Arms races with mitochondrial genome soft sweeps in a gynodioecious plant, Plantago lanceolata.
Biological situations involving conflict can create arms race situations with repeated fixations of different functional variants, producing selective sweeps and lowering neutral diversity in genome regions linked to the functional locus. However, they can sometimes lead to balancing selection, potentially creating long coalescent times for sites with functionally different variants, and, if recombination occurs rarely, for extended haplotypes carrying such variants. We tested between these possibilities in a gynodioecious plant, Plantago lanceolata, in which cytoplasmic male-sterility factors conflict with nuclear restorers of male fertility. We find low mitochondrial diversity, which does not support very long-term coexistence of highly diverged mitochondrial haplotypes. Interestingly, however, we found a derived haplotype that is associated with male fertility in a restricted geographic region, and that has fixed differences from the ancestral sequence in several genes, suggesting that it did not arise very recently. Taken together, the results suggest arms race events that involved "soft" selective sweeps involving a moderately old-established haplotype, consistent with the frequency fluctuations predicted by theoretical models of gynodioecy.


Low temperatures can promote cyanobacterial bloom formation by providing refuge from microbial antagonists.
Freshwater cyanobacteria are prone to a wide range of highly potent microbial antagonists. Most of these exploit their prey in a frequency-dependent manner and are therefore particularly well suited to prevent any accumulation of cyanobacteria. Mass developments of cyanobacteria, the so-called blooms, should therefore be rare events, which is in striking contrast to what we actually see in nature. Laboratory experiments of the present study showed that the temperature range 5.8-10 °C forms a thermal refuge, inside which the cyanobacterium Planktothrix can grow without being exploited by two otherwise highly potent microbial antagonists. In nature, access of Planktothrix to this refuge was associated with positive net growth and a high probability of bloom formation, confirming that refuge temperatures indeed allow Planktothrix to grow with a minimum of biomass loss caused by microbial antagonists. Contact to higher temperatures, in contrast, was associated with decreases in net growth rate and in probability of bloom formation, with population collapses and with the occurrence of parasite infection. This is in agreement with the finding of laboratory experiments that above 10 °C exploitation of Planktothrix by multiple microbes increases in a temperature-dependent manner. Taken together, above findings suggest that temperature modulates the microbial control of natural Planktothrix populations. Low temperatures form a thermal refuge that may promote Planktothrix bloom formation by shielding the cyanobacterium from otherwise highly potent microbial antagonists.


Polysaccharide Krestin enhances manganese superoxide dismutase activity and mRNA expression in mouse peritoneal macrophages.
Manganese superoxide dismutase (MnSOD), an inductive antioxidant enzyme, can protect cells from oxidative injury to the mitochondria. The elevation of MnSOD activity in cells can effectively prevent many diseases associated with oxidative stress. Polysaccharide Krestin (PSK), a kind of protein-bound polysaccharide extracted from Coriolus versicolor, is used as an immune response modifier in anti-tumor therapy. We have previously found that PSK could alleviate the oxidative injury that oxidized low density lipoprotein (Ox-LDL) brought to monocytes/macrophages, and therefore had some preventive or therapeutic effect on atherosclerosis. In order to find out if the effects of PSK were associated with the alteration ofantioxidant enzymes, we investigated its effect on MnSOD activity and gene expression in mouse peritoneal macrophages. The results showed that PSK could enhance SOD activity and increase the contents ofMnSOD mRNA in mouse peritoneal macrophages. Furthermore, the induction of MnSOD by PSK could be blocked by cycloheximide and actinomycin D.


The Chief Primary Care Medical Officer: Restoring Continuity.
The year 2016 marked the 20th anniversary of the hospitalist profession, with more than 50,000 physicians identifying as hospitalists. The Achilles heel of hospitalist medicine, however, is discontinuity. Despite many current payment and delivery systems rewarding this discontinuity and severing long-term relationships between patient and primary care teams at the hospital door, primary care does not stop being important when a person is admitted to the hospital. The notion of a broken primary care continuum is not an academic construct, it causes real harm to patients. As a step toward fixing the discontinuity in our health care systems, we propose that every hospital needs a Chief Primary Care Medical Officer (CPCMO), an expert in practice across the spectrum of care. The CPCMO can lead hospital efforts to create systems that ensure primary care's continuum is complete, while strengthening physician collaboration across specialties, and moving toward achieving the Quadruple Aim of enhancing patient experience, improving population health, reducing costs, and improving the work life of health care providers. For hospitals operating on value-based payment structures, anticipated improvement in measurable outcomes such as decreased length of stay, decreased readmission rates, improved transitions of care, improved patient satisfaction, improved access to primary care, and improved patient health, will enhance the rate of return on the hospital's investment. The speciality of family medicine should reevaluate our purpose, and reembrace our mission as personal physicians by championing the creation of Chief Primary Care Medical Officers.


Copeptin adds prognostic information after ischemic stroke: results from the CoRisk study.
To evaluate and validate the incremental value of copeptin in the prediction of outcome and complications as compared with established clinical variables. In this prospective, multicenter, cohort study, we measured copeptin in the emergency room within 24 hours from symptom onset in 783 patients with acute ischemic stroke. The 2 primary end points were unfavorable functional outcome (modified Rankin Scale score 3-6) and mortality within 90 days. Secondary end points were any of 5 prespecified complications during hospitalization. In multivariate analysis, higher copeptin independently predicted unfavorable outcome (adjusted odds ratio 2.17 for any 10-fold copeptin increase [95% confidence interval {CI}, 1.46-3.22], p < 0.001), mortality (adjusted hazard ratio 2.40 for any 10-fold copeptin increase [95% CI, 1.60-3.60], p < 0.001), and complications (adjusted odds ratio 1.93 for any 10-fold copeptin increase [95% CI, 1.33-2.80], p = 0.001). The discriminatory accuracy, calculated with the area under the receiver operating characteristic curve, improved significantly for all end points when adding copeptin to the NIH Stroke Scale score and the multivariate models. Moreover, the combination of copeptin with a validated score encompassing both the NIH Stroke Scale and age led to a net reclassification improvement of 11.8% for functional outcome and of 37.2% for mortality. In patients with ischemic stroke, copeptin is a validated blood marker that adds predictive information for functional outcome and mortality at 3 months beyond stroke severity and age. Copeptin seems to be a promising new blood marker for prediction of in-hospital complications.


Outcomes of Ethics Consultations in Adult ICUs: A Systematic Review and Meta-Analysis.
Clinical ethics consultation as a mechanism for supporting patients, family, and staff during ethically challenging situations has become standard of care. Despite this, there is a lack of consensus about the effectiveness of clinical ethics consultation consultation in the ICU. We performed a systematic review of outcomes associated with clinical ethics consultation within adult ICUs. We searched MEDLINE, PubMed, Cochrane CENTRAL, Embase, PsycINFO, and Cumulative Index to Nursing and Allied Health Literature from 1984 to May 2017. Two reviewers independently screened articles, assessed eligibility, extracted data, and assessed risk of bias using the Cochrane Collaboration Risk of Bias tool and the Newcastle-Ottawa Scale. Pooled estimates of effect were calculated where possible. We screened 3,970 abstracts and reviewed 325 full-text articles of which 16 met all eligibility criteria. We examined changes in processes and outcomes as a result of clinical ethics consultation in the ICU. Categories of outcomes included user perception, clinical decision, or conflict resolution and resource utilization. The use of clinical ethics consultation in the ICU was associated with positive user experience (383/435 found clinical ethics consultation helpful), although stress and disagreement with clinical ethics consultation recommendations was greater in a subset (113/431 surrogates and providers). Consensus for a clinical decision was more frequently achieved with clinical ethics consultation (odds ratio, 4.09; 95% CI, 1.01-16.55; p = 0.05). Clinical ethics consultation was associated with lower resource utilization including significantly decreased ICU length of stay (mean difference, -4.65 d; 95% CI, -8.86 to -0.44; p = 0.03). Our review identified outcome-based assessment as the predominant measure used to report effectiveness of clinical ethics consultation consultations. In particular, clinical ethics consultation decreased ICU length of stay and increased family and healthcare provider satisfaction. However, using outcome measures as the primary endpoint may not reflect the original intent of the clinical ethics consultation service. Based on our review, we propose a list of process measures that may better capture the key domains of a quality clinical ethics consultation.


General Concepts in Adult Congenital Heart Disease.
Congenital heart disease in adults (adult congenital heart disease) is a growing burden for healthcare systems. While infant mortality due to congenital heart disease in the last four decades decreased by almost 3-fold, adult congenital heart disease prevalence increased by more than 2-fold in United States. Adult congenital heart disease prevalence is expected to increase steadily until 2050 in projections. Adult congenital heart disease is a multifaceted problem with many dimensions. This manuscript aims to provide an overview of the common adult congenital heart diseases and summarize important points in management of these diseases with possible problems and complications that the patients and the physicians face.


Optimization of ozonation and peroxone process for simultaneous control of micropollutants and bromate in wastewater.
The study aims to simultaneously control micropollutants and bromate formations by using ozonation and peroxone process. The batch experiments were run with variations in specific ozone dose (SOD) and hydrogen peroxide-to-ozone (H2O2/O3) ratio. Based on the removal by ozonation and peroxone, micropollutants were categorized into three groups: non-reactive compounds (i.e. amidotrizoate), moderately reactive compounds (i.e. metoprolol, acesulfame potassium, bezafibrate, and benzotriazole), and highly reactive compounds (i.e. carbamazepine and diclofenac). For ozonation and peroxone process, the removals for highly reactive compounds and moderately reactive compounds were 82-99% and 29-99%, respectively. The removal of amidotrizoate was not observed in this study. The effect of ozonation on micropollutant removals was similar to the peroxone process. However, differences in bromate formation were observed. Bromate formation depended on the SOD, while addition of hydrogen peroxide suppressed the bromate formation. The peroxone process at the H2O2/O3 ratio of 0.3 was recommended to bromide-containing water below 100 µg·L-1 for simultaneous control of micropollutants and bromate. Enhancement in micropollutant removals, except for the non-reactive groups, was achieved with either higher SOD or the addition of hydrogen peroxide to ozonation. The micropollutant removal predicted from the second-order kinetic reaction with ozone and •OH exposures was higher than the observed data.


