Does pulmonary rehabilitation reduce anxiety and depression in chronic obstructive pulmonary disease?
Symptoms of anxiety and depression are prevalent in at least 20-40% of patients with moderate-to-severe chronic obstructive pulmonary disease (COPD) and are known to affect prognosis and worsen quality of life. This review examines whether pulmonary rehabilitation can effectively improve psychological status in COPD patients. Over the last decade, evidence has accumulated to show that comprehensive pulmonary rehabilitation, which includes exercise training and disease education or psychosocial support or both, can effectively improve anxiety and depression in COPD. The quality of supporting data varies but indicates a consistent and homogeneous effect in favour of pulmonary rehabilitation for anxiety and depression when compared with conventional community care. Pulmonary rehabilitation also appears to improve anxiety and depression in rehabilitating COPD patients with less favourable psychosocial health, but its efficacy for treating more severe and enduring anxiety and depression is largely unproven. Additionally, it is not certain which components of pulmonary rehabilitation lead to improved anxiety and depression, and further research is needed to ensure that psychological benefits accrued during rehabilitation are sustained. Anxiety and depression afflict a significant proportion of COPD patients and lead to worse patient-centred outcomes. Evidence supports the use of pulmonary rehabilitation to effectively manage psychological morbidity in COPD.


Fish intake during pregnancy and foetal neurodevelopment--a systematic review of the evidence.
Fish is a source of several nutrients that are important for healthy foetal development. Guidelines from Australia, Europe and the USA encourage fish consumption during pregnancy. The potential for contamination by heavy metals, as well as risk of listeriosis requires careful consideration of the shaping of dietary messages related to fish intake during pregnancy. This review critically evaluates literature on fish intake in pregnant women, with a focus on the association between neurodevelopmental outcomes in the offspring and maternal fish intake during pregnancy. Peer-reviewed journal articles published between January 2000 and March 2014 were included. Eligible studies included those of healthy pregnant women who had experienced full term births and those that had measured fish or seafood intake and assessed neurodevelopmental outcomes in offspring. Medline, Scopus, Web of Science, ScienceDirect and the Cochrane Library were searched using the search terms: pregnant, neurodevelopment, cognition, fish and seafood. Of 279 papers sourced, eight were included in the final review. Due to heterogeneity in methodology and measured outcomes, a qualitative comparison of study findings was conducted. This review indicates that the benefits of diets providing moderate amounts of fish during pregnancy outweigh potential detrimental effects in regards to offspring neurodevelopment. It is important that the type of fish consumed is low in mercury.


Short-form Tai Chi improves standing balance of people with chronic stroke.
Our previous findings showed that 4 weeks of intensive Tai Chi practice improved standing balance in healthy seniors. This study set out to investigate whether Tai Chi could improve standing balance in subjects with chronic stroke. One hundred thirty-six subjects >6 months after stroke were randomly assigned to a control group (n = 62) practicing general exercises or a Tai Chi group (n = 74) for 12 weeks of training. Each week, 1 hour of group practice was supplemented by 3 hours of self-practice. We used a short-form of Tai Chi consisting of 12 forms that require whole-body movements to be performed in a continuous sequence and demands concentration. A blinded assessor examined subjects at baseline, 6 weeks (mid-program), 12 weeks (end-program), and 18 weeks (follow-up). The 3 outcome measures were (1) dynamic standing balance evaluated by the center of gravity (COG) excursion during self-initiated body leaning in 4 directions, (2) standing equilibrium evaluated in sensory challenged conditions, and (3) functional mobility assessed by Timed-up-and-go score. Mixed model repeated-measures analysis of variance was used to examine between-group differences. When compared with the controls, the Tai Chi group showed greater COG excursion amplitude in leaning forward, backward, and toward the affected and nonaffected sides (P < .05), as well as faster reaction time in moving the COG toward the nonaffected side (P = .014) in the end-program and follow-up assessments. The Tai Chi group also demonstrated better reliance on vestibular integration for balance control at end-program (P = .038). However, neither group improved significantly in Timed-up-and-go scores. Twelve weeks of short-form Tai Chi produced specific standing balance improvements in people with chronic stroke that outlasted training for 6 weeks.


A biofilm model to understand the onset of sulfate reduction in denitrifying membrane biofilm reactors.
This work presents a multispecies biofilm model that describes the co-existence of nitrate- and sulfate-reducing bacteria in the H(2)-based membrane biofilm reactor (MBfR). The new model adapts the framework of a biofilm model for simultaneous nitrate and perchlorate removal by considering the unique metabolic and physiological characteristics of autotrophic sulfate-reducing bacteria that use H(2) as their electron donor. To evaluate the model, the simulated effluent H(2), UAP (substrate-utilization-associated products), and BAP (biomass-associated products) concentrations are compared to experimental results, and the simulated biomass distributions are compared to real-time quantitative polymerase chain reaction (qPCR) data in the experiments for parameter optimization. Model outputs and experimental results match for all major trends and explain when sulfate reduction does or does not occur in parallel with denitrification. The onset of sulfate reduction occurs only when the nitrate concentration at the fiber's outer surface is low enough so that the growth rate of the denitrifying bacteria is equal to that of the sulfate-reducing bacteria. An example shows how to use the model to design an MBfR that achieves satisfactory nitrate reduction, but suppresses sulfate reduction.


Detection and characterization of beta-adrenergic receptors and adenylate cyclase in coated vesicles isolated from bovine brain.
To assess whether internalization of beta-adrenergic receptor occurs in the CNS, we have isolated clathrin-coated vesicles from bovine forebrain and examined them for the presence of beta-adrenergic receptor binding and adenylate cyclase activities. A coated vesicle enriched preparation isolated by successive D2O-Ficoll density gradient centrifugations was applied to a glass bead permeation column to achieve further purification. Two major peaks of protein were eluted from the column and monitored by electron microscopy and SDS-PAGE. Peak II contained almost exclusively coated vesicles (98%), whereas peak I, which appeared in the void volume, contained larger smooth vesicles and few coated vesicles. beta-Adrenergic receptor binding to peaks I and II was measured with 125I-cyanopindolol (CYP) as ligand in Sepharose 4B column assays. 125I-CYP was found to bind specifically and saturably to both peaks I and II with a Bmax of 28 +/- 4 and 32 +/- 3 fmol/mg protein, respectively. 3H-CGP 12177, a hydrophilic beta-adrenergic receptor ligand, did not label receptors present in peak II, but it specifically bound to synaptic plasma membranes (SPM) prepared from bovine hippocampus and, to a lesser extent, to peak I. These results suggest that receptors present in coated vesicles are cryptic in nature. In the displacement of 125I-CYP binding by (-)-isoproterenol, addition of 50 microM GppNHp caused a significant "right shift" with SPM and peak I but not the peak II preparation. Adenylate cyclase activities could also be detected in both peaks I and II (specific activities, 21 +/- 0.6 and 24 +/- 0.5 pmol cAMP/mg protein/min, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Plasma phospholipid long-chain ω-3 fatty acids and total and cause-specific mortality in older adults: a cohort study.
Long-chain ω-3 polyunsaturated fatty acids (ω3-PUFAs), including eicosapentaenoic acid (EPA) (20:5ω-3), docosapentaenoic acid (DPA) (22:5ω-3), and docosahexaenoic acid (DHA) (22:6ω-3), have been shown to reduce cardiovascular risk, but effects on cause-specific and total mortality and potential dose-responses remain controversial. Most observational studies have assessed self-reported dietary intake and most randomized trials have tested effects of adding supplements to dietary intake and evaluated secondary prevention, thus limiting inference for dietary ω3-PUFAs or primary prevention. To investigate associations of plasma phospholipid EPA, DPA, DHA, and total ω3-PUFA levels with total and cause-specific mortality among healthy older adults not receiving supplements. Prospective cohort study. 4 U.S. communities. 2692 U.S. adults aged 74 years (±5 years) without prevalent coronary heart disease (CHD), stroke, or heart failure at baseline. Phospholipid fatty acid levels and cardiovascular risk factors were measured in 1992. Relationships with total and cause-specific mortality and incident fatal or nonfatal CHD and stroke through 2008 were assessed. During 30 829 person-years, 1625 deaths (including 570 cardiovascular deaths), 359 fatal and 371 nonfatal CHD events, and 130 fatal and 276 nonfatal strokes occurred. After adjustment, higher plasma levels of ω3-PUFA biomarkers were associated with lower total mortality, with extreme-quintile hazard ratios of 0.83 for EPA (95% CI, 0.71 to 0.98; P for trend = 0.005), 0.77 for DPA (CI, 0.66 to 0.90; P for trend = 0.008), 0.80 for DHA (CI, 0.67 to 0.94; P for trend = 0.006), and 0.73 for total ω3-PUFAs (CI, 0.61 to 0.86; P for trend < 0.001). Lower risk was largely attributable to fewer cardiovascular than noncardiovascular deaths. Individuals in the highest quintile of phospholipid ω3-PUFA level lived an average of 2.22 more years (CI, 0.75 to 3.13 years) after age 65 years than did those in the lowest quintile. Temporal changes in fatty acid levels and misclassification of causes of death may have resulted in underestimated associations, and unmeasured or imperfectly measured covariates may have caused residual confounding. Higher circulating individual and total ω3-PUFA levels are associated with lower total mortality, especially CHD death, in older adults. National Institutes of Health.


