A spin crossover porous hybrid architecture for potential sensing applications.
Spin crossover cations have been successfully synthesized in the pores of a mesoporous robust Metal-Organic Framework (MOF) MIL-100(Al) through sequential introduction of Fe(iii) cations and a sal2trien ligand. The MIL-100(Al)@Fe(sal2trien) hybrid material retains its crystallinity and partial porosity compared to the parent MOF. The spin state of the Fe(sal2trien)+ cations can be modulated at room temperature through sorption of guest molecules, paving the way to the design of a new generation of sensors based on MOF@spin crossover complex solids.


Climate change and national crop wild relative conservation planning.
Climate change is likely to be one of the most important factors affecting our future food security. To mitigate negative impacts, we will require our crops to be more genetically diverse. Such diversity is available in crop wild relatives (CWRs), the wild taxa relatively closely related to crops and from which diverse traits can be transferred to the crop. Conservation of such genetic resources resides within the nation where they are found; therefore, national-level conservation recommendations are fundamental to global food security. We investigate the potential impact of climate change on CWR richness in Norway. The consequences of a 1.5 and 3.0 °C temperature rise were studied for the years 2030, 2050, 2070, 2080 and then compared to the present climate. The results indicate a pattern of shifting CWR richness from the south to the north, with increases in taxa turnover and in the numbers of threatened taxa. Recommendations for in situ and ex situ conservation actions over the short and long term for the priority CWRs in Norway are presented. The methods and recommendations developed here can be applied within other nations and at regional and global levels to improve the effectiveness of conservation actions and help ensure global food security.


Sleep duration and bone health measures in older men.
The associations between objective measures of sleep duration and bone outcomes in older men are unknown. No consistent, significant association was identified between sleep duration and bone mineral density (BMD) in the current analysis. However, future research should determine if vitamin D status modifies this relationship. Prior studies, predominantly in women, reported that long and short self-reported sleep duration are associated with lower BMD. Associations between actigraphy-determined sleep duration and BMD or bone turnover markers (BTMs) in older men are unknown. Men in The Osteoporotic Fractures in Men (MrOS) Study with wrist actigraphy and concurrent BMD assessment but without comorbidities affecting bone health were included. Sleep duration was considered as a continuous (N = 1926) and dichotomized variable where men were classified as getting the recommended (7-8 h/night; N = 478) or short (< 6 h/night; N = 577) sleep. The cross-sectional association between BMD, BTMs, and sleep duration was examined using a t test or linear regression, where appropriate, in unadjusted and adjusted models. There were no clinically or statistically significant differences in BMD at the L-spine, total hip, or femoral neck between men getting the recommended vs. short sleep duration, using actigraphy or self-reported sleep duration (all p ≥ 0.07). When sleep duration was considered as a continuous variable, femoral neck BMD was higher in men with longer self-reported sleep duration (β = 0.006 ±0.003, p = 0.02), but this was not significant after further adjustment. In men with low 25OHD (< 20 ng/mL), longer actigraphy-determined sleep duration was associated with higher total hip BMD (β = 0.016 ± 0.008; p = 0.04). Sleep duration and BTMs were not associated. Sleep duration was not associated with hip or L-spine BMD or BTMs in older men. Future research should determine if vitamin D status or other factors modify this relationship.


A novel software and conceptual design of the hardware platform for intensity modulated radiation therapy.
To deliver high quality intensity modulated radiotherapy (IMRT) using a novel generalized sparse orthogonal collimators (SOCs), the authors introduce a novel direct aperture optimization (DAO) approach based on discrete rectangular representation. A total of seven patients-two glioblastoma multiforme, three head & neck (including one with three prescription doses), and two lung-were included. 20 noncoplanar beams were selected using a column generation and pricing optimization method. The SOC is a generalized conventional orthogonal collimators with N leaves in each collimator bank, where N = 1, 2, or 4. SOC degenerates to conventional jaws when N = 1. For SOC-based IMRT, rectangular aperture optimization (RAO) was performed to optimize the fluence maps using rectangular representation, producing fluence maps that can be directly converted into a set of deliverable rectangular apertures. In order to optimize the dose distribution and minimize the number of apertures used, the overall objective was formulated to incorporate an L2 penalty reflecting the difference between the prescription and the projected doses, and an L1 sparsity regularization term to encourage a low number of nonzero rectangular basis coefficients. The optimization problem was solved using the Chambolle-Pock algorithm, a first-order primal-dual algorithm. Performance of RAO was compared to conventional two-step IMRT optimization including fluence map optimization and direct stratification for multileaf collimator (MLC) segmentation (DMS) using the same number of segments. For the RAO plans, segment travel time for SOC delivery was evaluated for the N = 1, N = 2, and N = 4 SOC designs to characterize the improvement in delivery efficiency as a function of N. Comparable PTV dose homogeneity and coverage were observed between the RAO and the DMS plans. The RAO plans were slightly superior to the DMS plans in sparing critical structures. On average, the maximum and mean critical organ doses were reduced by 1.94% and 1.44% of the prescription dose. The average number of delivery segments was 12.68 segments per beam for both the RAO and DMS plans. The N = 2 and N = 4 SOC designs were, on average, 1.56 and 1.80 times more efficient than the N = 1 SOC design to deliver. The mean aperture size produced by the RAO plans was 3.9 times larger than that of the DMS plans. The DAO and dose domain optimization approach enabled high quality IMRT plans using a low-complexity collimator setup. The dosimetric quality is comparable or slightly superior to conventional MLC-based IMRT plans using the same number of delivery segments. The SOC IMRT delivery efficiency can be significantly improved by increasing the leaf numbers, but the number is still significantly lower than the number of leaves in a typical MLC.


Stacking faults in Zr(Fe, Cr)2 Laves structured secondary phase particle in Zircaloy-4 alloy.
Stacking faults (SFs) in secondary phase particles (SPPs), which generally crystallize in the Laves phase in Zircaloy-4 (Zr-4) alloy, have been frequently observed by researchers. However, few investigations on the nano-scale structure of SFs have been carried out. In the present study, an SF containing C14 structured SPP, which located at grain boundaries (GBs) in the α-Zr matrix, was chosen to be investigated, for its particular substructure as well as location, aiming to reveal the nature of the SFs in the SPPs in Zr-4 alloy. It was indicated that the SFs in the C14 structured SPP actually existed in the local C36 structured Laves phase, for their similarities in crystallography. The C14 → C36 phase transformation, which was driven by synchroshearing among the (0001) basal planes, was the formation mechanism of the SFs in the SPPs. By analyzing the strained regions near the SPP, a model for understanding the driving force of the synchroshear was proposed: the interaction between SPP and GB resulted in the Zener pinning effect, leading to the shearing parallel to the (0001) basal planes of the C14 structured SPP, and the synchroshear was therefore activated.


A genome-wide association study of type 2 diabetes in Finns detects multiple susceptibility variants.
Identifying the genetic variants that increase the risk of type 2 diabetes (T2D) in humans has been a formidable challenge. Adopting a genome-wide association strategy, we genotyped 1161 Finnish T2D cases and 1174 Finnish normal glucose-tolerant (NGT) controls with >315,000 single-nucleotide polymorphisms (SNPs) and imputed genotypes for an additional >2 million autosomal SNPs. We carried out association analysis with these SNPs to identify genetic variants that predispose to T2D, compared our T2D association results with the results of two similar studies, and genotyped 80 SNPs in an additional 1215 Finnish T2D cases and 1258 Finnish NGT controls. We identify T2D-associated variants in an intergenic region of chromosome 11p12, contribute to the identification of T2D-associated variants near the genes IGF2BP2 and CDKAL1 and the region of CDKN2A and CDKN2B, and confirm that variants near TCF7L2, SLC30A8, HHEX, FTO, PPARG, and KCNJ11 are associated with T2D risk. This brings the number of T2D loci now confidently identified to at least 10.


Empirical Retrieval of Surface Melt Magnitude from Coupled MODIS Optical and Thermal Measurements over the Greenland Ice Sheet during the 2001 Ablation Season.
Accelerated ice flow near the equilibrium line of west-central Greenland Ice Sheet (GIS) has been attributed to an increase in infiltrated surface melt water as a response to climate warming. The assessment of surface melting events must be more than the detection of melt onset or extent. Retrieval of surface melt magnitude is necessary to improve understanding of ice sheet flow and surface melt coupling. In this paper, we report on a new technique to quantify the magnitude of surface melt. Cloud-free dates of June 10, July 5, 7, 9, and 11, 2001 Moderate Resolution Imaging Spectroradiometer (MODIS) daily reflectance Band 5 (1.230-1.250μm) and surface temperature images rescaled to 1km over western Greenland were used in the retrieval algorithm. An optical-thermal feature space partitioned as a function of melt magnitude was derived using a one-dimensional thermal snowmelt model (SNTHERM89). SNTHERM89 was forced by hourly meteorological data from the Greenland Climate Network (GC-Net) at reference sites spanning dry snow, percolation, and wet snow zones in the Jakobshavn drainage basin in western GIS. Melt magnitude or effective melt (E-melt) was derived for satellite composite periods covering May, June, and July displaying low fractions (0-1%) at elevations greater than 2500m and fractions at or greater than 15% at elevations lower than 1000m assessed for only the upper 5 cm of the snow surface. Validation of E-melt involved comparison of intensity to dry and wet zones determined from QSCAT backscatter. Higher intensities (> 8%) were distributed in wet snow zones, while lower intensities were grouped in dry zones at a first order accuracy of ~ ±2%.


Ethanol Extract of Peanut Sprout Lowers Blood Triglyceride Levels, Possibly Through a Pathway Involving SREBP-1c in Rats Fed a High-Fat Diet.
The hypothesis of this study was that peanut sprout extracts (PSE) could reduce fat accumulation through activating the transcription of SREBP-1c genes. Sprague-Dawley (SD) were randomly assigned into two groups and fed the following diet for 4 weeks; 10 normal fat (NF, 7 g of fat/100 g diet) and 30 high fat (HF, 20 g of fat/100 g diet). After 4 weeks, the HF group was divided into three groups; HF, HF with 15 mg of PSE/kg diet (HF+low PSE, 0.025% resveratrol), and HF with 30 mg of PSE/kg diet (HF+high PSE, 0.05% resveratrol) and fed for an additional 5 weeks. The HF+high PSE group had significantly lower weight gain than the HF group. Plasma triglyceride (TG) level and the hepatic total lipid level were significantly lower in the HF+high PSE group compared to the HF group. Fecal excretions of total lipids, cholesterol, and TG in the HF+high PSE group tended to be higher than in the HF group, but these differences were not significant. The mRNA expressions of fatty acid synthase, glucose-6-phosphate dehydrogenase, and sterol regulatory element binding protein-c (SREBP-1c) were significantly lower in the HF+high PSE group than in the HF group. The mRNA expressions of hydroxy-3-methylglutaryl coenzyme A reductase and acyl-CoA cholesterol acyltransferase were significantly lower in the HF+high PSE groups compared to the HF group. The mRNA expression of cholesterol 7α-hydroxylase1 was significantly higher than the HF group in both the HF+low PSE and HF+high PSE groups, with much greater increase observed in the HF+high PSE group. In conclusion, consumption of PSE was effective for improving blood lipid levels, possibly by suppressing the expression of SREBP-1c, in rats fed a high-fat diet.


