Pseudomonas aeruginosa-induced bleb-niche formation in epithelial cells is independent of actinomyosin contraction and enhanced by loss of cystic fibrosis transmembrane-conductance regulator osmoregulatory function.
The opportunistic pathogen Pseudomonas aeruginosa can infect almost any site in the body but most often targets epithelial cell-lined tissues such as the airways, skin, and the cornea of the eye. A common predisposing factor is cystic fibrosis (CF), caused by defects in the cystic fibrosis transmembrane-conductance regulator (CFTR). Previously, we showed that when P. aeruginosa enters epithelial cells it replicates intracellularly and occupies plasma membrane blebs. This phenotype is dependent on the type 3 secretion system (T3SS) effector ExoS, shown by others to induce host cell apoptosis. Here, we examined mechanisms for P. aeruginosa-induced bleb formation, focusing on its relationship to apoptosis and the CFTR. The data showed that P. aeruginosa-induced blebbing in epithelial cells is independent of actin contraction and is inhibited by hyperosmotic media (400 to 600 mOsM), distinguishing bacterially induced blebs from apoptotic blebs. Cells with defective CFTR displayed enhanced bleb formation upon infection, as demonstrated using bronchial epithelial cells from a patient with cystic fibrosis and a CFTR inhibitor, CFTR(Inh)-172. The defect was found to be correctable either by incubation in hyperosmotic media or by complementation with CFTR (pGFP-CFTR), suggesting that the osmoregulatory function of CFTR counters P. aeruginosa-induced bleb-niche formation. Accordingly, and despite their reduced capacity for bacterial internalization, CFTR-deficient cells showed greater bacterial occupation of blebs and enhanced intracellular replication. Together, these data suggest that P. aeruginosa bleb niches are distinct from apoptotic blebs, are driven by osmotic forces countered by CFTR, and could provide a novel mechanism for bacterial persistence in the host. Pseudomonas aeruginosa is an opportunistic pathogen problematic in hospitalized patients and those with cystic fibrosis (CF). Previously, we showed that P. aeruginosa can enter epithelial cells and replicate within them and traffics to the membrane blebs that it induces. This "bleb-niche" formation requires ExoS, previously shown to cause apoptosis. Here, we show that the driving force for bleb-niche formation is osmotic pressure, differentiating P. aeruginosa-induced blebs from apoptotic blebs. Either CFTR inhibition or CFTR mutation (as seen in people with CF) causes P. aeruginosa to make more bleb niches and provides an osmotic driving force for blebbing. CFTR inhibition also enhances bacterial occupation of blebs and intracellular replication. Since CFTR is targeted for removal from the plasma membrane when P. aeruginosa invades a healthy cell, these findings could relate to pathogenesis in both CF and healthy patient populations.


Pontellid copepods, Labidocera spp., affected by ocean acidification: A field study at natural CO2 seeps.
CO2 seeps in coral reefs were used as natural laboratories to study the impacts of ocean acidification on the pontellid copepod, Labidocera spp. Pontellid abundances were reduced by ∼70% under high-CO2 conditions. Biological parameters and substratum preferences of the copepods were explored to determine the underlying causes of such reduced abundances. Stage- and sex-specific copepod lengths, feeding ability, and egg development were unaffected by ocean acidification, thus changes in these physiological parameters were not the driving factor for reduced abundances under high-CO2 exposure. Labidocera spp. are demersal copepods, hence they live amongst reef substrata during the day and emerge into the water column at night. Deployments of emergence traps showed that their preferred reef substrata at control sites were coral rubble, macro algae, and turf algae. However, under high-CO2 conditions they no longer had an association with any specific substrata. Results from this study indicate that even though the biology of a copepod might be unaffected by high-CO2, Labidocera spp. are highly vulnerable to ocean acidification.


Assessing conservation opportunity on private land: socio-economic, behavioral, and spatial dimensions.
This study presents a method for assessing conservation opportunity on private land based on landholders' socio-economic, behavioral, and farm characteristics. These characteristics include age, gender, education, level of off-farm income, farm size, proportion of remnant native vegetation on-farm, and ecological value of native vegetation on-farm. A sample of landholders who own greater than 2 ha of land in the South Australian Murray-Darling Basin region were sent a mail-based survey about their values and preferences for environmental management (N = 659, 52% response). Cross-tabulations and ANOVA statistical analysis techniques were used to compare the socio-economic attributes across three landholder classes: disengaged, moderately engaged, and highly engaged in native vegetation planting. Results indicate that highly engaged landholders were more likely to be female, formally educated, hobby farmers who managed small parcels of land and have high off-farm incomes, whereas disengaged landholders held significantly stronger farming connections (more farming experience, family have lived on the farm for more generations). Spatial analysis revealed area-specific differences in conservation opportunity and conservation priority. In some areas, properties of high ecological value were managed by highly engaged landholders, but nearby properties of high value were managed by moderately engaged or disengaged landholders. Environmental managers therefore cannot assume areas of high conservation priority will be areas of high conservation opportunity. At the regional scale, the potential for revegetation seems most promising within the moderately engaged landholder group considering the vast amount of land managed by this group in areas of high ecological value, particularly within the less represented Mallee and Coorong and Rangelands sub-regions. We suggest that incentive schemes which purchase conservation need to be targeted at disengaged landholders; mentoring schemes led by commercial farmers highly engaged in native vegetation planting should be directed at moderately engaged landholders, and; awards programs which acknowledge conservation successes should be targeted at highly engaged landholders.


Feasibility and potential acceptability of three cervical barriers among vulnerable young women in Zimbabwe.
We explored the potential acceptability of three cervical barriers (CB) (Ortho All-Flex diaphragm, SILCS diaphragm, FemCap cervical cap) among sexually experienced Zimbabwean young women. Forty-five young women (aged 16-21 years) received an individual CB educational session. Participants were then randomly assigned to one of the three CBs in a 1:1:1 ratio, and practised insertion and removal of their device at the clinic. Next, participants were interviewed on their practice experiences, and their post-practice attitudes towards CB. All 45 young women were willing and able to insert their assigned device. The majority reported "easy" insertion and removal and 93% "liked" the device they tried. All showed interest in participating in future CB studies: when asked which device they would like to try in the future, over half (58%) chose SILCS, regardless of the device they had tried. The majority felt comfortable touching their genitals to insert/remove the CB and most participants favoured methods' attributes associated with female-control and non-interference with sex. Over half the participants said they would prefer to use a CB continuously compared to episodic use. Two-thirds of them expressed interest in CB for dual protection. The concept of CB, and initial insertion experience, were well accepted in this selected, small group of Zimbabwean young women. Evaluating CB in larger studies seems feasible in this population.


Does caffeine ingestion before a short-term sprint interval training promote body fat loss?
We investigated the effect of caffeine ingestion combined with a 2-wk sprint interval training (SIT) on training-induced reductions in body adiposity. Twenty physically-active men ingested either 5 mg/kg of cellulose as a placebo (PLA, n=10) or 5 mg/kg of caffeine (CAF, n=10) 60 min before each SIT session (13×30 s sprint/15 s of rest). Body mass and skinfold thickness were measured pre- and post-training. Energy expenditure was measured at rest, during exercise, and 45 min after exercise in the first SIT session. Body fat was similar between PLA and CAF groups at pre-training (P>0.05). However, there was a significant decrease in body fat after training in the CAF group (-5.9±4.2%, P<0.05) but not in PLA (1.5±8.0%, P>0.05). There was no difference in energy expenditure at rest and during exercise between PLA and CAF groups (P>0.05), but the post-exercise energy expenditure was 18.3±21.4% greater in the CAF than in the PLA group (P<0.05). In conclusion, caffeine ingestion before SIT sessions induced a body fat loss that may be associated with higher post-exercise energy expenditure.


Eating habits and appetite control: a psychobiological perspective.
An individual's eating behaviour is shaped by factors ranging from economic conditions and cultural practices to biological influences. The physiological system controlling appetite appears to be adapted to solving the problem of an unevenness of food supply across time, and is fairly permissive in its response to undereating and overeating. Consequently, when food is abundant, the diet is energy dense and energy expenditure is low, there is a strong tendency to become obese (i.e. obesity is better viewed as due to a 'toxic' environment than to faulty physiological control of appetite). Under such conditions the most common method of avoiding obesity is through the cognitive control of eating. However, dietary restraint and dieting are demanding tasks, and are associated with psychological costs, including significant impairment of cognitive performance. Restraint is also prone to disinhibition, with the result that it can sometimes undermine eating control, even leading to the development of highly disordered eating patterns. In part, these difficulties are due to the self-perpetuating nature of dietary habits: for example, hunger tends to be diminished during strict unbroken dieting, but increased in individuals having a highly variable eating pattern (such as occurs when eating is frequently disinhibited). These features of appetite control provide both barriers and opportunities for changing behaviour. Accordingly, there is a need for future research to focus on the psycho-social factors and the dieting practices predicting successful eating and weight control, with the objective of identifying the actual cognitive and behavioural strategies used by the many dieters and restrained eaters who are able to achieve weight loss and maintain long-term weight stability.


Exogenous spatial attention: evidence for intact functioning in adults with autism spectrum disorder.
Deficits or atypicalities in attention have been reported in individuals with autism spectrum disorder (ASD), yet no consensus on the nature of these deficits has emerged. We conducted three experiments that paired a peripheral precue with a covert discrimination task, using protocols for which the effects of covert exogenous spatial attention on early vision have been well established in typically developing populations. Experiment 1 assessed changes in contrast sensitivity, using orientation discrimination of a contrast-defined grating; Experiment 2 evaluated the reduction of crowding in the visual periphery, using discrimination of a letter-like figure with flanking stimuli at variable distances; and Experiment 3 assessed improvements in visual search, using discrimination of the same letter-like figure with a variable number of distractor elements. In all three experiments, we found that exogenous attention modulated visual discriminability in a group of high-functioning adults with ASD and that it did so in the same way and to the same extent as in a matched control group. We found no evidence to support the hypothesis that deficits in exogenous spatial attention underlie the emergence of core ASD symptomatology.