Digestive physiology of captive giant anteaters (Myrmecophaga tridactyla): determinants of faecal dry matter content.
Giant anteaters (Myrmecophaga tridactyla) are specialized insectivores and consume mainly ants and termites in the wild. In captivity, giant anteaters are either fed a complete diet, or a combination of a domestic carnivore diet with leaf eater pellets, or a traditional gruel-type diet. Soft faeces are a frequently encountered problem with this type of feeding. In the present study, we analysed diet and faeces composition, calculated digestibility and measured mean retention time on various diets in eight giant anteaters (total of n = 64 experiments). The results suggest that the digestive physiology of giant anteaters is similar to that of domestic dogs and cats in terms of nutrient digestibility and digesta retention. When testing correlations between faecal dry matter content and other variables, no relationship with dietary crude fibre content or mean digesta retention time could be detected. However, acid insoluble ash intake was significantly and positively correlated with faecal dry matter content. The amount of acid insoluble ash excreted with the faeces was higher than that ingested with the diet offered, indicating that the giant anteaters ingested soil from their enclosure of up to 93 g per day. This finding is consistent with observation of faeces of wild giant anteaters that contain soil or sand most likely due to indiscriminate feeding. It also corresponds to reports that indigestible materials such as peat, soil, chitin or cellulose contribute to a firmer faecal consistency in various carnivore species. Therefore, offering giant anteaters the opportunity to voluntarily ingest soil from their enclosure might be beneficial.


Long Receptor Residence Time of C26 Contributes to Super Agonist Activity at the Human β2 Adrenoceptor.
Super agonists produce greater functional responses than endogenous agonists in the same assay, and their unique pharmacology is the subject of increasing interest and debate. We propose that receptor residence time and the duration of receptor signaling contribute to the pharmacology of super agonism. We have further characterized the novel β2 adrenoceptor agonist C26 (7-[(R)-2-((1R,2R)-2-benzyloxycyclopentylamino)-1-hydroxyethyl]-4-hydroxybenzothiazolone), which displays higher intrinsic activity than the endogenous ligand adrenaline in cAMP accumulation, β-arrestin-2 recruitment, and receptor internalization assays. C26 recruited β-arrestin-2, and internalized the Green Fluorescent Protein (GFP)-taggedβ2 adrenoceptor at a slow rate, with half-life (t1/2) values of 0.78 ± 0.1 and 0.78 ± 0.04 hours, respectively. This was compared with 0.31 ± 0.04 and 0.34 ± 0.01 hours for adrenaline-mediated β-arrestin-2 recruitment and GFP-β2 internalization, respectively. The slower rate for C26 resulted in levels of β-arrestin-2 recruitment increasing up to 4-hour agonist incubation, at which point the intrinsic activity was determined to be 124.3 ± 0.77% of the adrenaline response. In addition to slow functional kinetics, C26 displayed high affinity with extremely slow receptor dissociation kinetics, giving a receptor residence half-life of 32.7 minutes at 37°C, which represents the slowest dissociation rate we have observed for any β2 adrenoceptor agonist tested to date. In conclusion, we propose that the gradual accumulation of long-lived active receptor complexes contributes to the increased intrinsic activity of C26 over time. This highlights the need to consider the temporal aspects of agonist binding and signaling when characterizing ligands as super agonists.


In situ structure and dynamics of DNA origami determined through molecular dynamics simulations.
The DNA origami method permits folding of long single-stranded DNA into complex 3D structures with subnanometer precision. Transmission electron microscopy, atomic force microscopy, and recently cryo-EM tomography have been used to characterize the properties of such DNA origami objects, however their microscopic structures and dynamics have remained unknown. Here, we report the results of all-atom molecular dynamics simulations that characterized the structural and mechanical properties of DNA origami objects in unprecedented microscopic detail. When simulated in an aqueous environment, the structures of DNA origami objects depart from their idealized targets as a result of steric, electrostatic, and solvent-mediated forces. Whereas the global structural features of such relaxed conformations conform to the target designs, local deformations are abundant and vary in magnitude along the structures. In contrast to their free-solution conformation, the Holliday junctions in the DNA origami structures adopt a left-handed antiparallel conformation. We find the DNA origami structures undergo considerable temporal fluctuations on both local and global scales. Analysis of such structural fluctuations reveals the local mechanical properties of the DNA origami objects. The lattice type of the structures considerably affects global mechanical properties such as bending rigidity. Our study demonstrates the potential of all-atom molecular dynamics simulations to play a considerable role in future development of the DNA origami field by providing accurate, quantitative assessment of local and global structural and mechanical properties of DNA origami objects.


Phenotype harmonization and cross-study collaboration in GWAS consortia: the GENEVA experience.
Genome-wide association study (GWAS) consortia and collaborations formed to detect genetic loci for common phenotypes or investigate gene-environment (G*E) interactions are increasingly common. While these consortia effectively increase sample size, phenotype heterogeneity across studies represents a major obstacle that limits successful identification of these associations. Investigators are faced with the challenge of how to harmonize previously collected phenotype data obtained using different data collection instruments which cover topics in varying degrees of detail and over diverse time frames. This process has not been described in detail. We describe here some of the strategies and pitfalls associated with combining phenotype data from varying studies. Using the Gene Environment Association Studies (GENEVA) multi-site GWAS consortium as an example, this paper provides an illustration to guide GWAS consortia through the process of phenotype harmonization and describes key issues that arise when sharing data across disparate studies. GENEVA is unusual in the diversity of disease endpoints and so the issues it faces as its participating studies share data will be informative for many collaborations. Phenotype harmonization requires identifying common phenotypes, determining the feasibility of cross-study analysis for each, preparing common definitions, and applying appropriate algorithms. Other issues to be considered include genotyping timeframes, coordination of parallel efforts by other collaborative groups, analytic approaches, and imputation of genotype data. GENEVA's harmonization efforts and policy of promoting data sharing and collaboration, not only within GENEVA but also with outside collaborations, can provide important guidance to ongoing and new consortia.


Silicon-based optoelectronic integrated circuit for label-free bio/chemical sensor.
We demonstrate a silicon-based optoelectronic integrated circuit (OEIC) for label-free bio/chemical sensing application. Such on-chip OEIC sensor system consists of optical grating couplers for vertical light coupling into silicon waveguides, a thermal-tunable microring as a tunable filter, an exposed microring as an optical label-free sensor, and a Ge photodetector for a direct electrical readout. Different from the conventional wavelength-scanning method, we adopt low-cost broadband ASE light source, together with the on-chip tunable filter to generate sliced light source. The effective refractive index change of the sensing microring induced by the sensing target is traced by scanning the supplied electrical power applied onto the tracing microring, and the detected electrical signal is read out by the Ge photodetector. For bulk refractive index sensing, we demonstrate using such OEIC sensing system with a sensitivity of ~15 mW/RIU and a detection limit of 3.9 μ-RIU, while for surface sensing of biotin-streptavidin, we obtain a surface mass sensitivity of S(m) = ~192 µW/ng·mm(-2) and a surface detection limit of 0.3 pg/mm(2). The presented OEIC sensing system is suitable for point-of-care applications.


Vascular gene expression: a hypothesis.
The phloem is the conduit through which photoassimilates are distributed from autotrophic to heterotrophic tissues and is involved in the distribution of signaling molecules that coordinate plant growth and responses to the environment. Phloem function depends on the coordinate expression of a large array of genes. We have previously identified conserved motifs in upstream regions of the Arabidopsis genes, encoding the homologs of pumpkin phloem sap mRNAs, displaying expression in vascular tissues. This tissue-specific expression in Arabidopsis is predicted by the overrepresentation of GA/CT-rich motifs in gene promoters. In this work we have searched for common motifs in upstream regions of the homologous genes from plants considered to possess a "primitive" vascular tissue (a lycophyte), as well as from others that lack a true vascular tissue (a bryophyte), and finally from chlorophytes. Both lycophyte and bryophyte display motifs similar to those found in Arabidopsis with a significantly low E-value, while the chlorophytes showed either a different conserved motif or no conserved motif at all. These results suggest that these same genes are expressed coordinately in non-vascular plants; this coordinate expression may have been one of the prerequisites for the development of conducting tissues in plants. We have also analyzed the phylogeny of conserved proteins that may be involved in phloem function and development. The presence of CmPP16, APL, FT, and YDA in chlorophytes suggests the recruitment of ancient regulatory networks for the development of the vascular tissue during evolution while OPS is a novel protein specific to vascular plants.


[Knowledge about radon and its associated risk perception in France].
Radon exposure is a major environmental risk in health. It remains badly known by the general population. It is the second cause of lung cancer, after tobacco smoking. The aim of this cross-sectional general population survey was to describe radon exposure risk knowledge and the socioeconomic factors related to this knowledge. The Cancer Barometer survey 2010 questioned the French population about its knowledge of radon as such and as health risk factor. This survey was a two-stage random sampling with computer-assisted telephone interview that was performed from April 3, 2010 to August 7, 2010 on a sample of 3,359 people aged 15 to 75 years old. Among people aged 15 to 75 years old, only one in five knows that radon is a natural gas coming from the ground. This knowledge is more frequent among people living in an area that is directly concerned by radon, among men and increases with age, with the level of education and the level of income. Radon risk remains still widely underestimated by the general public, including in areas concerned by this risk. When people were confronted with radon exposure, few intended to remedy by improving their home. The success of prevention initiatives implies the support and the collaboration of various national and local actors. To improve their impact for the prevention of lung cancers, it could be more effective to couple these actions with prevention messages on tobacco.


Delay discounting, genetic sensitivity, and leukocyte telomere length.
In a graying world, there is an increasing interest in correlates of aging, especially those found in early life. Leukocyte telomere length (LTL) is an emerging marker of aging at the cellular level, but little is known regarding its link with poor decision making that often entails being overly impatient. Here we investigate the relationship between LTL and the degree of impatience, which is measured in the laboratory using an incentivized delay discounting task. In a sample of 1,158 Han Chinese undergraduates, we observe that steeper delay discounting, indexing higher degree of impatience, is negatively associated with LTL. The relationship is robust after controlling for health-related variables, as well as risk attitude-another important determinant of decision making. LTL in females is more sensitive to impatience than in males. We then asked if genes possibly modulate the effect of impatient behavior on LTL. The oxytocin receptor gene (OXTR) polymorphism rs53576, which has figured prominently in investigations of social cognition and psychological resources, and the estrogen receptor β gene (ESR2) polymorphism rs2978381, one of two gonadal sex hormone genes, significantly mitigate the negative effect of impatience on cellular aging in females. The current results contribute to understanding the relationship between preferences in decision making, particularly impatience, and cellular aging, for the first time to our knowledge. Notably, oxytocin and estrogen receptor polymorphisms temper accelerated cellular aging in young females who tend to make impatient choices.


