Genetic variation in the neuropeptide Y gene promoter is associated with increased risk of tobacco smoking.
Neuropeptide Y (NPY) is a strong candidate gene regarding the pathophysiology of tobacco dependence. It has been associated with various addictive and psychiatric disorders, and closely interacts with the brain reward system. The aim of the present study was to test for association between a functional genetic variant in the NP-Y promoter gene (SNP rs16147) and tobacco smoking. In a population-based case-control multicenter study designed for tobacco addiction research, a total of 550 Caucasian current smokers, and 544 never-smokers were genotyped for SNP rs16147 and behaviorally characterized with the State-Trait Anxiety Inventory (STAI). Subjects with TT genotype of the SNP rs16147 were significantly more frequently smokers than never-smokers (p = 0.046). In addition, TT genotype exhibited increased state anxiety scores compared to carriers of the C allele (p = 0.037). Our results provide evidence for an involvement of the functionally relevant SNP rs16147 in the pathophysiology of tobacco dependence. Further studies are needed to confirm our findings.


Excimer laser phototherapeutic keratectomy: indications, results and its role in the Indian scenario.
To report indications, technique, and results of excimer phototherapeutic keratectomy (PTK), and describe possible reasons for the small numbers of such procedures performed in a referral institute in India. Retrospective review of case records of 10 patients (11 eyes) who underwent excimer PTK at our institute between February 1994 and September 1997. Corneal scars were the most common indication for treatment. Best-corrected visual acuity (BCVA) improved in 6 eyes (mean: 2 lines of Snellen acuity). All eyes had BCVA > or = 6/12 after treatment. None of the patients experienced loss of BCVA after treatment. Unaided visual acuity improved in 3 eyes and decreased in 2 eyes. Change in spherical equivalent refraction > or = 1 diopter occurred in 77.8% of eyes after treatment. Treating central corneal scars resulted in a significant hyperopic shift in refraction. Excimer PTK is a safe and effective procedure for the treatment of superficial corneal opacities. Post-treatment ametropia may require further correction with optical aids. Inappropriate referrals, deep corneal scars, and cost of the procedure could have contributed to the small numbers of PTK performed at our institute. Improved understanding of procedural strengths and limitations could lead to increased use of this procedure, with satisfying results in selected patients.


Fabrication and Characterization of Nanocarbon-Based Nanofluids by Using an Oxygen-Acetylene Flame Synthesis System.
In this study, an oxygen-acetylene flame synthesis system was developed to fabricate nanocarbon-based nanofluids (NCBNFs) through a one-step synthesis method. Measured in liters per minute (LPM), the flame's fuel flows combined oxygen and acetylene at four ratios: 1.5/2.5 (P1), 1.0/2.5 (P2), 0.5/2.5 (P3), and 0/2.5 (P4). The flow rate of cooling water (base fluid) was fixed at 1.2 LPM to produce different nanocarbon-based materials (NCBMs) and various concentrations of NCBNFs. Tests and analyses were conducted for determining the morphology of NCBMs, NCBM material, optical characteristics, the production rate, suspension performance, average particle size, zeta potential, and other relevant basic characteristics of NCBNFs to understand the characteristics and materials of NCBNFs produced through different process parameters (P1-P4). The results revealed that the NCBMs mainly had flaky and spherical morphologies and the diameters of the spherical NCBMs measured approximately 20-30 nm. X-ray diffraction and Raman spectroscopy revealed that the NCBMs contained graphene oxide (GO) and amorphous carbon (AC) when the oxygen flow rate was lower than 1.0 LPM. In addition, the NCBMs contained reduced GO, crystalline graphite (graphite-2H), and AC when the oxygen flow rate was higher than 1.0 LPM. The process parameters of P1, P2, P3, and P4 resulted in NCBMs produced at concentrations of 0.010, 0.013, 0.040, and 0.023 wt%, respectively, in NCBNFs. All the NCBNFs exhibited non-Newtonian and shear-thinning rheological properties. The P4 ratio showed the highest enhancement rate of thermal conductivity for NCBNFs, at a rate 4.85 % higher than that of water.


The influence of active coping and perceived stress on health disparities in a multi-ethnic low income sample.
Extensive research has shown that ethnic health disparities are prevalent and many psychological and social factors influence health disparities. Understanding what factors influence health disparities and how to eliminate health disparities has become a major research objective. The purpose of this study was to examine the impact of coping style, stress, socioeconomic status (SES), and discrimination on health disparities in a large urban multi-ethnic sample. Data from 894 participants were collected via telephone interviews. Independent variables included: coping style, SES, sex, perceived stress, and perceived discrimination. Dependent variables included self-rated general and oral health status. Data analysis included multiple linear regression modeling. Coping style was related to oral health for Blacks (B = .23, p < .05) and for Whites there was a significant interaction (B = -.59, p < .05) between coping style and SES for oral health. For Blacks, active coping was associated with better self-reported health. For Whites, low active coping coupled with low SES was significantly associated with worse oral health. Coping style was not significantly related to general health. Higher perceived stress was a significant correlate of poorer general health for all ethnoracial groups and poorer oral health for Hispanics and Blacks. SES was directly related to general health for Hispanics (.B = .27, p < .05) and Whites (B = .23, p < .05) but this relationship was mediated by perceived stress. Our results indicate that perceived stress is a critical component in understanding health outcomes for all ethnoracial groups. While SES related significantly to general health for Whites and Hispanics, this relationship was mediated by perceived stress. Active coping was associated only with oral health.


Hydrolysis of magnetic amylase-imprinted poly(ethylene-co-vinyl alcohol) composite nanoparticles.
Molecularly imprinted polymers (MIPs) have frequently been employed as recognition elements in sensing applications, or for the controlled delivery of small molecule drugs. An equally important but less well studied application is the use of MIPs in the binding and immobilization of active enzymes. In this study, magnetic MIPs (MMIPs) recognizing the enzyme amylase were prepared using phase inversion of poly(ethylene-co-vinyl alcohol) (EVAL) solutions with 27-44 mol % ethylene in the presence of amylase. The size distribution, specific surface area, magnetization, and composition were characterized by dynamic light scattering (DLS), Brunauer-Emmett-Teller (BET) analysis, superconducting quantum interference devices (SQUID), and X-ray diffraction (XRD), respectively. The mean size of MMIPs was ~100 nm and the magnetization was 14.8 emu/g. The activities of both bound template and rebound enzyme was established by measuring glucose production via starch hydrolysis, at different temperatures, for MIPs with different compositions (wt % EVALs and mol % ethylene). The highest hydrolysis activity of MMIPs (obtained with 32 mol % ethylene) was found to be 1545.2 U/g. Finally, compared to the conventional catalysis process, MMIPs have the advantages of high surface area, suspension, easy removal from reaction, and rapid reload of enzyme. The good activity of amylase MMIPs persists after 50 cycles of starch hydrolysis.


WITHDRAWN: Ozone therapy for the treatment of dental caries.
Dental caries is a bacterially mediated disease characterised by demineralisation of the tooth surface, which may lead to cavitation, discomfort, pain and eventual tooth loss. Ozone is toxic to certain bacteria in vitro and it has been suggested that delivering ozone into a carious lesion might reduce the number of cariogenic bacteria. This possibly could arrest the progress of the lesion and may, in the presence of fluoride, perhaps allow remineralisation to occur. This may in turn delay or prevent the need for traditional dental conservation by 'drilling and filling'. To assess whether ozone is effective in arresting or reversing the progression of dental caries. We searched the Cochrane Oral Health Group's Trials Register (to 7 November 2003); Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2003, Issue 3); MEDLINE and PREMEDLINE (OVID) (1966 to November 2003); EMBASE (OVID) (1980 to November 2003); CINAHL (OVID) (1982 to November 2003); AMED (OVID) (1985 to November 2003). Quintessence was handsearched through 2002 and KaVo were contacted as manufacturers of the HealOzone apparatus for any additional published or unpublished trials. Inclusion was assessed independently by at least two reviewers. Trials were only included if they met the following criteria: randomisation in a controlled trial; single surface in vivo carious lesion accessible to ozone application; clear allocation concealment; ozone application to the lesions in the intervention group; no such application of ozone in the control group; outcomes measured after at least 6 months. Reviewers independently extracted information in duplicate. A paucity of comparable data did not allow meta-analytic pooling of the included studies. Three trials were included, with a combined total of 432 randomised lesions (137 participants). Forty-two conference papers, abstracts and posters were excluded (from an unknown number of studies). The risk of bias in all studies appeared high. The analyses of all three studies were conducted at the level of the lesion, which is not independent of the person, for this reason pooling of data was not appropriate or attempted. Individual studies showed inconsistent effects of ozone on caries, across different measures of caries progression or regression. Few secondary outcomes were reported, but one trial reported an absence of adverse events. Given the high risk of bias in the available studies and lack of consistency between different outcome measures, there is no reliable evidence that application of ozone gas to the surface of decayed teeth stops or reverses the decay process. There is a fundamental need for more evidence of appropriate rigour and quality before the use of ozone can be accepted into mainstream primary dental care or can be considered a viable alternative to current methods for the management and treatment of dental caries.


Benzothiazole- and benzoxazole-substituted pyridine-2-carboxylates as efficient sensitizers of europium luminescence.
A facile synthesis of benzothiazole- and benzoxazole-substituted pyridine-2-carboxylic acids has been developed. These ligands form mononuclear nine-coordinate complexes [Ln(kappa(3)-ligand)(2)(kappa(1)-ligand)(H(2)O)(2)] with light and heavy trivalent lanthanides, as established from the X-ray analysis of 11 complexes. A crystal structure of a minor product, the anhydrous nine-coordinate complex [Eu(kappa(3)-L)(3)], has also been determined. Photophysical studies of gadolinium chelates indicate that the triplet states of the new ligands are located at 20400-21400 cm(-1). The ligands are good sensitizers of the europium luminescence with ligand-to-metal energy transfer efficiency in the range 60-100%. The overall quantum yields of the europium emission are substantial, 12-14% in the solid state, and increase to 29-39% upon replacement of two metal-coordinated water molecules with dimethylsulfoxide in solution. The luminescence of near-infrared emitting lanthanides is also sensitized, but quantum yields are much smaller, reaching 0.17% for neodymium and 1.25% for ytterbium in DMSO, while energy transfer efficiencies for these two ions are below 50%.