Psychological stress in children may alter the immune response.
Psychological stress is a public health issue even in children and has been associated with a number of immunological diseases. The aim of this study was to examine the relationship between psychological stress and immune response in healthy children, with special focus on autoimmunity. In this study, psychological stress was based on a composite measure of stress in the family across the domains: 1) serious life events, 2) parenting stress, 3) lack of social support, and 4) parental worries. PBMCs, collected from 5-y-old high-stressed children (n = 26) and from 5-y-old children without high stress within the family (n = 52), from the All Babies In Southeast Sweden cohort, were stimulated with Ags (tetanus toxoid and β-lactoglobulin) and diabetes-related autoantigens (glutamic acid decarboxylase 65, insulin, heat shock protein 60, and tyrosine phosphatase). Immune markers (cytokines and chemokines), clinical parameters (C-peptide, proinsulin, glucose), and cortisol, as an indicator of stress, were analyzed. Children from families with high psychological stress showed a low spontaneous immune activity (IL-5, IL-10, IL-13, IL-17, CCL2, CCL3, and CXCL10; p < 0.01) but an increased immune response to tetanus toxoid, β-lactoglobulin, and the autoantigens glutamic acid decarboxylase 65, heat shock protein 60, and tyrosine phosphatase (IL-5, IL-6, IL-10, IL-13, IL-17, IFN-γ, TNF-α, CCL2, CCL3, and CXCL10; p < 0.05). Children within the high-stress group showed high level of cortisol, but low level of C-peptide, compared with the control group (p < 0.05). This supports the hypothesis that psychological stress may contribute to an imbalance in the immune response but also to a pathological effect on the insulin-producing β cells.


Open for collaboration: an academic platform for drug discovery and development at SciLifeLab.
The Science for Life Laboratory Drug Discovery and Development (SciLifeLab DDD) platform reaches out to Swedish academia with an industry-standard infrastructure for academic drug discovery, supported by earmarked funds from the Swedish government. In this review, we describe the build-up and operation of the platform, and reflect on our first two years of operation, with the ambition to share learnings and best practice with academic drug discovery centers globally. We also discuss how the Swedish Teacher Exemption Law, an internationally unique aspect of the innovation system, has shaped the operation. Furthermore, we address how this investment in infrastructure and expertise can be utilized to facilitate international collaboration between academia and industry in the best interest of those ultimately benefiting the most from translational pharmaceutical research - the patients.


Prediction of protein backbone conformation based on seven structure assignments. Influence of local interactions.
A method is developed to compute backbone tertiary folds from the amino acid sequence. In this method, the number of degrees of freedom is drastically reduced by neglecting side-chain flexibility, and by describing backbone conformations as combinations of only seven structural states. These are characterized by single values of the dihedral angles phi, psi and omega, representing allowed conformations of the isolated dipeptide. We show that this restrictive model is none the less capable of describing native backbones to within acceptable deviations. Using our backbone description, potentials of mean force are derived from a database of known protein structures, based on statistical influences of single residues and residue pairs on the conformational states in their vicinity along the chain. This yields the force-field component due to local interactions, which is then used to predict lowest-energy conformations from any given amino acid sequence. The prediction algorithm does not require searching conformational space and is therefore extremely fast. Another important asset of our method is that it is able to compute not only the minimum energy conformation, but any number of lowest energy structures, whose relative preferences can be determined from the corresponding computed energy values. The performance of our procedure is tested on short peptides that are likely to be stabilized by local interactions. These include several helical structures and a hexapeptide with a beta-bend conformation, corresponding to peptides shown to have relatively well-defined conformations in aqueous solution, and to protein segments believed to adopt their native conformation early during folding. In addition, several flexible peptides are analysed. Except for the problems encountered in predicting observed disulphide bridges in two of the flexible peptides, and in a somewhat larger fragment comprising residues 30 to 51 of bovine trypsin inhibitor, prediction results compare very favourably with experimental data. Potential applications of our procedure to protein modelling and its extension to protein folding are discussed.


Modelling prokaryote gene content.
The patchy distribution of genes across the prokaryotes may be caused by multiple gene losses or lateral transfer. Probabilistic models of gene gain and loss are needed to distinguish between these possibilities. Existing models allow only single genes to be gained and lost, despite the empirical evidence for multi-gene events. We compare birth-death models (currently the only widely-used models, in which only one gene can be gained or lost at a time) to blocks models (allowing gain and loss of multiple genes within a family). We analyze two pairs of genomes: two E. coli strains, and the distantly-related Archaeoglobus fulgidus (archaea) and Bacillus subtilis (gram positive bacteria). Blocks models describe the data much better than birth-death models. Our models suggest that lateral transfers of multiple genes from the same family are rare (although transfers of single genes are probably common). For both pairs, the estimated median time that a gene will remain in the genome is not much greater than the time separating the common ancestors of the archaea and bacteria. Deep phylogenetic reconstruction from sequence data will therefore depend on choosing genes likely to remain in the genome for a long time. Phylogenies based on the blocks model are more biologically plausible than phylogenies based on the birth-death model.


Blood biomarkers MMP-7 and SP-A: predictors of outcome in idiopathic pulmonary fibrosis.
Because of the variable course of idiopathic pulmonary fibrosis (IPF), it is important to generate an accurate prognosis at the time of diagnosis. The aim of this study was to investigate the prognostic value of blood biomarkers in IPF. The plasma level of the biomarkers, matrix metalloproteinase-7 (MMP-7), Krebs von den Lungen-6 antigen, and surfactant protein (SP)-A and SP-D were retrospectively compared with the clinical course of 118 patients with IPF, 68 of whom had biopsy-proven IPF. The median follow-up period was 24 months. Multivariate Cox analysis showed MMP-7 (HR, 1.056; P = .0063) and SP-A (HR, 1.011; P = .0001) were significant predictors of survival along with age, FVC, and extent of honeycombing. The patients with high levels of both MMP-7 (≥ 12.1 ng/mL) and SP-A (≥ 80.3 ng/mL) had shorter survival (1-year survival rate: 59%) and higher frequency (42%) of lung function decline (> 10% reduction in FVC in 6 months) compared with those with high levels of one biomarker (1-year survival rate: 81%; FVC decline: 27%) or low levels of both (1-year survival rate: 83.3%; FVC decline: 9%). Multivariate models demonstrated marginal improvement in the prediction of mortality (concordance index [C-index]: 0.731; P = .061) when MMP-7 and SP-A were included and compared with standard clinical predictors only (C-index: 0.686); however, it became significant with addition of MMP-7, SP-A, and Krebs von den Lungen-6 antigen (C-index: 0.730; P = .037). Our retrospective study suggested that at least three biomarkers are necessary to improve predictability of mortality in IPF compared with clinical parameters. Further study in a greater number of patients is warranted.


A new strategy employed for identification of sweet orange cultivars with RAPD markers.
We optimized RAPD techniques by increasing the length of RAPD primers and performing a strict screening of PCR annealing temperature to distinguish 60 sweet orange cultivars from the Research Institute of Pomology at the Chinese Academy of Agricultural Sciences. A new approach called cultivar identification diagram (CID) was used to improve the efficiency of RAPD markers for cultivar identification. Thirteen effective primers were first screened from 54 RAPD arbitrary 11-mer primers based on their amplification products and amplified polymorphic bands; they were then used for PCR amplification of all 60 cultivars. All cultivars were manually and completely separated by the polymorphic bands appearing in DNA fingerprints from 13 primers; a CID of the 60 sweet orange cultivars was then constructed. This CID separated all the cultivars from each other, based on the polymorphic bands; the corresponding primers were marked in the correct positions on the sweet orange CID. The CID strategy facilitates the identification of fruit cultivars with DNA markers. This CID of sweet orange cultivars will be very useful for the protection of cultivar rights and for early identification of seedlings in the nursery industry.


Cellular senescence in osteoarthritis pathology.
Cellular senescence is a state of stable proliferation arrest of cells. The senescence pathway has many beneficial effects and is seen to be activated in damaged/stressed cells, as well as during embryonic development and wound healing. However, the persistence and accumulation of senescent cells in various tissues can also impair function and have been implicated in the pathogenesis of many age-related diseases. Osteoarthritis (OA), a severely debilitating chronic condition characterized by progressive tissue remodeling and loss of joint function, is the most prevalent disease of the synovial joints, and increasing age is the primary OA risk factor. The profile of inflammatory and catabolic mediators present during the pathogenesis of OA is strikingly similar to the secretory profile observed in 'classical' senescent cells. During OA, chondrocytes (the sole cell type present within articular cartilage) exhibit increased levels of various senescence markers, such as senescence-associated beta-galactosidase (SAβGal) activity, telomere attrition, and accumulation of p16ink4a. This suggests the hypothesis that senescence of cells within joint tissues may play a pathological role in the causation of OA. In this review, we discuss the mechanisms by which senescent cells may predispose synovial joints to the development and/or progression of OA, as well as touching upon various epigenetic alterations associated with both OA and senescence.


Evaluation of young men with organic erectile dysfunction.
Erectile dysfunction (ED) in men under the age of 40 was once thought to be entirely psychogenic. Over the last few decades, advances in our understanding of erectile physiology and improvements in diagnostic testing have restructured our understanding of ED and its etiologies. Although psychogenic ED is more prevalent in the younger population, at least 15%-20% of these men have an organic etiology. Organic ED has been shown to be a predictor of increased future morbidity and mortality. As such, a thorough work-up should be employed for any man with complaints of sexual dysfunction. Oftentimes a treatment plan can be formulated after a focused history, physical exam and basic lab-work are conducted. However, in certain complex cases, more testing can be employed. The major organic etiologies can be subdivided into vascular, neurologic, and endocrine. Specific testing should be directed by clinical clues noted during the preliminary evaluation. These tests vary in degree of invasiveness, precision, and at times may not affect treatment. Results should be integrated into the overall clinical picture to assist in diagnosis and help guide therapy.


Identification of GPC2 as an Oncoprotein and Candidate Immunotherapeutic Target in High-Risk Neuroblastoma.
We developed an RNA-sequencing-based pipeline to discover differentially expressed cell-surface molecules in neuroblastoma that meet criteria for optimal immunotherapeutic target safety and efficacy. Here, we show that GPC2 is a strong candidate immunotherapeutic target in this childhood cancer. We demonstrate high GPC2 expression in neuroblastoma due to MYCN transcriptional activation and/or somatic gain of the GPC2 locus. We confirm GPC2 to be highly expressed on most neuroblastomas, but not detectable at appreciable levels in normal childhood tissues. In addition, we demonstrate that GPC2 is required for neuroblastoma proliferation. Finally, we develop a GPC2-directed antibody-drug conjugate that is potently cytotoxic to GPC2-expressing neuroblastoma cells. Collectively, these findings validate GPC2 as a non-mutated neuroblastoma oncoprotein and candidate immunotherapeutic target.


