Shared decision-making in tinnitus care - An exploration of clinical encounters.
This study examined clinical encounters between clinicians and patients to determine current practice for the diagnosis and treatment of tinnitus. The objective was to develop an understanding of the ideal clinical encounter that would facilitate genuine shared decision-making. Video ethnography was used to examine clinical encounters for the diagnosis and treatment of tinnitus. Clinical encounters were video-recorded. Patients were interviewed individually following their clinic appointment. Data were analysed using constant comparison techniques from Grounded Theory. Initial inductive analyses were then considered against theoretical conceptualizations of the clinician-patient relationship and of the clinical encounter. Alignment between clinician and patient was found to be essential to a collaborative consultation and to shared decision-making. Clinician groups demonstrated variation in behaviour in the encounter; some asked closed questions and directed the majority of the consultation; others asked open questions and allowed patients to lead the consultation. A shift away from aetiology and physiological tests is needed so that tinnitus is managed as a persistent unexplained set of symptoms. This uncertainty is challenging for the medical professionals; lessons could be learned from the use of therapeutic skills. Further research is required to test techniques, such as the use of decision aids, to determine how we might create the ideal clinical encounter. Statement of contribution What is already known on this subject? Tinnitus is a condition in which sound is heard in the absence of an external source. Current approaches to managing tinnitus vary depending on clinical site (Hoare & Hall, ). In most instances, tinnitus does not have a straightforward medical cause. Tinnitus care is challenging to traditional biomedical encounters because the process of diagnosis may not lead to a defined treatment. Clinicians are required to consider not only what the tinnitus sounds like but more importantly, what it means for the affected individual. This requires a careful and skilled approach to eliciting a patient's current behaviour, coping, and preferences for both outcomes and treatment approaches. What does this study add? We provide the first in-depth description of decision-making in clinical services for tinnitus. Findings suggest a shift in focus is required to move away from the current prioritization of the biomedical treatment of tinnitus. There is variation to the extent different clinicians were able to deal with the uncertainty presented by the symptoms of tinnitus.


Energy restriction lowers the expression of genes linked to inflammation, the cytoskeleton, the extracellular matrix, and angiogenesis in mouse adipose tissue.
Using high-density oligonucleotide microarrays, we examined the actions of energy restriction (ER) on the expression of >11,000 genes in epididymal white adipose tissue (WAT) of 10- to 11-mo-old male C57Bl6 mice. Four groups were studied: controls not subjected to food restriction (CO), food-restricted 18 h before being killed (FR), short-term ER for 23 d (SER), and long-term ER for 9 mo (LER). As we reported previously, compared with CO mice, FR and SER minimally influenced the gene expression profiles; however, 345 transcripts of 6,266 genes determined to be expressed in WAT were significantly altered by LER. We focus here on the 109 (31%) of these genes that were involved in either inflammation (56 genes), cytoskeleton (16 genes), extracellular matrix (23 genes), or angiogenesis (14 genes). Among these 109 genes, 104 transcripts (95%) were down regulated by LER. Western blotting for heat shock protein 47 and osteonectin, and immunohistochemical staining for hypoxia inducible factor (HIF)-1alpha), supported the microarray data that LER down regulated the expressions of these genes. Additionally, a 75% reduction in adipocyte size with LER reflected the change in the expression of genes involved in cell morphology. Our findings provide evidence that LER suppresses the expression of genes encoding inflammatory molecules in WAT while promoting structural remodeling of the cytoskeleton, extracellular matrix, and vasculature. These alterations may play an important role in the protection against WAT-derived inflammation and in lifespan extension by LER.


Paracrine inhibition of prion propagation by anti-PrP single-chain Fv miniantibodies.
Prion diseases are characterized by the deposition of PrP(Sc), an abnormal form of the cellular prion protein PrP(C). A growing body of evidence suggests that antibodies to PrP(C) can antagonize deposition of PrP(Sc). However, host tolerance hampers the induction of immune responses to PrP(C), and cross-linking of PrP(C) by bivalent anti-PrP antibodies is neurotoxic. In order to obviate these problems, we explored the antiprion potential of recombinant single-chain antibody (scFv) fragments. scFv fragments derived from monoclonal anti-PrP antibody 6H4, flagged with c-myc and His6 tags, were correctly processed and secreted by mammalian RD-4 rhabdomyosarcoma cells. When cocultured with cells secreting anti-PrP scFv, chronically prion-infected neuroblastoma cells ceased to produce PrP(Sc), even if antibody-producing cells were physically separated from target cells in transwell cultures. Expression of scFv with irrelevant specificity, or of similarly tagged molecules, was not curative. Therefore, eukaryotically expressed scFv exerts a paracrine antiprion activity. The effector functions encoded by immunoglobulin constant domains are unnecessary for this effect. Because of their small size and their monovalent binding, scFv fragments may represent candidates for gene transfer-based immunotherapy of prion diseases.


The potential value of biomarkers in diagnosis and staging of COPD and exacerbations.
There is an unmet need in the diagnosis, phenotyping, and staging of COPD that could potentially be fulfilled by a validated molecular biomarker. Many promising candidates have been investigated, and some have been shown to be useful in certain situations. However, to date there is no outstanding disease-specific biomarker for widespread clinical application for patients in the stable state. Given the functional, social, and financial importance of exacerbations of COPD, it would be very useful to be able to employ a biomarker to aid optimal treatment and predict clinical outcome from the acute episode. Although serum C-reactive protein (CRP) is not specific to COPD, its use as a molecular biomarker in the stable state and at exacerbation has been studied extensively, and it remains the most commonly measured molecular biomarker in routine secondary care practice. Utilizing biomarkers in combinations may ultimately prove more useful. Airway-derived biomarkers and their relationships with outcome measures require further longitudinal study as well as refinement of sampling techniques to make them more broadly applicable. There is substantial ongoing investigation of many biomarkers that we are hopeful will advance the field for the benefit of our patients.


Breast-feeding and atopic disease: a cohort study from childhood to middle age.
The literature regarding the association between breast-feeding and atopic diseases has been contradictory. We have assessed the relationship between breast-feeding and atopic disorders in a cohort followed into middle age. The Tasmanian Asthma Study is a population-based prospective cohort study that has followed participants from the age of 7 to 44 years. Exclusive breast-feeding in the first 3 months of life was examined as a risk factor for atopic diseases by using multiple logistic regression and generalized estimating equation analyses. At age 7 years, exclusively breast-fed children with a maternal history of atopy had a marginally lesser risk of current asthma than those not exclusively breast-fed (odds ratio [OR], 0.8; 95% CI, 0.6-1.0). However, after age 7 years, the risk reversed, and exclusively breast-fed children had an increased risk of current asthma at 14 (OR, 1.46; 95% CI, 1.02-2.07), 32 (OR, 1.84; 95% CI, 1.06-3.3), and 44 (OR, 1.57; 95% CI, 1.15-2.14) years. Exclusively breast-fed children also had a reduced risk of food allergy at age 7 years but an increased risk of food allergy (OR, 1.26; 95% CI, 1.1-1.5) and allergic rhinitis (OR, 1.2; 95% CI, 1.0-1.3) at 44 years. Exclusively breast-fed babies with a maternal history of atopy were less likely to develop asthma before the age of 7 years, but more likely to develop asthma after the age of 7 years. The current recommendation to breast-feed high-risk infants for protection against early wheezing illness can be confirmed. However, the recommendation should be reconsidered for protection against allergic asthma and atopy in the longer term.


The resumption of consumption -- a review on tuberculosis.
Among all infectious diseases that afflict humans, tuberculosis (TB) remains the deadliest. At present, epidemiologists estimate that one-third of the world population is infected with tubercle bacilli, which is responsible for 8 to 10 million new cases of TB and 3 million deaths annually throughout the world. Approximately 95% of new cases and 98% of deaths occur in developing nations, generally due to the few resources available to ensure proper treatment and where human immunodeficiency virus (HIV) infections are common. In 1882, Dr Robert Koch identified an acid-fast bacterium, Mycobacterium tuberculosis, as the causative agent of TB. Thirty-nine years later, BCG vaccine was introduced for human use, and became the most widely used prophylactic strategy to fight TB in the world. The discovery of the properties of first-line antimycobacterial drugs in the past century yielded effective chemotherapies, which considerably decreased TB mortality rates worldwide. The later introduction of some additional drugs to the arsenal used to treat TB seemed to provide an adequate number of effective antimicrobial agents. The modern, standard short-course therapy for TB recommended by the World Health Organization is based on a four-drug regimen that must be strictly followed to prevent drug resistance acquisition, and relies on direct observation of patient compliance to ensure effective treatment. Mycobacteria show a high degree of intrinsic resistance to most antibiotics and chemotherapeutic agents due to the low permeability of its cell wall. Nevertheless, the cell wall barrier alone cannot produce significant levels of drug resistance. M. tuberculosis mutants resistant to any single drug are naturally present in any large bacterial population, irrespective of exposure to drugs. The frequency of mutants resistant to rifampicin and isoniazid, the two principal antimycobacterial drugs currently in use, is relatively high and, therefore, the large extra-cellular population of actively metabolizing and rapidly growing tubercle bacilli in cavitary lesions will contain organisms which are resistant to a single drug. Consequently, monotherapy or improperly administered two-drug therapies will select for drug-resistant mutants that may lead to drug resistance in the entire bacterial population. Thereby, despite the availability of effective chemotherapy and the moderately protective vaccine, new anti-TB agents are urgently needed to decrease the global incidence of TB. The resumption of TB, mainly caused by the emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains and HIV epidemics, led to an increased need to understand the molecular mechanisms of drug action and drug resistance, which should provide significant insight into the development of newer compounds. The latter should be effective to combat both drug-susceptible and MDR/XDR-TB.


