Relations among home- and community-based services investment and nursing home rates of use for working-age and older adults: a state-level analysis.
I examined state-level rates of nursing home use for the period from 2000 to 2007. I used multivariate fixed-effects models to examine associations between state sociodemographic, economic, supply, and programmatic characteristics and rates of use. Nursing home use declined among older adults (aged ≥65 years) in more than two thirds of states and the District of Columbia but increased among older working-age adults (aged 31-64 years) in all but 2 states. State characteristics associated with these trends differed by age group. Although relatively greater state investment in Medicaid home- and community-based services coupled with reduced nursing home capacity was associated with reduced rates of nursing home care for adults aged 65 years and older, neither characteristic was associated with use among older working-age adults. Their use was associated with state sociodemographic characteristics, as well as chronic disease prevalence. Policy efforts to expand home- and community-based services and to reduce nursing facility capacity appear warranted. To more fully extend the Supreme Court's Olmstead decision's promise to older working-age adults, additional efforts to understand factors driving their increasing use are required.


Self-Healing Nanofiber-Reinforced Polymer Composites. 1. Tensile Testing and Recovery of Mechanical Properties.
The present work aims at development of self-healing materials capable of partially restoring their mechanical properties under the conditions of prolonged periodic loading and unloading, which is characteristic, for example, of aerospace applications. Composite materials used in these and many other applications frequently reveal multiple defects stemming from their original inhomogeneity, which facilitates microcracking and delamination at ply interfaces. Self-healing nanofiber mats may effectively prevent such damage without compromising material integrity. Two types of core-shell nanofibers were simultaneously electrospun onto the same substrate in order to form a mutually entangled mat. The first type of core-shell fibers consisted of resin monomer (dimethylsiloxane) within the core and polyacrylonitrile within the shell. The second type of core-shell nanofibers consisted of cure (dimethyl-methyl hydrogen-siloxane) within the core and polyacrylonitrile within the shell. These mutually entangled nanofiber mats were used for tensile testing, and they were also encased in polydimethylsiloxane to form composites that were also subsequently subjected to tensile testing. During tensile tests, the nanofibers can be damaged in stretching up to the plastic regime of deformation. Then, the resin monomer and cure was released from the cores and the polydimethylsiloxane resin was polymerized, which might be expected to result in the self-healing properties of these materials. To reveal and evaluate the self-healing properties of the polyacrylonitrile-resin-cure nanofiber mats and their composites, the results were compared to the tensile test results of the monolithic polyacrylonitrile nanofiber mats or composites formed by encasing polyacrylonitrile nanofibers in a polydimethylsiloxane matrix. The latter do not possess self-healing properties, and indeed, do not recover their mechanical characteristics, in contrast to the polyacrylonitrile-resin-cure nanofiber mats and the composites reinforced by such mats. This is the first work, to the best of our knowledge, where self-healing nanofibers and composites based on them were developed, tested, and revealed restoration of mechanical properties (stiffness) in a 24 h rest period at room temperature.


Nerve fibroblast impact on Schwann cell behavior.
In order to reveal non-neuronal cell interactions after peripheral nerve lesions, we began to analyze the impact of sciatic nerve fibroblasts on Schwann cells in vitro. Both cell types are considered to have opposite effects on axonal regeneration. Few data are available on how repulsive nerve fibroblasts affect neuritotrophic Schwann cells and thus might indirectly influence axonal regrowth. Using different culture systems in conjunction with time-lapse video recording, metabolic labeling, pharmacological intervention, RNAi knockdown, Western blotting and RT-PCR analysis, we found that nerve fibroblasts differentially modify the various responses of Schwann cells. In the presence of collagen type IV and heparan sulfate proteoglycan but not of laminin, diffusible fibroblast factors slow down Schwann cell proliferation. In contrast, fibroblast factors increase the migratory activity of Schwann cells without being chemoattractive. One pro-migratory fibroblast factor turned out to be neuregulin. The pro-migratory activity of nerve fibroblasts and of recombinant neuregulin-1beta1 can be counteracted by neuregulin-specific pharmacological intervention and by neuregulin RNA interference. We show for the first time that nerve fibroblasts play antagonistic and agonistic roles for Schwann cells in a context-dependent manner. The data shed light on cellular mechanisms and have implications for some neuro-tissue engineering strategies.


Manipulating the loss in electromagnetic cloaks for perfect wave absorption.
We examine several ways to manipulate the loss in electro-magnetic cloaks, based on transformation electromagnetics. It is found that, by utilizing inherent electric and magnetic losses of metamaterials, perfect wave absorption can be achieved based on several popular designs of electromagnetic cloaks. A practical implementation of the absorber, consisting of ten discrete layers of metamaterials, is proposed. The new devices demonstrate super-absorptivity over a moderate wideband range, suitable for both microwave and optical applications. It is corroborated that the device is functional with a subwavelength thickness and, hence, advantageous compared to the conventional absorbers.


Oat beta-glucan effects on neutrophil respiratory burst activity following exercise.
Fatiguing exercise has been associated with a decrease in certain functions of neutrophils, whereas moderate exercise has generally been associated with an increase. Consumption of oat beta-glucan (ObetaG), a soluble fiber and mild immune system enhancer, may offset the immunosuppression associated with intense training and perhaps further enhance the benefits of moderate exercise. To test the effects of ObetaG consumption on neutrophil function and number after both moderate and fatiguing exercise. Male mice were assigned to one of six treatment groups. Fatiguing exercise mice (Ftg-ObetaG and Ftg-H2O) ran to volitional fatigue on a treadmill for three consecutive days, and moderate exercise mice (Mod-ObetaG and Mod-H2O) ran for six consecutive days for 1 h. Control mice (Con-ObetaG and Con-H2O) were exposed to the treadmill environment but did not run. ObetaG was consumed in the drinking water (approximately 0.6 mL x d(-1)) for 10 consecutive days. After rest or exercise on the last day of training, mice were given a 1-mL i.p. injection of thioglycollate. Mice were sacrificed 3 h later; neutrophils were harvested from the peritoneal cavity and counted, and their respiratory burst activity was measured using flow cytometry. Both moderate exercise and ObetaG increased neutrophil burst activity, whereas fatiguing exercise had no effect. Neutrophil number was increased by fatiguing exercise and ObetaG, but not moderate exercise. There were no additive effects of exercise and ObetaG on either of these variables. These data suggest that although not additive in their effects, both ObetaG and exercise can alter overall neutrophil respiratory burst activity (number and/or function), but only ObetaG increased both number and function, which may have important ramifications for defense against infection.


Occipital nerve block for the short-term preventive treatment of migraine: A randomized, double-blinded, placebo-controlled study.
Occipital nerve (ON) injections with corticosteroids and/or local anesthetics have been employed for the acute and preventive treatment of migraine for decades. However, to date there is no randomized, placebo-controlled evidence to support the use of occipital nerve block (ONB) for the prevention of migraine. The objective of this article is to determine the efficacy of ONB with local anesthetic and corticosteroid for the preventive treatment of migraine. Patients between 18 and 75 years old with ICHD-II-defined episodic (> 1 attack per week) or chronic migraine (modified ICHD-II as patients with > 10 days with consumption of acute medications were permitted into the study) were randomized to receive either 2.5 ml 0.5% bupivacaine plus 0.5 ml (20 mg) methylprednisolone over the ipsilateral (unilateral headache) or bilateral (bilateral headache) ON or 2.75 ml normal saline plus 0.25 ml 1% lidocaine without epinephrine (placebo). Patients completed a one-month headache diary prior to and after the double-blind injection. The primary outcome measure was defined as a 50% or greater reduction in the frequency of days with moderate or severe migraine headache in the four-week post-injection compared to the four-week pre-injection baseline period. Thirty-four patients received active and 35 patients received placebo treatment. Because of missing data, the full analysis of 33 patients in the active and 30 patients in the placebo group was analyzed for efficacy. In the active and placebo groups respectively, the mean frequency of at least moderate (mean 9.8 versus 9.5) and severe (3.6 versus 4.3) migraine days and acute medication days (7.9 versus 10.0) were not substantially different at baseline. The percentage of patients with at least a 50% reduction in the frequency of moderate or severe headache days was 30% for both groups (10/30 vs nine of 30, Δ 0.00, 95% CI -0.22 to 0.23). Greater ONB does not reduce the frequency of moderate to severe migraine days in patients with episodic or chronic migraine compared to placebo.The study was registered with ClinicalTrial.gov (NCT00915473).


Mapping differentiation pathways from hematopoietic stem cells using Flk2/Flt3 lineage tracing.
Genetic fate-mapping approaches provide a unique opportunity to assess differentiation pathways under physiological conditions. We have recently employed a lineage tracing approach to define hematopoietic differentiation pathways in relation to expression of the tyrosine kinase receptor Flk2.1 Based on our examination of reporter activity across all stem, progenitor and mature populations in our Flk2-Cre lineage model, we concluded that all mature blood lineages are derived through a Flk2+ intermediate, both at steady-state and under stress conditions. Here, we re-examine in depth our initial conclusions and perform additional experiments to test alternative options of lineage specification. Our data unequivocally support the conclusion that onset of Flk2 expression results in loss of self-renewal but preservation of multilineage differentiation potential. We discuss the implications of these data for defining stem cell identity and lineage potential among hematopoietic populations.


Alcohol consumption and bone mineral density in elderly women.
Findings regarding alcohol consumption and bone mineral density (BMD) in elderly women have been inconsistent. The objective of the present study was to explore the association of alcohol intake with BMD in elderly women. This cohort study included women from the population-based Kuopio Osteoporosis Risk Factor and Prevention - Fracture Prevention Study (OSTPRE-FPS). Alcohol intake and potential confounders were assessed at baseline and after 3 years of follow-up using a lifestyle questionnaire. In addition, an FFQ was distributed in the third year to measure dietary intake, including alcohol. Women underwent BMD measurements at the femoral neck and lumbar spine at baseline and after 3 years of follow-up. Kuopio Province, Finland. Three hundred elderly women (mean age 67·8 years) who provided both BMD measurements and FFQ data. Alcohol consumption estimated from the FFQ and lifestyle questionnaire was significantly associated with BMD at both measurement sites after adjustment for potential confounders, including lifestyle and dietary factors (P < 0·05). Using the FFQ, women drinking >3 alcoholic drinks/week had significantly higher BMD than abstainers, 12·0 % at the femoral neck and 9·2 % at the lumbar spine. Results based on the lifestyle questionnaire showed higher BMD values for all alcohol-consuming women at the femoral neck and for women drinking 1-3 alcoholic beverages/week at the lumbar spine, compared with non-users. The results from OSTPRE-FPS suggest that low to moderate alcohol intake may exert protective effects on bone health in elderly women.


