On a wing and a prayer: an assessment of modularized crew resource management training for health care professionals.
Evidence suggests that Crew Resource Management (CRM), a form of team training, is beneficial. In CRM training, participants learn individual portable team skills such as communication and decision making through group discussion and activities. However, the usual 1-day course format is not always compatible with health care organizational routines. A modular training format, while theoretically sound, is untested for interprofessional team training. The aim of this study was to explore the potential for modularized CRM training to be delivered to a group of interprofessional learners. Modularized CRM training, consisting of two 2-hour workshops, was delivered to health care workers in an Australian tertiary hospital. Kirkpatrick's evaluation model provided a framework for the study. Baseline attitude surveys were conducted prior to each workshop. Participants completed a written questionnaire at the end of each workshop that examined their motivations, reactions to the training, and learner demographics. An additional survey, administered 6 weeks post training, captured self-assessed behavior data. Twenty-three individuals from a range of professions and clinical streams participated. One in 5 participants (22%) reported that they translated teamwork skills to the workplace. While positive about the workshop format and content, many respondents identified personal, team, and organizational barriers to the application of the workshop techniques. CRM training when delivered in a modular format has positive outcomes. Following the training, some respondents overcame workplace barriers to attempt to change negative workplace behavior. This progress provides cautious optimism for the potential for modular CRM training to benefit groups of interprofessional health staff.


Early prospective clinical results of a modern fixed-bearing total ankle arthroplasty.
Several fixed-bearing total ankle arthroplasty systems are available in the United States. We report on the early clinical results of the largest known cohort of patients in the United States who received a Salto Talaris total ankle replacement for the treatment of end-stage arthritis of the ankle. We prospectively followed sixty-seven patients with a minimum clinical follow-up of two years. Patients completed standardized assessments and underwent physical examination, functional assessment, and radiographic evaluation preoperatively and at six weeks, three months, and six months postoperatively and yearly thereafter through their most recent follow-up. Implant survival at a mean follow-up time of 2.8 years was 96% when metallic component revision, removal, or impending failure was used as the end point. Three patients developed aseptic loosening, and all instances involved the tibial component. One of the three patients underwent revision to another fixed-bearing total ankle arthroplasty system, one patient is awaiting revision surgery, and the third patient has remained minimally symptomatic and fully functional without additional surgery. Forty-five patients underwent at least one additional procedure at the time of the index surgery. The most common concurrent procedure performed was a deltoid ligament release (n = 21). Eight patients underwent additional surgery following the index arthroplasty, most commonly debridement for medial and/or lateral impingement (n = 4). As of the most recent follow-up, patients demonstrated significant improvement in pain scores, American Orthopaedic Foot & Ankle Society hindfoot score, and functional scores. Early clinical results indicate that the Salto Talaris fixed-bearing total ankle arthroplasty system can provide significant improvement in pain, quality of life, and standard functional measures in patients with end-stage ankle arthritis. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Cryptic nature of envelope V3 region epitopes protects primary monocytotropic human immunodeficiency virus type 1 from antibody neutralization.
Characterization of biological and immunological properties of human immunodeficiency virus type 1 (HIV-1) is critical to developing effective therapies and vaccines for AIDS. With the use of a novel CD4+ T-cell line (PM-1) permissive to infection by both monocytotropic (MT) and T-cell-tropic virus types, we present a comparative analysis of the immunological properties of a prototypic primary MT isolate of HIV-1 strain JR-CSF (MT-CSF) with those of a T-cell-tropic variant (T-CSF) of the same virus, which emerged spontaneously in vitro. The parental MT-CSF infected only PM-1 cells and was markedly resistant to neutralization by sera from HIV-1-infected individuals, rabbit antiserum to recombinant MT-CSF gp120, and anti-V3 monoclonal antibodies. The T-CSF variant infected a variety of CD4+ T-cell lines, contained positively charged amino acid substitutions in the gp120 V3 region, and was highly sensitive to antibody neutralization. Neutralization and antibody staining of T-CSF-expressing cells were significantly inhibited by HIV-1 V3 peptides; in contrast, the MT strain showed only weak V3-specific binding of polyclonal and monoclonal antibodies. Exposure of PM-1 cells to a mixture of both viruses in the presence of human anti-HIV-1 neutralizing antiserum resulted in infection with only MT-CSF. These results demonstrate that although the V3 region of MT viruses is immunogenic, the target epitopes in the V3 principal neutralizing domain on the membrane form of the MT envelope appear to be cryptic or hidden from blocking antibodies.