Noise-induced hearing loss caused by gunshot in South Korean military service.
Noise-induced hearing loss is a preventive disease and yet the effective treatment modality has not been established. Acute acoustic traumas caused by an exposure to gunshot noise are common in young South Korean males in military service. Considering the significant lack of awareness on this serious issue as well as the absence of proper protective gear, an in-depth analysis is desperately needed. All 3650 soldiers performed regular periodic gunfire exercise without any hearing protective measures. Seven patients with hearing impairment after periodic gunfire visited the aeromedical squadron; all were right-handed males. Six were tested with the K-2 rifle and one was tested with a K-5 revolver. History taking, physical examination, pure-tone audiometry, and impedance audiometry were conducted. In the next periodic gunfire exercise, all 3650 soldiers performed gunfire with unilateral hearing protection. The average outcome of postexposure air conduction thresholds was 6.5 dB in the right ear and 33.1 dB in the left ear. After medical treatment, hearing impairment was much improved; however, tinnitus was not diminished. In the next periodic gunfire exercise, the result of a supplement of unilateral earplug protection proved its effectiveness on acoustic trauma caused by gunfire noise. Asymmetry in hearing loss is related to the position of the head during gunfire. A unilateral hearing protection device was enough to protect hearing from gunfire noise. At the same time, it can effectively prevent a potential firearm accident that can be caused by trainees mishearing the instruction of a firearm instructor if both earplugs are worn. Thus, providing a unilateral earplug for protection against acoustic trauma must be taken into serious consideration.


Complementary food supplements to achieve micronutrient adequacy for infants and young children.
Many children in developing countries survive on a nutritionally inadequate diet. Dietary inadequacies during the complementary feeding period can be prevented by using complementary food supplements (CFSs) such as water dispersible or crushable micronutrient tablets, micronutrient sprinkles added to food just before feeding, or fortified spreads added to food just before feeding or fed as a snacks. A meeting was convened to discuss technical and operational issues related to the development of these new approaches and to identify knowledge gaps. The technical issues covered: what micronutrients to include, tolerable upper intake limits, bioavailability, micronutrient and macronutrient stability, package systems and amounts, encapsulation technologies, methods to limit or eliminate allergens, bacterial and chemical contamination, interactions between CFSs and complementary foods, and flavoring agents. Operational issues included: identifying the market positioning of CFSs, cost positioning of CFSs, regulatory requirements, CFS production and technology transfer, quality assurance, and public-private sector partnership and coordination. Intervention trials are needed to determine the efficacy of CFSs in preventing micronutrient deficiencies. Other important knowledge gaps relate to technical and operational issues. Sprinkles and tablets are produced using well-known technologies, but further research is needed to modify them for use as CFSs. Spread development is not as advanced as sprinkle and tablet development, and further research is needed to improve the technology. Although none of the products is ready for widespread use, enough information is available to set research priorities and accelerate product development and implementation.


A high-sensitivity method for detection and measurement of HMGB1 protein concentration by high-affinity binding to DNA hemicatenanes.
Protein HMGB1, an abundant nuclear non-histone protein that interacts with DNA and has an architectural function in chromatin, was strikingly shown some years ago to also possess an extracellular function as an alarmin and a mediator of inflammation. This extracellular function has since been actively studied, both from a fundamental point of view and in relation to the involvement of HMGB1 in inflammatory diseases. A prerequisite for such studies is the ability to detect HMGB1 in blood or other biological fluids and to accurately measure its concentration. In addition to classical techniques (western blot, ELISA) that make use of specific anti-HMGB1 antibodies, we present here a new, extremely sensitive technique that is based on the fact that hemicatenated DNA loops (hcDNA) bind HMGB1 with extremely high affinity, higher than the affinity of specific antibodies, similar in that respect to DNA aptamers. DNA-protein complexes formed between HMGB1 and radiolabeled hcDNA are analyzed by electrophoresis on nondenaturing polyacrylamide gels using the band-shift assay method. In addition, using a simple and fast protocol to purify HMGB1 on the basis of its solubility in perchloric acid allowed us to increase the sensitivity by suppressing any nonspecific background. The technique can reliably detect HMGB1 at a concentration of 1 pg per microliter in complex fluids such as serum, and at much lower concentrations in less complex samples. It compares favorably with ELISA in terms of sensitivity and background, and is less prone to interference from masking proteins in serum. The new technique, which illustrates the potential of DNA nanoobjects and aptamers to form high-affinity complexes with selected proteins, should provide a valuable tool to further investigate the extracellular functions of HMGB1 and its involvement in inflammatory pathologies.


The effects of anatabine on non-invasive indicators of muscle damage: a randomized, double-blind, placebo-controlled, crossover study.
Anatabine (ANA), a minor tobacco alkaloid found in the Solanaceae family of plants, may exhibit anti-inflammatory activity, which may be useful to aid in recovery from exercise-induced muscle damage. The purpose of this study, therefore, was to examine the effects of ANA supplementation on the recovery of isometric strength and selected non-invasive indicators of muscle damage. A double-blinded, placebo-controlled, crossover design was used to study eighteen men (mean ± SD age = 22.2 ± 3.1 yrs; body mass = 80.3 ± 15.7 kg) who participated in two randomly-ordered conditions separated by a washout period. The ANA condition consisted of consuming 6-12 mg anatabine per day for 10 days, while testing took place during days 7-10. The placebo (PLA) condition was identical except that the PLA supplement contained no ANA. Maximal voluntary isometric peak torque (PT) of the forearm flexors, arm circumference, hanging joint angle, and subjective pain ratings were measured before (PRE), immediately after (POST), and 24, 48, and 72 h after six sets of 10 maximal, eccentric isokinetic forearm flexion muscle actions. Resting heart rate and blood pressure were measured at PRE and 72 h in each condition. For PT, hanging joint angle, arm circumference, and subjective pain ratings, there were no condition x time (p > 0.05) interactions, there were no main effects for condition (p > 0.05), but there were main effects for time (p < 0.001). There were no condition x time (p > 0.05) interactions and no main effects for condition (p > 0.05) or time (p > 0.05) for blood pressure or resting heart rate. ANA supplementation had no effect on the recovery of muscle strength, hanging joint angle, arm swelling, or subjective pain ratings after a bout of maximal eccentric exercise in the forearm flexors. Therefore, ANA may not be beneficial for those seeking to improve recovery from heavy eccentric exercise. Future studies should examine the effects of ANA on the pro-inflammatory cytokine responses to exercise-induced muscle damage and the chronic low-grade inflammation observed in obese and elderly individuals.


Mitochondrial Compartmentalization Confers Specificity to the 2-Ketoacid Recursive Pathway: Increasing Isopentanol Production in Saccharomyces cerevisiae.
Recursive elongation pathways produce compounds of increasing carbon-chain length with each iterative cycle. Of particular interest are 2-ketoacids derived from recursive elongation, which serve as precursors to a valuable class of advanced biofuels known as branched-chain higher alcohols (BCHAs). Protein engineering has been used to increase the number of iterative elongation cycles completed, yet specific production of longer-chain 2-ketoacids remains difficult to achieve. Here, we show that mitochondrial compartmentalization is an effective strategy to increase specificity of recursive pathways to favor longer-chain products. Using 2-ketoacid elongation as a proof of concept, we show that overexpression of the three elongation enzymes-LEU4, LEU1, and LEU2-in mitochondria of an isobutanol production strain results in a 2.3-fold increase in the isopentanol to isobutanol product ratio relative to overexpressing the same elongation enzymes in the cytosol, and a 31-fold increase relative to wild-type enzyme expression. Reducing the loss of intermediates allows us to further boost isopentanol production to 1.24 ± 0.06 g/L of isopentanol. In this strain, isopentanol accounts for 86% of the total BCHAs produced, while achieving the highest isopentanol titer reported for Saccharomyces cerevisiae. Localizing the elongation enzymes in mitochondria enables the development of strains in which isopentanol constitutes as much as 93% of BCHA production. This work establishes mitochondrial compartmentalization as a new approach to favor high titers and product specificities of larger products from recursive pathways.


Can long-term dentine bonding created in real life be forecasted by parameters established in the laboratory?
Tooth-coloured plastic dental fillings secured by adhesives to tooth structures are widely used to fix decayed teeth. Whereas laboratory tests demonstrate rapid deterioration of the ability of the adhesives to stick to dentine, clinical studies show that these fillings are relatively durable. This discrepancy suggests that the parameters used for simulating bond degradation in the laboratory setting do not correlate well with clinical outcomes. The present study examined the long-term tensile bond strength of resin composite fillings performed in real life and under different laboratory-simulated bonding conditions to identify parameters that may be used to forecast the durability of adhesive bonds created in dentine. Fillings placed in vivo were subjected to different periods of intraoral function. In vitro specimens were bonded based on whether simulated pulpal pressure (SPP) or thermomechanical cycling was implemented, and how long the completed fillings were stored in water. Thermomechanical cycling used in combination with long-term water ageing are useful in forecasting the decline in strength of resin-dentine bonds created in vivo. These parameters should be adopted for future evaluations. Conversely, the use of SPP does not appear to be a significant parameter in the simulation of long-term clinical deterioration of bond integrity.


The sex locus is tightly linked to factors conferring sex-specific lethal effects in the mosquito Aedes aegypti.
In many taxa, sex chromosomes are heteromorphic and largely non-recombining. Evolutionary models predict that spread of recombination suppression on the Y chromosome is fueled by the accumulation of sexually antagonistic alleles in close linkage to the sex determination region. However, empirical evidence for the existence of sexually antagonistic alleles is scarce. In the mosquito Aedes aegypti, the sex-determining chromosomes are homomorphic. The region of suppressed recombination, which surrounds the male-specific sex-determining gene, remains very small, despite ancient origin of the sex chromosomes in the Aedes lineage. We conducted a genetic analysis of the A. aegypti chromosome region tightly linked to the sex locus. We used a strain with an enhanced green fluorescent protein (EGFP)-tagged transgene inserted near the male-determining gene to monitor crossing-over events close to the boundary of the sex-determining region (SDR), and to trace the inheritance pattern of the transgene in relation to sex. In a series of crossing experiments involving individuals with a recombinant sex chromosome we found developmental abnormalities leading to 1:2 sex biases, caused by lethality of half of the male or female progeny. Our results suggest that various factors causing sex-specific lethal effects are clustered within the neighborhood of the SDR, which in the affected sex are likely lost or gained through recombination, leading to death. These may include genes that are recessive lethal, vital for development and/or sexually antagonistic. The sex chromosome fragment in question represents a fascinating test case for the analysis of processes that shape stable boundaries of a non-recombining region.


