-oriented strategy, aiming at preventive measures which aim to reduce the probability of occurrence of a risk (e.g. creation of redundancy, systems with low susceptibility to malfunction). On the other hand, the effect-oriented strategy, aiming to minimise the level of damage in case of an undesired occurrence (e.g. use of alarm systems, insurance cover).


Identification of self-renewing myoblasts in the progeny of single human muscle satellite cells.
We demonstrate that self-renewing myoblasts can be identified in the progeny of single human muscle satellite cells (HMSC) in culture. We show, using cytoskeletal proteins and cell size as markers, that self-renewing myoblasts are phenotypically different from other myoblasts, but similar to native HMSC. Native desmin-positive HMSC, cultured as single cells, yielded two major populations of myoblasts, alpha-sarcomeric (alpha-SR)-actin-positive myoblasts and desmin-positive myoblasts. In appropriate culture conditions, alpha-SR-actin-positive myoblasts fused into myotubes, whereas a population of desmin-positive non-fusing myoblasts (NFMB) persisted for weeks among the myotubes. Upon isolation from myotubes, some of the NFMB resumed proliferation and their progeny included fusing and non-fusing myoblasts, with the same cytoskeletal phenotypes as the progeny of native HMSC. This self-renewal cycle could be repeated, yielding four cohorts of myoblasts. The yield of self-renewing cells appeared to decrease with the number of cycles. These results suggest that stem cells are present among NFMB. Moreover, we find that these presumptive stem cells are already segregated during myoblast proliferation. They are small, phenotypically similar to native HMSC, and do not divide unless they are isolated from their sister progeny and cultured alone. Enriched preparations of cells with stem cell-like properties can be obtained from proliferating myoblasts by flow cytometry on the basis of size and nucleocytoplasmic ratio.


Quality of life measures predict cardiovascular health and physical performance in chronic renal failure patients.
Patients with advanced chronic kidney disease (CKD) experience complex functional and structural changes of the cardiopulmonary and musculoskeletal system. This results in reduced exercise tolerance, quality of life and ultimately premature death. We investigated the relationship between subjective measures of health related quality of life and objective, standardised functional measures for cardiovascular and pulmonary health. Between April 2010 and January 2013, 143 CKD stage-5 or CKD5d patients (age 46.0±1.1y, 62.2% male), were recruited prospectively. A control group of 83 healthy individuals treated for essential hypertension (HTN; age 53.2±0.9y, 48.22% male) were recruited at random. All patients completed the SF-36 health survey questionnaire, echocardiography, vascular tonometry and cardiopulmonary exercise testing. Patients with CKD had significantly lower SF-36 scores than the HTN group; for physical component score (PCS; 45.0 vs 53.9, p<0.001) and mental component score (MCS; 46.9 vs. 54.9, p<0.001). CKD subjects had significantly poorer exercise tolerance and cardiorespiratory performance compared with HTN (maximal oxygen uptake; VO2peak 19.9 vs 25.0ml/kg/min, p<0.001). VO2peak was a significant independent predictor of PCS in both groups (CKD: b = 0.35, p = 0.02 vs HTN: b = 0.27, p = 0.001). No associations were noted between PCS scores and echocardiographic characteristics, vascular elasticity and cardiac biomarkers in either group. No associations were noted between MCS and any variable. The interaction effect of study group with VO2peak on PCS was not significant (ΔB = 0.08; 95%CI -0.28-0.45, p = 0.7). However, overall for a given VO2peak, the measured PCS was much lower for patients with CKD than for HTN cohort, a likely consequence of systemic uremia effects. In CKD and HTN, objective physical performance has a significant effect on quality of life; particularly self-reported physical health and functioning. Therefore, these quality of life measures are indeed a good reflection of physical health correlating highly with objective physical performance measures.