Variation in the maternal corticotrophin releasing hormone-binding protein (CRH-BP) gene and birth weight in Blacks, Hispanics and Whites.
Given the unique role of the corticotrophin-releasing hormone (CRH) system in human fetal development, the aim of our study was to estimate the association of birth weight with DNA sequence variation in three maternal genes involved in regulating CRH production, bioavailability and action: CRH, CRH-Binding Protein (CRH-BP), and CRH type 1 receptor (CRH-R1), respectively, in three racial groups (African-Americans, Hispanics, and non-Hispanic Whites). Our study was carried out on a population-based sample of 575 mother-child dyads. We resequenced the three genes in mouse-human hybrid somatic cell lines and selected SNPs for genotyping. A significant association was observed in each race between birth weight and maternal CRH-BP SNP genotypes. Estimates of linkage disequilibrium and haplotypes established three common haplotypes marked by the rs1053989 SNP in all three races. This SNP predicted significant birth weight variation after adjustment for gestational age, maternal BMI, parity, and smoking. African American and Hispanic mothers carrying the A allele had infants whose birth weight was on average 254 and 302 grams, respectively, less than infants having C/C mothers. Non-Hispanic White mothers homozygous for the A allele had infants who were on average 148 grams less than those infants having A/C and C/C mothers. The magnitudes of the estimates of the birth weight effects are comparable to the combined effects of multiple SNPs reported in a recent meta-analysis of 6 GWAS studies and is quantitatively larger than that associated with maternal cigarette smoking. This effect was persistent across subpopulations that vary with respect to ancestry and environment.


Modes of Transmission of Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) and Factors Influencing on the Airborne Transmission: A Review.
The multiple modes of SARS-CoV-2 transmission including airborne, droplet, contact, and fecal-oral transmissions that cause coronavirus disease 2019 (COVID-19) contribute to a public threat to the lives of people worldwide. Herein, different databases are reviewed to evaluate modes of transmission of SARS-CoV-2 and study the effects of negative pressure ventilation, air conditioning system, and related protection approaches of this virus. Droplet transmission was commonly reported to occur in particles with diameter >5 µm that can quickly settle gravitationally on surfaces (1-2 m). Instead, fine and ultrafine particles (airborne transmission) can stay suspended for an extended period of time (≥2 h) and be transported further, e.g., up to 8 m through simple diffusion and convection mechanisms. Droplet and airborne transmission of SARS-CoV-2 can be limited indoors with adequate ventilation of rooms, by routine disinfection of toilets, using negative pressure rooms, using face masks, and maintaining social distancing. Other preventive measures recommended include increasing the number of screening tests of suspected carriers of SARS-CoV-2, reducing the number of persons in a room to minimize sharing indoor air, and monitoring people's temperature before accessing a building. The work reviews a body of literature supporting the transmission of SARS-CoV-2 through air, causing COVID-19 disease, which requires coordinated worldwide strategies.


Influence Function Learning in Information Diffusion Networks.
Can we learn the influence of a set of people in a social network from cascades of information diffusion? This question is often addressed by a two-stage approach: first learn a diffusion model, and then calculate the influence based on the learned model. Thus, the success of this approach relies heavily on the correctness of the diffusion model which is hard to verify for real world data. In this paper, we exploit the insight that the influence functions in many diffusion models are coverage functions, and propose a novel parameterization of such functions using a convex combination of random basis functions. Moreover, we propose an efficient maximum likelihood based algorithm to learn such functions directly from cascade data, and hence bypass the need to specify a particular diffusion model in advance. We provide both theoretical and empirical analysis for our approach, showing that the proposed approach can provably learn the influence function with low sample complexity, be robust to the unknown diffusion models, and significantly outperform existing approaches in both synthetic and real world data.


Wavelength tuning in optically mode-locked semiconductor fiber ring lasers with linearly chirped fiber Bragg gratings.
We demonstrate wavelength tuning in single-wavelength and multiwavelength semiconductor fiber ring lasers that are mode locked with an optically injected control signal. A semiconductor optical amplifier is used to provide gain as well as to function as an optically controlled mode-locking element. Linearly chirped fiber Bragg gratings--single or superimposed--are used to define the lasing wavelengths as well as to provide wavelength tunability and allow for multiwavelength operation. We obtain pulses of tens of picoseconds in duration when we inject a sinusoidal optical control signal into the laser cavity, and we can tune the lasing wavelength(s) over the reflection bandwidth(s) of the grating(s) by simply changing the frequency of the injected control signal.


Neutrophil extracellular traps (NETs) as markers of disease severity in COVID-19.
In severe cases of coronavirus disease 2019 (COVID-19), viral pneumonia progresses to respiratory failure. Neutrophil extracellular traps (NETs) are extracellular webs of chromatin, microbicidal proteins, and oxidant enzymes that are released by neutrophils to contain infections. However, when not properly regulated, NETs have potential to propagate inflammation and microvascular thrombosis, including in the lungs of patients with acute respiratory distress syndrome. While elevated levels of blood neutrophils predict worse outcomes in COVID-19, the role of NETs has not been investigated. We now report that sera from patients with COVID-19 (n=50 patients, n=84 samples) have elevated levels of cell-free DNA, myeloperoxidase(MPO)-DNA, and citrullinated histone H3 (Cit-H3); the latter two are highly specific markers of NETs. Highlighting the potential clinical relevance of these findings, cell-free DNA strongly correlated with acute phase reactants including C-reactive protein, D-dimer, and lactate dehydrogenase, as well as absolute neutrophil count. MPO-DNA associated with both cell-free DNA and absolute neutrophil count, while Cit-H3 correlated with platelet levels. Importantly, both cell-free DNA and MPO-DNA were higher in hospitalized patients receiving mechanical ventilation as compared with hospitalized patients breathing room air. Finally, sera from individuals with COVID-19 triggered NET release from control neutrophils in vitro. In summary, these data reveal high levels of NETs in many patients with COVID-19, where they may contribute to cytokine release and respiratory failure. Future studies should investigate the predictive power of circulating NETs in longitudinal cohorts, and determine the extent to which NETs may be novel therapeutic targets in severe COVID-19.


Glycine restores the anabolic response to leucine in a mouse model of acute inflammation.
Amino acids, especially leucine, potently stimulate protein synthesis and reduce protein breakdown in healthy skeletal muscle and as a result have received considerable attention as potential treatments for muscle wasting. However, the normal anabolic response to amino acids is impaired during muscle-wasting conditions. Although the exact mechanisms of this anabolic resistance are unclear, inflammation and ROS are believed to play a central role. The nonessential amino acid glycine has anti-inflammatory and antioxidant properties and preserves muscle mass in calorie-restricted and tumor-bearing mice. We hypothesized that glycine would restore the normal muscle anabolic response to amino acids under inflammatory conditions. Relative rates of basal and leucine-stimulated protein synthesis were measured using SUnSET methodology 4 h after an injection of 1 mg/kg lipopolysaccharide (LPS). Whereas leucine failed to stimulate muscle protein synthesis in LPS-treated mice pretreated with l-alanine (isonitrogenous control), leucine robustly stimulated protein synthesis (+51%) in mice pretreated with 1 g/kg glycine. The improvement in leucine-stimulated protein synthesis was accompanied by a higher phosphorylation status of mTOR, S6, and 4E-BP1 compared with l-alanine-treated controls. Despite its known anti-inflammatory action in inflammatory cells, glycine did not alter the skeletal muscle inflammatory response to LPS in vivo or in vitro but markedly reduced DHE staining intensity, a marker of oxidative stress, in muscle cross-sections and attenuated LPS-induced wasting in C2C12 myotubes. Our observations in male C57BL/6 mice suggest that glycine may represent a promising nutritional intervention for the attenuation of skeletal muscle wasting.


Evidence for addiction-like behavior in the rat.
Although the voluntary intake of drugs of abuse is a behavior largely preserved throughout phylogeny, it is currently unclear whether pathological drug use ("addiction") can be observed in species other than humans. Here, we report that behaviors that resemble three of the essential diagnostic criteria for addiction appear over time in rats trained to self-administer cocaine. As in humans, this addiction-like behavior is present only in a small proportion of subjects using cocaine and is highly predictive of relapse after withdrawal. These findings provide a new basis for developing a true understanding and treatment of addiction.


Does detection of chlamydial antibodies by microimmunofluorescence help in managing chlamydial lower genital tract infection in women?
A total of 113 women thought to have chlamydial infection of the lower genital tract were studied prospectively to evaluate the effect of antibiotic treatment on antibodies to chlamydiae detected by microimmunofluorescence. Of them, 81 were randomly selected for treatment with a two week course of either triple tetracycline or erythromycin stearate, and 32 who had microimmunofluorescent antibodies to, but did not yield cultures for, chlamydiae were used as controls and left untreated. Results for the treated patients showed that 22 (27%) had at least a fourfold fall in the microimmunofluorescent titre, but there was a similar rise in titre in 14 (17%), and the titre remained unaltered in 45 (56%) patients. In the control group 10 (31%) patients had at least a fourfold fall in titre, but there was a similar rise in titre in seven (22%), and it remained unaltered in 15 (47%) patients. The differences between these percentages in treated and untreated patients were not significant.


Synchronization of world economic activity.
Common dynamical properties of business cycle fluctuations are studied in a sample of more than 100 countries that represent economic regions from all around the world. We apply the methodology of multivariate singular spectrum analysis (M-SSA) to identify oscillatory modes and to detect whether these modes are shared by clusters of phase- and frequency-locked oscillators. An extension of the M-SSA approach is introduced to help analyze structural changes in the cluster configuration of synchronization. With this novel technique, we are able to identify a common mode of business cycle activity across our sample, and thus point to the existence of a world business cycle. Superimposed on this mode, we further identify several major events that have markedly influenced the landscape of world economic activity in the postwar era.


Towards Improving Skin Cancer Diagnosis by Integrating Microarray and RNA-Seq Datasets.
Many clinical studies have revealed the high biological similarities existing among different skin pathological states. These similarities create difficulties in the efficient diagnosis of skin cancer, and encourage to study and design new intelligent clinical decision support systems. In this sense, gene expression analysis can help find differentially expressed genes (DEGs) simultaneously discerning multiple skin pathological states in a single test. The integration of multiple heterogeneous transcriptomic datasets requires different pipeline stages to be properly designed: from suitable batch merging and efficient biomarker selection to automated classification assessment. This article presents a novel approach addressing all these technical issues, with the intention of providing new sights about skin cancer diagnosis. Although new future efforts will have to be made in the search for better biomarkers recognizing specific skin pathological states, our study found a panel of 8 highly relevant multiclass DEGs for discerning up to 10 skin pathological states: 2 healthy skin conditions a priori, 2 cataloged precancerous skin diseases and 6 cancerous skin states. Their power of diagnosis over new samples was widely tested by previously well-trained classification models. Robust performance metrics such as overall and mean multiclass F1-score outperformed recognition rates of 94% and 80%, respectively. Clinicians should give special attention to highlighted multiclass DEGs that have high gene expression changes present among them, and understand their biological relationship to different skin pathological states.