Selectivity and localization of lysozyme uptake in contemporary hydrogel contact lens materials.
The purpose of this study was to investigate the early and selective uptake of lysozyme and the location of deposited lysozyme on contemporary hydrogel contact lens (CL) materials after exposure to an artificial tear solution (ATS) for 16 h. Seven different hydrogel CL materials [polymacon, omafilcon A, nelfilcon A, nesofilcon A, ocufilcon B, etafilcon A (Acuvue Moist), and etafilcon A (Acuvue Define)] were incubated in an ATS for various times. Total protein deposition was determined using a modified Bradford technique. Lysozyme, lactoferrin, and albumin deposition on CLs were determined using 125I-radiolabeling method. A confocal laser scanning microscopy (CLSM) technique was utilized to map the location of lysozyme uptake in an asymmetric environment. All lens materials had significant amounts of lysozyme after 1 min of exposure to ATS. After 16 h of incubation, higher levels of total protein deposited on the two etafilcon A-based lenses (Moist and Define), followed by ocufilcon B and both were significantly higher than all other CLs tested (p = 0.0001). The two etafilcon A materials (Moist and Define) also deposited the highest amounts of lysozyme (514.8 ± 28.4 and 527.1 ± 14.7 μg/lens respectively) when compared to other test CLs (p = 0.0001). The CLSM technique revealed that the non-ionic CLs tended to have symmetric distribution of lysozyme throughout the lens materials, while the ionic CLs had an asymmetric distribution, with the highest concentration of lysozyme on and near the exposed surface. The quantity and nature of proteins deposited on CLs varies, depending upon the chemical composition of the material. Among the various lenses tested, etafilcon A deposited the highest amount of total protein, most of it represented by lysozyme, which was largely located near the surface of the lens.


Effect of dietary citric acid on the performance and mineral metabolism of broiler.
The objective of this study was to investigate the effect of dietary citric acid (CA) on the performance and mineral metabolism of broiler chicks. A total of 1720 Ross PM3 broiler chicks (days old) were randomly assigned to four groups (430 in each) and reared for a period of 35 days. The diets of groups 1, 2, 3 and 4 were supplemented with 0%, 0.25%, 0.75% or 1.25% CA by weight respectively. Feed and faeces samples were collected weekly and analysed for acid insoluble ash, calcium (Ca), phosphorus (P) and magnesium (Mg). The pH was measured in feed and faeces. At the age of 28 days, 10 birds from each group were slaughtered; tibiae were collected from each bird for the determination of bone mineral density, total ash, Ca, P, Mg and bone-breaking strength, and blood was collected for the measurement of osteocalcin, serum CrossLaps(®), Ca, P, Mg and 1,25(OH)(2)Vit-D in serum. After finishing the trial on day 37, all chicks were slaughtered by using the approved procedure. Birds that were fed CA diets were heavier (average body weights of 2030, 2079 and 2086 g in the 0.25%, 0.75% and 1.25% CA groups, respectively, relative to the control birds (1986 g). Feed conversion efficiency (weight gain in g per kg of feed intake) was also higher in birds of the CA-fed groups (582, 595 and 587 g/kg feed intake for 0.25%, 0.75% and 1.25% CA respectively), relative to the control birds (565 g/kg feed intake). The digestibility of Ca, P and Mg increased in the CA-fed groups, especially for the diets supplemented with 0.25% and 0.75% CA. Support for finding was also indicated in the results of the analysis of the tibia. At slaughter, the birds had higher carcass weights and higher graded carcasses in the groups that were fed the CA diets. The estimated profit margin was highest for birds fed the diet containing 0.25% CA. Birds of the 0.75% CA group were found to have the second highest estimated profit margin. Addition of CA up to a level of 1.25% of the diet increased performance, feed conversion efficiency, carcass weight and carcass quality, but only in numerical terms. The addition of CA up to 0.75% significantly increased the digestibility of macro minerals, bone ash content, bone mineral density and bone strength of the broiler chicks. It may, therefore, be concluded that the addition of 0.75% CA in a standard diet is suitable for growth, carcass traits, macromineral digestibility and bone mineral density of broiler chicks.


Apolipoprotein(B) identifies dyslipidemic phenotypes associated with cardiovascular risk in normocholesterolemic type 2 diabetic patients.
Apolipoprotein(B) [apo(B)] reflects the total mass of atherogenic particles (VLDL, IDL, and LDL), and its increase is associated with cardiovascular disease independently of LDL cholesterol (LDLc) levels. Apo(B) determination has been recently standardized, but attention to regional reference limits is advisable. Our aim was to analyze the frequency of dyslipidemic phenotypes, including those dependent on increased apo(B) in normocholesterolemic type 2 diabetic patients. A total of 100 consecutively seen type 2 diabetic patients (63 men, 37 women; aged 59 +/- 11 years) were included, after excluding those on lipid-lowering therapy. Apo(B) cutoff (1.1 g/l) was obtained from a group of normolipidemic (47 men, 21 women) control subjects, and LDLc, triglycerides, and HDL cholesterol (HDLc) cutoff points were those from the National Cholesterol Education Program guidelines. LDLc levels were obtained by ultracentrifugation if triglyceride levels were > 3.45 mmol/l; otherwise, they were calculated (Friedwald). Apo(B) levels were measured by immunoturbidimetry. Normocholesterolemia (LDLc < 4.13 mmol/l) appeared in 75 of the 100 patients, of whom 55 were normo- and 20 hypertriglyceridemic. Hyperapolipoprotein(B) [hyperapo(B)] was the most frequent lipid disorder, present in 34 (45%) of the normocholesterolemic patients (22 normo- and 12 hypertriglyceridemic). Low HDLc levels were more prevalent (53%) in patients with hyperapo(B) than in the rest (24%). Hyperapo(B) was found in almost half of the normocholesterolemic type 2 diabetic patients and was frequently associated with low HDLc levels and hypertriglyceridemia. Thus, given its independent association with cardiovascular disease and that it identifies high-risk phenotypes in normocholesterolemic diabetic patients apo(B) should be used to evaluate the lipidic pattern of these patients.


Predictors of difficult videolaryngoscopy with GlideScope® or C-MAC® with D-blade: secondary analysis from a large comparative videolaryngoscopy trial.
Tracheal intubation using acute-angle videolaryngoscopy achieves high success rates, but is not without difficulty. We aimed to determine predictors of 'difficult videolaryngoscopy'. We performed a secondary analysis of a data set (n=1100) gathered from a multicentre prospective randomized controlled trial of patients for whom difficult direct laryngoscopy was anticipated and who were intubated with one of two videolaryngoscopy devices (GlideScope(®) or C-MAC(®) with D-blade). 'Difficult videolaryngoscopy' was defined as 'first intubation time >60 s' or 'first attempt intubation failure'. A multivariate logistic regression model along with stepwise model selection techniques was performed to determine independent predictors of difficult videolaryngoscopy. Of 1100 patients, 301 were identified as difficult videolaryngoscopies. By univariate analysis, head and neck position, provider, type of surgery, and mouth opening were associated with difficult videolaryngoscopy (P<0.05). According to the multivariate logistic regression model, characteristics associated with greater risk for difficult videolaryngoscopy were as follows: (i) head and neck position of 'supine sniffing' vs 'supine neutral' {odds ratio (OR) 1.63, 95% confidence interval (CI) [1.14, 2.31]}; (ii) undergoing otolaryngologic or cardiac surgery vs general surgery (OR 1.89, 95% CI [1.19, 3.01] and OR 6.13, 95% CI [1.85, 20.37], respectively); (iii) intubation performed by an attending anaesthestist vs a supervised resident (OR 1.83, 95% CI [1.14, 2.92]); and (iv) small mouth opening (OR 1.18, 95% CI [1.02, 1.36]). This secondary analysis of an existing data set indicates four covariates associated with difficult acute-angle videolaryngoscopy, of which patient position and provider level are modifiable.


Quantitative imaging of ischemic stroke through thinned skull in mice with Multi Exposure Speckle Imaging.
Laser Speckle Contrast Imaging (LSCI) has become a widely used technique to image cerebral blood flow in vivo. However, the quantitative accuracy of blood flow changes measured through the thin skull has not been investigated thoroughly. We recently developed a new Multi Exposure Speckle Imaging (MESI) technique to image blood flow while accounting for the effect of scattering from static tissue elements. In this paper we present the first in vivo demonstration of the MESI technique. The MESI technique was used to image the blood flow changes in a mouse cortex following photothrombotic occlusion of the middle cerebral artery. The Multi Exposure Speckle Imaging technique was found to accurately estimate flow changes due to ischemia in mice brains in vivo. These estimates of these flow changes were found to be unaffected by scattering from thinned skull.


Cerebellar output controls generalized spike-and-wave discharge occurrence.
Disrupting thalamocortical activity patterns has proven to be a promising approach to stop generalized spike-and-wave discharges (GSWDs) characteristic of absence seizures. Here, we investigated to what extent modulation of neuronal firing in cerebellar nuclei (CN), which are anatomically in an advantageous position to disrupt cortical oscillations through their innervation of a wide variety of thalamic nuclei, is effective in controlling absence seizures. Two unrelated mouse models of generalized absence seizures were used: the natural mutant tottering, which is characterized by a missense mutation in Cacna1a, and inbred C3H/HeOuJ. While simultaneously recording single CN neuron activity and electrocorticogram in awake animals, we investigated to what extent pharmacologically increased or decreased CN neuron activity could modulate GSWD occurrence as well as short-lasting, on-demand CN stimulation could disrupt epileptic seizures. We found that a subset of CN neurons show phase-locked oscillatory firing during GSWDs and that manipulating this activity modulates GSWD occurrence. Inhibiting CN neuron action potential firing by local application of the γ-aminobutyric acid type A (GABA-A) agonist muscimol increased GSWD occurrence up to 37-fold, whereas increasing the frequency and regularity of CN neuron firing with the use of GABA-A antagonist gabazine decimated its occurrence. A single short-lasting (30-300 milliseconds) optogenetic stimulation of CN neuron activity abruptly stopped GSWDs, even when applied unilaterally. Using a closed-loop system, GSWDs were detected and stopped within 500 milliseconds. CN neurons are potent modulators of pathological oscillations in thalamocortical network activity during absence seizures, and their potential therapeutic benefit for controlling other types of generalized epilepsies should be evaluated.