Evaluation of disease severity with quantitative chest CT in COVID-19 patients.
We aimed to assess the severity of coronavirus disease 2019 (COVID-19) pneumonia on computed tomography (CT) using quantitative (QCT) and semiquantitative (SCT) assessments and compare with the clinical findings. Two observers independently examined the CT images of COVID-19 patients, and the SCT severity score was calculated. The SCT score was calculated as the sum of values ranging from 0 to 4, according to the volumetric rate of involvement for each lung lobe. In quantitative assessment, total lung volume (TLV) was automatically calculated from CT density values between -200 and -950 HU. Besides, healthy lung volume (HLV) was calculated from voxels between -800 and -950 HU. The QCT score was calculated with the following formula: (TLV - HLV / TLV) ×100. All patients were clinically divided into four groups: mild, common, severe, and critical. Interobserver agreement for SCT assessment was investigated using the Cohen's Kappa statistics (κ). Pearson's correlation coefficient was used for the relationship between continuous data. The diagnostic accuracy of SCT and QCT in the differentiation of clinically limited (mild, common) and extensive (severe, critical) disease was investigated using ROC analysis. Seventy-six patients with a diagnosis of COVID-19 were included. There was good agreement between the two observers in the SCT evaluation of pulmonary disease severity (κ = 0.796; 95% CI, 0.751-0.841). A significant correlation was found between QCT and SCT scores (p < 0.001, r = 0.661). Both QCT and SCT scores showed a significant correlation with clinical severity score (p < 0.001, r = 0.620 and p = 0.004, r = 0.529, respectively). The ROC analysis revealed the AUC of QCT and SCT for differentiation of limited and extensive disease as 0.873 (95% CI, 0.774-0.972) and 0.816 (95% CI, 0.673-0.959), respectively. The QCT assessment is an objective method in the evaluation of COVID-19 severity and is more successful than semiquantitative CT assessment to discriminate extensive from limited disease.


Analysis of an Unusual Case of Nonfluent Aphasia With Predominantly Posterior Perisylvian Lesion: An Apparent Paradox.
Lesion site-aphasia type discordance has garnered increasing interest in cognitive neuroscience over the last century. Diaschisis, the network concept of cognitive functions, and interindividual variability are among the plausible explanations cited in the literature for such unusual clinical cases. We describe here the case of a nonfluent type of aphasia following an ischemic stroke predominantly affecting the left posterior perisylvian cortex in a right-handed Bengali-speaking woman. The patient's comprehension was well preserved; however, she presented with a severe motor speech defect. MRI revealed an ischemic lesion in the left parietotemporal area, with slight involvement of the postero-inferior frontal cortex. We suggest two plausible explanations for this lesion-aphasia discordance: Our patient had bilateral representation of language receptive functions in her brain, and additional areas neighboring the classical Broca area may support some critical mechanisms of speech production. Taken together, these explanations may clarify why our patient was able to maintain the ability to decode language even though her language production was significantly affected.


The effects of topical and systemic Beta glucan administration on wound healing impaired by corticosteroids.
Background. Corticosteroid hormones are widely used to treat a variety of diseases. Corticosteriods have been shown to impair wound healing, which has become a serious clinical problem in wound care. The present study was designed to evaluate the efficacy of topical and systemic beta glucan administration on wound healing impaired by corticosteroids. Wistar albino rats were used for the incision and excision wound models. Percentage of wound contraction, epithelialization period, hydroxyproline level, histopathological examination, and tensile strength were evaluated. Although both systemic and local administration of beta glucan enhanced percentage wound contraction, improved epithelialization time, tensile strength, and elevated hydroxyproline level, systemic administration was found to be more effective. These results indicate that systemic and topical beta glucan improve wound healing that has been impaired by corticosteroids, and that systemic administration is more effective than topical application.


Neutralising antibodies for West Nile virus in horses from Brazilian Pantanal.
Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for WNV and its most closely-related flaviviruses that circulate in Brazil to confirm the detection of specific virus-neutralising antibodies. Of the 93 (55.4%) blocking ELISA-seropositive horse serum samples, five (3%) were seropositive for WNV, nine (5.4%) were seropositive for St. Louis encephalitis virus, 18 (10.7%) were seropositive for Ilheus virus, three (1.8%) were seropositive for Cacipacore virus and none were seropositive for Rocio virus using PRNT90, with a criteria of ≥ four-fold antibody titre difference. All caimans were negative for flaviviruses-specific antibodies using the blocking ELISA. No virus genome was detected from caiman blood or mosquito samples. The present study is the first report of confirmed serological evidence of WNV activity in Brazil.