Validation of self-reported and hospital-diagnosed atrial fibrillation: the HUNT study.
Self-reported atrial fibrillation (AF) and diagnoses from hospital registers are often used to identify persons with AF. The objective of this study was to validate self-reported AF and hospital discharge diagnoses of AF among participants in a population-based study. Among 50,805 persons who participated in the third survey of the HUNT Study (HUNT3), 16,247 participants from three municipalities were included. Individuals who reported cardiovascular disease, renal disease, or hypertension in the main questionnaire received a cardiovascular-specific questionnaire. An affirmative answer to a question on physician-diagnosed AF in this second questionnaire defined self-reported AF diagnoses in the study. In addition, AF diagnoses were retrieved from hospital and primary care (PC) registers. All AF diagnoses were verified by review of hospital and PC medical records. A total of 502 HUNT3 participants had a diagnosis of AF verified in hospital or PC records. Of these, 249 reported their AF diagnosis in the HUNT3 questionnaires and 370 had an AF diagnosis in hospital discharge registers before participation in HUNT3. The sensitivity of self-reported AF in HUNT3 was 49.6%, specificity 99.2%, positive predictive value (PPV) 66.2%, and negative predictive value (NPV) 98.4%. The sensitivity of a hospital discharge diagnosis of AF was 73.7%, specificity 99.7%, PPV 88.5%, and NPV 99.2%. Use of questionnaires alone to identify cases of AF has low sensitivity. Extraction of diagnoses from health care registers enhances the sensitivity substantially and should be applied when estimates of incidence and prevalence of AF are studied.


A whole genome scan of SNP data suggests a lack of abundant hard selective sweeps in the genome of the broad host range plant pathogenic fungus Sclerotinia sclerotiorum.
The pathogenic fungus Sclerotinia sclerotiorum infects over 600 species of plant. It is present in numerous environments throughout the world and causes significant damage to many agricultural crops. Fragmentation and lack of gene flow between populations may lead to population sub-structure. Within discrete recombining populations, positive selection may lead to a 'selective sweep'. This is characterised by an increase in frequency of a favourable allele leading to reduction in genotypic diversity in a localised genomic region due to the phenomenon of genetic hitchhiking. We aimed to assess whether isolates of S. sclerotiorum from around the world formed genotypic clusters associated with geographical origin and to determine whether signatures of population-specific positive selection could be detected. To do this, we sequenced the genomes of 25 isolates of S. sclerotiorum collected from four different continents-Australia, Africa (north and south), Europe and North America (Canada and the northen United States) and conducted SNP based analyses of population structure and selective sweeps. Among the 25 isolates, there was evidence for two major population clusters. One of these consisted of 11 isolates from Canada, the USA and France (population 1), and the other consisted of nine isolates from Australia and one from Morocco (population 2). The rest of the isolates were genotypic outliers. We found that there was evidence of outcrossing in these two populations based on linkage disequilibrium decay. However, only a single candidate selective sweep was observed, and it was present in population 2. This sweep was close to a Major Facilitator Superfamily transporter gene, and we speculate that this gene may have a role in nutrient uptake from the host. The low abundance of selective sweeps in the S. sclerotiorum genome contrasts the numerous examples in the genomes of other fungal pathogens. This may be a result of its slow rate of evolution and low effective recombination rate due to self-fertilisation and vegetative reproduction.


Molecular analysis of the autoantibody response in peptide-induced autoimmunity.
Immunization of nonautoimmune BALB/c mice with multimeric DWEYSVWLSN, a peptide mimotope of DNA, induces anti-DNA and other lupus-associated Abs. To further investigate the pathogenesis of the autoantibody response induced by peptide immunization, we generated hybridomas from peptide-immunized mice that bound peptide, dsDNA, cardiolipin, Sm/ribonucleoprotein (RNP), or some combination of these Ags. Analysis of 24 IgM Abs led to the identification of three groups of Abs: 1) Abs reactive with peptide alone, 2) anti-peptide Abs cross-reactive with one or more autoantigens, and 3) autoantibodies that do not bind to peptide. The gene families and particular VH-VL combinations used in those hybridomas binding DNA were similar to those used in the anti-DNA response in spontaneous murine lupus. Another similarity to the spontaneous anti-DNA response was the generation of arginines in the complementarity-determining region-3 of DNA-binding hybridomas. Interestingly, one Ab had the VH-VL combination present in the original R4A anti-DNA Ab used to select the DWEYSVWLSN peptide from a phage display library. Many of the heavy and light chains displayed evidence of somatic mutation, suggesting that they were made by Ag-activated B cells. Analysis of the Ab repertoire in peptide-induced autoimmunity may provide insights into the generation of anti-DNA Abs following exposure to foreign Ag. Furthermore, the recovery of an Ab with the heavy and light chain combination of the Ab originally used to isolate the immunizing peptide confirms the utility of phage display peptide libraries in generating true molecular mimics.


Information and communication technologies and gender in climate change and green economy: Situating women's opportunities and challenges in Zambian policies and strategies.
Zambia's 2012 report on the United Nations Conference on Sustainable Development (RIO +20) identifies existing opportunities on the country's transitioning to green economy. The RIO +20 conference of 2012 has resulted in new momentum in addressing problems of sustainable development. However, this article argues that there are practical challenges that require paying attention to, especially those involving women. The article addressed one key question: To what extent can women participate in the transitioning process to green economy in Zambia and what opportunities and challenges exists? The study used document analysis to answer the above question. National policy documents were reviewed to understand interventions on environmental management. Whilst going through the documents, the study used gender analysis frameworks (education, skills, roles in family and society, access to infrastructure) to bring out qualitative and quantitative information on women. Using suggested green economy interventions in the literature as benchmark, qualitative analysis was used to project possible participation of women in green economy activities and possible challenges to be faced. The study found that participation of women will be limited despite existing opportunities because of challenges of access to information and communication technology infrastructures, low educational levels and skills and financial constraints. As Zambia undergoes a transitioning process, these limitations should be addressed in planned green economy policies and interventions to maximise benefits.


Synthesis of single crystalline two-dimensional transition-metal phosphides via a salt-templating method.
Transition-metal phosphides (TMPs) are considered as promising non-noble electrochemical catalysts for hydrogen evolution reaction (HER). Their highly active sites are located on certain facets, and single crystalline two-dimensional (2D) structures enable them to expose the most active facets for HER. However, the synthesis of single crystalline 2D TMPs is still a challenge owing to their intrinsically non-layered structures. Herein, we demonstrate the synthesis of various single crystalline 2D TMPs (Co2P, MoP2, Ni12P5 and WP2) by a salt-templating method. The as-synthesized 2D Co2P exhibited efficient electrocatalytic ability for HER with an overpotential of 41 mV at 10 mA cm-2 and a Tafel slope of 35 mV dec-1 in 0.5 M H2SO4 solution. We expect that the synthesis of 2D TMPs reported here will open the way to expand the family of 2D materials for electrocatalysis and beyond.


Functionalised bicyclic tetramates derived from cysteine as antibacterial agents.
Routes to bicyclic tetramates derived from cysteine permitting ready incorporation of functionality at two different points around the periphery of a heterocyclic skeleton are reported. This has enabled the identification of systems active against Gram-positive bacteria, some of which show gyrase and RNA polymerase inhibitory activity. In particular, tetramates substituted with glycosyl side chains, chosen to impart polarity and aqueous solubility, show high antibacterial activity coupled with modest gyrase/polymerase activity in two cases. An analysis of physicochemical properties indicates that the antibacterially active tetramates generally occupy physicochemical space with MW of 300-600, clog D7.4 of -2.5 to 4 and rel. PSA of 11-22%. This work demonstrates that biologically active 3D libraries are readily available by manipulation of a tetramate skeleton.


Protein kinase C-dependent tyrosine phosphorylation of p130cas in differentiating neuroblastoma cells.
The cell signaling docking protein p130cas became tyrosine-phosphorylated in SH-SY5Y human neuroblastoma cells during induced differentiation with 12-O-tetradecanoylphorbol-13-acetate (TPA) and serum or a combination of basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I). The differentiating cells develop a neuronal phenotype with neurites and growth cones and sustained activation of protein kinase C (PKC) and pp60c-src. The TPA-induced p130cas phosphorylation increased within 5 min of stimulation and persisted for at least 4 days, whereas bFGF/IGF-I-induced p130cas phosphorylation was biphasic. However, the increase in tyrosine phosphorylation of p130cas was not restricted to differentiation inducing stimuli. The phosphorylation was blocked by the specific PKC inhibitor GF 109203X, and transient transfection with active PKC-epsilon induced p130cas tyrosine phosphorylation. pp60c-src, known to directly phosphorylate p130cas in other cell systems, was not activated after stimulation with TPA or bFGF/IGF-I for up to 30 min, and the initial p130cas phosphorylation was resistant to the Src family kinase inhibitor herbimycin A. However, in long term stimulated cells, herbimycin A blocked the induced phosphorylation of p130cas. Also, overexpression of src induced phosphorylation of p130cas. p130cas protein and phosphorylated p130cas were present in growth cones isolated from differentiated SH-SY5Y cells. Inhibition of PKC activity in differentiating cells with GF 109203X leads to a rapid retraction of growth cone filopodia, and p130cas phosphorylation decreased transiently (within minutes). Growth cones isolated from these cells were virtually devoid of phosphorylated p130cas. These data suggest a function for p130cas as a PKC downstream target in SH-SY5Y cells and possibly also in their growth cones.