Twenty years' experience with myelomeningocele management at a single institution: lessons learned.
The authors reviewed 20 years' experience with the surgical management of open myelomeningocele in a well-defined retrospective cohort from a single large academic medical center. Their goal was to define the characteristics of a modern cohort of children with myelomeningocele to allow for evidence-based decision-making for the treatment of these patients. After IRB approval was obtained, the authors queried an operative database maintained by the Department of Neurological Surgery at Children's Hospital of Pittsburgh for patients who underwent closure of a myelomeningocele between 1995 and 2015. They identified 153 infants, and a retrospective chart review was performed. Eighty-eight percent of the patients required placement of a ventriculoperitoneal shunt, and 15% of these patients acquired shunt-related infections. Eighteen percent of patients underwent Chiari malformation type II (CM-II) decompression. Sixteen percent of patients underwent a tethered cord release. Three percent of patients died within the 1st year of life. Predictors of an early demise included poor Apgar scores, large head circumference, and need for early CM-II decompression. Functional motor outcome was slightly better than predicted by anatomical level of defect. Myelomeningoceles represent a severe birth defect with life-threatening complications. The authors provide long-term follow-up data and insight into factors that contribute to early death.


Comparison of Multivendor Single-Voxel MR Spectroscopy Data Acquired in Healthy Brain at 26 Sites.
Background The hardware and software differences between MR vendors and individual sites influence the quantification of MR spectroscopy data. An analysis of a large data set may help to better understand sources of the total variance in quantified metabolite levels. Purpose To compare multisite quantitative brain MR spectroscopy data acquired in healthy participants at 26 sites by using the vendor-supplied single-voxel point-resolved spectroscopy (PRESS) sequence. Materials and Methods An MR spectroscopy protocol to acquire short-echo-time PRESS data from the midparietal region of the brain was disseminated to 26 research sites operating 3.0-T MR scanners from three different vendors. In this prospective study, healthy participants were scanned between July 2016 and December 2017. Data were analyzed by using software with simulated basis sets customized for each vendor implementation. The proportion of total variance attributed to vendor-, site-, and participant-related effects was estimated by using a linear mixed-effects model. P values were derived through parametric bootstrapping of the linear mixed-effects models (denoted Pboot). Results In total, 296 participants (mean age, 26 years ± 4.6; 155 women and 141 men) were scanned. Good-quality data were recorded from all sites, as evidenced by a consistent linewidth of N-acetylaspartate (range, 4.4-5.0 Hz), signal-to-noise ratio (range, 174-289), and low Cramér-Rao lower bounds (≤5%) for all of the major metabolites. Among the major metabolites, no vendor effects were found for levels of myo-inositol (Pboot > .90), N-acetylaspartate and N-acetylaspartylglutamate (Pboot = .13), or glutamate and glutamine (Pboot = .11). Among the smaller resonances, no vendor effects were found for ascorbate (Pboot = .08), aspartate (Pboot > .90), glutathione (Pboot > .90), or lactate (Pboot = .28). Conclusion Multisite multivendor single-voxel MR spectroscopy studies performed at 3.0 T can yield results that are coherent across vendors, provided that vendor differences in pulse sequence implementation are accounted for in data analysis. However, the site-related effects on variability were more profound and suggest the need for further standardization of spectroscopic protocols. © RSNA, 2020 Online supplemental material is available for this article.


Clinical spectrum of pontine infarction. Clinical-MRI correlations.
We sought clinical and radiological findings of 150 consecutive patients with acute isolated pontine infarct who were admitted to our Stroke Unit over 6 years. In all patients CT, MRI and magnetic resonance angiography (MRA) were performed during the hospitalization. On clinico-radiological analysis regarding the pontine lesion boundaries there were five main clinical patterns that depended on the constant territories of intrinsic pontine arteries: (1). anteromedial pontine syndrome (58%) presented with motor deficit with dysarthria, ataxia, and mild tegmental signs in one third of patients; (2). anterolateral pontine syndrome (17%) developed with motor and sensory deficits in half of the patients, and were associated with tegmental signs (56%) more frequently than the anteromedial infarct syndrome; (3). tegmental pontine syndrome (10%) presented with mild motor deficits and associated with sensory syndromes, eye movement disorders and vestibular system symptoms including vertigo, dizziness and ataxia; (4). bilateral pontine syndrome (11%) consisted with transient consciousness loss, tetraparesis and acute pseudobulbar palsy; (5). unilateral multiple pontine infarcts (4%) were rarely observed, and were always associated with severe sensory-motor deficits and tegmental signs. In our series, there was no infarct in the extreme dorsal and lateral tegmental pontine territories which have been mostly associated with cerebellar infarctions. The main etiology of stroke was basilar artery branch disease (BABD) in 59 patients (39%), followed by small-artery disease (SAD) in 31 (21%), large-artery disease of vertebrobasilar arteries in 27 patients (18%), cardioembolism in 12 (8%) and in 16 patients (11%) no cause of stroke was found. Our findings suggest that it is possible to identify clinical subgroups of pontine infarction, in which BABD and SAD were the most common causes of stroke. After an acute onset, outcome is in general excellent except in those with bilateral pontine lesions.


The liminal space palliative care volunteers occupy and their roles within it: a qualitative study.
Volunteers have an important place in palliative care (PC), positively influencing quality of care for seriously ill people and those close to them and providing a link to the community. However, it is not well understood where volunteers fit into PC provision or how to support them adequately. We therefore chose to describe volunteer roles across care settings through the perspective of those closely involved in the care of terminally ill people. A qualitative study was conducted using both focus groups with volunteers, nurses, psychologists and family physicians and individual semistructured interviews with patients and family caregivers. Participants were recruited from hospital, home, day care and live-in services. 79 people participated in the study. Two volunteer roles were identified. The first was 'being there' for the dying person. Volunteers represent a more approachable face of care, focused on psychological, social and existential care and building relationships. The second was the 'liaison' role. Volunteers occupy a liminal space between the professional and the family domain, through which they notice and communicate patient needs missed by other caregivers. Patient-volunteer matching was a facilitator for role performance; barriers were lack of communication opportunities with professional caregivers and lack of volunteer coordination. Volunteers complement professional caregivers by (1) occupying a unique space between professionals, family and patients and fulfilling a liaison function and (2) being a unique face of care for patients. Healthcare services and policy can support volunteer role performance by ensuring frequent communication opportunities and volunteer coordination.


A placebo-controlled 18-month trial of lamotrigine and lithium maintenance treatment in recently depressed patients with bipolar I disorder.
The anticonvulsant lamotrigine was previously shown to be effective for bipolar depression. This study assessed the efficacy and tolerability of lamotrigine and lithium compared with placebo for the prevention of mood episodes in bipolar disorder. During an 8- to 16-week open-label phase, lamotrigine (titrated to 200 mg/day) was added to current therapy for currently or recently depressed DSM-IV-defined bipolar I outpatients (N = 966) and concomitant drugs were gradually withdrawn. Patients stabilized on open-label treatment (N = 463) were then randomly assigned to lamotrigine (50, 200, or 400 mg/day; N = 221), lithium (0.8-1.1 mEq/L; N = 121), or placebo (N = 121) monotherapy for up to 18 months. The primary outcome measure was time from randomization to intervention (addition of pharmacotherapy) for any mood episode (depressive, manic, hypomanic, or mixed). Data were gathered from September 1997 to August 2001. Time to intervention for any mood episode was statistically superior (p = .029) for both lamotrigine and lithium compared with placebo-median survival times were 200, 170, and 93 days, respectively. Intervention for depression was more frequent than for mania by a factor of nearly 3:1. Lamotrigine was statistically superior to placebo at prolonging the time to intervention for a depressive episode (p = .047). The proportions of patients who were intervention-free for depression at 1 year were lamotrigine 57%, lithium 46%, and placebo 45%. Lithium was statistically superior to placebo at prolonging the time to intervention for a manic or hypomanic episode (p = .026). The proportions of patients who were intervention-free for mania at 1 year were lamotrigine 77%, lithium 86%, and placebo 72%. Headache was the most frequent adverse event for all 3 treatment groups. Lamotrigine and lithium were superior to placebo for the prevention of mood episodes in bipolar I patients, with lamotrigine predominantly effective against depression and lithium predominantly effective against mania.


In vivo study of spherical gold nanoparticles: inflammatory effects and distribution in mice.
Gold nanoparticles (AuNPs) of 21 nm have been previously well characterized in vitro for their capacity to target macrophages via active uptake. However, the short-term impact of such AuNPs on physiological systems, in particular resident macrophages located in fat tissue in vivo, is largely unknown. This project investigated the distribution, organ toxicity and changes in inflammatory cytokines within the adipose tissue after mice were exposed to AuNPs. Male C57BL/6 mice were injected intraperitoneally (IP) with a single dose of AuNPs (7.85 μg AuNPs/g). Body weight and energy intake were recorded daily. Tissues were collected at 1 h, 24 h and 72 h post-injection to test for organ toxicity. AuNP distribution was examined using electron microscopy. Proinflammatory cytokine expression and macrophage number within the abdominal fat pad were determined using real-time PCR. At 72 hours post AuNP injection, daily energy intake and body weight were found to be similar between Control and AuNP treated mice. However, fat mass was significantly smaller in AuNP-treated mice. Following IP injection, AuNPs rapidly accumulated within the abdominal fat tissue and some were seen in the liver. A reduction in TNFα and IL-6 mRNA levels in the fat were observed from 1 h to 72 h post AuNP injection, with no observable changes in macrophage number. There was no detectable toxicity to vital organs (liver and kidney). Our 21 nm spherical AuNPs caused no measurable organ or cell toxicity in mice, but were correlated with significant fat loss and inhibition of inflammatory effects. With the growing incidence of obesity and obesity-related diseases, our findings offer a new avenue for the potential development of gold nanoparticles as a therapeutic agent in the treatment of such disorders.