Treatment of migraine with pulsing electromagnetic fields: a double-blind, placebo-controlled study.
The effect of exposure to pulsing electromagnetic fields on migraine activity was evaluated by having 42 subjects (34 women and 8 men), who met the International Headache Society's criteria for migraine, participate in a double-blind, placebo-controlled study. Each subject kept a 1-month, pretreatment, baseline log of headache activity prior to being randomized to having either actual or placebo pulsing electromagnetic fields applied to their inner thighs for 1 hour per day, 5 days per week, for 2 weeks. After exposure, all subjects kept the log for at least 1 follow-up month. During the first month of follow-up, 73% of those receiving actual exposure reported decreased headaches (45% good decrease, 14% excellent decrease) compared to half of those receiving the placebo (15% worse, 20% good, 0% excellent). Ten of the 22 subjects who had actual exposure received 2 additional weeks of actual exposure after their initial 1-month follow-up. All showed decreased headache activity (50% good, 38% excellent). Thirteen subjects from the actual exposure group elected not to receive additional exposure. Twelve of them showed decreased headache activity by the second month (29% good, 43% excellent). Eight of the subjects in the placebo group elected to receive 2 weeks of actual exposure after the initial 1-month follow-up with 75% showing decreased headache activity (38% good, 38% excellent). In conclusion, exposure of the inner thighs to pulsing electromagnetic fields for at least 3 weeks is an effective, short-term intervention for migraine, but not tension headaches.


The Impact of High-Flow Nasal Cannula Use on Patient Mortality and the Availability of Mechanical Ventilators in COVID-19.
Rationale: How to provide advanced respiratory support for coronavirus disease (COVID-19) to maximize population-level survival while optimizing mechanical ventilator access is unknown.Objectives: To evaluate the use of high-flow nasal cannula for COVID-19 on population-level mortality and ventilator availability.Methods: We constructed dynamical (deterministic) simulation models of high-flow nasal cannula and mechanical ventilation use for COVID-19 in the United States. Model parameters were estimated through consensus based on published literature, local data, and experience. We had the following two outcomes: 1) cumulative number of deaths and 2) days without any available ventilators. We assessed the impact of various policies for the use of high-flow nasal cannula (with or without "early intubation") versus a scenario in which high-flow nasal cannula was unavailable.Results: The policy associated with the fewest deaths and the least time without available ventilators combined the use of high-flow nasal cannula for patients not urgently needing ventilators with the use of early mechanical ventilation for these patients when at least 10% of ventilator supply was not in use. At the national level, this strategy resulted in 10,000-40,000 fewer deaths than if high-flow nasal cannula were not available. In addition, with moderate national ventilator capacity (30,000-45,000 ventilators), this strategy led to up to 25 (11.8%) fewer days without available ventilators. For a 250-bed hospital with 100 mechanical ventilators, the availability of 13, 20, or 33 high-flow nasal cannulas prevented 81, 102, and 130 deaths, respectively.Conclusions: The use of high-flow nasal cannula coupled with early mechanical ventilation when supply is sufficient results in fewer deaths and greater ventilator availability.


High precision Neisseria gonorrhoeae variant and antimicrobial resistance calling from metagenomic Nanopore sequencing.
The rise of antimicrobial-resistant Neisseria gonorrhoeae is a significant public health concern. Against this background, rapid culture-independent diagnostics may allow targeted treatment and prevent onward transmission. We have previously shown metagenomic sequencing of urine samples from men with urethral gonorrhea can recover near-complete N. gonorrhoeae genomes. However, disentangling the N. gonorrhoeae genome from metagenomic samples and robustly identifying antimicrobial resistance determinants from error-prone Nanopore sequencing is a substantial bioinformatics challenge. Here, we show an N. gonorrhoeae diagnostic workflow for analysis of metagenomic sequencing data obtained from clinical samples using R9.4.1 Nanopore sequencing. We compared results from simulated and clinical infections with data from known reference strains and Illumina sequencing of isolates cultured from the same patients. We evaluated three Nanopore variant callers and developed a random forest classifier to filter called SNPs. Clair was the most suitable variant caller after SNP filtering. A minimum depth of 20× reads was required to confidently identify resistant determinants over the entire genome. Our findings show that metagenomic Nanopore sequencing can provide reliable diagnostic information in N. gonorrhoeae infection.


Interaction between pain, movement, and physical activity: short-term benefits, long-term consequences, and targets for treatment.
Movement is changed in pain. This presents across a spectrum from subtle changes in the manner in which a task is completed to complete avoidance of a function and could be both a cause and effect of pain/nociceptive input and/or injury. Movement, in a variety of forms, is also recommended as a component of treatment to aid the recovery in many pain syndromes. Some argue it may not be sufficient to simply increase activity, whereas others defend a necessity to consider how a person moves. There is unlikely to be a simple relationship between pain and movement, as both too little and too much movement could be suboptimal for the health of the tissues. The interaction between pain, (re)injury, and movement is surprisingly unclear. Traditional theories to explain adaptation in the motor system in pain are unable to account for the variability observed in laboratory and clinical practice. New theories are required. Treatments that focus on physical activity and exercise are the cornerstone of management of many pain conditions, but the effect sizes are modest. There is limited consensus when, if, and how interventions may be individualized and combined. The aim of this narrative review was to present current understanding of the interaction between movement and pain; as a cause or effect of pain, and in terms of the role of movement (physical activity and exercise) in recovery of pain and restoration of function.


Three-Catheter Technique for Ablation of Left-Sided Accessory Pathways in Wolff-Parkinson-White is Less Expensive and Equally Successful When Compared to a Five-Catheter Technique.
To compare the efficacy, safety, and cost-effectiveness of a three-catheter approach with a conventional five-catheter approach for the mapping and ablation of supraventricular tachycardia in pediatric patients with Wolff-Parkinson-White Syndrome (WPW) and concealed accessory pathways (APs). A retrospective review from 2008 to 2012 of patients less than 21 years with WPW who underwent a three-catheter radiofrequency (RF) ablation of a left-sided AP (ablation, right ventricular [RV] apical, and coronary sinus [CS] decapolar catheters) was performed. The three-catheter group was compared to a control group who underwent a standard five-catheter (ablation, RV apical, CS decapolar, His catheter, and right atrial catheter) ablation for the treatment of left-sided WPW or concealed AP. Demographics, ablation outcomes, and costs were compared between groups. Twenty-eight patients met inclusion criteria with 28 control patients. The groups did not differ in gender, age, weight, or body surface area. Locations of the AP on the mitral annulus were similar between the groups. All patients were ablated via transseptal approach. Note that 28 of 28 in the three-catheter group (100%) and 27 of 28 (96%) controls were acutely successfully ablated (P = 0.31). No complications were encountered. There was no difference in procedural time, time to loss of AP conduction, or number of RF applications. Use of the three-catheter technique resulted in a total savings of $2,465/case, which includes the $680 savings from using fewer catheters as well as the savings from a shortened procedure time. Ablation in patients with WPW and a left-sided AP can be performed using three catheters with similar efficacy and safety while offering significant cost savings compared to a conventional five-catheter approach.


Intramolecular Interactions of Highly π-Conjugated Perylenediimide Oligomers Probed by Single-Molecule Spectroscopy.
Highly π-conjugated perylenediimide (PDI) oligomers are promising low band gap organic materials for various applications in optoelectronics. In this work, individual fluorescence dynamics of ethynylene- and butadiynylene-bridged dimeric and trimeric PDIs (PEP, PBP, and PEPEP) were monitored and analyzed by single-molecule fluorescence spectroscopy to gain information on the degree of extension of π-conjugation through the acetylene bridge in PDI multichromophores. The simultaneous measurements of fluorescence intensity, lifetime, and spectrum indicate a sequential decrease in π-conjugation upon photobleaching of PDI monomer units. Furthermore, Huang-Rhys (HR) factors, S, are obtained to evaluate the degree of electronic coupling in view of π-conjugation and overall rigidity between the PDI units in PDI oligomers at the single-molecule level. In addition, butadiynylene-bridged dimeric PDI (PBP) reveals conformational heterogeneity due to the long butadiynylene linker. These results suggest a new way to control the photophysical properties of the PDI multichromophoric system by expansion of π-conjugation and modification with different linker groups.


Exploitation of host cell cytoskeleton and signalling during Listeria monocytogenes entry into mammalian cells.
Deciphering how Listeria monocytogenes exploits the host cell machinery to invade mammalian cells during infection isa key issue for the understanding how this food-borne pathogen causes a pleiotropic disease ranging from gastro-enteritis to meningitis and abortions. Using multidisciplinary approaches, essentially combining bacterial genetics and cell biology, we have identified two bacterial proteins critical for entry into target cells, InlA and InlB. Their cellular ligands have been also identified: InlA interacts with the adhesion molecule E-cadherin, while InlB interacts with the receptor for the globular head of the complement factor Clq (gClq-R), with the hepatocyte growth factor receptor (c-Met) and with glycosaminoglycans(including heparan sulphate). The dynamic interaction between these cellular receptors and the actin cytoskeleton is currently under investigation. Several intracellular molecules have been recognized as key effectors for Listeria entry into target cells,including catenins (implicated in the connection of E-cadherin to actin) and the actin depolymerising factor/cofilin (involved in the rearrangement of the cytoskeleton in the InlB-dependent internalisation pathway). At the organism level, species specificity has been discovered concerning the interaction between InlA and E-cadherin, leading to the generation of transgenic mice expressing the human E-cadherin, in which the critical role of InlA in the crossing of the intestinal barrier has been clearly determined. Listeria appears as an instrumental model for addressing critical questions concerning both the complex process of bacterial pathogenesis and also fundamental molecular processes, such as phagocytosis.