Magnesium intake and risk of amyotrophic lateral sclerosis: results from five large cohort studies.
A low magnesium intake has been suggested to be associated with amyotrophic lateral sclerosis (ALS) in pathological and case-control studies, but prospective studies in humans are lacking. The relation between dietary intake of magnesium and ALS risk was explored in five large prospective cohort studies (the Nurses' Health Study, the Health Professionals Follow-up Study, the Cancer Prevention Study II Nutrition Cohort, the Multiethnic Cohort Study, and the National Institutes of Health - AARP Diet and Health Study), comprising over 1,050,000 males and females contributing 1093 cases of ALS during a mean of 15 years of follow-up. Cox proportional hazards models were used within each cohort, and cohort-specific estimates were subsequently pooled using a random-effects model. Results demonstrated that dietary magnesium intake was not associated with ALS risk, relative risk 1.07, 95% confidence interval 0.88 - 1.31 comparing the highest quintile of intake with the lowest. This finding does not support a protective effect of magnesium intake on ALS risk. Further analyses should explore magnesium intake in combination with heavy metal exposure and genetic variants affecting magnesium absorption.


A mathematical model demonstrating indirect and overall effects of lactation therapy targeting subclinical mastitis in dairy herds.
A deterministic state-transition model for mastitis transmission was developed to explore population level effects of antibiotic treatment regimens targeting chronic subclinical mastitis caused by major gram-positive pathogens in lactating dairy cows. Behavior and sensitivity of model outputs to changes in key parameters were explored. Outcomes included the size of the state variables describing proportions of infected quarters and basic and effective reproductive numbers. Treatment effects were estimated by calculating proportional reductions in state variables at equilibrium for populations implementing a treatment program relative to populations with no intervention. In general the relationships between parameters were complex and non-linear, although the model outputs were especially sensitive to changes in the value of the transmission rate parameter. Interaction between the parameters resulted in large variations in treatment effect estimates. Effect estimates calculated from model outputs showed a quadratic curve with a clear optimum at low, but not the lowest, transmission rates. These results indicated that overall positive population level effects of lactation therapy would be realized for herds that have successfully implemented practices that reduce the transmission rate of pathogens. A key finding is that in herds with high transmission rates, treatment of chronically infected quarters was predicted to have little impact on the proportion of infected quarters and no positive population level effect in reducing the force of infection and new infection rates. Results of this study suggest that field trials to evaluate efficacy of antimicrobial treatment should include estimates of indirect treatment effects.


Involvement of CRAF1, a relative of TRAF, in CD40 signaling.
CD40 is a receptor on the surface of B lymphocytes, the activation of which leads to B cell survival, growth, and differentiation. A yeast two-hybrid screen identified a gene, CRAF1, encoding a protein that interacts directly with the CD40 cytoplasmic tail through a region of similarity to the tumor necrosis factor-alpha (TNF-alpha) receptor-associated factors. Overexpression of a truncated CRAF1 gene inhibited CD40-mediated up-regulation of CD23. A region of CRAF1 was similar to the TNF-alpha receptor-associated factors TRAF1 and TRAF2 and so defined a shared TRAF-C domain that was necessary and sufficient for CD40 binding and homodimerization. The CRAF1 sequence also predicted a long amphipathic helix, a pattern of five zinc fingers, and a zinc ring finger. It is likely that other members of the TNF receptor superfamily use CRAF-related proteins in their signal transduction processes.


Clinicopathologic Significance of EGFR Mutation and HPV Infection in Sinonasal Squamous Cell Carcinoma.
Sinonasal squamous cell carcinoma (SNSCC) is sometimes associated with high-risk human papillomavirus (HR-HPV) infection and inverted sinonasal papilloma or oncocytic sinonasal papilloma. Frequent mutations of EGFR and KRAS are reported in inverted sinonasal papilloma-related sinonasal squamous cell carcinoma (ISP-SCC) and oncocytic sinonasal papilloma-related SNSCC, respectively. Here, we attempted to determine the prevalence and the prognostic significances of these alterations in SNSCC. We retrospectively collected 146 SNSCCs, including 14 ISP-SCCs, and comprehensively analyzed the HR-HPV infection by human papillomavirus (HPV)-RNA in situ hybridization, EGFR gene copy number gain (CNG) by chromogenic in situ hybridization, and gene mutations in EGFR and KRAS by Sanger sequencing. HR-HPV was detected in 11 cases (7.5%), whereas all 14 ISP-SCCs were negative. EGFR mutations were present in 21 (14.7%) of 143 SNSCCs, including 13/14 (92.9%) ISP-SCCs and 8/129 (6.2%) non-ISP-SCCs (P<0.0001). The majority of EGFR mutations were exon 20 insertions, with the remainder composed of deletions and single-nucleotide substitutions in exons 19 and 20. All of 142 SNSCCs harbored no KRAS mutation. EGFR CNG was detected in 41 (28.1%) of 146 SNSCCs; all of them were HPV negative and 3 had EGFR mutations. Collectively, EGFR mutation, EGFR CNG, and HR-HPV were essentially mutually exclusive, and each subgroup had distinct clinicopathologic features. The HPV-negative/EGFR-mutant group, the HPV-negative/EGFR CNG-positive group, and the triple-negative group had significantly worse prognoses than the HPV-positive group (P=0.0265, 0.0264, and 0.0394, respectively). In conclusion, EGFR mutation may play a pathogenetically important role in some populations of SNSCCs, especially ISP-SCCs. The molecular subclassification of SNSCCs may contribute to prognostic prediction and molecular-targeted precision medicine.


Effective Reduction of Acute Ethanol Withdrawal by the Tetracycline Derivative, Tigecycline, in Female and Male DBA/2J Mice.
Alcohol use disorder (AUD) is a spectrum disorder characterized by mild to severe symptoms, including potential withdrawal signs upon cessation of consumption. Approximately five hundred thousand patients with AUD undergo clinically relevant episodes of withdrawal annually (New Engl J Med, 2003, 348, 1786). Recent evidence indicates potential for drugs that alter neuroimmune pathways as new AUD therapies. We have previously shown the immunomodulatory drugs, minocycline and tigecycline, were effective in reducing ethanol (EtOH) consumption in both the 2-bottle choice and drinking-in-the-dark paradigms. Here, we test the hypothesis that tigecycline, a tetracycline derivative, will reduce the severity of EtOH withdrawal symptoms in a common acute model of alcohol withdrawal (AWD) using a single anesthetic dose of EtOH in seizure sensitive DBA/2J (DBA) mice. Naïve adult female and male DBA mice were given separate injections of 4 g/kg i.p. EtOH with vehicle or tigecycline (0, 20, 40, or 80 mg/kg i.p.). The 80 mg/kg dose was tested at 3 time points (0, 4, and 7 hours) post EtOH treatment. Handling-induced convulsions (HICs) were measured before and then over 12 hours following EtOH injection. HIC scores and areas under the curve were tabulated. In separate mice, blood EtOH concentrations (BECs) were measured at 2, 4, and 7 hours postinjection of 4 g/kg i.p. EtOH in mice treated with 0 and 80 mg/kg i.p. tigecycline. AWD symptom onset, peak magnitude, and overall HIC severity were reduced by tigecycline drug treatment compared to controls. Tigecycline treatment was effective regardless of timing throughout AWD, with earlier treatment showing greater efficacy. Tigecycline showed a dose-responsive reduction in acute AWD convulsions, with no sex differences in efficacy. Importantly, tigecycline did not affect BECs over a time course of elimination. Tigecycline effectively reduced AWD symptoms in DBA mice at all times and dosages tested, making it a promising lead compound for development of a novel pharmacotherapy for AWD. Further studies are needed to determine the mechanism of tigecycline action.


Mycosis fungoides in young patients: clinical characteristics and outcome.
Mycosis fungoides (MF) can begin as early as the first decade of life. Few studies have reviewed MF in younger patients and none has been large enough to assess prognosis and outcome. We reviewed the clinical characteristics, prognosis, factors related to disease progression, and therapy in patients with MF younger than 35 years of age. Fifty-eight patients were entered into this retrospective cohort analysis. Significantly fewer patients with MF who are younger than 35 years presented with erythroderma (T4) and more with limited patch/plaque (T1) disease than older patients. Duration of skin disease before diagnosis of MF did not vary between the two groups. The long-term survival of younger patients with MF is significantly decreased when compared with a race-, age-, and sex-matched control population (p < 0.001). Disease-specific survivals (DSS) of younger and older patients are similar, but young patients show a slight but significantly better overall DSS (p < 0.02). However, DSS comparison of generalized patch/plaque (T2) and tumor stage (T3) patients with MF showed no significant difference between young and old patients (p=0.47, p=0.59). Patient age was not a significant predictor of survival when controlled for T-stage. Sixteen of 58 young patients with MF have died, 13 because of MF (22%), compared with 138 of 500 older patients (28%) who died as a result of MF. All younger patients with MF who progressed had at least T2 disease at presentation. Fifty of 56 young patients with MF and T1-T3 disease were treated initially with total skin electron beam or topical nitrogen mustard. The response to therapy was similar in younger and older patients with MF. T1 disease is more common and T4 disease is unusual in young patients with MF compared with an older population of patients with MF. Young patients with T1 disease, all of whom were treated with either topical nitrogen mustard or total skin electron beam therapy, or both therapies, showed no disease progression. Overall, young patients with MF showed slightly better DSS, but this was because of differences in stage distribution.


Growing evidence for diabetes susceptibility genes from genome scan data.
Genome-wide scans for linkage have provided one of the dominant approaches adopted by researchers in their efforts to identify genes responsible for the inherited component of type 2 diabetes susceptibility. Around 20 genome scans have now been completed, in a wide variety of populations. Integration of data from these diverse scans has proven far from trivial, but the contours of genome-wide linkage topography are steadily emerging from the fog of data. Identification of the calpain-10 gene as the probable basis for the chromosome 2q linkage seen in Mexican Americans has provided validation of this positional cloning approach. This report provides an update on recent type 2 diabetes genome scan data, focusing on several chromosomal regions where the evidence for linkage has considerably strengthened in the past year. The current and future value of genome-wide linkage information in the search for type 2 diabetes susceptibility effects is also discussed.