Influenza vaccination coverage rates in five European countries during season 2006/07 and trends over six consecutive seasons.
The objectives of the survey were to identify the level of influenza vaccination coverage in five European countries between 2001 and 2007, to understand the drivers and barriers to vaccination, to assess vaccination intentions for the winter 2007/08 as well as major encouraging factors for vaccination. Between 2001 and 2007, representative household surveys were performed with telephone or mailed (France) interviews of individuals aged 14 and above. The questionnaire used in the UK, Germany, Italy, France and Spain was essentially the same in all seasons. The data were subsequently pooled. Four target groups were defined for the analysis: 1) persons aged 65 years and over; 2) persons working in the medical field; 3) chronically ill persons and 4) combined target group composed of individuals belonging to one or more of the previous groups 1, 2 or 3. In 2006/07, vaccination coverage was, 25.0% in UK, 27.4% in Germany, 21.8% in Spain, 24.2% in France and 24.4% in Italy. During six influenza seasons (2001-2007), vaccination coverage showed a slight positive trend in the five countries (p < or = 0.0001). In the elderly (> or = 65 years), across all countries, no significant trend was seen; the vaccination rate decreased non-significantly from a peak of 64.2% in season 2005/06 to 61.1% in season 2006/07. The most frequent reason for getting vaccinated was a recommendation by the family doctor or nurse (51%), and this was also perceived as the major encouraging factor for vaccination (61%). The main reason for not getting vaccinated was feeling unlikely to catch the flu (36%). In the UK, Germany and Spain, influenza vaccination coverage rates in season 2006/07 dropped slightly compared to the previous season. However, a trend of increasing vaccination coverage was observed from 2001/02 to 2006/07 across Europe. The family doctor is the major source of encouragement for individuals getting vaccinated. Efforts to overcome the barriers to vaccination need to be put in place to reach the WHO objective of 75% coverage in the elderly by 2010. This is a major challenge to be faced by governments, healthcare workers and healthcare organisations.


Transcriptional control by adenovirus E1A conserved region 3 via p300/CBP.
The human adenovirus type 5 (HAdV-5) E1A 13S oncoprotein is a potent regulator of gene expression and is used extensively as a model for transcriptional activation. It possesses two independent transcriptional activation domains located in the N-terminus/conserved region (CR) 1 and CR3. The protein acetyltransferase p300 was previously identified by its association with the N-terminus/CR1 portion of E1A and this association is required for oncogenic transformation by E1A. We report here that transcriptional activation by 13S E1A is inhibited by co-expression of sub-stoichiometric amounts of the smaller 12S E1A isoform, which lacks CR3. Transcriptional inhibition by E1A 12S maps to the N-terminus and correlates with the ability to bind p300/CBP, suggesting that E1A 12S is sequestering this limiting factor from 13S E1A. This is supported by the observation that the repressive effect of E1A 12S is reversed by expression of exogenous p300 or CBP, but not by a CBP mutant lacking actyltransferase activity. Furthermore, we show that transcriptional activation by 13S E1A is greatly reduced by siRNA knockdown of p300 and that CR3 binds p300 independently of the well-characterized N-terminal/CR1-binding site. Importantly, CR3 is also required to recruit p300 to the adenovirus E4 promoter during infection. These results identify a new functionally significant interaction between E1A CR3 and the p300/CBP acetyltransferases, expanding our understanding of the mechanism by which this potent transcriptional activator functions.


Coupling of 10-methacryloyloxydecyldihydrogenphosphate to tetragonal zirconia: Effect of pH reaction conditions on coordinate bonding.
Identification of the mechanism of chemical coupling of a phosphate ester monomer to zirconia via a computational modeling approach enables materials scientists to design new coupling agents that can resist hydrolytic degradation of bonds made by methacrylate resins to zirconia. We investigated the possibility of chemical bonding between 10-methacryloyloxydecyldihydrogenphosphate (MDP) and tetragonal zirconia, and the effect of pH reaction conditions on such prospective chemical bonds. A tetragonal zirconia crystal model was created. An "Our-own N-layered integrated molecular orbital and molecular mechanics" (ONIOM) method was used to simulate two potential configurations of the MDP-ZrO2 system: double-coordinate and single-coordinate. Thermodynamic calculations were used to ascertain if the reaction could proceed spontaneously and to compare the stability of the two possible configurations. Short-term testing of shear bond strength (SBS) was done to evaluate bonding improvement of MDP to alumina-sandblasted zirconia surfaces in neutral, acidic or alkaline environments. Digital models of coordinate bonds between MDP and tetragonal zirconia were constructed. Thermodynamic calculations indicated that the Gibbs free energy for forming double-coordinate and single-coordinate configurations were -461.2kJ/mol and -450.9kJ/mol, respectively. Equilibrium constants for the double-coordinate and single-coordinate configurations were 6.4×10(80) and 9.8×10(78), respectively. Application of MDP in alkaline conditions showed the highest SBS, whereas acid conditions resulted in lower SBS. MDP can establish a "true" chemical bond with zirconia spontaneously. The double-coordinate configuration was identified to be more energetically favorable than the single-coordinate configuration for the coupling of MDP to zirconia. Alkaline conditions may positively affect formation of MDP-ZrO2 coordination bonds.


Left atrial strain: a new parameter for assessment of left ventricular filling pressure.
In order to obtain accurate diagnosis, treatment and prognostication in many cardiac conditions, there is a need for assessment of left ventricular (LV) filling pressure. While systole depends on ejection function of LV, diastole and its disturbances influence filling function and pressures. The commonest condition that represents the latter is heart failure with preserved ejection fraction in which LV ejection is maintained, but diastole is disturbed and hence filling pressures are raised. Significant diastolic dysfunction results in raised LV end-diastolic pressure, mean left atrial (LA) pressure and pulmonary capillary wedge pressure, all referred to as LV filling pressures. Left and right heart catheterization has traditionally been used as the gold standard investigation for assessing these pressures. More recently, Doppler echocardiography has taken over such application because of its noninvasive nature and for being patient friendly. A number of indices are used to achieve accurate assessment of filling pressures including: LV pulsed-wave filling velocities (E/A ratio, E wave deceleration time), pulmonary venous flow (S wave and D wave), tissue Doppler imaging (E' wave and E/E' ratio) and LA volume index. LA longitudinal strain derived from speckle tracking echocardiography (STE) is also sensitive in estimating intracavitary pressures. It is angle-independent, thus overcomes Doppler limitations and provides highly reproducible measures of LA deformation. This review examines the application of various Doppler echocardiographic techniques in assessing LV filling pressures, in particular the emerging role of STE in assessing LA pressures in various conditions, e.g., HF, arterial hypertension and atrial fibrillation.


Non-invasive assessment of Alzheimer's disease neurofibrillary pathology using 18F-THK5105 PET.
Non-invasive imaging of tau pathology in the living brain would be useful for accurately diagnosing Alzheimer's disease, tracking disease progression, and evaluating the treatment efficacy of disease-specific therapeutics. In this study, we evaluated the clinical usefulness of a novel tau-imaging positron emission tomography tracer 18F-THK5105 in 16 human subjects including eight patients with Alzheimer's disease (three male and five females, 66-82 years) and eight healthy elderly controls (three male and five females, 63-76 years). All participants underwent neuropsychological examination and 3D magnetic resonance imaging, as well as both 18F-THK5105 and 11C-Pittsburgh compound B positron emission tomography scans. Standard uptake value ratios at 90-100 min and 40-70 min post-injection were calculated for 18F-THK5105 and 11C-Pittsburgh compound B, respectively, using the cerebellar cortex as the reference region. As a result, significantly higher 18F-THK5105 retention was observed in the temporal, parietal, posterior cingulate, frontal and mesial temporal cortices of patients with Alzheimer's disease compared with healthy control subjects. In patients with Alzheimer's disease, the inferior temporal cortex, which is an area known to contain high densities of neurofibrillary tangles in the Alzheimer's disease brain, showed prominent 18F-THK5105 retention. Compared with high frequency (100%) of 18F-THK5105 retention in the temporal cortex of patients with Alzheimer's disease, frontal 18F-THK5105 retention was less frequent (37.5%) and was only observed in cases with moderate-to-severe Alzheimer's disease. In contrast, 11C-Pittsburgh compound B retention was highest in the posterior cingulate cortex, followed by the ventrolateral prefrontal, anterior cingulate, and superior temporal cortices, and did not correlate with 18F-THK5105 retention in the neocortex. In healthy control subjects, 18F-THK5105 retention was ∼10% higher in the mesial temporal cortex than in the neocortex. Notably, unlike 11C-Pittsburgh compound B, 18F-THK5105 retention was significantly correlated with cognitive parameters, hippocampal and whole brain grey matter volumes, which was consistent with findings from previous post-mortem studies showing significant correlations of neurofibrillary tangle density with dementia severity or neuronal loss. From these results, 18F-THK5105 positron emission tomography is considered to be useful for the non-invasive assessment of tau pathology in the living brain. This technique would be applicable to the longitudinal evaluation of tau deposition and allow a better understanding of the pathophysiology of Alzheimer's disease.


Antioxidant, ACE-inhibitory and antimicrobial activity of fermented goat milk: activity and physicochemical property relationship of the peptide components.
Increasing evidence on goat milk and the health benefits of its derived products beyond its nutritional value show its potential as a functional food. In this study, goat milk fractions were tested for their total antioxidant capacity using different methods (ORAC, ABTS, DPPH and FRAP), as well as their angiotensin-I-converting-enzyme inhibitory and antimicrobial (against Escherichia coli and Micrococcus luteus) activities. Different whey fractions (whey, cation exchange membrane permeate P and retentate R) of two fermented skimmed goat milks (ultrafiltered goat milk fermented with the classical starter bacteria or with the classical starter plus the Lactobacillus plantarum C4 probiotic strain) were assessed. Additionally, P fractions were divided into two sub-fractions after being passed through a 3 kDa cut-off membrane: (a) the permeate with peptides of MW <3 kDa (P < 3); and (b) the retentate with peptides and proteins of MW >3 kDa (P > 3). No differences in biological activities were observed between the two fermented milks. However, the biological peptides present in the P < 3 fraction showed the highest total antioxidant capacity (for the ORAC assay) and angiotensin-I-converting-enzyme inhibitory activity. Those present in the R fraction showed the highest total antioxidant capacity against ABTS˙+ and DPPH˙ radicals. Some antimicrobial activity against E. coli was observed for the fermented milk containing the probiotic, which could be due to some peptides being released by the probiotic strain. In conclusion, small and non-basic bioactive peptides could be responsible for most of the angiotensin-I-converting-enzyme inhibitory and antioxidant activities. These findings reinforce the potential benefits of the consumption of fermented goat milk in the prevention of cardiovascular diseases associated with oxidative stress and hypertension.