Mechanism of the chromosomal translocation t(14;18) in lymphoma: detection of a 45-Kd breakpoint binding protein.
The translocation t(14;18) between the BCL-2 oncogene and the Ig heavy chain (IgH) gene provides the molecular basis for the development of follicular lymphomas. The illegitimate recombination occurs in early B cells. While V(D)J-recombinase is most likely involved on the chromosome 14 part, little is known about the mechanism of breakage on chromosome 18. We investigated the BCL-2 breakpoint regions for their structural vulnerability and protein binding capacity. We found that the major breakpoint region (mbr) contains an S1 nuclease-sensitive site and is the target of an endogenous nuclease present in early B cells. A 45 Kd nuclear protein (bp45) from early B cell extracts binds to a homopurine-homopyrimidine stretch (GGGAGGACGGGAGGAAGGCG) in the mbr, which is homologous to a recombinatorial element in Escherichia coli (CHI). The protein also binds to homologous sequences in the minor breakpoint cluster region (mcr) and in the IgH locus. The localization of the binding sites on both chromosomes as well as the tissue distribution of bp45 suggest that this protein-DNA interaction is involved in the translocation t(14;18). The DNA binding motif is also present at other translocation breakpoints indicating a more general role for this mechanism.


Development of a fully automated toxicological LC-MS(n) screening system in urine using online extraction with turbulent flow chromatography.
In clinical toxicology, fast and specific methods are necessary for the screening of different classes of drugs. Therefore, an online extraction high-performance liquid chromatography coupled to mass spectrometry (LC-MS(n)) screening method using a MS(2) and MS(3) spectral library for the identification of xenobiotic substances has been developed and validated. Samples were run twice, once native and once after enzymatic hydrolysis. Internal standards and buffer were added to the urine samples. Following centrifugation, the supernatant was injected into the system. Extraction was performed by online turbulent flow chromatography. The chromatographic separation was achieved using a Phenyl/Hexyl column. For detection, a linear ion trap, equipped with an APCI interface, was used and the different compounds were identified using a MS(2) and MS(3) spectral library containing 356 compounds. The turnaround time to report the results of the screening including hydrolysis was approximately 2 h. About 92% of the 356 substances could be identified with a limit of identification below 100 ng/ml. The recovery and matrix effect experiments showed suitable results, and in six drug-free urine samples of healthy volunteers analyzed for selectivity, no substances have been identified. Carryover could be well controlled, and the method had a good reproducibility. The comparison of the results of 103 real patient urine samples showed a good agreement between the existing GC-MS and LC-MS methods with offline extraction and the new online extraction LC-MS(n) screening method. The presented method allows a fast and sensitive analysis of a broad range of compounds.


Complete Genome Sequence of Elephant Endotheliotropic Herpesvirus 4, the First Example of a GC-Rich Branch Proboscivirus.
A novel group of mammalian DNA viruses called elephant endotheliotropic herpesviruses (EEHVs) belonging to the Proboscivirus genus has been associated with nearly 100 cases of highly lethal acute hemorrhagic disease in young Asian elephants worldwide. The complete 180-kb genomes of prototype strains from three AT-rich branch viruses, EEHV1A, EEHV1B, and EEHV5, have been published. However, less than 6 kb of DNA sequence each from EEHV3, EEHV4, and EEHV7 showed them to be a hugely diverged second major branch with GC-rich characteristics. Here, we determined the complete 206-kb genome of EEHV4(Baylor) directly from trunk wash DNA by next-generation sequencing and de novo assembly procedures. Among a total of 119 genes with an overall colinear organization similar to those of the AT-rich EEHVs, major features of EEHV4 include a family of 26 paralogous 7xTM and vGPCR-like genes plus 25 novel or missing genes. The genome also contains an unusual distribution of tracts of 5 to 11 successive A or T nucleotides in intergenic domains between the mostly much higher GC content protein coding regions. Furthermore, an extremely high GC-rich bias in the third wobble position of codons clearly delineates the coding regions for many but not all proteins. There are also two novel captured cellular genes, including a C-type lectin (vECTL) and an O-linked acetylglucosamine transferase (vOGT), as well as an unusually large and complex Ori-Lyt dyad symmetry domain. Finally, 30 kb from a second strain proved to include three small chimeric domains, indicating the existence of distinct EEHV4A and EEHV4B subtypes. IMPORTANCE Multiple species of herpesviruses from three different lineages of the Proboscivirus genus (EEHV1/6, EEHV2/5, and EEHV3/4/7) infect both Asian and African elephants, but lethal hemorrhagic disease is largely confined to Asian elephant calves and is predominantly associated with EEHV1. Milder disease caused by EEHV5 or EEHV4 is being increasingly recognized as well, but little is known about the latter, which is estimated to have diverged at least 35 million years ago from the others within a distinctive GC-rich branch of the Proboscivirus genus. Here, we have determined the complete genomic DNA sequence of a strain of EEHV4 obtained from a trunk wash sample collected from a surviving Asian elephant calf undergoing asymptomatic shedding during convalescence after an acute hemorrhagic disease episode. This represents the first example from among the three known GC-rich branch Proboscivirus species to be assembled and fully annotated. Several distinctive features of EEHV4 compared to AT-rich branch genomes are described.


Fecal diacylglycerol concentrations and calcium supplementation.
Growth factors are known to stimulate colonic proliferation via activation of protein kinase C by production of diacylglycerol (DAG) from membrane phosphatidyl inositol. Previous studies from our laboratories have shown that fecal bacteria can produce and metabolize DAG and that DAG can be absorbed by colonocytes, and thus might contribute to neoplasia. Calcium is a putative chemopreventive agent, and we have shown that calcium administration reduces fecal DAG concentrations, as well as rectal proliferation in patients after jejuno-ileal bypass surgery. The present study in normal volunteers eating self-selected diets demonstrates that fecal DAG concentrations are very constant with a coefficient of variation from 6.7 to 10.2%. Calcium administration showed a trend to reduce fecal DAG by 11% (P < 0.08). We conclude that fecal DAG levels can be determined in individuals on a self-selected diet from a single stool determination. If the trend to reduce fecal DAG by calcium is verified in more extensive studies, then the effects of calcium used in chemopreventive might, in part, reflect changes in the luminal lipid content.


The Relationship Between Urinary Total Polyphenols and the Frailty Phenotype in a Community-Dwelling Older Population: The InCHIANTI Study.
Frailty, an age-related state of increased vulnerability, is associated with a higher risk of multiple adverse events. Studies have suggested that the quality of dietary intake may affect the development of frailty. We hypothesized that frailty in older subjects would be associated with dietary total polyphenols (DTP) intake and its biomarker, urinary total polyphenols (UTP). The Invecchiare in Chianti (InCHIANTI) Study is a prospective cohort study set in the Chianti area (Italy). We used data at baseline from 811 participants aged 65 years and older. UTP was determined using the Folin-Ciocalteu assay after solid-phase extraction. DTP was estimated using a validated Food Frequency Questionnaire and our own polyphenol database. The frailty, prefrailty, and nonfrailty states were defined according to the Fried and colleagues' criteria. Multinomial logistic regressions adjusted for potential confounders were used to assess the relationship between polyphenols and frailty. Both DTP and UTP concentrations progressively decrease from nonfrail to frail participants. Participants in the highest UTP tertile compared to those in the lowest tertile were significantly less likely to be both frail (odds ratio [OR] = 0.36 [0.14-0.88], p = .025) and prefrail (OR = 0.64 [0.42-0.98], p = .038). Exhaustion and slowness were the only individual frailty criteria significantly associated with UTP tertiles. No significant association was observed between frailty and DTP, after adjustment for covariates. High concentrations of UTP were associated with lower prevalence of frailty and prefrailty in an older community-dwelling population. A polyphenol-rich diet may protect against frailty in older persons. Our findings should be confirmed in longitudinal studies.


Abnormal N-Glycosylation of a Novel Missense Creatine Transporter Mutant, G561R, Associated with Cerebral Creatine Deficiency Syndromes Alters Transporter Activity and Localization.
Cerebral creatine deficiency syndromes (CCDSs) are caused by loss-of-function mutations in creatine transporter (CRT, SLC6A8), which transports creatine at the blood-brain barrier and into neurons of the central nervous system (CNS). This results in low cerebral creatine levels, and patients exhibit mental retardation, poor language skills and epilepsy. We identified a novel human CRT gene missense mutation (c.1681 G>C, G561R) in Japanese CCDSs patients. The purpose of the present study was to evaluate the reduction of creatine transport in G561R-mutant CRT-expressing 293 cells, and to clarify the mechanism of its functional attenuation. G561R-mutant CRT exhibited greatly reduced creatine transport activity compared to wild-type CRT (WT-CRT) when expressed in 293 cells. Also, the mutant protein is localized mainly in intracellular membrane fraction, while WT-CRT is localized in plasma membrane. Western blot analysis revealed a 68 kDa band of WT-CRT protein in plasma membrane fraction, while G561R-mutant CRT protein predominantly showed bands at 55, 110 and 165 kDa in crude membrane fraction. The bands of both WT-CRT and G561R-mutant CRT were shifted to 50 kDa by N-glycosidase treatment. Our results suggest that the functional impairment of G561R-mutant CRT was probably caused by incomplete N-linked glycosylation due to misfolding during protein maturation, leading to oligomer formation and changes of cellular localization.


Small-conductance Ca(2+)-dependent K+ channels activated by ATP in murine colonic smooth muscle.
The patch-clamp technique was used to determine the ionic conductances activated by ATP in murine colonic smooth muscle cells. Extracellular ATP, UTP, and 2-methylthioadenosine 5'-triphosphate (2-MeS-ATP) increased outward currents in cells with amphotericin B-perforated patches. ATP (0.5-1 mM) did not affect whole cell currents of cells dialyzed with solutions containing ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. Apamin (3 x 10(-7) M) reduced the outward current activated by ATP by 32 +/- 5%. Single channel recordings from cell-attached patches showed that ATP, UTP, and 2-MeS-ATP increased the open probability of small-conductance, Ca(2+)-dependent K+ channels with a slope conductance of 5.3 +/- 0.02 pS. Caffeine (500 microM) enhanced the open probability of the small-conductance K+ channels, and ATP had no effect after caffeine. Pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid tetrasodium (PPADS, 10(-4) M), a nonselective P2 receptor antagonist, prevented the increase in open probability caused by ATP and 2-MeS-ATP. PPADS had no effect on the response to caffeine. ATP-induced hyperpolarization in the murine colon may be mediated by P2y-induced release of Ca2+ from intracellular stores and activation of the 5.3-pS Ca(2+)-activated K+ channels.


