Carotid sinus baroreceptor reflex in dogs with experimental heart failure.
We have previously demonstrated a decrease in baroreceptor discharge sensitivity in dogs with experimental heart failure. In the present study, we determined the sensitivity of the carotid sinus baroreceptor reflex in dogs with pacing-induced heart failure. The carotid sinus baroreceptor reflex sensitivity was determined by pressurizing one carotid sinus with all other baroreceptor and cardiopulmonary receptor inputs removed. The data were analyzed by plotting carotid sinus pressure-mean arterial pressure curves and carotid sinus pressure-renal sympathetic nerve activity curves in the two groups of dogs. The peak arterial pressure during carotid hypotension was significantly depressed in dogs with heart failure compared with normal dogs (107.1 +/- 5.7 versus 139.8 +/- 7.0 mm Hg, p less than 0.001). Mean arterial pressure range, renal sympathetic nerve activity range, and peak slope were significantly decreased in the heart-failure group. To determine if this depression was completely due to depression of baroreceptor discharge per se, or to alterations in central or end-organ responsiveness, similar experiments were performed by stimulating the carotid sinus nerve and evaluating frequency, voltage, and duration of stimulation on the resultant mean arterial pressure and renal sympathetic nerve activity. As was the case with carotid sinus pressurization, electrical stimulation caused a significantly smaller change in mean arterial pressure in heart-failure dogs compared with the normal dogs. However, there was no significant difference between normal and heart-failure dogs for the renal sympathetic nerve activity-electrical stimulation curves. These data strongly suggest that the depressed carotid sinus baroreceptor reflex in heart failure is not solely the result of depressed baroreceptor responsiveness but may be related to poor end-organ responses and normal central control of renal sympathetic outflow.


Progress in the biotechnology of trees.
An increasing world population and rise in demand for tree products, especially wood, has increased the need to produce more timber through planting more forest with improved quality stock. Superior trees are likely to arise from several sources. Firstly, forest trees can be selected from wild populations and cloned using macropropagation techniques already being investigated for fruit tree rootstocks. Alternatively, propagation might be brought aboutin vitro through micropropagation or sustained somatic embryogenesis, with encapsulation of the somatic embryos to form artificial seeds. Tree quality could be improved through increased plant breeding and it is likely that experienced gained, to date, in the breeding of fruit species will be useful in devising strategies for forest trees. Since the development of techniques to regenerate woody plants from explant tissues, cells and protoplasts, it is now feasible to test the use of tissue culture methods to bring about improvements in tree quality. Success has already been achieved for tree species in the generation of somaclonal and protoclonal variation, the formation of haploids, triploids and polyploids, somatic hybrids and cybrids and the introduction of foreign DNA through transformation. This review summarizes the advances made so far in tree biotechnology, and suggests some of the directions that it might take in the future.


Developing the evidentiary basis for family medicine in the global context: The Besrour Papers: a series on the state of family medicine in the world.
To provide an overview of the main methodologic challenges to finding definitive evidence of the positive effects of family medicine and family medicine training on a global scale. In 2012, 2013, and 2014, the College of Family Physicians of Canada hosted the Besrour Conferences to reflect on its role in advancing the discipline of family medicine globally. The Besrour Papers Working Group, which was struck at the 2013 conference, was tasked with developing a series of papers to highlight the key issues, lessons learned, and outcomes emerging from the various activities of the Besrour collaboration. The working group comprised members of various academic departments of family medicine in Canada and abroad who attended the conferences. We performed a scoping review to determine the methodologic obstacles to understanding the positive effects of family medicine globally. The main obstacle to evaluating family medicine globally is that one of its core dimensions and assets is its local adaptability. Family medicine takes on very different roles in different health systems, making aggregation of data difficult. In many countries family medicine competes with other disciplines rather than performing a gatekeeping role. Further, most research that has been conducted thus far comes from industrialized contexts, and patient continuity and its benefits might not be achievable in the short term in developing countries when clinical demands are great. We must find frameworks to permit strengthening the evidentiary basis of the discipline across different contexts without sacrificing its beneficial adaptability. We believe that developing family medicine and its attributes is one of the keys to achieving global health. These attributes—including its comprehensiveness, adaptability, and attention to both local and patient needs—are key to advancing global health priorities, but make common evaluative frameworks for the discipline a challenge. The spread of family medicine over the past decades is indirect evidence of its utility, but we need to generate more evidence. We present some of the initial challenges to a broader and more rigorous evaluative framework.


Health researchers in Alberta: an exploratory comparison of defining characteristics and knowledge translation activities.
Canadian funding agencies are no longer content to support research that solely advances scientific knowledge, and key directives are now in place to promote research transfer to policy- and decision-makers. Therefore, it is necessary to improve our understanding of how researchers are trained and supported to facilitate knowledge translation activities. In this study, we investigated differences in health researcher characteristics and knowledge translation activities. Our sample consisted of 240 health researchers from three Alberta universities. Respondents were classified by research domain [basic (n = 72) or applied (n = 168)] and faculty [medical school (n = 128) or other health science (n = 112)]. We examined our findings using Mode I and Mode II archetypes of knowledge production, which allowed us to consider the scholarly and social contexts of knowledge production and translation. Differences among health researcher professional characteristics were not statistically significant. There was a significant gender difference in the applied researcher faculty group, which was predominantly female (p < .05). Research domain was linked to translation activities. Applied researchers reported engaging in significantly more Mode II activities than basic researchers (p < .001), and scored higher than basic researchers regarding the perceived importance of translation activities (Mode I, p = .01; Mode II, p < .001). Main effects of faculty were limited to engaged dissemination (medical school < other faculties; p = .025) and number of publications (medical school > other faculties; p = .004). There was an interaction effect for research domain and faculty group for number of publications (p = .01), in that applied researchers in medical faculties published more than their peers in other faculty groups. Our findings illustrate important differences between health researchers and provide beginning insights into their professional characteristics and engagement in Mode I and Mode II activities. A future study designed to examine these dimensions in greater detail, including potential covariates across more varied institutions, would yield richer insights and enable an examination of relative influences, needs and costs of each mode of activity.


Diabetes mellitus and risk of age-related macular degeneration: a systematic review and meta-analysis.
Age-related macular degeneration (AMD) is a major cause of severe vision loss in elderly people. Diabetes mellitus is a common endocrine disorder with serious consequences, and diabetic retinopathy (DR) is the main ophthalmic complication. DR and AMD are different diseases and we seek to explore the relationship between diabetes and AMD. MEDLINE, EMBASE, and the Cochrane Library were searched for potentially eligible studies. Studies based on longitudinal cohort, cross-sectional, and case-control associations, reporting evaluation data of diabetes as an independent factor for AMD were included. Reports of relative risks (RRs), hazard ratios (HRs), odds ratio (ORs), or evaluation data of diabetes as an independent factor for AMD were included. Review Manager and STATA were used for the meta-analysis. Twenty four articles involving 27 study populations were included for meta-analysis. In 7 cohort studies, diabetes was shown to be a risk factor for AMD (OR, 1.05; 95% CI, 1.00-1.14). Results of 9 cross-sectional studies revealed consistent association of diabetes with AMD (OR, 1.21; 95% CI, 1.00-1.45), especially for late AMD (OR, 1.48; 95% CI, 1.44-1.51). Similar association was also detected for AMD (OR, 1.29; 95% CI, 1.13-1.49) and late AMD (OR, 1.16; 95% CI, 1.11-1.21) in 11 case-control studies. The pooled ORs for risk of neovascular AMD (nAMD) were 1.10 (95% CI, 0.96-1.26), 1.48 (95% CI, 1.44-1.51), and 1.15 (95% CI, 1.11-1.21) from cohort, cross-sectional and case-control studies, respectively. No obvious divergence existed among different ethnic groups. Therefore, we find diabetes a risk factor for AMD, stronger for late AMD than earlier stages. However, most of the included studies only adjusted for age and sex; we thus cannot rule out confounding as a potential explanation for the association. More well-designed prospective cohort studies are still warranted to further examine the association.


Genetic markers of bone and joint health and physical capability in older adults: the HALCyon programme.
Good bone and joint health is essential for the physical tasks of daily living and poorer indicators of physical capability in older adults have been associated with increased mortality rates. Genetic variants of indicators of bone and joint health may be associated with measures of physical capability. As part of the Healthy Ageing across the Life Course (HALCyon) programme, men and women aged between 52 and 90+ years from six UK cohorts were genotyped for a polymorphism associated with serum calcium (rs1801725, CASR), two polymorphisms associated with bone mineral density (BMD) (rs2941740, ESR1 and rs9594759, RANKL) and one associated with osteoarthritis risk rs3815148 (COG5). Meta-analysis was used to pool within-study effects of the associations between each of the polymorphisms and measures of physical capability: grip strength, timed walk or get up and go, chair rises and standing balance. Few important associations were observed among the several tests. We found that carriers of the serum calcium-raising allele had poorer grip strength compared with non-carriers (pooled p=0.05, n=11,239) after adjusting for age and sex. Inconsistent results were observed for the two variants associated with BMD and we found no evidence for an association between rs3815148 (COG5) and any of the physical capability measures. Our findings suggest elevated serum calcium levels may lead to lower grip strength, though this requires further replication. Our results do not provide evidence for a substantial influence of these variants in ESR1, RANKL and COG5 on physical capability in older adults.


Independent spectral representations of images for recognition.
In recent years, studies have shown that independent components of local windows of natural images resemble the receptive fields of cells in the early stages of the mammalian visual pathway. However, the role of the independence in visual recognition is not well understood. We argue that the independence resolves the curse of dimensionality by reducing the complexity of probability models to the linear order of the dimension. In addition, we show empirically that the complexity reduction does not degrade the recognition performance on all the data sets that we have used with an independent spectral representation. In this representation, an input image is first decomposed into independent channels given by the estimated independent components from training images, and each channel's response is then summarised by using its histogram as an estimate of the underlying probability model along that dimension. We demonstrate the sufficiency of the proposed representation for image characterization by synthesizing textures and objects through sampling and for recognition by applying it to large data sets. Our comparisons show that the independent spectral representation often gives improved recognition performance.