K-space polarimetry of bullseye plasmon antennas.
Surface plasmon resonators can drastically redistribute incident light over different output wave vectors and polarizations. This can lead for instance to sub-diffraction sized nanoapertures in metal films that beam and to nanoparticle antennas that enable efficient conversion of photons between spatial modes, or helicity channels. We present a polarimetric Fourier microscope as a new experimental tool to completely characterize the angle-dependent polarization-resolved scattering of single nanostructures. Polarimetry allows determining the full Stokes parameters from just six Fourier images. The degree of polarization and the polarization ellipse are measured for each scattering direction collected by a high NA objective. We showcase the method on plasmonic bullseye antennas in a metal film, which are known to beam light efficiently. We find rich results for the polarization state of the beamed light, including complete conversion of input polarization from linear to circular and from one helicity to another. In addition to uncovering new physics for plasmonic groove antennas, the described technique projects to have a large impact in nanophotonics, in particular towards the investigation of a broad range of phenomena ranging from photon spin Hall effects, polarization to orbital angular momentum transfer and design of plasmon antennas.


Integrative and perturbation-based analysis of the transcriptional dynamics of TGFβ/BMP system components in transition from embryonic stem cells to neural progenitors.
Cooperative actions of extrinsic signals and cell-intrinsic transcription factors alter gene regulatory networks enabling cells to respond appropriately to environmental cues. Signaling by transforming growth factor type β (TGFβ) family ligands (eg, bone morphogenetic proteins [BMPs] and Activin/Nodal) exerts cell-type specific and context-dependent transcriptional changes, thereby steering cellular transitions throughout embryogenesis. Little is known about coordinated regulation and transcriptional interplay of the TGFβ system. To understand intrafamily transcriptional regulation as part of this system's actions during development, we selected 95 of its components and investigated their mRNA-expression dynamics, gene-gene interactions, and single-cell expression heterogeneity in mouse embryonic stem cells transiting to neural progenitors. Interrogation at 24 hour intervals identified four types of temporal gene transcription profiles that capture all stages, that is, pluripotency, epiblast formation, and neural commitment. Then, between each stage we performed esiRNA-based perturbation of each individual component and documented the effect on steady-state mRNA levels of the remaining 94 components. This exposed an intricate system of multilevel regulation whereby the majority of gene-gene interactions display a marked cell-stage specific behavior. Furthermore, single-cell RNA-profiling at individual stages demonstrated the presence of detailed co-expression modules and subpopulations showing stable co-expression modules such as that of the core pluripotency genes at all stages. Our combinatorial experimental approach demonstrates how intrinsically complex transcriptional regulation within a given pathway is during cell fate/state transitions.


Ascertainment bias and the pattern of nucleotide diversity at the human ALDH2 locus in a Japanese population.
Many East Asian human populations harbor a high-frequency deficiency allele for the aldehyde dehydrogenase 2 (ALDH2) enzyme, a critical protein involved in the metabolism of ethanol. Here we use resequencing and long-range SNP haplotype data from a Japanese sample to test whether patterns of nucleotide diversity and linkage disequilibrium at this locus are compatible with a standard neutral model of evolution. Examination of the pattern of polymorphism at a locus such as this, where the frequency of a common allele is known a priori, introduces an ascertainment bias that must be corrected for in analyses of the frequency spectrum of polymorphisms. We apply a flexible and generally applicable simulation approach to correct for this bias in our ALDH2 data and, also, to explore the effect of bias on the commonly used summary statistics Tajima's D, Fu and Li's D, and Fay and Wu's H. Our study finds no evidence that the pattern of genetic variation at ALDH2 differs from that expected under a standard neutral model. However, our general examination of ascertainment bias indicates that a priori knowledge of segregating alleles greatly affects the expected distributions of summary statistics. Under many parameter combinations we find that ascertainment bias introduces an elevated rate of false positives when summary statistics are used to test for deviations from a standard neutral model. However, we also show that over a wide range of conditions the power of all summary statistics can be greatly increased by incorporating prior knowledge of segregating alleles.


Precision and accuracy of ground reaction force normalization in a heterogeneous population of dogs.
To determine if currently used ground reaction force (GRF) normalization methods are accurate and precise enough to be used on a single-limb basis. Prospective clinical trial. Clinically normal (n = 69) dogs and 40 dogs with unilateral ruptured cranial cruciate ligaments (CCL). Pelvic limb GRFs of orthopedically normal dogs and those with unilateral ruptured CCL were collected. Normalization methods included none, body weight (BW), withers height (WH), WH and relative velocity (WH*F) and principal component 1 (PC1). Normalization methods were evaluated both by individual GRFs and additively. Binary logistic regression was performed for all normalization methods; sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) calculated. Stepwise backward logistic regression was used; significant values were retained in the final model. P < .05 was significant. Normalization of contact time (CT) by BW uniformly increased sensitivity, specificity, PPV, NPV, and accuracy. SI was the most accurate at both the walk and trot (accuracy 80-96%). Normalization by BW, WH, and WH*F all achieved similar results. When normalized GRFs were added, the accuracy increased only at the walk. CT should be normalized to BW. SIs remain the gold standard, if SIs cannot be used, combining GRFs normalized to BW will result in high precision (80%) and high accuracy (89.5%) at the walk. At the trot, normalization by BW, WH and WH*F results in consistent results for the individual GRFs, though not all accuracies are >80%.


Extensive rewiring of the EGFR network in colorectal cancer cells expressing transforming levels of KRASG13D.
Protein-protein-interaction networks (PPINs) organize fundamental biological processes, but how oncogenic mutations impact these interactions and their functions at a network-level scale is poorly understood. Here, we analyze how a common oncogenic KRAS mutation (KRASG13D) affects PPIN structure and function of the Epidermal Growth Factor Receptor (EGFR) network in colorectal cancer (CRC) cells. Mapping >6000 PPIs shows that this network is extensively rewired in cells expressing transforming levels of KRASG13D (mtKRAS). The factors driving PPIN rewiring are multifactorial including changes in protein expression and phosphorylation. Mathematical modelling also suggests that the binding dynamics of low and high affinity KRAS interactors contribute to rewiring. PPIN rewiring substantially alters the composition of protein complexes, signal flow, transcriptional regulation, and cellular phenotype. These changes are validated by targeted and global experimental analysis. Importantly, genetic alterations in the most extensively rewired PPIN nodes occur frequently in CRC and are prognostic of poor patient outcomes.


PIASy represses CCAAT/enhancer-binding protein delta (C/EBPdelta) transcriptional activity by sequestering C/EBPdelta to the nuclear periphery.
CCAAT/enhancer binding proteindelta (C/EBPdelta) plays a key role in mammary epithelial cell G(0) growth arrest, and "loss of function" alterations in C/EBPdelta have been reported in breast cancer and acute myeloid leukemia. C/EBPdelta is regulated at the transcriptional, post-transcriptional, and post-translational levels, suggesting tight control of C/EBPdelta content and function. Protein inhibitors of activated STATs (PIASs) regulate a growing number of transcription factors, including C/EBPs. HC11 nontransformed mammary epithelial cells express PIAS3, PIASxbeta, and PIASy, and all three PIAS family members repress C/EBPdelta transcriptional activity. PIASy is the most potent, however, repressing C/EBPdelta transcriptional activity by >80%. PIASy repression of C/EBPdelta transcriptional activity is dependent upon interaction between the highly conserved PIASy N-terminal nuclear matrix binding domain (SAPD) and the C/EBPdelta transactivation domain (TAD). PIASy repression of C/EBPdelta transcriptional activity is independent of histone deacetylase activity, PIASy E3 SUMO ligase activity, and C/EBPdelta sumoylation status. PIASy expression is associated with C/EBPdelta translocation from nuclear foci, where C/EBPdelta co-localizes with p300, to the nuclear periphery. PIASy-mediated translocation of C/EBPdelta is dependent upon the PIASy SAPD and C/EBPdelta TAD. PIASy reduces the expression of C/EBPdelta adhesion-related target genes and enhances repopulation of open areas within a cell monolayer in the in vitro "scratch" assay. These results demonstrate that PIASy represses C/EBPdelta by a mechanism that requires interaction between the PIASy SAPD and C/EBPdelta TAD and does not require PIASy SUMO ligase activity or C/EBPdelta sumoylation. PIASy alters C/EBPdelta nuclear localization, reduces C/EBPdelta transcriptional activity, and enhances cell proliferation/migration.


Higher cannabidiol plasma levels are associated with better seizure response following treatment with a pharmaceutical grade cannabidiol.
The objective of this study was to determine the relationship between cannabidiol (CBD) dose, CBD plasma level, and seizure control in a large open-label single-center study. All participants with treatment-refractory epilepsy participating in our expanded access program (EAP) were approached for participation. Highly purified grade CBD (Epidiolex®) dosing was weight-based and could be increased every 2 weeks by 5 mg/kg/day up to a maximum dosage of 50 mg/kg/day depending on tolerance and seizure control. Seizure counts were obtained at each visit with frequency calculated per 2-week periods. Cross-sectional plasma peak levels of CBD were obtained ~4 h after dosing in consecutively presenting patients. We evaluated 56 adults and 44 children (100 total; 54 female) at two time points - one before initiating CBD and one at the time of CBD plasma level testing. There was a positive linear correlation between CBD dosage (range from 5 to 50 mg/kg/day) and level (range from 7.1-1200 ng/mL) in all participants (r = 0.640; p < 0.001). The quantile regression model supported the notion of increased CBD levels being associated with improvement in seizure frequency after adjusting for age - specifically, a 100 ng/mL increase in CBD level was associated with approximately two counts reduction in seizure frequency per time period (1.87 96% confidence interval [CI] 0.34-3.39; p = 0.018). In participants with the same CBD level, differences in seizure improvement did not depend on age (p = 0.318). In this open-label study, we found evidence of a linear correlation between CBD dosage and plasma levels, and that higher dose/levels are associated with a higher response rate for seizure improvement. Children and adults responded to CBD similarly. However, seizure control response rates suggest children may respond to lower dosages/plasma levels than adults. Findings reported in this study are specific to Epidiolex® and should not be extrapolated to other CBD products.


Oxidative phosphorylation supercomplexes and respirasome reconstitution of the colorless alga Polytomella sp.
The proposal that the respiratory complexes can associate with each other in larger structures named supercomplexes (SC) is generally accepted. In the last decades most of the data about this association came from studies in yeasts, mammals and plants, and information is scarce in other lineages. Here we studied the supramolecular association of the F1FO-ATP synthase (complex V) and the respiratory complexes I, III and IV of the colorless alga Polytomella sp. with an approach that involves solubilization using mild detergents, n-dodecyl-β-D-maltoside (DDM) or digitonin, followed by separation of native protein complexes by electrophoresis (BN-PAGE), after which we identified oligomeric forms of complex V (mainly V2 and V4) and different respiratory supercomplexes (I/IV6, I/III4, I/IV). In addition, purification/reconstitution of the supercomplexes by anion exchange chromatography was also performed. The data show that these complexes have the ability to strongly associate with each other and form DDM-stable macromolecular structures. The stable V4 ATPase oligomer was observed by electron-microscopy and the association of the respiratory complexes in the so-called "respirasome" was able to perform in-vitro oxygen consumption.


