Functional characterization of a FUS mutant zebrafish line as a novel genetic model for ALS.
Mutations in Fused in sarcoma (FUS), an RNA-binding protein, are known to cause Amyotrophic Lateral Sclerosis (ALS). However, molecular mechanisms due to loss of FUS function remain unclear and controversial. Here, we report the characterization and phenotypic analysis of a deletion mutant of the unique FUS orthologue in zebrafish where Fus protein levels are depleted. The homozygous mutants displayed a reduced lifespan as well as impaired motor abilities associated with specific cellular deficits, including decreased motor neurons length and neuromuscular junctions (NMJ) fragmentation. Furthermore, we demonstrate that these cellular impairments are linked to the misregulation of mRNA expression of acetylcholine receptor (AChR) subunits and histone deacetylase 4, markers of denervation and reinnervation processes observed in ALS patients. In addition, fus loss of function alters tau transcripts favoring the expression of small tau isoforms. Overall, this new animal model extends our knowledge on FUS and supports the relevance of FUS loss of function in ALS physiopathology.


Modelling a turbidity current in Soyang reservoir (Korea) and its control using a selective withdrawal facility.
Persistent turbidity in reservoirs and their downstream after flood events is one of most important environmental issues in Korea. Recently, modification of withdrawal facility and installation of a new selective withdrawal structure (SWS) have been implemented for the mitigation of downstream impact and sediment loading into water treatment facilities. This study was to explore the characteristics of flood density flow induced into Soyang Reservoir and the transport processes of suspended sediments (SS) through application of coupled two-dimensional hydrodynamic and particle dynamic models (TM-1, TM-2 and TM-3). The TM-3 including a turbidity attenuation rate as a lumped parameter showed best performance in reproducing the magnitude and distribution of SS in the reservoir. The validated model was applied to evaluate the effectiveness of SWS, which was designed for the reservoir, with 6 different historical flood events. The magnitude of vertical mixing of the turbidity plume and its persistence within the reservoir were closely correlated to the ratio of the volume of turbidity flow to the total reservoir storage (the theta value). The operation of SWS showed a positive effect as long as theta is between 0.3 and 0.6 but negative when theta = 0.83 for the study reservoir, thus it should be optimized based on the theta value for a better management of the reservoir.


A 95-year-old woman with leucocytosis and eosinophilia: anaplastic carcinoma in an ectopic thyroid.
A 95-year-old woman had been treated over the past 8 years for progressive dysphagia. When her condition worsened, blood tests revealed the presence of leucocytosis and eosinophilia in the absence of anaemia or thrombocytopenia. Within 11 days of diagnosis, the patient died of respiratory failure. Necropsy showed normal thyroid tissue and an absence of infectious disease. However, an upper mediastinal tumour was found and was histopathologically diagnosed as a neoplastic transformation of the ectopic thyroid. Only 1% of endothoracic goitres present as ectopic or autonomous goitre with no parenchymal or vascular connection to the thyroid gland. This case represents a very rare situation in which a leukemoid reaction and peripheral hypereosinophilia were observed as a manifestation of an anaplastic thyroid carcinoma in an ectopic mass.


ePlant: Visualizing and Exploring Multiple Levels of Data for Hypothesis Generation in Plant Biology.
A big challenge in current systems biology research arises when different types of data must be accessed from separate sources and visualized using separate tools. The high cognitive load required to navigate such a workflow is detrimental to hypothesis generation. Accordingly, there is a need for a robust research platform that incorporates all data and provides integrated search, analysis, and visualization features through a single portal. Here, we present ePlant (http://bar.utoronto.ca/eplant), a visual analytic tool for exploring multiple levels of Arabidopsis thaliana data through a zoomable user interface. ePlant connects to several publicly available web services to download genome, proteome, interactome, transcriptome, and 3D molecular structure data for one or more genes or gene products of interest. Data are displayed with a set of visualization tools that are presented using a conceptual hierarchy from big to small, and many of the tools combine information from more than one data type. We describe the development of ePlant in this article and present several examples illustrating its integrative features for hypothesis generation. We also describe the process of deploying ePlant as an "app" on Araport. Building on readily available web services, the code for ePlant is freely available for any other biological species research.


Interaction of Salmonella choleraesuis, Salmonella dublin and Salmonella typhimurium with porcine and bovine terminal ileum in vivo.
Quantitative experiments on the interaction of Salmonella choleraesuis and Salmonella dublin with porcine and bovine intestinal epithelia yielded no evidence to suggest that host restriction of S. choleraesuis and S. dublin for pigs and calves respectively could be explained in terms of the patterns of intestinal invasion observed in ligated ileal loops in vivo, at 3 h after challenge. No evidence was found to support the idea that Peyer's patches, or specifically M cells, are the major route of entry for these serotypes in vivo. Three hours after loop inoculation, each serotype was recovered in comparable numbers from either absorptive or Peyer's patch mucosae present in the same ileal loop, indicating that both types of tissue are involved in the early stages of the enteropathogenic process induced by both serotypes. More detailed transmission electron microscopic (TEM) analyses of follicle-associated epithelia (FAE) challenged with S. choleraesuis showed that in the same region of FAE, organisms invaded both M cells and enterocytes directly; comparable detailed TEM studies with S. dublin could not be carried out because of the tissue-destructive properties of this serotype. S. dublin was clearly more histotoxic than S. choleraesuis as had previously been found in rabbits: this difference is almost certainly due to a tissue-damaging toxin which is neither host nor gut-tissue specific. The tissue-destructive potential of S. dublin has profound implications for the measurement of and the assignment of significance to the invasiveness of S. dublin. S. dublin was nearly always seen entering gut cells in micro-colonies whereas S. choleraesuis entered mainly as single organisms or small groups of two or three.


Phenotypes of COPD in an Austrian population : National data from the POPE study.
Chronic obstructive pulmonary disease (COPD) represents a major global health problem; however, there are no data regarding clinical phenotypes of these patients in Austria. This was an analysis from the Austrian cohort of the cross-sectional Phenotypes of COPD in Central and Eastern Europe (POPE) study, which was offered to patients with stable COPD in a real-life setting. Patients were recruited at 5 different outpatient facilities in 3 different provinces in Austria. All consecutive patients aged ≥40 years with a diagnosis of COPD confirmed by a post-bronchodilator forced expired volume in 1 s/forced vital capacity (FEV1/FVC) ratio <0.7 during a stable state (≥4 weeks without exacerbation or worsening of any relevant comorbidities) were considered eligible. The primary aim of this study was to assess the prevalence of phenotypes according to predefined criteria. Secondary aims included analyses of differences in patient characteristics, symptom load, comorbidities, and pharmacological treatment. Among 283 patients fulfilling the inclusion criteria, 49.5% were considered non-exacerbators, 21.6% were classified as exacerbators with chronic bronchitis, 21.2% exacerbators without chronic bronchitis, and 7.8% were patients with an asthma-COPD overlap. Exacerbators had significantly higher prevalence of symptoms, lower lung function and exercise capacity, and a higher prevalence of comorbidities, such as heart failure and depression, compared with the other patient phenotypes. A large majority of patients with stable COPD in this cohort received inhaled triple therapy, irrespective of exacerbation history. There were significant differences in COPD outcome measures between predefined phenotypes of COPD in this study. The majority of patients with stable COPD in this Austrian population were not treated according to current COPD guidelines. While non-exacerbators appear to have been overtreated, patients with an asthma-COPD overlap appear to have been undertreated.


Glycine receptors in the retinas of normal and spastic mutant mice.
Spastic mutant mice have abnormal gait and righting behavior, and the responses of their retinal ganglion cells have recently been shown to be abnormal. The former defects have been linked to a reduction of glycine-receptor density in the spinal cord of spastic mutants, but the cause of the retinal defects has not yet been determined. The authors thus tested for reduced glycine-receptor density in the mutant retina by comparing the levels of glycine receptors in the retinas of spastic mutant mice with those found in normal mice. Indirect immunofluorescence histochemistry was employed, using monoclonal antibodies directed against the alpha- and beta-subunits of the receptor and against the 93-kd cytoplasmic receptor-associated protein, gephyrin. In normal mice, all glycine-receptor antibodies labeled two laminae of the inner plexiform layer (IPL): a broad band in the distal third of the IPL and a narrow band in the middle of the IPL. Lighter labeling was also seen in the outer plexiform layer with these antibodies. In spastic mutant mice, the glycine-receptor labeling of the IPL was reduced markedly. However, the overall structure of the spastic mutant retina was not disrupted because the distribution and intensity of both a presynaptic marker (synaptophysin) and a marker for the rod bipolar cell (protein kinase C) in the mutant retina were indistinguishable from those in normal retinas. The glycine-receptor distribution in normal mice was consistent with that previously reported for the rat and with the distribution of glycine responsiveness of dissociated rodent bipolar cells. The reduced levels of glycine receptors in spastic mice help explain the abnormal ganglion cell responses in the spastic mutant.


Microscopic visualization of Wuchereria and Brugia larval stages in intact cleared mosquitoes.
Over the past several decades, epidemiologic data from filarial vectors typically has been obtained by mass dissection or by dissection of individual specimens. The former is quick and easy to do on large numbers of insects but provides no information on the frequency distribution of infection, presence of early developmental stages, or larval location; the latter is labor-intensive and tedious. We describe a new technique that can provide data comparable to those obtained by individual dissection, including calculation of infection and infective rates, and this technique is easy enough to accommodate large numbers of insects. Brief treatment of ethanol-fixed, intact mosquitoes in sodium hypochlorite, followed by treatments in increasing concentrations of ethanol and an organic solvent allowed microscopic visualization of filarial larvae within the abdomen, thorax, head, and proboscis of Brugia malayi-infected Aedes aegypti and Wuchereria bancrofti-infected Anopheles punctulatus. We compared the classic techniques to our technique using Ae. aegypti infected by feeding on jirds with B. malayi microfilaremias. Comparisons of the infective rate, total number of infective stage larvae (L3s) observed, and locations of L3s showed that this new technique was comparable to the established methods, while being faster and more precise in determining the location of larvae.