Aging with HIV: a practical review.
The worldwide elderly population is expected to grow by an additional 694 million people by 2025. By that time, there will be approximately two billion elderly people in the world, most of whom (80%) will be living in developing countries. Based on recent estimates, this population will number over 40 million in 2030 in Brazil and a consequent increase in governmental spending for this population can be expected. Since highly active antiretroviral therapy became available in the mid-1990s, the life expectancy of people living with HIV has increased significantly. Approximately 12 million life years were added to the world between 1996 and 2008 as a consequence of wider access to highly active antiretroviral therapy. In Brazil, the incidence of AIDS among the population aged ≥50 years doubled between 1996 and 2006. The development of antiretroviral therapy has allowed individuals diagnosed at a younger age to live longer, which partially explains the aging tendency associated with the HIV/AIDS epidemic. It is estimated that by 2015, subjects aged ≥50 years will represent 50% of the people living with HIV undergoing clinical treatment. This scenario presents some challenges, including the fact that the diagnosis of HIV tends to be delayed in older patients compared to younger patients because the symptoms of HIV can be confused with those of other common diseases among the elderly and also because healthcare professionals do not consider this population to be at high risk for HIV infection. In regard to the individuals diagnosed with HIV, a further challenge is presented by the morbidity normally associated with aging. Finally, the elderly also exhibit higher susceptibility to the toxic effects and pharmacological interactions of medications. The present article reviews the literature regarding the profile of HIV infection among individuals aged ≥50 years focusing on practical features related to the clinical approach and long-term follow-up of this population.


Special topics--Mitigation of methane and nitrous oxide emissions from animal operations: III. A review of animal management mitigation options.
The goal of this review was to analyze published data on animal management practices that mitigate enteric methane (CH4) and nitrous oxide (N2O) emissions from animal operations. Increasing animal productivity can be a very effective strategy for reducing greenhouse gas (GHG) emissions per unit of livestock product. Improving the genetic potential of animals through planned cross-breeding or selection within breeds and achieving this genetic potential through proper nutrition and improvements in reproductive efficiency, animal health, and reproductive lifespan are effective approaches for improving animal productivity and reducing GHG emission intensity. In subsistence production systems, reduction of herd size would increase feed availability and productivity of individual animals and the total herd, thus lowering CH4 emission intensity. In these systems, improving the nutritive value of low-quality feeds for ruminant diets can have a considerable benefit on herd productivity while keeping the herd CH4 output constant or even decreasing it. Residual feed intake may be a tool for screening animals that are low CH4 emitters, but there is currently insufficient evidence that low residual feed intake animals have a lower CH4 yield per unit of feed intake or animal product. Reducing age at slaughter of finished cattle and the number of days that animals are on feed in the feedlot can significantly reduce GHG emissions in beef and other meat animal production systems. Improved animal health and reduced mortality and morbidity are expected to increase herd productivity and reduce GHG emission intensity in all livestock production systems. Pursuing a suite of intensive and extensive reproductive management technologies provides a significant opportunity to reduce GHG emissions. Recommended approaches will differ by region and species but should target increasing conception rates in dairy, beef, and buffalo, increasing fecundity in swine and small ruminants, and reducing embryo wastage in all species. Interactions among individual components of livestock production systems are complex but must be considered when recommending GHG mitigation practices.


Nongenomic mechanisms of physiological estrogen-mediated dopamine efflux.
Neurological diseases and neuropsychiatric disorders that vary depending on female life stages suggest that sex hormones may influence the function of neurotransmitter regulatory machinery such as the dopamine transporter (DAT). In this study we tested the rapid nongenomic effects of several physiological estrogens [estradiol (E2), estrone (E1), and estriol (E3)] on dopamine efflux via the DAT in a non-transfected, NGF-differentiated, rat pheochromocytoma (PC12) cell model that expresses membrane estrogen receptors (ERs) alpha, beta, and GPR30. We examined kinase, ionic, and physical interaction mechanisms involved in estrogenic regulation of the DAT function. E2-mediated dopamine efflux is DAT-specific and not dependent on extracellular Ca2+-mediated exocytotic release from vesicular monoamine transporter vesicles (VMATs). Using kinase inhibitors we also showed that E2-mediated dopamine efflux is dependent on protein kinase C and MEK activation, but not on PI3K or protein kinase A. In plasma membrane there are ligand-independent associations of ERalpha and ERbeta (but not GPR30) with DAT. Conditions which cause efflux (a 9 min 10(-9) M E2 treatment) cause trafficking of ERalpha (stimulatory) to the plasma membrane and trafficking of ERbeta (inhibitory) away from the plasma membrane. In contrast, E1 and E3 can inhibit efflux with a nonmonotonic dose pattern, and cause DAT to leave the plasma membrane. Such mechanisms explain how gender biases in some DAT-dependent diseases can occur.


Simulation insights into the role of antiparallel molecular association in the formation of smectic A phases.
A simple dissipative particle dynamics (DPD) model is introduced, which can be used to represent a broad range of calamitic mesogens. The model allows for antiparallel association that occurs naturally in a number of mesogens with terminal dipoles, including the 4-n-alkyl-4'-cyanobiphenyl (nCB) series. Favourable antiparallel interactions lead to the formation of SmAd phases in which the layer spacing is intermediate between monolayer and bilayer. The model is easily tuned to vary the strength of antiparallel association and the SmA layer spacing, and to give either isotropic-smectic or isotropic-nematic-smectic phase sequences. The model allows for a range of other smectics: including SmA1 phases exhibiting microphase separation within layers, and smectics A structures with more complicated repeat units. For large system sizes (≥50 000 molecules) in the nematic phase, we are able to demonstrate the formation of three distinct types of cybotactic domains depending on the local interactions. Cybotactic domains are found to grow in the nematic-smectic pretransitional region as the system moves closer to TSN.


Bilingualism delays the onset of behavioral but not aphasic forms of frontotemporal dementia.
Bilingualism has been found to delay onset of dementia and this has been attributed to an advantage in executive control in bilinguals. However, the relationship between bilingualism and cognition is complex, with costs as well as benefits to language functions. To further explore the cognitive consequences of bilingualism, the study used Frontotemporal dementia (FTD) syndromes, to examine whether bilingualism modifies the age at onset of behavioral and language variants of Frontotemporal dementia (FTD) differently. Case records of 193 patients presenting with FTD (121 of them bilingual) were examined and the age at onset of the first symptoms were compared between monolinguals and bilinguals. A significant effect of bilingualism delaying the age at onset of dementia was found in behavioral variant FTD (5.7 years) but not in progressive nonfluent aphasia (0.7 years), semantic dementia (0.5 years), corticobasal syndrome (0.4 years), progressive supranuclear palsy (4.3 years) and FTD-motor neuron disease (3 years). On dividing all patients predominantly behavioral and predominantly aphasic groups, age at onset in the bilingual behavioral group (62.6) was over 6 years higher than in the monolingual patients (56.5, p=0.006), while there was no difference in the aphasic FTD group (60.9 vs. 60.6 years, p=0.851). The bilingual effect on age of bvFTD onset was shown independently of other potential confounding factors such as education, gender, occupation, and urban vs rural dwelling of subjects. To conclude, bilingualism delays the age at onset in the behavioral but not in the aphasic variants of FTD. The results are in line with similar findings based on research in stroke and with the current views of the interaction between bilingualism and cognition, pointing to advantages in executive functions and disadvantages in lexical tasks.


Measurement and control of pH in the aqueous interior of reverse micelles.
The encapsulation of proteins and nucleic acids within the nanoscale water core of reverse micelles has been used for over 3 decades as a vehicle for a wide range of investigations including enzymology, the physical chemistry of confined spaces, protein and nucleic acid structural biology, and drug development and delivery. Unfortunately, the static and dynamical aspects of the distribution of water in solutions of reverse micelles complicate the measurement and interpretation of fundamental parameters such as pH. This is a severe disadvantage in the context of (bio)chemical reactions and protein structure and function, which are generally highly sensitive to pH. There is a need to more fully characterize and control the effective pH of the reverse micelle water core. The buffering effect of titratable head groups of the reverse micelle surfactants is found to often be the dominant variable defining the pH of the water core. Methods for measuring the pH of the reverse micelle aqueous interior using one-dimensional (1)H and two-dimensional heteronuclear NMR spectroscopy are described. Strategies for setting the effective pH of the reverse micelle water core are demonstrated. The exquisite sensitivity of encapsulated proteins to the surfactant, water content, and pH of the reverse micelle is also addressed. These results highlight the importance of assessing the structural fidelity of the encapsulated protein using multidimensional NMR before embarking upon a detailed structural and biophysical characterization.


Successful control of methicillin-resistant Staphylococcus aureus outbreak at a university department of dermatology.
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading strains among multiresistant hospital microflora. In 2003, we noted an increase in the number of MRSA strain 35 among patients at the University of Zurich, Department of Dermatology. At the end of 2003, we implemented additional policies in hospital hygiene, which significantly decreased the number of MRSA infections. This is a retrospective study on 65 consecutive patients at the Department of Dermatology, University Hospital of Zurich, in whom MRSA contamination was newly diagnosed during the period of 2003-2008. All isolated strains were genotyped. We implemented additional policies as strict hand hygiene and avoidance of sharing the same ointment pots or tubes amongst patients. As soon as the skin disease was healed, decolonization of MRSA was undertaken by a 5-day topical treatment. Of the 65 MRSA patients, 19 (29%) patients carried a genotypical strain (MRSA 35) that was identified to be specific for the Department of Dermatology. Three health care workers (HCWs) were tested positive. The outlined measures reduced the incidence of new transmissions of this specific strain MRSA 35 significantly (P=0.001) with a complete disappearance of new transmissions of MRSA 35 in the year 2008. Of the 65 patients, 15 (23%) patients became long-term carriers, among all (15/15; 100%) had persisting active skin lesions. Strict hand hygiene and avoidance of sharing ointments among patients were highly effective measures in controlling an outbreak of MRSA. Complete of near-to-complete remission of the underlying dermatoses, skin lesions or chronic wounds, is a prerequisite for complete decolonization of MRSA carriers.