The use of oxybutynin for treating facial hyperhidrosis.
Facial hyperhidrosis is a disease that may lead patients to serious emotional disturbances. Video-assisted thoracic sympathectomy provides excellent resolution of facial hyperhidrosis, but is associated with certain complications. The most frequent and important complication is compensatory hyperhidrosis. Especially in patients who have undergone resection of the second thoracic ganglion, the risk of severe compensatory hyperhidrosis is higher, which may cause dissatisfaction with the procedure. The aim of this study was to evaluate the efficacy of the use of low doses of oxybutynin in treating facial hyperhidrosis as well as the level of patient satisfaction with its use. 25 patients with facial hyperhidrosis were treated with oxybutynin. The patients underwent 2 evaluations: before and after treatment. These evaluations were used to assess the patients' clinical improvement and quality of life. We observed that more than 75% of the patients evolved with an improvement in facial hyperhidrosis, and 52% of them presented a great improvement. Treatment of facial hyperhidrosis with oxybutynin is a good alternative to sympathectomy, since it presents good results and improves quality of life, in addition to not exposing patients to the risk of experiencing the side effects of sympathectomy.


Identification of progressors in osteoarthritis by combining biochemical and MRI-based markers.
At present, no disease-modifying osteoarthritis drugs (DMOADS) are approved by the FDA (US Food and Drug Administration); possibly partly due to inadequate trial design since efficacy demonstration requires disease progression in the placebo group. We investigated whether combinations of biochemical and magnetic resonance imaging (MRI)-based markers provided effective diagnostic and prognostic tools for identifying subjects with high risk of progression. Specifically, we investigated aggregate cartilage longevity markers combining markers of breakdown, quantity, and quality. The study included healthy individuals and subjects with radiographic osteoarthritis. In total, 159 subjects (48% female, age 56.0 +/- 15.9 years, body mass index 26.1 +/- 4.2 kg/m2) were recruited. At baseline and after 21 months, biochemical (urinary collagen type II C-telopeptide fragment, CTX-II) and MRI-based markers were quantified. MRI markers included cartilage volume, thickness, area, roughness, homogeneity, and curvature in the medial tibio-femoral compartment. Joint space width was measured from radiographs and at 21 months to assess progression of joint damage. Cartilage roughness had the highest diagnostic accuracy quantified as the area under the receiver-operator characteristics curve (AUC) of 0.80 (95% confidence interval: 0.69 to 0.91) among the individual markers (higher than all others, P < 0.05) to distinguish subjects with radiographic osteoarthritis from healthy controls. Diagnostically, cartilage longevity scored AUC 0.84 (0.77 to 0.92, higher than roughness: P = 0.03). For prediction of longitudinal radiographic progression based on baseline marker values, the individual prognostic marker with highest AUC was homogeneity at 0.71 (0.56 to 0.81). Prognostically, cartilage longevity scored AUC 0.77 (0.62 to 0.90, borderline higher than homogeneity: P = 0.12). When comparing patients in the highest quartile for the longevity score to lowest quartile, the odds ratio of progression was 20.0 (95% confidence interval: 6.4 to 62.1). Combination of biochemical and MRI-based biomarkers improved diagnosis and prognosis of knee osteoarthritis and may be useful to select high-risk patients for inclusion in DMOAD clinical trials.


Grampian Health Board's joint drug formulary.
To develop a model for creating a joint general practice-hospital formulary, using the example of ulcer healing drugs. A joint formulary development group produced draft guidelines based on an earlier hospital formulary, which were sent to interested local general practitioners for consultation. Revised guidelines were then drawn up and forwarded to the health board's medicines committee for approval and distribution. Grampian Health Board. Nine members of joint formulary development group plus local general practitioners who were invited to comment on a list of 11 ulcer healing drugs. Degree of coincidence of drugs selected by hospital doctors and general practitioners. The ulcer healing drugs selected by the panel of general practitioners and by hospital doctors were highly coincident. The cost of one day's treatment with drugs varied considerably between hospital and general practice--for example, one drug cost 46p in hospital and 1 pounds in general practice and another cost 1.26 pounds in hospital and 1.01 pounds in general practice. Overall, six drugs cost more in hospital and five cost more in general practice. A joint formulary for use in hospitals and general practice in a health board can be devised fairly simply by consultation as virtually the same drugs are used in both types of practice. It should influence the health board's expenditure on drugs and affect the choice of drugs when a patient is discharged from hospital or is referred to any hospital in the region.


How to link theory and experiment for single-chain magnets beyond the Ising model: magnetic properties modeled from ab initio calculations of molecular fragments.
Magnetic properties of coordination polymers like single-chain magnets (SCMs) are based on magnetic domains, which are formed due to magnetic exchange between neighboring anisotropic spin centers. However, the computational restrictions imposed by the high level of theory needed for an adequate ab initio quantum mechanical treatment on the basis of multi-reference methods for these systems limit the feasibility of such calculations to mononuclear fragments as appropriate structural cutouts for the metal centers along the chains. Hence, results from such calculations describe single-ion properties and cannot be directly correlated with experimental data representing magnetic domains. We present a theoretical approach based on n-membered Ising-spin rings with n = 3-12, which allows us to simulate magnetic domains and to derive important magnetic properties for SCM compounds. Magnetic exchange, which is not provided by calculations of mononuclear fragments, is obtained by fitting the theoretical magnetic susceptibility against experimental data. The presented approach is tested for cobalt(ii)-based SCMs with three types of repeating sequences, which differ in nuclearity and symmetry. The magnetic parameters derived using the presented approach were found to be in good agreement with the experimental data. Moreover, the energy spectra obtained for the three test cases using the presented approach are characteristic of a deviation of the individual systems from the ideal Ising behavior. An extrapolation technique towards larger systems (n > 12) is presented which can provide information on the statistical mean length of the magnetic domains in the three investigated SCM compounds.


FFCA: a feasibility-based method for flux coupling analysis of metabolic networks.
Flux coupling analysis (FCA) is a useful method for finding dependencies between fluxes of a metabolic network at steady-state. FCA classifies reactions into subsets (called coupled reaction sets) in which activity of one reaction implies activity of another reaction. Several approaches for FCA have been proposed in the literature. We introduce a new FCA algorithm, FFCA (Feasibility-based Flux Coupling Analysis), which is based on checking the feasibility of a system of linear inequalities. We show on a set of benchmarks that for genome-scale networks FFCA is faster than other existing FCA methods. We present FFCA as a new method for flux coupling analysis and prove it to be faster than existing approaches. A corresponding software tool is freely available for non-commercial use at http://www.bioinformatics.org/ffca/.


How to discriminate contamination from bloodstream infection due to coagulase-negative staphylococci: a prospective study with 654 patients.
Coagulase-negative staphylococci (CoNS) are frequent contaminants of blood cultures. We aimed to evaluate the systemic inflammatory response syndrome (SIRS) criteria in patients with CoNS bacteraemia for discrimination between true bloodstream infection (BSI) and contamination. Prospective evaluation was carried out of clinical and laboratory parameters in adults with at least one positive blood culture with CoNS at the University Hospital of Basel between 2003 and 2007. Of 3060 positive blood cultures, 654 episodes of CoNS bacteraemia were identified. Of these, 232 (35%) were considered to be true BSI and 422 (65%) were considered to be contamination. Overall, 80% of study participants had at least one SIRS criterion, fever being the most common, and 49% had at least two SIRS criteria. In the multivariate analysis, independent predictors of BSI were fever or hypothermia (OR 2.93, 95% CI 1.91-4.5), tachycardia (OR 2.29, 95% CI 1.50-3.50), tachypnoea (OR 2.4, 95% CI 1.30-4.43), leucocytosis or leucopenia (OR 4.15, 95% CI 2.17-6.36) and the presence of a central venous line (OR 5.38, 95% CI 3.25-8.88). The probability of BSI increased with each additional SIRS criterion, ranging from 42.4% in patients with only one SIRS criterion to 56.7% for those with two criteria, and 72.3% for patients with three SIRS criteria. A positive blood culture with CoNS most likely represents true BSI if the patient has at least three SIRS criteria or two SIRS criteria and a central venous catheter. These simple bedside criteria may guide decision to treat, decreasing the use of glycopeptides.


Fibrinogen, viscosity and white blood cell count predict myocardial, but not cerebral infarction: evidence from the Caerphilly and Speedwell cohort.
Fibrinogen, plasma viscocity, and the white blood cell count predict ischaemic heart disease, but there is less certainty for their predictive power for ischaemic stroke. Studying stroke and ischaemic heart disease in the same cohort prospectively allows comparison of predictive strengths. The Caerphilly and Speedwell cohorts consist of a population sample of 4,860 men aged 45-59 years at recruitment who had baseline measurements of fibrinogen, plasma viscosity, and white blood cell counts. After 15-19 years of follow-up, men in the two cohorts experienced 312 ischaemic strokes and 557 ischaemic heart disease events. Mean fibrinogen, plasma viscosity and white blood cell counts differed significantly after adjustment for confounding factors between men with and without ischaemic heart disease, 0.25 g/l (95% CIs 0.1 8-0.32); 0.036 cp (95% CIs 0.027-0.044); 0.67 x 10(9)/l (95% CIs 0.50-0.84) respectively. The same measurements showed no significant differences after adjustment for the same confounding factors for men with and without ischaemic stroke, 0.05 g/l (95% CIs -0.04-0.14); 0.008 cp (95% CIs -0.003-0.019); 0.16 x 10(9)/l (95% CIs -0.06-0.38) respectively.


Severe folate-deficiency pancytopenia.
Folate-deficiency anaemia occurs in about 4 per 100 000 people, although severe cases causing moderate pancytopenia are rarer. We present the case of a significant folate deficiency in a 50-year-old alcoholic with a background of mild liver impairment and recurrent nasal and rectal bleeding. Her blood tests showed profound macrocytic anaemia with haemoglobin 2.6 g/dl, leucopoenia with white cell count 3.2 × 10(9)/litre and thrombocytopenia with platelets 17 × 10(9)/litre. Serum folate was 0.8 ng/ml (normal 2.5-13.5 ng/ml) confirming severe deficiency. Despite these life-threatening results, the patient was stable, alert and was keen to avoid admission. Medical management of the anaemia included slow transfusion of red cells and one unit of platelets in view of haemorrhagic symptoms, two injections of vitamin B12 while awaiting assays and oral folic acid. A rapid improvement in the leucopoenia and thrombocytopenia resulted and no additional complications were encountered.