ORF36 protein kinase of Kaposi's sarcoma herpesvirus activates the c-Jun N-terminal kinase signaling pathway.
alpha-, beta-, and gamma-Herpesviruses encode putative viral protein kinases. The herpes simplex virus UL13, varicella-zoster virus ORF47, and Epstein-Barr virus BGLF4 genes all show protein kinase domains in their protein sequences. Mutational analysis of these herpesviruses demonstrated that the viral kinase is important for optimal virus growth. Previous studies have shown that ORF36 of Kaposi's sarcoma herpesvirus (KSHV) has protein kinase activity and is autophosphorylated on serine. The gene for ORF36 is expressed during lytic growth of the virus and has been classified as a late gene. Inspection of the ORF36 sequence indicated potential motifs that could be involved in activation of cellular transcription factors. To analyze the function of ORF36, the cDNA for this viral gene was tagged with the FLAG epitope and inserted into an expression vector for mammalian cells. Transfection experiments in 293T and SLK cells demonstrated that expression of ORF36 resulted in phosphorylation of the c-Jun N-terminal kinase. Autophosphorylation of ORF36 is important for JNK activation because a mutation in the predicted catalytic domain of ORF36 blocked its ability to phosphorylate JNK. Western blot analysis, using phosphospecific antibodies, revealed that mitogen-activated kinases MKK4 and MKK7 were phosphorylated by ORF36 but not by the kinase-negative mutant. Binding experiments in transfected cells also demonstrated that both the wild type and kinase-negative mutant of ORF36 form a complex with JNK, MKK4, and MKK7. In addition, using a tetracycline-inducible Rta BCBL-1 cell line (TREx BCBL1-Rta), JNK was phosphorylated during lytic replication, and inhibition of JNK activation blocked late viral gene expression but not early viral gene expression. In summary, these studies demonstrate that KSHV ORF36 activates the JNK pathway; thus this cell signaling pathway may function in the KSHV life cycle by regulating viral and/or cellular transcription.


Persistent organic pollutants in liver of Brazilian sharpnose shark (Rhizoprionodon lalandii) from southeastern coast of Brazil.
In the present study, persistent organic pollutants (POPs) were determined in 14 livers from specimens of the Brazilian sharpnose shark (Rhizoprionodon lalandii), which is an important economic resource for small-scale fisheries on the southeastern coast of Brazil. The following concentrations (lipid weight) of POPs were found: ∑PCBs: 1019±267 ng g(-1); ∑DDTs: 111±40 ng g(-1) and ∑PBDEs: 10.4±4.78. PCB 153 made the greatest contribution to ∑PCB (21.4%), followed by PCB 138 (14.6%) and PCB 180 (9.94%). Among chlorinated pesticides, only the p,p'-DDE and p,p'-DDD isomers had concentrations above the detection limit. Moreover, levels above the detection limit were found only for PBDE congeners 47 and 100 (BDEs 47>99). On average, BDE 47 accounted for 88% of the total PBDE load. The feeding habits of the Brazilian sharpnose shark close to the Brazilian coastline are likely the most important difference regarding the accumulation of POPs in comparison to oceanic species that feed in deeper waters. Thus, this species may be used to evaluate the pollution of coastal areas as well as human exposure to contaminants, as the Brazilian sharpnose shark is a frequently used for human consumption. Further studies in other areas of Brazil and involving other species are needed to clarify the mechanisms and potential impact of POPs, which can affect the biology of different organisms and cause population declines.


Isolation of Proteins on Nascent DNA in Hypoxia and Reoxygenation Conditions.
During S-phase both DNA replication and histone deposition must be co-ordinated at and around the replication fork. Replication stress can interfere with the fidelity of this process and can result in genomic instability. The study of proteins associated with DNA replication forks is important for a detailed understanding of DNA replication and chromatin assembly both under basal as well as replication stress conditions. iPOND (isolation of Proteins on Nascent DNA) allows the temporal study of proteins and protein modifications associated with replication forks in a variety of conditions, allowing the 'tracing' of protein association and histone deposition and maturation at active, stalled and damaged replication forks. Importantly, low oxygen (hypoxic) conditions, found in tumours, can result in replication stress. Here we describe the adaptation of the iPOND technique allowing the isolation of proteins and protein modifications specifically with replication forks undergoing hypoxia-induced replication stress. Furthermore, we describe the adaptation of this method for the study of factors associated with replication forks recovering from hypoxia-induced replication stress following periods of reoxygenation. These adaptations are important in order to study proteins associated with replication forks undergoing replication stress in physiologically relevant conditions.


Are digital interventions for smoking cessation in pregnancy effective? A systematic review and meta-analysis.
Smoking in pregnancy remains a global public health issue due to foetal health risks and potential maternal complications. The aims of this systematic review and meta-analysis were to explore: (1) whether digital interventions for pregnancy smoking cessation are effective, (2) the impact of intervention platform on smoking cessation, (3) the associations between specific Behaviour Change Techniques (BCTs) delivered within interventions and smoking cessation and (4) the association between the total number of BCTs delivered and smoking cessation. Systematic searches of 9 databases resulted in the inclusion of 12 published articles (n = 2970). The primary meta-analysis produced a sample-weighted odds ratio (OR) of 1.44 (95% CI 1.04-2.00, p = .03) in favour of digital interventions compared with comparison groups. Computer-based (OR = 3.06, 95% CI 1.28-7.33) and text-message interventions (OR = 1.59, 95% CI 1.07-2.38) were the most effective digital platform. Moderator analyses revealed seven BCTs associated with smoking cessation: information about antecedents; action planning; problem solving; goal setting (behaviour); review behaviour goals; social support (unspecified); and pros and cons. A meta-regression suggested that interventions using larger numbers of BCTs produced the greatest effects. This paper highlights the potential for digital interventions to improve rates of smoking cessation in pregnancy.


Telephone assessment of cognitive function in the late-onset Alzheimer's disease family study.
Administration of cognitive test batteries by telephone has been shown to be a valid and cost-effective means of assessing cognition, but it remains relatively uncommon in epidemiological research. To develop composite cognitive measures and assess how much of the variability in their scores is associated with mode of test administration (ie, in person or by telephone). Cross-sectional cohort study. Late-Onset Alzheimer's Disease Family Study conducted at 18 centers across the United States. A total of 1584 persons, 368 with dementia, from 646 families. Scores on composite measures of memory and cognitive function derived from a battery of 7 performance tests administered in person (69%) or by telephone (31%) by examiners who underwent a structured performance-based training program with annual recertification. Based in part on the results of a factor analysis of the 7 tests, we developed summary measures of working memory, declarative memory, episodic memory, semantic memory, and global cognition. In linear regression analyses, mode of test administration accounted for less than 2% of the variance in the measures. In mixed-effects models, variability in cognitive scores due to center was small relative to variability due to differences between individuals and families. In epidemiologic research on aging and Alzheimer disease, assessment of cognition by telephone has little effect on performance and provides operational flexibility and a means of reducing both costs and missing data.


Age intrinsic loss of telomere protection via TRF1 reduction in endothelial cells.
Aging is a major factor predisposing for multiple diseases. Telomeres at the ends of chromosomes protect the integrity of chromosomal DNA. A specialized six-protein complex termed shelterin protects the telomere from unwanted interaction with DNA damage pathways. The aim of our study was to evaluate the integrity of telomeres and the stability of telomere protection during aging in endothelial cells (EC). We describe that aging EC can be characterized by an increased cell size (40%, p=0.02) and increased expression of PAI 1 (4 fold, p=0.02), MCP1 (10 fold, p=0.001) and GMCSF (15 fold, p=0.004). Telomeric state in aging cells is defined by an increased telomere oxidation (27%, p=0.01), reduced telomere length (62%, p=0.02), and increased DNA damage foci formation (5% in young EC versus 16% in aged EC, p=0.003). This telomeric dysfunction is accompanied by a reduction in the shelterin component TRF1 (33% mRNA, p=0.001; 24% protein, p=0.007). Overexpression of TRF1 in aging EC reduced telomere-associated DNA damage foci to 5% (p=0.02) and reduced expression levels of MCP1 (18% reduction, p=0.008). Aged EC have increased telomere damage and an intrinsic loss of telomere protection. Reestablishing telomere integrity could therefore be a target for rejuvenating endothelial cell function.


A New Class of Bifunctional Major Histocompatibility Class I Antibody Fusion Molecules to Redirect CD8 T Cells.
Bifunctional antibody fusion proteins engaging effector T cells for targeted elimination of tumor cells via CD3 binding have shown efficacy in both preclinical and clinical studies. Different from such a polyclonal T-cell recruitment, an alternative concept is to engage only antigen-specific T-cell subsets. Recruitment of specific subsets of T cells may be as potent but potentially lead to fewer side effects. Tumor-targeted peptide-MHC class I complexes (pMHCI-IgGs) bearing known antigenic peptides complexed with MHC class I molecules mark tumor cells as antigenic and utilize the physiologic way to interact with and activate T-cell receptors. If, for example, virus-specific CD8(+) T cells are addressed, the associated strong antigenicity and tight immune surveillance of the effector cells could lead to efficacious antitumor treatment in various tissues. However, peptide-MHC class I fusions are difficult to express recombinantly, especially when fused to entire antibody molecules. Consequently, current formats are largely limited to small antibody fragment fusions expressed in bacteria followed by refolding or chemical conjugation. Here, we describe a new molecular format bearing a single pMHCI complex per IgG fusion molecule characterized by enhanced stability and expression yields. This molecular format can be expressed in a full immunoglobulin format and can be designed as mono- or bivalent antibody binders. Mol Cancer Ther; 15(9); 2130-42. ©2016 AACR.


Threonine 48 in the BIR domain of survivin is critical to its mitotic and anti-apoptotic activities and can be phosphorylated by CK2 in vitro.
In this study we report that the protein kinase CK2 phosphorylates survivin specifically on threonine 48 (T48) within its BIR domain, and that T48 is critical to both the mitotic and anti-apoptotic roles of survivin. Interestingly, during mitosis T48 mutants localise normally, but are unable to support cell growth when endogenous survivin is removed by siRNA. In addition, while overexpression of survivin normally confers inhibition of TRAIL-mediated apoptosis, this protection is abolished by mutation of T48. Furthermore in interphase cells depletion of endogenous survivin causes redistribution of T48 mutants from the cytoplasm to the nucleus and treatment of cells expressing survivin-GFP with the CK2 inhibitor TBB phenocopies this nuclear redistribution. Finally, we show T48 mutants have increased affinity for borealin, and that this association and cell proliferation can be restored by introduction of a second mutation at T97. To our knowledge these data are the first to identify T48 as a key regulatory site on survivin, and CK2 as a mediator of its mitotic and anti-apoptotic functions.