Health-related quality of life in end-stage renal disease patients, using the Missoula-Vitas quality of life index: a multicenter study.
Assessment of Health-related Quality of Life in chronic hemodialysis patients (CHD) is a predictive indicator of the outcome of the disease, including mortality and hospitalization. Regular surveys of the quality of life (QoL) in CHD patients have been conducted worldwide, using various, internationally validated and standardized tools, including the Missoula-VITAS Quality of Life Index scale (MVQOLI). The aim of this study was to examine the reliability and validity of the Bulgarian version of the Missoula-VITAS Quality of Life Index-15 (B-MVQOLI-15) and QoL in CHD patients using this instrument. Our study was designed as multi-center cross-sectional. It incorporated 263 end-stage renal disease (ESRD) patients on CHD from across the country and applied the B-MVQOLI-15. Internal consistency and convergent validity of the index were assessed. Non-parametric methods were used to evaluate the impact of demographic factors on the different dimensions scores and on the total score. The relationship between the total QoL score, the total MVQOLI-15 score and dimensions scores were measured based on Spearman's rho Correlation Coefficient. The total MVQOLI-15 score in the study was 16.44, which is slightly above the middle of the index scale. The patients with higher education were less satisfied with the level of their symptom control compared to patients with lower education. However, high education patients seem to manage better with everyday life compared to those with low education. Men seem to feel more satisfied than women when fulfilling their daily activities (P=0.026). Retired patients and unemployed expressed more satisfaction, compared to the employed (P=0.021). Also, patients on dialysis for over 5 years had lower QoL scores (P=0.043). B-MVQOLI-15 is a reliable instrument to measure QoL in Bulgarian patients with CHD. The majority of CHD patients rate their QoL as "Fair". Four of all five dimensions positively affect the QoL of CHD patients. The most important dimensions positively affecting the QoL of CHD patients were: interpersonal relationships and transcendent factors. Future studies are necessary to assess the adequacy of the delivered dialysis, the level of medico-social care and the needs of ESRD patients treated with CHD in order to improve their QoL.


Discrete responses of erythrocytes, platelets, and von Willebrand factor to shear.
Despite decades of technological advancements in blood-contacting medical devices, complications related to shear flow-induced blood trauma are still frequently observed in clinic. Blood trauma includes haemolysis, platelet activation, and degradation of High Molecular Weight von Willebrand Factor (HMW vWF) multimers, all of which are dependent on the exposure time and magnitude of shear stress. Specifically, accumulating evidence supports that when blood is exposed to shear stresses above a certain threshold, blood trauma ensues; however, it remains unclear how various constituents of blood are affected by discrete shears experimentally. The aim of this study was to expose blood to discrete shear stresses and evaluate blood trauma indices that reflect red cell, platelet, and vWF structure. Citrated human whole blood (n = 6) was collected and its haematocrit was adjusted to 30 ± 2% by adding either phosphate buffered saline (PBS) or polyvinylpyrrolidone (PVP). Viscosity of whole blood was adjusted to 3.0, 12.5, 22.5 and 37.5 mPa·s to yield stresses of 3, 6, 9, 12, 50, 90 and 150 Pa in a custom-developed shearing system. Blood samples were exposed to shear for 0, 300, 600 and 900 s. Haemolysis was measured using spectrophotometry, platelet activation using flow cytometry, and HMW vWF multimer degradation was quantified with gel electrophoresis and immunoblotting. For tolerance to 300, 600 and 900 s of exposure time, the critical threshold of haemolysis was reached after blood was exposed to 90 Pa for 600 s (P < 0.05), platelet activation and HMW vWF multimer degradation were 50 Pa for 600 s and 12 Pa for 300 s respectively (P < 0.05). Our experimental results provide simultaneous comparison of blood trauma indices and thus also the relation between shear duration and magnitude required to induce damage to red cells, platelets, and vWF. Our results also demonstrate that near-physiological shear stress (<12 Pa) is needed in order to completely avoid any form of blood trauma. Therefore, there is an urgent need to design low shear-flow medical devices in order to avoid blood trauma in this blood-contacting medical device field.


Microfibrillar-associated protein 4 (MFAP4) genes in catfish play a novel role in innate immune responses.
The lectin pathway of the complement system is characterized by two groups of soluble pattern recognition molecules, mannose-binding lectins (MBLs) and ficolins. These molecules recognize and bind carbohydrates in pathogens and activate complement leading to opsonization, leukocyte activation, and direct pathogen killing. While MBLs have been reported in many fish species, ficolins do not appear to be present in the teleost lineage, despite their importance in invertebrate and higher vertebrate innate immunity. A protein with a similar fibrinogen-like domain, microfibrillar-associated protein 4, MFAP4, is present in fish, albeit with no described immune function. We examined whether MFAP4 genes in fish may potentially act as pathogen receptors in the absence of ficolin. We isolated and characterized five MFAP4 genes from channel catfish. Linkage mapping and phylogenetic analysis indicated that at least three of the catfish MFAP4 genes are tightly clustered on a single chromosome, suggesting that they may have arisen through tandem duplication. Divergent, duplicated families of MFAP4 genes are also present in other teleost species. Expression analysis of the catfish MFAP4 transcripts revealed unique patterns of homeostatic expression among the genes in gill, spleen, skin, liver, and muscle. Expression of the five MFAP4 transcripts showed significant changes in expression as soon as 4h after infection with either Edwardsiella ictaluri or Flavobacterium columnare with modulation of expression continuing up to 7 d following pathogen exposure. Several different tissues and gene-specific patterns were captured and transcript expression changes of >30-fold were observed over the course of the bacterial challenges. Our results suggest a novel role for MFAP4 in teleost immune responses.


Comparative analysis of the transcriptomes of Populus trichocarpa and Arabidopsis thaliana suggests extensive evolution of gene expression regulation in angiosperms.
Sequencing of the Populus trichocarpa genome creates an opportunity to describe the transcriptome of a woody perennial species and establish an atlas of gene expression. A comparison with the transcriptomes of other species can also define genes that are conserved or diverging in plant species. Here, the transcriptome in vegetative organs of the P. trichocarpa reference genotype Nisqually-1 was characterized. A comparison with Arabidopsis thaliana orthologs was used to distinguish gene functional categories that may be evolving differently in a woody perennial and an annual herbaceous species. A core set of genes expressed in common among vegetative organs was detected, as well as organ-specific genes. Statistical tests identified chromatin domains, where adjacent genes were expressed more frequently than expected by chance. Extensive divergence was detected in the expression patterns of A. thaliana and P. trichocarpa orthologs, but transcription of a small number of genes appeared to have remained conserved in the two species. Despite separation of lineages for over 100 million yr, these results suggest that selection has limited transcriptional divergence of genes associated with some essential functions in A. thaliana and P. trichocarpa. However, extensive remodeling of transcriptional networks indicates that expression regulation may be a key determinant of plant diversity.


Phospholipase C-beta 2 interacts with mitogen-activated protein kinase kinase 3.
Phospholipase C (PLC)-beta enzymes (isoenzymes beta 1-beta 4) are activated by G protein subunits, leading to the generation of intracellular messengers which mobilize calcium and activate protein kinase C. It has recently been recognized that these enzymes interact with and are regulated by proteins other than G proteins. Using the yeast two-hybrid technique to screen a leukocyte library we identified mitogen-activated protein kinase kinase 3 (MKK3) as a partner of PLC-beta 2. The interaction was confirmed by co-immunoprecipitation assays which indicated that MKK3 interacts with PLC-beta 2, but not with other PLC-betas. PLC-beta 2 interacted weakly with MKK6, which is related to MKK3, but not with the other MKK3 tested. The region of PLC-beta 2 involved in the interaction with MKK3 was mapped to the C-terminus of PLC-beta 2. p38MAPK also co-immunoprecipitated with PLC-beta 2. The data suggest that PLC-beta 2 serves an unappreciated role assembling components of the p38MAPK signaling module.


A biodegradable polyphosphoester-functionalized poly(disulfide) nanocarrier for reduction-triggered intracellular drug delivery.
Stimuli-responsive and biodegradable polymeric carriers are of great importance for safe delivery and efficient release of chemotherapeutic agents. In this work, given the unique advantages of poly(disulfide)s and biodegradable polyphosphoesters, we designed and constructed a reduction-sensitive amphiphilic triblock copolymer poly(ethyl ethylene phosphate)-b-poly(disulfide)-b-poly(ethyl ethylene phosphate) (PEEP-PDS-PEEP) by combining thiol-disulfide polycondensation and ring-opening polymerization (ROP). The thiol-disulfide polycondensation between 1,6-hexanedithiol and 2,2'-dithiodipyridine yielded the linear telechelic pyridyl disulfide-terminated poly(disulfide)s, followed by the treatment with 2-mercaptoethanol to quantitatively produce dihydroxyl-terminated poly(disulfide)s, which was used to initiate the ROP reaction of 2-ethoxy-2-oxo-1,3,2-dioxaphospholane, generating ABA-type amphiphilic triblock copolymers. The chemical structures of various polymers were thoroughly characterized and verified using nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, gel permeation chromatography (GPC) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectroscopy. The resultant amphiphilic PEEP-PDS-PEEP could self-assemble into spherical nanoparticles in aqueous solution as evidenced from dynamic light scattering (DLS) and transmission electron microscopy (TEM) analyses. Hydrophobic anti-tumor drug doxorubicin (DOX) was used to study the encapsulation capacity of nanoparticles, the drug loading content (DLC) and drug loading efficiency (DLE) values were determined to be 11.2% and 31.5%, respectively. In vitro release studies indicated that DOX was released much faster under reductive conditions compared to physiological conditions, confirming their reduction-responsive release behavior owing to the scission of the poly(disulfide) segment and subsequent disintegration of nanoparticles. The cellular uptake study using a live cell imaging system demonstrated that this DOX-loaded nanoparticle can be internalized into HeLa cells and release DOX over time. Methyl thiazolyl tetrazolium (MTT) assay revealed the favorable cytocompatibility of a bare triblock copolymer toward both L929 and HeLa cells, whereas the DOX-loaded copolymer nanoparticles exhibited the lower inhibitory ability against HeLa and HepG2 cell proliferation than free DOX. This finding presents a strategy for the construction of biocompatible and reduction-responsive polymeric drug carriers.


The walkshop approach to science and technology ethics.
In research and teaching on ethical aspects of emerging sciences and technologies, the structure of working environments, spaces and relationships play a significant role. Many of the routines and standard practices of academic life, however, do little to actively explore and experiment with these elements. They do even less to address the importance of contextual and embodied dimensions of thinking. To engage these dimensions, we have benefitted significantly from practices that take us out of seminar rooms, offices and laboratories as well as beyond traditional ways of working and interacting. We have called one such practice the 'walkshop'. Through walkshops, we have spent several days walking together with our colleagues and students in open outdoor spaces, keeping a sustained intellectual discussion on ethical aspects of science, technology and innovation while moving through these landscapes. For us, this has generated useful opportunities to escape established hierarchies, roles and patterns of thought and to rethink conceptual and philosophical issues from new perspectives, under new attitudes and with renewed energy. In this paper we wish to highlight the potential benefits of the walkshop approach by sharing some of our experiences and describing how we have prepared for and carried out these events. We share this information in the hope that we may encourage others to both experiment with the walkshop approach and exchange information on their own innovative processes for research and teaching in science and engineering ethics.