Role for DNA methylation in the regulation of miR-200c and miR-141 expression in normal and cancer cells.
The microRNA-200 family participates in the maintenance of an epithelial phenotype and loss of its expression can result in epithelial to mesenchymal transition (EMT). Furthermore, the loss of expression of miR-200 family members is linked to an aggressive cancer phenotype. Regulation of the miR-200 family expression in normal and cancer cells is not fully understood. Epigenetic mechanisms participate in the control of miR-200c and miR-141 expression in both normal and cancer cells. A CpG island near the predicted mir-200c/mir-141 transcription start site shows a striking correlation between miR-200c and miR-141 expression and DNA methylation in both normal and cancer cells, as determined by MassARRAY technology. The CpG island is unmethylated in human miR-200/miR-141 expressing epithelial cells and in miR-200c/miR-141 positive tumor cells. The CpG island is heavily methylated in human miR-200c/miR-141 negative fibroblasts and miR-200c/miR-141 negative tumor cells. Mouse cells show a similar inverse correlation between DNA methylation and miR-200c expression. Enrichment of permissive histone modifications, H3 acetylation and H3K4 trimethylation, is seen in normal miR-200c/miR-141-positive epithelial cells, as determined by chromatin immunoprecipitation coupled to real-time PCR. In contrast, repressive H3K9 dimethylation marks are present in normal miR-200c/miR-141-negative fibroblasts and miR-200c/miR-141 negative cancer cells and the permissive histone modifications are absent. The epigenetic modifier drug, 5-aza-2'-deoxycytidine, reactivates miR-200c/miR-141 expression showing that epigenetic mechanisms play a functional role in their transcriptional control. We report that DNA methylation plays a role in the normal cell type-specific expression of miR-200c and miR-141 and this role appears evolutionarily conserved, since similar results were obtained in mouse. Aberrant DNA methylation of the miR-200c/141 CpG island is closely linked to their inappropriate silencing in cancer cells. Since the miR-200c cluster plays a significant role in EMT, our results suggest an important role for DNA methylation in the control of phenotypic conversions in normal cells.


The kidney as a reservoir for HIV-1 after renal transplantation.
Since the recent publication of data showing favorable outcomes for patients with HIV-1 and ESRD, kidney transplantation has become a therapeutic option in this population. However, reports have documented unexplained reduced allograft survival in these patients. We hypothesized that the unrecognized infection of the transplanted kidney by HIV-1 can compromise long-term allograft function. Using electron microscopy and molecular biology, we examined protocol renal transplant biopsies from 19 recipients with HIV-1 who did not have detectable levels of plasma HIV-1 RNA at transplantation. We found that HIV-1 infected the kidney allograft in 68% of these patients. Notably, HIV-1 infection was detected in either podocytes predominately (38% of recipients) or tubular cells only (62% of recipients). Podocyte infection associated with podocyte apoptosis and loss of differentiation markers as well as a faster decline in allograft function compared with tubular cell infection. In allografts with tubular cell infection, epithelial cells of the proximal convoluted tubules frequently contained abnormal mitochondria, and both patients who developed features of subclinical acute cellular rejection had allografts with tubular cell infection. Finally, we provide a novel noninvasive test for determining HIV-1 infection of the kidney allograft by measuring HIV-1 DNA and RNA levels in patients' urine. In conclusion, HIV-1 can infect kidney allografts after transplantation despite undetectable viremia, and this infection might influence graft outcome.


Infarct characterization using CT.
Myocardial infarction (MI) is a major cause of death and disability worldwide. The incidence is not expected to diminish, despite better prevention, diagnosis and treatment, because of the ageing population in industrialized countries and unhealthy lifestyles in developing countries. Nowadays it is highly requested an imaging tool able to evaluate MI and viability. Technology improvements determined an expansion of clinical indications from coronary plaque evaluation to functional applications (perfusion, ischemia and viability after MI) integrating additional phases and information in the mainstream examination. Cardiac computed tomography (CCT) and cardiac MR (CMR) employ different contrast media, but may characterize MI with overlapping imaging findings due to the similar kinetics and tissue distribution of gadolinium and iodinated contrast media. CCT may detect first-pass perfusion defects, dynamic perfusion after pharmacological stress, and delayed enhancement (DE) of non-viable territories.


Molecular Docking Study Based on Pharmacophore Modeling for Novel PhosphodiesteraseIV Inhibitors.
In this study, pharmacophore modelling was carried out for novel PhosphodiesteraseIV (PDEIV) inhibitors. A pharmacophore-based virtual screening, which resulted in 1959 hit compounds was performed with six chemical databases. The pharmacophore screening was proven to be successful in discriminating active and inactive inhibitors using a set of compounds with known activity obtained from ChEMBL database. Furthermore, the Lipinski's rule of five was applied for physicochemical filtering of the hit molecules and this yielded 1840 compounds. Three docking software tools, AutoDock 4.0, AutoDock Vina, and Gold v5.1 were used for the docking process. All 1840 compounds and the known selective inhibitor, rolipram, were docked into the active site of the target protein. A total of 234 compounds with all three scoring values higher than those of rolipram were determined with the three docking tools. The interaction maps of 14 potent inhibitors complexed with PDEIV B and D isoforms have been analyzed and seven key residues (Asn 395, Gln 443, Tyr 233, Ile 410, Phe 446, Asp 392, Thr 407) were found to interact with more than 80 % of the potent inhibitors. For each one of the 234 hit compounds, using the bound conformation with the highest AutoDock score, the interacting residues were determined. 117 out of 234 compounds are found to interact with at least five of the seven key residues and these were selected for further evaluation. The conformation with the highest AutoDock score for each 117 compounds were rescored using the DSX scoring function. This yielded a total of 101 compounds with better score values than the natural ligand rolipram. For ADME/TOX calculations, the FAF-Drugs2 server was used and 32 out of 101 compounds were found to be non-toxic.


The dynamics of genome replication using deep sequencing.
Eukaryotic genomes are replicated from multiple DNA replication origins. We present complementary deep sequencing approaches to measure origin location and activity in Saccharomyces cerevisiae. Measuring the increase in DNA copy number during a synchronous S-phase allowed the precise determination of genome replication. To map origin locations, replication forks were stalled close to their initiation sites; therefore, copy number enrichment was limited to origins. Replication timing profiles were generated from asynchronous cultures using fluorescence-activated cell sorting. Applying this technique we show that the replication profiles of haploid and diploid cells are indistinguishable, indicating that both cell types use the same cohort of origins with the same activities. Finally, increasing sequencing depth allowed the direct measure of replication dynamics from an exponentially growing culture. This is the first time this approach, called marker frequency analysis, has been successfully applied to a eukaryote. These data provide a high-resolution resource and methodological framework for studying genome biology.


Pathways Governing Polyethylenimine Polyplex Transfection in Microporous Annealed Particle Scaffolds.
Gene delivery using injectable hydrogels can serve as a potential method for regulated tissue regeneration in wound healing. Our microporous annealed particle (MAP) hydrogel has been shown to promote cellular infiltration in both skin and brain wounds, while reducing inflammation. Although the scaffold itself can promote healing, it is likely that other signals will be required to promote healing of hard-to-treat wounds. Gene delivery is one approach to introduce desired bioactive signals. In this study, we investigated how the properties of MAP hydrogels influence non-viral gene delivery of polyethylenimine-condensed plasmid to cells seeded within the MAP gel. From past studies, we found that gene transfer to cells seeded in tissue culture plastic differed from gene transfer to cells seeded inside hydrogel scaffolds. Since MAP scaffolds are generated from hydrogel microparticles that are approximately 100 μm in diameter, they display local characteristics that can be viewed as two-dimensional or three-dimensional to cells. Thus, we sought to study if gene transfer inside MAP scaffolds differed from gene transfer to cells seeded in tissue culture plastic. We sought to understand the roles of the endocytosis pathway, actin and microtubule dynamics, RhoGTPases, and YAP/TAZ on transfection of human fibroblasts.


Dual adaptive dynamic control of mobile robots using neural networks.
This paper proposes two novel dual adaptive neural control schemes for the dynamic control of nonholonomic mobile robots. The two schemes are developed in discrete time, and the robot's nonlinear dynamic functions are assumed to be unknown. Gaussian radial basis function and sigmoidal multilayer perceptron neural networks are used for function approximation. In each scheme, the unknown network parameters are estimated stochastically in real time, and no preliminary offline neural network training is used. In contrast to other adaptive techniques hitherto proposed in the literature on mobile robots, the dual control laws presented in this paper do not rely on the heuristic certainty equivalence property but account for the uncertainty in the estimates. This results in a major improvement in tracking performance, despite the plant uncertainty and unmodeled dynamics. Monte Carlo simulation and statistical hypothesis testing are used to illustrate the effectiveness of the two proposed stochastic controllers as applied to the trajectory-tracking problem of a differentially driven wheeled mobile robot.