Cholangiopancreatoscopy and endoscopic ultrasound for indeterminate pancreaticobiliary pathology.
Techniques to confirm suspected pancreaticobiliary (PB) malignancy when index sampling is non-diagnostic include cholangiopancreatoscopy (CP) and endoscopic ultrasound (EUS). However, comparative data are lacking. The purpose of this study was to compare the yield of EUS and CP for the diagnosis of PB pathology. Consecutive patients with indeterminate PB pathology who underwent both CP and EUS within 3 months of each other were retrospectively identified. For CP, tissue sampling included biopsy under direct inspection (cholangioscopy-directed biopsy), biopsy following CP with fluoroscopic guidance (cholangioscopy-assisted biopsy), or brush cytology. For EUS-FNA, lesions included ductal strictures or hypoechoic masses. A comparison of operating characteristics between CP and EUS utilizing tissue confirmation or 12-month clinical course consistent with either benign or malignant disease was performed. Between February 2000 and June 2007, 66 (33 males, 33 females, median age 64.5) patients with indeterminate PB pathology who had undergone both CP and EUS within 3 months of each other were included. Lesions amenable to sampling were noted in 59 CP and 50 EUS patients. On follow-up, 39 patients had neoplasia and 27 were benign. The sensitivity/specificity for the diagnosis of neoplasia for CP and EUS was 48.7/96.3 % and 33.3/96.3 %, respectively (comparison of sensitivities, P = 0.183). The combined (CP and EUS) sensitivity/specificity was 66.7/96.3 % (P = 0.0064 and P = 0.0001 comparing combined sensitivity vs. sensitivity of either CP alone or EUS alone, respectively). In patients who undergo both EUS and CP for indeterminate PB pathology, the combined yield of EUS and CP to detect neoplasia appears to be higher than either examination alone.


The role of TNF related apoptosis-inducing ligand in neurodegenerative diseases.
A hallmark of all forms of neurodegenerative diseases is impairment of neuronal functions, and in many cases neuronal cell death. Although the etiology of neurodegenerative diseases may be distinct, different diseases display a similar pathogenesis, for example abnormal immunity within the central nervous system (CNS), activation of macrophage/microglia and the involvement of proinflammatory cytokines. Recent studies show that neurons in a neurodegenerative state undergo a highly regulated programmed cell death, also called apoptosis. TNF-related apoptosis-inducing ligand (TRAIL), a member of the TNF family, has been shown to be involved in apoptosis during many diseases. As one member of a death ligand family, TRAIL was originally thought to target only tumor cells and was not present in CNS. However, recent data showed that TRAIL was unregulated in HIV-1-infected and immune-activated macrophages, a major disease inducing cell during HIV-1-associated dementia (HAD). TRAIL is also induced on neuron by beta-amyloid protein, an important pathogen for Alzheimer's disease. In this review, we summarize the possible common aspects that TRAIL involved those neurodegenerative diseases, TRAIL induced apoptosis signaling in the CNS cells, and specific role of TRAIL in individual diseases.


Activation of the peroxisome proliferator-activated receptor alpha protects against myocardial ischaemic injury and improves endothelial vasodilatation.
The peroxisome proliferator-activated receptor alpha (PPARalpha) plays an important role in the metabolism of lipoproteins and fatty acids, and seems to protect against the development of atherosclerosis. To evaluate the possible protective role of PPARalpha on cardiovascular function, the effect of the PPARalpha agonist, fenofibrate was assessed with respect to ischaemia/reperfusion injury and endothelial function in mice. Fenofibrate treatment reduces myocardial infarction size and improves post-ischaemic contractile dysfunction. Hearts from PPARalpha null mice exhibit increased susceptibility to ischaemic damages and were refractory to protection by fenofibrate treatment suggesting that the beneficial effects of fenofibrate were mediated via PPARalpha. Furthermore, fenofibrate improves endothelium- and nitric oxide-mediated vasodilatation in aorta and mesenteric vascular bed. A decreased inhibitory effect of reactive oxygen species in the vessel wall accounts for enhanced endothelial vasodilatation. However, the latter cannot be explained by an increase in nitric oxide synthase expression nor by an increase sensitivity of the arteries to nitric oxide. Altogether the present data suggest that fenofibrate exerts cardioprotective effect against ischaemia and improves nitric oxide-mediated response probably by enhancing antioxidant capacity of the vessel wall. These data underscore new therapeutic perspectives for PPARalpha agonists in ischaemic myocardial injury and in cardiovascular diseases associated with endothelial dysfunction.


Thyrotrophin-releasing hormone (TRH) stimulation test in manic-depressive illness.
The thyrotrophin (TSH) response to thyrotrophin-releasing hormone (TRH 200 microgram intravenously was studied in 19 patients with unipolar depression, 12 with bipolar depression, 14 with mania, and 5 with mixed manic-depressive illness. The TSH responses were decreased in all of these affective disorders compared to those found in 10 patients with reactive depression, 5 with reactive paranoid psychosis, 14 with neurotic depression, and 60 controls. The decrease of the TSH response in manics could not be accounted for by the effects of neuroleptic drugs. The TSH response in the groups with reactive depression, reactive paranoid psychosis, and neurotic depression, respectively, did not differ significantly from that found in controls. With the exception of a decrease in serum T3 level and free T3 index in the manics, no significant differences in serum T3 level or in free T3 and T4 indexes were found between the groups. Changes found in serum T4 level were due to changes in the thyroxine-binding proteins.


A systematic review of vertigo in primary care.
The symptom of vertigo is usually managed in primary care without further referral. This review examines the evidence on which general practitioners can base clinical diagnosis and management of this relatively common complaint. Research in this area has in the main been from secondary and tertiary centres and has been of variable quality. Indications are that the conditions that present in general practice are most likely to be benign positional vertigo, acute vestibular neuronitis, and Ménière's disease; however, vascular incidents and neurological causes, such as multiple sclerosis, must be kept in mind. An important practice point is that vestibular sedatives are not recommended on a prolonged basis for any type of vertigo. There is a need for basic epidemiological and clinical management research of vertigo in general practice.


Platelet-free calcium and vascular calcium uptake in ethanol-induced hypertensive rats.
This study examined the effect of moderate ethanol intake on systolic blood pressure, platelet cytosolic free calcium, aortic calcium, and rubidium-86 uptake in Wistar-Kyoto rats. Twelve Wistar-Kyoto rats, aged 6 weeks, were given 5% ethanol in drinking water the first week followed by 10% ethanol in drinking water for the next 6 weeks. Twelve control animals were given regular tap water. Systolic blood pressure in the ethanol-treated rats was significantly higher (p less than 0.05) than that in controls after 1 week and remained higher throughout the study. At 13 weeks of age, platelet cytosolic free calcium and calcium uptake by aortas were significantly higher (p less than 0.001) in ethanol-treated animals as compared with those in controls. Ethanol intake did not affect aortic ouabain-sensitive 86Rb uptake. The in vitro effect of ethanol on calcium-45 and 86Rb uptake was also investigated in aortas of untreated Wistar-Kyoto rats at 13 weeks of age. In vitro ethanol (2.5-20 mmols/l) did not significantly affect 45Ca and 86Rb uptake in rat aortas. The increases in systolic blood pressure, platelet cytosolic free calcium, and vascular calcium uptake suggest that increases in cytosolic free calcium and calcium uptake mechanisms are associated with ethanol-induced hypertension.


Rat models of acute inflammation: a randomized controlled study on the effects of homeopathic remedies.
One of the cardinal principles of homeopathy is the "law of similarities", according to which patients can be treated by administering substances which, when tested in healthy subjects, cause symptoms that are similar to those presented by the patients themselves. Over the last few years, there has been an increase in the number of pre-clinical (in vitro and animal) studies aimed at evaluating the pharmacological activity or efficacy of some homeopathic remedies under potentially reproducible conditions. However, in addition to some contradictory results, these studies have also highlighted a series of methodological difficulties.The present study was designed to explore the possibility to test in a controlled way the effects of homeopathic remedies on two known experimental models of acute inflammation in the rat. To this aim, the study considered six different remedies indicated by homeopathic practice for this type of symptom in two experimental edema models (carrageenan- and autologous blood-induced edema), using two treatment administration routes (sub-plantar injection and oral administration). In a first phase, the different remedies were tested in the four experimental conditions, following a single-blind (measurement) procedure. In a second phase, some of the remedies (in the same and in different dilutions) were tested by oral administration in the carrageenan-induced edema, under double-blind (treatment administration and measurement) and fully randomized conditions. Seven-hundred-twenty male Sprague Dawley rats weighing 170-180 g were used. Six homeopathic remedies (Arnica montana D4, Apis mellifica D4, D30, Atropa belladonna D4, Hamamelis virginiana D4, Lachesis D6, D30, Phosphorus D6, D30), saline and indomethacin were tested. Edema was measured using a water-based plethysmometer, before and at different times after edema induction. Data were analyzed by ANOVA and Student t test. In the first phase of experiments, some statistically significant effects of homeopathic remedies (Apis, Lachesis and Phosporus) were observed (the reduction in paw volume increase ranging from 10% to 28% at different times since edema induction). In the second phase of experiments, the effects of homeopathic remedies were not confirmed. On the contrary, the unblinded standard allopathic drug indomethacin exhibited its anti-inflammatory effect in both experimental phases (the reduction in paw volume increase ranging from 14% to 40% in the first phase, and from 18% to 38% in the second phase of experiments). The discrepancies between single-blind and double-blind methods in animal pharmacological research are noteworthy and should be better investigated, also in non-homeopathic research.