Alginate films as macromolecular imprinted matrices.
Macromolecularly imprinted polymers have been developed to mimic the non-covalent interactions driving molecular recognition in nature. The creation of an engineered antibody mimic would allow for the development of customizable films for biomolecular sensing. To demonstrate this principle, a cross-linked alginate film has been imprinted with bovine serum albumin (BSA) using aqueous biocompatible gelation methods. The imprinting efficiency of the synthesized films imprinted with BSA was determined and compared to the non-specific uptake of complementary proteins which were not imprinted in the alginate matrix. It was found that the recognition of the BSA using an alginate film was 6.4 mg/g polymer, which compares favorably to previously reported macromolecularly imprinted networks. The absorption of non-imprinted cationic proteins by the alginate matrix demonstrates that overcoming non-specific binding needs to be a focus of future work in order to successfully employ these materials towards biomolecular sensing within a physiological environment.


Severity of infantile nystagmus syndrome-like ocular motor phenotype is linked to the extent of the underlying optic nerve projection defect in zebrafish belladonna mutant.
Infantile nystagmus syndrome (INS), formerly known as congenital nystagmus, is an ocular motor disorder in humans characterized by spontaneous eye oscillations (SOs) and, in several cases, reversed optokinetic response (OKR). Its etiology and pathomechanism is largely unknown, but misrouting of the optic nerve has been observed in some patients. Likewise, optic nerve misrouting, a reversed OKR and SOs with INS-like waveforms are observed in zebrafish belladonna (bel) mutants. We aimed to investigate whether and how misrouting of the optic nerve correlates with the ocular motor behaviors in bel larvae. OKR and SOs were quantified and subsequently the optic nerve fibers were stained with fluorescent lipophilic dyes. Eye velocity during OKR was reduced in larvae with few misprojecting optic nerve fibers and reversed in larvae with a substantial fraction of misprojecting fibers. All larvae with reversed OKR also displayed SOs. A stronger reversed OKR correlated with more frequent SOs. Since we did not find a correlation between additional retinal defects and ocular motor behavior, we suggest that axon misrouting is in fact origin of INS in the zebrafish animal model. Depending on the ratio between misprojecting ipsilateral and correctly projecting contralateral fibers, the negative feedback loop normally regulating OKR can turn into a positive loop, resulting in an increase in retinal slip. Our data not only give new insights into the etiology of INS but may also be of interest for studies on how the brain deals with and adapts to conflicting inputs.


From DTCA-PD to patient information to health information: the complex politics and semantics of EU health policy.
Between 2001 and 2011 the pharmaceutical industry, supported by DG Enterprise, was engaged in an ongoing campaign to repeal/amend the European Union (EU) ban on direct-to-consumer advertising of prescription drugs (DTCA-PD). As it became increasingly clear that the ban would not be repealed, DTCA-PD supporters tried to shift the debate away from advertising and towards the provision of 'patient information' and the rights of patients to access such information. Meanwhile, a variety of national and European health organizations, supported by DG SANCO, sought to maintain the ban and oppose the industry-supported 'patient information' campaign. Instead, they promoted a concept of 'health information' that included all aspects of citizens' health, not just pharmaceuticals. This article aims to analyse the transition from DTCA-PD to patient information to health information and examine its implications for EU health policy as a complex policy space. The article examines the emergence and development of EU health policy and the evolution of the DTCA-PD debate through the lens of complexity theory. It analyses the nature of the semantic, political and policy transition and asks why it occurred, what it tells us about EU health policy and future EU health legislation and how it may be understood from a complexity perspective. The article concludes that the complexity framework is ideally suited for the field of public health and, in particular, the DTCA-PD debate. Having successfully shifted the policy-focus of the debate to patients' rights and health information, opponents of the legislation are likely to face their next battle in the realm of cyberspace, where regulatory issues change the nature of advertising.


A thematic content analysis of how marketers promote energy drinks on digital platforms to young Australians.
This study aimed to explore the nature and extent of, and level of user-engagement with, appealing strategies used by the food industry to promote energy drinks on digital platforms. Thematic content analysis was employed to code the textual and visual elements of the data that were extracted from the online media pages of nine energy drinks, including posts on Facebook, YouTube, Twitter, and content of websites and advergames. Four main themes were derived from the 624 textual and visual elements of digital marketing content of energy drink brands, including online social connectivity, desired social identity, enhancement of performance and enhancement of mood. Energy drinks were popular on digital platforms, as evidenced by the large volume of user-engagement (e.g. 'likes' and 'comments') especially on social networking sites. Energy drink brands appear to target young people, given that the marketing appeals are likely attuned with young people's desires or aspirations. Implications for public health: To counter the effects of digital marketing on young people's health, regulations are needed to safeguard adolescents and young adults, as well as younger children, and consideration should be given to including all forms of marketing communication platforms, including the internet.


Utilisation of diagnostic computerised tomography imaging and immediate clinical outcomes in older people with stroke before and after introduction of the National Service Framework for older people. A comparative study of hospital-based stroke registry data (1997-2003): Norfolk experience.
how the National Service Framework (NSF) for older people in England might be associated with changes in clinically relevant stroke outcome has not been investigated. We looked for changes in computerised tomography (CT) scan rate, inpatient case-fatality rate (CFR), length of acute hospital stay and discharge destination for older people with stroke, compared with their younger counterparts, for a period before, and after, the introduction of the NSF. two periods, 4 years before and 2 years after the publication of the NSF, were selected to compare the above outcomes between three age categories: < 65, 65-84 and > or = 85 years of age. Annual summary data for these periods were compared for the magnitude of changes in all age categories for all outcomes measured between pre- and post-NSF periods. n = 5,219. Utilisation of CT imaging had increased in all age groups post-NSF, with the most significant improvement in the oldest group. This change was associated with a greater proportion of people who had CT in this age group being discharged home in the post-NSF period. There was no change in the mortality from stroke in any age group during the study. Although the length of acute hospital stay increased, this was associated with a higher percentage of home discharges particularly in > 65-year olds, suggesting better clinical outcome in those who survived. in this single-centre analysis, the post-NSF period appeared to be associated with improvement in outcome in older people with stroke. Continual monitoring using stroke registry data may help to assess whether these effects are sustained in the longer term.


Melatonin treatment enhances therapeutic effects of exosomes against acute liver ischemia-reperfusion injury.
This study tests the hypothesis that combined melatonin and exogenic adipose mesenchymal stem cell (ADMSC)-derived exosome treatment offers superior protection against liver ischemia-reperfusion (LIR) injury compared to either alone. In vitro studies utilized a macrophage cell line (RAW) pretreated with lipopolysaccharide and hepatocytes pretreated with melatonin or exosomes before hypoxia treatment, while in vitro experiments involved analyses of liver specimens from male adult Sprague-Dawley rats (n = 50) equally categorized into sham controls (SC), LIR only, LIR-exosome (100 µg, 30 minute post-LIR), LIR-melatonin (20 mg/kg, 30 minute post-LIR and 50 mg/kg at 6 and 18 hours post-LIR), and LIR-exosome-melatonin groups. In vitro studies showed suppression of inflammation (MIF, MMP-9, IL-1β, TNF-α, COX-2) and oxidative stress (NOX-1, NOX-2, oxidized protein)/apoptosis (cleaved caspase 3 and PARP) by exosome and exosome/melatonin treatment, respectively (all P<0.001). In vivo data demonstrated lowest liver injury score and plasma AST concentrations in LIR-exosome-melatonin group compared with other groups (P<0.001). Besides, expressions of inflammatory markers at protein (ICAM-1, IL-1β, MMP-9, TNF-α, NF-κB, RANTES) and cellular (CD3+, CD4+, CD8+, CD161+, CD11+, CD14+, F4/80) levels, and protein expressions of apoptosis (cleaved caspase-3, PARP), oxidative stress (NOX-1, NOX-2), DNA damage (γ-H2AX) and mitochondrial damage (cytosolic cytochrome-C) markers displayed a pattern similar to that of liver injury score, whereas protein expression of anti-oxidants (HO-1, NQO-1) showed progressive increase from SC to the combined treatment group (all P<0.001). In conclusion, combined exosome-melatonin regimen was superior to either alone in protecting the liver against ischemia-reperfusion injury.


The activity of natural cytotoxic cells is augmented by interleukin 2 and interleukin 3.
Murine natural killer (NK) and natural cytotoxic (NC) cells showed different patterns of augmentation of lytic activity after preincubation for 24 h with either poly-IC, interleukin 2 (IL-2), or interleukin 3 (IL-3): (a) Poly-IC augmented only NK cells, with no effect on NC activity, as we have previously observed (4); (b) IL-2 augmented both NK and NC activity; and (c) IL-3 augmented only NC lysis, without affecting NK activity. In addition, both precursor and the augmented effector cells showed differences in expression of the Qa-5 surface marker: NK precursors and effectors are Qa 5+, whereas NC precursors and effector cells are Qa-5-.


Enhancement of osteoblastic differentiation in alginate gel beads with bioactive octacalcium phosphate particles.
The present study investigated whether alginate (Alg) hydrogel microbeads have a role in maintaining mouse bone marrow stromal ST-2 cells and release the cells after being stimulated by synthetic octacalcium phosphate (OCP), which is a mineral crystal capable of stimulating osteoblastic differentiation during a conversion process to hydroxyapatite (HA). The ST-2 cell suspension in the alginate solution, which contained various concentrations of OCP granules with diameters less than 53 μm, was extruded drop-wise into a stirred gelation solution containing BaCl2 using an encapsulator with nitrogen gas stream. The Alg-microbeads (Alg/OCP · ST-2 microbeads) that were generated, which had a diameter of approximately 400 μm, were incubated for up to 14 d and then assessed for osteoblastic differentiation. Alg-microbeads with cells were also incubated to identify the possible conversion from OCP to HA. Osteoblast differentiation markers in ST-2 cells, alkaline phosphatase (ALP) and collagen type I, were up-regulated in the presence of higher amounts of OCP. X-ray diffraction analysis and Fourier transform infrared spectroscopy confirmed that the OCP tended to convert to HA over time, suggesting that the OCP in Alg-microbeads interacts three-dimensionally with ST-2 cells and stimulates its osteoblastic differentiation. The release of ST-2 cells from the microbeads was also estimated. ST-2 cells were identified outside of the microbeads, although the cell number tended to decrease with increasing OCP. These results suggest that Alg/OCP microbeads could be used as a vehicle to activate osteoblastic cells and deliver them to sites where bone regeneration is needed.