Deletion of the monkeypox virus inhibitor of complement enzymes locus impacts the adaptive immune response to monkeypox virus in a nonhuman primate model of infection.
Monkeypox virus (MPXV) is an orthopoxvirus closely related to variola virus, the causative agent of smallpox. Human MPXV infection results in a disease that is similar to smallpox and can also be fatal. Two clades of MPXV have been identified, with viruses of the central African clade displaying more pathogenic properties than those within the west African clade. The monkeypox inhibitor of complement enzymes (MOPICE), which is not expressed by viruses of the west African clade, has been hypothesized to be a main virulence factor responsible for increased pathogenic properties of central African strains of MPXV. To gain a better understanding of the role of MOPICE during MPXV-mediated disease, we compared the host adaptive immune response and disease severity following intrabronchial infection with MPXV-Zaire (n = 4), or a recombinant MPXV-Zaire (n = 4) lacking expression of MOPICE in rhesus macaques (RM). Data presented here demonstrate that infection of RM with MPXV leads to significant viral replication in the peripheral blood and lungs and results in the induction of a robust and sustained adaptive immune response against the virus. More importantly, we show that the loss of MOPICE expression results in enhanced viral replication in vivo, as well as a dampened adaptive immune response against MPXV. Taken together, these findings suggest that MOPICE modulates the anti-MPXV immune response and that this protein is not the sole virulence factor of the central African clade of MPXV.


Histone H2AFX Links Meiotic Chromosome Asynapsis to Prophase I Oocyte Loss in Mammals.
Chromosome abnormalities are common in the human population, causing germ cell loss at meiotic prophase I and infertility. The mechanisms driving this loss are unknown, but persistent meiotic DNA damage and asynapsis may be triggers. Here we investigate the contribution of these lesions to oocyte elimination in mice with chromosome abnormalities, e.g. Turner syndrome (XO) and translocations. We show that asynapsed chromosomes trigger oocyte elimination at diplonema, which is linked to the presence of phosphorylated H2AFX (γH2AFX). We find that DNA double-strand break (DSB) foci disappear on asynapsed chromosomes during pachynema, excluding persistent DNA damage as a likely cause, and demonstrating the existence in mammalian oocytes of a repair pathway for asynapsis-associated DNA DSBs. Importantly, deletion or point mutation of H2afx restores oocyte numbers in XO females to wild type (XX) levels. Unexpectedly, we find that asynapsed supernumerary chromosomes do not elicit prophase I loss, despite being enriched for γH2AFX and other checkpoint proteins. These results suggest that oocyte loss cannot be explained simply by asynapsis checkpoint models, but is related to the gene content of asynapsed chromosomes. A similar mechanistic basis for oocyte loss may operate in humans with chromosome abnormalities.


Cell Migration and Invadopodia Formation Require a Membrane-binding Domain of CARMIL2.
CARMILs regulate capping protein (CP), a critical determinant of actin assembly and actin-based cell motility. Vertebrates have three conserved CARMIL genes with distinct functions. In migrating cells, CARMIL2 is important for cell polarity, lamellipodial assembly, ruffling, and macropinocytosis. In cells, CARMIL2 localizes with a distinctive dual pattern to vimentin intermediate filaments and to membranes at leading edges and macropinosomes. The mechanism by which CARMIL2 localizes to membranes has not been defined. Here, we report that CARMIL2 has a conserved membrane-binding domain composed of basic and hydrophobic residues, which is necessary and sufficient for membrane localization, based on expression studies in cells and on direct binding of purified protein to lipids. Most important, we find that the membrane-binding domain is necessary for CARMIL2 to function in cells, based on rescue expression with a set of biochemically defined mutants. CARMIL1 and CARMIL3 contain similar membrane-binding domains, based on sequence analysis and on experiments, but other CPI motif proteins, such as CD2AP, do not. Based on these results, we propose a model in which the membrane-binding domain of CARMIL2 tethers this multidomain protein to the membrane, where it links dynamic vimentin filaments with regulation of actin assembly via CP.


Growth hormone predicts bone density in elderly women.
Evidence is accumulating that the risk of osteoporosis may be influenced by environmental factors during intrauterine and early postnatal life; such programming might be mediated through modification of the GH/IGF-1 axis during critical periods in its development. To address this issue, we explored the relationships among birth weight, circulating GH profile, bone density, and bone loss rate in a group of British women. The study population consisted of 38 women 60-75 years old resident in Hertfordshire for whom detailed birth records were available. Twenty-four-hour circulating GH profiles were obtained during an inpatient stay on a metabolic ward, after an overnight rest. The circulating profile of GH was characterised by estimating the peak, median, trough, and total concentrations from 72 samples measured sequentially over 24 h in each subject. Bone mineral density was assessed at the lumbar spine and femoral neck at baseline and at follow-up 4 years later. Lumbar spine bone mineral content (BMC) and density (BMD) were positively associated with all measures of GH concentration, although relationships were strongest for BMC with trough GH (r = 0.47, P < 0.01). Associations persisted after adjustment for age, body mass index, smoking, alcohol consumption, physical activity, and osteoarthritis score in multiple regression models. However, associations of GH concentration with femoral neck BMC were weak, and there was no association between any measure of GH concentration and bone loss at either site. Total (integrated) daily GH concentration tended to increase (P = 0.08) with rising birth weight, while IGF-1 concentration fell (P = 0.05) with rising birth weight, suggesting a role for the GH/IGF-1 axis in the programming of adult bone mass among women.


The next generation of iron fertilization experiments in the Southern Ocean.
Of the various macro-engineering schemes proposed to mitigate global warming, ocean iron fertilization (OIF) is one that could be started at short notice on relevant scales. It is based on the reasoning that adding trace amounts of iron to iron-limited phytoplankton of the Southern Ocean will lead to blooms, mass sinking of organic matter and ultimately sequestration of significant amounts of atmospheric carbon dioxide (CO2) in the deep sea and sediments. This iron hypothesis, proposed by John Martin in 1990 (Martin 1990 Paleoceanography 5, 1-13), has been tested by five mesoscale experiments that provided strong support for its first condition: stimulation of a diatom bloom accompanied by significant CO2 drawdown. Nevertheless, a number of arguments pertaining to the fate of bloom biomass, the ratio of iron added to carbon sequestered and various side effects of fertilization, continue to cast doubt on its efficacy. The idea is also unpopular with the public because it is perceived as meddling with nature. However, this apparent consensus against OIF is premature because none of the published experiments were specifically designed to test its second condition pertaining to the fate of iron-induced organic carbon. Furthermore, the arguments on side effects are based on worst-case scenarios. These doubts, formulated as hypotheses, need to be tested in the next generation of OIF experiments. We argue that such experiments, if carried out at appropriate scales and localities, will not only show whether the technique will work, but will also reveal a wealth of insights on the structure and functioning of pelagic ecosystems in general and the krill-based Southern Ocean ecosystem, in particular. The outcomes of current models on the efficacy and side effects of OIF differ widely, so data from adequately designed experiments are urgently needed for realistic parametrization. OIF is likely to boost zooplankton stocks, including krill, which could have a positive effect on recovery of the great whale populations. Negative effects of possible commercialization of OIF can be controlled by the establishment of an international body headed by scientists to supervise and monitor its implementation.


Plasmonic Detection of Glucose in Serum Based on Biocatalytic Shape-Altering of Gold Nanostars.
Nanoparticles have been used as signal transducers for optical readouts in biosensors. Optical approaches are cost-effective with easy readout formats for clinical diagnosis. We present a glucose biosensor based on the biocatalytic shape-altering of gold nanostars via silver deposition. Improved sensitivity was observed due to the nanostars clustering after being functionalised with glucose oxidase (GOx). The biosensor quantified glucose in the serum samples with a 1:1000 dilution factor, and colorimetrically distinguished between the concentrations. The assay demonstrated good specificity and sensitivity. The fabricated glucose biosensor is a rapid kinetic assay using a basic entry level laboratory spectrophotometric microplate reader. Such a biosensor could be very useful in resource-constrained regions without state-of-the-art laboratory equipment. Furthermore, naked eye detection of glucose makes this a suitable biosensor for technology transfer to other point-of-care devices.


Atherothrombosis and the NLRP3 inflammasome - endogenous mechanisms of inhibition.
Recently, the CANTOS (Canakinumab Anti-Inflammatory Thrombosis Outcomes Study) showed the successful anti-inflammatory benefit of canakinumab, a monoclonal antibody targeting interleukin-1ß (IL-1ß) toward major cardiovascular events (MACE) in patients with a previous myocardial infarction (MI). The magnitude of reduction in MACE was directly attributed to a reduction witnessed in IL-6 and C-reactive protein (CRP) and highlighted the therapeutic potential of selectively targeting IL-1ß for atherosclerotic disease, a notion previously introduced in animal models. IL-1ß is involved in the downstream activation of the IL-6 receptor, which itself has been previously implicated as a target for atherothrombosis from Mendelian randomization studies. Further support has been garnered with the results of CIRT (Cardiovascular Inflammation Reduction Trial), which showed the inability of low-dose methotrexate to reduce IL-1ß, IL-6, or high-sensitivity CRP (hsCRP) in addition to MACE among patients with prior MI or multivessel coronary artery disease (CAD) but with normal hsCRP levels. Therefore, elucidation of therapeutic targets against the IL-1ß pathway is of immense interest currently in treating atherothrombosis. Upstream and serving as an activator of IL-1ß lies the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome that has been well described in animal models to be activated by cholesterol crystals or hypoxia to promote cleavage and secretion of IL-1ß and IL-18 that lead to atherosclerotic deposition in arteries. Given the direct implication of an atherogenic role to the NLRP3 inflammasome in generating these cytokines, NLRP3 inhibitors are of interest with the consideration to move upstream from the initial success of anti-IL-1ß therapy. With further discussion of the existing knowledge on the proinflammatory relationship of the NLRP3 inflammasome with atherosclerosis, this review summarizes and critically evaluates the preclinical and interventional findings of endogenous NLRP3 inflammasome inhibition in attempts to elucidate anti-inflammatory mechanisms, and therapeutic targets against atherothrombosis. Further investigation focusing on the endogenous mechanisms of inhibition of the NLRP3 inflammasome would uncover diagnostic routes from defective means in inflammatory resolution. Specifically, pro-resolving lipid mediators, autophagy, and phosphorylation/dephosphorylation mechanisms are 3 points of worthy investigation from existing evidence.


Incidence of tuberculosis, hepatitis, brucellosis, and shigellosis in British medical laboratory workers.
A retrospective postal survey of 21 000 medical laboratory workers in England and Wales showed 18 new cases of pulmonary tuberculosis in 1971, a five-times increased risk of acquiring the disease compared with the general population. Technicians were at greatest risk, especially if they worked in morbid anatomy departments. Of the 35 cases of hepatitis, the technicians were again the occupational group most likely to acquire the disease. Microbiology staff were twice as likely to report shigellosis as those in other pathology divisions but only one case of brucellosis was reported in the whole laboratory population. A similar survey carried out in 1973 of 3000 Scottish medical laboratory workers corroborates the results from England and Wales. Medical laboratory workers continue to experience a considerable risk of developing an occupationally acquired infection. Improvements in staff safety and health care seem to be necessary.