Towards a more desirable dry powder inhaler formulation: large spray-dried mannitol microspheres outperform small microspheres.
To investigate, for the first time, the performance of a dry powder inhaler (DPI, Aerolizer(®)) in the case of a model drug (i.e. albuterol sulphate) formulated with spray dried mannitol carrier particles with homogeneous shape and solid-state form but different sizes. Spray dried mannitol (SDM) particles were characterized in terms of size, surface area, morphology, water content, solid-state, density and electrostatic charge by a novel approach. DPI formulations composed of SDM and albuterol sulphate (AS) were prepared and evaluated in terms of drug content homogeneity and in vitro aerosolization performance. All SDM particles generated similar fine particle fractions of AS. Formulations consisting of larger SDM particles demonstrated better drug content homogeneity, reduced amounts of drug loss and reduced oropharyngeal deposition. Comparing different SDM products demonstrated that SDM powders with relatively poorer flowability, wider size distributions and higher charge density generated DPI formulations with poorer drug content homogeneity and deposited higher amount of drug on the inhaler, mouthpiece adaptor and throat. DPI formulation total desirability increased linearly with the mean diameter of SDM. Particle shape and solid-state form of mannitol could dominate over carrier size, bulk density, flowability and charge in terms of determining the aerosolization behaviour of AS formulated with mannitol carrier, at least within the experimental protocols applied in the present study.


Electron Doping of the Parent Cuprate La_{2}CuO_{4} without Cation Substitution.
In the cuprates, carrier doping of the Mott insulating parent state is necessary to realize superconductivity as well as a number of other exotic states involving charge or spin density waves. Cation substitution is the primary method for doping carriers into these compounds, and is the only known method for electron doping in these materials. Here, we report electron doping without cation substitution in epitaxially stabilized thin films of La_{2}CuO_{4} grown via molecular-beam epitaxy. We use angle-resolved photoemission spectroscopy to directly measure their electronic structure and conclusively determine that these compounds are electron doped with a carrier concentration of 0.09±0.02 e^{-}/Cu. We propose that intrinsic defects, most likely oxygen vacancies, are the sources of doped electrons in these materials. Our results suggest a new approach to electron doping in the cuprates, one which could lead to a more detailed experimental understanding of their properties.


Precise colocalization of interacting structural and pigmentary elements generates extensive color pattern variation in Phelsuma lizards.
Color traits in animals play crucial roles in thermoregulation, photoprotection, camouflage, and visual communication, and are amenable to objective quantification and modeling. However, the extensive variation in non-melanic pigments and structural colors in squamate reptiles has been largely disregarded. Here, we used an integrated approach to investigate the morphological basis and physical mechanisms generating variation in color traits in tropical day geckos of the genus Phelsuma. Combining histology, optics, mass spectrometry, and UV and Raman spectroscopy, we found that the extensive variation in color patterns within and among Phelsuma species is generated by complex interactions between, on the one hand, chromatophores containing yellow/red pteridine pigments and, on the other hand, iridophores producing structural color by constructive interference of light with guanine nanocrystals. More specifically, we show that 1) the hue of the vivid dorsolateral skin is modulated both by variation in geometry of structural, highly ordered narrowband reflectors, and by the presence of yellow pigments, and 2) that the reflectivity of the white belly and of dorsolateral pigmentary red marks, is increased by underlying structural disorganized broadband reflectors. Most importantly, these interactions require precise colocalization of yellow and red chromatophores with different types of iridophores, characterized by ordered and disordered nanocrystals, respectively. We validated these results through numerical simulations combining pigmentary components with a multilayer interferential optical model. Finally, we show that melanophores form dark lateral patterns but do not significantly contribute to variation in blue/green or red coloration, and that changes in the pH or redox state of pigments provide yet another source of color variation in squamates. Precisely colocalized interacting pigmentary and structural elements generate extensive variation in lizard color patterns. Our results indicate the need to identify the developmental mechanisms responsible for the control of the size, shape, and orientation of nanocrystals, and the superposition of specific chromatophore types. This study opens up new perspectives on Phelsuma lizards as models in evolutionary developmental biology.


Optically active quantum dots in monolayer WSe2.
Semiconductor quantum dots have emerged as promising candidates for the implementation of quantum information processing, because they allow for a quantum interface between stationary spin qubits and propagating single photons. In the meantime, transition-metal dichalcogenide monolayers have moved to the forefront of solid-state research due to their unique band structure featuring a large bandgap with degenerate valleys and non-zero Berry curvature. Here, we report the observation of zero-dimensional anharmonic quantum emitters, which we refer to as quantum dots, in monolayer tungsten diselenide, with an energy that is 20-100 meV lower than that of two-dimensional excitons. Photon antibunching in second-order photon correlations unequivocally demonstrates the zero-dimensional anharmonic nature of these quantum emitters. The strong anisotropic magnetic response of the spatially localized emission peaks strongly indicates that radiative recombination stems from localized excitons that inherit their electronic properties from the host transition-metal dichalcogenide. The large ∼1 meV zero-field splitting shows that the quantum dots have singlet ground states and an anisotropic confinement that is most probably induced by impurities or defects. The possibility of achieving electrical control in van der Waals heterostructures and to exploit the spin-valley degree of freedom renders transition-metal-dichalcogenide quantum dots interesting for quantum information processing.


Low-Temperature Solution-Processed Kesterite Solar Cell Based on in Situ Deposition of Ultrathin Absorber Layer.
The production of high-performance, solution-processed kesterite Cu2ZnSn(Sx,Se1-x)4 (CZTSSe) solar cells typically relies on high-temperature crystallization processes in chalcogen-containing atmosphere and often on the use of environmentally harmful solvents, which could hinder the widespread adoption of this technology. We report a method for processing selenium free Cu2ZnSnS4 (CZTS) solar cells based on a short annealing step at temperatures as low as 350 °C using a molecular based precursor, fully avoiding highly toxic solvents and high-temperature sulfurization. We show that a simple device structure consisting of ITO/CZTS/CdS/Al and comprising an extremely thin absorber layer (∼110 nm) achieves a current density of 8.6 mA/cm(2). Over the course of 400 days under ambient conditions encapsulated devices retain close to 100% of their original efficiency. Using impedance spectroscopy and photoinduced charge carrier extraction by linearly increasing voltage (photo-CELIV), we demonstrate that reduced charge carrier mobility is one limiting parameter of low-temperature CZTS photovoltaics. These results may inform less energy demanding strategies for the production of CZTS optoelectronic layers compatible with large-scale processing techniques.


Sand Flies and Their Control Methods.
The main aim of managing arthropod vectors that carry the disease agents is interrupting the infection cycle. Therefore, the management of the disease implies that all precautions related to all elements (i.e., human, arthropod vector, and reservoir) in the infection cycle need to be taken. There are important points that need to be considered while dealing with sand flies (Diptera: Psychodidae: Phlebotominae), which in many regions worldwide, particularly in tropical and subtropical areas, are vectors of diseases such as leishmaniasis and sand fly fever and are the arthropods of the infection cycle. Because the larval control of the sand flies is very difficult and almost impossible, the management is mainly conducted for the adults. The most effective strategy for reducing both sand fly fever and leishmaniasis is managing sand flies, particularly in areas where humans are located. In this review, the morphology, biology, and taxonomy of sand flies; the integrated fighting and management methods such as insecticide-impregnated bed nets and use of curtains, zooprophylaxis, indoor and outdoor residual applications, larvicides, repellents, and insecticide-impregnated dog collars; and data regarding many issues such as insecticide resistance in sand flies have been emphasized on in the review.


Kinetics of Local and Systemic Leucocyte and Cytokine Reaction of Calves to Intrabronchial Infection with Chlamydia psittaci.
Infection of cattle with chlamydiae is ubiquitous and, even in the absence of clinical sequeleae, has a quantifiable negative impact on livestock productivity. Despite recent progress, our knowledge about immune response mechanisms capable of counteracting the infection and preventing its detrimental effects is still limited. A well-established model of bovine acute respiratory Chlamydia (C.) psittaci infection was used here to characterize the kinetics of the local and systemic immune reactions in calves. In the course of two weeks following inoculation, leukocyte surface marker expression was monitored by flow cytometry in blood and bronchoalveolar lavage fluid (BALF). Immune-related protein and receptor transcription were determined by quantitative real-time reverse transcription PCR in blood, BALF and lung tissue. An early increase of IL2RA, IL10 and HSPA1A mRNA expressions was followed by a rise of lymphocytes, monocytes, and granulocytes exhibiting activated phenotypes in blood. Monocytes showed elevated expression rates of CD11b, CD14 and MHC class II. The rates of CD62L expression on CD8hi T cells in blood and on CD4+ T cells in BALF were also augmented and peaked between 2 and 4 dpi. Notably, CD25 antigen expression was significantly elevated, not only on CD8dim/CD62L+ and CD8-/CD62L+ cells in blood, but also on granulocytes in blood and BALF between 2-3 dpi. From 4 dpi onwards, changes declined and the calves recovered from the infection until 10 dpi. The findings highlight the effectiveness of rapid local and systemic immune reaction and indicate activated T cells, monocytes and granulocytes being essential for rapid eradication of the C. psittaci infection.


Fear of Dementia and the Obligation to Provide Aggregate Research Results to Study Participants.
A general obligation to make aggregate research results available to participants has been widely supported in the bioethics literature. However, dementia research presents several challenges to this perspective, particularly because of the fear associated with developing dementia. The authors argue that considerations of respect for persons, beneficence, and justice fail to justify an obligation to make aggregate research results available to participants in dementia research. Nevertheless, there are positive reasons in favor of making aggregate research results available; when the decision is made to do so, it is critical that a clear strategy for communicating results is developed, including what support will be provided to participants receiving aggregate research results.