Genetic and other factors determining mannose-binding lectin levels in American Indians: the Strong Heart Study.
Mannose-binding lectin (MBL) forms an integral part of the innate immune system. Persistent, subclinical infections and chronic inflammatory states are hypothesized to contribute to the pathogenesis of atherosclerosis. MBL gene (MBL2) variants with between 12 to 25% allele frequency in Caucasian and other populations, result in markedly reduced expression of functional protein. Prospective epidemiologic studies, including a nested, case-control study from the present population, have demonstrated the ability of MBL2 genotypes to predict complications of atherosclerosis,. The genetic control of MBL2 expression is complex and genetic background effects in specific populations are largely unknown. The Strong Heart Study is a longitudinal, cohort study of cardiovascular disease among American Indians. A subset of individuals genotyped for the above mentioned case-control study were selected for analysis of circulating MBL levels by double sandwich ELISA method. Mean MBL levels were compared between genotypic groups and multivariate regression was used to determine other independent factors influencing MBL2 expression. Our results confirm the effects of variant structural (B, C, and D) and promoter (H and Y) alleles that have been seen in other populations. In addition, MBL levels were found to be positively associated with male gender and hemoglobin A1c levels, but inversely related to triglyceride levels. Correlation was not found between MBL and other markers of inflammation. New data is presented concerning the effects of known genetic variants on MBL levels in an American Indian population, as well as the relationship of MBL2 expression to clinical and environmental factors, including inflammatory markers.


Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1.
Expression of vascular endothelial growth factor (VEGF) is induced in cells exposed to hypoxia or ischemia. Neovascularization stimulated by VEGF occurs in several important clinical contexts, including myocardial ischemia, retinal disease, and tumor growth. Hypoxia-inducible factor 1 (HIF-1) is a heterodimeric basic helix-loop-helix protein that activates transcription of the human erythropoietin gene in hypoxic cells. Here we demonstrate the involvement of HIF-1 in the activation of VEGF transcription. VEGF 5'-flanking sequences mediated transcriptional activation of reporter gene expression in hypoxic Hep3B cells. A 47-bp sequence located 985 to 939 bp 5' to the VEGF transcription initiation site mediated hypoxia-inducible reporter gene expression directed by a simian virus 40 promoter element that was otherwise minimally responsive to hypoxia. When reporters containing VEGF sequences, in the context of the native VEGF or heterologous simian virus 40 promoter, were cotransfected with expression vectors encoding HIF-1alpha and HIF-1beta (ARNT [aryl hydrocarbon receptor nuclear translocator]), reporter gene transcription was much greater in both hypoxic and nonhypoxic cells than in cells transfected with the reporter alone. A HIF-1 binding site was demonstrated in the 47-bp hypoxia response element, and a 3-bp substitution eliminated the ability of the element to bind HIF-1 and to activate transcription in response to hypoxia and/or recombinant HIF-1. Cotransfection of cells with an expression vector encoding a dominant negative form of HIF-1alpha inhibited the activation of reporter transcription in hypoxic cells in a dose-dependent manner. VEGF mRNA was not induced by hypoxia in mutant cells that do not express the HIF-1beta (ARNT) subunit. These findings implicate HIF-1 in the activation of VEGF transcription in hypoxic cells.


Tuning inter-dot tunnel coupling of an etched graphene double quantum dot by adjacent metal gates.
Graphene double quantum dots (DQDs) open to use charge or spin degrees of freedom for storing and manipulating quantum information in this new electronic material. However, impurities and edge disorders in etched graphene nano-structures hinder the ability to control the inter-dot tunnel coupling, tC, the most important property of the artificial molecule. Here we report measurements of tC in an all-metal-side-gated graphene DQD. We find that tC can be controlled continuously about a factor of four by employing a single gate. Furthermore, tC, can be changed monotonically about another factor of four as electrons are gate-pumped into the dot one by one. The results suggest that the strength of tunnel coupling in etched graphene DQDs can be varied in a rather broad range and in a controllable manner, which improves the outlook to use graphene as a base material for qubit applications.


Climate and land use changes will degrade the distribution of Rhododendrons in China.
Biodiversity loss and variation in species responses to climate and land use change have been found across broad taxonomic groups. However, whether species from the same taxonomic group with distinct geographical ranges will respond differently is poorly understood. The aim of this study is to predict the potential impacts of future climate and land use change on the distribution of narrow- and wide-ranging Rhododendron species, and estimate their relative contribution in China. We applied the presence-only ecological niche model MaxEnt to predict the distribution of 10 narrow-ranging and 10 wide-ranging Rhododendron species for the year 2070, using three general circulation models and three scenarios of climate and land use change. We measured the predicted distribution change of each species using change ratio, distance and direction of core range shifts, and niche overlap using Schoener's D. We found that the distribution areas of six narrow-ranging species would decrease, of which one species would go extinct. The remaining four narrow-ranging species would experience range expansion. Distribution of all the wide-ranging Rhododendron species would decrease. All Rhododendrons will shift to the northwest. We conclude that Rhododendron species generally will be negatively affected by the climatic and land use change expected in 2070 from the three scenarios evaluated in this study, but some narrow-ranging species may be positively influenced. Narrow-ranging Rhododendron species are more vulnerable compared to wide-ranging Rhododendron species. This study demonstrated that the effects of climate and land use change on alpine and subalpine plant species is species-specific, thereby strengthening our understanding of the impacts of climate and land use change on plant distribution.


A novel 65 kDa RNA-binding protein in squid presynaptic terminals.
A polyclonal antibody (C4), raised against the head domain of chicken myosin Va, reacted strongly towards a 65 kDa polypeptide (p65) on Western blots of extracts from squid optic lobes but did not recognize the heavy chain of squid myosin V. This peptide was not recognized by other myosin Va antibodies, nor by an antibody specific for squid myosin V. In an attempt to identify it, p65 was purified from optic lobes of Loligo plei by cationic exchange and reverse phase chromatography. Several peptide sequences were obtained by mass spectroscopy from p65 cut from sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gels. BLAST analysis and partial matching with expressed sequence tags (ESTs) from a Loligo pealei data bank indicated that p65 contains consensus signatures for the heterogeneous nuclear ribonucleoprotein (hnRNP) A/B family of RNA-binding proteins. Centrifugation of post mitochondrial extracts from optic lobes on sucrose gradients after treatment with RNase gave biochemical evidence that p65 associates with cytoplasmic RNP complexes in an RNA-dependent manner. Immunohistochemistry and immunofluorescence studies using the C4 antibody showed partial co-labeling with an antibody against squid synaptotagmin in bands within the outer plexiform layer of the optic lobes and at the presynaptic zone of the stellate ganglion. Also, punctate labeling by the C4 antibody was observed within isolated optic lobe synaptosomes. The data indicate that p65 is a novel RNA-binding protein located to the presynaptic terminal within squid neurons and may have a role in synaptic localization of RNA and its translation or processing.


Cross-sectional and prospective study of the effects of GH therapy on metabolic panel in children with GH deficiency.
Numerous studies have shown that GH, in addition to promoting linear growth, exerts a key role in many metabolic processes. However, there are only few studies aiming at evaluating the metabolic panel of children with GH deficiency (GHD). The aims of the study were: to verify the presence of metabolic alterations in GHD children in comparison with age-matched controls and to check the possible effects of two year GH therapy on the metabolic parameters in GHD. cross-sectional and prospective; one center experience We enrolled 32 pediatric GHD patients (group A) and 33 sex- and age-matched healthy controls (group B). Baseline serum assays (lipid, insulin, glucose) were performed in both groups. GHD children underwent replacement therapy with GH. The same assays were repeated after 12 and 24 months of GH treatment. No differences, in basal assays were found between the two groups. In group A, after initiation of GH, there was a significant increase of basal insulin and HOMA- insulin resistance (IR) index (p <0.001). In children with severe GHD (peak GH <3 ng / ml), after beginning of GH therapy a significant improvement in the lipid profile was found (p < 0.05). a) At the time of diagnosis GHD children had a metabolic picture that was not different from non- GHD group; b) in children with severe GHD, the metabolic profile showed a trend towards at improvement after the initiation of replacement therapy with GH, with beneficial effects in terms of total cholesterol, LDL cholesterol and cardiovascular risk indices; c) GHD patients with unfavorable metabolic profile (high BMI and hypercholestorolemia) need a monitoring of glucose metabolism by periodical evaluations of insulin and HOMA - IR.


A study of automated self-assessment in a primary care student health centre setting.
We evaluated the advice given by a prototype self-assessment triage system in a university student health centre. Students attending the health centre with a new problem used the automated self-assessment system prior to a face-to-face consultation with the general practitioner (GP). The system's rating of urgency was available to the GP, and following the consultation, the GP recorded their own rating of the urgency of the patient's presentation. Full data were available for 154 of the 207 consultations. Perfect agreement, where both the GP and the self-assessment system selected the same category of advice, occurred in 39% of consultations. The association between the GP assessment and the self-assessment rankings of urgency was low but significant (rho = 0.19, P = 0.016). The self-assessment system tended to be risk averse compared to the GP assessments, with advice for more urgent level of care seeking being recommended in 86 consultations (56%) and less urgent advice in only 8 (5%). This difference in assessment of urgency was significant (P < 0.001). The agreement between self-assessed and GP-assessed urgency was not associated with symptom site or socio-demographic characteristics of the user. Although the self-assessment system was more risk averse than the GPs, which resulted in a high proportion of patients being triaged as needing emergency or immediate care, the self-assessment system successfully identified a proportion of patients who were felt by the GP to have a self-limiting condition that did not need a consultation. In its prototype form, the self-assessment system was not a replacement for clinician assessment and further refinement is necessary.