Continental-scale tree-ring-based projection of Douglas-fir growth: Testing the limits of space-for-time substitution.
A central challenge in global change research is the projection of the future behavior of a system based upon past observations. Tree-ring data have been used increasingly over the last decade to project tree growth and forest ecosystem vulnerability under future climate conditions. But how can the response of tree growth to past climate variation predict the future, when the future does not look like the past? Space-for-time substitution (SFTS) is one way to overcome the problem of extrapolation: the response at a given location in a warmer future is assumed to follow the response at a warmer location today. Here we evaluated an SFTS approach to projecting future growth of Douglas-fir (Pseudotsuga menziesii), a species that occupies an exceptionally large environmental space in North America. We fit a hierarchical mixed-effects model to capture ring-width variability in response to spatial and temporal variation in climate. We found opposing gradients for productivity and climate sensitivity with highest growth rates and weakest response to interannual climate variation in the mesic coastal part of Douglas-fir's range; narrower rings and stronger climate sensitivity occurred across the semi-arid interior. Ring-width response to spatial versus temporal temperature variation was opposite in sign, suggesting that spatial variation in productivity, caused by local adaptation and other slow processes, cannot be used to anticipate changes in productivity caused by rapid climate change. We thus substituted only climate sensitivities when projecting future tree growth. Growth declines were projected across much of Douglas-fir's distribution, with largest relative decreases in the semiarid U.S. Interior West and smallest in the mesic Pacific Northwest. We further highlight the strengths of mixed-effects modeling for reviving a conceptual cornerstone of dendroecology, Cook's 1987 aggregate growth model, and the great potential to use tree-ring networks and results as a calibration target for next-generation vegetation models.


Suppression of HBV by tenofovir in HBV/HIV coinfected patients: a systematic review and meta-analysis.
Hepatitis B coinfection is common in HIV-positive individuals and as antiretroviral therapy has made death due to AIDS less common, hepatitis has become increasingly important. Several drugs are available to treat hepatitis B. The most potent and the one with the lowest risk of resistance appears to be tenofovir (TDF). However there are several questions that remain unanswered regarding the use of TDF, including the proportion of patients that achieves suppression of HBV viral load and over what time, whether suppression is durable and whether prior treatment with other HBV-active drugs such as lamivudine, compromises the efficacy of TDF due to possible selection of resistant HBV strains. A systematic review and meta-analysis following PRISMA guidelines and using multilevel mixed effects logistic regression, stratified by prior and/or concomitant use of lamivudine and/or emtricitabine. Data was available from 23 studies including 550 HBV/HIV coinfected patients treated with TDF. Follow up was for up to seven years but to ensure sufficient power the data analyses were limited to three years. The overall proportion achieving suppression of HBV replication was 57.4%, 79.0% and 85.6% at one, two and three years, respectively. No effect of prior or concomitant 3TC/FTC was shown. Virological rebound on TDF treatment was rare. TDF suppresses HBV to undetectable levels in the majority of HBV/HIV coinfected patients with the proportion fully suppressed continuing to increase during continuous treatment. Prior treatment with 3TC/FTC does not compromise efficacy of TDF treatment. The use of combination treatment with 3TC/FTC offers no significant benefit over TDF alone.


The influence of hair-removal methods on wound infections.
The influence of preoperative shaving v clipping on wound infection rate was studied in 1,013 patients undergoing elective operations at a single hospital. Patients were prospectively randomized to be either shaved or clipped the night before or the morning of operation. The AM clipper method was associated with significantly fewer infections than were the other methods, both at discharge and at 30-day follow-up. The greatest benefit was in the group with clean wounds. For each 1,000 patients treated, a savings of approximately $270,000 could be realized if the AM clipper method replaced shaving for preoperative hair removal. Preoperative shaving is deleterious, and the practice should be abandoned.


Anti-nerve growth factor does not change physical activity in normal young or aging mice but does increase activity in mice with skeletal pain.
Anti-nerve growth factor (anti-NGF) therapy has shown significant promise in attenuating several types of skeletal pain. However, whether anti-NGF therapy changes the level of physical activity in individuals with or without skeletal pain is largely unknown. Here, automated day/night activity boxes monitored the effects of anti-NGF treatment on physical activity in normal young (3 months old) and aging (18-23 months old) mice and mice with bone fracture pain. Although aging mice were clearly less active and showed loss of bone mass compared with young mice, anti-NGF treatment had no effect on any measure of day/night activity in either the young or aging mice. By contrast, in mice with femoral fracture pain, anti-NGF treatment produced a clear increase (10%-27%) in horizontal activity, vertical rearing, and velocity of travel compared with the Fracture + Vehicle group. These results suggest, just as in humans, mice titrate their level of physical activity to their level of skeletal pain. The level of skeletal pain may in part be determined by the level of free NGF that seems to rise after injury but not normal aging of the skeleton. In terms of bone healing, animals that received anti-NGF showed an increase in the size of calcified callus but no increase in the number of displaced fractures or time to cortical union. As physical activity is the best nondrug treatment for many patients with skeletal pain, anti-NGF may be useful in reducing pain and promoting activity in these patients.