A novel pretargeting method for measuring antibody internalization in tumor cells.
A novel pretargeting method has been developed to quantitate antibody cellular internalization. In this study, the antibody was conjugated with a phosphorodiamidate morpholino oligomer (MORF) specific for the complementary MORF (cMORF) as an effector. Half the tumor cells were incubated with the MORF-antibody (pretargeting group) and the other half with the same MORF-antibody at the same concentration but radiolabeled (direct targeting group). After incubation, the same dosage of radiolabeled cMORF was added to the wells of the pretargeting group. The radioactivity of the direct targeting cells represented the sum of both internalized and cell-surface-bound antibodies, whereas the radioactivity of the pretargeting cells resulted only from the surface-bound antibodies, as the radiolabeled cMORF does not penetrate the cell surface. Therefore, the difference in radioactivity accumulation between pretargeting and direct targeting provides the internalized fraction. In this example, the internalization of a MORF conjugated anti-prostate-specific membrane antigen antibody, 3C6, in LNCaP cells was examined, and the average cell-surface residence time was determined as 2 hours. This method of measuring antibody internalization is directly applicable to pretargeting applications but can be a universal alternative to the conventional acid-wash method, with the advantage of leaving the cell membrane undamaged.


Delayed resolution of lung inflammation in Il-1rn-/- mice reflects elevated IL-17A/granulocyte colony-stimulating factor expression.
IL-1 has been associated with acute lung injury (ALI) in both humans and animal models, but further investigation of the precise mechanisms involved is needed, and may identify novel therapeutic targets. To discover the IL-1 mediators essential to the initiation and resolution phases of acute lung inflammation, knockout mice (with targeted deletions for either the IL-1 receptor-1, i.e., Il-1r1(-/-), or the IL-1 receptor antagonist, i.e., Il-1rn(-/-)) were exposed to aerosolized LPS, and indices of lung and systemic inflammation were examined over the subsequent 48 hours. The resultant cell counts, histology, protein, and RNA expression of key cytokines were measured. Il-1r1(-/-) mice exhibited decreased neutrophil influx, particularly at 4 and 48 hours after exposure to LPS, as well as reduced bronchoalveolar lavage (BAL) expression of chemokines and granulocyte colony-stimulating factor (G-CSF). On the contrary, Il-1rn(-/-) mice demonstrated increased BAL neutrophil counts, increased BAL total protein, and greater evidence of histologic injury, all most notably 2 days after LPS exposure. Il-1rn(-/-) mice also exhibited higher peripheral neutrophil counts and greater numbers of granulocyte receptor-1 cells in their bone marrow, potentially reflecting their elevated plasma G-CSF concentrations. Furthermore, IL-17A expression was increased in the BAL and lungs of Il-1rn(-/-) mice after exposure to LPS, likely because of increased numbers of γδ T cells in the Il-1rn(-/-) lungs. Blockade with IL-17A monoclonal antibody before LPS exposure decreased the resultant BAL neutrophil counts and lung G-CSF expression in Il-1rn(-/-) mice, 48 hours after exposure to LPS. In conclusion, Il-1rn(-/-) mice exhibit delayed resolution in acute lung inflammation after exposure to LPS, a process that appears to be mediated via the G-CSF/IL-17A axis.


The plasticity of the mirror system: how reward learning modulates cortical motor simulation of others.
Cortical motor simulation supports the understanding of others' actions and intentions. This mechanism is thought to rely on the mirror neuron system (MNS), a brain network that is active both during action execution and observation. Indirect evidence suggests that (alpha/beta) mu suppression, an electroencephalographic (EEG) index of MNS activity, is modulated by reward. In this study we aimed to test the plasticity of the MNS by directly investigating the link between (alpha/beta) mu suppression and reward. 40 individuals from a general population sample took part in an evaluative conditioning experiment, where different neutral faces were associated with high or low reward values. In the test phase, EEG was recorded while participants viewed videoclips of happy expressions made by the conditioned faces. Alpha/beta mu suppression (identified using event-related desynchronisation of specific independent components) in response to rewarding faces was found to be greater than for non-rewarding faces. This result provides a mechanistic insight into the plasticity of the MNS and, more generally, into the role of reward in modulating physiological responses linked to empathy.


Differential regulation of the expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase, synthase, and low density lipoprotein receptor genes.
The ability of mitogenic stimulation of human T lymphocytes to alter the expression of genes involved in sterol metabolism was examined. Messenger RNA levels for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, HMG-CoA synthase, and low density lipoprotein (LDL) receptor were quantified in resting and mitogen-stimulated T lymphocytes by nuclease protection assay. Mitogenic stimulation increased HMG-CoA synthase mRNA levels by 5-fold and LDL receptor by 4-fold when cells were cultured in lipoprotein-depleted medium whereas HMG-CoA reductase gene expression was not significantly increased. When cultures were supplemented with concentrations of low density lipoprotein sufficient to saturate LDL receptors, expression of all three genes was inhibited in resting lymphocytes, as effectively as was noted with fibroblasts. Similarly, LDL down-regulated gene expression in mitogen-activated lymphocytes so that mitogenic stimulation did not increase either HMG-CoA reductase or synthase mRNA levels, although LDL receptor gene expression was enhanced. These results indicate that expression of three of the genes involved in sterol metabolism is differentially regulated by LDL and mitogenic stimulation. Moreover, the increase in rates of endogenous sterol synthesis and the activity of HMG-CoA reductase in mitogen-stimulated T lymphocytes cannot be accounted for by increases in HMG-CoA reductase mRNA levels.


Lys656Asn polymorphism of leptin receptor gene and metabolic syndrome in obese patients.
The etiology of common obesity is complex, because many genetic, environmental and metabolic factors might act. Alterations of the normal leptin receptor gene be involved in the development of obesity. The polymorphism on codon 656 produces a change in charge, making this change a possibility to be functional. The aim of our study was to investigate the relationship between metabolic syndrome and Lys656Asn polymorphism in obese patients. A population of 714 obese patients (body mass index > 30) was analyzed in cross-sectional survey. A bioimpedance, blood pressure, a serial assessment of nutritional intake with 3 days written food records and biochemical analysis were performed. Four hundred and seventy eight patients (66.9%) had the genotype Lys656/Lys 656 (wild group), whereas 236 (33.1%) had either the genotype Lys656/Asn656 (212 patients, 29.7%) or the genotype Asn656/Asn656 (24 patients, 3.4%) (mutant group). Prevalence of metabolic syndrome (MS) with ATP III definition was 49.4% (353 patients; 35.1% males and 64.9% females) and 50.6% patients without MS (n = 361; 25.2% males and 75.8% females). Prevalence of leptin receptor (LEPR) genotypes was similar in patients with metabolic syndrome (65.5% wild genotype and 34.5% mutant genotype) and without metabolic syndrome (68.3% wild genotype and 31.7% mutant genotype). No differences in anthropometric and biochemical parameters were detected between genotypes in the same group of metabolic syndrome. The finding of our study is the lack of association of the Lys656/Asn656 and Asn656/ Asn656 genotypes with metabolic syndrome.


Screening for psychological difficulties in young children in crisis: complementary cross-cultural validation.
Detection of children's psychological difficulties in crises and low resource settings is challenged by the lack of a validated, rapid and simple tool. We present the results of two confirmatory validations of the Psychological Screening for Young Children aged 3 to 6 years (PSYCa 3-6) scale. We performed cross-cultural validations, assessing the performance of the scale in different contexts. These were conducted in Mathare, Nairobi, Kenya and Buenaventura, Colombia between December 2009 and February 2012. External validity was assessed comparing the PSYCa 3-6 against a clinical interview and the Clinical Global Impression Severity scale (CGI). A total of 160 mothers or caregivers of children 3 to 6 years old in Mathare and 148 in Buenaventura were included in the study. Both demonstrated good concurrent validity (Buenaventura ρ=0.49, p<0.0001; Mathare ρ=0.41, p<0.0001). Inter-rater reliability was found to be acceptable in Buenaventura (intraclass correlation [ICC]=0.69 [0.4-0.84]) and high in Mathare (0.87 [0.75-0.94]). As shown by its validation in diverse contexts, use in other populations may help improve the delivery of mental health care to children in crises and low-resource settings. Additional research on the design and delivery of intervention models for crises remains essential.


A statement synthesis of emotional eating and body size recognition: advancing nursing science related to obesity research.
A discussion of a statement synthesis of the relationship between the concepts of emotional eating and body size recognition. The interrelatedness of the concepts of emotional eating and body size recognition is poorly understood; however, both factors significantly impact weight management. Nurses can be instrumental in developing more effective weight management interventions as a critical step in helping to ameliorate the public health burden of obesity. This discussion article reports a literary statement synthesis. Five quantitative research studies from 1996 - 2014 were identified, which examined the relationship between the concepts. A relational statement delineating the direct association between the amount of self-reported emotional eating and an adult individual's recognition of their body size was constructed. Construction of a relational statement about emotional eating and recognition of body size is a step in the nursing theory-building process. The constructed relational statement will guide future research and nursing theory development and may ultimately help to construct more effective weight management interventions.


Carpooling and booster seats: a national survey of parents.
Booster seat use among school-aged children has been consistently lower than national goals. In this study, we sought to explore associations between parental experiences with booster seats and carpooling. We conducted a cross-sectional Web-based survey of a nationally representative panel of US parents in January 2010. As part of a larger survey, parents of 4- to 8-year-old children responded to 12 questions related to booster seats and carpooling. Of 1612 parents responding to the full survey (response rate = 71%), 706 had a 4- to 8-year-old child and 681 met inclusion rules. Most parents (76%) reported their child used a safety seat when riding in the family car. Of children reported to use seat belts, 74% did so in accordance with their state law. Parent report of child safety seat use was associated with younger child age and with the presence of state booster seat laws. Sixty-four percent of parents carpool. Among parents who carpool and whose children use a child safety seat: 79% indicated they would always ask another driver to use a booster seat for their child and 55% reported they always have their child use their booster seat when driving friends who do not have boosters. Carpooling is a common driving situation during which booster seat use is inconsistent. Social norms and self-efficacy are associated with booster seat use. Clinicians who care for children should increase efforts to convey the importance of using the size-appropriate restraint for every child on every trip.


Didymocarpus lobulatus (Gesneriaceae), a new species from Zhejiang Province, East China.
Didymocarpus lobulatus, a new species endemic to Zhejiang province, eastern China, is described and illustrated with photographs. The new species is morphologically similar to D. heucherifolius, D. cortusifolius and D. salviiflorus in leaf morphology, but can be easily distinguished by a combination of characters, including the shape of bracts, calyx and calyx lobes.