Audiovisual links in exogenous covert spatial orienting.
Subjects judged the elevation (up vs. down, regardless of laterality) of peripheral auditory or visual targets, following uninformative cues on either side with an intermediate elevation. Judgements were better for targets on either modality when preceded by an uninformative auditory cue on the side of the target. Experiment 2 ruled out nonattentional accounts for these spatial cuing effects. Experiment 3 found that visual cues affected elevation judgments for visual but not auditory targets. Experiment 4 confirmed that the effect on visual targets was attentional. In Experiment 5, visual cues produced spatial cuing when targets were always auditory, but saccades toward the cue may have been responsible. No such visual-to-auditory cuing effects were found in Experiment 6 when saccades were prevented, though they were present when eye movements were not monitored. These results suggest a one-way cross-modal dependence in exogenous covert orienting whereby audition influences vision, but not vice versa. Possible reasons for this asymmetry are discussed in terms of the representation of space within the brain.


Development and performance evaluation of a multi-PID muscle loading driven in vitro active-motion shoulder simulator and application to assessing reverse total shoulder arthroplasty.
In vitro active shoulder motion simulation can provide improved understanding of shoulder biomechanics; however, accurate simulators using advanced control theory have not been developed. Therefore, our objective was to develop and evaluate a simulator which uses real-time kinematic feedback and closed loop proportional integral differential (PID) control to produce motion. The simulator's ability to investigate a clinically relevant variable-namely muscle loading changes resulting from reverse total shoulder arthroplasty (RTSA)-was evaluated and compared to previous findings to further demonstrate its efficacy. Motion control of cadaveric shoulders was achieved by applying continuously variable forces to seven muscle groups. Muscle forces controlling each of the three glenohumeral rotational degrees of freedom (DOF) were modulated using three independent PID controllers running in parallel, each using measured Euler angles as their process variable. Each PID controller was configured and tuned to control the loading of a set of muscles which, from previous in vivo investigations, were found to be primarily responsible for movement in the PID's DOF. The simulator's ability to follow setpoint profiles for abduction, axial rotation, and horizontal extension was assessed using root mean squared error (RMSE) and average standard deviation (ASD) for multiple levels of arm mass replacement. A specimen was then implanted with an RTSA, and the effect of joint lateralization (0, 5, 10 mm) on the total deltoid force required to produce motion was assessed. Maximum profiling error was <2.1 deg for abduction and 2.2 deg for horizontal extension with RMSE of <1 deg. The nonprofiled DOF were maintained to within 5.0 deg with RMSE <1.0 deg. Repeatability was high, with ASDs of <0.31 deg. RMSE and ASD were similar for all levels of arm mass replacement (0.73-1.04 and 0.14-0.22 deg). Lateralizing the joint's center of rotation (CoR) increased total deltoid force by up to 8.5% body weight with the maximum early in abduction. This simulator, which is the first to use closed loop control, accurately controls the shoulder's three rotational DOF with high repeatability, and produces results that are in agreement with previous investigations. This simulator's improved performance, in comparison to others, increases the statistical power of its findings and thus its ability to provide new biomechanical insights.


Examining Human-Smartphone Interaction as a Proxy for Circadian Rhythm in Patients With Insomnia: Cross-Sectional Study.
The sleep and circadian rhythm patterns associated with smartphone use, which are influenced by mental activities, might be closely linked to sleep quality and depressive symptoms, similar to the conventional actigraphy-based assessments of physical activity. The primary objective of this study was to develop app-defined circadian rhythm and sleep indicators and compare them with actigraphy-derived measures. Additionally, we aimed to explore the clinical correlations of these indicators in individuals with insomnia and healthy controls. The mobile app "Rhythm" was developed to record smartphone use time stamps and calculate circadian rhythms in 33 patients with insomnia and 33 age- and gender-matched healthy controls, totaling 2097 person-days. Simultaneously, we used standard actigraphy to quantify participants' sleep-wake cycles. Sleep indicators included sleep onset, wake time (WT), wake after sleep onset (WASO), and the number of awakenings (NAWK). Circadian rhythm metrics quantified the relative amplitude, interdaily stability, and intradaily variability based on either smartphone use or physical activity data. Comparisons between app-defined and actigraphy-defined sleep onsets, WTs, total sleep times, and NAWK did not reveal any significant differences (all P>.05). Both app-defined and actigraphy-defined sleep indicators successfully captured clinical features of insomnia, indicating prolonged WASO, increased NAWK, and delayed sleep onset and WT in patients with insomnia compared with healthy controls. The Pittsburgh Sleep Quality Index scores were positively correlated with WASO and NAWK, regardless of whether they were measured by the app or actigraphy. Depressive symptom scores were positively correlated with app-defined intradaily variability (β=9.786, SD 3.756; P=.01) and negatively correlated with actigraphy-based relative amplitude (β=-21.693, SD 8.214; P=.01), indicating disrupted circadian rhythmicity in individuals with depression. However, depressive symptom scores were negatively correlated with actigraphy-based intradaily variability (β=-7.877, SD 3.110; P=.01) and not significantly correlated with app-defined relative amplitude (β=-3.859, SD 12.352; P=.76). This study highlights the potential of smartphone-derived sleep and circadian rhythms as digital biomarkers, complementing standard actigraphy indicators. Although significant correlations with clinical manifestations of insomnia were observed, limitations in the evidence and the need for further research on predictive utility should be considered. Nonetheless, smartphone data hold promise for enhancing sleep monitoring and mental health assessments in digital health research.


An investigation into symptoms, diagnosis, treatment, and treatment complications in patients with retrosternal goiter.
Retrosternal goiter refers to any thyroid enlargement in which over 50% of the thyroid permanently located under the thoracic inlet or the lower pole of thyroid is not palpable with the neck in hyperextended position. Due to the increasing number of surgical procedures of retrosternal goiter, the present study was carried out to examine the symptoms, diagnosis, treatment, and treatment complications in retrosternal goiter patients. Data related to demographic data (age and gender), clinical symptoms (dyspnea, dysphagia, dysphonia, lumps in neck, and hoarseness), methods of diagnosis (computed tomography [CT], chest X-ray [CXR], ultrasonography, and magnetic resonance imaging), and postoperative complications (bleeding, early and late dysphonia, early and late dyspnea, transient and permanent hypocalcemia, transient, and permanent recurrent laryngeal nerve paralysis) were collected. According to the results 71.4% of patients were women and most of the participants (67.1%) aged 45-60 years. Mass in the neck was the most frequent symptoms before surgery (88.6%). The most common incision for thyroidectomy (95/7%) was neck Collar incision. Diagnosis method in 82.9% and 17.1% of cases was, respectively, based on CT scans with CXR and CT scans with CXR and ultrasound. According to the postoperative pathologic findings, 58.5% of the cases were multinodular goiter, 22.9% were papillary cell carcinoma, 7.1% were medullary carcinoma, 5.7% were anaplastic carcinoma, 5.7% were thyroid lymphoma, and only 1.4% were thyroid adenoma. Postoperative complications occurred in 47.14% of patients. Most common complication was early transient dysphonia. This study recommends that retrosternal goiter should be operated early under suitable conditions, and the best diagnosis tool and best surgery methods are CT scan and surgery with collar incision, respectively.


[Educational level of mothers and their knowledge, attitude and practices concerning respiratory infections of their children].
An evaluation was performed on the influence of different variables on the knowledge, attitudes, and practices that mothers of children under the age of 5 have concerning acute respiratory infections (ARIs). Two groups of mothers were interviewed: 1) mothers of children seen in a polyclinic in the province of Havana (n = 221) and 2) mothers whose children were hospitalized in the respiratory unit of a pediatric hospital in the city of Havana (n = 200). The five variables analyzed were the educational level of the mother, her age, place of residence, number of children, and whether or not she was employed in the health service sector, as well as the interactions between the first variable and the other four variables. A questionnaire was used to measure the ARI knowledge, attitudes, and practices (KAP) of the mothers. Two levels of KAP sufficiency were taken into account, "Adequate I" for obtaining 70% or more of the maximum possible score and "Adequate II" for achieving at least a 50% score. For both levels of adequacy, the KAP of the mothers working in the health sector was significantly higher than that of the mothers interviewed in the secondary health care center. This suggests that the poorer knowledge, attitudes, and practices of mothers with hospitalized children may have contributed to the children's hospitalization. The most important variable associated with sufficient knowledge was educational level, which highlights the fundamental and positive influence education exerts on the preventive and curative care that mothers provide their children with ARI.


The role of laparoscopic approach for anastomotic leakage after minimally invasive surgery for colorectal cancer.
The objectives of this study were to evaluate the feasibility and safety of a re-laparoscopic approach to manage anastomotic leakage after minimally invasive colorectal resection and to compare its clinical outcomes with those obtained using an open approach. We retrospectively reviewed clinical data from 1714 patients who underwent colorectal cancer resection from September 2006 to August 2009 at the Korea University Medical Center. Clinical data from a total of 57 surgery patients who developed anastomotic leakage were analyzed. Twenty-six leakage cases were managed laparoscopically, whereas the remaining 31 leakage cases were managed using an open approach. There were no significant differences in age, sex, or other clinical features between patients in the two groups. The total operation time was shorter in the laparoscopic group (107.3 ± 68.1 minutes) than in the open group (126.5 ± 50.1 minutes), but this difference was not statistically significant (P = .230). Six cases in each group required additional procedures such as reoperation or percutaneous intervention (P = .126). There was one case of postoperative mortality in the open group. Median (quartiles 25%-75%) number of days required to resume a soft diet tended to be shorter in the laparoscopic group than the open group (5 [3-7] versus 6 [5-10] days; P = .057). Patients in both groups showed similar postoperative complications including intraabdominal abscess; however, the incidence of wound infection was significantly lower in the laparoscopic group than the open group (3.8% versus 25.8%; P = .031). Compared with conventional open treatment of anastomotic leakage, the laparoscopic approach resulted in fewer wound complications and tendency of early recovery of bowel movement without an increase in adverse outcomes. Using a laparoscopic approach, all the advantages of minimally invasive surgery can be realized in patients who develop anastomotic leakage after minimally invasive surgery.