The Hippo pathway regulates the bantam microRNA to control cell proliferation and apoptosis in Drosophila.
The Hippo signaling pathway acts upon the Yorkie transcriptional activator to control tissue growth in Drosophila. Activated Yorkie drives growth by stimulating cell proliferation and inhibiting apoptosis, but how it achieves this is not understood. Yorkie is known to activate Cyclin E (CycE) and the apoptosis inhibitor DIAP1. However, overexpression of these targets is not sufficient to cause tissue overgrowth. Here we show that Yorkie also activates expression of the bantam microRNA, a known regulator of both proliferation and apoptosis. bantam overexpression mimics Yorkie activation while loss of bantam function slows the rate of cell proliferation. bantam is necessary for Yorkie-induced overproliferation and bantam overexpression is sufficient to rescue survival and proliferation of yorkie mutant cells. Finally, we show that bantam levels are regulated during both developmentally programmed proliferation arrest and apoptosis. In summary, the results show that the Hippo pathway regulates expression of bantam to control tissue growth in Drosophila.


Why Girls Choose Not to Use Barriers to Prevent Sexually Transmitted Infection During Female-to-Female Sex.
Using data from a national qualitative study of lesbian, bisexual, and other sexual minority adolescent girls in the U.S., this study examined their awareness of the risk of sexually transmitted infection (STI) and opportunities for barrier use. Online asynchronous focus groups were conducted with lesbian and bisexual (LB) girls ages 14-18 years. Girls were assigned to online groups based on their self-identified sexual identity and whether they were sexually experienced or not. Two moderators posed questions and facilitated online discussions. Interpretive description analysis conducted by multiple members of the research team was used to categorize the results. Key factors in girls' decisions not to use barriers with female partners concerned pleasure, sex of sexual partner, lack of knowledge of sexual risk or of barrier use for female-to-female sexual activities, and use of STI testing as a prevention tool. Addressing knowledge and access gaps is an important first step for improving sexual health. Prevention priorities should focus on helping LB girls understand their risk of STI transmission in both opposite and same-sex relationships. Tailoring messaging to move beyond heteronormative scripts is critical to engaging LB girls and equipping them with the skills and knowledge to have safer sex regardless of the sex of their partner.


Efficacy of Perioperative Lumbar Drainage following Endonasal Endoscopic Cerebrospinal Fluid Leak Repair.
Objective Perioperative lumbar drain (LD) use in the setting of endoscopic cerebrospinal fluid (CSF) leak repair is a well-established practice. However, recent data suggest that LDs may not provide significant benefit and may thus confer unnecessary risk. To examine this, we conducted a meta-analysis to investigate the effect of LDs on postoperative CSF leak recurrence following endoscopic repair of CSF rhinorrhea. Data Sources A comprehensive search was performed with the following databases: Ovid MEDLINE (1947 to November 2015), EMBASE (1974 to November 2015), Cochrane Review, and PubMed (1990 to November 2015). Review Method A meta-analysis was performed according to PRISMA guidelines. Results A total of 1314 nonduplicate studies were identified in our search. Twelve articles comprising 508 cases met inclusion criteria. Overall, use of LDs was not associated with significantly lower postoperative CSF leak recurrence rates following endoscopic repair of CSF rhinorrhea (odds ratio: 0.89, 95% confidence interval: 0.40-1.95) as compared with cases performed without LDs. Subgroup analysis of only CSF leaks associated with anterior skull base resections (6 studies, 153 cases) also demonstrated that lumbar drainage did not significantly affect rates of successful repair (odds ratio: 2.67, 95% confidence interval: 0.64-11.10). Conclusions There is insufficient evidence to support that adjunctive lumbar drainage significantly reduces postoperative CSF leak recurrence in patients undergoing endoscopic CSF leak repair. Subgroup analysis examining only those patients whose CSF leaks were associated with anterior skull base resections demonstrated similar results. More level 1 and 2 studies are needed to further investigate the efficacy of LDs, particularly in the setting of patients at high risk for CSF leak recurrence.