The emerging role of the phosphatidylinositol 3-kinase/ akt/mammalian target of rapamycin signaling network in cancer stem cell biology.
The cancer stem cell theory entails the existence of a hierarchically organized, rare population of cells which are responsible for tumor initiation, self-renewal/maintenance, and mutation accumulation. The cancer stem cell proposition could explain the high frequency of cancer relapse and resistance to currently available therapies. The phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway regulates a wide array of physiological cell functions which include differentiation, proliferation, survival, metabolism, autophagy, and motility. Dysregulated PI3K/Akt/mTOR signaling has been documented in many types of neoplasias. It is now emerging that this signaling network plays a key role in cancer stem cell biology. Interestingly, cancer stem cells displayed preferential sensitivity to pathway inhibition when compared to healthy stem cells. This observation provides the proof-of-principle that functional differences in signaling pathways between neoplastic stem cells and healthy stem cells could be identified. In this review, we present the evidence which links the signals emanating from the PI3K/Akt/mTOR cascade with the functions of cancer stem cells, both in solid and hematological tumors. We then highlight how targeting PI3K/Akt/mTOR signaling with small molecules could improve cancer patient outcome.


Modulation of renal type IIa Na+/Pi cotransporter kinetics by the arginine modifier phenylglyoxal.
The effects of the arginine-modifying reagent phenylglyoxal on the kinetics of the type IIa Na + /Pi cotransporter expressed in Xenopus, oocytes were studied by means of 32Pi uptake and electrophysiology. Phenylglyoxal incubation induced up to 60% loss of cotransport function but only marginally altered the Na+-leak. Substrate activation and pH dependency remained essentially unaltered, whereas the voltage dependency of Pi-induced change in electrogenic response was significantly reduced. Presteady-state charge movements were suppressed and the equilibrium charge distribution was shifted slightly towards hyperpolarizing potentials. Charge movements in the absence of external Na+ were also suppressed, which indicated that the empty-carrier kinetics were modified. These effects were incorporated into an ordered alternating access model for NaPi-IIa, whereby the arginine modification by phenylglyoxal was modeled as altered apparent electrical distances moved by mobile charges, together with a slower rate of translocation of the electroneutral, fully loaded carrier.


Suicidal Behavior in Juvenile Delinquents: The Role of ADHD and Other Comorbid Psychiatric Disorders.
This study evaluated the role of psychiatric morbidity in relation to a history of suicidal behavior, with a particular focus on attention-deficit/hyperactivity disorder (ADHD). Suicidality and psychiatric diagnoses were assessed in 370 incarcerated male juvenile delinquents from Northern Russia using the semi-structured K-SADS-PL psychiatric interview. A lifetime history of suicidal ideation only (24.7 %) and suicidal ideation with suicide attempts (15.7 %) was common. Binary logistic regression analysis was used to assess the role of ADHD and other psychiatric disorders in suicidal ideation and suicide attempts. A history of suicidal ideation and of suicide attempts were associated with higher rates of psychiatric morbidity and with the number of comorbid psychiatric disorders. An ADHD diagnosis was associated with an increased risk for both suicidal ideation and for suicide attempts. The comorbidity of ADHD with drug dependence further increased the risk for suicidal ideation, while ADHD and alcohol dependence comorbidity increased the risk for suicide attempts. Our findings highlight the importance of adequately detecting and treating psychiatric disorders in vulnerable youths, especially when they are comorbid with ADHD.


Influence of ESAT-6 secretion system 1 (RD1) of Mycobacterium tuberculosis on the interaction between mycobacteria and the host immune system.
The chromosomal locus encoding the early secreted antigenic target, 6 kDa (ESAT-6) secretion system 1 of Mycobacterium tuberculosis, also referred to as "region of difference 1 (RD1)," is absent from Mycobacterium bovis bacillus Calmette-Guerin (BCG). In this study, using low-dose aerosol infection in mice, we demonstrate that BCG complemented with RD1 (BCG::RD1) displays markedly increased virulence which albeit does not attain that of M. tuberculosis H37Rv. Nevertheless, phenotypic and functional analyses of immune cells at the site of infection show that the capacity of BCG::RD1 to initiate recruitment/activation of immune cells is comparable to that of fully virulent H37Rv. Indeed, in contrast to the parental BCG, BCG::RD1 mimics H37Rv and induces substantial influx of activated (CD44highCD45RB(-)CD62L(-)) or effector (CD45RB(-)CD27(-)) T cells and of activated CD11c(+)CD11bhigh cells to the lungs of aerosol-infected mice. For the first time, using in vivo analysis of transcriptome of inflammatory cytokines and chemokines of lung interstitial CD11c+ cells, we show that in a low-dose aerosol infection model, BCG::RD1 triggered an activation/inflammation program comparable to that induced by H37Rv while parental BCG, due to its overattenuation, did not initiate the activation program in lung interstitial CD11c+ cells. Thus, products encoded by the ESAT-6 secretion system 1 of M. tuberculosis profoundly modify the interaction between mycobacteria and the host innate and adaptive immune system. These modifications can explain the previously described improved protective capacity of BCG::RD1 vaccine candidate against M. tuberculosis challenge.


A mass spectrometric and quantum chemical study of the vaporisation of lead monoxide in a flow of gaseous arsenic and antimony trioxides.
Mass spectrometric studies of the vapours over solid lead oxide in a flow of gaseous arsenic and antimony trioxides were conducted. The following ions of the ternary oxides were detected: Pb3As2O6(+), Pb3AsO4(+), PbAs2O4(+), PbAsO2(+), PbSb2O4(+), and PbSbO2(+). The origin of these species produced by the ionisation and/or fragmentation of ternary gaseous oxides is discussed. The PbAs2O4 species was undoubtedly identified by the determination of the appearance energy. Presumably, the Pb3As2O6 and PbSb2O4 species also existed in the gas phase. Thermodynamic data for the ternary oxides were obtained experimentally by means of a mass spectrometric Knudsen-cell method and were confirmed by quantum chemical calculations.


Confocal microscopy and multi-photon excitation microscopy of human skin in vivo.
This paper compares tandem scanning reflected light confocal microscopy and multi-photon excitation microscopy for the observation of human skin in vivo. Tandem scanning confocal light microscopy based on a white light source can provide video-rate image acquisition from the skin surface to the epidermal-dermal junction. Multi-photon excitation is induced by a 80 MHz pulse train of femtosecond laser pulses at 780 nm wavelength. This nonlinear microscopic technique is inherently suitable for deep tissue fluorescence imaging. The relative merits of these two techniques can be identified by comparing movies of optical sections obtained from the forearm skin of the same volunteer.


An eye movement corpus study of the age-of-acquisition effect.
In the present study, we investigated the effects of word-level age of acquisition (AoA) on natural reading. Previous studies, using multiple language modalities, showed that earlier-learned words are recognized, read, spoken, and responded to faster than words learned later in life. Until now, in visual word recognition the experimental materials were limited to single-word or sentence studies. We analyzed the data of the Ghent Eye-tracking Corpus (GECO; Cop, Dirix, Drieghe, & Duyck, in press), an eyetracking corpus of participants reading an entire novel, resulting in the first eye movement megastudy of AoA effects in natural reading. We found that the ages at which specific words were learned indeed influenced reading times, above other important (correlated) lexical variables, such as word frequency and length. Shorter fixations for earlier-learned words were consistently found throughout the reading process, in both early (single-fixation durations, first-fixation durations, gaze durations) and late (total reading times) measures. Implications for theoretical accounts of AoA effects and eye movements are discussed.


Quantitative Impact of Cell Membrane Fluorescence Labeling on Phagocytosis Measurements in Confrontation Assays.
Phagocytosis is series of steps where the pathogens and the immune cells interact during an invasion. This starts with the adhesion process between the host and pathogen cells, and is followed by the engulfment of the pathogens. Many analytical methods that are applied to characterize phagocytosis based on imaging the host-pathogen confrontation assays rely on the fluorescence labeling of cells. However, the potential effect of the membrane labeling on the quantitative results of the confrontation assays has not been studied in detail. In this study, we determine whether the fluorescence labeling processes themselves influence the results of the phagocytosis measurements. Here, alveolar macrophages, which form one of the most important compartments of the innate immune system, were used as an example of host cells, whereas Aspergillus fumigatus and Lichtheimia corymbifera that cause aspergillosis and mucormycosis, respectively, were studied as examples for pathogens. At first, our study investigated the importance of the sequence of steps of the fixation process when preparing the confrontation assay sample for microscopy studies. Here we showed that applying the fixation agent before the counter-staining causes miscalculations during the determination of the phagocytic measures. Furthermore, we also found that staining the macrophages with various concentrations of DID, as a typical membrane label, in most cases altered the capability of macrophages to phagocytose FITC-stained A. fumigatus and L. corymbifera spores in comparison with unlabeled macrophages. This effect of the DID staining showed a differential character dependent upon the labeling status and the specific type of pathogen. Moreover, labeling the spores of A. fumigatus and L. corymbifera with FITC increased the phagocytic measures during confrontation with unlabeled macrophages when compared to label-free spores. Overall, our study confirms that the staining process itself may significantly manipulate the quantitative outcome of the confrontation assay. As a result of our study, we also developed a user-friendly image analysis tool that analyses confrontation assays both with and without fluorescence labeling of the host cells and of the pathogens. Our image analysis algorithm saves experimental work effort and time, provides more precise results when calculating the phagocytic measures, and delivers a convenient analysis tool for the biologists to monitor host-pathogen interactions as they happen without the artifacts that fluorescence labeling imposes on biological interactions.