Hyperthyroidism in dogs caused by consumption of thyroid-containing head meat.
Two female spayed dogs belonging to the same owner were admitted for further examinations because of clinical signs and laboratory values compatible with hyperthyroidism. Sonography of the ventral aspect of the neck revealed small thyroid glands in both dogs. The hypothesis that the dogs suffered from alimentary hyperthyroidism caused by feeding head meat containing thyroid gland tissue was confirmed by consultation of the slaughtering plant, determination of iodine concentrations in deep-frozen samples and hormone measurements in 5 other dogs receiving head meat from the same supplier. After changing the diet, thyroxine concentrations declined and clinical signs were no longer observed.


Structural and biochemical basis for polyamine binding to the Tp0655 lipoprotein of Treponema pallidum: putative role for Tp0655 (TpPotD) as a polyamine receptor.
Tp0655 of Treponema pallidum, the causative agent of syphilis, is predicted to be a 40 kDa membrane lipoprotein. Previous sequence analysis of Tp0655 noted its homology to polyamine-binding proteins of the bacterial PotD family, which serve as periplasmic ligand-binding proteins of ATP-binding-cassette (ABC) transport systems. Here, the 1.8 A crystal structure of Tp0655 demonstrated structural homology to Escherichia coli PotD and PotF. The latter two proteins preferentially bind spermidine and putrescine, respectively. All of these proteins contain two domains that sandwich the ligand between them. The ligand-binding site of Tp0655 can be occupied by 2-(N-morpholino)ethanesulfanoic acid, a component of the crystallization medium. To discern the polyamine binding preferences of Tp0655, the protein was subjected to isothermal titration calorimetric experiments. The titrations established that Tp0655 binds polyamines avidly, with a marked preference for putrescine (Kd=10 nM) over spermidine (Kd=430 nM), but the related compounds cadaverine and spermine did not bind. Structural comparisons and structure-based sequence analyses provide insights into how polyamine-binding proteins recognize their ligands. In particular, these comparisons allow the derivation of rules that may be used to predict the function of other members of the PotD family. The sequential, structural, and functional homology of Tp0655 to PotD and PotF prompt the conclusion that the former likely is the polyamine-binding component of an ABC-type polyamine transport system in T. pallidum. We thus rename Tp0655 as TpPotD. The ramifications of TpPotD as a polyamine-binding protein to the parasitic strategy of T. pallidum are discussed.


Connexin 40 promoter-based enrichment of embryonic stem cell-derived cardiovascular progenitor cells.
Pluripotent embryonic stem (ES) cells that can differentiate into functional cardiomyocytes as well as vascular cells in cell culture may open the door to cardiovascular cell transplantation. However, the percentage of ES cells in embryoid bodies (EBs) which spontaneously undergo cardiovascular differentiation is low (<10%), making strategies for their specific labeling and purification indispensable. The human connexin 40 (Cx40) promoter was isolated and cloned in the vector pEGFP. The specificity of the construct was initially assessed in Xenopus embryos injected with Cx40-EGFP plasmid DNA. Stable Cx40-EGFP ES cell clones were differentiated and fluorescent cells were enriched manually as well as via fluorescence-activated cell sorting. Characterization of these cells was performed with respect to spontaneous beating as well as via RT-PCRs and immunofluorescent stainings. Cx40-EGFP reporter plasmid injection led to EGFP fluorescence specifically in the abdominal aorta of frog tadpoles. After crude manual enrichment of highly Cx40-EGFP-positive EBs, the appearance of cardiac and vascular structures was increased approximately 3-fold. Immunofluorescent stainings showed EGFP expression exclusively in vascular-like structures simultaneously expressing von Willebrand factor and in formerly beating areas expressing alpha-actinin. Cx40-EGFP-expressing EBs revealed significantly higher numbers of beating cardiomyocytes and vascular-like structures. Semiquantitative RT-PCRs confirmed an enhanced cardiovascular differentiation as shown for the cardiac markers Nkx2.5 and MLC2v, as well as the endothelial marker vascular endothelial cadherin. Our work shows the feasibility of specific labeling and purification of cardiovascular progenitor cells from differentiating EBs based on the Cx40 promoter. We provide proof of principle that the deleted CD4 (DeltaCD4) surface marker-based method for magnetic cell sorting developed by our group will be ideally suitable for transference to this promoter.


Physiological and Molecular Function of the Sodium/Hydrogen Exchanger NHA2 (SLC9B2).
NHA2, also known as SLC9B2, is an orphan intracellular Na+/H+ exchanger (NHE) that has been associated with arterial hypertension and diabetes mellitus in humans. The objective of this NCCR TransCure project was to define the physiological and molecular function of NHA2, to develop a high resolution kinetic transport assay for NHA2 and to identify specific and potent compounds targeting NHA2. In this review, we summarize the results of this highly interdisciplinary and interfaculty effort, led by the groups of Proffs. Jean-Louis Reymond, Christoph von Ballmoos and Daniel Fuster.


Streptococcal reporter gene-fusion vector for identification of in vivo expressed genes.
To study streptococcal genes that are specifically induced in the host during endocarditis, we have developed a novel plasmid for use in in vivo expression technology (IVET). This IVET uses an integration plasmid, pAK36, that carries dual (amy-cat) reporter genes. A gene-fusion strain library was constructed with the plasmid randomly inserted into the chromosome of Streptococcus gordonii V288 by insertion-duplication. The library was inoculated intravenously into a rabbit that had been prepared for experimental endocarditis. Beginning 6 h after the inoculation, the rabbit was given chloramphenicol (Cm) intravenously twice a day to a final serum level of 5 microg/ml and was euthanized 3 days later. The aortic valve vegetations containing Cm(R) S. gordonii clones were cultured. Colonies were screened in vitro for negative amylase activity and sensitivity to Cm. Forty-eight such colonies showed 13 different insertion patterns when Southern hybridization blots were probed with labeled pAK36. For each of the 13 isolates, the gene fragment proximal to the insertion of the reporter amy-cat was cloned, and its nucleotide sequence was determined. Functions of these genes were inferred by their homology to known genes. Therefore, this novel IVET vector can be useful for identification of in vivo induced genes in S. gordonii and other streptococcal species.


A Food First Approach to Carbohydrate Supplementation in Endurance Exercise: A Systematic Review.
This systematic review analyzed whether carbohydrate source (food vs. supplement) influenced performance and gastrointestinal (GI) symptoms during endurance exercise. Medline, SPORTDiscus, and citations were searched from inception to July 2021. Inclusion criteria were healthy, active males and females aged >18 years, investigating endurance performance, and GI symptoms after ingestion of carbohydrate from a food or supplement, <60 min before or during endurance exercise. The van Rosendale scale was used to determine risk of bias, with seven studies having low risk of bias. A total of 151 participants from 15 studies were included in the review. Three studies provided 0.6-1 g carbohydrate/kg body mass during 5-45 min precycling exercise (duration 60-70 min) while 12 studies provided 24-80 g/hr carbohydrate during exercise (60-330 min). Except one study that suggested a likely harmful effect (magnitude-based inferences) of a bar compared to a gel consumed during exercise on cycling performance, there were no differences in running (n = 1) or cycling (n = 13) performance/capacity between food and supplemental sources. Greater GI symptoms were reported with food compared with supplemental sources. Highly heterogenous study designs for carbohydrate dose and timing, as well as exercise protocol and duration, make it difficult to compare findings between studies. A further limitation results from only one study assessing running performance. Food choices of carbohydrate consumed immediately before and during endurance exercise result in similar exercise performance/capacity responses to supplemental carbohydrate sources, but may slightly increase GI symptoms in some athletes, particularly with exercise >2 hr.


Genome rearrangement based on reversals that preserve conserved intervals.
The order of genes in the genomes of species can change during evolution and can provide information about their phylogenetic relationship. An interesting method to infer the phylogenetic relationship from the gene orders is to use different types of rearrangement operations and to find possible rearrangement scenarios using these operations. One of the most common rearrangement operations is reversals, which reverse the order of a subset of neighbored genes. In this paper, we study the problem to find the ancestral gene order for three species represented by their gene orders. The rearrangement scenario should use a minimal number of reversals and no other rearrangement operations. This problem is called the Median problem and is known to be NP-complete. In this paper, we describe a heuristic algorithm for finding solutions to the Median problem that searches for rearrangement scenarios with the additional property that gene groups should not be destroyed by reversal operations. The concept of conserved intervals for signed permutations is used to describe such gene groups. We show experimentally, for different types of test problems, that the proposed algorithm produces very good results compared to other algorithms for the Median problem. We also integrate our reversal selection procedure into the well-known MGR and GRAPPA algorithms and show that they achieve a significant speedup while obtaining solutions of the same quality as the original algorithms on the test problems.


Shifting tasks from pharmacy to non-pharmacy personnel for providing antiretroviral therapy to people living with HIV: a systematic review and meta-analysis.
Lay people or non-pharmacy health workers with training could dispense antiretroviral therapy (ART) in resource-constrained countries, freeing up time for pharmacists to focus on more technical tasks. We assessed the effectiveness of such task-shifting in low-income and middle-income countries. We conducted comprehensive searches of peer-reviewed and grey literature. Two authors independently screened search outputs, selected controlled trials, extracted data and resolved discrepancies by consensus. We performed random-effects meta-analysis and assessed certainty of evidence using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach. Three studies with 1993 participants met the inclusion criteria, including two cluster trials conducted in Kenya and Uganda and an individually randomised trial conducted in Brazil. We found very low certainty evidence regarding mortality due to the low number of events. Therefore, we are uncertain whether there is a true increase in mortality as the effect size suggests, or a reduction in mortality between pharmacy and non-pharmacy models of dispensing ART (risk ratio (RR) 1.86, 95% CI 0.44 to 7.95, n=1993, three trials, very low certainty evidence). There may be no differences between pharmacy and non-pharmacy models of dispensing ART on virological failure (risk ratio (RR) 0.92, 95% CI 0.73 to 1.15, n=1993, three trials, low certainty evidence) and loss to follow-up (RR 1.13, 95% CI 0.68 to 1.91, n=1993. three trials, low certainty evidence). We found some evidence that costs may be reduced for the patient and health system when task-shifting is undertaken. The low certainty regarding the evidence implies a high likelihood that further research may find the effects of the intervention to be substantially different from our findings. If resource-constrained countries decide to shift ART dispensing and distribution from pharmacy to non-pharmacy personnel, this should be accompanied by robust monitoring and impact evaluation.


