Activation of the Listeria monocytogenes Virulence Program by a Reducing Environment.

Upon entry into the host cell cytosol, the facultative intracellular pathogen Listeria monocytogenes coordinates the expression of numerous essential virulence factors by allosteric binding of glutathione (GSH) to the Crp-Fnr family transcriptional regulator PrfA. Here, we report that robust virulence gene expression can be recapitulated by growing bacteria in a synthetic medium containing GSH or other chemical reducing agents. Bacteria grown under these conditions were 45-fold more virulent in an acute murine infection model and conferred greater immunity to a subsequent lethal challenge than bacteria grown in conventional media. During cultivation in vitro, PrfA activation was completely dependent on the intracellular levels of GSH, as a glutathione synthase mutant (ΔgshF) was activated by exogenous GSH but not reducing agents. PrfA activation was repressed in a synthetic medium supplemented with oligopeptides, but the repression was relieved by stimulation of the stringent response. These data suggest that cytosolic L. monocytogenes interprets a combination of metabolic and redox cues as a signal to initiate robust virulence gene expression in vivoIMPORTANCE Intracellular pathogens are responsible for much of the worldwide morbidity and mortality from infectious diseases. These pathogens have evolved various strategies to proliferate within individual cells of the host and avoid the host immune response. Through cellular invasion or the use of specialized secretion machinery, all intracellular pathogens must access the host cell cytosol to establish their replicative niches. Determining how these pathogens sense and respond to the intracellular compartment to establish a successful infection is critical to our basic understanding of the pathogenesis of each organism and for the rational design of therapeutic interventions. Listeria monocytogenes is a model intracellular pathogen with robust in vitro and in vivo infection models. Studies of the host-sensing and downstream signaling mechanisms evolved by L. monocytogenes often describe themes of pathogenesis that are broadly applicable to less tractable pathogens. Here, we describe how bacteria use external redox states as a cue to activate virulence

Major coronary artery anomalies in a pediatric population: incidence and clinical importance

AbstractOBJECTIVESWe sought to prospectively determine the incidence and clinical significance of major coronary artery anomalies in asymptomatic children using transthoracic two-dimensional echocardiography.BACKGROUNDAnomalous origins of the left main coronary artery (ALMCA) from the right sinus of Valsalva or anomalous origins the right coronary artery (ARCA) from the left sinus are rarely diagnosed in children and can cause sudden death, especially in young athletes. Because most patients are asymptomatic, the diagnosis is often made post mortem. No study to date has prospectively identified anomalous coronary arteries in asymptomatic children in the general population.METHODSAfter serendipitously identifying an index case with ALMCA, we examined proximal coronary artery anatomy in children with otherwise anatomically normal hearts who were referred for echocardiography. In those diagnosed with ALMCA or ARCA, we performed further tests.RESULTSWithin a three-year period, echocardiograms were obtained in 2,388 children and adolescents. Four children (0.17%) were identified with anomalous origin of their coronary arteries, and angiograms, exercise perfusion studies and/or stress tests were then performed. One ARCA patient had decreased perfusion in the right coronary artery (RCA) perfusion area and showed ventricular ectopy on electrocardiogram (ECG) at rest that diminished but did not resolve with exercise. A second patient with ALMCA had atrial tachycardia immediately after exercise, with inferior and lateral ischemic changes on ECG and frequent junctional and/or ventricular premature complexes both at rest and recovery.CONCLUSIONSThis study demonstrates that although anomalous origins of coronary arteries are rare in asymptomatic children, the prevalence is greater than that found in other prospective studies. Ischemia can occur with both ALMCA and ARCA even though patients remain asymptomatic. Because of the high risk of sudden cardiac death, aggressive surgical management and close follow-up are necessary

Relational Theory and Intergenerational Connectedness: A Qualitative Study

Relational theory encourages women to be connected in relationships. The authors used qualitative methodology to explore the interactions of a nonfamilial intergenerational group of 7 female adolescents (13-15 years) and 5 older women (62-80 years) in a structured retreat. Findings indicated that the participants experienced increased connectivity and feminine identity. © 2010 by the American Counseling Association. All rights reserved