On the temporal dynamics of sign production: An ERP study in Catalan Sign Language (LSC).
This study investigates the temporal dynamics of sign production and how particular aspects of the signed modality influence the early stages of lexical access. To that end, we explored the electrophysiological correlates associated to sign frequency and iconicity in a picture signing task in a group of bimodal bilinguals. Moreover, a subset of the same participants was tested in the same task but naming the pictures instead. Our results revealed that both frequency and iconicity influenced lexical access in sign production. At the ERP level, iconicity effects originated very early in the course of signing (while absent in the spoken modality), suggesting a stronger activation of the semantic properties for iconic signs. Moreover, frequency effects were modulated by iconicity, suggesting that lexical access in signed language is determined by the iconic properties of the signs. These results support the idea that lexical access is sensitive to the same phenomena in word and sign production, but its time-course is modulated by particular aspects of the modality in which a lexical item will be finally articulated.


Endothelial transdifferentiated phenotype and cell-cycle kinetics of AIDS-associated Kaposi sarcoma cells.
The nature of Kaposi sarcoma (KS) (vascular malignancy vs. discordant angiogenesis) and lineage of the progenitor cell remain unclear. Therefore, AIDS-KS enzyme isolate cultures were prepared from excised skin lesions. Endothelial marker positivity for Factor VIII related antigen (F8RAg), Ulex europaeus agglutinin (UEA), and angiotensin-converting enzyme (ACE) were determined by fluorescence microscopy (FM) and flow cytometry (FCM). DNA cell-cycle analysis was performed using FCM. KS lesions showed large thick-walled channels (F8RAg and UEA strongly +), narrow vascular slits and thin-walled lakes (F8RAg and UEA weakly +), and non-prominent spindle cells (F8RAg and UEA almost uniformly negative). KS cultures yielded heterogenous populations of spindle, stellate, and flattened endothelial-like cells, displaying positivity for F8RAg (64 +/- 3%; mean +/- SE), UEA (40 +/- 9%), and ACE (81 +/- 9%). When injected subcutaneously in the nude mouse these cells failed to produce tumors. During contact inhibition induced quiescence, KS cultures exhibited a high G2M (18 +/- 3%) compared to non-KS (7 +/- 4%; p < 0.04), evidence of an altered proliferative potential consistent with a transdifferentiated or transformed phenotype.


Risk of cardiovascular disease morbidity and mortality in frail and pre-frail older adults: Results from a meta-analysis and exploratory meta-regression analysis.
Frailty is common and associated with poorer outcomes in the elderly, but its role as potential cardiovascular disease (CVD) risk factor requires clarification. We thus aimed to meta-analytically evaluate the evidence of frailty and pre-frailty as risk factors for CVD. Two reviewers selected all studies comparing data about CVD prevalence or incidence rates between frail/pre-frail vs. robust. The association between frailty status and CVD in cross-sectional studies was explored by calculating and pooling crude and adjusted odds ratios (ORs) ±95% confidence intervals (CIs); the data from longitudinal studies were pooled using the adjusted hazard ratios (HRs). Eighteen cohorts with a total of 31,343 participants were meta-analyzed. Using estimates from 10 cross-sectional cohorts, both frailty and pre-frailty were associated with higher odds of CVD than robust participants. Longitudinal data were obtained from 6 prospective cohort studies. After a median follow-up of 4.4 years, we identified an increased risk for faster onset of any-type CVD in the frail (HR=1.70 [95%CI, 1.18-2.45]; I2=66%) and pre-frail (HR=1.23 [95%CI, 1.07-1.36]; I2=67%) vs. robust groups. Similar results were apparent for time to CVD mortality in the frail and pre-frail groups. In conclusion, frailty and pre-frailty constitute addressable and independent risk factors for CVD in older adults.


Identification of two regions in apolipoprotein B100 that are exposed on the cytosolic side of the endoplasmic reticulum membrane.
Protease protection assays of apolipoprotein B100 (apoB) in digitonin-permeabilized HepG2 cells indicated that multiple domains of apoB are exposed to the cytosol through an extensive portion of the secretory pathway. The intracellular orientation of apoB in the secretory pathway was confirmed by immunocytochemistry using antibodies recognizing specific domains of apoB in streptolysin-O (STP-O)- and saponin-permeabilized HepG2 cells. Lumenal epitopes on marker proteins in secretory pathway compartments (p63, p53, and galactosyltransferase) were not stained by antibodies in STP-O-treated cells, but were brightly stained in saponin-treated cells, confirming that internal membranes were not perforated in STP-O-treated cells. An anti-apoB peptide antibody (B4) recognizing amino acids 3221-3240 caused intense staining in close proximity to the nuclear membrane, and less intensely throughout the secretory pathway in STP-O-permeabilized cells. Staining with this antibody was similar in STP-O- and saponin-treated cells, indicating that this epitope in apoB is exposed to the cytosol at the site of apoB synthesis and throughout most of the remaining secretory pathway. Similar results indicating a cytosolic orientation were obtained with monoclonal antibody CC3.4, which recognizes amino acids 690-797 (79-91 kD) in apoB. Two polyclonal antibodies made to human LDL and two monoclonal antibodies recognizing amino acids 1878-2148 (D7.2) and 3214-3506 (B1B6) in apoB did not produce a strong reticular signal for apoB in STP-O-treated cells. The anti-LDL and B1B6 antibodies produced almost identical punctate patterns in STP-O-treated cells that overlapped with LAMP-1, a membrane marker for lysosomes. These observations suggest that the B1B6 epitope of apoB is exposed on the surface of the lysosome. The results identify two specific regions in apoB that are exposed to the cytosol in the secretory pathway.


Transperitoneal laparoscopic left versus right live donor nephrectomy: Comparison of outcomes.
Although laparoscopic donor nephrectomy (LDN) is being performed at many centers, there are reservations on the routine use of laparoscopy for harvesting the right kidney due to a perception of technical complexity and increased incidence of allograft failure, renal vein thrombosis and the need for more back-table reconstruction along with increased operative time. We performed a prospective non-randomized comparison of transperitoneal laparoscopic left donor nephrectomy (LLDN) with laparoscopic right donor nephrectomy (RLDN) from August 2008 to May 2013. The operative time, warm ischemia time, intraoperative events, blood loss and post-operative parameters were recorded. The renal recipient parameters, including post-operative creatinine, episodes of acute tubular necrosis (ATN) and delayed graft function were also recorded. A total of 188 LDN were performed between August 2008 and May 2013, including 164 LLDN and 24 RLDN. The demographic characteristics between the two groups were comparable. The operative duration was in favor of the right donor group, while warm ischemia time, estimated blood loss and mean length of hospital stay were similar between the two groups. Overall renal functional outcomes were comparable between the two donor groups, while the recipient outcomes including creatinine at discharge were also comparable. RLDN has a safety profile comparable with LLDN, even in those with complex vascular anatomy, and can be successfully performed by the transperitoneal route with no added morbidity. RLDN requires lesser operative time with comparable morbidity.


A novel method for assessing risks to pollinators from plant protection products using honeybees as a model species.
Pollination is one of the most important ecosystem services in agroecosystems and supports food production. Pollinators are potentially at risk being exposed to pesticides and the main route of exposure is direct contact, in some cases ingestion, of contaminated materials such as pollen, nectar, flowers and foliage. To date there are no suitable methods for predicting pesticide exposure for pollinators, therefore official procedures to assess pesticide risk are based on a Hazard Quotient. Here we develop a procedure to assess exposure and risk for pollinators based on the foraging behaviour of honeybees (Apis mellifera) and using this species as indicator representative of pollinating insects. The method was applied in 13 European field sites with different climatic, landscape and land use characteristics. The level of risk during the crop growing season was evaluated as a function of the active ingredients used and application regime. Risk levels were primarily determined by the agronomic practices employed (i.e. crop type, pest control method, pesticide use), and there was a clear temporal partitioning of risks through time. Generally the risk was higher in sites cultivated with permanent crops, such as vineyard and olive, than in annual crops, such as cereals and oil seed rape. The greatest level of risk is generally found at the beginning of the growing season for annual crops and later in June-July for permanent crops.


Protective effect of low-concentration chlorine dioxide gas against influenza A virus infection.
Influenza virus infection is one of the major causes of human morbidity and mortality. Between humans, this virus spreads mostly via aerosols excreted from the respiratory system. Current means of prevention of influenza virus infection are not entirely satisfactory because of their limited efficacy. Safe and effective preventive measures against pandemic influenza are greatly needed. We demonstrate that infection of mice induced by aerosols of influenza A virus was prevented by chlorine dioxide (ClO(2)) gas at an extremely low concentration (below the long-term permissible exposure level to humans, namely 0.1 p.p.m.). Mice in semi-closed cages were exposed to aerosols of influenza A virus (1 LD(50)) and ClO(2) gas (0.03 p.p.m.) simultaneously for 15 min. Three days after exposure, pulmonary virus titre (TCID(50)) was 10(2.6+/-1.5) in five mice treated with ClO(2), whilst it was 10(6.7+/-0.2) in five mice that had not been treated (P=0.003). Cumulative mortality after 16 days was 0/10 mice treated with ClO(2) and 7/10 mice that had not been treated (P=0.002). In in vitro experiments, ClO(2) denatured viral envelope proteins (haemagglutinin and neuraminidase) that are indispensable for infectivity of the virus, and abolished infectivity. Taken together, we conclude that ClO(2) gas is effective at preventing aerosol-induced influenza virus infection in mice by denaturing viral envelope proteins at a concentration well below the permissible exposure level to humans. ClO(2) gas could therefore be useful as a preventive means against influenza in places of human activity without necessitating evacuation.