Effect of vascular endothelial growth factor C (VEGF-C) gene transfer in rat model of secondary lymphedema.
Secondary lymphedema has been clinically well described, but a cure is still lacking. Although there have been previous investigations using plasmid DNA for gene therapy, few have focused on the use for the treatment of lymphedema. Therefore, we investigated the effects of VEGF-C gene transfer for the treatment of lymphedema using our plasmid pcDNA3.1-VEGF-C. We produced a surgical model of secondary lymphedema in the rat hindlimb and treated with local intradermal VEGF-C transfection to investigate the efficacy of gene transfer. Magnetic resonance imaging (MRI) (P<0.05), B ultrasound (P<0.05), and water displacement volumetry (P<0.05) demonstrated a reduction of lymphedema in therapy group as compared to controls. Histological and immunofluorescent studies demonstrated numerous newly formed lymphatic vessels in therapy group. Our results indicate that VEGF-C gene therapy has produced new lymphatic vessels which may have improved functional lymphatic drainage to reduce lymphedema volume in our model.


Prospective longitudinal study of signs and symptoms associated with primary tooth eruption.
To assess the association between primary tooth eruption and the manifestation of signs and symptoms of teething in infants. An 8-month, longitudinal study was conducted with 47 noninstitutionalized infants (ie, receiving care at home) between 5 and 15 months of age in the city of Diamantina, Brazil. The nonrandomized convenience sample was based on the registry of infants in this age range provided by the Diamantina Secretary of Health. Eligible participants were infants with up to 7 erupted incisors and no history of chronic disease or disorders that could cause an increase in the signs and symptoms assessed in the study. Tympanic and axillary temperature readings and clinical oral examinations were performed daily. A daily interview with the mothers was conducted to investigate the occurrence of 13 signs and symptoms associated with teething presented by the infants in the previous 24 hours. Teething was associated with a rise in tympanic temperature on the day of the eruption (P = .004) and with the occurrence of other signs and symptoms. Readings of maximal tympanic and axillary temperatures were 36.8°C and 36.6°C, respectively. The most frequent signs and symptoms associated with teething were irritability (median: 0.60; P < .001), increased salivation (median: 0.50; P < .001), runny nose (median: 0.50; P < .001), and loss of appetite (median: 0.50; P < .001). Irritability, increased salivation, runny nose, loss of appetite, diarrhea, rash, and sleep disturbance were associated with primary tooth eruption. Results of this study support the concept that the occurrence of severe signs and symptoms, such as fever, could not be attributed to teething.


Geometric control of cell life and death.
Human and bovine capillary endothelial cells were switched from growth to apoptosis by using micropatterned substrates that contained extracellular matrix-coated adhesive islands of decreasing size to progressively restrict cell extension. Cell spreading also was varied while maintaining the total cell-matrix contact area constant by changing the spacing between multiple focal adhesion-sized islands. Cell shape was found to govern whether individual cells grow or die, regardless of the type of matrix protein or antibody to integrin used to mediate adhesion. Local geometric control of cell growth and viability may therefore represent a fundamental mechanism for developmental regulation within the tissue microenvironment.


Mechanism to trigger unfolding in O(6) -alkylguanine-DNA alkyltransferase.
O(6) -alkylguanine-DNA alkyltransferase (AGT) adopts a non-enzymatic suicide mechanism for the repair of methylated guanine bases by transferring the methyl adduct to itself, thereby initiating unfolding and fast degradation. Classical molecular dynamics simulations provide quantitative evidence that two conserved glycine residues at the centre of an α-helix make the structure susceptible to structural perturbations. The stability of this helix, designated the "recognition helix", is an important factor during the early onset of unfolding of human AGT (hAGT). By combining theory and experiment, we found that helical stability is controlled by key factors in the surrounding protein structure. By using a "double-clip" mechanism, nearby residues hydrogen bond to both the base and centre of the helix. This double clip stabilises this site in the protein in the absence of substrate, but the helix is destabilised upon alkylation. The present investigation aimed to establish why alkylation of hAGT leads to conformational changes and how the protein environment functions as a switch, thus turning the stability of the protein "on" or "off" to tune degradability.


5-Carboxylcytosine is localized to euchromatic regions in the nuclei of follicular cells in axolotl ovary.
5-Methylcytosine (5-mC) is an epigenetic modification associated with gene repression. Recent studies demonstrated that 5-mC can be enzymatically oxidised into 5-hydroxymethylcytosine and further into 5-formylcytosine (5-fC) and 5-carboxylcytsine (5-caC). 5-caC has been found in embryonic stem cells and in mouse pre-implantation embryos but no detectable levels of this modification have been reported for somatic tissues to date. Whereas it has been suggested that 5-caC can serve as an intermediate in the process of active demethylation, the function of this form of modified cytosine remains obscure. Here we show that 5-caC is immunochemically detectable in somatic cells of axolotl ovary. We demonstrate that both 5-hmC and 5-caC are localized to the euchromatin in the nuclei of axolotl follicular cells with similar patterns of spatial distribution. Our results suggest that 5-carboxylcytosine may play a distinct functional role in certain biological contexts.