The anti-sigma factor MucA of Pseudomonas aeruginosa: Dramatic differences of a mucA22 vs. a ΔmucA mutant in anaerobic acidified nitrite sensitivity of planktonic and biofilm bacteria in vitro and during chronic murine lung infection.
Mucoid mucA22 Pseudomonas aeruginosa (PA) is an opportunistic lung pathogen of cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) patients that is highly sensitive to acidified nitrite (A-NO2-). In this study, we first screened PA mutant strains for sensitivity or resistance to 20 mM A-NO2- under anaerobic conditions that represent the chronic stages of the aforementioned diseases. Mutants found to be sensitive to A-NO2- included PA0964 (pmpR, PQS biosynthesis), PA4455 (probable ABC transporter permease), katA (major catalase, KatA) and rhlR (quorum sensing regulator). In contrast, mutants lacking PA0450 (a putative phosphate transporter) and PA1505 (moaA2) were A-NO2- resistant. However, we were puzzled when we discovered that mucA22 mutant bacteria, a frequently isolated mucA allele in CF and to a lesser extent COPD, were more sensitive to A-NO2- than a truncated ΔmucA deletion (Δ157-194) mutant in planktonic and biofilm culture, as well as during a chronic murine lung infection. Subsequent transcriptional profiling of anaerobic, A-NO2--treated bacteria revealed restoration of near wild-type transcript levels of protective NO2- and nitric oxide (NO) reductase (nirS and norCB, respectively) in the ΔmucA mutant in contrast to extremely low levels in the A-NO2--sensitive mucA22 mutant. Proteins that were S-nitrosylated by NO derived from A-NO2- reduction in the sensitive mucA22 strain were those involved in anaerobic respiration (NirQ, NirS), pyruvate fermentation (UspK), global gene regulation (Vfr), the TCA cycle (succinate dehydrogenase, SdhB) and several double mutants were even more sensitive to A-NO2-. Bioinformatic-based data point to future studies designed to elucidate potential cellular binding partners for MucA and MucA22. Given that A-NO2- is a potentially viable treatment strategy to combat PA and other infections, this study offers novel developments as to how clinicians might better treat problematic PA infections in COPD and CF airway diseases.


Professionalism Redundant, Reshaped, or Reinvigorated? Realizing the "Third Logic" in Contemporary Health Care.
Recent decades have seen the influence of the professions decline. Lately, commentators have suggested a revived role for a "new" professionalism in ensuring and enhancing high-quality health care in systems dominated by market and managerial logics. The form this new professionalism might take, however, remains obscure. This article uses data from an ethnographic study of three English health care improvement projects to analyze the place, potential, and limitations of professionalism as a means of engaging clinicians in efforts to improve service quality. We found that appeals to notions of professionalism had strong support among practitioners, but converting enthusiasm for the principle of professionalism into motivation to change practice was not straightforward. Some tactics used in pursuit of this deviated sharply from traditional models of collegial social control. In systems characterized by fissures between professional groups and powerful market and managerial influences, we suggest that professionalism must interact creatively but carefully with other logics.


Lessons from the severe acute respiratory syndrome outbreak in Hong Kong.
Severe acute respiratory syndrome (SARS) is now a global public health threat with many medical, ethical, social, economic, political, and legal implications. The nonspecific signs and symptoms of this disease, coupled with a relatively long incubation period and the initial absence of a reliable diagnostic test, limited the understanding of the magnitude of the outbreak. This paper outlines our experience with public health issues that have arisen during this outbreak of SARS in Hong Kong. We confirmed that case detection, reporting, clear and timely dissemination of information, and strict infection control measures are essential in handling such an infectious disease outbreak. The need for an outbreak response unit is crucial to combat any future outbreak.


The dissociation between command following and communication in disorders of consciousness: an fMRI study in healthy subjects.
Neuroimaging studies have identified a subgroup of patients with a Disorder of Consciousness (DOC) who, while being behaviorally non-responsive, are nevertheless able to follow commands by modulating their brain activity in motor imagery (MI) tasks. These techniques have even allowed for binary communication in a small number of DOC patients. However, the majority of patients who can follow commands are unable to use their responses to communicate. A similar dissociation between present command following (CF) and absent communication abilities has been reported in overt behavioral assessments. However, the neural correlates of this dissociation in both overt and covert modalities are unknown. Here, we used functional magnetic resonance imaging (fMRI) to explore the neural mechanisms underlying CF and selection of responses for binary communication using either executed or imagined movements. Fifteen healthy participants executed or imagined two different types of arm movements that were either pre-determined by the experimenters (CF) or decided by them (action selection, AS). Action selection involved greater activity in high-level associative areas in frontal and parietal regions than CF. Additionally, motor execution (ME), as compared to MI, activated contralateral motor cortex, while the opposite contrast revealed activation in the ipsilateral sensorimotor cortex and the left inferior frontal gyrus. Importantly, there was no interaction between the task (CF/AS) and modality (MI/ME). Our results suggest that the neural processes involved in following a motor command or selecting between two motor actions are not dependent on how the response is expressed (via ME/MI). They also suggest a potential neural basis for the distinction in cognitive abilities seen in DOC patients.


A single subunit, Dis3, is essentially responsible for yeast exosome core activity.
The conserved core of the exosome, the major eukaryotic 3' --> 5' exonuclease, contains nine subunits that form a ring similar to the phosphorolytic bacterial PNPase and archaeal exosome, as well as Dis3. Dis3 is homologous to bacterial RNase II, a hydrolytic enzyme. Previous studies have suggested that all subunits are active 3' --> 5' exoRNases. We show here that Dis3 is responsible for exosome core activity. The purified exosome core has a hydrolytic, processive and Mg(2+)-dependent activity with characteristics similar to those of recombinant Dis3. Moreover, a catalytically inactive Dis3 mutant has no exosome core activity in vitro and shows in vivo RNA degradation phenotypes similar to those resulting from exosome depletion. In contrast, mutations in Rrp41, the only subunit carrying a conserved phosphorolytic site, appear phenotypically not different from wild-type yeast. We observed that the yeast exosome ring mediates interactions with protein partners, providing an explanation for its essential function.


Convergent evolution in feeding types: salivary gland mass differences in wild ruminant species.
In the ongoing debate about divergent evolutionary morphophysiological adaptations of grazing and browsing ruminants, the size of the salivary glands has received special attention. Here, we report the most comprehensive dataset on ruminant salivary glands so far, with data on the Glandula parotis (n=62 species), Gl. mandibularis (n=61), Gl. buccalis ventralis (n=44), and Gl. sublingualis (n=30). All four salivary gland complexes showed allometric scaling with body mass (BM); in all cases, the 95% confidence interval for the allometric exponent included 0.75 but did not include 1.0 (linearity); therefore, like other parameters linked to the process of food intake, salivary gland mass appears to be correlated to metabolic body weight (BM0.75), and comparisons of relative salivary gland mass between species should rather be made on the basis of BM0.75 than as a percentage of BM. In the subsequent analyses, the percentage of grass (%grass) in the natural diet was used to characterize the feeding type; the phylogenetic tree used for a controlled statistical evaluation was entirely based on mitochondrial DNA information. Regardless of phylogenetic control in the statistical treatment, there was, for all four gland complexes, a significant positive correlation of BM and gland mass, and a significant negative correlation between %grass in the natural diet and gland mass. If the Gl. parotis was analyzed either for cervid or for bovid species only, the negative correlation of gland mass and %grass was still significant in either case; an inspection of certain ruminant subfamilies, however, suggested that a convergent evolutionary adaptation can only be demonstrated if a sufficient variety of ruminant subfamilies are included in a dataset. The results support the concept that ruminant species that ingest more grass have smaller salivary glands, possibly indicating a reduced requirement for the production of salivary tannin-binding proteins.


Chronic nitrogen addition induces a cascade of plant community responses with both seasonal and progressive dynamics.
Short-lived herbaceous plants provide a useful model to rapidly reveal how multiple generations of plants in natural plant communities of sensitive desert ecosystems will be affected by N deposition. We monitored dynamic responses of community structure, richness, evenness, density and biomass of herbaceous plants to experimental N addition (2:1 NH4+:NO3- added at 0, 0.5, 1, 3, 6 and 24gNm-2a-1) in three seasons in each of three years in the Gurbantunggut desert, a typical temperate desert of central Asia. We found clear rate-dependent and season-dependent effects of N deposition on each of these variables, in most cases becoming more obvious through time. N addition reduced plant richness, leading to a loss of about half of the species after three generations in the highest N application level. Evenness and density were relatively insensitive to all but the greatest levels of N addition for two generations, but negative effects emerged in the third generation. Biomass, both above and below ground, was non-linearly affected by N deposition. Low and intermediate levels of N deposition often increased biomass, whereas the highest level suppressed biomass. Stimulatory effects of intermediate N addition disappeared in the third generation. All of these responses are strongly interrelated in a cascade of changes. Notably, changes in biomass due to N deposition were mediated by declines in richness and evenness, and other changes in community structure, rather than solely being the direct outcome of release from limitation. The interrelationships between N deposition and the different plant community attributes change not only seasonally, but also progressively change through time. These temporal changes appear to be largely independent of interannual or seasonal climatic conditions.


Many faces of pulmonary aspergillosis.
Aspergillus is a ubiquitous fungus. It is commonly isolated as an upper respiratory tract saprophyte and is the most frequent contaminant in laboratory specimens. Because species of aspergillus are omnipresent, one must be cautious in ascribing a causal role to the fungus obtained from patients. Aspergillus has low pathogenicity for humans and animals and rarely invades the immunologically competent host. Although the fungus can affect any organ system, the respiratory tract is involved in >90% of afflicted patients. Pulmonary aspergillosis, depending whether the host is atopic or immunosuppressed, may be classified under four categories: allergic or hypersensitivity syndromes, saprophytic colonization, invasive (infective) dissemination, and chemical or toxic pneumonitis. These entities differ clinically, radiologically, immunologically, and in their response to various therapeutic agents. An increased awareness, an aggressive approach to securing the diagnosis, and instituting early and appropriate therapy are needed to reduce the high morbidity and mortality caused by many of the aspergillus-related syndromes.