HER/ErbB receptor interactions and signaling patterns in human mammary epithelial cells.
Knowledge about signaling pathways is typically compiled based on data gathered using different cell lines. This approach implicitly assumes that the cell line dependence is not important. However, different cell lines do not always respond to a particular stimulus in the same way, and lack of coherent data collected from closely related cellular systems can be detrimental to the efforts to understand the regulation of biological processes. To address this issue, we created a clone library of human mammary epithelial (HME) cells that expresses different levels of HER2 and HER3 receptors in combination with endogenous EGFR/HER1. Using our clone library, we have quantified the receptor activation patterns and systematically tested the validity of the existing hypotheses about the interaction patterns between HER1-3 receptors. Our study identified HER2 as the dominant dimerization partner for both EGFR and HER3. Contrary to earlier suggestions, we find that lateral interactions with HER2 do not lead to strong transactivation between EGFR and HER3, i.e., EGFR activation and HER3 activation are only weakly linked in HME cells. We also find that observed weak transactivation is uni-directional where stimulation of EGFR leads to HER3 activation whereas HER3 stimulation does not activate the EGFR. Repeating our experiments at lower cell confluency established that cell confluency is not a major factor in the observed interaction patterns. We have also quantified the dependence of the kinetics of Erk and Akt activation on different HER receptors. We found that HER3 signaling makes the strongest contribution to Akt activation and that, stimulation of either EGFR or HER3 leads to significant Erk activation. Our study shows that clone cell libraries can be a powerful resource in systems biology research by making it possible to differentiate between various hypotheses in a consistent cellular background. Using our constructed clone library we profiled the cell signaling patterns to establish the role of HER2 in the crosstalk between EGFR and HER3 receptors in HME cells. Our results for HME cells show that the weak linkage between EGFR and HER3 pathways can lead to distinct downstream cellular signaling patterns in response to the ligands of these two receptors.


A genome-wide association study of the Protein C anticoagulant pathway.
The Protein C anticoagulant pathway regulates blood coagulation by preventing the inadequate formation of thrombi. It has two main plasma components: protein C and protein S. Individuals with protein C or protein S deficiency present a dramatically increased incidence of thromboembolic disorders. Here, we present the results of a genome-wide association study (GWAS) for protein C and protein S plasma levels in a set of extended pedigrees from the Genetic Analysis of Idiopathic Thrombophilia (GAIT) Project. A total number of 397 individuals from 21 families were typed for 307,984 SNPs using the Infinium® 317 k Beadchip (Illumina). Protein C and protein S (free, functional and total) plasma levels were determined with biochemical assays for all participants. Association with phenotypes was investigated through variance component analysis. After correcting for multiple testing, two SNPs for protein C plasma levels (rs867186 and rs8119351) and another two for free protein S plasma levels (rs1413885 and rs1570868) remained significant on a genome-wide level, located in and around the PROCR and the DNAJC6 genomic regions respectively. No SNPs were significantly associated with functional or total protein S plasma levels, although rs1413885 from DNAJC6 showed suggestive association with the functional protein S phenotype, possibly indicating that this locus plays an important role in protein S metabolism. Our results provide evidence that PROCR and DNAJC6 might play a role in protein C and free protein S plasma levels in the population studied, warranting further investigation on the role of these loci in the etiology of venous thromboembolism and other thrombotic diseases.


Selection of endurance capabilities and the trade-off between pressure and volume in the evolution of the human heart.
Chimpanzees and gorillas, when not inactive, engage primarily in short bursts of resistance physical activity (RPA), such as climbing and fighting, that creates pressure stress on the cardiovascular system. In contrast, to initially hunt and gather and later to farm, it is thought that preindustrial human survival was dependent on lifelong moderate-intensity endurance physical activity (EPA), which creates a cardiovascular volume stress. Although derived musculoskeletal and thermoregulatory adaptations for EPA in humans have been documented, it is unknown if selection acted similarly on the heart. To test this hypothesis, we compared left ventricular (LV) structure and function across semiwild sanctuary chimpanzees, gorillas, and a sample of humans exposed to markedly different physical activity patterns. We show the human LV possesses derived features that help augment cardiac output (CO) thereby enabling EPA. However, the human LV also demonstrates phenotypic plasticity and, hence, variability, across a wide range of habitual physical activity. We show that the human LV's propensity to remodel differentially in response to chronic pressure or volume stimuli associated with intense RPA and EPA as well as physical inactivity represents an evolutionary trade-off with potential implications for contemporary cardiovascular health. Specifically, the human LV trades off pressure adaptations for volume capabilities and converges on a chimpanzee-like phenotype in response to physical inactivity or sustained pressure loading. Consequently, the derived LV and lifelong low blood pressure (BP) appear to be partly sustained by regular moderate-intensity EPA whose decline in postindustrial societies likely contributes to the modern epidemic of hypertensive heart disease.


Predictive value of the immediate effect of first dose of tamsulosin on lower urinary tract symptoms improvement in benign prostatic hyperplasia patients.
The objective is to study the effect of tamsulosin within hours after the first dose and its prediction of the future improvement of LUTS. From May 2016 until August 2017, 340 patients aged over 40 years with benign prostatic hyperplasia (BPH)-related symptoms were prospectively enrolled; 0.4 mg tamsulosin for 3 months was given. The first visit was before beginning of tamsulosin; uroflowmetry (UFM), postvoid residual urine volume (PVR), international prostate symptom score (IPSS), and quality of life (QoL) were measured. The second visit was after 6 h from the administration of tamsulosin. UFM and PVR were measured. The third visit was after 1 month and the fourth visit was after 3 months, on which UFM, PVR, IPSS, and QoL were also measured. The mean patients' age was 63 ± 6.18 and the mean prostate volume was 52.23 ± 24.59 cc. The mean Qmax at 1st, 2nd, 3rd, and 4th visits was 10.28 ± 3.06 s, 14.58 ± 4.84 s, 14.46 ± 4.94 s, and 14.28 ± 5.07 s, respectively, P = 0.04. The mean voiding time at 1st, 2nd, 3rd, and 4th visits was 41.24 ± 27.18 s, 33.84 ± 18.14 s, 31.96 ± 22.02 s, and 30.14 ± 17.52 s, respectively, P = 0.03. The mean PVR at 1st, 2nd, 3rd, and 4th visits was 46.40 ± 22.14 ml, 27.76 ± 26.10 ml, 25.16 ± 28.36 ml, and 25.58 ± 28.10 ml, respectively, P = 0.001. The first dose of tamsulosin significantly increases Qmax and decreases voiding time and residual urine (RU); there was no statistical significant difference between 1st dose, 1 and 3 months in Qmax, voiding time, and RU. QOL and IPSS were significantly improved after 1 and 3 months, P < 0.001. The first dose of tamsulosin improves UFM and predicts the mid-term change in UFM as well as IPSS and QoL indices in the treatment of BPH-related LUTS.


Meta-analysis: interferon for the treatment of chronic hepatitis C in dialysis patients.
The efficacy of interferon monotherapy in dialysis patients with chronic hepatitis C remains unclear, although a number of small clinical trials have been published addressing this issue. We evaluated the efficacy and safety of initial interferon monotherapy in dialysis patients with chronic hepatitis C by performing a systematic review of the literature with a meta-analysis of clinical trials. The primary outcome was sustained virological response (as a measure of efficacy); the secondary outcome was drop-out rate (as a measure of tolerability). We used the random effects model of Der Simonian and Laird, with heterogeneity and sensitivity analyses. We have identified 14 clinical trials (269 unique patients); two were controlled studies. The mean overall estimate for sustained virological response (SVR) and drop-out rate was 37%[95% confidence interval (CI) 28-48] and 17% (95% CI 10-28), respectively. The most frequent side-effects requiring interruption of treatment were flu-like symptoms (17%), neurological (21%) and gastrointestinal (18%). The overall weighted estimate for SVR in patients with hepatitis C virus genotype 1 was 30.6% (95% CI 20.9-48). In the sub-group of clinical trials (n = 5) with standard interferon administration (3 million units [MUI] thrice weekly, subcutaneous route, 24-week treatment), the overall mean estimate of SVR was 39% (95% CI 25-56). The studies were heterogeneous with regard to SVR and drop-out rate. Tolerance to initial interferon monotherapy was lower in dialysis than nonuremic patients with chronic hepatitis C. However, more than one-third of haemodialysis patients with chronic hepatitis C have been successfully treated with interferon. Longer duration of interferon monotherapy does not appear to have a beneficial effect on the response rate. Further studies are warranted to define the optimal anti-viral regimen for chronic hepatitis C in dialysis population.


Stereochemical studies of the monocyclic agouti-related protein (103-122) Arg-Phe-Phe residues: conversion of a melanocortin-4 receptor antagonist into an agonist and results in the discovery of a potent and selective melanocortin-1 agonist.
The agouti-related protein (AGRP) is an endogenous antagonist of the centrally expressed melanocortin receptors. The melanocortin-4 receptor (MC4R) is involved in energy homeostasis, food intake, sexual function, and obesity. The endogenous hAGRP protein is 132 amino acids in length, possesses five disulfide bridges at the C-terminus of the molecule, and is expressed in the hypothalamus of the brain. We have previously reported that a monocyclic hAGRP(103-122) peptide is an antagonist at the melanocortin receptors expressed in the brain. Stereochemical inversion from the endogenous l- to d-isomers of single or multiple amino acid modifications in this monocyclic truncated hAGRP sequence resulted in molecules that are converted from melanocortin receptor antagonists into melanocortin receptor agonists. The Asp-Pro-Ala-Ala-Thr-Ala-Tyr-cyclo[Cys-Arg-DPhe-DPhe-Asn-Ala-Phe-Cys]-Tyr-Ala-Arg-Lys-Leu peptide resulted in a 60 nM melanocortin-1 receptor agonist that is 100-fold selective versus the mMC4R, 1000-fold selective versus the mMC3R, and ca. 180-fold selective versus the mMC5R. In attempts to identify putative ligand-receptor interactions that may be participating in the agonist induced stimulation of the MC4R, selected ligands were docked into a homology molecular model of the mMC4R. These modeling studies have putatively identified hAGRP ligand DArg111-mMC4RAsn115 (TM3) and the hAGRP DPhe113-mMC4RPhe176 (TM4) interactions as important for agonist activity.