Identification and characterization of a human tRNA-specific adenosine deaminase related to the ADAR family of pre-mRNA editing enzymes.
The mammalian adenosine deaminases acting on RNA (ADARs) constitute a family of sequence-related proteins involved in pre-mRNA editing of nuclear transcripts through site-specific adenosine modification. We report here the identification and characterization of a human ADAR protein, hADAT1, that specifically deaminates adenosine 37 to inosine in eukaryotic tRNA(Ala). It represents the functional homologue of the recently identified yeast protein Tad1p [Gerber, A., Grosjean, H., Melcher, T. & Keller, W. (1998) EMBO J. 17, 4780-4789]. The hADAT1 cDNA predicts a protein of 502 aa whose sequence displays strongest overall homology to a Drosophila melanogaster ORF (50% similarity, 32% identity), and the catalytic domain is closely related to the other ADAR proteins. In vitro, the recombinantly expressed and purified hADAT1 protein efficiently and specifically deaminates A(37) in the anticodon loop of tRNA(Ala) from higher eukaryotes and with lower efficiency from lower eukaryotes. It does not modify adenosines residing in double-stranded RNA or in pre-mRNAs that serve as substrates for ADAR1 or ADAR2. The anticodon stem-loop of tRNA(Ala) alone is not a functional substrate for hADAT1. The enzyme is expressed ubiquitously in human tissues and is represented by a single gene. The identification and cloning of hADAT1 should help to elucidate the physiological significance of this unique modification in tRNA(Ala), which is conserved from yeast to man.


A 50-year-old man with chronic low back pain.
Mr S, a 50-year-old man, has long-standing low back pain. His pain began more than 20 years earlier with a lumbar disk herniation and has persisted despite diskectomy. He has undergone numerous treatments, but he remains disabled with ongoing pain. His treatment course is used to frame the epidemiology and pathophysiology underlying acute and chronic lumbosacral and radicular pain. The roles of neuropathic pain medications, chronic opioid therapy, physical therapy, spinal manipulation, and multidisciplinary pain treatment programs are reviewed. The indications for and outcomes associated with interventional pain treatments, including epidural steroid injection, facet blocks and radiofrequency treatment for facet-related pain, intradiskal electrothermal therapy, spinal cord stimulation, and intrathecal drug delivery, are discussed. Clinicians are given an evidence-based approach to using available treatment options for low back pain.


Answering Wicked Questions: Dealing With Opposing Truths as a Nursing Associate Professor.
Nursing associate professors frequently are confronted with increasing responsibilities and fewer resources. These challenges commonly contribute to declines in job satisfaction and may result in departing academe. This article addresses these challenges by providing answers to four common "wicked questions" experienced by nursing associate professors: (a) How do I decline a request from a supervisor to take on additional responsibilities while continuing to support the mission of the school and advance my own scholarly productivity? (b) How do I handle the workload of multiple doctoral students with a variety of content areas that are different from my own and maintain my own level of productivity? (c) How do I handle expectations for more service, and leadership for the school, university, and professional organizations, yet the teaching and research responsibilities have not changed or have increased? and (d) What are some additional tips to being a more productive nursing associate professor?


Clinical implications of pleomorphic ventricular tachycardias on oral sotalol therapy.
In 90 consecutive patients with coronary artery disease and sustained monomorphic ventricular tachycardia, who were treated with oral sotalol and underwent programmed stimulation to determine drug effectiveness, the influence of sotalol on induced ventricular tachycardia morphology was retrospectively examined. In 54 patients (60%) sotalol rendered the tachycardia non-inducible. However, contrary to drug-testing with class I antiarrhythmic agents, induction of multiple morphologies at baseline study did not predict failure of subsequent drug-testing with sotalol. In the remaining 36 patients (40%), in whom sotalol did not modify inducibility, 21 patients (i.e. a total of 23%) manifested at least one new morphology during electropharmacological testing on sotalol. This effect was independent of the degree of left ventricular dysfunction, infarct location and numbers of morphologies at baseline, but corresponded with drug-induced changes in refractoriness. This observation may be related to a proarrhythmic effect of sotalol. Slowing of ventricular tachycardia rate and changes in morphology may have implications in patients receiving implantable cardioverter-defibrillators or those undergoing ablative procedures.