Integration between in vivo dosimetry and image guided radiotherapy for lung tumors.
The article reports a feasibility study about the potentiality of an in vivo dosimetry method for the adaptive radiotherapy of the lung tumors treated by 3D conformal radiotherapy techniques (3D CRTs). At the moment image guided radiotherapy (IGRT) has been used for this aim, but it requires taking many periodic radiological images during the treatment that increase workload and patient dose. In vivo dosimetry reported here can reduce the above efforts, alerting the medical staff for the commissioning of new radiological images for an eventual adaptive plan. The in vivo dosimetry method applied on 20 patients makes use of the transit signal St on the beam central axis measured by a small ion chamber positioned on an electronic portal imaging device (EPID) or by the EPID itself. The reconstructed in vivo dosimetry at the isocenter point Diso requires a convolution between the transit signal St and a dose reconstruction factor C that essentially depends on (i) tissue inhomogeneities along the beam central axis and (ii) the in-patient isocenter depth. The C factors, one for every gantry angle, are obtained by processing the patient's computed tomography scan. The method has been recently applied in some Italian centers to check the radiotherapy of pelvis, breast, head, and thorax treatments. In this work the dose reconstruction was carried out in five centers to check the Diso in the lung tumor during the 3D CRT, and the results have been used to detect the interfraction tumor anatomy variations that can require new CT imaging and an adaptive plan. In particular, in three centers a small ion chamber was positioned below the patient and used for the St measurement. In two centers, the St signal was obtained directly by 25 central pixels of an a-Si EPID, equipped with commercial software that enabled its use as a stable detector. A tolerance action level of +/- 6% for every checked beam was assumed. This means that when a difference greater than 6% between the predicted dose by the treatment planning system, Diso,TPS, and the Diso was observed, the clinical action started to detect possible errors. 60% of the patients examined presented morphological changes during the treatment that were checked by the in vivo dosimetry and successively confirmed by the new CT scans. In this work, a patient that showed for all beams Diso values outside the tolerance level, new CT scans were commissioned for an adaptive plan. The lung dose volume histograms (DVHs) for a Diso,TPs=2 Gy for fraction suggested the adaptive plan to reduce the dose in lung tissue. The results of this research show that the dose guided radiotherapy (DGRT) by the Diso reconstruction was feasible for daily or periodic investigation on morphological lung tumor changes. In other words, since during 3D CRT treatments the anatomical lung tumor changes occur frequently, the DGRT can be well integrated with the IGRT.


Single nucleotide polymorphisms associated with susceptibility for development of colorectal cancer: Case-control study in a Basque population.
Given the significant population diversity in genetic variation, we aimed to investigate whether single nucleotide polymorphisms (SNPs) previously identified in studies of colorectal cancer (CRC) susceptibility were also relevant to the population of the Basque Country (North of Spain). We genotyped 230 CRC cases and 230 healthy controls for 48 previously reported CRC-susceptibility SNPs. Only the rs6687758 in DUPS10 exhibited a statistically significant association with CRC risk based on the crude analysis. The rs6687758 AG genotype conferred about 2.13-fold increased risk for CRC compared to the AA genotype. Moreover, we found significant associations in cases between smoking status, physical activity, and the rs6687758 SNP. The results of a Genetic Risk Score (GRS) showed that the risk alleles were more frequent in cases than controls and the score was associated with CRC in crude analysis. In conclusion, we have confirmed a CRC susceptibility locus and the existence of associations between modifiable factors and the rs6687758 SNP; moreover, the GRS was associated with CRC. However, further experimental validations are needed to establish the role of this SNP, the function of the gene identified, as well as the contribution of the interaction between environmental factors and this locusto the risk of CRC.


Impacts of inbreeding on bumblebee colony fitness under field conditions.
Inbreeding and the loss of genetic diversity are known to be significant threats to small, isolated populations. Hymenoptera represent a special case regarding the impact of inbreeding. Haplodiploidy may permit purging of deleterious recessive alleles in haploid males, meaning inbreeding depression is reduced relative to diploid species. In contrast, the impact of inbreeding may be exacerbated in Hymenopteran species that have a single-locus complementary sex determination system, due to the production of sterile or inviable diploid males. We investigated the costs of brother-sister mating in the bumblebee Bombus terrestris. We compared inbred colonies that produced diploid males and inbred colonies that did not produce diploid males with outbred colonies. Mating, hibernation and colony founding took place in the laboratory. Once colonies had produced 15 offspring they were placed in the field and left to forage under natural conditions. The diploid male colonies had a significantly reduced fitness compared to regular inbred and outbred colonies; they had slower growth rates in the laboratory, survived for a shorter time period under field conditions and produced significantly fewer offspring overall. No differences in success were found between non-diploid male inbred colonies and outbred colonies. Our data illustrate that inbreeding exacts a considerable cost in Bombus terrestris through the production of diploid males. We suggest that diploid males may act as indicators of the genetic health of populations, and that their detection could be used as an informative tool in hymenopteran conservation. We conclude that whilst haplodiploids may suffer less inbreeding depression than diploid species, they are still highly vulnerable to population fragmentation and reduced genetic diversity due to the extreme costs imposed by the production of diploid males.


Estimates of emotional and psychometric intelligence: evidence for gender-based stereotypes.
The authors examined participants' estimates of own and parental psychometric intelligence (IQ) and emotional intelligence (EI). The authors asked 224 participants (82 men, 138 women, 4 people who did not report their gender) to estimate their own and their parents' IQ and EI scores on a normal distribution ranging from 55 to 145 points. The authors hypothesized that men would give higher IQ but lower EI self-estimates than women and that participants, regardless of gender, would rate their fathers as higher on IQ but lower on EI than their mothers. The results confirmed the hypotheses, supporting the view that people perceive psychometric intelligence as a primarily masculine attribute in contrast with emotional intelligence, which they perceive as a primarily feminine attribute. The results also showed that the intensity of the stereotypical perception of EI as a feminine attribute diminished when the authors asked participants to estimate their scores on a range of specific EI facets instead of providing a direct overall self-estimate.


Dried Plasmodium falciparum-infected samples as positive controls for malaria rapid diagnostic tests.
Rapid diagnostic tests (RDTs) are central to fulfilling the WHO's recommendation for parasitologic confirmation of all suspected cases of malaria. RDT performance may be compromised when exposed to the high temperature conditions typical of most malaria endemic regions. However, a systematic method to monitor RDT quality and performance in endemic countries is lacking at the present time. Current methods to monitor RDT performance in the field include comparing results from RDTs to diagnoses made by light microscopy and observing health workers perform tests. These methods are not substitutes for direct quality control. In this study, the suitability of dried Plasmodium falciparum-infected blood as quality control samples for malaria RDTs was evaluated. Three cultured strains of P. falciparum at 200 and 2,000 parasites/μl were tested on 10 brands of RDT. After baseline testing to determine initial reactivity, aliquots of parasite-infected blood were air dried, stored at 35°C, room temperature (~25°C) or 4°C for one, four and 12 weeks and were then tested on the 10 RDTs after rehydration. Extended stability testing of dried blood stored at 4°C was done using P. falciparum strain 3D7 at 1,000 and 2,000 parasites/μl. All dried blood samples at 2,000 parasites/μl retained reactivity (100% sensitivity) at all three temperatures and time points for all nine RDT brands that detect histidine-rich protein-2 (HRP2). The dried blood samples with 200 parasites/μl were detected by six of the nine HRP2-based RDTs at all storage temperatures and time points. The sensitivity for two of the three remaining HRP2-based RDTs was 100% up to four weeks of storage at all temperatures but dropped to 87.5% at week 12. Of the four RDTs that detect plasmodium lactate dehydrogenase (pLDH) in a pan-specific manner, alone or in combination with HRP2, the detection of pLDH in samples with 2,000 parasites/μL was 100% for two RDTs and 80% for the other two RDTs. The mean level for detection of pLDH at 200 parasites/μl was low (29%), with a range of 0% to100%, which was partly attributable to weak initial baseline reactivity. Reactivity of dried 3D7 at 1,000 and 2,000 parasites/μl stored at 4°C was retained at 100% for up to 52 weeks for both HRP2 and pLDH. In the absence of native or recombinant positive control antigens, well-standardized P. falciparum-infected dried blood samples can be used as positive control samples for monitoring RDT performance, particularly with HRP2-detecting tests.


CD40: a growing cytoplasmic tale.
CD40, a member of the tumor necrosis factor (TNF) receptor family that is expressed on B cells, monocytes, dendritic cells, endothelial cells, and epithelial cells, as well as on B cell lymphomas and carcinomas, activates multiple signaling pathways. In B cells, the response to CD40 is complex and depends on the maturation status of the cell. It is well established that CD40 can promote cell survival through up-regulation of the expression of genes encoding antiapoptotic proteins. However, a new role for CD40 signaling is being recognized in promoting progression through the cell cycle. The roles of the phosphoinositide 3-kinase, mitogen-activated protein kinase, and nuclear factor kappaB pathways in mediating CD40 stimulation of the cell cycle are described.


Comparison of pre-analytical FFPE sample preparation methods and their impact on massively parallel sequencing in routine diagnostics.
Over the last years, massively parallel sequencing has rapidly evolved and has now transitioned into molecular pathology routine laboratories. It is an attractive platform for analysing multiple genes at the same time with very little input material. Therefore, the need for high quality DNA obtained from automated DNA extraction systems has increased, especially to those laboratories which are dealing with formalin-fixed paraffin-embedded (FFPE) material and high sample throughput. This study evaluated five automated FFPE DNA extraction systems as well as five DNA quantification systems using the three most common techniques, UV spectrophotometry, fluorescent dye-based quantification and quantitative PCR, on 26 FFPE tissue samples. Additionally, the effects on downstream applications were analysed to find the most suitable pre-analytical methods for massively parallel sequencing in routine diagnostics. The results revealed that the Maxwell 16 from Promega (Mannheim, Germany) seems to be the superior system for DNA extraction from FFPE material. The extracts had a 1.3-24.6-fold higher DNA concentration in comparison to the other extraction systems, a higher quality and were most suitable for downstream applications. The comparison of the five quantification methods showed intermethod variations but all methods could be used to estimate the right amount for PCR amplification and for massively parallel sequencing. Interestingly, the best results in massively parallel sequencing were obtained with a DNA input of 15 ng determined by the NanoDrop 2000c spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). No difference could be detected in mutation analysis based on the results of the quantification methods. These findings emphasise, that it is particularly important to choose the most reliable and constant DNA extraction system, especially when using small biopsies and low elution volumes, and that all common DNA quantification techniques can be used for downstream applications like massively parallel sequencing.


Effect of coronary occlusion during percutaneous transluminal angioplasty in humans on left ventricular chamber stiffness and regional diastolic pressure-radius relations.
The effect of repeated (3 to 10 second) and transient (15 to 75 second) abrupt coronary occlusion on the global and regional chamber stiffness was studied in nine patients undergoing angioplasty of a single proximal left anterior descending coronary artery stenosis. The left ventricular high fidelity pressure and volume relation was obtained before and after the procedure as well as during coronary occlusion, after 20 seconds (n = 9) and after 50 seconds (n = 5). During ischemia, there was an upward shift of the pressure-volume relation. The nonlinear simple elastic constant of chamber stiffness increased from 0.0273 +/- 0.017 before angioplasty (mean +/- SD) to 0.0621 +/- 0.026 after 20 seconds of occlusion (p less than 0.05) and 0.0605 +/- 0.015 after 50 seconds of occlusion (p less than 0.01). In five patients, the postangioplasty value remained higher than the control value, but at the group level the mean value (0.0529 +/- 0.037) was not statistically different. The regional stiffness was determined from the changes in the length of six segmental radii during diastole, from the lowest diastolic to the end-diastolic pressure. The regional constant of elastic stiffness was unaffected in the nonischemic zone. In the adjacent and ischemic zones, the regional stiffness was increased during occlusion (p less than 0.05). These regional abnormalities in diastolic function persisted at the time of postangioplasty measurements, 12 minutes after the end of the procedure. This suggests that recovery of normal diastolic function after repeated ischemic injuries is delayed after restoration of normal blood flow and systolic function.