Health system and community level interventions for improving antenatal care coverage and health outcomes.
The World Health Organization (WHO) recommends at least four antenatal care (ANC) visits for all pregnant women. Almost half of pregnant women worldwide, and especially in developing countries do not receive this amount of care. Poor attendance of ANC is associated with delivery of low birthweight babies and more neonatal deaths. ANC may include education on nutrition, potential problems with pregnancy or childbirth, child care and prevention or detection of disease during pregnancy.This review focused on community-based interventions and health systems-related interventions. To assess the effects of health system and community interventions for improving coverage of antenatal care and other perinatal health outcomes. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (7 June 2015) and reference lists of retrieved studies. We included randomised controlled trials (RCTs), quasi-randomised trials and cluster-randomised trials. Trials of any interventions to improve ANC coverage were eligible for inclusion. Trials were also eligible if they targeted specific and related outcomes, such as maternal or perinatal death, but also reported ANC coverage. Two review authors independently assessed trials for inclusion and risk of bias, extracted data and checked them for accuracy. We included 34 trials involving approximately 400,000 women. Some trials tested community-based interventions to improve uptake of antenatal care (media campaigns, education or financial incentives for pregnant women), while other trials looked at health systems interventions (home visits for pregnant women or equipment for clinics). Most trials took place in low- and middle-income countries, and 29 of the 34 trials used a cluster-randomised design. We assessed 30 of the 34 trials as of low or unclear overall risk of bias. Comparison 1: One intervention versus no interventionWe found marginal improvements in ANC coverage of at least four visits (average odds ratio (OR) 1.11, 95% confidence interval (CI) 1.01 to 1.22; participants = 45,022; studies = 10; Heterogeneity: Tau² = 0.01; I² = 52%; high quality evidence). Sensitivity analysis with a more conservative intra-cluster correlation co-efficient (ICC) gave similar marginal results. Excluding one study at high risk of bias shifted the marginal pooled estimate towards no effect. There was no effect on pregnancy-related deaths (average OR 0.69, 95% CI 0.45 to 1.08; participants = 114,930; studies = 10; Heterogeneity: Tau² = 0.00; I² = 0%; low quality evidence), perinatal mortality (average OR 0.98, 95% CI 0.90 to 1.07; studies = 15; Heterogeneity: Tau² = 0.01; I² = 58%; moderate quality evidence) or low birthweight (average OR 0.94, 95% CI 0.82 to 1.06; studies = five; Heterogeneity: Tau² = 0.00; I² = 5%; high quality evidence). Single interventions led to marginal improvements in the number of women who delivered in health facilities (average OR 1.08, 95% CI 1.02 to 1.15; studies = 10; Heterogeneity: Tau² = 0.00; I² = 0%; high quality evidence), and in the proportion of women who had at least one ANC visit (average OR 1.68, 95% CI 1.02 to 2.79; studies = six; Heterogeneity: Tau² = 0.24; I² = 76%; moderate quality evidence). Results for ANC coverage (at least four and at least one visit) and for perinatal mortality had substantial statistical heterogeneity. Single interventions did not improve the proportion of women receiving tetanus protection (average OR 1.03, 95% CI 0.92 to 1.15; studies = 8; Heterogeneity: Tau² = 0.01; I² = 57%). No study reported onintermittent prophylactic treatment for malaria. Comparison 2: Two or more interventions versus no interventionWe found no improvements in ANC coverage of four or more visits (average OR 1.48, 95% CI 0.99 to 2.21; participants = 7840; studies = six; Heterogeneity: Tau² = 0.10; I² = 48%; low quality evidence) or pregnancy-related deaths (average OR 0.70, 95% CI 0.39 to 1.26; participants = 13,756; studies = three; Heterogeneity: Tau² = 0.00; I² = 0%; moderate quality evidence). However, combined interventions led to improvements in ANC coverage of at least one visit (average OR 1.79, 95% CI 1.47 to 2.17; studies = five; Heterogeneity: Tau² = 0.00; I² = 0%; moderate quality evidence), perinatal mortality (average OR 0.74, 95% CI 0.57 to 0.95; studies = five; Heterogeneity: Tau² = 0.06; I² = 83%; moderate quality evidence) and low birthweight (average OR 0.61, 95% CI 0.46 to 0.80; studies = two; Heterogeneity: Tau² = 0.00; I² = 0%; moderate quality evidence). Meta-analyses for both ANC coverage four or more visits and perinatal mortality had substantial statistical heterogeneity. Combined interventions improved the proportion of women who had tetanus protection (average OR 1.48, 95% CI 1.18 to 1.87; studies = 3; Heterogeneity: Tau² = 0.01; I² = 33%). No trial in this comparison reported on intermittent prophylactic treatment for malaria. Comparison 3: Two interventions compared head to head. No trials found. Comparison 4: One intervention versus a combination of interventionsThere was no difference in ANC coverage (four or more visits and at least one visit), pregnancy-related deaths, deliveries in a health facility or perinatal mortality. No trials in this comparison reported on low birthweight orintermittent prophylactic treatment of malaria. Implications for practice - Single interventions may improve ANC coverage (at least one visit and four or more visits) and deliveries in health facilities. Combined interventions may improve ANC coverage (at least one visit), reduce perinatal mortality and reduce the occurrence of low birthweight. The effects of the interventions are unrelated to whether they are community or health system interventions. Implications for research - More details should be provided in reporting numbers of events, group totals and the ICCs used to adjust for cluster effects. Outcomes should be reported uniformly so that they are comparable to commonly-used population indicators. We recommend further cluster-RCTs of pregnant women and women in their reproductive years, using combinations of interventions and looking at outcomes that are important to pregnant women, such as maternal and perinatal morbidity and mortality, alongside the explanatory outcomes along the pathway of care: ANC coverage, the services provided during ANC and deliveries in health facilities.


Two new species of dwarf rainbowfishes (Atheriniformes: Melanotaeniidae) from northern Australia and southern New Guinea.
The 'maccullochi species group' of rainbowfishes are small and distinctly patterned freshwater fishes of streams and swamps, comprising around eight species. The species from which the group bears its name, Melanotaenia maccullochi Ogilby, 1915, has been thought to comprise three forms occurring in distinct geographic areas, and recent mitochondrial genetic data provides matching patterns of likely inter-specific divergence. Here we undertake a detailed investigation of the taxonomic status of M. maccullochi using a combined lines of evidence approach incorporating multiple nuclear genetic markers (55 allozyme loci), mitochondrial DNA sequence data (1141 bp cytochrome b) and morphology (examination of a suite of 38 morphometric and meristic characters). As all three datasets provide support for a three-way split, we accordingly describe two new species and redescribe M. maccullochi sensu stricto. McCulloch's Rainbowfish M. maccullochi, a species with brown body stripes and red fins occurs in northeast Queensland and is redescribed based on 338 specimens, 13.1-53.0 mm SL. This species was one of the first rainbowfishes to become known in the aquarium hobby. A second form with darker stripes on a contrasting light white-grey body and with distinct sub-marginal black bands in the dorsal and anal fins, distributed across northern and eastern Cape York Peninsula, Torres Strait and southern central New Guinea, is described as Sahul Rainbowfish M. sahulensis sp. nov. based on 267 specimens, 13.4-48.4 mm SL. A diminutive and well geographically isolated form occurring below the escarpment of the Tabletop Range in Litchfield National Park, Northern Territory possessing a more prominent and purplish mid-lateral stripe, is described as the Little Rainbowfish Melanotaenia wilsoni sp. nov. based on 50 specimens, 19.3-33.3 mm SL. A combination of morphological characters is useful for separating the respective taxa with M. wilsoni sp. nov. the most distinctive, typically having fewer vertebrae, lateral scales, cheek scales, procurrent caudal rays and anal rays and proportionally a shorter maxilla and snout than either of the other two species. Useful characters for further separating M. sahulensis sp. nov. from M. maccullochi include slightly higher counts of vertebrae, lateral scales and anal rays and proportionally greater body depth, body width and pre-dorsal distance. Information on the known distribution, habitats and conservation status of the three species is summarised, with the Northern Territory species being a narrow-range endemic with specific environmental requirements.


Survival implications of cervical lymphadenectomy in patients with medullary thyroid cancer.
The relationship between extent of cervical lymphadenectomy along with the number of involved lymph nodes (LNs) removed and overall survival has not been well documented in patients with medullary thyroid carcinoma (MTC). This study investigates whether the overall number of LNs removed and the number of metastatic LNs are independent prognostic factors for overall survival. Data from patients with MTC in the Surveillance, Epidemiology, and End Results (SEER) registry database were examined. After categorizing the study population based on the number of overall LNs examined and the number of metastatic LNs, survival estimates were compared. The total number of examined LNs and their histopathological status were analyzed for their prognostic value in estimating overall survival. 593 patients were included in this study. Those with all negative LNs had the best overall survival; those with LNs examined and at least one positive LN had worst overall survival (p < 0.0001). The total number of examined LNs for both groups with negative and positive LNs was not associated with improved survival outcome (p = 0.41). In node-positive patients, each additional positive LN was significantly associated with an increase in overall mortality [hazard ratio (HR) = 1.05, 95% confidence interval (CI) = 1.02-1.08]. Cervical LN metastases conferred an independent risk for worse survival rate in MTC. Cervical lymphadenectomy is important for staging and regional disease control, however the extent of lymph node dissection, the overall number of lymph nodes removed along with removal of an increased number of involved lymph nodes do not confer a survival advantage. Future prospective studies are needed.


VHL induces renal cell differentiation and growth arrest through integration of cell-cell and cell-extracellular matrix signaling.
Mutations in the von Hippel-Lindau (VHL) gene are involved in the family cancer syndrome for which it is named and the development of sporadic renal cell cancer (RCC). Reintroduction of VHL into RCC cells lacking functional VHL [VHL(-)] can suppress their growth in nude mice, but not under standard tissue culture conditions. To examine the hypothesis that the tumor suppressor function of VHL requires signaling through contact with extracellular matrix (ECM), 786-O VHL(-) RCC cells and isogenic sublines stably expressing VHL gene products [VHL(+)] were grown on ECMs. Cell-cell and cell-ECM signalings were required to elicit VHL-dependent differences in growth and differentiation. VHL(+) cells differentiated into organized epithelial sheets, whereas VHL(-) cells were branched and disorganized. VHL(+) cells grown to high density on collagen I underwent growth arrest, whereas VHL(-) cells continued to proliferate. Integrin levels were up-regulated in VHL(-) cells, and cell adhesion was down-regulated in VHL(+) cells during growth at high cell density. Hepatocyte nuclear factor 1alpha, a transcription factor and global activator of proximal tubule-specific genes in the nephron, was markedly up-regulated in VHL(+) cells grown at high cell density. These data indicate that VHL can induce renal cell differentiation and mediate growth arrest through integration of cell-cell and cell-ECM signals.