Heads or tails: do stranded fish (mosquitofish, Gambusia affinis) know where they are on a slope and how to return to the water?
Aquatic vertebrates that emerge onto land to spawn, feed, or evade aquatic predators must return to the water to avoid dehydration or asphyxiation. How do such aquatic organisms determine their location on land? Do particular behaviors facilitate a safe return to the aquatic realm? In this study, we asked: will fully-aquatic mosquitofish (Gambusia affinis) stranded on a slope modulate locomotor behavior according to body position to facilitate movement back into the water? To address this question, mosquitofish (n = 53) were placed in four positions relative to an artificial slope (30° inclination) and their responses to stranding were recorded, categorized, and quantified. We found that mosquitofish may remain immobile for up to three minutes after being stranded and then initiate either a "roll" or a "leap". During a roll, mass is destabilized to trigger a downslope tumble; during a leap, the fish jumps up, above the substrate. When mosquitofish are oriented with the long axis of the body at 90° to the slope, they almost always (97%) initiate a roll. A roll is an energetically inexpensive way to move back into the water from a cross-slope body orientation because potential energy is converted back into kinetic energy. When placed with their heads toward the apex of the slope, most mosquitofish (>50%) produce a tail-flip jump to leap into ballistic flight. Because a tail-flip generates a caudually-oriented flight trajectory, this locomotor movement will effectively propel a fish downhill when the head is oriented up-slope. However, because the mass of the body is elevated against gravity, leaps require more mechanical work than rolls. We suggest that mosquitofish use the otolith-vestibular system to sense body position and generate a behavior that is "matched" to their orientation on a slope, thereby increasing the probability of a safe return to the water, relative to the energy expended.


Visual Category Learning Results in Rapid Changes in Brain Activation Reflecting Sensitivity to the Category Relation between Perceived Objects and to Decision Correctness.
Little is known about the time scales in which sensitivity to novel category identity may become evident in visual and executive cortices in visual category learning (VCL) tasks and the nature of such changes in brain activation. We used fMRI to investigate the processing of category information and trial-by-trial feedback information. In each VCL task, stimuli differed in three feature dimensions. In each trial, either two same-category stimuli or two different-categories stimuli were presented. The participant had to learn which feature dimension was relevant for categorization based on the feedback that followed each categorization decision. We contrasted between same-category stimuli trials and different-category trials and between correct and incorrect categorization decision trials. In each trial, brain activation in the visual stimuli processing phase was modeled separately from activation during the later feedback processing phase. We found activation in the lateral occipital complex, indicating sensitivity to the category relation between stimuli, to be evident in VCL within only few learning trials. Specifically, greater lateral occipital complex activation was evident when same-category stimuli were presented than when different-category stimuli were presented. In the feedback processing phase, greater activation in both executive and visual cortices was evident primarily after "misdetections" of same-category stimuli. Implications regarding the contribution of different learning trials to VCL, and the respective role of key brain regions, at the onset of VCL, are discussed.


Serious Games and Gamification for Mental Health: Current Status and Promising Directions.
Computer games are ubiquitous and can be utilized for serious purposes such as health and education. "Applied games" including serious games (in brief, computerized games for serious purposes) and gamification (gaming elements used outside of games) have the potential to increase the impact of mental health internet interventions via three processes. First, by extending the reach of online programs to those who might not otherwise use them. Second, by improving engagement through both game-based and "serious" motivational dynamics. Third, by utilizing varied mechanisms for change, including therapeutic processes and gaming features. In this scoping review, we aim to advance the field by exploring the potential and opportunities available in this area. We review engagement factors which may be exploited and demonstrate that there is promising evidence of effectiveness for serious games for depression from contemporary systematic reviews. We illustrate six major categories of tested applied games for mental health (exergames, virtual reality, cognitive behavior therapy-based games, entertainment games, biofeedback, and cognitive training games) and demonstrate that it is feasible to translate traditional evidence-based interventions into computer gaming formats and to exploit features of computer games for therapeutic change. Applied games have considerable potential for increasing the impact of online interventions for mental health. However, there are few independent trials, and direct comparisons of game-based and non-game-based interventions are lacking. Further research, faster iterations, rapid testing, non-traditional collaborations, and user-centered approaches are needed to respond to diverse user needs and preferences in rapidly changing environments.