Community pharmacist led medication reviews in the UK: A scoping review of the medicines use review and the new medicine service literatures.
Medicines Use Reviews (MURs) and the New Medicine Service (NMS) are services delivered by UK community pharmacists to improve adherence, improve patient understanding of their medicines and reduce medicines wastage. In this scoping review we aim to identify, map and critically examine the nature of existing empirical evidence in peer reviewed journals relating to MUR and NMS consultations. Systematic searches identified the available MUR and NMS empirical literature. We sought data on barriers and facilitators to conducting MUR or NMS consultations, the perceptions of pharmacists and patients, the conduct of consultations, and outcomes of consultations. Searches from 2005 (when MURs were introduced) to May 2018 were conducted in MEDLINE, PsycINFO, Embase and Scopus databases. Data were extracted into Excel for examination of study characteristics, participant characteristics, type of intervention/services delivered and key study quantitative and/or qualitative findings. Forty-one papers from 37 studies met the inclusion criteria: 28 papers were of MURs, 10 of NMS and 3 for both services. Studies focused on the introduction and implementation of these services, with little attention to outcomes for patients; effectiveness was not evaluated beyond in a single NMS RCT. Observational data indicated that pharmacists and patients view MURs and the NMS positively, despite challenges implementing these services and apparent lack of communication between pharmacists and GPs. Consultations were reported to be short, typically 10-12 min, characterised by limited engagement with patients and their health problems. The extent and nature of advice on health behaviours during consultations or other content was rarely examined. The research literature on MURs and the NMS has developed slowly. There is much scope for further research attention to developing more patient-centred care.


Expression of microRNAs in diffuse large B cell lymphoma is associated with immunophenotype, survival and transformation from follicular lymphoma.
MicroRNAs are naturally occurring small RNA species that regulate gene expression and are frequently abnormally expressed in cancers. However, the role of microRNAs in lymphoma is poorly understood. Therefore, we undertook a comprehensive study of microRNA expression in two of the most common lymphomas: diffuse large B-cell lymphoma (DLBCL) (n = 80) and follicular lymphoma (FCL) (n = 18) using microarrays containing probes for 464 human microRNAs. Unsupervised cluster analysis revealed distinct expression patterns between these two lymphomas and specific microRNA signatures (including members of the miR-17-92 cluster) were derived that correctly predicted lymphoma type in >95% of cases. Furthermore, we identified microRNAs in de novo DLBCL (n = 64) associated with germinal centre-like and non-germinal centre-like immunophenotypes, international prognostic index status and event-free survival in CHOP and rituximab (R)-CHOP treated patients. Despite the indolent nature of FCL a significant proportion of cases undergo high-grade transformation to more aggressive DLBCL. In order to see if transformation is associated with changes in microRNA expression we compared transformed DLBCL cases (n = 16) with de novo DLBCL, as well as FCL cases that underwent subsequent transformation (n = 7) with FCL cases that had not transformed at a median follow-up of 60 months (n = 11). Differential expression of 12 microRNAs correctly predicted >85% of transformed versus de novo DLBCL cases; six microRNAs (miR-223, 217, 222, 221 and let-7i and 7b) were found which could similarly predict or transformation in FCL (P < 0.05). These data suggest that microRNAs have potential as diagnostic and prognostic markers in these lymphomas and may be used to identify FCL patients at risk of high-grade transformation.


A Small Motor Cortex Lesion Abolished Ocular Dominance Plasticity in the Adult Mouse Primary Visual Cortex and Impaired Experience-Dependent Visual Improvements.
It was previously shown that a small lesion in the primary somatosensory cortex (S1) prevented both cortical plasticity and sensory learning in the adult mouse visual system: While 3-month-old control mice continued to show ocular dominance (OD) plasticity in their primary visual cortex (V1) after monocular deprivation (MD), age-matched mice with a small photothrombotically induced (PT) stroke lesion in S1, positioned at least 1 mm anterior to the anterior border of V1, no longer expressed OD-plasticity. In addition, in the S1-lesioned mice, neither the experience-dependent increase of the spatial frequency threshold ("visual acuity") nor of the contrast threshold ("contrast sensitivity") of the optomotor reflex through the open eye was present. To assess whether these plasticity impairments can also occur if a lesion is placed more distant from V1, we tested the effect of a PT-lesion in the secondary motor cortex (M2). We observed that mice with a small M2-lesion restricted to the superficial cortical layers no longer expressed an OD-shift towards the open eye after 7 days of MD in V1 of the lesioned hemisphere. Consistent with previous findings about the consequences of an S1-lesion, OD-plasticity in V1 of the nonlesioned hemisphere of the M2-lesioned mice was still present. In addition, the experience-dependent improvements of both visual acuity and contrast sensitivity of the open eye were severely reduced. In contrast, sham-lesioned mice displayed both an OD-shift and improvements of visual capabilities of their open eye. To summarize, our data indicate that even a very small lesion restricted to the superficial cortical layers and more than 3mm anterior to the anterior border of V1 compromised V1-plasticity and impaired learning-induced visual improvements in adult mice. Thus both plasticity phenomena cannot only depend on modality-specific and local nerve cell networks but are clearly influenced by long-range interactions even from distant brain regions.


Study of antiradical mechanisms with dihydroxybenzenes using reaction force and reaction electronic flux.
Phenolic compounds represent an important category of antioxidants because they help inhibit the oxidation process of organic compounds, while also acting as antiradicals in many biological processes. In this work, we analyze the transfer mechanisms for a set of catechols and resorcinols of a single electron, proton and hydrogen, with the radical peroxyl (˙OOH) and with different electron withdrawing and donating groups as substituents. By using the M05-2X exchange correlation functional within the Density Functional Theory framework combined with the 6-311++G(d,p) basis set, we were able to compute the Gibbs free energies for all mechanisms and compounds. According to the thermodynamic results, the hydrogen atom transfer mechanism was the most favorable. Therefore, this mechanism with substituents -CH3 and -COH in catechol and resorcinol was analyzed, using the reaction force and reaction electronic flux to characterize the structural and electronic changes that take place during the reaction. Our results show that electron donating groups favor electronic changes along the reaction path, increasing the spontaneity of the hydrogen atom transfer mechanism.


Striking a Balance: Reader Takeaways and Preferences when Integrating Text and Charts.
While visualizations are an effective way to represent insights about information, they rarely stand alone. When designing a visualization, text is often added to provide additional context and guidance for the reader. However, there is little experimental evidence to guide designers as to what is the right amount of text to show within a chart, what its qualitative properties should be, and where it should be placed. Prior work also shows variation in personal preferences for charts versus textual representations. In this paper, we explore several research questions about the relative value of textual components of visualizations. 302 participants ranked univariate line charts containing varying amounts of text, ranging from no text (except for the axes) to a written paragraph with no visuals. Participants also described what information they could take away from line charts containing text with varying semantic content. We find that heavily annotated charts were not penalized. In fact, participants preferred the charts with the largest number of textual annotations over charts with fewer annotations or text alone. We also find effects of semantic content. For instance, the text that describes statistical or relational components of a chart leads to more takeaways referring to statistics or relational comparisons than text describing elemental or encoded components. Finally, we find different effects for the semantic levels based on the placement of the text on the chart; some kinds of information are best placed in the title, while others should be placed closer to the data. We compile these results into four chart design guidelines and discuss future implications for the combination of text and charts.


The age-dependent decline of attention deficit hyperactivity disorder: a meta-analysis of follow-up studies.
This study examined the persistence of attention deficit hyperactivity disorder (ADHD) into adulthood. We analyzed data from published follow-up studies of ADHD. To be included in the analysis, these additional studies had to meet the following criteria: the study included a control group and it was clear from the methods if the diagnosis of ADHD included subjects who did not meet full criteria but showed residual and impairing signs of the disorder. We used a meta-analysis regression model to separately assess the syndromatic and symptomatic persistence of ADHD. When we define only those meeting full criteria for ADHD as having 'persistent ADHD', the rate of persistence is low, approximately 15% at age 25 years. But when we include cases consistent with DSM-IV's definition of ADHD in partial remission, the rate of persistence is much higher, approximately 65%. Our results show that estimates of ADHD's persistence rely heavily on how one defines persistence. Yet, regardless of definition, our analyses show that evidence for ADHD lessens with age. More work is needed to determine if this reflects true remission of ADHD symptoms or is due to the developmental insensitivity of diagnostic criteria for the disorder.


Pain relief induced by repetitive transcranial magnetic stimulation of precentral cortex.
Chronic electrical stimulation of the precentral (motor) cortex using surgically implanted electrodes is performed to treat medication-resistant neurogenic pain. The goal of this placebo-controlled study was to obtain such antalgic effects by means of a non-invasive cortical stimulation using repetitive transcranial magnetic stimulation (rTMS). Eighteen patients with intractable neurogenic pain of various origins were included and underwent a 20 min session of either 10 Hz, 0.5 Hz or* sham rTMS over the motor cortex in a random order. A significant decrease in the mean pain level of the series was obtained only after 10 Hz rTMS. This study shows that a transient pain relief can be induced by 10 Hz rTMS of the motor cortex in some patients suffering from chronic neurogenic pain.


Sweat: a potential marker of clinical activity in panic disorder.
Panic disorder (PD) is a paroxysmal neuropsychiatric disorder with unclear etiology and obscure pathophysiology. Despite the frequency of its occurrence, PD still has no reliable laboratory markers. The sweat is a neglected human secrete reacting immediately to various neurovegetative challenges including psychic imupulses. We hypothesized a possible dysfunction of sweat homeosthasis in PD. 10 patients with active PD, 9 patients with PD in remission and 11 age-matched controls participated in this study. All subjects underwent a single 8-min session in the dry-heat sauna. Sweat and venous blood have been collected immediately after the end of this session. Concentrations of lactate, glucose, creatinine, natrium, potassium, chlorine, calcium and magnesium have been quantitatively estimated in both liquids and compared statistically among three groups. We did not find any significant difference in blood parameters of the three above groups. However, the patients with active PD had significantly higher sweat levels of lactate, glucose, creatinine and magnesium than both the other groups which did not differ. Moreover, sweat concentrations of natrium, potassium and chlorine were significantly higher in active PD comparing to the group of PD patients in remission. The sweat of patients with active PD in comparison to PD in its clinical remission exhibits surprisingly distinctive changes of selected parameters after dry-heat sauna exposure. Increased concentrations of lactate, glucose and magnesium in the sweat are not contradictory with presupposed neurotransmitter-metabolic firing mechanisms in PD. These findings appear to be perspective biochemical markers in PD and its course.