The role of the dopamine D1 receptor in social cognition: studies using a novel genetic rat model.
Social cognition is an endophenotype that is impaired in schizophrenia and several other (comorbid) psychiatric disorders. One of the modulators of social cognition is dopamine, but its role is not clear. The effects of dopamine are mediated through dopamine receptors, including the dopamine D1 receptor (Drd1). Because current Drd1 receptor agonists are not Drd1 selective, pharmacological tools are not sufficient to delineate the role of the Drd1. Here, we describe a novel rat model with a genetic mutation in Drd1 in which we measured basic behavioural phenotypes and social cognition. The I116S mutation was predicted to render the receptor less stable. In line with this computational prediction, this Drd1 mutation led to a decreased transmembrane insertion of Drd1, whereas Drd1 expression, as measured by Drd1 mRNA levels, remained unaffected. Owing to decreased transmembrane Drd1 insertion, the mutant rats displayed normal basic motoric and neurological parameters, as well as locomotor activity and anxiety-like behaviour. However, measures of social cognition like social interaction, scent marking, pup ultrasonic vocalizations and sociability, were strongly reduced in the mutant rats. This profile of the Drd1 mutant rat offers the field of neuroscience a novel genetic rat model to study a series of psychiatric disorders including schizophrenia, autism, depression, bipolar disorder and drug addiction.


Implant size and fixation mode strongly influence tissue reactions in the CNS.
The function of chronic brain machine interfaces depends on stable electrical contact between neurons and electrodes. A key step in the development of interfaces is therefore to identify implant configurations that minimize adverse long-term tissue reactions. To this end, we here characterized the separate and combined effects of implant size and fixation mode at 6 and 12 weeks post implantation in rat (n = 24) cerebral cortex. Neurons and activated microglia and astrocytes were visualized using NeuN, ED1 and GFAP immunofluorescence microscopy, respectively. The contributions of individual experimental variables to the tissue response were quantified. Implants tethered to the skull caused larger tissue reactions than un-tethered implants. Small diameter (50 µm) implants elicited smaller tissue reactions and resulted in the survival of larger numbers of neurons than did large diameter (200 µm) implants. In addition, tethering resulted in an oval-shaped cavity, with a cross-section area larger than that of the implant itself, and in marked changes in morphology and organization of neurons in the region closest to the tissue interface. Most importantly, for implants that were both large diameter and tethered, glia activation was still ongoing 12 weeks after implantation, as indicated by an increase in GFAP staining between week 6 and 12, while this pattern was not observed for un-tethered, small diameter implants. Our findings therefore clearly indicate that the combined small diameter, un-tethered implants cause the smallest tissue reactions.


Role of Ucp1 enhancer methylation and chromatin remodelling in the control of Ucp1 expression in murine adipose tissue.
Increasing the expression of the brown adipose tissue-specific gene uncoupling protein-1 (Ucp1) is a potential target for treating obesity. We investigated the role of DNA methylation and histone modification in Ucp1 expression in adipose cell lines and ex vivo murine adipose tissues. Methylation state of the Ucp1 enhancer was studied using bisulphite mapping in murine adipose cell lines, and tissue taken from cold-stressed mice, coupled with functional assays of the effects of methylation and demethylation of the Ucp1 promoter on gene expression and nuclear protein binding. We show that demethylation of the Ucp1 promoter by 5-aza-deoxycytidine increases Ucp1 expression while methylation of Ucp1 promoter-reporter constructs decreases expression. Brown adipose tissue-specific Ucp1 expression is associated with decreased CpG dinucleotide methylation of the Ucp1 enhancer. The lowest CpG dinucleotide methylation state was found in two cyclic AMP response elements (CRE3, CRE2) in the Ucp1 promoter and methylation of the CpG in CRE2, but not CRE3 decreased nuclear protein binding. Chromatin immunoprecipitation assays revealed the presence of the silencing DiMethH3K9 modification on the Ucp1 enhancer in white adipose tissue and the appearance of the active TriMethH3K4 mark at the Ucp1 promoter in brown adipose tissue in response to a cold environment. The results demonstrate that CpG dinucleotide methylation of the Ucp1 enhancer exhibits tissue-specific patterns in murine tissue and cell lines and suggest that adipose tissue-specific Ucp1 expression involves demethylation of CpG dinucleotides found in regulatory CREs in the Ucp1 enhancer, as well as modification of histone tails.


Atrial natriuretic peptide-cyclic GMP relationships in normal humans: effects of dietary sodium intake.
1. The present study was designed to investigate the relationships between circulating atrial natriuretic peptide, plasma and urinary cyclic GMP and sodium excretion under basal conditions and in response to changes in dietary sodium intake. 2. Measurements of plasma atrial natriuretic peptide and plasma and urinary (24h collections) cyclic GMP, sodium and creatinine were made in (i) 30 normotensive subjects on their normal sodium intake and (ii) 12 subjects on the 5th day of a low and on the 5th day of a high sodium intake. 3. Plasma cyclic GMP, urinary cyclic GMP and fractional excretion of cyclic GMP in 30 normotensive subjects on their normal sodium intake were (means +/- SEM) 5.4 +/- 0.5 pmol/ml, 434.5 +/- 31.8 pmol/min and 86.9 +/- 8.6%, respectively. There were significant correlations between urinary cyclic GMP and its corresponding filtered load (r = 0.55) and between the renal clearance of cyclic GMP and that of creatinine (r = 0.44), but there were no significant associations between circulating atrial natriuretic peptide and plasma cyclic GMP or the fractional excretion of cyclic GMP or between urinary sodium and the fractional excretion of cyclic GMP. 5. Plasma atrial natriuretic peptide was significantly raised on the 5th day of the high sodium intake compared with the low sodium intake (10.6 +/- 1.6 versus 4.2 +/- 0.9 pg/ml; P < 0.05). Similarly, there were increases in urinary cyclic GMP excretion (692.3 +/- 43.4 versus 427.4 +/- 41.9 pmol/min, P < 0.05), but there were no significant differences in the fractional excretion of cyclic GMP.(ABSTRACT TRUNCATED AT 250 WORDS)


Animal and management factors influencing grower and finisher pig performance and efficiency in European systems: a meta-analysis.
A meta-analysis on the effects of management and animal-based factors on the performance and feed efficiency of growing pigs can provide information on single factor and interaction effects absent in individual studies. This study analysed the effects of such factors on average daily gain (ADG), feed intake (FI) and feed conversion ratio (FCR) of grower and finisher pigs. The multivariate models identified significant effects of: (1) bedding (P<0.01), stage of growth (P<0.001) and the interaction bedding×lysine (P<0.001) on ADG. ADG was higher on straw compared with no bedding (710 v. 605 g/day). (2) FI was significantly affected by stage of growth (P<0.01), bedding (P<0.01), group composition (P<0.05), group size (P<0.01), feed CP content (P<0.01), ambient temperature (P<0.01) and the interaction between floor space and feed energy content (P<0.001). Pigs housed on straw had a lower FI in comparison with those without (1.44 v. 2.04 kg/day); a higher FI was seen for pigs separated by gender in comparison with mixed groups (2.05 v. 1.65 kg/day); FI had a negative linear relationship with group size, the CP content of the feed and ambient temperature. (3) Stage of growth (P<0.001), feed CP (P<0.001) and lysine content (P<0.001), ambient temperature (P<0.001) and feed crude fibre (CF) content (P<0.01) significantly affected FCR; there were no significant interactions between any factors on this trait. There was an improvement in FCR at higher ambient temperatures, increased feed CP and lysine content, but a deterioration of FCR at higher CF contents. For ADG, the interaction of bedding×lysine was caused by pigs housed without bedding (straw) having higher ADG when on a feed lower in lysine, whereas those with bedding had a higher ADG when on a feed higher in lysine. Interaction effects on FI were caused by animals with the least amount of floor space having a higher FI when given a feed with a low metabolisable energy (ME) content, in contrast to all other pigs, which showed a higher FI with increased ME content. The meta-analysis confirmed the significant effect of several well-known factors on the performance and efficiency of grower and finisher pigs, the effects of some less established ones and, importantly, the interactions between such factors.


A mutant of Arabidopsis lacking the triose-phosphate/phosphate translocator reveals metabolic regulation of starch breakdown in the light.
The chloroplast envelope triose-phosphate/phosphate translocator (TPT) is responsible for carbohydrate export during photosynthesis. Using measurements of carbohydrates, partitioning of assimilated 14CO2, photosynthetic gas exchange, and chlorophyll fluorescence, we show that a mutant of Arabidopsis lacking the TPT increases synthesis of starch compared to the wild type, thereby compensating for a deficiency in its ability to export triose-phosphate from the chloroplast. However, during growth under high light, the capacity for starch synthesis becomes limiting so that the chloroplastic phosphate pool is depleted, resulting in a restriction on electron transport, a reduction in the rate of photosynthesis, and slowed plant growth. Under the same conditions but not under low light, we observe release of 14C label from starch, indicating that its synthesis and degradation occur simultaneously in the light. The induction of starch turnover in the mutant specifically under high light conditions leads us to conclude that it is regulated by one or more metabolic signals, which arise as a result of phosphate limitation of photosynthesis.


Haplotypes in the dystrophin DNA segment point to a mosaic origin of modern human diversity.
Although Africa has played a central role in human evolutionary history, certain studies have suggested that not all contemporary human genetic diversity is of recent African origin. We investigated 35 simple polymorphic sites and one T(n) microsatellite in an 8-kb segment of the dystrophin gene. We found 86 haplotypes in 1,343 chromosomes from around the world. Although a classical out-of-Africa topology was observed in trees based on the variant frequencies, the tree of haplotype sequences reveals three lineages accounting for present-day diversity. The proportion of new recombinants and the diversity of the T(n) microsatellite were used to estimate the age of haplotype lineages and the time of colonization events. The lineage that underwent the great expansion originated in Africa prior to the Upper Paleolithic (27,000-56,000 years ago). A second group, of structurally distinct haplotypes that occupy a central position on the tree, has never left Africa. The third lineage is represented by the haplotype that lies closest to the root, is virtually absent in Africa, and appears older than the recent out-of-Africa expansion. We propose that this lineage could have left Africa before the expansion (as early as 160,000 years ago) and admixed, outside of Africa, with the expanding lineage. Contemporary human diversity, although dominated by the recently expanded African lineage, thus represents a mosaic of different contributions.