Composition, Spatial Characteristics, and Prognostic Significance of Myeloid Cell Infiltration in Pancreatic Cancer.
Although abundant myeloid cell populations in the pancreatic ductal adenocarcinoma (PDAC) microenvironment have been postulated to suppress antitumor immunity, the composition of these populations, their spatial locations, and how they relate to patient outcomes are poorly understood. To generate spatially resolved tumor and immune cell data at single-cell resolution, we developed two quantitative multiplex immunofluorescence assays to interrogate myeloid cells (CD15, CD14, ARG1, CD33, HLA-DR) and macrophages [CD68, CD163, CD86, IFN regulatory factor 5, MRC1 (CD206)] in the PDAC tumor microenvironment. Spatial point pattern analyses were conducted to assess the degree of colocalization between tumor cells and immune cells. Multivariable-adjusted Cox proportional hazards regression was used to assess associations with patient outcomes. In a multi-institutional cohort of 305 primary PDAC resection specimens, myeloid cells were abundant, enriched within stromal regions, highly heterogeneous across tumors, and differed by somatic genotype. High densities of CD15+ARG1+ immunosuppressive granulocytic cells and M2-polarized macrophages were associated with worse patient survival. Moreover, beyond cell density, closer proximity of M2-polarized macrophages to tumor cells was strongly associated with disease-free survival, revealing the clinical significance and biologic importance of immune cell localization within tumor areas. A diverse set of myeloid cells are present within the PDAC tumor microenvironment and are distributed heterogeneously across patient tumors. Not only the densities but also the spatial locations of myeloid immune cells are associated with patient outcomes, highlighting the potential role of spatially resolved myeloid cell subtypes as quantitative biomarkers for PDAC prognosis and therapy.


Predominance of PCR-ribotypes, 018 (smz) and 369 (trf) of Clostridium difficile in Japan: a potential relationship with other global circulating strains?
Global spread and evolutionary links of an epidemic Clostridium difficile strain (PCR-ribotype 027) have been noted in recent decades. However, in Japan, no outbreaks caused by type 027 have been reported to date. A total of 120 C. difficile isolates from patients at 15 hospitals during non-outbreak seasons between 2011 and 2013 as well as 18 and 21 isolates collected from two hospitals in 2010 and 2009, respectively, in outbreak periods in Japan, were examined. Among these 120 isolates, Japan-ribotypes smz and ysmz (subtype variant of smz) were the most predominant (39.2 %) followed by Japan-ribotype trf (15.8 %). Types smz/ysmz and trf were also concurrently predominant at two hospitals in the outbreak settings. Out of the five binary toxin-positive isolates observed, only one was PCR-ribotype 027 and another PCR-ribotype 078. Type smz was later found to correspond to PCR-ribotype 018. High rates of resistance against gatifloxacin, moxifloxacin, erythromycin and clindamycin were observed in the PCR-ribotype 018 isolates. Interestingly, all trf isolates were toxin A-negative, toxin B-positive, but they did not correspond to PCR-ribotype 017, thus being assigned a new ribotype (PCR-ribotype 369). In conclusion, PCR-ribotypes 018 (smz) and 369 (trf) were identified as major circulating strains in both outbreak and non-outbreak settings in Japan. Given their epidemiological relevance, molecular investigations are warranted to clarify potential evolutionary links with related strains found elsewhere, such as PCR-ribotypes 018 and 017 from Europe and North America.


Treatment of Cervical Myelopathy: Long-term Outcomes of Arthroplasty for Myelopathy Versus Radiculopathy, And Arthroplasty Versus Arthrodesis for Myelopathy.
Analysis of 2- and 7-year outcomes from a clinical trial comparing 2-level cervical disk arthroplasty (CDA) to anterior cervical discectomy and fusion (ACDF) in 287 patients with radiculopathy alone, and 110 patients with myelopathy alone or myelopathy with radiculopathy. To compare the long-term safety and effectiveness of CDA for myelopathy versus radiculopathy. CDA for myelopathy is safe and effective in short term. We analyzed Neck Disability Index (NDI), neck/arm pain, SF-36, neurological status, adverse events (AEs), and secondary surgeries at index and adjacent levels. All groups improved significantly for NDI, neck/arm pain, and physical component summary (PCS) scores from preoperative to postoperative. CDA Myelopathy versus CDA Radiculopathy: 2- and 7-year improvements were not significantly different. The 7-year score improvements for CDA Myelopathy and CDA Radiculopathy were: NDI (37.8 vs. 35.8, P=0.352), neck pain (12.0 vs. 12.1, P=0.477), arm pain (11.6 vs. 9.6, P=0.480), and PCS (14.1 vs. 13.7, P=0.863). The 2 groups had similar proportions of patients who maintained or improved their neurological status (87.2% vs. 93.5%, P=0.218), similar rates of serious AEs (54.5% vs. 57.5%, P=0.291) and similar rates of secondary surgeries at index (3.7% vs. 4.4%, P=0.839) and adjacent levels (3.7% vs. 7.6%, P=0.367). CDA Myelopathy versus ACDF myelopathy: 2 and 7-year improvements were not significantly different. The 7-year CDA and ACDF score improvements were: NDI (37.8 vs. 31.1, P=0.147), neck pain (12.0 vs. 10.4, P=0.337), arm pain (11.6 vs. 11.4, P=0.791), and PCS (14.1 vs. 11.2, P=0.363). The 2 groups had statistically similar proportions who maintained or improved their neurological status (87.2% vs. 96.2%, P=0.409), statistically similar overall rates of secondary surgeries at the index levels (3.7% vs. 9.4%, P=0.374), and statistically similar rates of secondary surgeries at adjacent levels (3.7% vs. 15.4%, P=0.088). CDA group demonstrated lower rates of serious AEs than ACDF (54.5% vs. 65.9%, P=0.019). CDA for myelopathy is a safe and effective long-term treatment.


Kinetics of elementary steps in the reactions of atomic bromine with isoprene and 1,3-butadiene under atmospheric conditions.
Laser flash photolysis of CF(2)Br(2) has been coupled with time-resolved detection of atomic bromine by resonance fluorescence spectroscopy to investigate the gas-phase kinetics of early elementary steps in the Br-initiated oxidations of isoprene (2-methyl-1,3-butadiene, Iso) and 1,3-butadiene (Bu) under atmospheric conditions. At T ≥ 526 K, measured rate coefficients for Br + isoprene are independent of pressure, suggesting that hydrogen transfer (1a) is the dominant reaction pathway. The following Arrhenius expression adequately describes all kinetic data at 526 K ≤ T ≤ 673 K: k(1a)(T) = (1.22 ± 0.57) × 10(-11) exp[(-2100 ± 280)/T] cm(3) molecule(-1) s(-1) (uncertainties are 2σ and represent precision of the Arrhenius parameters). At 271 K ≤ T ≤ 357 K, kinetic evidence for the reversible addition reactions Br + Iso ↔ Br-Iso (k(1b), k(-1b)) and Br + Bu ↔ Br-Bu (k(3b), k(-3b)) is observed. Analysis of the approach to equilibrium data allows the temperature- and pressure-dependent rate coefficients k(1b), k(-1b), k(3b), and k(-3b) to be evaluated. At atmospheric pressure, addition of Br to each conjugated diene occurs with a near-gas-kinetic rate coefficient. Equilibrium constants for the addition/dissociation reactions are obtained from k(1b)/k(-1b) and k(3b)/k(-3b), respectively. Combining the experimental equilibrium data with electronic structure calculations allows both second- and third-law analyses of thermochemistry to be carried out. The following thermochemical parameters for the addition reactions 1b and 3b at 0 and 298 K are obtained (units are kJ mol(-1) for Δ(r)H and J mol(-1) K(-1) for Δ(r)S; uncertainties are accuracy estimates at the 95% confidence level): Δ(r)H(0)(1b) = -66.6 ± 7.1, Δ(r)H(298)(1b) = -67.5 ± 6.6, and Δ(r)S(298)(3b) = -93 ± 16; Δ(r)H(0)(3b) = -62.4 ± 9.0, Δ(r)H(298)(3b) = -64.5 ± 8.5, and Δ(r)S(298)(3b) = -94 ± 20. Examination of the effect of added O(2) on Br kinetics under conditions where reversible adduct formation is observed allows the rate coefficients for the Br-Iso + O(2) (k(2)) and Br-Bu + O(2) (k(4)) reactions to be determined. At 298 K, we find that k(2) = (3.2 ± 1.0) × 10(-13) cm(3) molecule(-1) s(-1) independent of pressure (uncertainty is 2σ, precision only; pressure range is 25-700 Torr) whereas k(4) increases from 3.2 to 4.7 × 10(-13) cm(3) molecule(-1) s(-1) as the pressure increases from 25 to 700 Torr. Our results suggest that under atmospheric conditions, Br-Iso and Br-Bu react with O(2) to produce peroxy radicals considerably more rapidly than they undergo unimolecular decomposition. Hence, the very fast addition reactions appear to control the rates of Br-initiated formation of Br-Iso-OO and Br-Bu-OO radicals under atmospheric conditions. The peroxy radicals are relatively weakly bound, so conjugated diene regeneration via unimolecular decomposition reactions, though unimportant on the time scale of the reported experiments (milliseconds), is likely to compete effectively with bimolecular reactions of peroxy radicals under relatively warm atmospheric conditions as well as in 298 K competitive kinetics experiments carried out in large chambers.


Clonal integration supports the expansion from terrestrial to aquatic environments of the amphibious stoloniferous herb Alternanthera philoxeroides.
Effects of clonal integration on land plants have been extensively studied, but little is known about the role in amphibious plants that expand from terrestrial to aquatic conditions. We simulated expansion from terrestrial to aquatic habitats in the amphibious stoloniferous alien invasive alligator weed (Alternanthera philoxeroides) by growing basal ramets of clonal fragments in soils connected (allowing integration) or disconnected (preventing integration) to the apical ramets of the same fragments submerged in water to a depth of 0, 5, 10 or 15 cm. Clonal integration significantly increased growth and clonal reproduction of the apical ramets, but decreased both of these characteristics in basal ramets. Consequently, integration did not affect the performance of whole clonal fragments. We propose that alligator weed possesses a double-edged mechanism during population expansion: apical ramets in aquatic habitats can increase growth through connected basal parts in terrestrial habitats; however, once stolon connections with apical ramets are lost by external disturbance, the basal ramets in terrestrial habitats increase stolon and ramet production for rapid spreading. This may contribute greatly to the invasiveness of alligator weed and also make it very adaptable to habitats with heavy disturbance and/or highly heterogeneous resource supply.