Estimation of the burden of active and life-time epilepsy: a meta-analytic approach.
To estimate the burden of lifetime epilepsy (LTE) and active epilepsy (AE) and examine the influence of study characteristics on prevalence estimates. We searched online databases and identified articles using prespecified criteria. Random-effects meta-analyses were used to estimate the median prevalence in developed countries and in urban and rural settings in developing countries. The impact of study characteristics on prevalence estimates was determined using meta-regression models. The median LTE prevalence for developed countries was 5.8 per 1,000 (5th-95th percentile range 2.7-12.4) compared to 15.4 per 1,000 (4.8-49.6) for rural and 10.3 (2.8-37.7) for urban studies in developing countries. The median prevalence of AE was 4.9 per 1,000 (2.3-10.3) for developed countries and 12.7 per 1,000 (3.5-45.5) and 5.9 (3.4-10.2) in rural and urban studies in developing countries. The estimates of burden for LTE and AE in developed countries were 6.8 million (5th-95th percentile range 3.2-14.7) and 5.7 million (2.7-12.2), respectively. In developing countries these were 45 (14-145) million LTE and 17 (10-133) million AE in rural areas and 17 (5-61) million LTE and 10 (5-17) million AE in urban areas. Studies involving all ages or only adults showed higher estimates than pediatric studies. Higher prevalence estimates were also associated with rural location and small study size. This study estimates the global burden of epilepsy and the proportions with AE, which may benefit from treatment. There are systematic differences in reported prevalence estimates, which are only partially explained by study characteristics.


Endolymphatic sac shunt, labyrinthectomy, and vestibular nerve section in Meniere's disease.
Medical treatment for Meniere's disease is effective in controlling vertigo for approximately 85% of patients. However, when disabling vertigo continues, surgical therapy is indicated. Several surgical approaches are performed to control the symptoms of peripheral vestibular disorders refractory to medical measures, each procedure having many technical variations. Surgery is usually reserved for patients with disabling vertigo. Here, the authors discuss surgical options for vertigo control in Meniere's disease and review the literature on outcomes of these management options. The authors discuss endolymphatic sac shunt (ie, endolymphatic mastoid shunt), vestibular nerve section, cochleosacculotomy, and labyrinthectomy. When looking at data based on patient ratings, the authors find that surgery improves vertigo in endolymphatic sac shunt, vestibular nerve section, and labyrinthectomy groups and improves imbalance for the endolymphatic sac shunt and vestibular nerve section groups. Labyrinthectomy and translabyrinthine vestibular nerve section both offer excellent control of intractable vertigo. However, patients undergoing translab yrinthine vestibular nerve section are more likely to show improvement in imbalance and functional disability. This outcome is more likely for diagnoses other than Meniere's disease. There are potential prognostic factors that can be helpful in the preoperative or postoperative counseling of patients undergoing surgical treatment of vertigo. Patients who rate themselves as more disabled before surgery are less likely to achieve the best outcomes. Several other factors, such as duration of disease, contralateral tinnitus, eye disease, and allergy, may play a role.


Lysis of oncornaviruses by human serum. Isolation of the viral complement (C1) receptor and identification as p15E.
Moloney leukemia virus activated both the classical and alternative pathways of human complement. About 500,000 virions were required to detect activation of the classical pathway whereas 5,000 times as many virions were necessary to initiate the alternative pathway, indicating that in this system only the former is of biological significance. Disruption of the virus with Triton X-100 destroyed its ability to initiate the alternative pathway without affecting its ability to activate the classical pathway. After ultracentrifugation of disrupted virus the active component could be recovered in the supernate and was isolated by isoelectric focusing in granulated gels. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic and analysis and cyanogen bromide digestion studies revealed that the activity resided in a methionine-containing protein having a pI of 7.5 and a molecular weight of approximately equal to 15,000 daltons. The purified protein interacts strongly with Clq and efficiently activates Cl. RNase and lipolytic enzymes had no effect on the isolated protein but incubation with trypsin resulted in loss of activity. Enzymatic digestion studies of surface-labeled virus indicate that the active protein is a viral membrane protein. On the basis of these results it is concluded that the complement receptor of Moloney leukemia virus is the surface protein p15E.


Does payment drive procedures? Payment for specialty services and procedure rate variations in 3 HMOs.
To study how payment for specialty services affects the rates of performance of invasive procedures by physicians in a number of specialties. Observational study. Administrative data from 1996-1997 and 1997-1998 from 3 large health maintenance organizations (HMOs) in the Midwestern and western United States were used to study variations in procedure rates associated with different methods of paying for cardiology, gastroenterology, ophthalmology, orthopedic, and ear, nose and throat services within each HMO. The age-, sex-, and comorbidity adjusted probabilities of undergoing selected, potentially discretionary procedures, were compared within each plan by payment method. After adjustment, rates under fee-for-service payment tended to be higher than those under capitation or salary payment, whereas there was no clear pattern for salary versus capitation payment. Even within a single specialty in a single plan, however, rates did not always follow the same pattern for different procedures. The payment method for specialty services used by these 3 health plans was variably associated with how likely patients were to undergo a variety of invasive procedures. The effects of contract payment methods for specialty services on health care costs, quality, and outcomes should be further studied, but such studies will challenge the capabilities of health plan data systems.


Comparative Analysis of Urine Fractions for Optimal Bladder Cancer Detection Using DNA Methylation Markers.
DNA methylation analysis of full void urine and urine pellet seems promising for bladder cancer (BC) detection and surveillance. Urinary cell-free DNA from urine supernatant is now gaining interest for other molecular tests in BC. This study aims to evaluate which urine fraction is preferred for BC diagnosis using methylation markers: full void urine, urine pellet or supernatant. Methylation levels of nine markers were determined in the three urine fractions and correlated with their respective tumor tissues in BC patients and compared to controls. For all markers and marker panel GHSR/MAL, diagnostic performance was determined by calculating the area under the curve (AUC) of the respective receiver operating characteristic curves. For most of the markers, there was a significant correlation between the methylation levels in each of the urine fractions and the matched tumor tissues. Urine pellet was the most representative fraction. Generally, AUCs for BC diagnosis were comparable among the fractions. The highest AUC was obtained for GHSR/MAL in urine pellet: AUC 0.87 (95% confidence interval: 0.73-1.00), corresponding to a sensitivity of 78.6% and a specificity of 91.7%. Our results demonstrate that cellular and cell-free DNA in urine can be used for BC diagnosis by urinary methylation analysis. Based on our comparative analysis and for practical reasons, we recommend the use of urine pellet.


Investigation of clinically important benefit of anterior cervical decompression and fusion.
The objectives of the prospective randomized study are to investigate the clinically relevant change after anterior cervical decompression and fusion (ACDF) using measures of pain intensity (visual analog scale, VAS) and neck disability index (NDI). And to determine the number of subjects showing persistent pain and disability at 6-year follow-up. To investigate the possibility of differences in outcome between ACDF with the cervical intervertebral fusion cage (CIFC) and the Cloward procedure (CP). Clinically relevant change and residual, postoperative pain intensity and disability after ACDF have been investigated a little. Ninety-five patients with neck and radicular arm pain lasting for at least 6 months were randomly selected to receive ACDF with the CP or the CIFC. Questionnaires concerning pain and NDI were obtained from 83 patients (87%) at a mean follow-up time of 76 months (range 56-94 months). When evaluating clinical benefits regarding pain intensity 6 years after ACDF, according to different cut-off points and relative percentages, symptoms improved in 46-78% of patients. Improvement in NDI was seen in 18-20% of patients. Approximately 70% of the patients had persistent pain and disability at 6-year follow-up. There was no clinically important difference following CP versus CIFC. Thirty millimeter and 20% in pain intensity and NDI, respectively, are reasonable criteria to suggest a clinically relevant change after ACDF. Before patients undergo ACDF, they should be informed that they have an approximate 50% probability of achieving pain relief and little probability of functional improvement. The findings demonstrate that there is poor evidence for difference between CIFC and CP.


Effect of folic acid and vitamins B6 and B12 on microcirculatory vasoreactivity in patients with hyperhomocysteinemia.
Hyperhomocysteinemia (HHcy) has been identified as an independent risk factor for atherosclerotic vascular disease. The effect of high-dose folic acid or combination vitamin therapy for the treatment of HHcy on the microcirculation is unknown. The purpose of this study was to evaluate the effect of a combination of folic acid, vitamin B6, and vitamin B12 on endothelium-dependent and endothelium-independent vasoreactivity in patientswith HHcy. Baseline cutaneous microvascular vasoreactivity was measured in 20 patients with HHcy and 18 patients with normohomocysteinemia (NHcy). Laser Doppler scan imaging before and after iontophoresis of 1% acetylcholine chloride (endothelium-dependent response) and 1% sodium nitroprusside (endothelium-independent response) was performed for the measurement of forearm skin vasodilatation. Patients were then treated with 10 mg folic acid, 100 mg vitamin B6, and 1 mg vitamin B12 orally once a day for 6 months. Follow-up fasting serum homocysteine and cutaneous Laser Doppler scan imaging before and after iontophoresis were performed at 1, 2, 3, and 6 months. Statistical analysis was performed using Fisher's exact test, paired t test, and Wilcoxon matched-pairs signed-ranks test, with significance set at P < .05. The HHcy group was older than the NHcy group (70.89 +/- 1.95 vs 61.78 +/- 2.73 years, P = .02). Otherwise the groups were similar in terms of race, tobacco use, comorbid diseases, and serum lipoproteins. Over the 6-month period, fasting serum homocysteine levels decreased significantly in both the NHcy group (10.40 +/- 0.59 micromol/L vs 8.97 +/- 0.84 micromol/L, P = .01) and the HHcy group (19.80 +/- 1.06 micromol/L vs 13.40 +/- 0.86 micromol/L, P = .0002). There were no statistically significant changes in endothelium-independent vasoreactivity (voltage change from baseline) in either group. Endothelium-independent vasore activity decreased over the 6-month period in the HHcy group (0.20 +/- 0.04 V vs 0.11 +/- 0.03 V, P = .03). Subanalysis of HHcy with diabetes or age greater than 65 years both showed worsening trends in endothelium-independent vasoreactivity (P = .05 for both groups). There were no statistically significant changes in endothelium-independent vasoreactivity in the NHcy group. High doses of folic acid and vitamins B6 and B12 lower fasting serum homocysteine levels in patients with HHcy. Older and diabetic patients with HHcy tend to do worse possibly because of long-term fixed microvascular insult secondary to multiple sustained comorbidities.