Changes in pain intensity and health related quality of life with Iyengar yoga in nonspecific chronic low back pain: A randomized controlled study.
Nonspecific chronic low back (nCLBP) pain is prevalent among adult population and often leads to functional limitations, psychological symptoms, lower quality of life (QOL), and higher healthcare costs. The purpose of this study was to determine the efficacy of Iyengar yoga therapy on pain intensity and health related quality of life (HRQOL) with nCLBP. To compare the effect of Iyengar yoga therapy and conventional exercise therapy on pain intensity and HRQOL in nonspecific chronic low back pain. Experimental study with random sampling technique. Sixty subjects who fulfilled the selection criteria were randomly assigned to Iyengar yoga (yoga group, n = 30) and control group (exercise group, n = 30). Participants completed low back pain evaluation form and HRQOL-4 questionnaire before their intervention and again 4 weeks and 6 month later. Yoga group underwent 29 yogic postures training and exercise group had undergone general exercise program for 4 weeks. Repeated measures analysis of variance (ANOVA) was used to analyze group differences over time, while controlling for baseline differences. Patients in both groups experienced significant reduction in pain and improvement in HRQOL. In visual analogue scale (VAS) yoga group showed reduction of 72.81% (P = 0.001) as compared to exercise group 42.50% (P = 0.001). In HRQOL, yoga group showed reduction of 86.99% (P = 0.001) as compared to exercise group 67.66% (P = 0.001). These results suggest that Iyengar yoga provides better improvement in pain reduction and improvement in HRQOL in nonspecific chronic back pain than general exercise.


A family of neofunctionalized Ty3/gypsy retrotransposon genes in mammalian genomes.
A family of at least eleven genes called Mar related to long terminal repeat retrotransposons from the Ty3/gypsy group, including two genes previously identified as such, is present in human and mouse genomes. Single orthologous copies were identified for most Mar genes in different mammals. All of them have lost essential structural features necessary for autonomous retrotransposition before divergence between mouse and human. Three Mar genes also have introns at identical positions in human and mouse. Hence, Mar genes do not correspond to functional retrotransposons. Mar genes evolved under purifying selection, strongly suggesting that they are not pseudogenic relics but rather neofunctionalized retrotransposon genes. All putative Mar proteins display sequence similarity to the capsid-like domain of the Gag protein of Tf1/Sushi retrotransposons. In addition, three Mar proteins have conserved the Gag CCHC zinc finger motif, suggesting a role in nucleic acid binding. Some Mar genes have also retained from their retrotransposon origin a -1 ribosomal frameshifting between the gag-related open reading frame and a region encoding a putative aspartyl protease domain. EST analysis revealed that the majority of Mar genes are expressed in brain as well as in other tissues and organs. Some Mar proteins might function as transcription factors or be involved in the control of cell proliferation and apoptosis. Strikingly, as many as eight Mar genes are located on the X chromosome in human, mouse and other mammals, and at least two of the autosomal genes are subject to imprinting. We suggest that retrotransposons might be a source for epigenetically regulated genes. Epigenetic regulation of these neogenes might be derived from the cellular defense mechanisms having controlled their retrotransposon ancestor.


Entrapment of Src protein tyrosine kinase in sugar-modified silica.
A novel sugar-modified silica has been used to entrap for the first time a protein tyrosine kinase (PTK). Silane precursors bearing covalently attached gluconamide moieties were used in combination with the biocompatible precursor diglycerylsilane (DGS) to generate sol-gel derived silica that was able to encapsulate highly active Src PTK and preserve the activity of the enzyme over multiple uses. The relative activity of the enzyme was assayed using a LANCE based fluorescence resonance energy transfer method involving time-gated detection of fluorescence from a europium labeled antiphosphotyrosine antibody and Cy5 labeled streptavidin upon mutual binding to biotinylated phosphopeptides. Using this detection method, with the antibody and streptavidin external to the sol-gel matrix, it was possible to detect the phosphorylation of peptides with molecular weights of up to 2300 Da using the entrapped enzyme in N-(3-triethoxysilylpropyl)gluconamide (GLTES) doped glasses. Src kinase-doped glasses, derived from precursors such as tetramethyl orthosilicate, tetraethyl orthosilicate, or DGS that did not contain GLTES, provided no detectable enzyme activity. The addition of 1 mM ATP to the GLTES/DGS sol before the encapsulation of the protein increased the activity of the enzyme in the resulting gel, likely through a ligand-based stabilization mechanism. The use of such a system for determination of PTK activity and inhibition is demonstrated, setting the stage for the development of chromatographic and microarray based methods for the screening of kinase inhibitors.


O acetylation of the enterobacterial common antigen polysaccharide is catalyzed by the product of the yiaH gene of Escherichia coli K-12.
The carbohydrate component of the enterobacterial common antigen (ECA) of Escherichia coli K-12 occurs primarily as a water-soluble cyclic polysaccharide located in the periplasm (ECA(CYC)) and as a phosphoglyceride-linked linear polysaccharide located on the cell surface (ECA(PG)). The polysaccharides of both forms are comprised of the amino sugars N-acetyl-D-glucosamine (GlcNAc), N-acetyl-D-mannosaminuronic acid (ManNAcA), and 4-acetamido-4,6-dideoxy-D-galactose (Fuc4NAc). These amino sugars are linked to one another to form trisaccharide repeat units with the structure -->3-alpha-D-Fuc4NAc-(1-->4)-beta-D-ManNAcA-(1-->4)-alpha-D-GlcNAc-(1-->. The hydroxyl group in the 6 position of the GlcNAc residues of both ECA(CYC) and ECA(PG) are nonstoichiometrically esterified with acetyl groups. Random transposon insertion mutagenesis of E. coli K-12 resulted in the generation of a mutant defective in the incorporation of O-acetyl groups into both ECA(CYC) and ECA(PG). This defect was found to be due to an insertion of the transposon into the yiaH locus, a putative gene of unknown function located at 80.26 min on the E. coli chromosomal map. Bioinformatic analyses of the predicted yiaH gene product indicate that it is an integral inner membrane protein that is a member of an acyltransferase family of enzymes found in a wide variety of organisms. The results of biochemical and genetic experiments presented here strongly support the conclusion that yiaH encodes the O-acetyltransferase responsible for the incorporation of O-acetyl groups into both ECA(CYC) and ECA(PG). Accordingly, we propose that this gene be designated wecH.


Methods to improve patient recruitment and retention in stroke trials.
The success of randomized-controlled stroke trials is dependent on the recruitment and retention of a sufficient number of patients, but fewer than half of all trials meet their target number of patients. We performed a search and review of the literature, and conducted a survey and workshop among 56 European stroke trialists, to identify barriers, suggest methods to improve recruitment and retention, and make a priority list of interventions that merit further evaluation. The survey and workshop identified a number of barriers to patient recruitment and retention, from patients' incapacity to consent, to handicaps that prevent patients from participation in trial-specific follow-up. Methods to improve recruitment and retention may include simple interventions with individual participants, funding of research networks, and reimbursement of new treatments by health services only when delivered within clinical trials. The literature review revealed that few methods have been formally evaluated. The top five priorities for evaluation identified in the workshop were as follows: short and illustrated patient information leaflets, nonwritten consent, reimbursement for new interventions only within a study, and monetary incentives to institutions taking part in research (for recruitment); and involvement of patient groups, remote and central follow-up, use of mobile devices, and reminders to patients about their consent to participate (for retention). Many interventions have been used with the aim of improving recruitment and retention of patients in stroke studies, but only a minority has been evaluated. We have identified methods that could be tested, and propose that such evaluations may be nested within on-going clinical trials.


Selective Delivery of PEGylated Compounds to Tumor Cells by Anti-PEG Hybrid Antibodies.
Polyethylene glycol (PEG) is attached to many peptides, proteins, liposomes, and nanoparticles to reduce their immunogenicity and improve their pharmacokinetic and therapeutic properties. Here, we describe hybrid antibodies that can selectively deliver PEGylated medicines, imaging agents, or nanomedicines to target cells. Human IgG1 hybrid antibodies αPEG:αHER2 and αPEG:αCD19 were shown by ELISA, FACS, and plasmon resonance to bind to both PEG and HER2 receptors on SK-BR-3 breast adenocarcinoma and BT-474 breast ductal carcinoma cells or CD19 receptors on Ramos and Raji Burkitt's lymphoma cells. In addition, αPEG:αHER2 specifically targeted PEGylated proteins, liposomes, and nanoparticles to SK-BR-3 cells that overexpressed HER2, but not to HER2-negative MCF-7 breast adenocarcinoma cells. Endocytosis of PEGylated nanoparticles into SK-BR-3 cells was induced specifically by the αPEG:αHER2 hybrid antibody, as observed by confocal imaging of the accumulation of Qdots inside SK-BR-3 cells. Treatment of HER2(+) SK-BR-3 and BT-474 cancer cells with αPEG:αHER2 and the clinically used chemotherapeutic agent PEGylated liposomal doxorubicin for 3 hours enhanced the in vitro effectiveness of PEGylated liposomal doxorubicin by over two orders of magnitude. Hybrid anti-PEG antibodies offer a versatile and simple method to deliver PEGylated compounds to cellular locations and can potentially enhance the therapeutic efficacy of PEGylated medicines.


An E-Platform for Rehabilitation of Persons With Hearing Problems.
The aim of this research forum article was to describe a feasible web-based solution for improving the quality of life of persons with hearing problems, such as hearing loss or tinnitus. The online platform was developed at the Department of Behavioural Sciences and Learning at Linköping University, Sweden, and has been running for a number of years and used in numerous studies and treatments. The security aspects of the platform as well as the process flow for running a study or treatment are described, focusing primarily on the technical and practical considerations. Also presented are the design characteristics and the main features and functions available in the platform. We point out the many advantages of running Internet-assisted intervention treatments, the challenges that we have faced, and some intended developments. Many of our research colleagues, both from Sweden and other countries, have already implemented or intend to implement their own studies on this platform. Audiological rehabilitation can be delivered via the Internet using a stable online platform. Security and usability are important factors to have in mind for the design as well as adaptability to the patients. A next development step is to implement and test blended treatments using video conferencing inside the platform.