Association between lymph node evaluation for colon cancer and node positivity over the past 20 years.
Among patients surgically treated for colon cancer, better survival has been demonstrated in those with more lymph nodes evaluated. The presumed mechanism behind this association suggests that a more extensive lymph node evaluation reduces the risk of understaging, leading to improved survival. To further evaluate the mechanism behind lymph node evaluation and survival by examining the association between more extensive lymph node evaluation, identification of lymph node-positive cancers, and hazard of death. Observational cohort study. Surveillance, Epidemiology, and End Results (SEER) program data from 1988 through 2008. 86,394 patients surgically treated for colon cancer. We examined the relationship between lymph node evaluation and node positivity using Cochran-Armitage tests and multivariate logistic regression. The association between lymph node evaluation and hazard of death was evaluated using Cox proportional hazards modeling. The number of lymph nodes evaluated increased from 1988 to 2008 but did not result in a significant overall increase in lymph node positivity. During 1988-1990, 34.6% of patients (3875/11,200) had 12 or more lymph nodes evaluated, increasing to 73.6% (9798/13,310) during 2006-2008 (P < .001); however, the proportion of node-positive cancers did not change with time (40% in 1988-1990, 42% in 2006-2008, P = .53). Although patients with high levels of lymph node evaluation were only slightly more likely to be node positive (adjusted odds ratio for 30-39 nodes vs 1-8 nodes, 1.11; 95% CI, 1.02-1.20), these patients experienced significantly lower hazard of death compared with those with fewer nodes evaluated (adjusted hazard ratio for 30-39 nodes vs 1-8 nodes, 0.66; 95% CI, 0.62-0.71; unadjusted 5-year mortality, 35.3%). The number of lymph nodes evaluated for colon cancer has markedly increased in the past 2 decades but was not associated with an overall shift toward higher-staged cancers, questioning the upstaging mechanism as the primary basis for improved survival in patients with more lymph nodes evaluated.


Accumulation of p53 protein is frequent in ovarian cancers associated with BRCA1 and BRCA2 germline mutations.
Mutations in the BRCA1 or BRCA2 genes are responsible for up to 95% of hereditary ovarian cancer cases. Both genes function as tumour suppressor genes, and development of a cancer is thought to require an accumulation of somatic genetic events in addition to the inherited germline predisposition. It is unknown whether these somatic events in BRCA associated ovarian cancer are similar to or distinct from those in sporadic cases. The most frequent somatic genetic event in ovarian cancer is a mutation of the p53 gene. To study the role of p53 in hereditary ovarian cancer, by analysing accumulation of the p53 protein in ovarian cancers which occurred in BRCA1 or BRCA2 germline mutation carriers and comparing the results with a panel of ovarian cancers from patients who tested negative for both BRCA1 and BRCA2. The study group consisted of 39 ovarian cancer patients in whom a BRCA mutation had been confirmed previously. p53 Immunohistochemistry was performed on archival tissue using a standard microwave antigen retrieval technique. The rate of p53 accumulation was compared with 40 ovarian cancer cases who tested negative for BRCA1 and BRCA2 germline mutations. P53 Accumulation was similar in BRCA related ovarian cancers and BRCA negative controls. Overall 27 of 39 BRCA1 or BRCA2 positive cases (69%) had evidence of p53 accumulation, compared with 24 of 40 invasive ovarian cancer cases (60%) which tested negative for BRCA1 and BRCA2 germline mutations. BRCA1 related ovarian cancers showed p53 accumulation in 22 of 30 cases (73%); p53 accumulation was present in five of nine BRCA2 related ovarian cancers. In addition to germline BRCA1 and BRCA2 mutations, somatic p53 alterations leading to p53 accumulation are an important event in hereditary ovarian cancer and are as frequent as in non-BRCA-related ovarian cancer.