Adolescent and nurse perspectives of psychotherapeutic interventions for PTSD delivered through task-shifting in a low resource setting.
This investigation compared the perceived effectiveness of supportive counselling (SC) and prolonged exposure for adolescents (PE-A) by treatment users (adolescents with PTSD) and non-specialist treatment providers (supervised nurses). Adolescent participants and nurse providers were purposively recruited to share their experiences of trial participation through face to face semi-structured in-depth interviews and treatment-specific focus groups (all recorded). Twelve adolescent participant transcripts (ten interviews and two focus groups) and three nurse provider transcripts were doubly transcribed. Thematic content analysis was applied using Atlas.ti software. Two emerging themes are presented in this paper: 1) Perceptions of the intervention and 2) Usefulness of the intervention. Regardless of treatment arm, adolescents experienced warm counselling relationships and described the process of extending trust to the counselor. Adolescents in the PE-A arm provided clear descriptions of session structure and treatment rationale compared with adolescents receiving SC. The most helpful tools were breathing retraining and imaginal exposure for PE-A and creation of distraction strategies during non-directive SC. Adolescents in both arms continued to use the techniques acquired during treatment and reported symptom improvement. Participants who received SC acknowledged ongoing reexperiencing. Nurses perceived SC to be an immediately transferable skill, but feedback on their preference for one intervention over the other was inconclusive. Both PTSD treatment strategies, implemented by non-specialists, were perceived as helpful. Overall, adolescents reported warm therapeutic relationships and a reduction in PTSD symptoms. Nurses stated that they would require institutional support to ensure delivery of these interventions in a scalable and sustainable manner.


Quantitation of glycosylated hemoglobin by boronate affinity chromatography.
Total hemoglobin glycosylation and the contribution of glycosylation at the N-terminus of the beta-chains and at "non"-beta-N-terminal positions were quantitated by use of boronate affinity and ion exchange chromatography. Glycohemoglobin (y) was found to correlate linearly (y = 1.92x + 0.53; r = 0.96) with HbA1c (x) and to contain approximately 50% beta-N-terminally glycosylated hemoglobin. This result is in agreement with the binding on boronate agarose of the various hemoglobin components resolved by cation exchange chromatography. An amount of glycohemoglobin similar to that of HbA1c was isolatable from HbA. A slope of less than 2 results because HbA1c is retained only to 93% and the intercept of the regression line reflects the partial adherence (65%) of HbA1a + b to the resin. These results confirm the occurrence of significant "non"-beta-N-terminal glycosylation and show that under optimal chromatographic conditions total glycohemoglobin can be determined with boronate affinity chromatography.


Vitamin A effects on UMR 106 osteosarcoma cells are not mediated by specific cytosolic receptors.
Retinol and retinoic acid at 20 microM altered cell morphology and inhibited cell proliferation of UMR 106 osteosarcoma cells in culture. No specific cytosolic binding proteins for retinol could be detected.


Newborn screening and cascade testing for FMR1 mutations.
We describe an ongoing pilot project in which newborn screening (NBS) for FMR1 mutations and subsequent cascade testing are performed by the MIND Institute at the University of California, Davis Medical Center (UCDMC). To date, out of 3,042 newborns initially screened, 44 extended family members have been screened by cascade testing of extended family members once a newborn is identified. Fourteen newborns (7 males and 7 females) and 27 extended family members (5 males and 22 females) have been identified with FMR1 mutations. Three family histories are discussed in detail, each demonstrating some benefits and risks of NBS and cascade testing for FMR1 mutations in extended family members. While we acknowledge inherent risks, we propose that with genetic counseling, clinical follow-up of identified individuals and cascade testing, NBS has significant benefits. Treatment for individuals in the extended family who would otherwise not have received treatment can be beneficial. In addition, knowledge of carrier status can lead to lifestyle changes and prophylactic interventions that are likely to reduce the risk of late onset neurological or psychiatric problems in carriers. Also with identification of carrier family members through NBS, reproductive choices become available to those who would not have known that they were at risk to have offspring with fragile X syndrome.


Hospitals' responses to nurse staffing shortages.
Hospitals have used a mix of short-term and long-term strategies to deal with nurse shortages, particularly efforts emphasizing nurse education, competitive compensation, and temporary staff. Interviews with health care leaders from Round Five of the Community Tracking Study indicate that these activities, in conjunction with other factors, have assisted in reducing shortages of hospital nurses. However, hospitals' actions have increased costs and raised concerns about their potential impact on patient care. Additionally, a large degree of doubt exists among hospitals about their ability to meet future nursing needs.


Predictors of survival following open and endovascular repair of abdominal aortic aneurysms.
Clinical decision making for asymptomatic abdominal aortic aneurysms (AAAs) weighs risk of aneurysm rupture, treatment hazards, and overall survival expectations. AAA diameter is the primary parameter in assessing rupture risk. Perioperative risk assessment has been extensively studied, and in-hospital mortality has been reduced to less than 8% with higher-risk open repair and less than 3% with endovascular repair. The purpose of this report is to determine risk factors that predict 2-year survival following open and endovascular AAA repair. We studied 334 patients enrolled in a multicenter clinical trial evaluating an endovascular graft in comparison to standard open repair of infrarenal AAA. Demographic, medical history, physical examination, laboratory, anatomic, procedural, and standardized risk score system variables were analyzed in a multivariable Cox proportional hazard model. Overall survival was 89% at 2 years. Heart disease, cancer, and stroke were the most common causes of death, and no deaths were due to AAA rupture. Cox modeling demonstrated that there were several independent predictors for death after AAA repair: smaller body mass index (p=0.005), Society for Vascular Surgery pulmonary risk score >or=1 (p=0.005), history of erectile dysfunction (p=0.008), history of heart valve replacement (p=0.008), lower preoperative platelet count (p=0.012), larger ratio of AAA diameter/proximal neck diameter (p=0.020), and lower ankle-brachial index (p=0.031). Age, gender, and open or endovascular treatment group are not significant independent risk factors for 2-year mortality in this study. Clinical, laboratory, and anatomic factors predict survival after open and endovascular repair of AAAs. With progressive reduction of in-hospital mortality, assessment of patient longevity after AAA repair has become a more important factor in clinical decision making. Use of valid predictors of patient survival will optimize resource utilization and improve overall patient outcomes. Better selection of patients for any method of repair may improve overall utility more than choice of open or endovascular techniques.


Assembly of nebulin into the sarcomeres of avian skeletal muscle.
In developing muscle, relatively little is known about the synthesis and incorporation of the large actin binding protein, nebulin, into the sarcomere. To determine the temporal pattern of nebulin assembly into the myofibrils of differentiating skeletal muscle cells, myofibril assembly was examined by immunofluorescence microscopy. The distribution of nebulin was compared to other myofibrillar and cytoskeletal proteins (myosin, titin, actin, desmin, tubulin). At the onset of differentiation, we observed that nebulin is first seen in a diffuse distribution throughout the cytoplasm. At this time, muscle specific myosin and titin are also distributed in this manner. Myosin and titin become associated with the nascent myofibrils prior to the addition of nebulin. The mature striated pattern of myosin and titin also preceded the development of striations with nebulin. After nebulin becomes organized into a striated pattern, actin filaments separate across the A-band and form thin filaments of uniform length. These patterns of assembly suggest that nebulin is required for restricting the lengths of the thin filaments. We have employed the strategy of using ethyl methane sulfonate and taxol to perturb myofibril assembly to examine interactions critical for the addition of nebulin to the developing sarcomeres. The same temporal pattern of assembly seen in the normal cultures was observed in the ethyl methane sulfonate treated cultures, but at a much slower rate. In cultures treated with the microtubule stabilizing drug taxol, the amount of stress fibers and nascent I-bands was greatly diminished as previously reported by others; however, nebulin was found associated with myofibrils in a mature striated distribution. In addition, our results indicate that the taxol treated cultures contain remnants of the Z-line. These results suggest that nebulin assembly into the myofibril requires interactions or anchorage at the Z-line and within the A-band.


Regulation of superoxide production in neutrophils: role of calcium influx.
Upon stimulation, activation of NADPH oxidase complexes in neutrophils produces a burst of superoxide anions contributing to oxidative stress and the development of inflammatory process. Store-operated calcium entry (SOCE), whereby the depletion of intracellular stores induces extracellular calcium influx, is known to be a crucial element of NADPH oxidase regulation. However, the mechanistic basis mediating SOCE is still only partially understood, as is the signal-coupling pathway leading to modulation of store-operated channels. This review emphasizes the role of calcium influx in the control of the NADPH oxidase and summarizes the current knowledge of pathways mediating this extracellular calcium entry in neutrophils. Such investigations into the cross-talk between NADPH oxidase and calcium might allow the identification of novel pharmacological targets with clinical use, particularly in inflammatory diseases.


Functional and genomic diversity of methylotrophic Rhodocyclaceae: description of Methyloversatilis discipulorum sp. nov.
Three strains of methylotrophic Rhodocyclaceae (FAM1(T), RZ18-153 and RZ94) isolated from Lake Washington sediment samples were characterized. Based on phylogenetic analysis of 16S rRNA gene sequences the strains should be assigned to the genus Methyloversatilis. Similarly to other members of the family, the strains show broad metabolic capabilities and are able to utilize a number of organic acids, alcohols and aromatic compounds in addition to methanol and methylamine. The main fatty acids were 16:1ω7c (49-59%) and 16:0 (32-29%). Genomes of all isolates were sequenced, assembled and annotated in collaboration with the DOE Joint Genome Institute (JGI). Genome comparison revealed that the strains FAM1T, RZ18-153 and RZ94 are closely related to each other and almost equally distant from two previously described species of the genus Methyloversatilis, Methyloversatilis universalis and Methyloversatilis thermotolerans. Like other methylotrophic species of the genus Methyloversatilis, all three strains possess one-subunit PQQ-dependent ethanol/methanol dehydrogenase (Mdh-2), the N-methylglutamate pathway and the serine cycle (isocitrate lyase/malate synthase, Icl/ms(+) variant). Like M. universalis, strains FAM1(T), RZ18-153 and RZ94 have a quinohemoprotein amine dehydrogenase, a tungsten-containing formaldehyde ferredoxin oxidoreductase, phenol hydroxylase, and the complete Calvin cycle. Similarly to M. thermotolerans, the three strains possess two-subunit methanol dehydrogenase (MxaFI), monoamine oxidase (MAO) and nitrogenase. Based on the phenotypic and genomic data, the strains FAM1(T), RZ18-153 and RZ94 represent a novel species of the genus Methyloversatilis, for which the name Methyloversatilis discipulorum sp. nov. is proposed. The type strain is FAM1(T) ( = JCM 30542(T) = VKM = B-2888(T)).