Living in a paradox--women's experiences of body and life-world after meningioma surgery.
The aims of the study were to describe how persons who have undergone benign meningioma surgery experience their body and life-world and to illuminate the meaning of the illness, operation and recovery. Meningioma, which is often benign, is the most commonly diagnosed brain tumour. Studies have shown that persons with meningioma tend to be more psychologically affected than persons with other brain tumours. To our knowledge, no previous studies have focused on the life-world or body experience of these persons. Narrative interviews with seven women conducted over a 6-month period (2008-2009) were analyzed using a phenomenological hermeneutical method. Findings. Four themes were identified: In between double threats: the tumour and operation: the participants seemed to perceive a paradox in the sense that the tumour was both benign and threatening and the surgery both curing and risky; the wound as an open gate: the wound is visibly healed, but perceived as unhealed; the suffering body overflows life: the participants were prevented from regaining their body, feeling fragile and oversensitive to impressions; and the need to be embraced by attention: taking the participants' needs seriously facilitated understanding of the diagnosis. The participants in this study described emotional distress when recovering from meningioma surgery. It is essential for nurses to be aware of the various meanings people ascribe to events that disrupt their lives and the transition process in connection with meningioma treatment.


Clinical characteristics and pharmacokinetics of purified soy isoflavones: single-dose administration to healthy men.
Soy isoflavones are potential cancer chemoprevention treatments. We conducted safety studies of purified unconjugated genistein, daidzein, and glycitein, and defined pharmacokinetic parameters for their absorption and metabolism. Thirty healthy men ingested a single dose of 1 of 2 isoflavone preparations purified from soy. The delivered doses of genistein (1, 2, 4, 8, or 16 mg/kg body wt) were higher than those previously administered to humans. Formulation A was composed of 90 +/- 5% genistein, 10% daidzein, and 1% glycitein. Formulation B was composed of 43% genistein, 21% daidzein, and 2% glycitein. We observed no clinically significant behavioral or physical changes after treatment. We observed elevations in lipoprotein lipase and hypophosphatemia that were possibly related to the treatment but that were associated with no clinical toxicity. Considerable quantities of isoflavones were excreted in urine as conjugates. The terminal elimination rate, elimination half-life, area under the curve, maximum plasma concentration, apparent systemic clearance, and volume of distribution were estimated for genistein and daidzein. The mean elimination half-lives with both formulations were 3.2 h for free genistein and 4.2 h for free daidzein. The mean pseudo half-lives were 9.2 h for total genistein and 8.2 h for total daidzein. Dietary supplements of purified unconjugated isoflavones administered to humans in single doses exceeding normal dietary intake manyfold resulted in minimal clinical toxicity. Genistein and daidzein (free and total) were rapidly cleared from plasma and excreted in urine.


Human granulosa cells are a site of sulphatase activity and are able to utilize dehydroepiandrosterone sulphate as a precursor for oestradiol production.
Dehydroepiandrosterone sulphate (DHEAS) is the most abundant androgen in the circulation and in ovarian follicular fluid. A steroid sulphatase accepting DHEAS as a substrate has been identified in the follicle, but the cellular location has not been determined. As DHEAS is also a potential source of oestrogen for endocrine-dependent tumours, a potent steroid sulphatase inhibitor oestrone-3-O-sulphamate (EMATE) has been developed which inhibits this activity in rat liver and mammary tumour. The aim of this study was to investigate human granulosa cells as a site of steroid sulphatase activity, to determine whether DHEAS can be utilized as a precursor for oestrogen synthesis and to investigate the inhibitory capacity of EMATE in these cells. Conversion of DHEAS to DHEA was assessed in luteinized granulosa cells by tritiated steroid assay following incubation with or without LH or insulin and steroid accumulation in the medium measured by RIA. The effects of EMATE were assessed by addition of a range of doses during the measurement of conversion of DHEAS to DHEA. Cells from three sizes of small follicles from an unstimulated ovary were also assessed for their ability to produce oestradiol from DHEAS. Sulphatase enzyme activity was present in all cells; the mean conversion of tritiated DHEAS to DHEA was 50% (range 4-65%). LH and EMATE inhibited and insulin stimulated this activity. Addition of DHEAS to granulosa cells caused a dose-dependent increase in oestradiol and androstenedione production with no change in progesterone concentration. LH increased the accumulation of oestradiol in the medium. DHEAS also stimulated oestradiol production by granulosa cells from small follicles. This is the first demonstration that granulosa cells are a site of sulphatase activity and that DHEAS can be utilized as a substrate for androstenedione and oestrogen production. This may be of physiological importance for both normal folliculogenesis and oestrogen-dependent tumour growth.


Driving forces for the self-assembly of graphene oxide on organic monolayers.
Graphene oxide (GO) flakes were self-assembled from solution on surfaces of self-assembled monolayers (SAMs), varying in the chemical structure of their head groups. The coverage density of GO relates to strength of attractive interaction, which is largest for Coulomb interaction provided by positively charged SAM head groups and negatively charged GO. A rough surface enhances the coverage density but with the same trend in driving force dependency. The self-assembly approach was used to fabricate field-effect transistors with reduced GO (rGO) as active layer. The SAMs as attractive layer for self-assembly remain almost unaffected by the reduction from GO to rGO and serve as ultra-thin gate dielectrics in devices, which operate at low voltages of maximum 3 V and exhibit a shift of the Dirac voltage related to the dipole moment of the SAMs.


Evaluation of Ochratoxin Recognition by Peptides Using Explicit Solvent Molecular Dynamics.
Biosensing platforms based on peptide recognition provide a cost-effective and stable alternative to antibody-based capture and discrimination of ochratoxin-A (OTA) vs. ochratoxin-B (OTB) in monitoring bioassays. Attempts to engineer peptides with improved recognition efficacy require thorough structural and thermodynamic characterization of the binding-competent conformations. Classical molecular dynamics (MD) approaches alone do not provide a thorough assessment of a peptide's recognition efficacy. In this study, in-solution binding properties of four different peptides, a hexamer (SNLHPK), an octamer (CSIVEDGK), NFO4 (VYMNRKYYKCCK), and a 13-mer (GPAGIDGPAGIRC), which were previously generated for OTA-specific recognition, were evaluated using an advanced MD simulation approach involving accelerated configurational search and predictive modeling. Peptide configurations relevant to ochratoxin binding were initially generated using biased exchange metadynamics and the dynamic properties associated with the in-solution peptide-ochratoxin binding were derived from Markov State Models. Among the various peptides, NFO4 shows superior in-solution OTA sensing and also shows superior selectivity for OTA vs. OTB due to the lower penalty associated with solvating its bound complex. Advanced MD approaches provide structural and energetic insights critical to the hapten-specific recognition to aid the engineering of peptides with better sensing efficacies.


Diosgenin induces hypoxia-inducible factor-1 activation and angiogenesis through estrogen receptor-related phosphatidylinositol 3-kinase/Akt and p38 mitogen-activated protein kinase pathways in osteoblasts.
Diosgenin, extracted from the root of wild yam (Dioscorea villosa), has been reported to demonstrate an opportunity for medical application. Vascular endothelial growth factor-A (VEGF-A) plays an important role in bone-related angiogenesis, a critical process occurring during bone formation and fracture healing. In this study, we examine whether diosgenin is able to induce VEGF-A expression and to promote angiogenesis in osteoblasts. For murine MC3T3-E1 preosteoblast-like cells, VEGF-A mRNA and protein expression seemed to be significantly elevated in response to diosgenin in a concentration-dependent fashion. Conditioned media prepared from cells treated with diosgenin induced strong angiogenic activity in either in vitro or ex vivo angiogenesis assay. Furthermore, diosgenin treatment increased the stability and activity of HIF-1alpha protein. Inhibition of HIF-1alpha activity by transfection with DN-HIF-1alpha significantly diminished diosgenin-mediated VEGF-A up-regulation. The use of pharmacological inhibitors or genetic inhibition revealed that both the phosphatidylinositol 3-kinase (PI3K)/Akt and p38 signaling pathways were potentially required for diosgenin-induced HIF-1 activation and subsequent VEGF-A up-regulation. It is noteworthy that an estrogen receptor binding assay revealed that diosgenin has the strong ability to replace [(3)H]estradiol bound to estrogen receptor (IC(50), 10 nM). In addition, the specific estrogen receptor antagonists ICI 182,780 (faslodex) and tamoxifen were noted to be able to strongly inhibit diosgenin-induced, src kinase-dependent Akt and p38 MAPK activation. Taken together, such results provide evidence that diosgenin up-regulates VEGF-A and promotes angiogenesis in preosteoblast-like cells by a hypoxia-inducible factor-1alpha-dependent mechanism involving the activation of src kinase, p38 MAPK, and Akt signaling pathways via estrogen receptor.


A rapid, computational approach for assessing interfraction esophageal motion for use in stereotactic body radiation therapy planning.
We present a rapid computational method for quantifying interfraction motion of the esophagus in patients undergoing stereotactic body radiation therapy on a magnetic resonance (MR) guided radiation therapy system. Patients who underwent stereotactic body radiation therapy had simulation computed tomography (CT) and on-treatment MR scans performed. The esophagus was contoured on each scan. CT contours were transferred to MR volumes via rigid registration. Digital Imaging and Communications in Medicine files containing contour points were exported to MATLAB. In-plane CT and MR contour points were spline interpolated, yielding boundaries with centroid positions, CCT and CMR. MR contour points lying outside of the CT contour were extracted. For each such point, BMR(j), a segment from CCT intersecting BMR(j), was produced; its intersection with the CT contour, BCT(i), was calculated. The length of the segment Sij, between BCT(i) and BMR(j), was found. The orientation θ was calculated from Sij vector components:θ = arctan[(Sij)y / (Sij)x]A set of segments {Sij} was produced for each slice and binned by quadrant with 0° < θ ≤ 90°, 90° < θ ≤ 180°, 180° < θ ≤ 270°, and 270° < θ ≤ 360° for the left anterior, right anterior, right posterior, and left posterior quadrants, respectively. Slices were binned into upper, middle, and lower esophageal (LE) segments. Seven patients, each having 3 MR scans, were evaluated, yielding 1629 axial slices and 84,716 measurements. The LE segment exhibited the greatest magnitude of motion. The mean LE measurements in the left anterior, left posterior, right anterior, and right posterior were 5.2 ± 0.07 mm, 6.0 ± 0.09 mm, 4.8 ± 0.08 mm, and 5.1 ± 0.08 mm, respectively. There was considerable interpatient variability. The LE segment exhibited the greatest magnitude of mobility compared with the middle and upper esophageal segments. A novel computational method enables personalized, nonuniform esophageal margins to be tailored to individual patients.