A multidimensional approach to measure poverty in rural Bangladesh.
Poverty is increasingly being understood as a multidimensional phenomenon. Other than income-consumption, which has been extensively studied in the past, health, education, shelter, and social involvement are among the most important dimensions of poverty. The present study attempts to develop a simple tool to measure poverty in its multidimensionality where it views poverty as an inadequate fulfillment of basic needs, such as food, clothing, shelter, health, education, and social involvement. The scale score ranges between 72 and 24 and is constructed in such a way that the score increases with increasing level of poverty. Using various techniques, the study evaluates the poverty-measurement tool and provides evidence for its reliability and validity by administering it in various areas of rural Bangladesh. The reliability coefficients, such as test-retest coefficient (0.85) and Cronbach's alpha (0.80) of the tool, were satisfactorily high. Based on the socioeconomic status defined by the participatory rural appraisal (PRA) exercise, the level of poverty identified by the scale was 33% in Chakaria, 26% in Matlab, and 32% in other rural areas of the country. The validity of these results was tested against some traditional methods of identifying the poor, and the association of the scores with that of the traditional indicators, such as ownership of land and occupation, asset index (r=0.72), and the wealth ranking obtained from the PRA exercise, was consistent. A statistically significant inverse relationship of the poverty scores with the socioeconomic status was observed in all cases. The scale also allowed the absolute level of poverty to be measured, and in the present study, the highest percentage of absolute poor was found in terms of health (44.2% in Chakaria, 36.4% in Matlab, and 39.1% in other rural areas), followed by social exclusion (35.7% in Chakaria, 28.5% in Matlab, and 22.3% in other rural areas), clothing (6.2% in Chakaria, 8.3% in Matlab, and 20% in other rural areas), education (14.7% in Chakaria, 8% in Matlab, and 16.8% in other rural areas), food (7.8% in Chakaria, 2.9% in Matlab and 3% in other rural areas), and shelter (0.8% in Chakaria, 1.4% in Matlab, and 3.7% in other rural areas). This instrument will also prove itself invaluable in assessing the individual effects of poverty-alleviation programmes or policies on all these different dimensions.


Glen W. Hartman Lecture. Science, creativity, and serendipity.
In this Roentgen centennial year, honoring the epochal discovery of the X-ray in 1895 [2], it is particularly appropriate to reflect on the circumstances of creativity that lead to important discoveries in science. In radiology as in general medicine and surgery, histories of the development of the discipline have generally presented a chronicle of sequential events without specific consideration of the underlying creative impulse [3-7]. I wish to consider features of creativity that may be relevant to us, that may enlighten us, and that may allow us to incorporate them individually into our view of the world and our appreciation of the grandeur of such contributions.


Loss-of-function mutations in the keratin 5 gene lead to Dowling-Degos disease.
Dowling-Degos disease (DDD) is an autosomal dominant genodermatosis characterized by progressive and disfiguring reticulate hyperpigmentation of the flexures. We performed a genomewide linkage analysis of two German families and mapped DDD to chromosome 12q, with a total LOD score of 4.42 ( theta =0.0) for marker D12S368. This region includes the keratin gene cluster, which we screened for mutations. We identified loss-of-function mutations in the keratin 5 gene (KRT5) in all affected family members and in six unrelated patients with DDD. These represent the first identified mutations that lead to haploinsufficiency in a keratin gene. The identification of loss-of-function mutations, along with the results from additional functional studies, suggest a crucial role for keratins in the organization of cell adhesion, melanosome uptake, organelle transport, and nuclear anchorage.


Transcriptome analysis of amoeboid and ramified microglia isolated from the corpus callosum of rat brain.
Microglia, the resident immune cells of the central nervous system (CNS), have two distinct phenotypes in the developing brain: amoeboid form, known to be amoeboid microglial cells (AMC) and ramified form, known to be ramified microglial cells (RMC). The AMC are characterized by being proliferative, phagocytic and migratory whereas the RMC are quiescent and exhibit a slow turnover rate. The AMC transform into RMC with advancing age, and this transformation is indicative of the gradual shift in the microglial functions. Both AMC and RMC respond to CNS inflammation, and they become hypertrophic when activated by trauma, infection or neurodegenerative stimuli. The molecular mechanisms and functional significance of morphological transformation of microglia during normal development and in disease conditions is not clear. It is hypothesized that AMC and RMC are functionally regulated by a specific set of genes encoding various signaling molecules and transcription factors. To address this, we carried out cDNA microarray analysis using lectin-labeled AMC and RMC isolated from frozen tissue sections of the corpus callosum of 5-day and 4-week old rat brain respectively, by laser capture microdissection. The global gene expression profiles of both microglial phenotypes were compared and the differentially expressed genes in AMC and RMC were clustered based on their functional annotations. This genome wide comparative analysis identified genes that are specific to AMC and RMC. The novel and specific molecules identified from the trancriptome explains the quiescent state functioning of microglia in its two distinct morphological states.


Marker rescue of the host range restriction defects of modified vaccinia virus Ankara.
The severely attenuated and host range (HR) restricted modified vaccinia virus Ankara (MVA) was derived by >500 passages in chick embryo fibroblasts, during which multiple deletions and mutations occurred. To determine the basis of the HR defect, we prepared cosmids from the parental vaccinia virus Ankara genome and transfected them into nonpermissive monkey BS-C-1 cells that had been infected with MVA. Recombinant viruses that formed macroscopic plaques were detected after transfections with DNA segments that mapped near the left end of the viral genome. Plaque-forming viruses, generated by transfections with four individual cosmids and one pair of minimally overlapping cosmids, were purified, and their HRs were evaluated in BS-C-1 cells, rabbit RK-13 cells, and human HeLa, MRC-5, and A549 cells. The acquisition of the K1L and SPI-1 HR genes and the repair of large deletions were determined by polymerase chain reaction or pulse-field gel electrophoresis of NotI restriction enzyme digests of genomic DNA. The following results indicated the presence of previously unrecognized vaccinia virus HR genes: (1) the major mutations that restrict HR are within the left end of the MVA genome because the phenotypes of some recombinants approached that of the parental virus, (2) acquisition of the K1L gene correlated with the ability of recombinant viruses to propagate in RK-13 cells but did not enhance replication in human or monkey cell lines, (3) acquisition of the SPI-1 gene correlated with virus propagation in A549 cells but not with the extent of virus spread in monkey or other human cell lines, (4) there are at least two impaired HR genes because rescue occurred with nonoverlapping or minimally overlapping cosmids and recombinant viruses with intermediate HRs were isolated, and (5) at least one of the new HR genes did not map within any of the major deletions because the size of the left terminal NotI fragment was not appreciably altered in some recombinant viruses.


Social structure mediates environmental effects on group size in an obligate cooperative breeder, Suricata suricatta.
Population dynamics in group-living species can be strongly affected both by features of sociality per se and by resultant population structure. To develop a mechanistic understanding of population dynamics in highly social species we need to investigate how processes within groups, processes linking groups, and external drivers act and interact to produce observed patterns. We model social group dynamics in cooperatively breeding meerkats, Suricata suricatta, paying attention to local demographic as well as dispersal processes. We use generalized additive models to describe the influence of group size, population density, and environmental conditions on demographic rates for each sex and stage, and we combine these models into predictive and individual-based simulation models of group dynamics. Short-term predictions of expected group size and simulated group trajectories over the longer term agree well with observations. Group dynamics are characterized by slow increases during the breeding season and relatively sharp declines during the pre-breeding season, particularly after dry years. We examine the demographic mechanisms responsible for environmental dependence. While individuals appear more prone to emigrate after dry years, seasons of low rainfall also cause reductions in reproductive output that produce adult-biased age distributions in the following dispersal season. Adult subordinates are much more likely to disperse or be evicted than immature individuals, and demographic structure thus contributes to crashes in group size. Our results demonstrate the role of social structure in characterizing a population's response to environmental variation. We discuss the implications of our findings for the population dynamics of cooperative breeders and population dynamics generally.


A large outbreak of botulism: the hazardous baked potato.
In April 1994, the largest outbreak of botulism in the United States since 1978 occurred in El Paso, Texas. Thirty persons were affected; 4 required mechanical ventilation. All ate food from a Greek restaurant. The attack rate among persons who ate a potato-based dip was 86% (19/22) compared with 6% (11/176) among persons who did not eat the dip (relative risk [RR] = 13.8; 95% confidence interval [CI], 7.6-25.1). The attack rate among persons who ate an eggplant-based dip was 67% (6/9) compared with 13% (241189) among persons who did not (RR = 5.2; 95% CI, 2.9-9.5). Botulism toxin type A was detected from patients and in both dips. Toxin formation resulted from holding aluminum foil-wrapped baked potatoes at room temperature, apparently for several days, before they were used in the dips. Consumers should be informed of the potential hazards caused by holding foil-wrapped potatoes at ambient temperatures after cooking.


Signaling mechanisms of LOV domains: new insights from molecular dynamics studies.
Phototropins are one of several classes of photoreceptors used by plants and algae to respond to light. These proteins contain flavin-binding LOV (Light-Oxygen-Voltage) domains that form covalent cysteine-flavin adducts upon exposure to blue light, leading to the enhancement of phototropin kinase activity. Several lines of evidence suggest that adduct formation in the phototropin LOV2 domains leads to the dissociation of an alpha helix (Jα) from these domains as part of the light-induced activation process. However, crystal structures of LOV domains both in the presence and absence of the Jα helix show very few differences between dark and illuminated states, and thus the precise mechanism through which adduct formation triggers helical dissociation remains poorly understood. Using Avena sativa phototropin 1 LOV2 as a model system, we have studied the interactions of the LOV domain core with the Jα helix through a series of equilibrium molecular dynamics simulations. Here we show that conformational transitions of a conserved glutamine residue in the flavin binding pocket are coupled to altered dynamics of the Jα helix both through a shift in dynamics of the main β-sheet of the LOV domain core and through a secondary pathway involving the N-terminal A'α helix.