High-density genome-wide association study for residual feed intake in Holstein dairy cattle.
Improving feed efficiency (FE) of dairy cattle may boost farm profitability and reduce the environmental footprint of the dairy industry. Residual feed intake (RFI), a candidate FE trait in dairy cattle, can be defined to be genetically uncorrelated with major energy sink traits (e.g., milk production, body weight) by including genomic predicted transmitting ability of such traits in genetic analyses for RFI. We examined the genetic basis of RFI through genome-wide association (GWA) analyses and post-GWA enrichment analyses and identified candidate genes and biological pathways associated with RFI in dairy cattle. Data were collected from 4,823 lactations of 3,947 Holstein cows in 9 research herds in the United States. Of these cows, 3,555 were genotyped and were imputed to a high-density list of 312,614 SNP. We used a single-step GWA method to combine information from genotyped and nongenotyped animals with phenotypes as well as their ancestors' information. The estimated genomic breeding values from a single-step genomic BLUP were back-solved to obtain the individual SNP effects for RFI. The proportion of genetic variance explained by each 5-SNP sliding window was also calculated for RFI. Our GWA analyses suggested that RFI is a highly polygenic trait regulated by many genes with small effects. The closest genes to the top SNP and sliding windows were associated with dry matter intake (DMI), RFI, energy homeostasis and energy balance regulation, digestion and metabolism of carbohydrates and proteins, immune regulation, leptin signaling, mitochondrial ATP activities, rumen development, skeletal muscle development, and spermatogenesis. The region of 40.7 to 41.5 Mb on BTA25 (UMD3.1 reference genome) was the top associated region for RFI. The closest genes to this region, CARD11 and EIF3B, were previously shown to be related to RFI of dairy cattle and FE of broilers, respectively. Another candidate region, 57.7 to 58.2 Mb on BTA18, which is associated with DMI and leptin signaling, was also associated with RFI in this study. Post-GWA enrichment analyses used a sum-based marker-set test based on 4 public annotation databases: Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, Reactome pathways, and medical subject heading (MeSH) terms. Results of these analyses were consistent with those from the top GWA signals. Across the 4 databases, GWA signals for RFI were highly enriched in the biosynthesis and metabolism of amino acids and proteins, digestion and metabolism of carbohydrates, skeletal development, mitochondrial electron transport, immunity, rumen bacteria activities, and sperm motility. Our findings offer novel insight into the genetic basis of RFI and identify candidate regions and biological pathways associated with RFI in dairy cattle.


Germline mutations in hereditary diffuse gastric cancer.
Gastric cancer is one of the leading causes of cancer-related deaths worldwide. Among which, about 1%-3% of gastric cancer patients were characterized by inherited gastric cancer predisposition syndromes, knowing as hereditary diffuse gastric cancer (HDGC). Studies reported that CDH1 germline mutations are the main cause of HDGC. With the help of rapid development of genetic testing technologies and data analysis tools, more and more researchers focus on seeking candidate susceptibility genes for hereditary cancer syndromes. In addition, National Comprehensive Cancer Network (NCCN) guidelines recommend that the patients of HDGC carrying CDH1 mutations should undergo prophylactic gastrectomy or routine endoscopic surveillances. Therefore, genetic counseling plays a key role in helping individuals with pathogenic mutations make appropriate risk management plans. Moreover, experienced and professional genetic counselors as well as a systematic multidisciplinary team (MDT) are also required to facilitate the development of genetic counseling and benefit pathogenic mutation carriers who are in need of regular and standardized risk management solutions. In this review, we provided an overview about the germline mutations of several genes identified in HDGC, suggesting that these genes may potentially act as susceptibility genes for this malignant cancer syndrome. Furthermore, we introduced information for prevention, diagnosis and risk management of HDGC. Investigations on key factors that may have effect on risk management decision-making and genetic data collection of more cancer syndrome family pedigrees are required for the development of HDGC therapeutic strategies.


Elucidation of the molecular basis of cholecystokinin Peptide docking to its receptor using site-specific intrinsic photoaffinity labeling and molecular modeling.
G protein-coupled receptors represent the largest family of receptors and the major target of current drug development efforts. Understanding of the mechanisms of ligand binding and activation of these receptors remains limited, despite recent advances in structural determination of family members. This work focuses on the use of photoaffinity labeling and molecular modeling to elucidate the structural basis of binding a natural peptide ligand to a family A G protein-coupled receptor, the type 1 cholecystokinin receptor. Two photolabile cholecystokinin analogues were developed and characterized as representing high-affinity, fully biologically active probes with sites of covalent attachment at positions 28 and 31. The sites of receptor labeling were identified by purification, proteolytic peptide mapping, and radiochemical sequencing of labeled wild-type and mutant cholecystokinin receptors. The position 28 probe labeled second extracellular loop residue Leu(199), while the position 31 probe labeled first extracellular loop residue Phe(107). Along with five additional spatial approximation constraints coming from previous photoaffinity labeling studies and 12 distance restraints from fluorescence resonance energy transfer studies, these were built into two homology models of the cholecystokinin receptor, based on the recent crystal structures of the beta2-adrenergic receptor and A2a-adenosine receptor. The resultant agonist ligand-occupied receptor models fully accommodate all existing experimental data and represent the best refined models of a peptide hormone receptor in this important family.