Functional analysis of the rat soft palate by real-time wireless electromyography.
The aim of this study was to analyze the function of the palatal muscles in vivo by real-time wireless electromyography in rats. The effects of palatal wounding were also analyzed. Microelectrodes were implanted six rats; in the masseter muscle (two-rats) for comparison, in the unwounded soft palate (two-rats) and the soft palate that received a surgical wound (two-rats). Two weeks after implantation, a wound was made in the soft palate using a 1 mm biopsy-punch. Electromyographic measurements and video-recordings were taken weekly to monitor train-duration and peak-amplitude during eating, grooming and drinking. The train-duration of the masseter muscle during eating was 0.49 ± 0.11 s (rat-1) and 0.56 ± 0.09 s (rat-2), which was higher than during grooming. In the unwounded soft palate the train-duration during eating was 0.63 ± 0.12 s (rat-1) and 0.69 ± 0.069 s (rat-2), which was higher than during grooming and drinking. The peak-amplitude for eating in the normal soft palate before surgery was 0.31 ± 0.001 mV (rat-1) and 0.33 ± 0.02 mV (rat-2). This decreased to 0.23 ± 0.03 mV and 0.25 ± 0.11 mV respectively, after surgery. For drinking the peak-amplitude was 0.30 ± 0.01 mV (rat-1) and 0.39 ± 0.01 mV (rat-2) before surgery, which decreased to 0.23 ± 0.09 mV and 0.20 ± 0.14 mV respectively, after surgery. The reduced peak-amplitude suggests impaired soft palate function after wounding. This is the first study into the in vivo function of the soft palate after surgical wounding. This model will contribute to develop strategies to improve soft palate function in patients.


Laparoscopic surgery for complex and recurrent Crohn's disease.
Crohn's disease (CD) is a chronic inflammatory disease of digestive tract. Approximately 70% of patients with CD require surgical intervention within 10 years of their initial diagnosis, despite advanced medical treatment alternatives including biologics, immune suppressive drugs and steroids. Refractory to medical treatment in CD patients is the common indication for surgery. Unfortunately, surgery cannot cure the disease. Minimally invasive treatment modalities can be suitable for CD patients due to the benign nature of the disease especially at the time of index surgery. However, laparoscopic management in fistulizing or recurrent disease is controversial. Intractable fibrotic strictures with obstruction, fistulas with abscess formation and hemorrhage are the surgical indications of recurrent CD, which are also complicating laparoscopic treatments. Nevertheless, laparoscopy can be performed in selected CD patients with safety, and may provide better outcomes compared to open surgery. The common complication after laparoscopic intervention is postoperative ileus seems and this may strongly relate excessive manipulation of the bowel during dissection. But additionally, unsuccessful laparoscopic attempts requiring conversion to open surgery have been a major concern due to presumed risk of worse outcomes. However, recent data show that conversions do not to worsen the outcomes of colorectal surgery in experienced hands. In conclusion, laparoscopic treatment modalities in recurrent CD patients have promising outcomes when it is used selectively.


Neurophysiological evidence for the interplay of speech segmentation and word-referent mapping during novel word learning.
Learning a new language requires the identification of word units from continuous speech (the speech segmentation problem) and mapping them onto conceptual representation (the word to world mapping problem). Recent behavioral studies have revealed that the statistical properties found within and across modalities can serve as cues for both processes. However, segmentation and mapping have been largely studied separately, and thus it remains unclear whether both processes can be accomplished at the same time and if they share common neurophysiological features. To address this question, we recorded EEG of 20 adult participants during both an audio alone speech segmentation task and an audiovisual word-to-picture association task. The participants were tested for both the implicit detection of online mismatches (structural auditory and visual semantic violations) as well as for the explicit recognition of words and word-to-picture associations. The ERP results from the learning phase revealed a delayed learning-related fronto-central negativity (FN400) in the audiovisual condition compared to the audio alone condition. Interestingly, while online structural auditory violations elicited clear MMN/N200 components in the audio alone condition, visual-semantic violations induced meaning-related N400 modulations in the audiovisual condition. The present results support the idea that speech segmentation and meaning mapping can take place in parallel and act in synergy to enhance novel word learning.


Infection dynamics of two renal myxozoans in hatchery reared fry and juvenile Atlantic cod Gadus morhua L.
In order to study the infection dynamics of 2 renal myxozoans, Zschokkella hildae Auerbach, 1910 and Gadimyxa atlantica Køie, Karlsbakk and Nylund, 2007 in cultured Atlantic cod, Gadus morhua L. aged 3-19 months, a specific single-round PCR assay and a double-label in situ hybridization protocol were developed. The results demonstrated that the 2 myxozoans show spatial separation of their development with regard to spore formation inside the renal tubules versus the collecting ducts and ureters, as well as temporal separation with Z. hildae proliferating and developing spores only once the G. atlantica infection decreases, despite the presence of both myxozoans in the smallest fry studied. These results strongly suggest within-host competition of the 2 myxozoans with potential suppression of Z. hildae by G. atlantica until G. morhua acquires immunity against G. atlantica. The quantification of the G. atlantica infection inside the renal tubules before and after a 29-day experimental growth performance study using fry from hatcheries with differing filtration systems showed that the intensity of infection with G. atlantica seems to be controlled if prolonged exposure to the myxozoan transmission stages takes place from hatching onwards. Surprisingly, growth rates in the trial were inversely affected suggesting that G. atlantica does not negatively influence cod fry growth performance.


Sequence lasing in a gain-switched Yb3+,Er(3+)-doped silica double-clad fiber laser.
Experimental results relating to the gain-switched operation of a double-clad Yb3+,Er(3+)-doped silica fiber laser that is pulse pumped with the output from a flash-lamp-pumped Ti:sapphire laser are presented. For all the configurations of the fiber laser that we studied, the 2F5/2-->2F7/2 laser transition of the Yb3+ ion lased prior to laser emission from the 4I13/2-->4I15/2 transition of the Er3+ ion. To the best of our knowledge, this is the first reported operation of sequence lasing in the Yb3+,Er(3+)-codoped system. This succession of laser pulses deduced from the measurements of this investigation is a consequence of both the short intense pump pulse and the short 900-nm wavelength of the pump that does not overlap with any important excited-state absorption transitions. We believe that the predominant interionic interaction during the course of the pump pulse is the double-energy transfer to the Er3+ ion acting twice from the 2F5/2 energy level of the Yb3+ donor ion. A maximum total output of 1.65 mJ is obtained (1.38 mJ from the 2F5/2-->2F7/2 transition of Yb3+ and 0.27 mJ from the 4I13/2-->4I15/2 transition of Er3+) from a nonoptimized configuration of the fiber laser. The wavelength of the output from the fiber laser was measured to vary approximately linearly with fiber length from 1040 to 1046 nm for the Yb(3+)-based laser and 1535 to 1541 nm for the Er(3+)-based laser.


A nonsuppressible increase of glucagon secretion by isolated islets of high-protein-fed rats.
We recently demonstrated increased plasma glucagon but nomal insulin in rats fed a high-protein, carbohydrate-free (HP) diet; however, other investigators have reported that both plasma glucagon and insulin are increased after protein feeding. For this reason, we have investigated the ffects of an HP diet on pancreatic secretion of insulin and glucagon. Male rats were fed an HP or control diet for one, three, or five days, and, at the end of the feeding period, blood was taken for glucose, insulin, and glucagon determinations. Additional animals fed the HP and control diets for up to 10 days were sacrificed, the pancreases removed, and islets of Langerhans isolated. Islets were incubated for 30 minutes in media with glucose concentration of1.7, 8.3, 16.7, or 33.4 mM. Insulin and glucagon secreted into the media were determined by radioimmunoassay. Plasma insulin was markedly reduced after one day of HP feeding but gradually returned to normal by the fifth day. Plasma glucagon was not altered on day 1 but was significantly increased after three days of HP feeding. The I/G molar ratio, which declined precipitously on day 1, increased thereafter but, as shown previously, remained at a level that promotes gluconeogenesis for up to 10 days. Insulin secretion by isolated islets of control and HP rats increased more than 10-fold as medium glucose was raised from 1.7 to 16.7 mM. There was no difference in insulin release by the two groups of islets. Glucagon secretion by HP islest at low medium glucose remained normal during the first five days; however, beginning on day 3 there was gradual loss of the suppressive effect of high medium glucose on glucagon secretion. After one week of HP feeding, glucagon secretion at low medium glucose was doubled and there was complete lack of suppression of the elevated hormone production by high medium glucose. The alterations of alpha-cell function induced by HP feeding are similar to those found in human and experimental diabetes.


Dietary Enrichment with 20% Fish Oil Decreases Mucus Production and the Inflammatory Response in Mice with Ovalbumin-Induced Allergic Lung Inflammation.
The prevalence of asthma has increased in recent decades, which may be related to higher dietary intake of (n-6) polyunsaturated fatty acids (PUFA) and lower intake of (n-3) PUFA, e.g., those contained in fish oil. The objective of this study was to determine if dietary PUFA enrichment decreases mucus production or the inflammatory response associated with ovalbumin (OVA)-induced allergic lung inflammation. Mice (n = 10/group) were fed control, 20% fish oil, or 20% corn oil enriched diets for a total of 12 weeks. At 8 and 10 weeks, mice were given an intraperitoneal injection of saline (10 control-fed mice) or OVA (30 remaining mice). Once at 10 weeks and on 3 consecutive days during week 12, mice were challenged by nebulizing with saline or OVA. Mice were euthanized 24 hours after the last challenge and blood was collected for plasma FA analysis. Bronchoalveolar lavage (BAL) fluid was collected to determine cell composition and Th2-type cytokine (IL-4, IL-13) concentrations. Periodic acid-Schiff (PAS) + mucus-producing cells and CD45+ inflammatory cell infiltrates in lung tissue were quantified using morphometric analysis. Relative abundance of mRNA for mucin (Muc4, Muc5ac, and Muc5b) and Th2-type cytokine (IL-4, IL-5, and IL-13) genes were compared with ß-actin by qPCR. Supplementation with either corn oil or fish oil effectively altered plasma FA profiles towards more (n-6) FA or (n-3) FA, respectively (P < 0.0001). Sensitization and challenge with OVA increased the proportion of neutrophils, lymphocytes, and eosinophils, and decreased the proportion of macrophages and concentrations of IL-13 in BAL fluid; increased the percentage of PAS+ mucus-producing cells and CD45+ inflammatory cell infiltrates in lung tissue; and increased gene expression of mucins (Muc4, Muc5ac, and Muc5b) and Th2-type cytokines (IL-5 and IL-13) in lung tissue of control-fed mice. Dietary PUFA reversed the increase in PAS+ mucus-producing cells (P = 0.003). In addition, dietary enrichment with fish oil attenuated the percentage of CD45+ inflammatory cell infiltrates in lung tissue, and increased Muc4 and Muc 5b gene expression compared with OVA-sensitized and challenged control mice. In conclusion, dietary enrichment with either (n-3) or (n-6) PUFA decreased mucus production in lung tissues of OVA-sensitized and challenged mice. More specifically, enrichment with dietary (n-3) PUFA decreased CD45+ inflammatory cell infiltrates, thus inducing potentially beneficial changes in lung tissue of OVA-sensitized and challenged mice.