Three-dimensional scroll waves of cAMP could direct cell movement and gene expression in Dictyostelium slugs.
Complex three-dimensional waves of excitation can explain the observed cell movement pattern in Dictyostelium slugs. Here we show that these three-dimensional waves can be produced by a realistic model for the cAMP relay system [Martiel, J. L. & Goldbeter, A. (1987) Biophys J. 52, 807-828]. The conversion of scroll waves in the prestalk zone of the slug into planar wave fronts in the prespore zone can result from a smaller fraction of relaying cells in the prespore zone. Further, we show that the cAMP concentrations to which cells in a slug are exposed over time display a simple pattern, despite the complex spatial geometry of the waves. This cAMP distribution agrees well with observed patterns of cAMP-regulated cell type-specific gene expression. The core of the spiral, which is a region of low cAMP concentration, might direct expression of stalk-specific genes during culmination.


Transitions to supported environments in England and Wales among elderly widowed and divorced women: the changing balance between co-residence with family and institutional care.
In this article we examine changes in the proportion of older widowed and divorced women in England and Wales moving from 'independent' to two kinds of 'supported' household--supported private households and institutions--during the decades 1971-81 and 1981-91. Our main aim was to see whether observed increases in institutionalisation over this period were the result of a decreased propensity to move to the households of relatives. We used the ONS Longitudinal Study, a record linkage study including individual level data from the 1971, 1981 and 1991 censuses of England and Wales. A multinomial logit model was used to investigate the correlates of transitions from independent to supported private households versus institutions among elderly widowed and divorced women. While the overall rate of transitions to join either supported private households or institutions was largely the same in the two decades, the balance between the two shifted markedly in favour of transitions to institutions. In terms of the limited range of covariates it was possible to consider, owner-occupiers were significantly more likely than tenants to move to supported private households than to institutions.


Beyond BOLD: optimizing functional imaging in stroke populations.
Blood oxygenation level-dependent (BOLD) signal changes are often assumed to directly reflect neural activity changes. Yet the real relationship is indirect, reliant on numerous assumptions, and subject to several sources of noise. Deviations from the core assumptions of BOLD contrast functional magnetic resonance imaging (fMRI), and their implications, have been well characterized in healthy populations, but are frequently neglected in stroke populations. In addition to conspicuous local structural and vascular changes after stroke, there are many less obvious challenges in the imaging of stroke populations. Perilesional ischemic changes, remodeling in regions distant to lesion sites, and diffuse perfusion changes all complicate interpretation of BOLD signal changes in standard fMRI protocols. Most stroke patients are also older than the young populations on which assumptions of neurovascular coupling and the typical analysis pipelines are based. We present a review of the evidence to show that the basic assumption of neurovascular coupling on which BOLD-fMRI relies does not capture the complex changes arising from stroke, both pathological and recovery related. As a result, estimating neural activity using the canonical hemodynamic response function is inappropriate in a number of contexts. We review methods designed to better estimate neural activity in stroke populations. One promising alternative to event-related fMRI is a resting-state-derived functional connectivity approach. Resting-state fMRI is well suited to stroke populations because it makes no performance demands on patients and is capable of revealing network-based pathology beyond the lesion site.


Selectivity of solid phase extraction of freshwater dissolved organic matter and its effect on ultrahigh resolution mass spectra.
Solid phase extraction (SPE) is often used for enrichment and clean-up prior to analysis of dissolved organic matter (DOM) by electrospray ionization (ESI) coupled to ultrahigh resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). It is generally accepted that extraction by SPE is not quantitative with respect to carbon concentration. However, little information is available on the selectivity of different SPE sorbents and the resulting effect for the acquired DOM mass spectra. Freshwater samples were extracted by the widely used PPL, HLB and C18 sorbents and the molecular composition and size distribution of the DOM in the extracts and in the permeates was compared to the original sample. Dissolved organic carbon (DOC) recoveries ranged between 20% and 65% for the three tested SPE sorbents. Size-exclusion chromatography coupled to organic carbon detection (SEC-OCD) revealed that limited recovery by PPL and HLB was primarily due to incomplete elution of a fraction of apparent high molecular weight from the solid phase. In contrast, incomplete retention on the solid phase, mainly observed for the C18 cartridge, was attributed to a fraction of low molecular weight. The FT-ICR mass spectra of the original sample and the SPE extracts did not differ significantly in their molecular weight distribution, but they showed sorbent specific differences in the degree of oxygenation and saturation. We concluded that the selective enrichment of freshwater DOM by SPE is less critical for subsequent FT-ICR MS analysis, because those fractions that are not sufficiently recovered have comparatively small effects on the mass spectra. This was confirmed by the extraction of model compounds, showing that very polar and small molecules are poorly extracted, but also have a low response in ESI-MS. Of the three tested SPE cartridges the PPL material offered the best properties for DOM enrichment for subsequent FT-ICR MS analysis as it minimizes too strong and too weak DOM-sorbent interactions.


Alternative energy efficient membrane bioreactor using reciprocating submerged membrane.
A novel membrane bioreactor (MBR) pilot system, using membrane reciprocation instead of air scouring, was operated at constant high flux and daily fluctuating flux to demonstrate its application under peak and diurnal flow conditions. Low and stable transmembrane pressure was achieved at 40 l/m(2)/h (LMH) by use of repetitive membrane reciprocation. The results reveal that the inertial forces acting on the membrane fibers effectively propel foulants from the membrane surface. Reciprocation of the hollow fiber membrane is beneficial for the constant removal of solids that may build up on the membrane surface and inside the membrane bundle. The membrane reciprocation in the reciprocating MBR pilot consumed less energy than coarse air scouring used in conventional MBR systems. Specific energy consumption for the membrane reciprocation was 0.072 kWh/m(3) permeate produced at 40 LMH flux, which is 75% less than for a conventional air scouring system as reported in literature without consideration of energy consumption for biological aeration (0.29 kWh/m(3)). The daily fluctuating flux test confirmed that the membrane reciprocation is effective to handle fluctuating flux up to 50 LMH. The pilot-scale reciprocating MBR system successfully demonstrated that fouling can be controlled via 0.43 Hz membrane reciprocation with 44 mm or higher amplitude.


Sympatric Atlantic puffins and razorbills show contrasting responses to adverse marine conditions during winter foraging within the North Sea.
Natural environments are dynamic systems with conditions varying across years. Higher trophic level consumers may respond to changes in the distribution and quality of available prey by moving to locate new resources or by switching diets. In order to persist, sympatric species with similar ecological niches may show contrasting foraging responses to changes in environmental conditions. However, in marine environments this assertion remains largely untested for highly mobile predators outside the breeding season because of the challenges of quantifying foraging location and trophic position under contrasting conditions. Differences in overwinter survival rates of two populations of North Sea seabirds (Atlantic puffins (Fratercula arctica) and razorbills (Alca torda)) indicated that environmental conditions differed between 2007/08 (low survival and thus poor conditions) and 2014/15 (higher survival, favourable conditions). We used a combination of bird-borne data loggers and stable isotope analyses to test 1) whether these sympatric species showed consistent responses with respect to foraging location and trophic position to these contrasting winter conditions during periods when body and cheek feathers were being grown (moult) and 2) whether any observed changes in moult locations and diet could be related to the abundance and distribution of potential prey species of differing energetic quality. Puffins and razorbills showed divergent foraging responses to contrasting winter conditions. Puffins foraging in the North Sea used broadly similar foraging locations during moult in both winters. However, puffin diet significantly differed, with a lower average trophic position in the winter characterised by lower survival rates. By contrast, razorbills' trophic position increased in the poor survival winter and the population foraged in more distant southerly waters of the North Sea. Populations of North Sea puffins and razorbills showed contrasting foraging responses when environmental conditions, as indicated by overwinter survival differed. Conservation of mobile predators, many of which are in sharp decline, may benefit from dynamic spatial based management approaches focusing on behavioural changes in response to changing environmental conditions, particularly during life history stages associated with increased mortality.


T cell epitope identification for bovine vaccines: an epitope mapping method for BoLA A-11.
T cell responses play an important role in immunity to parasites and other microbial agents of infectious diseases, therefore a number of T cell-directed vaccines are in development. Computer-driven algorithms that facilitate the discovery of T cell epitopes from protein and genome sequences are now being used to accelerate preclinical studies of human vaccines. Similar tools are not yet available for predicting T cell epitopes for animal vaccines, but there may be sufficient data available to begin the process of compiling the algorithms. We describe the construction of a novel mathematical 'matrix' that describes the properties of bovine major histocompatibility complex (BoLA) system antigen (BoLA) A-11 peptide ligands, developed for use with EpiMatrix, an existing T cell epitope-mapping algorithm. An alternative means of developing BoLA matrices, using the pocket profile method, is also discussed. Matrices such as the one described here may be used to develop T cell epitope-mapping tools for cattle and other ruminants. Epitope-mapping algorithms offer a significant advantage over other methods of epitope selection, such as the screening of synthetic overlapping peptides, because high throughput screening can be performed in silico, followed by ex vivo confirmatory studies. Furthermore, using epitope-mapping algorithms, putative T cell epitopes can be derived directly from genomic sequences, allowing researchers to circumvent labor-intensive cloning steps in the genome-to-vaccine discovery pathway.


Chlamydia pneumoniae infection in asthmatic patients.
This study was conducted on 30 asthmatic patients and 10 healthy controls. They were subjected to complete history talking, thorough clinical examination, and assessment of their ventilatory functions before and after bronchodilator. Two specimens were obtained from each patients, bronchoalveolar lavage and serum samples. Chlamydia pneumoniae antigens and antibodies (IgG and IgM) were detected using microimmunoflourescence technique. Samples giving positive results for C. pneumoniae antigen were examined for apoptosis. A significant correlation between asthma and C. pnumoniae was found especially in moderate and severe, long standing, steroid dependent asthma. In addition there was a significant difference between cases and controls regarding specific IgG and IgM. An association of C. pneumoniae with forced expiratory volume 1 (FEVI) was found. In addition C. pneumoniae was found to induce inhibition of programmed cell death. On conclusion, C. pneumoniae infection is associated with severe long standing asthma, and may be an important factor in acute exacerbation of asthma.