Human papillomavirus, smoking status and outcomes in tonsillar squamous cell carcinoma.
It is now clear that the two separate entitles of tonsillar cancer, HPV induced and non-HPV induced (smoking induced), have significantly different presenting stage and outcomes. A significant proportion of patients with human papillomavirus positive tonsillar cancer have had exposure to smoking. We examined the combined effect of human papillomavirus and smoking on the outcomes and determined whether smoking can modify the beneficial effect of human papillomavirus. A total of 403 patients from nine centers were followed up for recurrence or death for a median of 38 months. Determinants of the rate of loco-regional recurrence, death from tonsillar cancer and overall survival were modeled using Cox regression. Smoking status was a significant predictor of overall survival (p = 0.04). There were nonstatistically significant trends favoring never smokers for loco-regional recurrence and disease specific survival. In addition, there was no statistically significant interactions between smoking and human papillomavirus (p-values for the interaction were 0.26 for loco-regional recurrence, 0.97 for disease specific survival and 0.73 for overall survival). The effect of smoking on loco-regional recurrence and disease specific survival outcomes was not statistically significant, nor was there significant evidence that the effect of smoking status on these outcomes was modified by HPV status. Irrespective of HPV status, however, smokers did have poorer overall survival than never-smokers, presumably due to effects of smoking that are unrelated to the primary cancer.


Dynamic multimodal interactions in navigating wood ants: what do path details tell us about cue integration?
Ants are expert navigators, using multimodal information to navigate successfully. Here, we present the results of systematic studies of multimodal cue use in navigating wood ants, Formica rufa Ants learnt to navigate to a feeder that was defined by an olfactory cue (O), visual cue (V) and airflow (A) presented together. When the feeder, olfactory cue and airflow were all placed at the centre of the visual cue (VOACentre), ants did not directly approach the learnt feeder when either the olfactory or visual cue was removed. This confirms that some form of cue binding has taken place. However, in a visually simpler task with the feeder located at the edge of the visual cue (VOAEdge), ants still approached the feeder directly when individual cue components were removed. Hence, cue binding is flexible and depends on the navigational context. In general, cues act additively in determining the ants' path accuracy, i.e. the use of multiple cues increased navigation performance. Moreover, across different training conditions, we saw different motor patterns in response to different sensory cues. For instance, ants had more sinuous paths with more turns when they followed an odour plume but did not have any visual cues. Having visual information together with the odour enhanced performance and therefore positively impacted on plume following. Interestingly, path characteristics of ants from the different multimodal groups (VOACentre versus VOAEdge) were different, suggesting that the observed flexibility in cue binding may be a result of ants' movement characteristics.


Clinical research ethics for critically ill patients: a pandemic proposal.
Pandemic H1N1 influenza is projected to be unprecedented in its scope, causing acute critical illness among thousands of young otherwise healthy adults, who will need advanced life support. Rigorous, relevant, timely, and ethical clinical and health services research is crucial to improve their care and outcomes. Studies designed and conducted during a pandemic should be held to the same high methodologic and implementation standards as during other times. However, unique challenges arise with the need to conduct investigations as efficiently as possible, focused on the optimal outcome for the individual patient, while balancing the need for maximal societal benefit. We believe that clinical critical care research during a pandemic must be approached differently from research undertaken under nonemergent circumstances. We propose recommendations to clinical investigators and research ethics committees regarding clinical and health services research on pandemic-related critical illness. We also propose strategies such as expedited and centralized research ethics committee reviews and alternate consent models.


Does mutational robustness inhibit extinction by lethal mutagenesis in viral populations?
Lethal mutagenesis is a promising new antiviral therapy that kills a virus by raising its mutation rate. One potential shortcoming of lethal mutagenesis is that viruses may resist the treatment by evolving genomes with increased robustness to mutations. Here, we investigate to what extent mutational robustness can inhibit extinction by lethal mutagenesis in viruses, using both simple toy models and more biophysically realistic models based on RNA secondary-structure folding. We show that although the evolution of greater robustness may be promoted by increasing the mutation rate of a viral population, such evolution is unlikely to greatly increase the mutation rate required for certain extinction. Using an analytic multi-type branching process model, we investigate whether the evolution of robustness can be relevant on the time scales on which extinction takes place. We find that the evolution of robustness matters only when initial viral population sizes are small and deleterious mutation rates are only slightly above the level at which extinction can occur. The stochastic calculations are in good agreement with simulations of self-replicating RNA sequences that have to fold into a specific secondary structure to reproduce. We conclude that the evolution of mutational robustness is in most cases unlikely to prevent the extinction of viruses by lethal mutagenesis.


Exercise and the breast cancer survivor: the role of the nurse practitioner.
Patients with cancer are living longer with their disease and have improved survival rates because of early detection and more effective cancer treatments. Lifestyle modification and exercise improve clinical outcomes in breast cancer survivors. Exercise has important implications for the survivor and should be integrated into the aftercare trajectory of survivorship. A literature review of articles published from 2002-2014 was conducted using the key words cancer survivor, survivorship, breast cancer, collaboration, and exercise. PubMed, Cochrane Database of Systematic Reviews, and CINAHL® databases were searched. Nurse practitioners can build an environment to motivate patients to exercise, empowering them to be active participants in their own survivorship care. Collaboration is necessary to ensure that healthy lifestyle choices, including exercise, are being discussed and implemented in survivorship care plans to help optimize patient outcomes.


Lipoprotein transport in the blood stream to the arterial wall in a carotid artery bifurcation.
The carotid arteries are a common site of atherosclerotic plaque formation, which has been linked to the blood flow patterns and the mass transport phenomenon. The purpose of this research was to study the lipid transport in a human carotid artery model, focusing on the effects of local geometric and hemodynamic factors on mass transfer from blood flow to vessel wall and its concentration at the luminal surface of the artery. The Reynolds number, 250, and the Schmidt number, 6.66x105, were selected to model the mass transfer of LDL macro molecules, and in order to see the effect of Reynolds and Schmidt numbers to mass transport, the model was analyzed with different conditions. The steady state flow was used for two dimensional carotid geometry. At the inlet, the blood flow was assumed a steady fully developed laminar velocity profile with a uniform LDL concentration. The vessel wall was assumed permeable to water and semi-permeable to LDL macro molecules. The problem was analyzed with the finite element method. The results show 26% increase of LDL concentration from inlet value at the luminal surface of the artery located in the separated flow region. The maximum value of LDL concentration occurred at the separation point.


Competitive binding for triggering a fluorescence response in a hydrazodicarboxamide-based [2]rotaxane.
The design and synthesis of an interlocked receptor based on a hydrogen bonded [2]rotaxane containing a hydrazodicarboxamide binding site are reported. An anion recognition process with tetrabutylammonium benzoate triggers the submolecular translational movement of its benzylic amide macrocycle developing a progressive increase of the fluorescence intensity, effectively quenched at the original state. The binding of the anion competes with the hydrogen-bond-connected cyclic component for the bis(urea)-based station pushing it towards a stoppered alkyl chain. Moreover, this interlocked system is able to work as a molecular switch restoring its initial state in two ways, either by an ion exchange reaction or by a high yielding oxidation/reduction sequence.


Segmental duplication of the basilar artery with thrombosis.
Duplication or fenestration of the basilar artery, a result of an embryologic malformation, has an incidence of up to 5.3% in the general population. The most common complication of this anomaly is the formation of aneurysms. Thrombosis of a partially duplicated basilar artery developed in a 43-year-old man who complained of visual disturbances followed by seizures and coma, and who eventually died. Autopsy showed a partially organized thrombus occluding both halves of a duplicated portion of the basilar artery, old infarcts in the calcarine cortices, and a recent large infarct in the basis pontis. There was only minimal atherosclerosis of other intracranial arteries, including the vertebral arteries. Hemodynamic disturbances and turbulent blood flow at the site of fenestration may be the cause of the thrombosis that occurred in this artery.


Validation and Invalidation of Chemical Probes for the Human N-myristoyltransferases.
On-target, cell-active chemical probes are of fundamental importance in chemical and cell biology, whereas poorly characterized probes often lead to invalid conclusions. Human N-myristoyltransferase (NMT) has attracted increasing interest as target in cancer and infectious diseases. Here we report an in-depth comparison of five compounds widely applied as human NMT inhibitors, using a combination of quantitative whole-proteome N-myristoylation profiling, biochemical enzyme assays, cytotoxicity, in-cell protein synthesis, and cell-cycle assays. We find that N-myristoylation is unaffected by 2-hydroxymyristic acid (100 μM), D-NMAPPD (30 μM), or Tris-DBA palladium (10 μM), with the latter compounds causing cytotoxicity through mechanisms unrelated to NMT. In contrast, drug-like inhibitors IMP-366 (DDD85646) and IMP-1088 delivered complete and specific inhibition of N-myristoylation in a range of cell lines at 1 μM and 100 nM, respectively. This study enables the selection of appropriate on-target probes for future studies and suggests the need for reassessment of previous studies that used off-target compounds.


Phynx: an open source software solution supporting data management and web-based patient-level data review for drug safety studies in the general practice research database and other health care databases.
To develop a software solution that supports management and clinical review of patient data from electronic medical records databases or claims databases for pharmacoepidemiological drug safety studies. We used open source software to build a data management system and an internet application with a Flex client on a Java application server with a MySQL database backend. The application is hosted on Amazon Elastic Compute Cloud. This solution named Phynx supports data management, Web-based display of electronic patient information, and interactive review of patient-level information in the individual clinical context. This system was applied to a dataset from the UK General Practice Research Database (GPRD). Our solution can be setup and customized with limited programming resources, and there is almost no extra cost for software. Access times are short, the displayed information is structured in chronological order and visually attractive, and selected information such as drug exposure can be blinded. External experts can review patient profiles and save evaluations and comments via a common Web browser. Phynx provides a flexible and economical solution for patient-level review of electronic medical information from databases considering the individual clinical context. It can therefore make an important contribution to an efficient validation of outcome assessment in drug safety database studies.