The relationship between suboptimal effort and post-concussion symptoms in children and adolescents with mild traumatic brain injury.
This retrospective chart review study explored the relationship between suboptimal effort and post-concussion symptoms in pediatric mild traumatic brain injury (mTBI). Participants were 382 clinically referred children and adolescents between 8 and 16 years of age who sustained an mTBI. Suboptimal effort was identified using reliable digit span and age-corrected scaled scores from the Numbers subtest of the Children's Memory Scale (CMS); 20% of the sample were classified as non-credible performers. Chi-square analyses and t-tests were used to examine differences in post-concussion symptoms and neuropsychological test performance between credible and non-credible performers. Linear regression was used to examine whether CMS Numbers performance predicted post-concussion symptoms after controlling for baseline symptoms and other relevant demographic- and injury-related factors. We found that non-credible performers presented with a greater number of post-concussion symptoms as compared with credible performers. Additionally, non-credible performers demonstrated comparatively poorer performance on neuropsychological tests of focused attention and processing speed. These results suggest that children and adolescents with mTBI who fail effort testing might have a greater tendency to exaggerate post-concussion symptoms and cognitive impairment. The clinical implications of these findings are discussed.


The Ability of Secretory Leukocyte Protease Inhibitor to Inhibit Apoptosis in Monocytes Is Independent of Its Antiprotease Activity.
Secretory Leukocyte Protease Inhibitor (SLPI) is a serine protease inhibitor produced by epithelial and myeloid cells with anti-inflammatory properties. Research has shown that SLPI exerts its anti-inflammatory activity by directly binding to NF-κB DNA binding sites and, in so doing, prevents binding and subsequent transcription of proinflammatory gene expression. In the current study, we demonstrate that SLPI can inhibit TNF-α-induced apoptosis in U937 cells and peripheral blood monocytes. Specifically, SLPI inhibits TNF-α-induced caspase-3 activation and DNA degradation associated with apoptosis. We go on to show that this ability of SLPI to inhibit apoptosis is not dependent on its antiprotease activity as antiprotease deficient variants of SLPI can also inhibit TNF-α-induced apoptosis. This reduction in monocyte apoptosis may preserve monocyte function during inflammation resolution and promote infection clearance at mucosal sites.


Major depression disorder trajectories and HIV disease progression: results from a 6-year outpatient clinic cohort.
Contradictory evidence exists on the role of Major depression disorder (MDD) as a predictor of human immunodeficiency virus (HIV) disease progression, particularly regarding the effect of MDD presence versus pattern of illness. The objective of this study was to examine whether MDD status and pattern of illness differentially predict HIV disease progression. Retrospective cohort data from a six-year follow-up of HIV patients at an outpatient clinic were analyzed. MDD trajectories were identified by latent class growth analysis and generalized linear mixed models were used to examine their relation to low CD4+ T-lymphocyte counts (<200 cells/μL) during follow-up. Among 1,494 HIV patients, four MDD trajectory groups were identified: Low-Chronic, Moderate-Ascending, High-Episodic, and High-Chronic. Trajectory group membership was predicted by male sex (P = .04), minority race (P < .01), older age (P < .01) and low baseline CD4 count (P = .04). The High-Chronic group had lower odds of having a low CD4 count than the Low-Chronic group (adjusted Odds Ratio [aOR]: 0.63; 95%CI: 0.49-0.81) while the Moderate-Ascending group had higher odds (aOR: 1.53; 95%CI: 1.08-2.19). The odds of having a low CD4 count were higher among male (aOR: 1.25; 95%CI: 1.03-1.52), minority races (American Indian [aOR: 1.85; 95%CI: 1.38-2.49] and African Americans [aOR: 1.58; 95%CI: 1.33-1.87]), Hispanic (aOR: 1.52; 95%CI: 1.06-2.18), and divorced/separated patients (aOR: 1.62; 95%CI: 1.16-2.28) but decreased over time (P < .01) across trajectory groups. In this study, because MDD trajectories and CD4 counts were determined based on secondary data abstracted from electronic medical records, the results should be interpreted cautiously due to the potential for selection and misclassification bias. Overall, study findings suggest the pattern of MDD illness among HIV patients can be classified into clinically meaningful trajectory groups that appear to be programmed by known risk factors, and are useful for predicting HIV disease progression. Targeted interventions among at-risk patients may be critical to altering MDD illness patterns and curtailing HIV disease progression.


The impact of attrition in an 11-year prospective longitudinal study of younger women.
To investigate the impact of attrition on prevalence and associations between variables across four waves of a longitudinal study. Prevalence of socio-demographic and health characteristics were estimated for respondents to one, two, three or all four waves of the Australian Longitudinal Study of Women's Health cohort born between 1973 and 1978. Associations with self-rated General Health (GH) and Mental Health (MH) were compared using fixed effects in separate mixed models for respondents to at least one wave, at least two waves, at least three waves, or four waves of the longitudinal study. 14,247 women aged 18-23 years responded to Wave 1 in 1996. Respondents to all waves were more educated, and less likely to be stressed about money, to be smokers and to have children than respondents to some waves. Across all models, better GH was consistently associated with more education, no monetary stress, being married, having children, fewer visits to the doctor and not smoking. Similar results were obtained for MH. Although the potential for bias due to attrition must be considered, the current paper contributes to the growing body of evidence that suggests such biases are insufficient to preclude meaningful longitudinal analyses.


The influence of glacial meltwater on alpine aquatic ecosystems: a review.
The recent and rapid recession of alpine glaciers over the last 150 years has major implications for associated aquatic communities. Glacial meltwater shapes many of the physical features of high altitude lakes and streams, producing turbid environments with distinctive hydrology patterns relative to nival systems. Over the past decade, numerous studies have investigated the chemical and biological effects of glacial meltwater on freshwater ecosystems. Here, we review these studies across both lake and stream ecosystems. Focusing on alpine regions mainly in the Northern Hemisphere, we present examples of how glacial meltwater can affect habitat by altering physical and chemical features of aquatic ecosystems, and review the subsequent effects on the biological structure and function of lakes and streams. Collectively or separately, these factors can drive the overall distribution, diversity and behavior of primary producers, triggering cascading effects throughout the food web. We conclude by proposing areas for future research, particularly in regions where glaciers are soon projected to disappear.


Mastectomy or lumpectomy? Helping women make informed choices.
To develop an instrument to help clinicians inform their patients about surgical treatment options for the treatment of breast cancer and to evaluate the impact of the instrument on the clinical encounter. We developed an instrument, called the Decision Board, to present information regarding the benefits and risks of breast-conserving therapy (lumpectomy plus radiation therapy) and mastectomy to women with early-stage breast cancer to enable them to express a preference for the type of surgery. Seven surgeons from different communities in Ontario administered the instrument to women with newly diagnosed clinical stage I or II breast cancer over an 18-month period. Patients and surgeons were interviewed regarding acceptability of the instrument. The rates of breast-conserving surgery performed by surgeons before and after the introduction of the instrument were compared. The Decision Board was administered to 175 patients; 98% reported that the Decision Board was easy to understand, and 81% indicated that it helped them make a decision. The average score on a true/false test of comprehension was 11.8 of 14 (84%) (range, 6 to 14). Surgeons found the Decision Board to be helpful in presenting information to patients in 91% of consultations. The rate of breast-conserving surgery decreased when the Decision Board was introduced (88% v 73%, P =.001) The Decision Board is a simple method to improve communication and facilitate shared decision making. It was well accepted by patients and surgeons and easily applied in the community.


Xv4-vrxv4: a new gene-for-gene interaction identified between Xanthomonas campestris pv. vesicatoria race T3 and wild tomato relative Lycopersicon pennellii.
Strains of tomato race 3 (T3) of Xanthomonas campestris pv. vesicatoria elicit a hypersensitive response (HR) in leaves of Lycopersicon pennellii LA716. Genetic segregation of the resistance exhibited ratios near 3:1 in F2 populations, which confirmed that a single dominant gene controlled the inheritance of this trait. With the aid of a collection of introgression lines, restriction fragment length polymorphism, and cleaved amplified polymorphic sequence markers, the resistance locus was located on chromosome 3 between TG599 and TG134. An avirulence gene named avrXv4 was also isolated by mobilizing a total of 600 clones from a genomic DNA library of the T3 strain 91-118 into the X. campestris pv. vesicatoria strain ME90, virulent on L. pennellii. One cosmid clone, pXcvT3-60 (29-kb insert), induced HR in resistant plants. The avirulent phenotype of pXcvT3-60 was confirmed by comparing growth rates in planta and electrolyte leakages among transconjugants carrying a mutated or intact clone with the wild-type T3 strain 91-118. A 1.9-kb DNA fragment contained within a 6.8-kb active subclone was sequenced and was determined to carry an open reading frame of 1,077 bp. The predicted AvrXv4 protein exhibits high similarity to members of an emerging new family of bacterial proteins from plant and mammalian pathogens comprising AvrRxv, AvrBsT, YopJ, YopP, AvrA, and YL40.