Expression profiling and local adaptation of Boechera holboellii populations for water use efficiency across a naturally occurring water stress gradient.
We studied the physiological basis of local adaptation to drought in Boechera holboellii, a perennial relative of Arabidopsis thaliana, and used cDNA-AFLPs to identify candidate genes showing differential expression in these populations. We compared two populations of B. holboellii from contrasting water environments in a reciprocal transplant experiment, as well as in a laboratory dry-down experiment. We continuously measured the water content of soils using time domain reflectometery (TDR). We compared populations for their water use efficiency (WUE), root/shoot ratios (R:S) and leaf mass per unit area (LMA) in the field and in the laboratory, and identified candidate genes that (i) responded plastically to water stress and (ii) were differentially expressed between the two populations. Genotypes from the drier site had higher WUE, which was attributable to a large reduction in transpirational water loss. The xeric-adapted population also had increased investment in root biomass and greater leaf mass per unit area. Reciprocal transplants in the field had significantly greater survival in their native habitat. In total, 450 cDNA-AFLP fragments showed significant changes between drought and control treatments. Furthermore, some genes showed genotype (population)-specific patterns of up- or down-regulation in response to drought. Three hundred cDNA-AFLP bands were sequenced leading to the identification of cDNAs coding for proteins involved in signal transduction, transcriptional regulation, redox regulation, oxidative stress and pathways involved in stress adaptation. Some of these proteins could contribute a physiological advantage under drought, making them potential targets for natural selection.


The evolution of mutator genes in bacterial populations: the roles of environmental change and timing.
Recent studies have found high frequencies of bacteria with increased genomic rates of mutation in both clinical and laboratory populations. These observations may seem surprising in light of earlier experimental and theoretical studies. Mutator genes (genes that elevate the genomic mutation rate) are likely to induce deleterious mutations and thus suffer an indirect selective disadvantage; at the same time, bacteria carrying them can increase in frequency only by generating beneficial mutations at other loci. When clones carrying mutator genes are rare, however, these beneficial mutations are far more likely to arise in members of the much larger nonmutator population. How then can mutators become prevalent? To address this question, we develop a model of the population dynamics of bacteria confronted with ever-changing environments. Using analytical and simulation procedures, we explore the process by which initially rare mutator alleles can rise in frequency. We demonstrate that subsequent to a shift in environmental conditions, there will be relatively long periods of time during which the mutator subpopulation can produce a beneficial mutation before the ancestral subpopulations are eliminated. If the beneficial mutation arises early enough, the overall frequency of mutators will climb to a point higher than when the process began. The probability of producing a subsequent beneficial mutation will then also increase. In this manner, mutators can increase in frequency over successive selective sweeps. We discuss the implications and predictions of these theoretical results in relation to antibiotic resistance and the evolution of mutation rates.


The SOAR (Stroke subtype, Oxford Community Stroke Project classification, Age, prestroke modified Rankin) score strongly predicts early outcomes in acute stroke.
Previous prognostic scoring systems in predicting stroke mortality are complex, require multiple measures that vary with time and failed to produce a simple scoring system. The study aims to derive and internally validate a stroke prognostic scoring system to predict early mortality and hospital length of stay. Data from a U.K. multicenter stroke register were examined (1997-2010). Using a prior hypothesis based on our and others observations, we selected five patient-related factors (age, gender, stroke subtype, clinical classification, and prestroke disability) as candidate prognostic indicators. An 8-point score was derived based on multiple logistic regression model using four out of five variables. Performance of the model was assessed by plotting the estimated probability of in-hospital death against the actual probability by testing for overfitting (calibration) and area under the curve methods (discrimination). The total sample consisted of 12,355 acute stroke patients (ischemic stroke 91.0%). The score predicted both in-patient and seven-day mortality. The crude in-patient mortality were 1.57%, 4.02%, 10.65%, 21.41%, 46.60%, 62.72%, and 75.81% for those who scored 0, 1, 2, 3, 4, 5, and 6, respectively. The calibration of the model revealed no evidence of overfitting (estimated overfitting 0.001). The area under the curve values for both in-hospital and seven-day mortality were 0.79. The score predicted length of stay with a higher score was associated with longer median length of stay in those discharged alive and shorter median length of stay in those who died (P for both <0.001). A simple 8-point clinical score is highly predictive of acute stroke mortality and length of hospital stay. It could be used as prognostic tool in service planning and also to risk-stratify patients to use these outcomes as markers of stroke care quality across institutions.


Adolescent exercise in association with mortality from all causes, cardiovascular disease, and cancer among middle-aged and older Chinese women.
Little is known regarding the role of early-life exercise, a potentially modifiable factor, in long-term adult morbidity and mortality. We utilized the Shanghai Women's Health Study (SWHS) to investigate adolescent exercise in association with cancer, cardiovascular disease (CVD), and all-cause mortality among middle-aged and older women. The SWHS is a prospective cohort of 74,941 Chinese women ages 40 to 70 years recruited from 1996 to 2000. In-person interviews at enrollment assessed adolescent and adult exercise history, medical and reproductive history, and other lifestyle and socioeconomic (SES) factors. Mortality follow-up occurs via annual linkage to the Shanghai Vital Statistics Registry. Adjusted hazard ratios (HR) and 95% confidence intervals (CI) were derived from Cox regression models. Adjusting for birth year and other adolescent factors, adolescent exercise was associated with reduced risk of cancer, CVD, and total mortality [HRs (95% CI), 0.83 (0.72-0.95), 0.83 (0.70-0.98), and 0.78 (0.71-0.85), respectively for ≤1.33 hours (h)/week, and 0.83 (0.74-0.93), 0.62 (0.53-0.72), and 0.71 (0.66-0.77), respectively for >1.33 h/week (reference = none)]. Results were attenuated after adjustment for adult SES and lifestyle factors. Participation in sports teams was inversely associated with cancer mortality [HR (95% CI), 0.86 (0.76-0.97)]. Joint adolescent and adult exercise was associated with reduced risk of all-cause, CVD, and cancer mortality [HRs (95% CIs), 0.80 (0.72-0.89), 0.83 (0.69-1.00), and 0.87 (0.74-1.01), respectively], adjusting for adult/adolescent factors, and adolescence exercise only was inversely associated with cancer mortality [HR (95% CI), 0.84 (0.71-0.98)]. Adolescent exercise participation, independent of adult exercise, was associated with reduced risk of cancer, CVD, and all-cause mortality. Results support promotion of exercise in adolescence to reduce mortality in later life.


Over-expression of p53 nuclear oncoprotein in colorectal adenomas.
p53 is a nuclear phosphoprotein which controls normal cell growth. Normal p53 protein is undetectable by standard immunohistochemical staining and the over-expression found in neoplastic cells correlates with the presence of point mutations of evolutionary conserved regions of the p53 gene. We examined the expression of p53 protein in a series of 36 colorectal adenomas (13 tubular, 17 tubulovillous, 6 villous) showing different degrees of dysplasia (11 mild, 19 moderate, 6 severe), 11 moderately differentiated adenocarcinomas (6 Duke's A, 4 Duke's B, 1 Duke's C) and 5 metaplastic polyps using the polyclonal antibody CM1 which recognises p53 protein in conventionally fixed and processed histological material. We found that 15 out of 36 colorectal adenomas showed p53 immunoreactivity, although in 4 positive cases (26%) the staining was very focal (less than 0.1% positive cells). More than 80% of severely dysplastic adenomas showed strong p53 immunoreactivity and this over-expression was correlated with increased cell proliferative rate as detected by the proliferating-cell-nuclear-antigen (PCNA) staining. p53 nuclear staining was also seen in 8 out of 11 (65%) colorectal adenocarcinomas as previously shown. Our data suggest that the p53 gene mutation, with the subsequent over-expression of the protein, occurs in colorectal adenomas and may therefore be a fundamental genetic event underlying the dysplasia and loss of proliferative control that are characteristic of adenomas with malignant potential.


Induction of synthesis of mitochondrial steroidogenic enzymes of bovine adrenocortical cells by analogs of cyclic AMP.
The long term action of cyclic AMP analogs to stimulate the synthesis of cytochromes P-450scc, P-45011 beta, and adrenodoxin has been studied utilizing confluent monolayers of adult bovine adrenocortical cells maintained for periods of time up to 72 h in the absence or presence of dibutyryl cyclic AMP (1 mM), 8-bromo cyclic AMP (1 mM), or ACTH (adrenocorticotropin) (10(-6) M). The synthesis of these proteins was examined by radiolabeling cellular proteins with [35S]methionine or else by translating RNA extracted from such cells in a cell-free system in the presence of [35S]methionine. In each case, the protein under study was immunoprecipitated utilizing specific antisera, or IgG fractions prepared from such antisera. ACTH and both analogs of cyclic AMP caused an increase in the synthesis of cytochrome P-450scc which reached a maximum 36-48 h after addition, and then declined. On the other hand, butyric acid (1 mM) had no effect on the synthesis of cytochrome P-450scc. Cytochrome P-450scc activity measured as pregnenolone production by both intact cells or isolated mitochondria from such cells was increased following incubation of cells with either dibutyryl cyclic AMP or ACTH. The binding of rabbit anti-cytochrome P-450scc IgG was also increased in cells incubated with dibutyryl cyclic AMP or ACTH as estimated by immunofluorescence microscopy using fluorescein-tagged anti-rabbit IgG. Furthermore, dibutyryl cyclic AMP and ACTH both increased the synthesis of adrenodoxin and of cytochrome P-45011 beta, as well as the activity of 11 beta-hydroxylase. In addition, ACTH stimulated the secretion of cyclic AMP in a time- and concentration-dependent fashion. Thus, it is concluded that analogs of cyclic AMP can mimic the long term actions of ACTH to induce the synthesis of steroidogenic enzymes, and that this action of ACTH is likely mediated by cyclic AMP.