Physical activity and risks of proximal and distal colon cancers: a systematic review and meta-analysis.
Although there is convincing epidemiological evidence that physical activity is associated with a reduced risk of colon cancer, it is unclear whether physical activity is differentially associated with the risks of proximal colon and distal colon cancers. We conducted a systematic review and meta-analysis to investigate this issue. MEDLINE and EMBASE were searched for English-language cohort and case-control studies that examined associations between physical activity and the risks of proximal colon and distal colon cancers. A random-effects meta-analysis was conducted to estimate the summary relative risks (RRs) for the associations between physical activity and the risks of the two cancers. All statistical tests were two-sided. A total of 21 studies met the inclusion criteria. The summary relative risk of the main results from these studies indicated that the risk of proximal colon cancer was 27% lower among the most physically active people compared with the least active people (RR = 0.73, 95% confidence interval [CI] = 0.66 to 0.81). An almost identical result was found for distal colon cancer (RR = 0.74, 95% CI = 0.68 to 0.80). The results of this systematic review and meta-analysis suggest that physical activity is associated with a reduced risk of both proximal colon and distal colon cancers, and that the magnitude of the association does not differ by subsite. Given this finding, future research on physical activity and colon cancer should focus on other aspects of the association that remain unclear, such as whether sedentary behavior and nonaerobic physical activity are associated with the risk of colon cancer.


Enteric neural crest cells regulate vertebrate stomach patterning and differentiation.
In vertebrates, the digestive tract develops from a uniform structure where reciprocal epithelial-mesenchymal interactions pattern this complex organ into regions with specific morphologies and functions. Concomitant with these early patterning events, the primitive GI tract is colonized by the vagal enteric neural crest cells (vENCCs), a population of cells that will give rise to the enteric nervous system (ENS), the intrinsic innervation of the GI tract. The influence of vENCCs on early patterning and differentiation of the GI tract has never been evaluated. In this study, we report that a crucial number of vENCCs is required for proper chick stomach development, patterning and differentiation. We show that reducing the number of vENCCs by performing vENCC ablations induces sustained activation of the BMP and Notch pathways in the stomach mesenchyme and impairs smooth muscle development. A reduction in vENCCs also leads to the transdifferentiation of the stomach into a stomach-intestinal mixed phenotype. In addition, sustained Notch signaling activity in the stomach mesenchyme phenocopies the defects observed in vENCC-ablated stomachs, indicating that inhibition of the Notch signaling pathway is essential for stomach patterning and differentiation. Finally, we report that a crucial number of vENCCs is also required for maintenance of stomach identity and differentiation through inhibition of the Notch signaling pathway. Altogether, our data reveal that, through the regulation of mesenchyme identity, vENCCs act as a new mediator in the mesenchymal-epithelial interactions that control stomach development.


Cue-induced food seeking after punishment is associated with increased Fos expression in the lateral hypothalamus and basolateral and medial amygdala.
In humans, relapse to unhealthy eating habits following dieting is a significant impediment to obesity treatment. Food-associated cues are one of the main triggers of relapse to unhealthy eating during self-imposed abstinence. Here we report a behavioral method examining cue-induced relapse to food seeking following punishment-induced suppression of food taking. We trained male rats to lever press for food pellets that were delivered after a 10-s conditional stimulus (CS) (appetitive). Following training, 25% of reinforced lever presses resulted in the presentation of a compound stimulus consisting of a novel CS (aversive) and the appetitive CS followed by a pellet and footshock. After punishment-imposed abstinence, we tested the rats in an extinction test where lever pressing resulted in the presentation of either the appetitive or aversive CS. We then compared activity of lateral hypothalamus (LH) and associated extrahypothalamic regions following this test. We also assessed Fos expression in LH orexin and GABA neurons. We found that cue-induced relapse of food seeking on test was higher in rats tested with the appetitive CS compared to the aversive CS. Relapse induced by the appetitive CS was associated with increased Fos expression in LH, caudal basolateral amygdala (BLA), and medial amygdala (MeA). This relapse was also associated with increased Fos expression in LH orexin and VGAT-expressing neurons. These data show that relapse to food seeking can be induced by food-associated cues after punishment-imposed abstinence, and this relapse is associated with increased activity in LH, caudal BLA, and MeA. (PsycINFO Database Record


Histochemical and immunocytochemical compartments of the thalamic VPM nucleus in monkeys and their relationship to the representational map.
The ventral posteromedial nucleus (VPM) of the monkey thalamus was investigated with correlative anatomical and physiological techniques. On the basis of staining for cytochrome oxidase (CO), VPM is divided into a lightly stained, background matrix domain and an intensely stained rod domain. The latter consists of elongated rods of large, medium, and small cells, 500 microns wide on average and extending anteroposteriorly, many of them through the full extent of the nucleus. The matrix, consisting of small cells, penetrates between the rods and expands at the dorsomedial, ventrolateral, and posterior aspects of VPM. Multiunit mapping reveals that VPM contains a dorsally situated representation of the contralateral side of the head, face, eye, and interior of the mouth and a medially situated representation of the ipsilateral side of the lips and interior of the mouth, and that the same small region is represented in the same relative position through the full anteroposterior extent of the nucleus. Earlier work had shown that single CO rods contain the representation of the same portion of the periphery throughout their length. The present study suggests that rods in equivalent positions may represent the same portion of the periphery from animal to animal. The cells of the rod and matrix domains show different patterns of immunoreactivity. Virtually all of the large- and medium-sized rod cells are immunoreactive for the calcium-binding protein parvalbumin, and many are stained by the monoclonal antibody CAT 301. Small GABA-immunoreactive cells and terminal-like puncta are highly concentrated in the rods but are dispersed in the matrix. In the matrix, all non-GABA cells are small, immunoreactive for 28-kDa calbindin, and not stained by CAT 301. They appear to form part of a wider system of calbindin-positive cells that extends into adjacent nuclei. The CO rods are indicative of the modularity of the lemniscal component of the trigeminal part of the somatic sensory system at thalamic levels. Thalamocortical relay neurons in this compartment of VPM express a calcium-binding protein and a surface proteoglycan that distinguishes them from relay neurons in the matrix compartment of the nucleus. In the following paper (Rausell and Jones, 1991), the rod and matrix compartments are shown also to have different patterns of input and output connections.


KIT pathway alterations in mucosal melanomas of the vulva and other sites.
A significant proportion of mucosal melanomas contain alterations in KIT. The aim of this study was to characterize the pattern of KIT, NRAS, and BRAF mutations in mucosal melanomas at specific sites and to assess activation of the KIT downstream RAF/MEK/extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K)/AKT pathways in mucosal melanoma specimens. Seventy-one primary mucosal melanomas from various sites were studied. Mutation analysis was done by DNA sequencing. Expression of KIT, phosphorylated (p)-ERK, and p-AKT was evaluated by immunohistochemistry. KIT mutations were detected in 35% (8 of 23) of vulvar, 9% (2 of 22) of anorectal, 7% (1 of 14) of nasal cavity, and 20% (1 of 5) of penile melanomas. No KIT mutations were found in 7 vaginal melanomas. The difference in KIT mutation frequency between vulvar and nonvulvar cases was statistically significant (P = 0.014). The overall frequencies of NRAS and BRAF mutations were 10% and 6%, respectively. Notably, vaginal melanomas showed a NRAS mutation rate of 43%. KIT gene amplification (≥4 copies), as assessed by quantitative real-time PCR, was observed in 19% of cases. KIT expression was associated with KIT mutation status (P < 0.001) and was more common in vulvar than nonvulvar tumors (P = 0.016). Expression of p-ERK and p-AKT was observed in 42% and 59% of tumors, respectively, and occurred irrespective of KIT/NRAS/BRAF mutation status. NRAS mutation was associated with worse overall survival in univariate analysis. Results show that KIT mutations are more common in vulvar melanomas than other types of mucosal melanomas and that both the RAF/MEK/ERK and PI3K/AKT pathways are activated in mucosal melanoma specimens.


Emotion regulation and relations to well-being across the lifespan.
The specific strategies that individuals use to regulate their emotions have shown strong associations with various indices of well-being. However, theoretical accounts suggest that strategy use, and the associations between strategy use and well-being, may change across the life span. Attempts have been made to assess whether levels of strategy use, and the association between strategy use and well-being, change across development; however, studies typically do not take into account potential differences in base rates of item endorsement across the life span. Therefore, the current study had two objectives. First, we sought to examine whether relative ER strategy reliance, or the proportional degree to which an individual relies on various ER strategies, varied across three developmental periods: early adolescence, young adulthood, and adulthood. Second, we sought to identify whether the associations between relative strategy use and well-being differed significantly across these developmental periods. Results showed that relative reliance on distraction, rumination, relaxation, and suppression differed significantly across age groups. Moreover, results showed that the association between relative use of reappraisal and suppression interacted with age group to predict relationship quality. Relative strategy use and age did not interact significantly to predict anxiety and depressive symptoms. Theoretical and practical implications are discussed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Modeling drug-resistant tuberculosis amplification rates and intervention strategies in Bangladesh.
Tuberculosis (TB) is the seventh leading cause of morbidity and mortality in Bangladesh. Although the National TB control program (NTP) of Bangladesh is implementing its nationwide TB control strategies, more specific and effective single or combination interventions are needed to control drug-susceptible (DS) and multi-drug resistant (MDR) TB. In this study, we developed a two strain TB mathematical model with amplification and fit it to the Bangladesh TB data to understand the transmission dynamics of DS and MDR TB. Sensitivity analysis was used to identify important parameters. We evaluated the cost-effectiveness of varying combinations of four basic control strategies including distancing, latent case finding, case holding and active case finding, all within the optimal control framework. From our fitting, the model with different transmission rates between DS and MDR TB best captured the Bangladesh TB reported case counts. The estimated basic reproduction number for DS TB was 1.14 and for MDR TB was 0.54, with an amplification rate of 0.011 per year. The sensitivity analysis also indicated that the transmission rates for both DS and MDR TB had the largest influence on prevalence. To reduce the burden of TB (both DS and MDR), our finding suggested that a quadruple control strategy that combines distancing control, latent case finding, case holding and active case finding is the most cost-effective. Alternative strategies can be adopted to curb TB depending on availability of resources and policy makers' decisions.