Narrow-band ultraviolet B treatment for vitiligo, pruritus, and inflammatory dermatoses.
Narrow-band ultraviolet B (NB-UVB) therapy has been used successfully for the treatment of inflammatory and pigmentary skin disorders including atopic dermatitis, psoriasis, mycosis fungoides, polymorphous light eruption, and vitiligo. This is a retrospective review of the treatment outcomes of 117 consecutive patients with vitiligo, pruritus, and other inflammatory dermatoses, excluding those with psoriasis and CTCL, who were treated with NB-UVB between 1998 and 2001 at our institution. Approximately 80% of all patients showed improvement in their condition. NB-UVB phototherapy was well tolerated, with no serious adverse effects. In patients with vitiligo, 6.4% had an abnormal thyroid-stimulating hormone level and 6.5% had anemia. NB-UVB may be considered as a viable therapeutic option in the treatment of vitiligo, pruritus, and other inflammatory dermatoses. Long-term adverse effects and cost-benefit analysis of NB-UVB therapy compared to other treatment modalities remain to be determined.


A Single-blinded, Randomized Clinical Trial of How to Implement an Evidence-based Treatment for Generalized Anxiety Disorder [IMPLEMENT]--Effects of Three Different Strategies of Implementation.
Despite long-standing calls to disseminate evidence-based treatments for generalized anxiety (GAD), modest progress has been made in the study of how such treatments should be implemented. The primary objective of this study was to test three competing strategies on how to implement a cognitive behavioral treatment (CBT) for out-patients with GAD (i.e., comparison of one compensation vs. two capitalization models). For our three-arm, single-blinded, randomized controlled trial (implementation of CBT for GAD [IMPLEMENT]), we recruited adults with GAD using advertisements in high-circulation newspapers to participate in a 14-session cognitive behavioral treatment (Mastery of your Anxiety and Worry, MAW-packet). We randomly assigned eligible patients using a full randomization procedure (1:1:1) to three different conditions of implementation: adherence priming (compensation model), which had a systematized focus on patients' individual GAD symptoms and how to compensate for these symptoms within the MAW-packet, and resource priming and supportive resource priming (capitalization model), which had systematized focuses on patients' strengths and abilities and how these strengths can be capitalized within the same packet. In the intention-to-treat population an outcome composite of primary and secondary symptoms-related self-report questionnaires was analyzed based on a hierarchical linear growth model from intake to 6-month follow-up assessment. This trial is registered at ClinicalTrials.gov (identifier: NCT02039193) and is closed to new participants. From June 2012 to Nov. 2014, from 411 participants that were screened, 57 eligible participants were recruited and randomly assigned to three conditions. Forty-nine patients (86%) provided outcome data at post-assessment (14% dropout rate). All three conditions showed a highly significant reduction of symptoms over time. However, compared with the adherence priming condition, both resource priming conditions indicated faster symptom reduction. The observer ratings of a sub-sample of recorded videos (n = 100) showed that the therapists in the resource priming conditions conducted more strength-oriented interventions in comparison with the adherence priming condition. No patients died or attempted suicide. To our knowledge, this is the first trial that focuses on capitalization and compensation models during the implementation of one prescriptive treatment packet for GAD. We have shown that GAD related symptoms were significantly faster reduced by the resource priming conditions, although the limitations of our study included a well-educated population. If replicated, our results suggest that therapists who implement a mental health treatment for GAD might profit from a systematized focus on capitalization models. Swiss Science National Foundation (SNSF-Nr. PZ00P1_136937/1) awarded to CF.


Multiple café au lait spots in familial patients with MAP2K2 mutation.
Recent advances in genetic diagnostic technologies have made the classic disease nosology highly complicated. This situation is exemplified by rasopathies, among which neurofibromatosis type 1 and Noonan syndrome represent prototypic entities. The former condition is characterized by multiple café au lait spots and neurofibromas, while the latter is characterized by distinct facial features, webbed neck, congenital heart disease, and a short stature. On rare occasions, the features of both neurofibromatosis and Noonan syndrome co-exist within an individual; such patients are diagnosed as having neurofibromatosis-Noonan syndrome. Here, we report familial patients with multiple café au lait spots and Noonan syndrome-like facial features. A mutation analysis unexpectedly revealed a mutation in MAP2K2 in both the propositus and his mother. The propositus fulfilled the diagnostic criteria for neurofibromatosis type 1, but his mother did not. Their phenotype was not consistent with that of cardio-facio-cutaneous syndrome, which is classically known to be associated with MAP2K2 mutations. The mother of the propositus had cervical cancer at the age of 23 years, consistent with the oncogenic tendency associated with rasopathies. The phenotypic combination of multiple café au lait spots and Noonan syndrome-like facial features suggested a diagnosis of neurofibromatosis-Noonan syndrome. Whether this condition represents a discrete disease entity or a variable expression of neurofibromatosis type 1 has long been debated. The present observation suggests that some perturbation in the RAS/MAPK signaling cascade results in multiple café au lait spots, a key diagnostic phenotype of rasopathies, although the exact mechanism remains to be elucidated.


Vaping for weight control: Findings from a qualitative study.
Smokers have expressed concern about weight gain once they stop smoking and weight gain is a risk factor associated with smoking relapse. Nicotine in e-cigarettes, as well as vaping behaviour, may support smoking cessation by reducing weight gain. This study explored the factors that influence attitudes towards, and awareness of, e-cigarettes and weight control post smoking cessation. Qualitative study involving focus groups with adults in the UK (n = 58) who were either exclusive vapers or dual users. There was limited awareness and/or inclination to vape to prevent weight gain after stopping smoking. Reasons for this centred on: the health gains of stopping smoking outweighing any potential weight gain; a lack of understanding of the appetite supressing effects of nicotine; a belief that vaping could not suppress appetite like a cigarette and could result in craving for certain flavours; concerns about the longer-term effects of e-cigarettes on health and the ethics of promoting vaping as way to support smoking cessation by limiting weight gain, especially for young women. Participants in this study do not appear inclined to use e-cigarettes to prevent weight gain after smoking cessation. There is a lack of understanding about why nicotine might help prevent weight gain and a concern that e-cigarette flavours could provoke cravings and that vaping may be unsafe in the long-term.


Dysregulation of endoplasmic reticulum stress response in skin wounds in a streptozotocin-induced diabetes mouse model.
Dysfunction in key cellular organelles has been linked to diabetic complications. This study intended to investigate the alterations in the unfolded protein response (UPR), autophagy, and mitochondrial function, which are part of the endoplasmic reticulum (ER) stress response, in wound healing (WH) under diabetes conditions. WH mouse models were used to evaluate the UPR, autophagy, mitochondrial fusion, fission, and biogenesis as well as mitophagy in the skin of control and diabetic mice at baseline and 10 days after wounding. The autophagic flux in response to high-glucose conditions was also evaluated in keratinocyte and fibroblast cell cultures. WH was impaired in the diabetic mouse model, and we found that the UPR and autophagy pathways were activated in skin wounds of control mice and in the non-wounded skin of diabetic mice. Moreover, high-glucose conditions induced autophagy in the keratinocyte and fibroblast cell cultures. However, mitophagy did not change in the skin of diabetic mice or the wounded skin. In addition, mitochondrial fusion was activated in control but not in the skin wounds of diabetic mice, while mitochondrial biogenesis is downregulated in the skin of diabetic mice. In conclusion, the activation of the UPR, autophagy, and mitochondrial remodeling are crucial for a proper WH. These results suggest that the increase in ER stress and autophagy in the skin of diabetic mice at baseline significantly escalated to pathological levels after wounding, contributing to impaired WH in diabetes.


Differential drug class-specific metastatic effects following treatment with a panel of angiogenesis inhibitors.
Inhibiting angiogenesis has become an important therapeutic strategy for cancer treatment but, like other current targeted therapies, benefits experienced for late-stage cancers can be curtailed by inherent refractoriness or by acquired drug resistance, requiring a need for better mechanistic understanding of such effects. Numerous preclinical studies have demonstrated that VEGF pathway inhibitors suppress primary tumour growth and metastasis. However, it has been recently reported that short-term VEGF and VEGFR inhibition can paradoxically accelerate tumour invasiveness and metastasis in certain models. Here we comprehensively compare the effects of both antibody and small molecule receptor tyrosine kinase (RTK) inhibitors targeting the VEGF-VEGFR pathway, using short-term therapy in various mouse models of metastasis. Our findings demonstrate that antibody inhibition of VEGF pathway molecules does not promote metastasis, in contrast to selected small molecule RTK inhibitors at elevated-therapeutic drug dosages. In particular, a multi-targeted RTK inhibitor, sunitinib, which most profoundly potentiated metastasis, also increased lung vascular permeability and promoted tumour cell extravasation. Mechanistically, sunitinib, but not anti-VEGF treatment, attenuated endothelial barrier function in culture and caused a global inhibition of protein tyrosine phosphorylation, including molecules important for maintaining endothelial cell-cell junctions. Together these findings indicate that, rather than a specific consequence of inhibiting the VEGF signalling pathway, pharmacological inhibitors of the VEGF pathway can have dose- and drug class-dependent side-effects on the host vasculature. These findings also advocate for the continued identification of mechanisms of resistance to anti-angiogenics and for therapy development to overcome it.


Bayesian adaptive trial design for a newly validated surrogate endpoint.
The evaluation of surrogate endpoints for primary use in future clinical trials is an increasingly important research area, due to demands for more efficient trials coupled with recent regulatory acceptance of some surrogates as 'valid.' However, little consideration has been given to how a trial that utilizes a newly validated surrogate endpoint as its primary endpoint might be appropriately designed. We propose a novel Bayesian adaptive trial design that allows the new surrogate endpoint to play a dominant role in assessing the effect of an intervention, while remaining realistically cautious about its use. By incorporating multitrial historical information on the validated relationship between the surrogate and clinical endpoints, then subsequently evaluating accumulating data against this relationship as the new trial progresses, we adaptively guard against an erroneous assessment of treatment based upon a truly invalid surrogate. When the joint outcomes in the new trial seem plausible given similar historical trials, we proceed with the surrogate endpoint as the primary endpoint, and do so adaptively-perhaps stopping the trial for early success or inferiority of the experimental treatment, or for futility. Otherwise, we discard the surrogate and switch adaptive determinations to the original primary endpoint. We use simulation to test the operating characteristics of this new design compared to a standard O'Brien-Fleming approach, as well as the ability of our design to discriminate trustworthy from untrustworthy surrogates in hypothetical future trials. Furthermore, we investigate possible benefits using patient-level data from 18 adjuvant therapy trials in colon cancer, where disease-free survival is considered a newly validated surrogate endpoint for overall survival.