The prokinetic face of ghrelin.
This review evaluated published data regarding the effects of ghrelin on GI motility using the PubMed database for English articles from 1999 to September 2009. Our strategy was to combine all available information from previous literature, in order to provide a complete structured review on the prokinetic properties of exogenous ghrelin and its potential use for treatment of various GI dysmotility ailments. We classified the literature into two major groups, depending on whether studies were done in health or in disease. We sub-classified the studies into stomach, small intestinal and colon studies, and broke them down further into studies done in vitro, in vivo (animals) and in humans. Further more, the reviewed studies were presented in a chronological order to guide the readers across the scientific advances in the field. The review shows evidences that ghrelin and its (receptor) agonists possess a strong prokinetic potential to serve in the treatment of diabetic, neurogenic or idiopathic gastroparesis and possibly, chemotherapy-associated dyspepsia, postoperative, septic or post-burn ileus, opiate-induced bowel dysfunction and chronic idiopathic constipation. Further research is necessary to close the gap in knowledge about the effect of ghrelin on the human intestines in health and disease.


Risk factors for the presence of dengue vector mosquitoes, and determinants of their prevalence and larval site selection in Dhaka, Bangladesh.
Dengue viruses are responsible for over 100 million infections a year worldwide and are a public health concern in Bangladesh. Although risk of transmission is high, data on vector population characteristics are scanty in Bangladesh; therefore, a comprehensive prediction of the patterns of local virus transmission is not possible. Recognizing these gaps, multi-year entomological surveys were carried out in Dhaka, where the disease is most frequently reported. The specific objectives of the present study are threefold: i) to determine the risk factors for the presence of Aedes mosquitoes; ii) to identify the types of most productive and key containers; and iii) to estimate the effects of climatic factors on Aedes abundance in the city of Dhaka, Bangladesh. Entomological surveys were conducted in 12 out of 90 wards in Dhaka. These wards were selected using a probability proportional sampling procedure during the monsoon seasons in 2011, 2012 and 2013 and in the dry season in 2012. All containers inside and around sampled households were inspected for mosquito larvae and pupae, and containers were classified according to their relative size, use pattern, and materials of construction. During the study period (2011-2013), 12,680 larvae and pupae were collected. About 82% of the identified immature mosquitoes were Aedes aegypti, while the remainder were Ae. albopictus and other mosquito species. The largest number of immature mosquitoes was collected from tires and refrigerator trays during 2011 and 2012 monsoon seasons. Conversely, plastic drums were the most productive during the 2012 dry and 2013 monsoon season. Vehicle parts and discarded construction materials were the most efficient producers of Aedes mosquitoes in all surveys. The presence of Aedes mosquitoes was significantly (p < 0.05) higher in low socio-economic zones of Dhaka. Container location, presence of vegetation, and availability of shade for containers were also significantly associated with finding immature Aedes mosquitoes, based on multivariable analysis after confounder adjustment. Rainfall, temperature, and relative humidity also significantly affected the mean abundance of mosquitoes. Proper use, disposal, and recycling of the containers that effectively produce large numbers of Aedes vector mosquitoes may decrease the risk of arboviral transmission.


Sonographic evaluation of the breast.
Knowledge of appropriate ultrasonographic (US) techniques and the US appearance of the normal breast and specific lesions is essential for successful application of breast US. The highest quality breast sonograms are achieved with 5-10-MHz linear transducers, imaging the lesion within the transducer focal zone, and relatively steep time-gain compensation curves. On sonograms, the fat in normal breast parenchyma is hypoechoic, fibrous tissue is echogenic, and glandular tissue is intermediate in echogenicity. Some normal structures can simulate masses, including the anterior costochondral junction when imaged in cross section and fat lobules outlined by Cooper ligaments when imaged in a perpendicular plane. Cysts have an anechoic interior, sharp margins, and posterior acoustic enhancement. Benign solid lesions are usually hypoechoic but variable in US appearance, which can overlap with that of complicated cysts. The classic US appearance of breast carcinoma is a hypoechoic mass with inhomogeneous internal echoes, irregular margins, and variable acoustic shadowing, although carcinoma can appear well circumscribed or have posterior acoustic shadowing in some cases. The most important function of breast US is differentiating a cyst from a solid lesion. US is also useful for evaluating a palpable mass in either young patients (< 30 years old) or those with dense breasts and negative mammographic results.