Dietary fiber is associated with circulating concentrations of C-reactive protein in breast cancer survivors: the HEAL study.
Inflammation is a suspected risk factor for breast cancer and its subsequent prognosis. The extent to which dietary and lifestyle factors might influence inflammation is important to examine. Specifically, dietary fiber may reduce systemic inflammation, but this relationship has not been examined among breast cancer survivors. We examined associations between dietary fiber and serum concentrations of C-reactive protein (CRP) and serum amyloid A (SAA), among 698 female breast cancer survivors from the Health, Eating, Activity, and Lifestyle (HEAL) Study. Data are from interviews and clinical visits conducted 24-months post-study enrollment. Multivariate-adjusted linear regression estimated associations of total, soluble, and insoluble fiber with serum concentrations of CRP and SAA. Logistic regression estimated the odds of elevated CRP (defined as >3.0 mg/l) across tertiles of dietary fiber intake. Mean total dietary fiber intake was 13.9 ± 6.4 g/day. Mean CRP and SAA were 3.32 ± 3.66 and 7.73 ± 10.23 mg/l, respectively. We observed a multivariate-adjusted inverse association between total dietary fiber intake and CRP concentrations (β, -0.029; 95% CI, -0.049, -0.008). Results for insoluble fiber were similar (β, -0.039; 95% CI, -0.064, -0.013). Among survivors who consumed >15.5 g/day of insoluble dietary fiber, a 49% reduction in the likelihood of having elevated CRP concentrations (OR, 0.51; 95% CI, 0.27, 0.95) was observed compared to those who consumed <5.4 g/day (P = 0.053). Our results suggest that diets high in fiber may benefit breast cancer survivors via reductions in systemic inflammation; elevated inflammation may be prognostic for reduced survival.


Detection and characterization of anti-envoplakin linker autoantibodies in paraneoplastic pemphigus using specific bead-based assay.
Paraneoplastic pemphigus (PNP) is characterized by an autoantibody response directed against desmosomal antigens, desmogleins and plakin's proteins, i.e., periplakin and envoplakin. Notably, PNP antibodies were shown to recognize major epitopes located in the linker subdomain of human envoplakin and thus, may constitute a diagnostic marker of PNP. In this study, a recombinant envoplakin-linker subdomain (rENV-L) was produced and used to develop a specific bead-based assay to determine the prevalence and titers of anti-rENV-L antibodies in patients with different types of autoimmune bullous skin diseases. Sera from 33 PNP, 41 pemphigus vulgaris, 46 pemphigus foliaceus, 24 bullous pemphigoid and 74 normal subjects were analyzed by rENV-L bead-based assay: 23/33 (69.7%) PNP, 2/24 (8.3%) bullous pemphigoid and 4/74 (5.4%) healthy control sera showed IgG reactivity against rENV-L. For PNP, the sensitivity and specificity of the assay were 69.7% and 94.6%, respectively. Among the PNP sera reacting with envoplakin and periplakin by immunoblot analysis, 80.8% contained anti-rENV-L antibodies. Finally, we showed that an anti-rENV-L mAb that recognizes both envoplakin and periplakin gave the same fluorescence pattern on rat bladder sections than PNP sera. Thus, anti-rENV-L antibodies constitute a useful diagnostic marker of PNP and our bead-based assay, notably combined with other epidermal autoantigens, a useful tool to diagnose PNP.


Mitochondrial function after asphyxia in newborn lambs.
We examined mitochondrial oxidative function 5 minutes and 2 hours after a gradual asphyxial insult in newborn lambs. We subjected 16 ventilated newborn lambs to 75-90 minutes of hypoxia and hypercarbia that resulted in bradycardia and systemic hypotension over the final 15 minutes of the insult. At the end of asphyxia, the lambs were resuscitated and returned to control ventilator settings. Samples of brain were removed 5 minutes (n = 8) and 2 hours (n = 8) after asphyxia. Each group of eight lambs was subdivided into those less than 3 or greater than 3 days old to evaluate the effect of age on postasphyxia mitochondrial function. After classification into nonsynaptic and synaptic mitochondria, mitochondrial respiration (oxygen consumption) was measured using five different substrates. Data from asphyxiated lambs were compared with that from a control group of ventilated nonasphyxiated lambs (n = 8). In the lambs less than 3 days old, there was significant depression of mean +/- SEM nonsynaptic mitochondrial state 3 (adenosine diphosphate-dependent) respiration to 29.5 +/- 5.2% of control with four of the five substrates and of state 4 respiration to 33.7 +/- 0.9% of control with three of the five substrates 5 minutes after asphyxia. By 2 hours after asphyxia, mean +/- SEM nonsynaptic mitochondria state 3 respiration increased to 70.4 +/- 6.4% of control while state 4 respiration increased to 58.2 +/- 4.5% of control. In contrast, lambs greater than 3 days old exhibited no inhibition of nonsynaptic mitochondrial function after asphyxia.(ABSTRACT TRUNCATED AT 250 WORDS)


Enhanced efficacy of the CDNF/MANF family by combined intranigral overexpression in the 6-OHDA rat model of Parkinson's disease.
Cerebral Dopamine Neurotrophic Factor (CDNF) and Mesencephalic Astrocyte-derived Neurotrophic factor (MANF) are members of a recently discovered family of neurotrophic factors (NTFs). Here, we used intranigral or intrastriatal lentiviral vector-mediated expression to evaluate their efficacy at protecting dopaminergic function in the 6-OHDA model of Parkinson's disease (PD). In contrast to the well-studied Glial-Derived Neurotrophic Factor (GDNF), no beneficial effects were demonstrated by striatal overexpression of either protein. Interestingly, nigral overexpression of CDNF decreased amphetamine-induced rotations and increased tyroxine hydroxylase (TH) striatal fiber density but had no effect on numbers of TH(+) cells in the SN. Nigral MANF overexpression had no effect on amphetamine-induced rotations or TH striatal fiber density but resulted in a significant preservation of TH(+) cells. Combined nigral overexpression of both factors led to a robust reduction in amphetamine-induced rotations, greater increase in striatal TH-fiber density and significant protection of TH(+) cells in the SN. We conclude that nigral CDNF and MANF delivery is more efficacious than striatal delivery. This is also the first study to demonstrate that combined NTF can have synergistic effects that result in enhanced neuroprotection, suggesting that multiple NTF delivery may be more efficacious for the treatment of PD than the single NTF approaches attempted so far.


Evidence of recent interspecies horizontal gene transfer regarding nucleopolyhedrovirus infection of Spodoptera frugiperda.
Baculoviruses are insect-associated viruses carrying large, circular double-stranded-DNA genomes with significant biotechnological applications such as biological pest control, recombinant protein production, gene delivery in mammals and as a model of DNA genome evolution. These pathogens infect insects from the orders Lepidoptera, Hymenoptera and Diptera, and have high species diversity which is expressed in their diverse biological properties including morphology, virulence or pathogenicity. Spodoptera frugiperda (Lepidoptera: Noctuidae), the fall armyworm, represents a significant pest for agriculture in America; it is a host for baculoviruses such as the Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) (Colombia strain, genotype A) having been classified as a Group II alphabaculovirus making it a very attractive target for bioinsecticidal use. Genome analysis by pyrosequencing revealed that SfMNPV ColA has 145 ORFs, 2 of which were not present in the other sequenced genotypes of the virus (SfMNPV-NicB, SfMNPV-NicG, SfMNPV-19 and SfMNPV-3AP2). An in-depth bioinformatics study showed that ORF023 and ORF024 were acquired by a recent homologous recombination process between Spodoptera frugiperda and Spodoptera litura (the Oriental leafworm moth) nucleopolyhedroviruses. Auxiliary genes are numerous in the affected locus which has a homologous region (hr3), a repetitive sequence associated with genome replication which became lost in SfColA along with 1 ORF. Besides, the mRNAs associated with two acquired genes appeared in the virus' life-cycle during the larval stage. Predictive studies concerning the theoretical proteins identified that ORF023 protein would be a phosphatase involved in DNA repair and that the ORF024 protein would be a membrane polypeptide associated with cell transport. The SfColA genome was thus revealed to be a natural recombinant virus showing evidence of recent horizontal gene transfer between different baculovirus species occurring in nature. This feature could be the cause of its high insecticidal power and therefore SfColA becomes a great candidate for bioinsecticide formulations.


Multiplex cell microarrays for high-throughput screening.
Microarray technology was developed in the early 1990s to measure the transcription levels of thousands of genes in parallel. The basic premise of high-density arraying has since been expanded to create cell microarrays. Cells on chip are powerful experimental tools for high-throughput and multiplex screening of samples or cellular functions. Miniaturization increases assay throughput while reducing both reagent consumption and cell population heterogeneity effect, making these systems attractive for a wide range of assays, from drug discovery to toxicology, stem cell research and therapy. It is usual to functionalize the surface of a substrate to design cell microarrays. One form of cell microarrays, the transfected cell microarray, wherein plasmid DNA or siRNA spotted on the surface of a substrate is reverse-transfected locally into adherent cells, has become a standard tool for parallel cell-based analysis. With the advent of technology, cells can also be directly spotted onto functionalized surfaces using robotic fluid-dispensing devices or printed directly on bio-ink material. We are providing herein an overview of the latest developments in optical cell microarrays allowing high-throughput and high-content analysis.


Quantum chemical approach for condensed-phase thermochemistry: proposal of a harmonic solvation model.
We propose a novel quantum chemical method, called the harmonic solvation model (HSM), for calculating thermochemical parameters in the condensed phase, particularly in the liquid phase. The HSM represents translational and rotational motions of a solute as vibrations interacting with a cavity wall of solvent molecules. As examples, the HSM and the ideal-gas model (IGM) were used for the standard formation reaction of liquid water, combustion reactions of liquid formic acid, methanol, and ethanol, vapor-liquid equilibration of water and ethanol, and dissolution of gaseous CO2 in water. The numerical results confirmed the reliability and applicability of the HSM. In particular, the temperature dependence of the Gibbs energy of liquid molecules was accurately reproduced by the HSM; for example, the boiling point of water was reasonably determined using the HSM, whereas the conventional IGM treatment failed to obtain a crossing of the two Gibbs energy curves for gaseous and liquid water.