Imaging characteristics, tissue distribution, and spread of a novel oncolytic vaccinia virus carrying the human sodium iodide symporter.
Oncolytic viruses show promise for treating cancer. However, to assess therapy and potential toxicity, a noninvasive imaging modality is needed. This study aims to determine the in vivo biodistribution, and imaging and timing characteristics of a vaccinia virus, GLV-1h153, encoding the human sodium iodide symporter (hNIS. GLV-1h153 was modified from GLV-1h68 to encode the hNIS gene. Timing of cellular uptake of radioiodide (131)I in human pancreatic carcinoma cells PANC-1 was assessed using radiouptake assays. Viral biodistribution was determined in nude mice bearing PANC-1 xenografts, and infection in tumors confirmed histologically and optically via Green Fluorescent Protein (GFP) and bioluminescence. Timing characteristics of enhanced radiouptake in xenografts were assessed via (124)I-positron emission tomography (PET). Detection of systemic administration of virus was investigated with both (124)I-PET and 99m-technecium gamma-scintigraphy. GLV-1h153 successfully facilitated time-dependent intracellular uptake of (131)I in PANC-1 cells with a maximum uptake at 24 hours postinfection (P<0.05). In vivo, biodistribution profiles revealed persistence of virus in tumors 5 weeks postinjection at 10(9) plaque-forming unit (PFU)/gm tissue, with the virus mainly cleared from all other major organs. Tumor infection by GLV-1h153 was confirmed via optical imaging and histology. GLV-1h153 facilitated imaging virus replication in tumors via PET even at 8 hours post radiotracer injection, with a mean %ID/gm of 3.82 ± 0.46 (P<0.05) 2 days after intratumoral administration of virus, confirmed via tissue radiouptake assays. One week post systemic administration, GLV-1h153-infected tumors were detected via (124)I-PET and 99m-technecium-scintigraphy. GLV-1h153 is a promising oncolytic agent against pancreatic cancer with a promising biosafety profile. GLV-1h153 facilitated time-dependent hNIS-specific radiouptake in pancreatic cancer cells, facilitating detection by PET with both intratumoral and systemic administration. Therefore, GLV-1h153 is a promising candidate for the noninvasive imaging of virotherapy and warrants further study into longterm monitoring of virotherapy and potential radiocombination therapies with this treatment and imaging modality.


Elevated levels of FOXA1 facilitate androgen receptor chromatin binding resulting in a CRPC-like phenotype.
Castration-resistant prostate cancer (CRPC) continues to pose a significant clinical challenge with new generation second-line hormonal therapies affording limited improvement in disease outcome. As the androgen receptor (AR) remains a critical driver in CRPC, understanding the determinants of its transcriptional activity is important for developing new AR-targeted therapies. FOXA1 is a key component of the AR transcriptional complex yet its role in prostate cancer progression and the relationship between AR and FOXA1 are not completely resolved. It is well established that FOXA1 levels are elevated in advanced prostate cancer and metastases. We mimicked these conditions by overexpressing FOXA1 in the androgen-responsive LNCaP prostate cancer cell line and observed a significant increase in AR genomic binding at novel regions that possess increased chromatin accessibility. High levels of FOXA1 resulted in increased proliferation at both sub-optimal and high 5α-dihydrotestosterone (DHT) concentrations. Immunohistochemical staining for FOXA1 in a clinical prostate cancer cohort revealed that high FOXA1 expression is associated with shorter time to biochemical recurrence after radical prostatectomy (hazard ratio (HR) 5.0, 95% confidence interval (CI) 1.2-21.1, P=0.028), positive surgical margins and higher stage disease at diagnosis. The gene expression program that results from FOXA1 overexpression is enriched for PTEN, Wnt and other pathways typically represented in CRPC gene signatures. Together, these results suggest that in an androgen-depleted state, elevated levels of FOXA1 enhance AR binding at genomic regions not normally occupied by AR, which in turn facilitates prostate cancer cell growth.


The NDR/LATS kinase Cbk1 controls the activity of the transcriptional regulator Bcr1 during biofilm formation in Candida albicans.
In nature, many microorganisms form specialized complex, multicellular, surface-attached communities called biofilms. These communities play critical roles in microbial pathogenesis. The fungal pathogen Candida albicans is associated with catheter-based infections due to its ability to establish biofilms. The transcription factor Bcr1 is a master regulator of C. albicans biofilm development, although the full extent of its regulation remains unknown. Here, we report that Bcr1 is a phosphoprotein that physically interacts with the NDR kinase Cbk1 and undergoes Cbk1-dependent phosphorylation. Mutating the two putative Cbk1 phosphoacceptor residues in Bcr1 to alanine markedly impaired Bcr1 function during biofilm formation and virulence in a mouse model of disseminated candidiasis. Cells lacking Cbk1, or any of its upstream activators, also had reduced biofilm development. Notably, mutating the two putative Cbk1 phosphoacceptor residues in Bcr1 to glutamate in cbk1Δ cells upregulated the transcription of Bcr1-dependent genes and partially rescued the biofilm defects of a cbk1Δ strain. Therefore, our data uncovered a novel role of the NDR/LATS kinase Cbk1 in the regulation of biofilm development through the control of Bcr1.


Results of reoperation for persistent and recurrent hyperparathyroidism.
Between August 1975 and January 1981, 106 patients thought to have persistent or recurrent hyperparathyroidism underwent a total of 108 parathyroid re-explorations at the National Institutes of Health. These 106 patients had a total of 175 previous operations for hyperparathyroidism (156 cervical and 19 mediastinal). Nephrolithiasis (54% of patients) and bone disease (24% o patients) were the predominant symptoms. Arteriographic examination and selective venous sampling provided highly accurate localizing results in 33% of the patients, and were of some help in 64%. The final diagnoses after reoperation and re-evaluation were: single-gland disease in 58 patients, primary nonfamilial hyperplasia in 19 patients, familial hyperplasia in three patients, multiple endocrine neoplasia (MEN) Type I in ten patients, MEN Type II in two, parathyroid carcinoma in four patients, secondary hyperplasia in three patients, and familial hypocalciuric hypercalcemia (FHH) in two patients. The diagnosis was in doubt in five patients. In the 95 patients with unequivocal hyperparathyroidism, not due to parathyroid carcinoma, surgery eliminated hypercalcemia in 91 (96%). Two patients died after operation, one of disseminated candidiasis, and one patient, with an immunodeficiency, of sepsis. Five patients developed temporary, and one permanent, recurrent nerve damage; 41% of the patients were hypocalcemic, at the time of discharge from the hospital.


Pathways linking childhood maltreatment and adult physical health.
This study examined whether a self-reported history of childhood maltreatment (physical, emotional, and sexual abuse and physical and emotional neglect) is related to poor adult physical health through health risk behaviors (obesity, substance dependence, and smoking), adverse life events, and psychological distress. Two hundred and seventy nine (279) women aged 31-54, primarily poor, urban, and African American with a history of substance use during pregnancy, were assessed for perceived physical health status using the Health Status Questionnaire (SF-36) and any reported chronic medical condition. Hierarchical multiple and logistic regression were used to test mediation, as well as to assess relative contributions of multiple mediators on physical health. More than two-thirds (n = 195, 70%) of the sample reported at least 1 form of childhood maltreatment, with 42% (n = 110) having a lifetime history of substance dependence and 59% (n = 162) having a chronic medical condition. Controlling for age, education, and race, childhood maltreatment was related to increased likelihood of lifetime history of substance dependence (OR = 1.19, 95% CI = 1.01-1.39), more adverse life events (β = .14), and greater psychological distress (β = .21). Psychological distress and adverse life events partially mediated the relationship between childhood maltreatment and perceived physical health, accounting for 42% of the association between childhood maltreatment and perceived physical health. Adverse life events accounted for 25% of the association between childhood maltreatment and chronic medical condition. Our findings provide additional evidence that the ill health effects associated with childhood maltreatment persist into adulthood. Adverse life events and psychological distress were key mechanisms shaping later physical health consequences associated with childhood maltreatment among relatively young urban women with a history of substance use. Health care providers should be aware that childhood maltreatment contributes to adult health problems. Interventions aimed at preventing child maltreatment and addressing life stress and psychological distress will improve long-term physical health among abused children, adults with such histories, as well as children who are likely to be affected by maternal history of childhood maltreatment.


Hepatitis C virus infection in chronic liver disease in Somalia.
To assess the role of hepatitis C virus (HCV) in liver disease in Somalia, antibody to HCV (anti-HCV) was studied by enzyme-linked immunosorbent assay (ELISA) and recombinant immunoblot assay (RIBA) in 110 patients with chronic liver diseases, in 309 healthy adults, in 179 institutionalized subjects with a high prevalence of intestinal parasites and Schistosoma haematobium, and in 287 children with diseases other than hepatitis. According to the RIBA test, anti-HCV was present in three healthy adults (0.97%), in four institutionalized individuals (2.2%), but in none of the children. The prevalence of anti-HCV was 4.8% in patients with hepatitis B surface antigen (HBsAg)-positive chronic liver diseases and 20.6% in patients with HBsAg-negative chronic liver diseases. Thus, HCV infection appears to play a minor role in HBsAg-positive liver disease in Somalia but may be an important factor in HBsAg-negative chronic liver disease. The low anti-HCV prevalence in individuals with no hepatic disorders is consistent with the fact that HCV does not spread by nonpercutaneous transfer. We found also a large proportion of both patients with hepatic disease and institutionalized individuals who tested positive by ELISA but not confirmed by RIBA. However, the likelihood of a true positive result increases proportionally with the ELISA value; thus, in most cases a low ELISA value probably represents a false-positive reaction, while a high ELISA value probably represents a true positive reaction.


Clearance of antitransferrin receptor immunotoxin from the rabbit eye.
Antitransferrin receptor immunotoxin has potent antiproliferative effects on proliferating human ocular cells in vitro, and shows promise in the treatment of ocular proliferative diseases, such as proliferative vitreoretinopathy, bleb scarring after trabeculectomy, or corneal epithelial downgrowth syndrome. Before treating patients, the pharmacokinetics of immunotoxin must be known. The purpose of this study was to determine the clearance of antitransferrin receptor immunotoxin from the rabbit eye after ocular injection. Immunotoxin (1,000 ng/0.1 ml PBS) was injected into the vitreous cavity of gas vitrectomized rabbit eyes. Immunotoxin in aqueous, vitreous, and plasma was measured by ELISA at various times after injection. Similar measurements were obtained after immunotoxin was injected into the anterior chamber or subconjunctival space of eyes which had not received vitrectomy. Cultured human retinal pigment epithelial cells were exposed to vitreous fluid obtained after intravitreal injection, and counted after four days, to determine the bioactivity of immunotoxin recovered from vitreous samples. Immunotoxin was cleared rapidly from the vitreous cavity over the first 24 hr (t1/2 = 8.0 hr), and thereafter was slow. Drug recovered from the vitreous cavity retained significant antiproliferative activity at 96 hr. Immunotoxin was cleared within 6 hr from the aqueous after anterior chamber injection, and was not present inside the eye after subconjunctival injection. The plasma did not contain immunotoxin after any of these injections. The favorable pharmacokinetics of antitransferrin receptor immunotoxin in the rabbit eye suggests that it could be useful in the treatment of ocular proliferative disorders.