Nutritional Wheat Amylase-Trypsin Inhibitors Promote Intestinal Inflammation via Activation of Myeloid Cells.
Wheat amylase-trypsin inhibitors (ATIs) are nutritional activators of innate immunity, via activation of the toll-like receptor 4 (TLR4) on myeloid cells. We aimed to characterize the biologic activity of ATIs in various foods and their effect on intestinal inflammation. We selected 38 different gluten-containing and gluten-free products, either unprocessed (such as wheat, rye, barley, quinoa, amaranth, soya, lentils, and rice) or processed (such as pizza, pasta, bread, and biscuits). ATIs were extracted and their biological activities determined in TLR4-responsive mouse and human cell lines. Effects of oral ATIs on intestinal inflammation were determined in healthy C57BL/6 mice on a gluten-free or ATI-free diet and in mice given low-level polyinosinic:polycytidylic acid or dextran sodium sulfate to induce colitis. Parameters of innate and adaptive immune activation were determined in duodenum, ileum, colon, and mesenteric lymph nodes. Modern gluten-containing staples had levels of TLR4-activating ATIs that were as much as 100-fold higher than in most gluten-free foods. Processed or baked foods retained ATI bioactivity. Most older wheat variants (such as Emmer or Einkorn) had lower bioactivity than modern (hexaploid) wheat. ATI species CM3 and 0.19 were the most prevalent activators of TLR4 in modern wheat and were highly resistant to intestinal proteolysis. Their ingestion induced modest intestinal myeloid cell infiltration and activation, and release of inflammatory mediators-mostly in the colon, then in the ileum, and then in the duodenum. Dendritic cells became prominently activated in mesenteric lymph nodes. Concentrations of ATIs found in a normal daily gluten-containing diet increased low-level intestinal inflammation. Gluten-containing cereals have by far the highest concentrations of ATIs that activate TLR4. Orally ingested ATIs are largely resistant to proteases and heat, and increase intestinal inflammation by activating gut and mesenteric lymph node myeloid cells.


A subpopulation of tegument protein vhs localizes to detergent-insoluble lipid rafts in herpes simplex virus-infected cells.
Virion host shutoff (vhs) is a 58-kDa protein encoded by the UL41 gene of herpes simplex virus (HSV). vhs resides within the tegument of HSV, enters the cell cytoplasm at infection, and destabilizes host cell and viral mRNA. Late in infection, vhs must be assembled into the tegument of progeny virions, a poorly understood process. Using an anti-vhs antiserum and Western blotting of total cell or cytoplasmic extracts, we found that vhs is largely insoluble in HSV-infected cells, even in the presence of high levels of salt and the detergent Triton X-100. Furthermore, a subpopulation of vhs appears to be associated with detergent-insoluble lipid rafts and this raft population is enriched in a cytoplasmic fraction which contains assembling and mature HSV particles. Our data raise the possibility that HSV tegument polypeptides associate with membrane rafts, in common with the matrix proteins of a number of other viruses.


Mystery shopping and alcohol sales: do supermarkets and liquor stores sell alcohol to underage customers?
The Dutch national policy regarding alcohol and youth relies on retailers' willingness to refuse to sell alcohol to underage customers. This study examined unobtrusively whether supermarkets and liquor stores do indeed comply with the legal age restrictions for alcohol sales. A research protocol was developed based on the methodology of mystery shopping. Using the protocol, 150 supermarkets and 75 liquor stores were visited by 15-year-old adolescents who tried to buy soft alcoholic beverages (legal age, 16 years), and 75 liquor stores were visited by 17-year-old adolescents who tried to buy strong alcoholic beverages (legal age, 18). Of all 300 buying attempts, 86% were successful. In supermarkets, 88% of all attempts succeeded. In liquor stores, a difference was found between the purchase of strong alcohol by 17-year-olds (89%) and the purchase of soft alcoholic beverages by 15-year-olds (77%). In only 71 of all visits, mystery shoppers were asked for an ID. In 39% of these cases, they were still able to buy alcohol. Female adolescents were more successful in buying alcohol than male adolescents. The results show that supermarkets and liquor stores generally fail to see the need for extra care when young customers try to buy alcohol. Legal age restrictions without enforcement and facilitation clearly do not suffice to protect adolescents from early exposure to alcohol.


ERK1/2 mediates insulin stimulation of Na(+),K(+)-ATPase by phosphorylation of the alpha-subunit in human skeletal muscle cells.
Insulin stimulates Na(+),K(+)-ATPase activity and induces translocation of Na(+),K(+)-ATPase molecules to the plasma membrane in skeletal muscle. We determined the molecular mechanism by which insulin regulates Na(+),K(+)-ATPase in differentiated primary human skeletal muscle cells (HSMCs). Insulin action on Na(+),K(+)-ATPase was dependent on ERK1/2 in HSMCs. Sequence analysis of Na(+),K(+)-ATPase alpha-subunits revealed several potential ERK phosphorylation sites. Insulin increased ouabain-sensitive (86)Rb(+) uptake and [(3)H]ouabain binding in intact cells. Insulin also increased phosphorylation and plasma membrane content of the Na(+),K(+)-ATPase alpha(1)- and alpha(2)-subunits. Insulin-stimulated Na(+),K(+)-ATPase activation, phosphorylation, and translocation of alpha-subunits to the plasma membrane were abolished by 20 microm PD98059, which is an inhibitor of MEK1/2, an upstream kinase of ERK1/2. Furthermore, inhibitors of phosphatidylinositol 3-kinase (100 nm wortmannin) and protein kinase C (10 microm GF109203X) had similar effects. Notably, insulin-stimulated ERK1/2 phosphorylation was abolished by wortmannin and GF109203X in HSMCs. Insulin also stimulated phosphorylation of alpha(1)- and alpha(2)-subunits on Thr-Pro amino acid motifs, which form specific ERK substrates. Furthermore, recombinant ERK1 and -2 kinases were able to phosphorylate alpha-subunit of purified human Na(+),K(+)-ATPase in vitro. In conclusion, insulin stimulates Na(+),K(+)-ATPase activity and translocation to plasma membrane in HSMCs via phosphorylation of the alpha-subunits by ERK1/2 mitogen-activated protein kinase.


Evaluating pre-pregnancy dietary diversity vs. dietary quality scores as predictors of gestational diabetes and hypertensive disorders of pregnancy.
Dietary diversity scores (DDS) are considered as metrics for monitoring the implementation of the UN's Sustainable Development Goals, but they need to be rigorously evaluated. To examine two DDS, the Food Groups Index (FGI), and the Minimum Dietary Diversity-Women (MDD-W), alongside two dietary quality scores, the Alternate Healthy Eating Index (AHEI-2010) and the Prime Diet Quality Score (PDQS), with risks of gestational diabetes mellitus (GDM) and hypertensive disorders of pregnancy (HDPs). The analysis included 21,312 (GDM) and 19,917 (HDPs) singleton births reported in the Nurses' Health Study II cohort (1991-2001), among women without major chronic disease or GDM/HDPs. Scores were derived using prepregnancy diet collected by a comprehensive food frequency questionnaire. Multivariable models were utilized to calculate relative risks (RR) and confidence intervals (95%CIs). Incident GDM (n = 916) and HDPs (n = 1,421) were reported. The MDD-W and FGI were not associated with risk of GDM or HDPs, but the AHEI-2010 and PDQS were associated with a lower risk of GDM and marginally lower risk of HDP. The RR's of GDM comparing the highest vs. lowest quintiles were 1.00 (95%CI: 0.79, 1.27; p-trend = 0.82) for MDD-W, 0.96 (95%CI: 0.76, 1.22; p-trend = 0.88) for FGI, 0.63 (95%CI: 0.50, 0.81; p-trend <0.0001) for the AHEI-2010 and 0.68 (95%CI: 0.54, 0.86; p-trend = 0.003) for the PDQS. Similarly, the RR's of HDPs were 0.92 (95%CI: 0.75, 1.12, p-trend = 0.94) for MDD-W, 0.97 (95%CI: 0.79, 1.17; p-trend = 0.83) for FGI, 0.84 (95%CI: 0.70, 1.02; p-trend = 0.07) for AHEI-2010 and 0.89 (95%CI: 0.74, 1.09; p-trend = 0.07) for PDQS. MDD-W and FGI did not predict the risk of GDM and HDPs. These DDS should not be widely used as metrics for achieving dietary goals in their present form. The Prime Diet Quality Score warrants further testing as a promising measure of a sustainable and healthy diet on a global scale.


Measuring plasma fibrinogen to predict stroke and myocardial infarction.
Epidemiological observations indicate that high plasma fibrinogen levels are strongly correlated with the frequency of two major thrombotic complications of atherosclerosis, stroke and myocardial infarction. Thrombosis is increasingly recognized as a central mechanism in stroke and myocardial infarction, and fibrinogen is involved in events thought to play a major role in thrombosis. Therefore, elucidation of the relationship between fibrinogen and thrombosis may strengthen the predictive value of this protein and suggest new treatment to prevent stroke and myocardial infarction. The current data relating fibrinogen to thrombosis are not easy to reconcile with the available epidemiological observations. In addition, advances in understanding the atherogenic potential of several risk factors for coronary heart disease have used information on the measurement of the risk factors in population-based studies. Thus, measuring plasma fibrinogen to predict stroke and myocardial infarction may be important in gaining insight into the thrombogenic potential of this protein and in inspiring new strategies against the thrombotic complications of atherosclerosis.


High resolution analysis of follicular lymphoma genomes reveals somatic recurrent sites of copy-neutral loss of heterozygosity and copy number alterations that target single genes.
A multiplatform approach, including conventional cytogenetic techniques, BAC array comparative genomic hybridization, and Affymetrix 500K SNP arrays, was applied to the study of the tumor genomes of 25 follicular lymphoma biopsy samples with paired normal DNA samples to characterize balanced translocations, copy number imbalances, and copy-neutral loss of heterozygosity (cnLOH). In addition to the t(14;18), eight unique balanced translocations were found. Commonly reported FL-associated copy number regions were revealed including losses of 1p32-36, 6q, and 10q, and gains of 1q, 6p, 7, 12, 18, and X. The most frequent regions affected by copy-neutral loss of heterozygosity were 1p36.33 (28%), 6p21.3 (20%), 12q21.2-q24.33 (16%), and 16p13.3 (24%). We also identified by SNP analysis, 45 aberrant regions that each affected one gene, including CDKN2A, CDKN2B, FHIT, KIT, PEX14, and PTPRD, which were associated with canonical pathways involved in tumor development. This study illustrates the power of using complementary high-resolution platforms on paired tumor/normal specimens and computational analysis to provide potential insights into the significance of single-gene somatic aberrations in FL tumorigenesis.