Alternative variants of human HYDIN are novel cancer-associated antigens recognized by adaptive immunity.
A mutation in the hydin gene has been recently described as one possible mechanism leading to lethal congenital hydrocephalus in mice, and a similar defect is proposed to be involved in an autosomal recessive form of hydrocephalus in human. Here, we report for the first time on the cancer association and immunogenicity of two HYDIN variants in humans. One is a previously described sequence derived from the chromosome 1 gene copy, that is, KIAA1864. The second is encoded by a novel alternative transcript originating from the chromosome 16, which we identified by immunoscreening of a testis-derived cDNA expression library with sera of patients with colorectal cancer, and called MO-TES391. Both variants are targeted by immunoglobulin G antibodies in a significant subset of cancer patients but only rarely in healthy donors. Moreover, we identify HLA-A*0201-restricted sequences derived from MO-TES391 and KIAA1864, which are specifically recognized by human cytotoxic CD8(+) T cells. Taken together, these results suggest frequent and coordinated adaptive immune responses against HYDIN variants in patients with cancer and propose HYDIN as a novel cancer-associated antigen.


T-helper-1-cell cytokines drive cancer into senescence.
Cancer control by adaptive immunity involves a number of defined death and clearance mechanisms. However, efficient inhibition of exponential cancer growth by T cells and interferon-γ (IFN-γ) requires additional undefined mechanisms that arrest cancer cell proliferation. Here we show that the combined action of the T-helper-1-cell cytokines IFN-γ and tumour necrosis factor (TNF) directly induces permanent growth arrest in cancers. To safely separate senescence induced by tumour immunity from oncogene-induced senescence, we used a mouse model in which the Simian virus 40 large T antigen (Tag) expressed under the control of the rat insulin promoter creates tumours by attenuating p53- and Rb-mediated cell cycle control. When combined, IFN-γ and TNF drive Tag-expressing cancers into senescence by inducing permanent growth arrest in G1/G0, activation of p16INK4a (also known as CDKN2A), and downstream Rb hypophosphorylation at serine 795. This cytokine-induced senescence strictly requires STAT1 and TNFR1 (also known as TNFRSF1A) signalling in addition to p16INK4a. In vivo, Tag-specific T-helper 1 cells permanently arrest Tag-expressing cancers by inducing IFN-γ- and TNFR1-dependent senescence. Conversely, Tnfr1(-/-)Tag-expressing cancers resist cytokine-induced senescence and grow aggressively, even in TNFR1-expressing hosts. Finally, as IFN-γ and TNF induce senescence in numerous murine and human cancers, this may be a general mechanism for arresting cancer progression.


Regulatory domain selectivity in the cell-type specific PKN-dependence of cell migration.
The mammalian protein kinase N (PKN) family of Serine/Threonine kinases comprises three isoforms, which are targets for Rho family GTPases. Small GTPases are major regulators of the cellular cytoskeleton, generating interest in the role(s) of specific PKN isoforms in processes such as cell migration and invasion. It has been reported that PKN3 is required for prostate tumour cell invasion but not PKN1 or 2. Here we employ a cell model, the 5637 bladder tumour cell line where PKN2 is relatively highly expressed, to assess the potential redundancy of these isoforms in migratory responses. It is established that PKN2 has a critical role in the migration and invasion of these cells. Furthermore, using a PKN wild-type and chimera rescue strategy, it is shown that PKN isoforms are not simply redundant in supporting migration, but appear to be linked through isoform specific regulatory domain properties to selective upstream signals. It is concluded that intervention in PKNs may need to be directed at multiple isoforms to be effective in different cell types.


Exercise training normalizes an increased neuronal excitability of NTS-projecting neurons of the hypothalamic paraventricular nucleus in hypertensive rats.
Elevated sympathetic outflow and altered autonomic reflexes, including impaired baroreflex function, are common findings observed in hypertensive disorders. Although a growing body of evidence supports a contribution of preautonomic neurons in the hypothalamic paraventricular nucleus (PVN) to altered autonomic control during hypertension, the precise underlying mechanisms remain unknown. Here, we aimed to determine whether the intrinsic excitability and repetitive firing properties of preautonomic PVN neurons that innervate the nucleus tractus solitarii (PVN-NTS neurons) were altered in spontaneously hypertensive rats (SHR). Moreover, given that exercise training is known to improve and/or correct autonomic deficits in hypertensive conditions, we evaluated whether exercise is an efficient behavioral approach to correct altered neuronal excitability in hypertensive rats. Patch-clamp recordings were obtained from retrogradely labeled PVN-NTS neurons in hypothalamic slices obtained from sedentary (S) and trained (T) Wistar-Kyoto (WKY) and SHR rats. Our results indicate an increased excitability of PVN-NTS neurons in SHR-S rats, reflected by an enhanced input-output function in response to depolarizing stimuli, a hyperpolarizing shift in Na(+) spike threshold, and smaller hyperpolarizing afterpotentials. Importantly, we found exercise training in SHR rats to restore all these parameters back to those levels observed in WKY-S rats. In several cases, exercise evoked opposing effects in WKY-S rats compared with SHR-S rats, suggesting that exercise effects on PVN-NTS neurons are state dependent. Taken together, our results suggest that elevated preautonomic PVN-NTS neuronal excitability may contribute to altered autonomic control in SHR rats and that exercise training efficiently corrects these abnormalities.


Cloning of a palmitoyl-acyl carrier protein thioesterase from oil palm.
A palmitoyl-acyl carrier protein (ACP) thioesterase cDNA clone was isolated from an oil palm cDNA library. The cDNA was expressed in Escherichia coli as a glutathione S-transferase fusion protein and a crude bacterial extract was assayed for acyl-CoA-hydrolysing activity. The recombinant enzyme was able to hydrolyse medium- and long-chain acyl-CoAs. Northern-blot analysis showed a high level of gene expression in leaf, flower and 15-, 17- and 18-week mesocarp tissues. Low-level gene expression was detected in germinated seedlings and 8- and 12-week mesocarp tissues, but no transcript was detected in any kernel tissues. Southern-blot analysis indicated the presence of a single gene and we have also isolated a genomic clone using the cDNA as a probe. Two genomic fragments were subcloned and a 7 kb contiguous stretch of the oil palm genome was sequenced. Comparison of this sequence with the cDNA sequence identified a putative 93 amino acid transit peptide, most of which is missing from the cDNA. The coding region of the gene consisted of seven exons and six introns.


Activation of a bacterial virulence protein by the GTPase RhoA.
The Rho family of guanosine triphosphatases (GTPases) are essential eukaryotic signaling molecules that regulate cellular physiology. Virulence factors from various pathogens alter the signaling of GTPases by acting as GTPase activating factors, guanine nucleotide exchange factors, or direct covalent modifiers; however, bacterial virulence factors that sense rather than alter the signaling states of Rho GTPases have not been previously described. Here, we report that the translocated Salmonellae virulence factor SseJ binds to the guanosine triphosphate-bound form of RhoA. This interaction stimulates the lipase activity of SseJ, which results in the esterification of cholesterol in the host cell membrane. Our results suggest that the activation of molecules downstream of GTPases is not exclusive to eukaryotic proteins, and that a bacterial protein has evolved to recognize the activation state of RhoA, which regulates its enzymatic activity as part of the host-pathogen interaction.


Glycerol-Free Cryopreservation of Red Blood Cells Enabled by Ice-Recrystallization-Inhibiting Polymers.
Cryopreservation is fundamental in prolonging the viabilities of cells and tissues of clinical and biotechnological relevance ex vivo. Furthermore, there is an increasing need to address storage at more easily accessible temperatures in the developing world because of limited resources. Here, the cryopreservation of erythrocytes (red blood cells) with storage at -20 °C using hydroxyethyl starch (HES) and the ice recrystallization inhibitor poly(vinyl alcohol) (PVA), which is a biomimetic of naturally occurring antifreeze (glyco)proteins (AF(G)Ps), is described. This strategy eliminates the need for high concentrations of membrane penetrating solvents such as glycerol or dimethyl sulfoxide (DMSO). The addition of only 0.1-0.5 wt % PVA to the polymeric cryoprotectant, HES, significantly enhances cell recovery under conditions that promote damage due to ice recrystallization. The comparative ease with which the addition and removal of both HES and PVA can be attained is an additional attractive quality. Coupled with the benefits attained by the ice recrystallization inhibition activity of PVA, this methodology therefore offers a strategy that could aid the storage and distribution of biological materials.


Cultural competence in medical education: A questionnaire study of Danish medical teachers' perceptions of and preparedness to teach cultural competence.
The cultural competence training of healthcare professionals is a key element in ensuring the quality of both the access and delivery of healthcare to increasingly ethnically diverse populations. The aim of this study is to investigate Danish medical teachers' opinions about cultural competence, their willingness to receive training and preparedness to teach cultural competence topics. The survey was sent to medical teachers, clinical teachers and external lecturers who teach in the medical programme at the University of Copenhagen. A total of 1400 medical teachers received the survey, and 199 responded. The response rate is 14%. Data were analysed through descriptive calculations, and answers to open-ended questions were coded using content analysis. Results showed that 82.4% of the informants agreed or strongly agreed that the medical education programme should include training on cultural issues, and 60.3% agreed or strongly agreed that students should be assessed on their cultural competence skills. Regarding preparedness to teach a diverse classroom, 88.4% felt somewhat or very prepared to engage and motivate all students. About 70% were interested in receiving training on cultural competence. Generally, there is interest in and acknowledgement of the importance of cultural competence in Danish medical education among teachers at the University of Copenhagen. This creates an opportunity to implement cultural competence in the medical curriculum, training of teachers and strengthening the diversity sensitivity of the organisation. However, support for this programme by management and the allocation of an appropriate level of resources is a prerequisite to the success of the programme.


Pollen derived low molecular compounds enhance the human allergen specific immune response in vivo.
Besides allergens, pollen release bioactive, low molecular weight compounds that modulate and stimulate allergic reactions. Clinical relevance of these substances has not been investigated to date. To elucidate the effect of a non-allergenic, low molecular weight factors from aqueous birch pollen extracts (Bet-APE < 3 kDa) on the human allergic immune response in vivo. Birch and grass pollen allergic individuals underwent skin prick testing with allergen alone, allergen plus Bet-APE < 3 kDa, or allergen plus pre-identified candidate substances from low molecular pollen fraction. Nasal allergen challenges were performed in non-atopic and pollen allergic individuals using a 3 day repeated threshold challenge battery. Subjects were either exposed to allergen alone or to allergen plus Bet-APE< 3 kDa. Local cytokine levels, nasal secretion weights, nasal congestion and symptom scores were determined. Skin prick test reactions to pollen elicited larger weals when allergens were tested together with the low molecular weight compounds from pollen. Similar results were obtained with candidate pollen-associated lipid mediators. In nasal lining fluids of allergic patients challenged with allergen plus Bet-APE < 3 kDa, IL-8 and IgE was significantly increased as compared to allergen-only challenged patients. These patients also produced increased amounts of total nasal secretion and reported more severe rhinorrhea than the allergen-only challenged group. Low molecular compounds from pollen enhance the allergen specific immune response in the skin and nose. They are therefore of potential clinical relevance in allergic patients.