A closer look at the role of healthcare in the recent mortality decline in the Netherlands: results of a record linkage study.
Since 2002, Dutch mortality rates decreased rapidly after decades of stagnation. On the basis of indirect evidence, previous research has suggested that this decline was due to a sudden expansion of healthcare. We tested two corollaries of this hypothesis--first, that the decline was concentrated among those with ill-health and second, that the decline can be statistically accounted for by increases in healthcare utilisation. We linked the Dutch health interview survey to the mortality register and constructed two cohorts, consisting of 7691 persons interviewed in 2001/2002 and 8362 persons interviewed in 2007/2008, each with a 5-year mortality follow-up (659 deaths in total). The change in mortality between both cohorts was computed using Cox proportional hazard models. We estimated the change in mortality by severity of chronic conditions and with respect to the inclusion of indicators of healthcare utilisation. Between the two study cohorts, mortality declined by 15% (95% CI 2% to 29%), and mortality reduction was greatest for those suffering from fatal and non-fatal conditions with a decline of 58% (95% CI 35% to 78%). Even after adjustment for health status and risk factors, most indicators of healthcare utilisation were associated with higher instead of lower mortality and changes in healthcare utilisation did not explain the decline in mortality. Our results only partly confirm the hypothesis that an expansion of healthcare explains the recent mortality decline in the Netherlands. Owing to confounding by health status, it is difficult to reproduce the mortality-lowering effects of healthcare utilisation of individual level studies in the open population.


Intrinsic activity temporal structure reactivity to behavioural state change is correlated with depressive symptoms.
The brain's intrinsic activity plays a fundamental role in its function. In normal conditions this activity is responsive to behavioural context, changing as an individual switches between directed tasks and task-free conditions. A key feature of such changes is the movement of the brain between corresponding critical and sub-critical states, with these dynamics supporting efficient cognitive processing. Breakdowns in processing efficiency can occur, however, in brain disorders such as depression. It was therefore hypothesised that depressive symptoms would be related to reduced intrinsic activity responsiveness to changes in behavioural state. This was tested in a mixed group of major depressive disorder patients (n = 26) and healthy participants (n = 37) by measuring intrinsic EEG activity temporal structure, quantified with detrended fluctuation analysis (DFA), in eyes-closed (EC) and eyes-open task-free states and contrasting between the conditions. The degree to which DFA values changed between the states was found to correlate negatively with depressive symptoms. DFA values did not differ between states in those with higher symptom levels, meaning that the brain remained in a less flexible sub-critical condition. This sub-critical condition in the EC state was further found to correlate with levels of maladaptive rumination. This may reflect a general cognitive inflexibility resulting from a lack in neural activity reactivity that may predispose people to overly engage in self-directed attention. These results provide an initial link between intrinsic activity reactivity and psychological features found in psychiatric disorders.


Neutrophils sense flow-generated stress and direct their migration through alphaVbeta3-integrin.
During inflammation neutrophils are recruited from the blood onto the surface of microvascular endothelial cells. In this milieu the presence of soluble chemotactic gradients is disallowed by blood flow. However, directional cues are still required for neutrophils to migrate to the junctions of endothelial cells where extravasation occurs. Shear forces generated by flowing blood provide a potential alternative guide. In our flow-based adhesion assay neutrophils preferentially migrated in the direction of flow when activated after attachment to platelet monolayers. Neutralizing alphaVbeta3-integrin with monoclonal antibodies or turning the flow off randomized the direction of migration without affecting migration velocity. Purified, immobilized alphaVbeta3-integrin ligands, CD31 and fibronectin, could both support flow-directed neutrophil migration in a concentration-dependent manner. Migration could be randomized by neutralizing alphaVbeta3-integrin interactions with the substrate using antibodies or Arg-Gly-Asp-containing peptide. These results exemplify mechanical signal transduction through integrin-ligand interactions and reveal a guidance system that was hitherto unknown in neutrophils. In more general terms, it demonstrates that cells can use integrin molecules to "sample" their physical microenvironment through adhesion and use this information to modulate their behavior.


Potential role of exosome-based allorecognition pathways involved in lung transplant rejection.
Innate and adaptive immunity both contribute to allorecognition mechanisms that drive rejection after lung transplantation. Classic allorecognition pathways have been extensively described, but there continues to be several unanswered questions. Exosome research appears to be a novel and potentially significant area of allorecognition research and could be the missing link that answers some existing questions. This article reviews literature that is associated with allorecognition pathways and the role of exosomes in alloreactivity.


Educational differences in mobility: the contribution of physical workload, obesity, smoking and chronic conditions.
In earlier studies, determinants of socioeconomic gradient in mobility have not been measured comprehensively. To assess the contribution of chronic morbidity, obesity, smoking and physical workload to inequalities in mobility. This was a cross-sectional study on 2572 persons (76% of a nationally representative sample of the Finnish population aged > or = 55 years). Mobility limitations were measured by self-reports and performance rates. According to a wide array of self-reported and test-based indicators, persons with a lower level of education showed more mobility limitations than those with a higher level. The age-adjusted ORs for limitations in stair climbing were threefold in the lowest-educational category compared with the highest one (OR 3.3 in men and 2.9 in women for self-reported limitations, and 3.5 in men and 2.2 in women for test-based limitations). When obesity, smoking, work-related physical loading and clinically diagnosed chronic diseases were simultaneously accounted for, the educational differences in stair-climbing limitations vanished or were greatly diminished. In women, obesity contributed most to the differences, followed by a history of physically strenuous work, knee and hip osteoarthritis and cardiovascular diseases. In men, diabetes, work-related physical loading, musculoskeletal diseases, obesity and smoking contributed substantially to the inequalities. Great educational inequalities exist in various measures of mobility. Common chronic diseases, obesity, smoking and workload appeared to be the main pathways from low education to mobility limitations. General health promotion using methods that also yield good results in the lowest-educational groups is thus a good strategy to reduce the disparities in mobility.


Within- and between-subject variation in commonly measured anthropometric and biochemical variables.
The biological variation of some commonly assessed metabolic variables in healthy subjects has not been studied extensively. The aim of the study was to assess, in 12 healthy subjects (6 male and 6 female; mean (SD) age; 22.7 (1.5) years) following an overnight fast, the day-to-day variation of body fat (impedance method), triglycerides, nonesterified fatty acid (NEFAs), glycerol, 3-hydroxybutyrate (3-OHB), lactate, glucose, insulin (RIA), C-peptide, and glucagon on 12 consecutive days. Between- and within-subject coefficients of variation (CVG and CVW) were estimated using a random effects analysis of variance, and assay variation was subtracted to give the coefficient of within-subject biological variation (CVI). Individuality indices were calculated as CVW/CVG. The overall means, CVI, and individuality indices were as follows: for body fat, 24.2%, 10%, and 0.3; for triglycerides, 0.61 mmol/L, 21%, and 1.1; for NEFAs, 376 micromol/L, 45%, and 1.4; for glycerol, 48 micromol/L, 36%, and 0.8; for 3-OHB, 43 micromol/L, 61%, and 1.5; for lactate, 0.88 mmol/L, 31%, and 1.1; for glucose, 4.9 mmol/L, 4.8%, and 0.7; for insulin, 52 pmol/L, 26%, and 1.0; for C-peptide, 0.39 nmol/L, 24%, and 0.9; and for glucagon, 53 ng/L, 19%, and 0.8. The data presented here are necessary for the evaluation of several important metabolic variables in individual and group studies. The biological variation of some metabolites makes it difficult to characterize the status of healthy subjects with a single measurement.


Diagnostic and Prognostic Biomarkers of Adrenal Cortical Carcinoma.
The diagnosis of low-grade adrenal cortical carcinoma (ACC) confined to the adrenal gland can be challenging. Although there are diagnostic and prognostic molecular tests for ACC, they remain largely unutilized. We examined the diagnostic and prognostic value of altered reticulin framework and the immunoprofile of biomarkers including IGF-2, proteins involved in cell proliferation and mitotic spindle regulation (Ki67, p53, BUB1B, HURP, NEK2), DNA damage repair (PBK, γ-H2AX), telomere regulation (DAX, ATRX), wnt-signaling pathway (beta-catenin) and PI3K signaling pathway (PTEN, phospho-mTOR) in a tissue microarray of 50 adenomas and 43 carcinomas that were characterized for angioinvasion as defined by strict criteria, Weiss score, and mitotic rate-based tumor grade. IGF-2 and proteins involved in cell proliferation and mitotic spindle regulation (Ki67, p53, BUB1B, HURP, NEK2), DNA damage proteins (PBK, γ-H2AX), regulators of telomeres (DAXX, ATRX), and beta-catenin revealed characteristic expression profiles enabling the distinction of carcinomas from adenomas. Not all biomarkers were informative in all carcinomas. IGF-2 was the most useful biomarker of malignancy irrespective of tumor grade and cytomorphologic features, as juxtanuclear Golgi-pattern IGF-2 reactivity optimized for high specificity was identified in up to 80% of carcinomas and in no adenomas. Loss rather than qualitative alterations of the reticulin framework yielded statistical difference between carcinoma and adenoma. Angioinvasion defined as tumor cells invading through a vessel wall and intravascular tumor cells admixed with thrombus proved to be the best prognostic parameter, predicting adverse outcome in the entire cohort as well as within low-grade ACCs. Low mitotic tumor grade, Weiss score, global loss of DAXX expression, and high phospho-mTOR expression correlated with disease-free survival, but Weiss score and biomarkers failed to predict adverse outcome in low-grade disease. Our results underscore the importance of careful morphologic assessment coupled with ancillary diagnostic and prognostic biomarkers of ACC.


European Competence Network on Mastocytosis (ECNM): 10-year jubilee, update, and future perspectives.
The European Competence Network on Mastocytosis (ECNM) was initiated in 2002 as a multidisciplinary and multinational cooperative approach to increase awareness and to improve diagnosis and therapy of mastocytosis. The network is composed of local centers, physicians, and scientists who have dedicated their work to patients with mastocytosis. A strategic goal of the ECNM is to provide the best available information about the disease to patients and physicians. During the past 10 years, the ECNM has expanded to various countries and contributed successfully to the development of markers, definitions, and standards in the field of mastocytosis. Members of the ECNM organized Annual Meetings in Europe and two Working Conferences on Mastocytosis in Vienna (in 2005 and 2010), and initiated and supported several preclinical and clinical trials. In all these activities, representatives of the ECNM cooperate closely with their US colleagues, with patient-organizations in Europe and in the USA, and with other scientific networks. The ECNM also launched a mastocytosis registry that has been activated in 2012. Using the central database of this registry, cooperative multicenter studies, which should include sufficient numbers of patients and robust evaluations, will be conducted. These studies will increase our knowledge about optimal management and therapy of patients with mastocytosis in the future.