[Alcohol and work. Consensus Document of the La.R.A. (Workers representing a risk for others) group].
Current Italian legislation obliges employers to prevent workers who are occupationally at risk or who perform jobs that may be hazardous for the safety or health of third parties from consuming alcohol. The LaRA Group undertook to assess whether the law fully safeguards the health and safety of both workers and third parties, without impinging upon the civil rights of workers. A written document expressing agreement was produced following discussions between doctors, lawyers, bioethicists and social partners. There are gaps and inconsistencies in current laws; the differences in local and regional provisions prevent authorities from applying a single strategy at national level. There should be a change in existing rules under which the employer's obligation to enforce the ban on consumption alcohol in the workplace is enacted solely by the "competent" physician whose institutional role is to safeguard and promote health. Some occupational categories that are subject to a ban on alcohol consumption do not currently under-go health surveillance. For example, if road transport drivers are not exposed to a specific occupational risk foreseen under another law, they can be placed under health surveillance only in those regions where the local laws contemplate this type of control. In other cases, the practice of assessing the risk to third parties and providing for compulsory health surveillance in the Risk Assessment Document, is considered by some jurists to be a "consuetudo praeter legem" and therefore acceptable in a field not yet covered by a specific law, but to be "contra legem" or unlawful by other jurists. Moreover, the competent physician who uses a breathanalyser or tests for alcohol addiction faces an ethical dilemma, since by communicating the results to an employer or authorities responsible for the issuing of licenses, he may be violating his professional oath of secrecy. Furthermore, the emphasis placed on testing has induced companies and inspectors to overlook educational and rehabilitation aspects. It is essential to involve general practitioners, educators and specialist services in addressing the problems of alcohol abuse so as to inform/train, recover and rehabilitate. The few studies available indicate that the rules are poorly enforced and that non-compliance may go unobserved. The Group urges all employers to assess the risk for third parties caused by alcohol abuse and to devise a policy on alcohol. Controlling alcohol-related risks in the workplace calls for a better definition of the roles of Vigilance Bod-ies and Company Physicians together with a shift from a reactive to a proactive attitude of all the parties involved.


"Maybe it's still good?" A qualitative study of factors influencing food waste and application of the E.P.A. Food recovery hierarchy in U.S. supermarkets.
About 31% of post-harvest food available for human consumption is lost or wasted annually in the United States. Roughly one third (43 billion lbs.) of food loss occurs in grocery and other retail food stores. Supermarkets engage in food waste reduction, rescue, and recycling strategies, but little is known about frontline workers' and department managers' perspectives on food discards and strategies to limit waste. We aimed to increase understanding of factors influencing grocery retail employees' food waste decision-making at the store level, and of the perspectives of those frontline supermarket workers and managers responsible for food waste prevention and mitigation. We conducted 20 qualitative semi-structured interviews and used thematic analysis to explore how grocery workers and managers view food waste decision drivers, and how these impact the feasibility and effectiveness of waste reduction and food rescue and recycling in their stores. Workers and managers report personalized discard decision-making and confusion between quality and safety indicators. Interviewees described in-store policies, resources and trainings as lacking or inconsistently applied, leading to variability in food waste prevention, rescue, and recycling. Overall, interview participants considered waste reduction strategies that rescue profitable goods more feasible than other food rescue and recycling efforts like donation and composting. Workers' and managers' perceptions of in-store food waste drivers and views on extant food waste prevention and mitigation efforts point to areas for future research and intervention.


Value of mandatory screening studies in emergency department patients cleared for psychiatric admission.
Laboratory and radiographic studies are often required by psychiatric services prior to admitting emergency patients who are otherwise deemed medically stable. Such testing may represent an unnecessary expense that prolongs emergency department stays without significantly improving care. This study determines the prevalence of such testing and how often it leads to changes in care. We prospectively tracked laboratory testing among psychiatric patients presenting to the emergency departments of two academic tertiary care facilities. For each visit we determined whether laboratory or radiographic studies were ordered, and whether the examination was conducted at the request of the emergency physician as part of a medical screening examination or requested by the psychiatry service. We then determined if this testing changed patient disposition. Our study enrolled 598 patients. Of these, emergency physicians ordered testing as a part of medical screening on 155 patients (25.9%). We found the psychiatry service ordered laboratory or radiographic studies for 191 of 434 patients (44.0%) who emergency physicians determined did not require ancillary testing for medical clearance. Of these 191 patients, only one (0.5%; 95% Confidence Interval: 0.01% - 2.9%) had an abnormal result that led to a change in disposition. Total Medicare reimbursement rates for the additional ancillary testing in this study was $37,682. Ancillary testing beyond what is required for medical clearance of psychiatric emergency patients rarely alters care. Policies that require panels of testing prior to psychiatric admission are costly and appear to be unnecessary.


Development and Validation of a Measure of Birth-Related PTSD for Fathers and Birth Partners: The City Birth Trauma Scale (Partner Version).
Research suggests that some fathers and birth partners can experience post-traumatic stress disorder (PTSD) after witnessing a traumatic birth. Birth-related PTSD may impact on many aspects of fathers' and birth partners' life, including relationship breakdown, self-blame and reducing plans for future children. Despite the potential impact on birth partners' lives there is currently no measure of birth-related PTSD validated for use with birth partners. The current study therefore adapted the City Birth Trauma Scale for use with birth partners. The City Birth Trauma Scale (Partner version) is a 29-item questionnaire developed to measure birth-related PTSD according to DSM-5 criteria: stressor criteria (A), symptoms of re-experiencing (B), avoidance (C), negative cognitions and mood (D), and hyperarousal (E), as well as duration of symptoms (F), significant distress or impairment (G), and exclusion criteria or other causes (H). A sample of 301 fathers/birth partners was recruited online and completed measures of birth-related PTSD, bonding, and demographic details. Results showed the City Birth Trauma Scale (Partner version) had good reliability (α = 0.94) and psychometric and construct validity. The fathers/birth partners version has the same two-factor structure as the original scale: (1) general symptoms and (2) birth-related symptoms, which accounted for 51% of the variance. PTSD symptoms were associated with preterm birth and maternal and infant complications. Overall, the City Birth Trauma Scale (Partner version) provides a promising measure of PTSD following childbirth that can be used in research and clinical practice.


Prediction of MHC-peptide binding: a systematic and comprehensive overview.
T cell immune responses are driven by the recognition of peptide antigens (T cell epitopes) that are bound to major histocompatibility complex (MHC) molecules. T cell epitope immunogenicity is thus contingent on several events, including appropriate and effective processing of the peptide from its protein source, stable peptide binding to the MHC molecule, and recognition of the MHC-bound peptide by the T cell receptor. Of these three hallmarks, MHC-peptide binding is the most selective event that determines T cell epitopes. Therefore, prediction of MHC-peptide binding constitutes the principal basis for anticipating potential T cell epitopes. The tremendous relevance of epitope identification in vaccine design and in the monitoring of T cell responses has spurred the development of many computational methods for predicting MHC-peptide binding that improve the efficiency and economics of T cell epitope identification. In this report, we will systematically examine the available methods for predicting MHC-peptide binding and discuss their most relevant advantages and drawbacks.


Trends and outcomes associated with angiotensin-converting enzyme inhibitors.
Limited recent data are available describing the patterns of use of angiotensin converting enzyme inhibitor (ACEI) therapy in patients with acute myocardial infarction (AMI), particularly from the more generalizable population-based setting. The purpose of this study was to examine trends in the receipt of ACEIs and associated short-term outcomes in patients hospitalized with AMI in a large Northeastern community. We conducted a community-wide study of 7991 patients hospitalized with AMI in all metropolitan Worcester, Massachusetts, medical centers during 8 annual periods between 1990 and 2003. Among all patients, 44% received ACEI therapy during their acute hospitalization. There was a marked increase in the use of ACEIs between 1990 (23%) and 2003 (68%), particularly among those who were not on ACEIs before hospitalization. Patients who were previously on ACEIs were more likely to receive this therapy during hospitalization for AMI than were patients who were not previously on this therapy. Patients treated with ACEIs were significantly less likely to die (adjusted odds ratio [OR] 0.33; 95% confidence interval [CI] 0.27-0.41) during hospitalization than were patients who did not receive this therapy, with benefits observed across all subgroups examined. The results of this observational study demonstrate marked increases in the use of ACEIs in patients with AMI in the community setting and demonstrate the benefits to be gained from use of this therapy. Despite these encouraging trends, there remains room for more optimal use of this therapy.


Harmonizing dilemmas. Siblings of children with DAMP and Asperger syndrome's experiences of coping with their life situations.
The aim of this qualitative study was to describe, from their own perspective and experiences, how siblings of children with deficits in attention, motor control and perception (DAMP) and Asperger syndrome cope with their life situations in their families. Fifteen adolescent females 12-18 years old, siblings of boys with DAMP (8 subjects) and Asperger syndrome (7 subjects), were interviewed. The method used in sampling and analysis of interview protocols was the constant comparative method for grounded theory. The inductive categorization of data produced two core concepts, one about the siblings' life situations in DAMP and Asperger syndrome ('dilemma of requirements and concerns') and one about the siblings' coping processes ('harmonizing'). Of the six categories identified, four were categories of the processes of coping ('gaining understanding', 'gaining independence', 'following a bonding responsibility' and 'balancing'). The qualitative differences between coping processes were related to the two categories of context to cope within the experienced dilemma 'requirements' and 'concerns'. The findings contribute to a deeper understanding of the siblings' life situations, and may be important for health personnel in encounter families and for identifying siblings with special needs. The findings may also aid in the development of preventive programs for siblings of children with DAMP and Asperger syndrome.


Benefits of Deep Transcranial Magnetic Stimulation in Alzheimer Disease: Case Series.
Patients diagnosed with Alzheimer disease (AD) show severe cognitive deficits. Decline in memory, language, and executive function have repeatedly been reported. Although AD affects 60% to 80% of demented elderly patients, there is currently no cure and limited treatment alternatives. The aim of the study was to evaluate the feasibility of stimulating prefrontal cortex (PFC) with deep transcranial magnetic stimulation (dTMS) to ameliorate cognitive deficits in patients suffering from AD. Eleven patients (6 males; mean [SD] age, 76 [7] years) in moderate to severe stages of AD received dTMS over the PFC for 20 sessions. Computerized battery (Mindstreams [MS]) and neuropsychological testing (Addenbrooke Cognitive Examination [ACE]) were used to assess cognitive performance before and after treatment. Compared with baseline, 60% of patients performed better on the MS battery and 77% of patients performed better on the ACE testing at the end of dTMS treatment. None of the patients performed worse on both tests at the end of treatment. The DTMS effects on the group mean in ACE and MS approached significance (P = 0.065 and P = 0.086, respectively). A dTMS-induced improvement in the ACE was significant (P = 0.001) on patients in more progressed stage (n = 6). Change in ACE negatively correlated with score at baseline. In sum, the current report of this novel technique indicates that deep stimulation might lead to preservation and even improvement of cognitive functions, at least during the time of treatment. Further examinations should report of long-term effects of this technique.