Crossing the Communication Chasm: Challenges and Opportunities in Transitions of Care from the Hospital to the Primary Care Clinic.
Transitions of care from specialty and acute settings to primary care abound. Compared to the continuity in end-of-shift handoffs, care transitions involve provider communication between practices and facilities with their own cultures and bureaucracies. Using the transition from acute care to outpatient primary care for stroke/transient ischemic attack (TIA) patients as a case study, this qualitative research explored communication practices and institutional arrangements among clinical providers responsible for longitudinal management of hypertension. In this study, researchers investigated the barriers and facilitators of effective communication between acute stroke/TIA inpatient and primary care providers at a Veterans Affairs Medical Center. A multidisciplinary team conducted consensus-based coding and thematic analysis of semistructured interviews with 21 clinical providers (9 with primary responsibilities for inpatient care and 12 with primary responsibilities in outpatient, primary care). Thematic analysis of responses identified three factors that influenced communication between clinical providers: (1) consistent, concise but complete medication and treatment plans; (2) reliable, standardized discharge documentation; (3) use of multiple modes of communication. Participants identified cultural barriers, including challenges with rotating providers at a teaching hospital and local discharge practices. Ambiguity about who is being handed off to and time pressures in the acute setting may lead inpatient providers to give lower priority to discharge communication, leaving outpatient providers with low-quality information. While electronic templates have standardized key components of discharge documentation, improvement opportunities remain. Increased awareness of the challenges and opportunities on each side of the care transfer could foster communication practices that systematically account for the information needs of inpatient and outpatient providers.


The Rhodococcus opacus TadD protein mediates triacylglycerol metabolism by regulating intracellular NAD(P)H pools.
The Gram-positive actinomycete Rhodococcus opacus is widely studied for its innate ability to store large amounts of carbon in the form of triacylglycerol (TAG). Several groups have demonstrated that R. opacus PD630 is capable of storing anywhere from 50 to 76% of its cell dry weight as TAG. While numerous studies have focused on phenomenological aspects of this process, few have sought to identify the underlying molecular and biochemical mechanisms responsible for the biosynthesis and storage of this molecule. Herein we further our previous efforts to illuminate the black box that is lipid metabolism in actinomycetes using a genetic approach. Utilizing a simple, colorimetric genetic screen, we have identified a gene, referred to herein as tadD (triacylglycerol accumulation deficient), which is critical for TAG biosynthesis in R. opacus PD630. Furthermore, we demonstrate that the purified protein product of this gene is capable of oxidizing glyceraldehyde-3-phosphate, while simultaneously reducing NAD(P)+ to NAD(P)H. Supporting this biochemical data, we observed that the ratio of NAD(P)H to NAD(P)+ is elevated in wildtype cultures grown under lipid production conditions as compared to cultures grown under vegetative growth conditions, while the mutant strain demonstrated no change irrespective of growth conditions. Finally, we demonstrate that over-expressing a putative phosphorylative glyceraldehyde-3-phosphate dehydrogenase leads to decreased TAG production during growth on TAG accumulation conditions. Taken together, the data support the identification of a key metabolic branch point separating vegetative growth and lipid accumulation lifestyles in Rhodococcus.


Epidemiology of Alzheimer's disease and other forms of dementia in China, 1990-2010: a systematic review and analysis.
China is increasingly facing the challenge of control of the growing burden of non-communicable diseases. We assessed the epidemiology of Alzheimer's disease and other forms of dementia in China between 1990, and 2010, to improve estimates of the burden of disease, analyse time trends, and inform health policy decisions relevant to China's rapidly ageing population. In our systematic review we searched for reports of Alzheimer's disease or dementia in China, published in Chinese and English between 1990 and 2010. We searched China National Knowledge Infrastructure, Wanfang, and PubMed databases. Two investigators independently assessed case definitions of Alzheimer's disease and dementia: we excluded studies that did not use internationally accepted case definitions. We also excluded reviews and viewpoints, studies with no numerical estimates, and studies not done in mainland China. We used Poisson regression and UN demographic data to estimate the prevalence (in nine age groups), incidence, and standardised mortality ratio of dementia and its subtypes in China in 1990, 2000, and 2010. Our search returned 12,642 reports, of which 89 met the inclusion criteria (75 assessed prevalence, 13 incidence, and nine mortality). In total, the included studies had 340,247 participants, in which 6357 cases of Alzheimer's disease were recorded. 254,367 people were assessed for other forms of dementia, of whom 3543 had vascular dementia, frontotemporal dementia, or Lewy body dementia. In 1990 the prevalence of all forms of dementia was 1·8% (95% CI 0·0-44·4) at 65-69 years, and 42·1% (0·0-88·9) at age 95-99 years. In 2010 prevalence was 2·6% (0·0-28·2) at age 65-69 years and 60·5% (39·7-81·3) at age 95-99 years. The number of people with dementia in China was 3·68 million (95% CI 2·22-5·14) in 1990, 5·62 million (4·42-6·82) in 2000, and 9·19 million (5·92-12·48) in 2010. In the same period, the number of people with Alzheimer's disease was 1·93 million (1·15-2·71) in 1990, 3·71 million (2·84-4·58) people in 2000, and 5·69 million (3·85-7·53) in 2010. The incidence of dementia was 9·87 cases per 1000 person-years, that of Alzheimer's disease was 6·25 cases per 1000 person-years, that of vascular dementia was 2·42 cases per 1000 person-years, and that of other rare forms of dementia was 0·46 cases per 1000 person-years. We retrieved mortality data for 1032 people with dementia and 20,157 healthy controls, who were followed up for 3-7 years. The median standardised mortality ratio was 1·94:1 (IQR 1·74-2·45). Our analysis suggests that previous estimates of dementia burden, based on smaller datasets, might have underestimated the burden of dementia in China. The burden of dementia seems to be increasing faster than is generally assumed by the international health community. Rapid and effective government responses are needed to tackle dementia in low-income and middle-income countries. Nossal Institute of Global Health (University of Melbourne, Australia), the National 12th Five-Year Major Projects of China, National Health and Medical Research Council Australia-China Exchange Fellowship, Importation and Development of High-Calibre Talents Project of Beijing Municipal Institutions, and the Bill & Melinda Gates Foundation.


Apparent Fibre Density: a novel measure for the analysis of diffusion-weighted magnetic resonance images.
This article proposes a new measure called Apparent Fibre Density (AFD) for the analysis of high angular resolution diffusion-weighted images using higher-order information provided by fibre orientation distributions (FODs) computed using spherical deconvolution. AFD has the potential to provide specific information regarding differences between populations by identifying not only the location, but also the orientations along which differences exist. In this work, analytical and numerical Monte-Carlo simulations are used to support the use of the FOD amplitude as a quantitative measure (i.e. AFD) for population and longitudinal analysis. To perform robust voxel-based analysis of AFD, we present and evaluate a novel method to modulate the FOD to account for changes in fibre bundle cross-sectional area that occur during spatial normalisation. We then describe a novel approach for statistical analysis of AFD that uses cluster-based inference of differences extended throughout space and orientation. Finally, we demonstrate the capability of the proposed method by performing voxel-based AFD comparisons between a group of Motor Neurone Disease patients and healthy control subjects. A significant decrease in AFD was detected along voxels and orientations corresponding to both the corticospinal tract and corpus callosal fibres that connect the primary motor cortices. In addition to corroborating previous findings in MND, this study demonstrates the clear advantage of using this type of analysis by identifying differences along single fibre bundles in regions containing multiple fibre populations.


The maternal social environment shapes offspring growth, physiology, and behavioural phenotype in guinea pigs.
Prenatal conditions influence offspring development in many species. In mammals, the effects of social density have traditionally been considered a detrimental form of maternal stress. Now their potential adaptive significance is receiving greater attention.Sex-specific effects of maternal social instability on offspring in guinea pigs (Cavia aperea f. porcellus) have been interpreted as adaptations to high social densities, while the effects of low social density are unknown. Hence, we compared morphological, behavioural and physiological development between offspring born to mothers housed either individually or in groups during the second half of pregnancy. Females housed individually and females housed in groups gave birth to litters of similar size and sex-ratios, and there were no differences in birth weight. Sons of individually-housed mothers grew faster than their sisters, whereas daughters ofgroup-housed females grew faster than their brothers, primarily due to an effect on growth of daughters. There were few effects on offspring behaviour. Baseline cortisol levels in saliva of pups on day 1 and day 7 were not affected, but we saw a blunted cortisol response to social separation on day 7 in sons of individually-housed females and daughters of group-housed females. The effects were consistent across two replicate experiments. The observed effects only partially support the adaptive hypothesis. Increased growth of daughters may be adaptive under high densities due to increasedfemale competition, but it is unclear why growth of sons is not increased under low social densities when males face less competition from older, dominant males. The differences in growth may be causally linked to sex-specific effects on cortisol response, although individual cortisol response and growth were not correlated, and various other mechanisms are possible. The observed sex-specific effects on early development are intriguing, yet the potential adaptive benefits and physiological mechanisms require further study.


Investigation of factors associated with manual wheelchair mobility in persons with spinal cord injury.
To quantify wheelchair mobility of persons with a spinal cord injury (SCI), and to assess the relationship between wheelchair mobility and demographics, type of manual wheelchair, and participation. Cross-sectional study. Six Model Spinal Cord Injury Systems. People (N=132) with SCI who use a manual wheelchair as their primary means of mobility. Not applicable. Wheelchair-related mobility characteristics measured by a data-logging device, and community participation measured by the short form of the Craig Handicap Assessment Recording Technique (CHART). Age was found to be significantly (r=-.225, P<.01) related to average speed traveled per day. Whites were found to travel significantly further (P<.01) and accumulate more minutes per day (P<.01) compared with minorities. Participants who were employed traveled significantly further (P<.01), faster (P<.01), and for more minutes per day (P<.01) compared with those who were not employed. A moderate relationship (r=.245-.390) was found between wheelchair mobility data and CHART total score. Results suggest a need for future investigation of the factors that influence wheelchair mobility and community participation of persons with SCI. Findings indicate the efficacy of a quantitative method to track wheelchair mobility in community settings, which could serve as a way of identifying community participation for persons with SCI and possibly uncovering additional aspects of participation.