Preparation and characterization of simvastatin/DMβCD complex and its pharmacokinetics in rats.
Simvastatin is poorly bioavailable because it is practically insoluble in water and shows dissolution rate-limited absorption. Solubilizing effects of several β-cyclodextrin (βCD) derivatives such as HPβCD, SBEβCD and DMβCD on simvastatin in aqueous solution were investigated using the phase solubility technique. The solubility diagram of simvastatin with each βCD derivative could be classified as AL-type, indicating soluble complex formation of 1:1 stoichiometry. Among the above βCD derivatives DMβCD was found to be the ideal complexing agent for improving drug solubility. The simvastatin complex with DMβCD was prepared using the co-evaporation method and was then characterized by differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FT-IR) and in vitro dissolution. Dissolution and pharmacokinetic studies indicated that the simvastatin/DMβCD complex exhibited an increased dissolution rate, rapid absorption, and improved bioavailability in rats compared to free drug. Maximum plasma concentration (cmax) and the time to reach it (tmax) were 21.86 μg mL-1 and 1.4 h for the drug complex, 8.25 μg mL-1 and 3.0 h for free drug, respectively. Main pharmacokinetic parameters such as tmax, cmax were significantly different (p < 0.01) between the simvastatin complex and free drug. Bioavailability of the simvastatin complex relative to free drug was up to 167.0 %.


Self-assembled nanoparticle micro-shells templated by liquid crystal sorting.
A current goal in nanotechnology focuses on the assembly of different nanoparticle types into 3D organized structures. In this paper we report the use of a liquid crystal host phase in a new process for the generation of micron-scale vesicle-like nanoparticle shells stabilized by ligand-ligand interactions. The constructs formed consist of a robust, thin spherical layer, composed of closely packed quantum dots (QDs) and stabilized by local crystallization of the mesogenic ligands. Ligand structure can be tuned to vary QD packing within the shell and made UV cross-linkable to allow for intact shell extraction into toluene. The assembly method we describe could be extended to other nanoparticle types (metallic, magnetic etc.), where hollow shell formation is controlled by thermally sorting mesogen-functionalized nanoparticles in a liquid crystalline host material at the isotropic to nematic transition. This process represents a versatile method for making non-planar 3D nano-assemblies.


Eye and Vision Health for Tomorrow: From Recommendations to Coordinated Action.
In 2016, the report Making Eye Health a Population Health Imperative: Vision for Tomorrow was published by the National Academies of Sciences, Engineering, and Medicine. Conference presentations and key stakeholder discussions have continued the dialogue initiated by this report. In 2018, a stakeholder meeting assessed the feasibility of establishing a backbone organization, a national center that can provide technical and resource assistance regarding eye and vision health. This meeting also resulted in a consensus to translate the report recommendations into action through 7 core functions: (1) supporting ongoing surveillance efforts, (2) establishing and strengthening a national multisector network of partner organizations, (3) creating an online vision and population health toolkit, (4) integrating and expanding early-detection initiatives across diverse clinical and nonclinical settings, (5) expanding eye and vision health education, (6) identifying and disseminating evidence-based interventions, and (7) exploring the integration of eye and vision health interventions into broader chronic care models. The report's recommendations aim to improve vision and health equity in the United States and should be implemented through an ongoing centrally coordinated campaign.


Views of women and men in Bobo-Dioulasso, Burkina Faso, on three forms of female genital modification.
This paper is about how female circumcision/female genital mutilation (FC/FGM) was viewed by women and men aged 18-89 in Bobo-Dioulasso, Burkina Faso, now that it has been against the law for almost 15 years. The findings come from 11 months of field research, participant observation and interviews in 2008. The practice of FC/FGM was an important issue in Bobo-Dioulasso, even though prevalence seems to be falling. The most important argument for continuing it was not a traditional role, but the need to control female sexuality - regarded as very active - not to negate it, but to ensure morally acceptable behaviour. When I talked about female genital cosmetic surgery it emerged that Bobolaise women used various substances to enhance sexual pleasure for men, both to keep the relationship and to protect the gifts and money many women needed to survive and for their children. FC/FGM was seen as a socio-cultural obligation, necessary to achieve a respectable status. Other forms of genital modification were seen as a means of satisfying male sexual needs, though vaginal tightening to hide sexual experience was also a way of demonstrating respectability. What emerged overall is that Bobolaises had their own perspectives about all the forms of female genital modification that were discussed.


Outcome after percutaneous transluminal angioplasty of arterial stenosis in renal transplant patients.
Ensuring graft survival in renal transplant patients is of paramount importance. Early detection and treatment of complications such as transplant renal artery stenosis (TRAS) are essential. To evaluate the technical and clinical success rate of renal transplant patients with stenosis in the transplant renal artery or in the iliac artery after percutaneous transluminal angioplasty (PTA). PTA was carried out on 24 patients with TRAS or iliac artery stenosis. Altogether, 28 stenoses were treated with PTA. The immediate technical result and the clinical outcomes after 1 and 3 months were assessed as well as clinical adverse events. A reduction in serum creatinine and/or a reduction in the number of antihypertensive drugs were criteria for clinical success. The immediate technical success rate after PTA was 93%. The clinical success rate after 1 month was 58%, increasing to 75% after 3 months. The technical success rate is not equivalent to the clinical success rate when treating TRAS with PTA. Furthermore, there is a delay in clinical response, sometimes of 3 months, after a technically successful PTA.


Kettlebell Training for Female Ballet Dancers: Effects on Lower Limb Power and Body Balance.
The aim of the present study was to evaluate the effects of 5 month kettlebell-based training on jumping performance, balance, blood pressure and heart rate in female classical ballet dancers. It was a clinical trial study with 23 female dancers (age = 21.74 ± 3.1 years; body height = 168.22 ± 5.12 cm; body mass = 53.69 ± 5.91 kg) took part in the study. Participants were divided into two groups: a kettlebell group (n = 13), that followed a commercial kettlebell training protocol named the "Simple & Sinister protocol", and a traditional dance training control group (n = 10). In the kettlebell group, kettlebell training completely replaced the jump and balance section of dance classes. Both groups performed balance and jumping tests before and after the training period. Blood pressure and the heart rate were also measured. The kettlebell group showed significant improvements in the balance tests (antero-posterior and medio-lateral oscillation) with both legs and eyes open as well as in all types of jump exercises (unrotated: +39.13%, p < 0.005; with a turnout: +53.15%, p < 0.005), while maximum and minimum blood pressure and the heart rate decreased significantly (max: -7.90%, p < 0.05; min: -9.86%, p < 0.05; Heart rate: -17.07%, p < 0.01). The results for the control group were non-significant for any variable. Comparison between groups showed significant differences for all variables analyzed, with greater improvements for the kettlebell group. Our results suggest that specific kettlebell training could be effective in improving jump performance and balance in classical dancers to a significantly greater degree compared to classical dance training.


Deciphering serology to understand the ecology of infectious diseases in wildlife.
The ecology of infectious disease in wildlife has become a pivotal theme in animal and public health. Studies of infectious disease ecology rely on robust surveillance of pathogens in reservoir hosts, often based on serology, which is the detection of specific antibodies in the blood and is used to infer infection history. However, serological data can be inaccurate for inference to infection history for a variety of reasons. Two major aspects in any serological test can substantially impact results and interpretation of antibody prevalence data: cross-reactivity and cut-off thresholds used to discriminate positive and negative reactions. Given the ubiquitous use of serology as a tool for surveillance and epidemiological modeling of wildlife diseases, it is imperative to consider the strengths and limitations of serological test methodologies and interpretation of results, particularly when using data that may affect management and policy for the prevention and control of infectious diseases in wildlife. Greater consideration of population age structure and cohort representation, serological test suitability and standardized sample collection protocols can ensure that reliable data are obtained for downstream modeling applications to characterize, and evaluate interventions for, wildlife disease systems.


Statistical emulation of landslide-induced tsunamis at the Rockall Bank, NE Atlantic.
Statistical methods constitute a useful approach to understand and quantify the uncertainty that governs complex tsunami mechanisms. Numerical experiments may often have a high computational cost. This forms a limiting factor for performing uncertainty and sensitivity analyses, where numerous simulations are required. Statistical emulators, as surrogates of these simulators, can provide predictions of the physical process in a much faster and computationally inexpensive way. They can form a prominent solution to explore thousands of scenarios that would be otherwise numerically expensive and difficult to achieve. In this work, we build a statistical emulator of the deterministic codes used to simulate submarine sliding and tsunami generation at the Rockall Bank, NE Atlantic Ocean, in two stages. First we calibrate, against observations of the landslide deposits, the parameters used in the landslide simulations. This calibration is performed under a Bayesian framework using Gaussian Process (GP) emulators to approximate the landslide model, and the discrepancy function between model and observations. Distributions of the calibrated input parameters are obtained as a result of the calibration. In a second step, a GP emulator is built to mimic the coupled landslide-tsunami numerical process. The emulator propagates the uncertainties in the distributions of the calibrated input parameters inferred from the first step to the outputs. As a result, a quantification of the uncertainty of the maximum free surface elevation at specified locations is obtained.


Knee disorders in carpet and floor layers and painters.
In an evaluation of the effect of kneeling work on the knees, 168 actively working carpet and floor layers and 146 house painters were examined with the aid of a questionnaire, a clinical examination, and radiography. Reported knee pain, knee accidents, and treatment regimens for the knees were more common among the carpet and floor layers than among the painters. Radiographic changes of the tibiofemoral joint were noted equally in the two occupational groups, but osteophytes of the patella were more common among the carpet and floor layers than among the painters. In a multivariate analysis, the determinants of osteophytosis of the knee were age, occupation, knee accidents, and smoking, and osteophytosis may be due to more frequent workbreaks from kneeling postures among smoking workers. This study indicates that kneeling work increases the risk of knee disorders and such radiographic changes that might be an initial sign of knee degeneration.