Brn3a as a marker of retinal ganglion cells: qualitative and quantitative time course studies in naive and optic nerve-injured retinas.
To characterize Brn3a expression in adult albino rat retinal ganglion cells (RGCs) in naïve animals and in animals subjected to complete intraorbital optic nerve transection (IONT) or crush (IONC). Rats were divided into three groups, naïve, IONT, and IONC. Two-, 5-, 9-, or 14-day postlesion (dpl) retinas were examined for immunoreactivity for Brn3a. Before the injury, the RGCs were labeled with Fluorogold (FG; Fluorochrome, Corp. Denver, CO). Brn3a retinal expression was also determined by Western blot analysis. The proportion of RGCs double labeled with Brn3a and FG was determined in radial sections. The temporal course of reduction in Brn3a(+) RGCs and FG(+) RGCs induced by IONC or IONT was assessed by quantifying, in the same wholemounts, the number of surviving FG-labeled RGCs and Brn3a(+)RGCs at the mentioned time points. The total number of FG(+)RGCs was automatically counted in naïve and injured retinas (2 and 5 dpl) or estimated by manual quantification in retinas processed at 9 and 14 dpl. All Brn3a immunopositive RGCs were counted using an automatic routine specifically developed for this purpose. This protocol allowed, as well, the investigation of the spatial distribution of these neurons. Brn3a(+) cells were only present in the ganglion cell layer and showed a spatial distribution comparable to that of FG(+) cells. In the naïve retinal wholemounts the mean (mean +/- SEM; n = 14) total number of FG(+)RGCs and Brn3a(+)RGCs was 80,251 +/- 2,210 and 83,449 +/- 4,541, respectively. Whereas in the radial sections, 92.2% of the FG(+)RGCs were also Brn3a(+), 4.4% of the RGCs were Brn3a(+)FG(-) and 3.4% were FG(+)Brn3a(-). Brn3a expression pattern was maintained in injured RGCs. The temporal course of Brn3a(+)RGC and FG(+)RGC loss induced by IONC or IONT followed a similar trend, but Brn3a(+)RGCs loss was detected earlier than that of FG(+)RGCs. Independent of the marker used to detect the RGCs, it was observed that their loss was quicker and more severe after IONT than after IONC. Brn3a can be used as a reliable, efficient ex vivo marker to identify and quantify RGCs in control and optic nerve-injured retinas.


Comparison of current-use pesticide and other toxicant urinary metabolite levels among pregnant women in the CHAMACOS cohort and NHANES.
We measured 34 metabolites of current-use pesticides and other precursor compounds in urine samples collected twice during pregnancy from 538 women living in the Salinas Valley of California, a highly agricultural area (1999-2001). Precursors of these metabolites included fungicides, carbamate, organochlorine, organophosphorus (OP), and pyrethroid insecticides, and triazine and chloroacetanilide herbicides. We also measured ethylenethiourea, a metabolite of the ethylene-bisdithiocarbamate fungicides. Repeat measurements of the compounds presented here have not been reported in pregnant women previously. To understand the impact of the women's regional environment on these findings, we compared metabolite concentrations from the CHAMACOS (Center for the Health Assessment of Mothers and Children of Salinas) cohort with U.S. national reference data for 342 pregnant women sampled by the National Health and Nutrition Examination Survey (1999-2002). The eight metabolites detected in > 50% of samples [2,4-dichlorophenol (2,4-DCP); 2,5-dichlorophenol (2,5-DCP); 1- and 2-naphthol; ortho-phenylphenol (ORTH); para-nitrophenol (PNP); 2,4,6-trichlorophenol (2,4,6-TCP); and 3,4,6-trichloro-2-pyridinol (TCPy)] may be related to home or agricultural pesticide use in the Salinas Valley, household products, and other sources of chlorinated phenols. More than 78% of women in this study had detectable levels of at least one of the OP pesticide-specific metabolites that we measured, and > 30% had two or more. The 95th percentile values of six of the most commonly detected (> 50%) compounds were significantly higher among the CHAMACOS women after controlling for age, race, socioeconomic status, and smoking [(2,4-DCP; 2,5-DCP; ORTH; PNP; 2,4,6-TCP; and TCPy); quantile regression p < 0.05]. Findings suggest that the CHAMACOS cohort has an additional burden of precursor pesticide exposure compared with the national sample, possibly from living and/or working in an agricultural area.


The diurnal cortisol cycle and cognitive performance in the healthy old.
Associations between cognitive performance and cortisol have variously been reported for measures of both cortisol level and change, and for some domains of cognitive functioning more than others. In this study, associations between cortisol secretion measures and cognitive performance were examined in 50 healthy older people (mean age 74 years; 34 F /16 M). Participants provided 16 accurately timed saliva samples over 2 consecutive days to determine diurnal profiles of cortisol secretion. Overall cognitive performance (OCP) was measured as the principal component of a comprehensive battery of cognitive tests. Across a 30 year age range, there was a strong inverse correlation between age and OCP. Age and poorer OCP were also associated with an attenuated cortisol awakening response (CAR), defined as the rise from 0-30 min after awakening, and a subsequent less steep fall in cortisol level over the rest of the day. Partialling analyses, suggested that the correlation between fall in cortisol over the day and OCP was independent of age. Both older age and less cortisol change were particularly related to poorer performance on tests of declarative memory and executive functioning. Our conclusions are that during the short post-awakening period, an exception exists to the generally pertaining association between higher levels of cortisol and poorer cognitive performance. Consequentially dynamic measures reflecting the rise (CAR) and fall from the post-awakening peak may be particularly salient in helping to explain links between cortisol and cognitive performance. Finally our pattern of results across different cognitive tests suggests an association between cortisol and those domains of cognitive functioning which depend crucially on the integrity of the hippocampus and pre-frontal cortex.


Medicare Program: Hospital Outpatient Prospective Payment and Ambulatory Surgical Center Payment Systems and Quality Reporting Programs. Final rule with comment period.
This final rule with comment period revises the Medicare hospital outpatient prospective payment system (OPPS) and the Medicare ambulatory surgical center (ASC) payment system for CY 2018 to implement changes arising from our continuing experience with these systems. In this final rule with comment period, we describe the changes to the amounts and factors used to determine the payment rates for Medicare services paid under the OPPS and those paid under the ASC payment system. In addition, this final rule with comment period updates and refines the requirements for the Hospital Outpatient Quality Reporting (OQR) Program and the ASC Quality Reporting (ASCQR) Program.


Health behaviors and mental health during the COVID-19 pandemic: A longitudinal population-based survey in Germany.
To understand the mental health response to repeated and prolonged stress during the COVID-19 related lockdown and the role of specific health behaviors to buffer against this stress. In a longitudinal study with several measurement points covering three months during the COVID-19 pandemic, about 3500 randomly selected participants representative of the German population reported on their mental health (anxiety, depression, loneliness) and health behaviors (screen time, snack consumption, physical activity). Symptoms of anxiety, depression, and loneliness were highest shortly after the lockdown came into effect. Over time, the symptoms were stable or went down slightly, corresponding to patterns of habituation. Among people with higher vulnerability to poor mental health during the lockdown (e.g., women), the proportion with high levels of anxiety, depression, and loneliness was considerably larger. These groups also reported fewer health-promoting behaviors. More screen time, more snacking, and less physical activity were related to higher symptoms of anxiety, depression, and loneliness across all time points. Changes in health behaviors over time mostly did not predict changes in mental health symptoms. Mental health and engagement in protective health behaviors was lowest at the start of the lockdown. Health behaviors mostly returned to pre-lockdown levels within three months. Engaging in healthier behaviors was associated with better mental health. Policy implications of these findings are discussed. This study provides important insights into (unintended) side effects of an international crisis and can contribute to a better understanding of how to preserve mental health.


Differential expression of plasminogen activator inhibitor-1, tumor necrosis factor-alpha, TNF-alpha converting enzyme and ADAMTS family members in murine fat territories.
Our objective was to investigate expression of A disintegrin and metalloproteinase (ADAM) and ADAM proteins with a thrombospondin (TS) motif (ADAMTS) family members in adipose tissue of lean and obese mice. Five-week-old male mice were kept on standard chow (SFD) or on high fat diet (HFD) for 15 weeks, and subcutaneous (SC) and gonadal (GON) adipose tissue, as well as mature adipocytes and stromal-vascular (S-V) cells were harvested. mRNA levels of plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor-alpha (TNF-alpha), ADAM-17 (TACE or TNF-alpha converting enzyme), ADAMTS-1 and ADAMTS-8 were quantified in isolated adipose tissues and cell fractions, and during differentiation of murine preadipocytes. The HFD resulted in a significantly enhanced weight of isolated SC and GON fat pads, and in enhanced blood levels of glucose, cholesterol and PAI-1. ADAM-17, TNF-alpha, PAI-1, ADAMTS-1 and ADAMTS-8 mRNA were detected in both SC and GON adipose tissue of lean mice (SFD). In SC adipose tissue of obese mice (HFD), the expression of ADAM-17 and PAI-1 was enhanced and that of ADAMTS-1 reduced, whereas in GON adipose tissue expression of TNF-alpha was enhanced and that of ADAMTS-8 reduced. In lean and obese mice, expression of ADAM-17, ADAMTS-1 and ADAMTS-8 was higher in the S-V cell fraction than in mature adipocytes. During differentiation of murine 3T3-F442A preadipocytes, expression of ADAM-17 and ADAMTS-1 remained virtually unaltered, whereas that of ADAMTS-8 decreased as adipocytes matured. Several ADAM and ADAMTS family members are expressed in adipose tissue and during differentiation of preadipocytes. Modulation of their expression upon development of obesity is adipose tissue-dependent.