Spectrum of an oscillator with jumping frequency and the interference of partial susceptibilities

We study an underdamped oscillator with shot-noise frequency fluctuations. The oscillator spectrum is determined by the interference of the susceptibilities for different eigenfrequencies. Depending on the parameters, it has a fine structure or displays a single asymmetric peak. For nano-mechanical resonators with a fluctuating number of attached molecules, the spectrum is found in a simple analytical form. The results bear on various types of systems where the reciprocal correlation time of frequency fluctuations can be comparable to the typical frequency jumps

Long-term safety of ospemifene (52-week extension) in the treatment of vulvar and vaginal atrophy in hysterectomized postmenopausal women

Objective To examine the long-term safety of oral ospemifene, a non-estrogen tissue-selective estrogen agonist/antagonist, for the treatment of moderate to severe dyspareunia, a symptom of vulvar and vaginal atrophy (VVA) due to menopause. Study design This multicenter, long-term, open-label, safety extension study was conducted in women without a uterus aged 40–80 years (N = 301) who received oral ospemifene 60 mg/day for 52 weeks. Participants either continued their 60-mg/day ospemifene dose from the initial 12-week pivotal efficacy study or switched from blinded placebo or ospemifene 30 mg/day to open-label ospemifene 60 mg/day. The 52-week open-label extension period plus initial 12-week treatment period totaled up to 64 weeks of ospemifene exposure. A 4-week posttreatment follow-up ensued (68 weeks total). Main outcome measures Safety assessments included adverse events, laboratory studies, physical and gynecologic examination, vital signs, breast palpation, and mammography. Results Most treatment-emergent adverse events (TEAEs) during the extension study were mild or moderate in severity. The most common TEAE related to study drug was hot flushes (10%; leading to discontinuation for 2% of patients). One serious TEAE, a non-ST-elevation myocardial infarction in a patient with pre-existing cardiac disease, was considered possibly related to study medication. One mild breast-related TEAE, considered unrelated to study drug, was ongoing at study completion. There were no instances of pelvic organ prolapse, incontinence, venous thromboembolism, fractures, breast cancers or death. No clinically significant adverse changes were observed in other safety parameters. Conclusions Ospemifene is clinically safe and generally well tolerated in postmenopausal patients with dyspareunia, a symptom of VVA

Does malnutrition influence outcome in children undergoing congenital heart surgery in a developing country?

Background Most children undergoing cardiac surgery for congenital heart disease (CHD) in developing countries are malnourished. Malnutrition is known as a co-morbidity factor that might predict and influence outcomes after surgery. Objectives To evaluate the effect of malnutrition and other associated risk factors on post-operative outcomes in children with CHDs underwent cardiac surgery. Methods We conducted a retrospective cohort study in a single center tertiary pediatric cardiac intensive care unit (PCICU) in Indonesia. Our cohort included all children between 5 and 36 months of age undergoing congenital heart surgery with cardiopulmonary bypass from November 2011 until February 2014. Outcomes measured were the length of intubation and the length of ICU stay. Variables for potential influence investigated were the nutritional status, age, gender, type of cardiac anomaly (acyanotic vs. cyanotic), Aristotle score, cardiopulmonary bypass time, aortic cross-clamp time, and Pediatric Risk of Mortality (PRISM) III score. Results Out of 249 patients included, 147 (59%) showed malnourishment on admission. Malnourished patients were significantly younger in age, presented with an acyanotic heart defects, and had higher PRISM III score. Additionally, they also had a longer mechanical ventilation time and ICU stay than those with a normal nutritional status. After adjusting for various variables using a multiple logistic regression model it could be demonstrated that a higher Z-score for weight to age was a significant protective factor for the intubation time of more than 29 hours with an odds ratio of 0.66 (95% CI 0.48 to 0.92, P = 0.012). Non-malnourished patients had a 49% significantly higher chance for extubation with a hazard ratio of 1.49 (95% CI 1.12 to 1.99, P= 0.007). Conclusion Malnourishment is clearly associated in a linear fashion with longer mechanical ventilation and ICU stay. As one of significant and potentially treatable co-morbidity factors, prevention of malnourishment by early diagnosis and optimal timing for surgery is important

Development and Utility of Quality Metrics for Ambulatory Pediatric Cardiology in Kawasaki Disease.