Methylation-enhanced binding of Sp1 to the stage selector element of the human gamma-globin gene promoter may regulate development specificity of expression.
The human gamma-globin gene promoter contains a stage selector element (SSE) responsible for preferential interaction of the promoter with a powerful erythroid-specific enhancer in the fetal developmental stage (S.M. Jane, P.A. Ney, E.F. Vanin, D.L. Gumucio, and A.W. Nienhuis. EMBO J. 11:2691-2699, 1992). The element binds two proteins, the ubiquitous activator Sp1 and a protein previously known as -50 gamma and now named the stage selector protein (SSP). Binding of the second protein correlates with SSE activity in transient-transfection assays. We now report that a de novo binding site for the SSP is created by the -202(C-->G) mutation that causes hereditary persistence of fetal hemoglobin (HPFH). This site functions in an analogous manner to the SSE in hybrid beta-promoter/reporter gene constructs transfected into K562 cells. In contrast, the wild-type -202 sequence, which fails to bind the SSP, is incapable of activating the beta-gene promoter. Both the -50 and -202 HPFH sites for SSP binding overlap a consensus sequence for the transcriptional regulator Sp1. In addition, both sites contain CpG dinucleotides that are contact bases for SSP. Since the gamma promoter is known to be hypomethylated in fetal cells but fully methylated at CpG residues in adult erythroid cells, we examined the effects of this DNA modification on protein binding to the two regions. Gel mobility shift assays with nuclear extract from K562 cells (which contain both Sp1 and SSP) demonstrate preferential binding of SSP to the SSE and HPFH sites under conditions in which probe was limiting. Methylation of the CpG residues reverses this preference only in the SSE site, with a marked increase in the binding of Sp1 at the expense of the SSP. Purified Sp1 binds with 10-fold higher affinity to the methylated than to the nonmethylated -50 probe but with the same affinity to the -202 HPFH probe. The methylation-induced preferential binding of Sp1 to the SSE at the expense of SSP may be part of the mechanism by which the gamma genes are repressed in normal adult erythroid cells. In cells containing the -202 HPFH mutation, the inability of Sp1 to displace SSP in the methylated state may explain the persistence of gamma-promoter activity and gamma-gene expression observed in adults with this mutation.


Clinical and Economic Benefits of Upper Airway Stimulation for Obstructive Sleep Apnea in a European Setting.
Upper airway stimulation (UAS) is a treatment approach for patients with moderate-to-severe obstructive sleep apnea who cannot adhere to continuous positive airway pressure therapy. The objective was to evaluate added patient benefit and cost-effectiveness of UAS in the German health care system. We used a decision-analytic Markov model to project major adverse cardiovascular or cerebrovascular events (myocardial infarction [MI] or stroke), motor vehicle collision (MVC), mortality, quality-adjusted life years (QALYs), and costs. The assumed reduction in the apnea-hypopnea index with UAS compared to no treatment is based on German real-world data. Other input data were derived from the literature, public statistics, and multivariate regression. Cost-effectiveness was evaluated in Euros per QALY gained, both discounted at 3%. UAS was projected to reduce event risks (10-year relative risk for stroke, MI, cardiovascular death, and MVC: 0.76, 0.64, 0.65, and 0.34, respectively), and to increase survival by 1.27 years. While the UAS strategy incurred an additional 1.02 QALYs within the patient lifetime, there were also additional costs of EUR 45,196, resulting in an incremental cost-effectiveness ratio of EUR 44,446 per QALY gained. -Conclusions: In the present model-based analysis, UAS therapy provides meaningful benefit to patient-relevant endpoints and is a cost-effective therapy in the German setting.


Sympathetic innervation of lymphoreticular organs is rate limiting for prion neuroinvasion.
Transmissible spongiform encephalopathies are commonly propagated by extracerebral inoculation of the infectious agent. Indirect evidence suggests that entry into the central nervous system occurs via the peripheral nervous system. Here we have investigated the role of the sympathetic nervous system in prion neuroinvasion. Following intraperitoneal prion inoculation, chemical or immunological sympathectomy delayed or prevented scrapie. Prion titers in spinal cords were drastically reduced at early time points after inoculation. Instead, keratin 14-NGF transgenic mice, whose lymphoid organs are hyperinnervated by sympathetic nerves, showed reduction in scrapie incubation time and, unexpectedly, much higher titers of prion infectivity in spleens. We conclude that sympathetic innervation of lymphoid organs is rate limiting for prion neuroinvasion and that splenic sympathetic nerves may act as extracerebral prion reservoirs.


Factors affecting the spatial pattern of nitrate contamination in shallow groundwater.
The elevated level of nitrate in groundwater is a serious problem in Korean agricultural areas. To control and manage groundwater quality, the characterization of groundwater contamination and identification of the factors affecting the nitrate concentration of groundwater are significant. The characterization of groundwater contamination at a hydrologically complex agricultural site in Yupori, Chuncheon (Korea) was undertaken by analyzing the hydrochemical data of groundwater within a statistical framework. Multivariate statistical tools such as cluster analyses and Tobit regression were applied to investigate the spatial variation of nitrate contamination and to analyze the factors affecting the NO3-N concentration in a shallow groundwater system. The groundwater groups from the cluster analysis were consistent with the land use pattern of the study area. The clustered group of a gentle-slope area with lower elevations showed higher NO3-N contamination of groundwater than groups on a hillside with higher elevations. Tobit regression results indicated that the agricultural activity in the vegetable fields and barns were the major factors affecting the elevated NO3-N concentration while the land slopes and elevations were negatively correlated with the NO3-N concentration. This shows that topographic characteristics such as land slopes and elevations should be considered to evaluate the land use impact on shallow groundwater quality.


A single intraoperative sub-tenon's capsule injection of triamcinolone and ciprofloxacin in a controlled-release system for cataract surgery.
To compare intraoperative injection of triamcinolone and ciprofloxacin in a controlled-release system (DuoCat) with prednisolone and ciprofloxacin eye drops after cataract surgery. In this randomized, double-masked, controlled trial, a total of 135 patients undergoing cataract surgery were randomly allocated to two groups: 67 patients treated after surgery with prednisolone 1% and ciprofloxacin 3% eye drops four times daily (week 1), three times daily (week 2), twice daily (week 3), and once daily (week 4) and 0.3% ciprofloxacin drops four times daily (weeks 1 and 2), and 68 patients treated at the end of surgery with a sub-Tenon's injection of 25 mg triamcinolone and 2 mg ciprofloxacin in biodegradable microspheres. The patients were examined on postoperative days 1, 3, 7, 14, and 28. The main outcome measures were postoperative anterior chamber cell and flare, intraocular pressure (IOP), lack of anti-inflammatory response, and presence of infection. No significant differences were observed between the groups in anterior chamber cell (P > 0.14) and flare (P > 0.02) at any postoperative visits. The mean (99% confidence interval) differences in IOP between the prednisolone and triamcinolone groups on days 1, 3, 7, 14, and 28 were -0.4 mm Hg (-2.1 to 1.3), 0.0 mm Hg (-1.4 to 1.3), 0.0 mm Hg (-1.1 to 1.1), -0.2 mm Hg (-1.1 to 0.8), and -0.1 mm Hg (-1.1 to 0.9), respectively. No patient had a postoperative infection. One injection of DuoCat had a therapeutic response and ocular tolerance that were equivalent to conventional eye drops in controlling inflammation after cataract surgery. (ClinicalTrials.gov number, NCT00431028.).


Complications of infective endocarditis.
Infective endocarditis (IE) is a lethal disease if not promptly treated with antibiotics, either in association with surgery or not. The incidence of disease has not decreased over the last decades due to the change of risk conditions. Complications of IE may involve cardiac structures when the infection spreads within the heart, or extra cardiac ones when the cause is usually from embolic origin; they may also be due to medical treatment or to the septic condition itself. A variety of complications may occur in most of patients. The literature reports one complication of IE in 57%, two in 26% and three or more in about 14% of patients examined. The frequency of specific complications depends on variables as the infecting pathogen, duration of disease before therapy and type of treatment. However it is often difficult to assess the true incidence of complications because the published reviews in literature are frequently based on retrospective chart reviews and different diagnostic criteria are used. The decision over either indication or timing of surgery should be individualized and based on a multidisciplinary approach involving at least cardiologists and cardiac surgeons. Congestive heart failure (CHF) is the most important complication of IE, which has the greatest impact on prognosis. Periannular abscesses are a relatively common complication of IE (42% to 85% of cases during surgery or at autopsy respectively), associated with a higher morbidity and mortality. Systemic embolization occurs in 22% to 50% of cases; emboli may involve major arteries, mostly affecting the central nervous system, but also other organs. Splenic abscess is a rare complication of IE, due to direct seeding of spleen by an embolus or bacterial seeding of a bland infarction. Neurological complications develop in 20% to 40% of patients with IE and represent a dangerous subset of complications. Mycotic aneurysms are rare, resulting from diffusion of infection to the vessel wall. Actually the clinical profile, the best treatment (medical or surgical approach) and outcome of complicated IE are not well defined. Changing trends in aetiology of IE with emerging infections from Staphylococci, bacteria of the HACEK group and Fungi have resulted in an increased frequency of culture negative IE. Sepsis or persistent fever despite appropriate antimicrobial therapy, recurrent emboli, heart failure or new pathologic murmurs suggest haemodynamic impairment and/or infection extending beyond the valve leaflet or prosthetic valvular annulus. The course of the disease will consequently get worse with an increasing need of surgery. Patients who develop abscesses are more likely to undergo surgery than those who do not (84-91% vs 36%), and also their in-hospital mortality rate is higher (19% vs 11%). A prompt detection of complications often allows an earlier surgical treatment which represents the best way to improve the outcome. The introduction of molecular methods techniques has increased the ability to identify the causal agents of IE, mostly in cases of culture negative endocarditis. Echocardiography, mainly from transesophageal (TEE) approach, has significantly improved the evaluation of IE allowing to detect the specific signs of the disease as vegetations, abscesses, valve insufficiency, prosthetic valve dehiscence, fistulas. In our 3rd referral Hospital (Lancisi Heart Hospital, Ancona, Italy) we performed a follow-up (mean 8.26 years) of 15 patients with periannular complications associated with IE. The long term follow-up showed low mortality rate, high incidence of reintervention, improved New York Heart Association (NYHA) class in survivors and no changes of the lesions at the echocardiographic examination, suggesting that periannular complications have not significantly influenced the overall survival in our patients at the follow-up.