Tissue specificity of tropomyosin from the crayfish, Cambarus clarki.
The molecular heterogeneity and tissue specificity of crustacean tropomyosin were investigated, using muscle and nonmuscle tissues from the crayfish, Cambarus clarki. In muscle, three types of tropomyosin isoforms were found on two-dimensional gel electrophoresis. One of them was specific to cardiac muscle, and the other two were shared by skeletal and visceral muscles. In nonmuscle tissues, four types of isoforms were found on two-dimensional gel electrophoresis and in immunoreplica tests using an antiserum against crayfish skeletal muscle tropomyosin. Two of them were common to the muscle isoforms, but the other two were not detected in muscles. Furthermore, nonmuscle tissues contained several peculiar isoforms, the electrophoretic mobilities of which were considerably higher than those of the other isoforms mentioned above. When tropomyosin was purified from the mid-gut gland, these isoforms with high mobilities were found in the crude tropomyosin preparation. These results showed that the crayfish tropomyosin was heterogeneous and that the isoforms were distributed in a tissue-specific manner, like vertebrate tropomyosin. However, the results did not coincide with those of our previous study on horseshoe crab tropomyosin, which showed molecular heterogeneity but no tissue specificity. In view of the difference in the isoform distributions between the two major groups (Crustacea and Merostomata) of Arthropoda, the significance of the tissue specificity of tropomyosin isoforms was discussed.


An examination of interprofessional education in a pre-registration children's nursing course.
Aim This study set out to gain insight into views held by children's nursing students, lecturers and clinically based nurse mentors about interprofessional education (IPE). Method A qualitative research design was undertaken. Small focus groups were held with pre-registration children's nursing students, and semi-structured interviews with university lecturers and clinically based nurse mentors to find out information about their understanding of IPE. Findings Participants not only valued IPE, but they agreed that it had the potential to positively affect the care of children, young people and their families. Conclusion It is important that professionals collaborate to ensure students receive high quality IPE learning experiences.


The effects of the 1997 Balanced Budget Act on family practice residency training programs.
This study assessed the impact of the Balanced Budget Act (BBA) of 1997 on family practice residency training programs in the United States. We surveyed 453 active family practice residency programs, asking about program closures and new program starts (including rural training tracks), changes in the number of residents and faculty, and curriculum changes. Programs were classified according to their urban or rural location, university or community hospital setting, and rural and/or urban underserved mission emphasis. A total of 435 (96%) of the programs responded. Overall, the impact of the BBA was relatively small. In 1998 and 1999, nationwide, there were 11 program closures, a net decrease of only 82 residents, and a net increase of 52 faculty across program settings and mission emphasis. The rate of family practice residency program closures increased from an average of 3.0 per year between 1988-1997 to 4.8 per year in the 4 years following passage of the BBA. The 1997 BBA did not have an immediate significant negative impact on family practice residency programs. However, there is a worrisome increase in the rate of family practice residency closures since 1997. A mechanism needs to be established to monitor all primary care program closures to give an early warning should this trend continue.


Population-based study of malignancies and HIV infection among injecting drug users in a New York City methadone treatment program, 1985-1991.
To study the incidence of AIDS-defining and non-AIDS-defining malignancies in injecting drug users with and without HIV infection in a methadone maintenance treatment program (MMTP). Prospective study within a hospital-affiliated MMTP with on-site primary medical services. The MMTP has been the site of a voluntary longitudinal cohort study of HIV infection since 1985. Active surveillance for all new cancer cases occurring among patients in the MMTP between July 1985 and August 1991. Cancer cases were identified by review of clinic and hospital records, hospital-based tumor registries, and New York City vital records. Cancer incidence was determined for the overall MMTP population and for HIV-seropositive and HIV-seronegative cohort study subgroups. During the study period the MMTP population comprised 2174 patients followed for 5491 person-years; 844 patients (380 HIV-seropositive, 464 HIV-seronegative) also participated in the cohort study. Fifteen non-AIDS-defining malignancies occurred among all MMTP patients (2.73 cases per 1000 person-years); the most frequent sites were lung, larynx, and cervix (n = 6, 2 and 2, respectively). Eighty per cent of patients with these cancer diagnoses and known HIV serologic status were seropositive. Within the cohort study group, six out of 380 HIV-seropositives developed non-AIDS-defining cancers versus one out of 464 HIV-seronegatives (P = 0.05, Fisher's exact test). Lung cancer cases in HIV-seropositive patients tended to occur at an earlier age and was more aggressive than in patients with HIV-seronegative or unknown status. During the same period, two cases of AIDS-defining lymphoma and one case of Kaposi's sarcoma were diagnosed in the MMTP population (0.5 cases per 1000 person-years). Solid neoplasms, while infrequent, were associated with HIV infection and were more common than AIDS-defining cancers in this population of drug injectors. Further study is needed to explore the relationship between HIV, behavioral factors, and cancer risk in injecting drug users.


Effect of the intermittent atmosphere on laser scintillations.
The evolution of the scintillation index of a spherical wave propagating through randomly varying profiles of the strength and the smallest scale of turbulence is calculated. These parameters were taken to be jointly log normal along the range. The specific parameters of the probability-density function that were used were taken from stratospheric experiments conducted by the U.S. Air Force.


Physical symptoms and physician-experienced difficulty in the physician-patient relationship.
The practice of medicine has many expected and accepted challenges, but all physicians experience some patients as difficult to a degree that transcends these expectations. Physician-experienced difficulty is associated with a syndrome of three characteristics: patient psychopathology, abrasive interpersonal styles, and multiple physical symptoms. To assess the roles played by the number of physical symptoms and by specific symptoms in determining whether physician-experienced difficulty occurs. New analyses of epidemiologic survey data from the Primary Care Evaluation of Mental Disorders (PRIME-MD) 1000 Study. Four primary care clinics. 627 ambulatory patients seen by 27 physicians. Physician-experienced difficulty was measured by using the 10-item Difficult Doctor Patient Relationship Questionnaire (DDPRQ-10); patient-reported physical symptoms and physician-assessed psychopathology and somatoform symptoms were evaluated by using the PRIME-MD; and physical illnesses were measured by using a physician questionnaire. The number of physical symptoms and the number of somatoform symptoms correlated with difficulty (r = 0.39 and r = 0.37, respectively; P < 0.001), and the correlations remained significant after adjustment for physical and mental disorders (r = 0.20 for both correlations; P < 0.001). Difficult patients were more likely to have each of 16 physical symptoms; the odds of being difficult were greater for patients with 1 of 5 particular symptoms (stomach pain, fainting, loose stools/diarrhea, palpitations, and sleep problems), even after adjustment for physical and mental disorders. All 10 items on the DDPRQ-10 were influenced by physical symptoms, particularly those items that asked about physician frustration and whether patients were manipulative and time consuming. The association between physical symptoms and difficulty is due in part to the association between physical symptoms and mental disorders, but symptoms also contribute independently to difficulty. The independent component of symptom-associated difficulty may be due to 1) differences between patient and physician in expectations about treatment and 2) the part that symptoms play in conferring the "sick role" on a patient.


Neuronal correlates of altered empathy and social cognition in borderline personality disorder.
Borderline personality disorder (BPD) is a severe psychiatric condition of undetermined brain underpinnings, which involves profound emotion regulation deficits and interpersonal impairment. To elucidate biopsychological markers of the disorder, we performed two studies: i.) assessing empathy and social cognition and ii.) measuring the psychophysical properties and functional brain correlates of empathic functioning in a total of fifty-one affected patients and 50 age- and gender-matched controls. In the behavioral study we applied the Multifaceted Empathy Test (MET), a new, ecologically valid measure to assess cognitive (i.e., social cognition) and emotional (i.e., empathic concern) empathy to a subset of participants. In the second study, functional Magnetic Resonance Imaging and skin conductance measurements were performed while participants took a scanner-adapted version of the MET. Patients with BPD showed impairments in cognitive and emotional empathy. Brain responses during cognitive empathy were significantly reduced in patients compared to controls in the left superior temporal sulcus and gyrus (STS/STG), where this reduction was associated with levels of intrusive symptomatology in the BPD group. During emotional empathy, patients with BPD exhibited greater brain activity than controls in the right middle insular cortex, a response that was associated with skin conductance responses in the patients. Results indicate that altered functioning of the STS/STG and insula represents pathophysiological mediators for reduced empathy in BPD, with an important role for intrusive symptomatology and levels of arousal. The findings thus support a conceptualization of BPD as involving deficits in both inferring others' mental states and being emotionally attuned to another person.


THE EFFECTS OF VARIED TENSIONS OF KINESIOLOGY TAPING ON QUADRICEPS STRENGTH AND LOWER LIMB FUNCTION.
Kinesiology Taping (KT) may promote changes in muscle strength and motor performance, topics of great interest in the sports-medicine sciences. These characteristics are purported to be associated with the tension generated by the KT on the skin. However, the most suitable tension for the attainment of these strength and performance effects has not yet been confirmed. The purpose of the present study was to analyze the effects of different tensions of KT on the isometric contraction of the quadriceps and lower limb function of healthy individuals over a period of seven days. Blind, randomized, clinical trial. One hundred and thirty healthy individuals were distributed into the following five groups: control (without KT); KT0 (KT without tension); KT50; KT75 and KT100 (approximately 50%, 75% and 100% tension applied to the tape, respectively). Assessments of isometric quadriceps strength were conducted using a hand held dynamometer. Lower limb function was assessed through Single Hop Test for Distance, with five measurement periods: baseline; immediately after KT application; three days after KT; five days after KT; and 72h after KT removal (follow-up). There were no statistically significant differences (p > 0.05) at any of the studied periods on participants' quadriceps strength nor in the function of the lower dominant limb, based on comparisons between the control group and the experimental groups. KT applied with different tensions did not produce modulations, in short or long-term, on quadriceps' strength or lower limb function of healthy individuals. Therefore, this type of KT application, when seeking these objectives, should be reconsidered. 1b.