Screening of Asteraceae (Compositae) plant extracts for larvicidal activity against Aedes fluviatilis (Diptera:Culicidae).
Ethanol extracts of 83 plants species belonging to the Asteraceae (Compositae) family, collected in the State of Minas Gerais, Brazil, were tested for larvicidal activity against the mosquito Aedes fluviatilis--Diptera: Culicidae). The extract from Tagetes minuta was the most active with a LC90 of 1.5 mg/l and LC50 of 1.0 mg/l. This plant has been the object of several studies by other groups and its active components have already been identified as thiophene derivatives, a class of compounds present in many Asteraceae species. The extract of Eclipta paniculata was also significantly active, with a LC90 of 17.2 mg/l and LC50 of 3.3 mg/l and no previous studies on its larvicidal activity or chemical composition could be found in the literature. Extracts of Achryrocline satureoides, Gnaphalium spicatum, Senecio brasiliensis, Trixis vauthieri, Tagetes patula and Vernonia ammophila were less active, killing more than 50% of the larvae only at the higher dose tested (100 mg/l).


Efficacy of statin monotherapy or in combination with coenzyme a capsule in patients with metabolic syndrome and mixed dyslipidemia.
Patients with metabolic syndrome are at increased risk for cardiovascular disease. Combination lipid-lowering therapy is often needed in patients with metabolic syndrome and mixed dyslipidemia. The aim of this study was to compare the effect of statin combined with a new hypolipidemic agent, coenzyme A (CoA) with moderate-dose statin monotherapy in subjects with metabolic syndrome and mixed dyslipidemia by evaluating data from a subgroup of patients with metabolic syndrome and mixed dyslipidemia from a previously conducted randomized study. In the present post hoc analysis, 212 patients were included, receiving statin monotherapy (n = 94) or statin combined with CoA 400 U/day (n = 118) for 8 weeks. The lipoprotein profile was determined at baseline and week 8 visits. Attainment of low-density lipoprotein-cholesterol (LDL-C) < 100 mg/dL, non-high-density lipoprotein-cholesterol (HDL-C) < 130 mg/dL, and the combined goal of these two parameters was also evaluated. The mean percent change was more prominent with CoA plus statin compared with placebo plus statin in triglyceride (TG) (-32.5% vs. -8.7%, respectively; P = 0.0002), total cholesterol (-9.6% vs. -3.6%, P = 0.013), LDL-C (-7.5% vs. 2.1%, P = 0.033), and non-HDL-C (-14.3% vs. -6.4%, P = 0.011). Treatment with CoA plus statin resulted in larger percentages of participants attaining lipid goals for LDL-C (70.3% vs. 56.4%, P = 0.044), non-HDL-C (60.2% vs. 45.7%, P = 0.039), and the combined goal of LDL-C and non-HDL-C (57.6% vs. 42.6%, P = 0.038) than statin monotherapy. These results demonstrate that CoA plus statin therapy was more effective in improving lipoprotein parameters than statin alone in patients with metabolic syndrome and mixed hyperlipidemia.


Interaction Between Air Pollutants and Pollen Grains: The Role on the Rising Trend in Allergy.
Asthma and allergic diseases cases have risen in recent decades. Plant pollen is considered as the main aeroallergen causing allergic reactions. According to available data, urban residents experience more respiratory allergies than rural residents mainly due to the interaction between chemical air pollutants and pollen grains. This interaction can occur through several mechanisms; chemical pollutants might facilitate pollen allergen release, act as adjuvants to stimulate IgE-mediated responses, modify allergenic potential, and enhance the expression of some allergens in pollen grains. This review focuses on the most recent theories explaining how air pollutants can interact with pollen grains and allergens.


Pristimerin, a triterpenoid, inhibits tumor angiogenesis by targeting VEGFR2 activation.
Pristimerin is a triterpenoid isolated from Celastrus and Maytenus spp. that has been shown to possess a variety of biological activities, including anti-cancer activity. However, little is known about pristimerin's effects on tumor angiogenesis. In this study, we examined the function and the mechanism of this compound in tumor angiogenesis using multiple angiogenesis assays. We found that pristimerin significantly reduced both the volume and weight of solid tumors and decreased angiogenesis in a xenograft mouse tumor model in vivo. Pristimerin significantly inhibited the neovascularization of chicken chorioallantoic membrane (CAM) in vivo and abrogated vascular endothelial growth factor (VEGF)-induced microvessel sprouting in an ex vivo rat aortic ring assay. Furthermore, pristimerin inhibited the VEGF-induced proliferation, migration and capillary-like structure formation of human umbilical vascular endothelial cells (HUVECs) in a concentration-dependent manner. Mechanistic studies revealed that pristimerin suppressed the VEGF-induced phosphorylation of VEGF receptor 2 kinase (KDR/Flk-1) and the activity of AKT, ERK1/2, mTOR, and ribosomal protein S6 kinase. Taken together, our results provide evidence for the first time that pristimerin potently suppresses angiogenesis by targeting VEGFR2 activation. These results provide a novel mechanism of action for pristimerin which may be important in the treatment of cancer.


A global response to a global problem: the epidemic of overnutrition.
It is estimated that by 2020 two-thirds of the global burden of disease will be attributable to chronic noncommunicable diseases, most of them strongly associated with diet. The nutrition transition towards refined foods, foods of animal origin, and increased fats plays a major role in the current global epidemics of obesity, diabetes and cardiovascular diseases, among other noncommunicable conditions. Sedentary lifestyles and the use of tobacco are also significant risk factors. The epidemics cannot be ended simply by encouraging people to reduce their risk factors and adopt healthier lifestyles, although such encouragement is undoubtedly beneficial if the targeted people can respond. Unfortunately, increasingly obesogenic environments, reinforced by many of the cultural changes associated with globalization, make even the adoption of healthy lifestyles, especially by children and adolescents, more and more difficult. The present paper examines some possible mechanisms for, and WHO's role in, the development of a coordinated global strategy on diet, physical activity and health. The situation presents many countries with unmanageable costs. At the same time there are often continuing problems of undernutrition. A concerted multisectoral approach, involving the use of policy, education and trade mechanisms, is necessary to address these matters.


Eliciting willingness to pay: comparing closed-ended with open-ended and payment scale formats.
Willingness to pay (WTP) is increasingly being used as a measure of valuation in health technology assessment. A variety of formats for eliciting values are available, although the relative virtues of each remain the subject of methodological controversy. This article compares valuation results obtained using a WTP survey instrument in a closed-ended format with those obtained from instruments using open-ended and payment scale formats. Samples of subjects were drawn from a general population, and all were asked to value the same intervention--alternative methods of screening for colorectal cancer. It was discovered that, whereas the open-ended and payment scale formats produced broadly similar valuations, the closed-ended format produced significantly higher WTP valuations and different justifications for those valuations. It is hypothesized that anchoring and yea-saying effects explain these differences and that the closed-ended format triggers a different response mode in subjects.


A new method to deliver supraclavicular radiation in breast radiotherapy for lung sparing.
Due to the angulation of the breast board used for tangential breast irradiation, additional normal lung tissues are included in the supraclavicular field. This work investigates a method to reduce the lung volume and dose delivered during supraclavicular irradiation for breast cancer. Ten patients included for this retrospective study received chest wall and supraclavicular irradiation following radical surgery or breast-conserving surgery. Three-dimensional conformal radiation therapy plans were generated using the CMS XiO treatment planning system. The clinical target volume (CTV) of the supraclavicular irradiation is defined as the subcutaneous tissues from 0.5 cm under the anterior skin surface to a 3 cm depth. Only the ipsilateral lung is defined as the organ at risk. In the new method, the couch is rotated 90° and the supraclavicular field is tilted to maintain a normal incident angle to the breast board rather than the couch surface to spare more normal lung tissues. The absolute volume of the ipsilateral lung irradiated, and the volumes of lung tissues receiving 5 Gy and 20 Gy (V5 and V20) are analyzed. The new method can reduce the lung volume irradiated by the supraclavicular field significantly. For the ten patients investigated, only 5.3% of the ipsilateral lung is irradiated with the new method, while 14.9% of the ipsilateral lung is irradiated using the conventional method. Compared with the conventional method, the new method reduces V5 by 53.6% and V20 by 59.0%. Our new method does not alter the patient positioning for breast treatment but rotates the couch to deliver a tilted supraclavicular field to maintain adequate CTV coverage and spare more normal lung tissues. The results of this study demonstrated that our new method is effective, and that the reduction of normal lung tissue volume in the field is significant.


Eriocaulon buergerianum extract protects PC12 cells and neurons in zebrafish against 6-hydroxydopamine-induced damage.
Ericaulon buergerianum (Gujingcao) is an ophthalmic, anti-inflammatory and antimicrobial Chinese medicinal herb. This study aims to investigate the neuroprotective effects of Ericaulon buergerianum ethanol extract (EBE) and to elucidate its underlying action mechanism. The viability of dopaminergic (DA) neuron in zebrafish was examined by anti-tyrosine hydroxylase (TH) immunostaining. The locomotor activity of zebrafish was assessed with a digital video tracking system. The viability and cellular damage of the PC12 cells were determined by MTT and LDH assays respectively. The nuclear morphological changes in apoptotic cells were evaluated with DNA staining by Hoechst 33342 dye. Intracellular nitric oxide (NO) was quantified by DAF-FM diacetate staining. The expression of inducible nitric oxide synthase (iNOS) was determined by Western blot. EBE inhibited the 6-OHDA-induced decrease in total distance of movement in zebrafish. Pretreatments of EBE (25, 50, 100 and 200 μg/ml) increased the viability of 6-OHDA-damaged PC12 cells in a dose dependent manner. Protection against 6-OHDA-induced nuclear fragmentation and accumulation of apoptotic bodies was also observed in EBE pretreated cells. Anti-oxidative (inhibition of NO production and iNOS expression in PC12 cells in vitro) activities of EBE are related to its neuroprotective effects in 6-OHDA-induced DA neuron damage. EBE exhibited significant neuroprotective activities in zebrafish, including recovery of dopaminergic neuron loss caused by 6-OHDA in a dose-dependent manner in vivo, inhibition of 6-OHDA-induced decrease of total distance in movement in zebrafish. The iNOS-NO pathway may be involved.