Genotoxic effects of three selected black toner powders and their dimethyl sulfoxide extracts in cultured human epithelial A549 lung cells in vitro.
Until now, the adverse effects of toner powders on humans have been considered to be minimal. However, several recent reports have suggested possible significant adverse health effects from toner dust inhalation. The aim of this study was to evaluate the genotoxic potential of black toner powders in vitro. For the study of DNA damage, A549 cells were exposed to toner-powder suspensions and to their DMSO extracts, and then subjected to the comet assay and to the in-vitro cytokinesis block micronucleus test (CB-MNvit). Cytotoxic effects of the toner samples were assessed by the erythrosin B assay. Furthermore, size, shape, and composition of the toner powders were investigated. None of the three toner powders or their DMSO extracts reduced cell viability; however, they did induce DNA damage and formed micronuclei at concentrations from 80 to 400 μg cm(-2) , although to a varying extent. All toner powders contain considerable amounts of the pigments carbon black and magnetite (Fe(3) O(4) ) as well as small amounts of polycyclic aromatic hydrocarbons (PAHs). The overall results of our in-vitro study suggest that the investigated toner-powder samples are not cytotoxic but genotoxic. From the results of the physical and chemical characterization, we conclude that metals and metalloids as components of magnetite, or PAHs as components of the carbon-bearing material, are responsible for the genotoxic effects. Further research is necessary to determine the relevance of these in-vitro observations for private and occupational toner powder exposure.


Seroprevalence of Hepatitis B Infection and Associated Risk Factors among Drug Users in Drop-in Centers of Isfahan, Iran.
Scientists perceive drug users (DUs) as a high-risk population for hepatitis B virus (HBV) infection. Effective strategies aiming at the reduction of HBV infection can be depicted when its epidemiological status is clearly defined. The present study provides new insight into associated risk factors of HBV infection and its seroepidemiological status among DUs attending drop-in centers (DICs). This was a cross-sectional study, which was implemented in 7 DICs of Isfahan province. The sample size included 539 participants. Demographic data and risk factors for HBV infection were obtained by a trained social worker using a self-made structured questionnaire. Venous blood sample was obtained and tested for hepatitis B surface antigen (HBsAg), hepatitis B surface antibody, and total hepatitis B core antibody (HBcAb) using enzyme-linked immunosorbent assay. Mean age of the participants was 31.76 ± 8.4 years. They were generally male, Iranian, urban, with an education level of high school or less. The prevalence of HBV infection (HBsAg and/or HBcAb) was 18% (88.490). Regression analysis showed that age, bloodletting, and drug injection, being the sexual partner of injecting DU (IDU), as well as frequency and duration of imprisonment positively correlated with HBV infection. Drug injection bloodletting, and being the sexual partner of IDU, as well as frequency and duration of imprisonment could be considered as contributing factors in HBV infection.


Screening for balance and mobility impairment in elderly individuals living in residential care facilities.
The rapid growth of the elderly population has resulted in a corresponding rise in the number of elderly individuals who experience disability during their lifetimes. The purpose of this study was to test the usefulness of four established clinical measures of balance, gait, and subjective perceptions of fear of falling as screening methods for referring community-dwelling elderly individuals living in residential care facilities for detailed physical therapy evaluation and possible intervention. The subjects were a convenience sample of 53 elderly individuals living in two residential care facilities for the elderly. Subjects were tested on each of four clinical measures of balance and mobility. Their performance on these measures was compared with a physical therapist's brief evaluation of disability and appropriateness for more detailed evaluation. The usefulness of these tools as screening methods was determined by calculating sensitivity and specificity levels using the physical therapist's evaluation as a standard. The sensitivity and specificity levels of the four clinical measures in their application as screening tests for referral to physical therapy were as follows: Berg Balance Scale, 84% and 78%; balance subscale of the Tinetti Performance-Oriented Mobility Assessment, 68% and 78%; gait speed, 80% and 89%; and Tinetti Fall Efficacy Scale, 59% and 82%. The combination of two tests, Berg Balance Scale and gait speed, yielded the highest sensitivity of 91% and the highest specificity of 70% when a subject tested positive on at least one test. These findings indicate the feasibility of developing screening methods for referring community-dwelling elderly individuals for a detailed physical therapy evaluation based on established clinical assessment measures, with a combination of tests measuring balance and gait demonstrating the most promising results.


Novel lactate dehydrogenase inhibitors with in vivo efficacy against Cryptosporidium parvum.
Cryptosporidium parvum is a highly prevalent zoonotic and anthroponotic protozoan parasite that causes a diarrheal syndrome in children and neonatal livestock, culminating in growth retardation and mortalities. Despite the high prevalence of C. parvum, there are no fully effective and safe drugs for treating infections, and there is no vaccine. We have previously reported that the bacterial-like C. parvum lactate dehydrogenase (CpLDH) enzyme is essential for survival, virulence and growth of C. parvum in vitro and in vivo. In the present study, we screened compound libraries and identified inhibitors against the enzymatic activity of recombinant CpLDH protein in vitro. We tested the inhibitors for anti-Cryptosporidium effect using in vitro infection assays of HCT-8 cells monolayers and identified compounds NSC158011 and NSC10447 that inhibited the proliferation of intracellular C. parvum in vitro, with IC50 values of 14.88 and 72.65 μM, respectively. At doses tolerable in mice, we found that both NSC158011 and NSC10447 consistently significantly reduced the shedding of C. parvum oocysts in infected immunocompromised mice's feces, and prevented intestinal villous atrophy as well as mucosal erosion due to C. parvum. Together, our findings have unveiled promising anti-Cryptosporidium drug candidates that can be explored further for the development of the much needed novel therapeutic agents against C. parvum infections.


DNA Damage-Inducible Transcript 4 Is an Innate Surveillant of Hair Follicular Stress in Vitamin D Receptor Knockout Mice and a Regulator of Wound Re-Epithelialization.
Mice and human patients with impaired vitamin D receptor (VDR) signaling have normal developmental hair growth but display aberrant post-morphogenic hair cycle progression associated with alopecia. In addition, VDR-/- mice exhibit impaired cutaneous wound healing. We undertook experiments to determine whether the stress-inducible regulator of energy homeostasis, DNA damage-inducible transcript 4 (Ddit4), is involved in these processes. By analyzing hair cycle activation in vivo, we show that VDR-/- mice at day 14 exhibit increased Ddit4 expression within follicular stress compartments. At day 29, degenerating VDR-/- follicular keratinocytes, but not bulge stem cells, continue to exhibit an increase in Ddit4 expression. At day 47, when normal follicles and epidermis are quiescent and enriched for Ddit4, VDR-/- skin lacks Ddit4 expression. In a skin wound healing assay, the re-epithelialized epidermis in wildtype (WT) but not VDR-/- animals harbor a population of Ddit4- and Krt10-positive cells. Our study suggests that VDR regulates Ddit4 expression during epidermal homeostasis and the wound healing process, while elevated Ddit4 represents an early growth-arresting stress response within VDR-/- follicles.


Pictures of appetizing foods activate gustatory cortices for taste and reward.
Increasing research indicates that concepts are represented as distributed circuits of property information across the brain's modality-specific areas. The current study examines the distributed representation of an important but under-explored category, foods. Participants viewed pictures of appetizing foods (along with pictures of locations for comparison) during event-related fMRI. Compared to location pictures, food pictures activated the right insula/operculum and the left orbitofrontal cortex, both gustatory processing areas. Food pictures also activated regions of visual cortex that represent object shape. Together these areas contribute to a distributed neural circuit that represents food knowledge. Not only does this circuit become active during the tasting of actual foods, it also becomes active while viewing food pictures. Via the process of pattern completion, food pictures activate gustatory regions of the circuit to produce conceptual inferences about taste. Consistent with theories that ground knowledge in the modalities, these inferences arise as reenactments of modality-specific processing.


Behavioral and biochemical evidence of opioidergic involvement in cocaine sensitization.
Chronic administration of cocaine produces sensitization to its behavioral effects in humans and experimental animals. In the present study, rats treated with cocaine (10 mg/kg, i.p.) once daily for 10 days showed an enhancement in the acute drug stimulation of locomotor activity and stereotypy. Biochemical analysis in the nucleus accumbens of chronic cocaine-treated animals indicated that sensitization of D1 dopamine (DA) receptors had also developed. In fact, stimulation of adenylyl cyclase activity by DA was increased in nucleus accumbens membranes from sensitized rats. Our findings suggest that a novel postsynaptic mechanism, i.e., an increased DA-D1 receptor function, may play a role in the sensitization. A causal relationship between the two events is supported by the observation that neither motor behavioral sensitization nor DA-dependent adenylyl cyclase hyperactivity developed when the opiate antagonist naltrexone (2 mg/kg, s.c.) was given daily for 10 days before cocaine. When given alone, naltrexone was inactive in all respects, which rules out any unspecific action and suggests that its effects may be due to competition at receptors with endogenous opioids mobilized by cocaine. This was indirectly supported by the finding that desensitization of opioid inhibition of adenylyl cyclase developed in nucleus accumbens membranes of cocaine-sensitized rats. Chronic blockade of opioid receptors by naltrexone also counteracted the reinforcing properties of cocaine; conditioned place preference, clearly displayed by cocaine-treated animals, was antagonized in a dose-related manner. Overall, these results confirm that endogenous opioid peptides play an important role in cocaine addiction.


Antagonism of CD95 signaling blocks butyrate induction of apoptosis in young adult mouse colonic cells.
There is great interest in utilizing butyrate as a chemopreventive agent for colon tumorigenesis because of its ability to promote apoptosis in colon tumor cell lines. Because CD95 (APO-1/Fas) transduces signals resulting in apoptosis, we tested the hypothesis that butyrate-dependent colonocyte apoptosis is mediated by this death receptor. Butyrate (1 mM) exposure for 24 h upregulated expression of Fas and its ligand in young adult mouse colon (YAMC) cells. To delineate the proapoptotic effect of butyrate and to avoid the confounding effects of detachment from the extracellular matrix, adherent cell apoptosis was monitored as loss of plasma membrane asymmetry and dissipation of mitochondrial membrane potential (DeltaPsi(mt)) by laser cytometry. Soluble Fas receptor protein (Fas:Fc chimera) and caspase inhibitors (z-VAD-fmk and z-IETD-fmk) blocked butyrate induction of apoptosis. Treatment with Fas agonistic antibody (clone Jo-2) significantly induced cell death, indicating that Fas in colonocytes is functional. In addition, butyrate promoted apoptosis by inducing loss of DeltaPsi(mt) and phospholipid asymmetry of the plasma membrane after 12 and 24 h of exposure, respectively, before cell detachment. Therefore, Fas receptor-dependent signal transduction is involved in butyrate induction of apoptosis in colonocytes.