International Cooperation in Space: A wide variety of joint projects contribute to science and have practical applications.
NASA has developed an extensive program of international cooperation, which opens the entire range of its space activities to foreign participation and benefit. The program has involved developing as well as advanced countries in activities that are fruitful scientifically, technically, economically, and (it may be hoped) politically. "Gaps" or limitations in the program are found in the areas of manned space flight and large booster development and in cooperation with the Soviet Union. In the first case, the low level of European budgeting is the prime explanation, perhaps reflecting Europe's late start in the space business and an early underestimate of its values and possibilities. In the second case, the explanation seems to rest with Soviet political views rather than with technical problems or any lack of interest by the United States. Openhanded efforts are currently under way to raise the level of foreign participation in the space programs of the next decade, but it is too early to estimate their chances of major success. Public commentary could undoubtedly assist these efforts more effectively if it were directed more clearly to the actual sources of restriction on foreign participation. Despite the restricting factors abroad, however, there is every reason to persevere with existing and improved programs for international cooperation.


To tap or not to tap: high likelihood of meningitis without sepsis among very low birth weight infants.
Neonatal meningitis is associated with significant morbidity and mortality. We speculated that meningitis may be underdiagnosed among very low birth weight (VLBW) infants because of the failure to perform lumbar punctures (LPs) in infants with suspected sepsis. This study was undertaken to review the epidemiology of late-onset meningitis in VLBW (401-1500 g) infants and to evaluate the concordance of cerebrospinal fluid (CSF) and blood culture (BC) results. VLBW infants (excluding those with intraventricular shunts) born at centers of the National Institute of Child Health and Human Development Neonatal Research Network from September 1, 1998, through December 31, 2001, were studied. Late-onset meningitis was defined by culture-based criteria and classified as meningitis with or without associated sepsis. Unadjusted comparisons were made using chi2 tests and adjusted comparisons using regression models. Of 9641 VLBW infants who survived >3 days, 2877 (30%) had > or = 1 LPs, and 6056 (63%) had > or = 1 BC performed after day 3. One hundred thirty-four infants had late-onset meningitis (1.4% of all patients; 5% of those with an LP). Pathogens associated with meningitis were similar to those associated with sepsis. One third (45 of 134) of the infants with meningitis had negative BCs. Lower gestational age and prior sepsis increased risk for meningitis. Compared with uninfected infants, those with meningitis had a longer time on mechanical ventilation (28 vs 18 days), had longer hospitalizations (91 vs 79 days), were more likely to have seizures (25% vs 2%), and were more likely to die (23% vs 2%). Meningitis is a serious complication among VLBW infants, associated with increased severity of illness and risk of death. Of note, one third of the infants with meningitis had meningitis in the absence of sepsis. Because CSF cultures were performed only half as often as BCs, this discordance in blood and CSF culture results suggests that meningitis may be underdiagnosed among VLBW infants.


Kinase cascades regulating entry into apoptosis.
All cells are constantly exposed to conflicting environment cues that signal cell survival or cell death. Survival signals are delivered by autocrine or paracrine factors that actively suppress a default death pathway. In addition to survival factor withdrawal, cell death can be triggered by environmental stresses such as heat, UV light, and hyperosmolarity or by dedicated death receptors (e.g., FAS/APO-1 and tumor necrosis factor [TNF] receptors) that are counterparts of growth factor or survival receptors at the cell surface. One of the ways that cells integrate conflicting exogenous stimuli is by phosphorylation (or dephosphorylation) of cellular constituents by interacting cascades of serine/threonine and tyrosine protein kinases (and phosphatases). Survival factors (e.g., growth factors and mitogens) activate receptor tyrosine kinases and selected mitogen-activated, cyclin-dependent, lipid-activated, nucleic acid-dependent, and cyclic AMP-dependent kinases to promote cell survival and proliferation, whereas environmental stress (or death factors such as FAS/APO-1 ligand and TNF-alpha) activates different members of these kinase families to inhibit cell growth and, under some circumstances, promote apoptotic cell death. Because individual kinase cascades can interact with one another, they are able to integrate conflicting exogenous stimuli and provide a link between cell surface receptors and the biochemical pathways leading to cell proliferation or cell death.


Influence of central obesity in estimating maximal oxygen uptake.
To assess the influence of central obesity on the magnitude of the error of estimate of maximal oxygen uptake in maximal cycling exercise testing. A total of 1,715 adults (68% men) between 18-91 years of age underwent cardiopulmonary exercise testing using a progressive protocol to volitional fatigue. Subjects were stratified by central obesity into three quartile ranges: Q1, Q2-3 and Q4. Maximal oxygen uptake [mL.(kg.min)-1] was estimated by the attained maximal workload and body weight using gender- and population-specific equations. The error of estimate [mL.(kg.min)-1] and percent error between measured and estimated maximal oxygen uptake values were compared among obesity quartile ranges. The error of estimate and percent error differed (mean ± SD) for men (Q1=1.3±3.7 and 2.0±10.4; Q2-3=0.5±3.1 and -0.5±13.0; and Q4=-0.3±2.8 and -4.5±15.8 (p<0.05)) and for women (Q1=1.6±3.3 and 3.6±10.2; Q2-3=0.4±2.7 and -0.4±11.8; and Q4=-0.9±2.3 and -10.0±22.7 (p<0.05)). Central obesity directly influences the magnitude of the error of estimate of maximal oxygen uptake and should be considered when direct expired gas analysis is unavailable.


Lack of concordance between empirical scores and physician assessments of stroke and bleeding risk in atrial fibrillation: results from the Outcomes Registry for Better Informed Treatment of Atrial Fibrillation (ORBIT-AF) registry.
Physicians treating patients with atrial fibrillation (AF) must weigh the benefits of anticoagulation in preventing stroke versus the risk of bleeding. Although empirical models have been developed to predict such risks, the degree to which these coincide with clinicians' estimates is unclear. We examined 10 094 AF patients enrolled in the Outcomes Registry for Better Informed Treatment of AF (ORBIT-AF) registry between June 2010 and August 2011. Empirical stroke and bleeding risks were assessed by using the congestive heart failure, hypertension, age ≥75 years, diabetes mellitus, and previous stroke or transient ischemic attack (CHADS2) and Anticoagulation and Risk Factors in Atrial Fibrillation (ATRIA) scores, respectively. Separately, physicians were asked to categorize their patients' stroke and bleeding risks: low risk (<3%); intermediate risk (3%-6%); and high risk (>6%). Overall, 72% (n=7251) in ORBIT-AF had high-risk CHADS2 scores (≥2). However, only 16% were assessed as high stroke risk by physicians. Although 17% (n=1749) had high ATRIA bleeding risk (score ≥5), only 7% (n=719) were considered so by physicians. The associations between empirical and physician-estimated stroke and bleeding risks were low (weighted Kappa 0.1 and 0.11, respectively). Physicians weighed hypertension, heart failure, and diabetes mellitus less significantly than empirical models in estimating stroke risk; physicians weighted anemia and dialysis less significantly than empirical models when estimating bleeding risks. Anticoagulation use was highest among patients with high stroke risk, assessed by either empirical model or physician estimates. In contrast, physician and empirical estimates of bleeding had limited impact on treatment choice. There is little agreement between provider-assessed risk and empirical scores in AF. These differences may explain, in part, the current divergence of anticoagulation treatment decisions from guideline recommendations. http://www.clinicaltrials.gov. Unique identifier: NCT01165710.


Effect of Antioxidant Addition on Milk Beverage Supplemented with Coffee and Shelf-life Prediction.
This study aimed to extend the shelf-life of coffee-containing milk beverage by adding Theobroma cacao (cacao nibs) extract. To prepare the beverage sample containing cacao nibs extract, 0.8% cacao nibs hydrothermal extract was aseptically injected. Qualitative changes in the beverage samples, including antioxidant effect, peroxide value (POV), caffeine content, and sensory parameters were monitored regularly during storage at 10°C, 20°C, and 30°C for 4 wk. The inclusion of cacao nibs extract produced higher antioxidant activity compared to the control. As the storage temperature increased, the POV of all samples increased. Samples with cacao nibs extract generally displayed lower POV than the control. The caffeine content of all samples tended to decrease during storage, with the decrease accentuated by higher storage temperatures. In the shelf-life prediction using the Arrhenius model, the kinetic regressions of the cacao nibs extract-added sample and control were Y POV=1.2212X-2.1141 (r2=0.9713) and Y POV=1.8075X-2.0189 (r2=0.9883), respectively. Finally, the predicted shelf-life of cacao nibs-added group and control to reach the quality limit (20 meq/kg POV) were approximately 18.11 and 12.18 wk, respectively. The results collectively indicate that the addition of cacao nibs extract extends the shelf-life of the coffee-containing milk beverage and heightened the antioxidant effect.


DNA adduct bypass polymerization by Sulfolobus solfataricus DNA polymerase Dpo4: analysis and crystal structures of multiple base pair substitution and frameshift products with the adduct 1,N2-ethenoguanine.
1,N(2)-Etheno(epsilon)guanine is a mutagenic DNA lesion derived from lipid oxidation products and also from some chemical carcinogens. Gel electrophoretic analysis of the products of primer extension by Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) indicated preferential incorporation of A opposite 3'-(1,N(2)-epsilon-G)TACT-5', among the four dNTPs tested individually. With the template 3'-(1,N(2)-epsilon-G)CACT-5', both G and A were incorporated. When primer extension was done in the presence of a mixture of all four dNTPs, high pressure liquid chromatography-mass spectrometry analysis of the products indicated that (opposite 3'-(1,N(2)-epsilon-G)CACT-5') the major product was 5'-GTGA-3' and the minor product was 5'-AGTGA-3'. With the template 3'-(1,N(2)-epsilon-G)TACT-5', the following four products were identified by high pressure liquid chromatography-mass spectrometry: 5'-AATGA-3', 5'-ATTGA-3', 5'-ATGA-3', and 5'-TGA-3'. An x-ray crystal structure of Dpo4 was solved (2.1 A) with a primer-template and A placed in the primer to be opposite the 1,N(2)-epsilon-G in the template 3'-(1,N(2)-epsilon-G)TACT 5'. The added A in the primer was paired across the template T with classic Watson-Crick geometry. Similar structures were observed in a ternary Dpo4-DNA-dATP complex and a ternary Dpo4-DNA-ddATP complex, with d(d)ATP opposite the template T. A similar structure was observed with a ddGTP adjacent to the primer and opposite the C next to 1,N(2)-epsilon-G in 3'-(1,N(2)-epsilon-G)CACT-5'. We concluded that Dpo4 uses several mechanisms, including A incorporation opposite 1,N(2)-epsilon-G and also a variation of dNTP-stabilized misalignment, to generate both base pair and frameshift mutations.