Expression of melatonin MT(1) and MT(2) receptors, and ROR alpha(1) receptor in transplantable murine Colon 38 cancer.
There are some data suggesting that melatonin exerts oncostatic action through membrane as well as nuclear receptors. In previous studies we demonstrated the antiproliferative and proapoptotic action of melatonin on transplantable murine Colon 38 adenocarcinoma cells. Therefore, the aim of the present study was to determine whether the membrane melatonin receptors MT(1) and MT(2) as well as the nuclear receptor RZR/ROR alpha are expressed in Colon 38 cells. Adult male B6D2F1 mice were used in this experiment. The induction of tumor was conducted by subcutaneous injection of 0.2 mL of a 33% suspension of Colon 38 cancer cells into axillary region. Expression of mRNA encoding MT(1) and MT(2) melatonin membrane receptors was studied by RT-PCR analysis, and expression of ROR alpha(1) nuclear receptor protein was studied by Western blot analysis. The expression of mRNA encoding both MT(1) and MT(2) melatonin receptors was demonstrated in Colon 38 cancer cells. Moreover, immunodetection revealed the expression of MT(1) and ROR alpha(1) proteins in these cells. Our studies on Colon 38 adenocarcinoma cells support the concept that both membrane and nuclear receptors are involved in the oncostatic action of melatonin.


A comparative study of virginity in fig wasps
In haplodiploid species, the presence of unmated (virgin) females that can produce only haploid male offspring may have several important effects on a range of phenomena such as reproductive strategies, the transmission of parasitic chromosomes and the evolution of eusociality. The strength of these effects will depend upon the prevalence of virgin females. Two theories make conflicting predictions concerning the importance of factors that should be associated with increased levels of virginity, emphasizing either the degree of local mate competition or brood size. In this paper, a model is presented which predicts that, under conditions of local mate competition, the prevalence of virginity should be negatively correlated with the average number of offspring developing in a patch. The different predictions were then tested using data from 53 species of fig wasps representing 15 genera from four continents. Across species, the estimated prevalence of virginity was significantly inversely related to brood size, but showed no correlation with sex ratio (an index of local mate competition), supporting the predictions of our model. Qualitatively similar results were found when a formal comparative analysis was carried out using a morphologically and molecularly based phylogeny.


Progress and impact of 13 years of the global programme to eliminate lymphatic filariasis on reducing the burden of filarial disease.
A Global Programme to Eliminate Lymphatic Filariasis was launched in 2000, with mass drug administration (MDA) as the core strategy of the programme. After completing 13 years of operations through 2012 and with MDA in place in 55 of 73 endemic countries, the impact of the MDA programme on microfilaraemia, hydrocele and lymphedema is in need of being assessed. During 2000-2012, the MDA programme made remarkable achievements - a total of 6.37 billion treatments were offered and an estimated 4.45 billion treatments were consumed by the population living in endemic areas. Using a model based on empirical observations of the effects of treatment on clinical manifestations, it is estimated that 96.71 million LF cases, including 79.20 million microfilaria carriers, 18.73 million hydrocele cases and a minimum of 5.49 million lymphedema cases have been prevented or cured during this period. Consequently, the global prevalence of LF is calculated to have fallen by 59%, from 3.55% to 1.47%. The fall was highest for microfilaraemia prevalence (68%), followed by 49% in hydrocele prevalence and 25% in lymphedema prevalence. It is estimated that, currently, i.e. after 13 years of the MDA programme, there are still an estimated 67.88 million LF cases that include 36.45 million microfilaria carriers, 19.43 million hydrocele cases and 16.68 million lymphedema cases. The MDA programme has resulted in significant reduction of the LF burden. Extension of MDA to all at-risk countries and to all regions within those countries where MDA has not yet reached 100% geographic coverage is imperative to further reduce the number of microfilaraemia and chronic disease cases and to reach the global target of interrupting transmission of LF by 2020.


Molecular Dynamics Simulations of Photo-Induced Free Radical Polymerization.
Curing kinetic models provide insight into how design parameters affect the kinetics of photopolymerization. However, they do not provide insight into how networks form or how they influence the process. This article describes a molecular dynamics simulation framework for simulating photoinitiated, chain growth, free radical polymerization. The framework was applied to simulate the photo-induced polymerization of bisphenol A (EO)10 diacrylate under varying conditions of curing light intensity and photoinitiator concentration. Results from the simulations agree very well with curing kinetic curves and gelation points derived from experiments. The simulations also reveal that: (1) gelation is highly correlated with the formation of giant molecules, (2) differences in the number of free radicals generated at the beginning of polymerization significantly affect polymer network formation at low to intermediate conversion, and thus affect the gelation point, and (3) increasing light intensity or photoinitiator concentration tends to delay the gelation point, but does not affect the ultimate polymer network structure near the latter stages of photopolymerization.


The Role of Qualitative Research Methods in Discrete Choice Experiments.
The use of qualitative research (QR) methods is recommended as good practice in discrete choice experiments (DCEs). This study investigated the use and reporting of QR to inform the design and/or interpretation of healthcare-related DCEs and explored the perceived usefulness of such methods. DCEs were identified from a systematic search of the MEDLINE database. Studies were classified by the quantity of QR reported (none, basic, or extensive). Authors ( n = 91) of papers reporting the use of QR were invited to complete an online survey eliciting their views about using the methods. A total of 254 healthcare DCEs were included in the review; of these, 111 (44%) did not report using any qualitative methods; 114 (45%) reported "basic" information; and 29 (11%) reported or cited "extensive" use of qualitative methods. Studies reporting the use of qualitative methods used them to select attributes and/or levels ( n = 95; 66%) and/or pilot the DCE survey ( n = 26; 18%). Popular qualitative methods included focus groups ( n = 63; 44%) and interviews ( n = 109; 76%). Forty-four studies (31%) reported the analytical approach, with content ( n = 10; 7%) and framework analysis ( n = 5; 4%) most commonly reported. The survey identified that all responding authors ( n = 50; 100%) found that qualitative methods added value to their DCE study, but many ( n = 22; 44%) reported that journals were uninterested in the reporting of QR results. Despite recommendations that QR methods be used alongside DCEs, the use of QR methods is not consistently reported. The lack of reporting risks the inference that QR methods are of little use in DCE research, contradicting practitioners' assessments. Explicit guidelines would enable more clarity and consistency in reporting, and journals should facilitate such reporting via online supplementary materials.


[Contemporary practice of the coronary angioplasty in Chile. Final report of the Chilean National Coronary Angioplasty Registry (RENAC) 2001-2002].
Coronary angioplasty was introduced in Chile in 1982, but only after the introduction of stents it became commonplace. However the results of this procedure at the national level remain largely unknown. To assess the results and characteristics of coronary angioplasty and develop a national registry in Chile (RENAC). All centers performing angioplasty were invited to contribute prospectively with the clinical, angiographic, procedural characteristics and results of all consecutive coronary angioplasty procedures attempted between June 2001 and October 2002. In 10 centers, 1768 lesions were treated in 1484 procedures (22.98% in women). Mean age was 60.8 +/- 11.3 years old. Diabetes was found in 21.8%, hypertension in 60.2%, dyslipidemia in 52.0%, active smoking in 40.0%, renal failure in 6.2%. Myocardial infarction was recent in 28.4% and acute in 19.7%. Forty eight percent of patients had unstable angina and 15.5% had stable angina. Fifty three percent of patients had single vessel disease and 71%, normal left ventricular eyection fraction. One vessel was treated in 90.6% of patients and 81.7% of lesions were treated with stents, 17.9% only with baloon and in 0.4% with atherectomy. Angiographic success was obtained in 95.2%. Clinical success was obtained in 92.2%, and 95.1% In patients without acute myocardial infarction. Overall inhospital death was 2.2%. In patients without myocardial infarction, the figure was 1.1%. Coronary angioplasty in Chile is performed mostly for the treatment of acute coronary syndromes. Stents are the most frequently used devices. The high success, low mortality and complications observed are comparable to North American registries.


Exploring the experiences of bereaved families involved in assisted suicide in Southern Switzerland: a qualitative study.
In Switzerland, helping with assisted suicide under certain conditions is not prosecuted. With approximately 300 cases annually, this leaves behind a large group of bereaved people where its consequences are mostly unknown. The study aimed to explore family involvement in decision making prior to assisted suicide, and to examine their ways of coping during the bereavement period. A qualitative interview study used the principles of Grounded Theory analysis. Eleven relatives of eight patients, who died in Southern Switzerland after assisted suicide, participated in semistructured interviews. The large majority of family members faced moral dilemmas during the decision-making phase. Their respect for patient's autonomy was a key justification to resolve dilemmas. Two types of involvement were identified: categorised as 'passive' when the decision making was located with the patient, and 'active' when assisted suicide was proposed by the family member and/or the relative was involved in some way. The relatives reported feelings of isolation during and after assisted suicide. Family members reported fear of social stigma and did not openly disclose assisted suicide as the cause of death. None of those interviewed received formal psychological support. Bereaved families express moral dilemmas, feelings of isolation and secrecy in the management of assisted suicide in Southern Switzerland. These features seem underestimated and not sufficiently recognised by the healthcare professionals. Management of assisted suicide requests should include consideration of family members' needs, in addition to those of the patient.


Gaussian predictive process models for large spatial data sets.
With scientific data available at geocoded locations, investigators are increasingly turning to spatial process models for carrying out statistical inference. Over the last decade, hierarchical models implemented through Markov chain Monte Carlo methods have become especially popular for spatial modelling, given their flexibility and power to fit models that would be infeasible with classical methods as well as their avoidance of possibly inappropriate asymptotics. However, fitting hierarchical spatial models often involves expensive matrix decompositions whose computational complexity increases in cubic order with the number of spatial locations, rendering such models infeasible for large spatial data sets. This computational burden is exacerbated in multivariate settings with several spatially dependent response variables. It is also aggravated when data are collected at frequent time points and spatiotemporal process models are used. With regard to this challenge, our contribution is to work with what we call predictive process models for spatial and spatiotemporal data. Every spatial (or spatiotemporal) process induces a predictive process model (in fact, arbitrarily many of them). The latter models project process realizations of the former to a lower dimensional subspace, thereby reducing the computational burden. Hence, we achieve the flexibility to accommodate non-stationary, non-Gaussian, possibly multivariate, possibly spatiotemporal processes in the context of large data sets. We discuss attractive theoretical properties of these predictive processes. We also provide a computational template encompassing these diverse settings. Finally, we illustrate the approach with simulated and real data sets.