The bowfin genome illuminates the developmental evolution of ray-finned fishes.
The bowfin (Amia calva) is a ray-finned fish that possesses a unique suite of ancestral and derived phenotypes, which are key to understanding vertebrate evolution. The phylogenetic position of bowfin as a representative of neopterygian fishes, its archetypical body plan and its unduplicated and slowly evolving genome make bowfin a central species for the genomic exploration of ray-finned fishes. Here we present a chromosome-level genome assembly for bowfin that enables gene-order analyses, settling long-debated neopterygian phylogenetic relationships. We examine chromatin accessibility and gene expression through bowfin development to investigate the evolution of immune, scale, respiratory and fin skeletal systems and identify hundreds of gene-regulatory loci conserved across vertebrates. These resources connect developmental evolution among bony fishes, further highlighting the bowfin's importance for illuminating vertebrate biology and diversity in the genomic era.


Enhanced Fcgamma receptor I, alphaMbeta2 integrin receptor expression by monocytes and neutrophils in rheumatoid arthritis: interaction with platelets.
To investigate platelet and leukocyte activation and interaction in patients with rheumatoid arthritis (RA) and the effect of methotrexate (MTX) or anti-tumor necrosis factor-a (TNF-a) treatment on these variables. Four-color flow cytometry analysis was performed for quantitative measurement of platelet (P-selectin, PAC-1) and leukocyte (CD11b, CD64) activation markers and estimation of percentage of leukocyte-platelet complexes in whole blood in 20 patients with RA before and after 6 weeks of therapy and in 20 controls. In addition, measures of soluble P-selectin (sP-selectin), beta-thromboglobulin, fibrinogen, prothrombin fragment 1+2, D-dimer, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), interleukin 6 (IL-6), and TNF-a and tender and swollen joint counts were carried out. Before therapy, PAC-1 binding, expression of CD11b and CD64 on monocytes and neutrophils, circulating levels of monocyte (CD11b+ or CD64+)-platelet complexes, monocyte-PAC-1+ platelet complexes, CRP, ESR, IL-6, TNF-a, fibrinogen, D-dimer and sP-selectin were significantly higher in RA patients compared to controls. The anti-TNF-a therapy significantly reduced levels of monocyte-PAC-1+ platelet complexes, sP-selectin, CRP, ESR, IL-6, TNF-a, fibrinogen, and D-dimer and tender and swollen joint counts. CD64 expression on monocytes was significantly decreased by MTX therapy. PAC-1 binding was not inhibited by MTX or anti-TNF-a. Increased platelet and leukocyte activation and increased formation of leukocyte-platelet complexes in patients with RA suggest a status of simultaneous activation of the immune and hemostatic systems.


New treatments for levodopa-induced motor complications.
Levodopa (l-dopa)-induced motor complications, including motor fluctuations and dyskinesia, affect almost all patients with Parkinson's disease (PD) at some point during the disease course, with relevant implications in global health status. Various dopaminergic and nondopaminergic pharmacological approaches as well as more invasive strategies including devices and functional surgery are available to manage such complications. In spite of undisputable improvements during the last decades, many patients remain significantly disabled, and a fully satisfying management of l-dopa-induced motor complications is still an important unmet need of PD therapy. This article reviews the recent trial results published from 2013 to April 2015 about pharmacological and nonpharmacological interventions to treat motor complications. Randomized controlled trials conducted in patients suffering from already established complications showed that new levodopa (l-dopa) formulations such as intrajejunal l-dopa-carbidopa infusion and bilayered extended-release l-dopa-carbidopa (IPX066) can improve motor fluctuations. Positive results were also obtained with a new monoamine oxidase B (MAO-B) inhibitor (safinamide) and a catechol-O-methyltransferase COMT inhibitor (opicapone). Pilot data suggest that new formulations of dopamine agonists (inhaled apomorphine) are also of potential interest. The development of novel nondopaminergic adenosine A2A antagonists (istradefylline, preladenant, and tozadenant) to treat motor fluctuations showed conflicting results in phase 2 and phase 3 trials. For dyskinesia, trials with new amantadine extended-release formulations confirmed the interest of the glutamatergic N-methyl-d-aspartate (NMDA) antagonist approach. Positive pilot antidyskinetic effects were also recently reported using serotonin agents such as eltoprazine and glutamate mGluR5 modulators such as mavoglurant. However, the translation to clinical practice of such innovative concepts remains challenging, because subsequent phase 2 trials conducted to confirm the antidyskynetic effects of mavoglurant failed, leading to the interruption of the development of this compound for this indication.