G2/M arrest and apoptosis of human colorectal cancer cells induced by water extract from residues of jelly fig achene.
Jelly fig (Ficus awkeotsang) achenes have been utilized to prepare a traditional drink in Taiwan. Herein, we evaluated the effect of water extract from jelly fig seed residues (WERJFA) on cancer cells. WERJFA could inhibit the growth of human colorectal cancer cells, COLO205 and HT29 in both dose- and time-dependent manners. The flow cytometric analysis with propidium iodide (PI) showed that WERJFA primarily arrested COLO205 and HT29 cells at the G2/M phase of cell cycle as the concentration reached to at least 0.5 mg/ml. WERJFA induced apoptosis of these two cell lines, as evidenced by annexin V-FITC/PI and 4', 6-diamidino-2-phenylindole (DAPI) staining using flow cytometry and confocal microscopy, respectively. Reactive oxygen species (ROS) production and the loss of mitochondrial membrane potential in WERJFA-treated cells were detected by flow cytometry with H2DCF-DA and 5,5', 6,6'-Tetrachloro-1, 1', 3,3'-tetraethylbenzimidazolocarbocyanine iodide (JC-1). Our results showed that WERJFA exerted anti-proliferative and apoptotic effects on colorectal cancer cells. WERJFA arrested cell cycle, and caused apoptotic death in these cancer cells possibly via mitochondrial pathway involved with exceeding ROS level.


Patterns of acrorenal malformation associations.
Limb and urinary tract defects have frequently been reported to occur together as components of a single acrorenal field defect or in many multiple malformation syndromes. However, the concordance of such anomalies has rarely been studied on a population basis or the relationships between specific limb and renal defects defined. This paper documents the patterns of acrorenal associations seen in over 1,500,000 infants born in Hungary in 1975-1984. In all, 1 in 1,800 infants had a limb deficiency and 9% of these (75 cases) had a urinary tract anomaly. Urinary tract anomalies were most commonly seen with radial ray defects, micromelia and amelia. The commonest recognized patterns were VACTERL association and the cloacal exstrophy and caudal regression sequences. Chromosomal and single gene defects also occurred. Numerical taxonomic techniques delineated six main clusters of patients. Important groupings included micromelia with renal agenesis, split hand/foot with hydronephrosis, and radial ray anomalies with VACTERL defects. The radial ray groups differed in the nature of the VACTERL anomalies seen and with respect to laterality, symmetry, and non-VACTERL anomalies. There was a strong association of bilateral limb defects with bilateral renal anomalies and unilateral with unilateral. Ipsilateral defects tended to occur in typical VACTERL cases, while contralateral defects tended to occur with additional non VACTERL midline anomalies. Although renal and limb anomalies are associated, in almost all cases malformations in other systems are also present. The precise nature of the malformation patterns seen appear to reflect differences in the nature and magnitude of the underlying dysmorphogenetic processes as well as the timing of their effects.


Traumatic brain injury registry in Taiwan.
This project was designed to examine the epidemiology of traumatic brain injury (TBI) in Taiwan. A total of 58,563 cases of TBI was collected from 114 hospitals in Taiwan during the period July 1, 1988-June 30, 1994. Traffic accident was the major cause of TBI (69.4%), followed by falls and assaults. Motorcyclists accounted for the vast majority of TBI cases among traffic accident victims (64.5%). The Glasgow Coma Scale was used in assessing the severity. 41,646 cases (79.5%) were considered mild, 4,637 cases (8.9%) moderate, and 6,078 cases (11.6%) severe. Skull x-ray showed fracture in 7,663 cases (14.6%). Intracranial hemorrhage was identified in 28.6% of patients receiving CT scanning. Craniotomy was performed in 5,226 cases (9%). The outcome of TBI was determined by the Glasgow Outcome Scale. Death occurred in 2,621 cases (5.4%), vegetative state in 429 cases (0.9%), severe disability in 1,293 cases (2.6%), moderate disability in 1,890 cases (3.9%), and good recovery in 42,596 cases (87.2%). The severity and outcome were worse than those of Western reports. In order to alleviate this problem, a helmet use persuasion program was conducted by the Police Department in Taipei City from January to June, 1994. Results of this program showed a significant reduction of TBI-related hospitalization, severity and fatality during this period of intervention. This study points out the seriousness of TBI in Taiwan and suggests some approaches and priorities for prevention.


[Diagnostic value of endobronchial ultrasound-guided transbronchial needle aspiration in stage Ⅰ and stage Ⅱ of sarcoidosis].
To evaluate the diagnostic value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in stage Ⅰ and stage Ⅱ of sarcoidosis. There were 55 patients of sarcoidosis selected from January 2012 to October 2014 in the First Affiliated Hospital of Soochow University. The diagnostic positive rate and the positive rate in stage Ⅰ and Ⅱ patients with sarcoidosis through EBUS-TBNA, conventional transbronchial needle aspiration (cTBNA), endobronchial biopsy (EBB) combined with transbronchial lung biopsy (TBLB) were calculated. The positive rate of single lymph node puncture and the positive rate of different size of lymph node were compared. The difference of the positive rate of lymph nodes in different stations was checked. By calculating the diagnostic yield of EBUS-TBNA in sarcoidosis patients, the practicality and safety of EBUS-TBNA in diagnosing stage Ⅰ or stage Ⅱ sarcoidosis was assessed. Among 55 patients, 46 patients obtained positive results through EBUS-TBNA or cTBNA. There were 18 patients who had only received EBUS-TBNA among a total of 55 patients diagnosed with sarcoidosis, positive results appeared in 17 cases, while cTBNA was 9/15. There were 20 cases diagnosed in which both had EBUS-TBNA and cTBNA. The diagnostic rate of cTBNA combined with EBB and TBLB was 25/26, while combined with EBUS-TBNA was 21/21. Totally 90 lymph nodes were punctured by EBUS-TBNA, in which 65 lymph nodes got the positive results (65/90) while 49 lymph nodes got the positive results by TBNA in 93 punctured lymph nodes (49/93). The diagnostic positive rate in the lymph nodes with a short diameter ≥2 cm was 33/37 by EBUS-TBNA, while it was 12/19 in those whose short diameter 1-<2 cm. By cTBNA, the diagnostic positive rate was 15/26 and 11/28. No serious adverse events occurred. The diagnosis of sarcoidosis in stage Ⅰ and stage Ⅱ by EBUS-TBNA is safe and effective, while choosing the most noticeable swelling lymph node to puncture is recommended. Combining the EBUS-TBNA with traditional bronchoscope technology can obtain a further higher diagnostic efficiency.


A pilot study on combination of cryosurgery and (125)iodine seed implantation for treatment of locally advanced pancreatic cancer.
To study the therapeutic value of combination of cryosurgery and (125)iodine seed implantation for locally advanced pancreatic cancer. Forty-nine patients with locally advanced pancreatic cancer (males 36, females 13), with a median age of 59 years, were enrolled in the study. Twelve patients had liver metastases. In all cases the tumors were considered unresectable after a comprehensive evaluation. Patients were treated with cryosurgery, which was performed intraoperatively or percutaneously under guidance of ultrasound and/or computed tomography (CT), and (125)iodine seed implantation, which was performed during cryosurgery or post-cryosurgery under guidance of ultrasound and/or CT. A few patients received regional celiac artery chemotherapy. Thirteen patients received intraoperative cryosurgery and 36 received percutaneous cryosurgery. Some patients underwent repeat cryosurgery. (125)Iodine seed implantation was performed during freezing procedure in 35 patients and 3-9 d after cryosurgery in 14 cases. Twenty patients, 10 of whom had hepatic metastases received regional chemotherapy. At 3 mo after therapy, CT was repeated to estimate tumor response to therapy. Most patients showed varying degrees of tumor necrosis. Complete response (CR) of tumor was seen in 20.4% patients, partial response (PR), in 38.8%, stable disease (SD), in 30.6%, and progressive disease (PD), in 10.2%. Adverse effects associated with cryosurgery included upper abdomen pain and increased serum amylase. Acute pancreatitis was seen in 6 patients one of whom developed severe pancreatitis. All adverse effects were controlled by medical management with no poor outcome. There was no therapy-related mortality. During a median follow-up of 18 mo (range of 5-40), the median survival was 16.2 mo, with 26 patients (53.1%) surviving for 12 mo or more. Overall, the 6-, 12-, 24- and 36-mo survival rates were 94.9%, 63.1%, 22.8% and 9.5%, respectively. Eight patients had survival of 24 mo or more. The patient with the longest survival (40 mo) is still living without evidence of tumor recurrence. Cryosurgery, which is far less invasive than conventional pancreatic resection, and is associated with a low rate of adverse effects, should be the treatment of choice for patients with locally advanced pancreatic cancer. (125)Iodine seed implantation can destroy the residual surviving cancer cells after cryosurgery. Hence, a combination of both modalities has a complementary effect.


On-line estimation of the dissolved zinc concentration during ZnS precipitation in a continuous stirred tank reactor (CSTR).
In this paper a method is presented to estimate the reaction term of zinc sulphide precipitation and the zinc concentration in a CSTR, using the read-out signal of a sulphide selective electrode. The reaction between zinc and sulphide is described by a non-linear model and therefore classical observer theory cannot be applied directly, as this theory was initially developed for linear systems. However, by linear reparametrization of this non-linear system, the linear observer theory can be applied in an effective way. This is illustrated by a zinc sulphide example using real data.


Evaluation of clinical experience using cell-based therapies in patients with spinal cord injury: a systematic review.
Using a systematic approach, the authors evaluated the current utilization, safety, and effectiveness of cellular therapies for traumatic spinal cord injuries (SCIs) in humans. A systematic search and critical review of the literature published through mid-January 2012 was performed. Articles included in the search were restricted to the English language, studies with at least 10 patients, and those analyzing cellular therapies for traumatic SCI. Citations were evaluated for relevance using a priori criteria, and those that met the inclusion criteria were critically reviewed. Each article was then designated a level of evidence that was developed by the Oxford Centre for Evidence-Based Medicine. The initial literature search identified 651 relevant articles, which decreased to 350 after excluding case reports and reviews. Evaluation of articles at the title/abstract level, and later at the full-text level, limited the final article set to 12 papers. The following cellular therapies employed in humans with SCI are reviewed: bone marrow mesenchymal and hematopoietic stem cells (8 studies), olfactory ensheathing cells (2 studies), Schwann cells (1 study), and fetal neurogenic tissue (1 study). Overall the quality of the literature was very low, with 3 Grade III levels of evidence and 9 Grade IV studies. Several different cellular-mediated strategies for adult SCI have been reported to be relatively safe with varying degrees of neurological recovery. However, the literature is of low quality and there is a need for improved preclinical studies and prospective, controlled clinical trials.