The Adult Congenital and Pediatric Cardiology (ACPC) Section of the American College of Cardiology sought to develop quality indicators/metrics for ambulatory pediatric cardiology practice. The objective of this study was to report the creation of metrics for patients with Kawasaki disease. Over a period of 5 months, 12 pediatric cardiologists developed 24 quality metrics based on the most relevant statements, guidelines, and research studies pertaining to Kawasaki disease. Of the 24 metrics, the 8 metrics deemed the most important, feasible, and valid were sent on to the ACPC for consideration. Seven of the 8 metrics were approved using the RAND method by an expert panel. All 7 metrics approved by the ACPC council were accepted by ACPC membership after an "open comments" process. They have been disseminated to the pediatric cardiology community for implementation by the ACPC Quality Network

MuRF1 mono-ubiquitinates TRα to inhibit T3-induced cardiac hypertrophy in vivo

Thyroid hormone (TH) is recognized for its role in cellular metabolism and growth and participates in homeostasis of the heart. T3 activates pro-survival pathways including Akt and mTOR. Treatment with T3 after myocardial infarction is cardioprotective and promotes elements of physiological hypertrophic response after cardiac injury. Although T3 is known to benefit the heart, very little about its regulation at the molecular level has been described to date. The ubiquitin proteasome system (UPS) regulates nuclear hormone receptors such as estrogen, progesterone, androgen, and glucocorticoid receptors by both degradatory and non-degradatory mechanisms. However, how the UPS regulates T3-mediated activity is not well understood. In this study, we aim to determine the role of the muscle-specific ubiquitin ligase muscle ring finger-1 (MuRF1) in regulating T3-induced cardiomyocyte growth. An increase in MuRF1 expression inhibits T3-induced physiological cardiac hypertrophy, whereas a decrease in MuRF1 expression enhances T3’s activity both in vitro and in cardiomyocytes in vivo. MuRF1 interacts directly with TRα to inhibit its activity by posttranslational ubiquitination in a non-canonical manner. We then demonstrated that a nuclear localization apparatus that regulates/inhibits nuclear receptors by sequestering them within a subcompartment of the nucleus was necessary for MuRF1 to inhibit T3 activity. This work implicates a novel mechanism that enhances the beneficial T3 activity specifically within the heart, thereby offering a potential target to enhance cardiac T3 activity in an organ-specific manner

Embedding a Native State into a Random Heteropolymer Model: The Dynamic Approach

We study a random heteropolymer model with Langevin dynamics, in the supersymmetric formulation. Employing a procedure similar to one that has been used in static calculations, we construct an ensemble in which the affinity of the system for a native state is controlled by a "selection temperature" T0. In the limit of high T0, the model reduces to a random heteropolymer, while for T0-->0 the system is forced into the native state. Within the Gaussian variational approach that we employed previously for the random heteropolymer, we explore the phases of the system for large and small T0. For large T0, the system exhibits a (dynamical) spin glass phase, like that found for the random heteropolymer, below a temperature Tg. For small T0, we find an ordered phase, characterized by a nonzero overlap with the native state, below a temperature Tn \propto 1/T0 > Tg. However, the random-globule phase remains locally stable below Tn, down to the dynamical glass transition at Tg. Thus, in this model, folding is rapid for temperatures between Tg and Tn, but below Tg the system can get trapped in conformations uncorrelated with the native state. At a lower temperature, the ordered phase can also undergo a dynamical glass transition, splitting into substates separated by large barriers.Comment: 19 pages, revtex, 6 figure