Enhanced transfer of a photocross-linking N-acetylglucosamine (GlcNAc) analog by an O-GlcNAc transferase mutant with converted substrate specificity.
O-Linked β-N-acetylglucosamine (O-GlcNAc) is a post-translational modification of proteins in multicellular organisms. O-GlcNAc modification is catalyzed by the O-GlcNAc transferase (OGT), which transfers N-acetylglucosamine (GlcNAc) from the nucleotide sugar donor UDP-GlcNAc to serine or threonine residues of protein substrates. Recently, we reported a novel metabolic labeling method to introduce the diazirine photocross-linking functional group onto O-GlcNAc residues in mammalian cells. In this method, cells are engineered to produce diazirine-modified UDP-GlcNAc (UDP-GlcNDAz), and the diazirine-modified GlcNAc analog (GlcNDAz) is transferred to substrate proteins by endogenous OGT, producing O-GlcNDAz. O-GlcNDAz-modified proteins can be covalently cross-linked to their binding partners, providing information about O-GlcNAc-dependent interactions. The utility of the method was demonstrated by cross-linking highly O-GlcNAc-modified nucleoporins to proteins involved in nuclear transport. For practical application of this method to a broader range of O-GlcNAc-modified proteins, efficient O-GlcNDAz production is critical. Here we examined the ability of OGT to transfer GlcNDAz and found that the wild-type enzyme (wtOGT) prefers the natural substrate, UDP-GlcNAc, over the unnatural UDP-GlcNDAz. This competition limits O-GlcNDAz production in cells and the extent of O-GlcNDAz-dependent cross-linking. Here we identified an OGT mutant, OGT(C917A), that efficiently transfers GlcNDAz and, surprisingly, has altered substrate specificity, preferring to transfer GlcNDAz rather than GlcNAc to protein substrates. We confirmed the reversed substrate preference by determining the Michaelis-Menten parameters describing the activity of wtOGT and OGT(C917A) with both UDP-GlcNAc and UDP-GlcNDAz. Use of OGT(C917A) enhances O-GlcNDAz production, yielding improved cross-linking of O-GlcNDAz-modified molecules both in vitro and in cells.


Caring for Critically Ill Children With Suspected or Proven Coronavirus Disease 2019 Infection: Recommendations by the Scientific Sections' Collaborative of the European Society of Pediatric and Neonatal Intensive Care.
In children, coronavirus disease 2019 is usually mild but can develop severe hypoxemic failure or a severe multisystem inflammatory syndrome, the latter considered to be a postinfectious syndrome, with cardiac involvement alone or together with a toxic shock like-presentation. Given the novelty of severe acute respiratory syndrome coronavirus 2, the causative agent of the recent coronavirus disease 2019 pandemic, little is known about the pathophysiology and phenotypic expressions of this new infectious disease nor the optimal treatment approach. From inception to July 10, 2020, repeated PubMed and open Web searches have been done by the scientific section collaborative group members of the European Society of Pediatric and Neonatal Intensive Care. There is little in the way of clinical research in children affected by coronavirus disease 2019, apart from descriptive data and epidemiology. Even though basic treatment and organ support considerations seem not to differ much from other critical illness, such as pediatric septic shock and multiple organ failure, seen in PICUs, some specific issues must be considered when caring for children with severe coronavirus disease 2019 disease. In this clinical guidance article, we review the current clinical knowledge of coronavirus disease 2019 disease in critically ill children and discuss some specific treatment concepts based mainly on expert opinion based on limited experience and the lack of any completed controlled trials in children at this time.


The Effect of Timing and Frequency of Push Notifications on Usage of a Smartphone-Based Stress Management Intervention: An Exploratory Trial.
Push notifications offer a promising strategy for enhancing engagement with smartphone-based health interventions. Intelligent sensor-driven machine learning models may improve the timeliness of notifications by adapting delivery to a user's current context (e.g. location). This exploratory mixed-methods study examined the potential impact of timing and frequency on notification response and usage of Healthy Mind, a smartphone-based stress management intervention. 77 participants were randomised to use one of three versions of Healthy Mind that provided: intelligent notifications; daily notifications within pre-defined time frames; or occasional notifications within pre-defined time frames. Notification response and Healthy Mind usage were automatically recorded. Telephone interviews explored participants' experiences of using Healthy Mind. Participants in the intelligent and daily conditions viewed (d = .47, .44 respectively) and actioned (d = .50, .43 respectively) more notifications compared to the occasional group. Notification group had no meaningful effects on percentage of notifications viewed or usage of Healthy Mind. No meaningful differences were indicated between the intelligent and non-intelligent groups. Our findings suggest that frequent notifications may encourage greater exposure to intervention content without deterring engagement, but adaptive tailoring of notification timing does not always enhance their use. Hypotheses generated from this study require testing in future work. ISRCTN67177737.


Alloy Microstructure Dictates Corrosion Modes in THA Modular Junctions.
Adverse local tissue reactions (ALTRs) triggered by corrosion products from modular taper junctions are a known cause of premature THA failure. CoCrMo devices are of particular concern because cobalt ions and chromium-orthophosphates were shown to be linked to ALTRs, even in metal-on-polyethylene THAs. The most common categories of CoCrMo alloy are cast and wrought alloy, which exhibit fundamental microstructural differences in terms of grain size and hard phases. The impact of implant alloy microstructure on the occurring modes of corrosion and subsequent metal ion release is not well understood. The purpose of this study was to determine whether (1) the microstructure of cast CoCrMo alloy varies broadly between manufacturers and can dictate specific corrosion modes; and whether (2) the microstructure of wrought CoCrMo alloy is more consistent between manufacturers and has low implications on the alloy's corrosion behavior. The alloy microstructure of four femoral-stem and three femoral-head designs from four manufacturers was metallographically and electrochemically characterized. Three stem designs were made from cast alloy; all three head designs and one stem design were made from wrought alloy. Alloy samples were sectioned from retrieved components and then polished and etched to visualize grain structure and hard phases such as carbides (eg, M23C6) or intermetallic phases (eg, σ phase). Potentiodynamic polarization (PDP) tests were conducted to determine the corrosion potential (Ecorr), corrosion current density (Icorr), and pitting potential (Epit) for each alloy. Four devices were tested within each group, and each measurement was repeated three times to ensure repeatable results. Differences in PDP metrics between manufacturers and between alloys with different hard phase contents were compared using one-way analysis of variance and independent-sample t-tests. Microstructural features such as twin boundaries and slip bands as well as corrosion damage features were viewed and qualitatively assessed in a scanning electron microscope. We found broad variability in implant alloy microstructure for both cast and wrought alloy between manufacturers, but also within the same implant design. In cast alloys, there was no difference in PDP metrics between manufacturers. However, coarse hard phases and clusters of hard phases (mainly intermetallic phases) were associated with severe phase boundary corrosion and pitting corrosion. Furthermore, cast alloys with hard phases had a lower Epit than those without (0.46 V, SD 0.042; 0.53 V, SD 0.03, respectively; p = 0.015). Wrought alloys exhibited either no hard phases or numerous carbides (M23C6). However, the corrosion behavior was mainly affected by lattice defects and banded structures indicative of segregations that appear to be introduced during bar stock manufacturing. Alloys with banding had a lower Ecorr (p = 0.008) and higher Icorr (p = 0.028) than alloys without banding (-0.76 V, SD 0.003; -0.73 V, SD 0.009; and 1.14 × 10-4 mA/cm2, SD 1.47 × 10-5; 5.2 × 10-5 mA/cm2, SD 2.57 × 10-5, respectively). Alloys with carbides had a slightly higher Ecorr (p = 0.046) than those without (-0.755 V, SD 0.005; -0.761 V, SD 0.004); however, alloys with carbides exhibited more severe corrosion damage as a result of phase boundary corrosion, hard phase detachment, and subsequent local crevice corrosion. The observed variability in CoCrMo alloy microstructure of both cast and wrought components in this study appears to be an important issue to address, perhaps through better standards, to minimize in vivo corrosion. The finding of the banded structures within wrought alloys is especially concerning because it unfavorably influences the corrosion behavior independent of the manufacturer. The findings suggest that a homogeneous alloy microstructure with a minimal hard phase fraction exhibits more favorable corrosion behavior within the in vivo environment of modular taper junctions, thus lowering metal ion release and subsequently the risk of ALTRs to corrosion products. Also, the question arises if hard phases fulfill a useful purpose in metal-on-polyethylene bearings, because they may come with a higher risk of phase boundary corrosion and pitting corrosion and the benefit they provide by adding strength is not needed (unlike in metal-on-metal bearings). Implant failure resulting from corrosion processes within modular junctions is a major concern in THA. Our results suggest that implant alloy microstructure is not sufficiently standardized and may also dictate specific corrosion modes and subsequent metal ion release.


Systems level analysis of the Chlamydomonas reinhardtii metabolic network reveals variability in evolutionary co-conservation.
Metabolic networks, which are mathematical representations of organismal metabolism, are reconstructed to provide computational platforms to guide metabolic engineering experiments and explore fundamental questions on metabolism. Systems level analyses, such as interrogation of phylogenetic relationships within the network, can provide further guidance on the modification of metabolic circuitries. Chlamydomonas reinhardtii, a biofuel relevant green alga that has retained key genes with plant, animal, and protist affinities, serves as an ideal model organism to investigate the interplay between gene function and phylogenetic affinities at multiple organizational levels. Here, using detailed topological and functional analyses, coupled with transcriptomics studies on a metabolic network that we have reconstructed for C. reinhardtii, we show that network connectivity has a significant concordance with the co-conservation of genes; however, a distinction between topological and functional relationships is observable within the network. Dynamic and static modes of co-conservation were defined and observed in a subset of gene-pairs across the network topologically. In contrast, genes with predicted synthetic interactions, or genes involved in coupled reactions, show significant enrichment for both shorter and longer phylogenetic distances. Based on our results, we propose that the metabolic network of C. reinhardtii is assembled with an architecture to minimize phylogenetic profile distances topologically, while it includes an expansion of such distances for functionally interacting genes. This arrangement may increase the robustness of C. reinhardtii's network in dealing with varied environmental challenges that the species may face. The defined evolutionary constraints within the network, which identify important pairings of genes in metabolism, may offer guidance on synthetic biology approaches to optimize the production of desirable metabolites.