Accuracies of the diffusion approximation and its similarity relations for laser irradiated biological media.
The accuracy of the diffusion approximation is compared with more accurate solutions for describing light interaction with biological tissues. Generally the diffusion approximation underestimates the light distribution in the surface region, and, for high albedos, it significantly underestimates the fluence rate. This difference is only a few percent for albedos of less than 0.5 due to the dominance of collimated light. As the anisotropy of scattering increases, deviations increase. In general, fluxes can be computed more accurately with the diffusion approximation than fluence rates. For anisotropic scattering, better results can be obtained by simple transforms of optical coefficients using the similarity relations. The similarity relations improve flux calculations, but computed fluence rates have substantial errors for high albedo and the large index of refraction differences at the surface.


The role of individual susceptibility in cancer burden related to environmental exposure.
Individual susceptibility to cancer may result from host factors including differences n metabolism, DNA repair, altered expression of protooncogenes and tumor suppressor genes, and nutritional status. Since most carcinogens require metabolic activation before binding to DNA, variations in an individual's metabolic phenotype that have detected in enzymes involved in activation and detoxification should play an essential role in the development of environmental cancer. This phenotypic metabolic variation has now been related to genetic polymorphisms, and many genes encoding carcinogen-metabolizing enzymes have been identified and cloned. Consequently, allelic variants or genetic defects that give rise to the observed variation and new polymorphisms have been recognized. Development of simple polymerase chain reaction (PCR)-based assays has enabled identification of an individual's genotype for a variety of metabolic polymorphisms. Thus, recent knowledge of the genetic basis for individual metabolic variation has opened new possibilities of studies focusing on increased individual susceptibility to environmentally induced cancer, which are reviewed with special reference to smoking-induced lung cancer. Cancer susceptibility due to chemical exposure is likely to be determined by an individual's phenotype for a number of enzymes (both activating and detoxifying) relevant to that of a single carcinogen or mixtures of carcinogens. Given the number and variability in expression of carcinogen-metabolizing enzymes and the complexity of chemical exposures, assessment of a single polymorphic enzyme (genotype) may not be sufficient. Mutations in the p53 gene are among the most common genetic changes in human cancer. The frequency and type p53 mutations can act as a fingerprint of carcinogen exposure and may therefore provide information about external etiological agents, intensity of exposure, and host factors affecting the tumorigenesis process. In human lung cancer, p53 mutations (both the mutation pattern and frequency) have been linked with tobacco smoking; the type of mutation most frequently observed is G:C to T:A transversion, a mutation preferentially induced by benzo[a]pyrene diol epoxide. An association between the presence of this transversion and the genotype deficient in glutathione S-transferase M1-mediated detoxification has been observed in lung cancer. Taken together, these findings suggest that determination of metabolic at risk genotypes in combination with levels of DNA adducts in target (surrogate) tissues and the p53 mutation pattern should allow the identification of susceptible individuals and subgroups in carcinogen-exposed populations.


Small intestinal metabolism is central to whole-body insulin resistance.
To assess the role of jejunum in insulin resistance in humans and in experimental animals. Twenty-four subjects undergoing biliopancreatic diversion (BPD) or Roux-en-Y gastric bypass (RYGB) were enrolled. Insulin sensitivity was measured at baseline and at 1 week after surgery using oral glucose minimal model.We excluded the jejunum from intestinal continuity in pigs and created a jejunal loop with its vascular and nerve supply intact accessible from two cutaneous stomas, and reconnected the bowel with an end-to-end anastomosis. Glucose stable isotopes were given in the stomach or in the jejunal loop.In vitro studies using primary porcine and human hepatocytes or myoblasts tested the effects of plasma on gluconeogenesis or glucose uptake and insulin signalling. Whole-body insulin sensitivity (SI∙104: 0.54±0.12 before vs 0.82±0.11 after BPD, p=0.024 and 0.41±0.09 before vs 0.65±0.09/pM/min after RYGB, p=not significant) and Glucose Disposition Index increased only after BPD. In pigs, insulin sensitivity was significantly lower when glucose was administered in the jejunal loop than in the stomach (glucose rate of disappearance (Rd) area under the curve (AUC)/insulin AUC∙10: 1.82±0.31 vs 2.96±0.33 mmol/pM/min, p=0.0017).Metabolomics showed a similar pattern before surgery and during jejunal-loop stimulation, pointing to a higher expression of gluconeogenetic substrates, a metabolic signature of impaired insulin sensitivity.A greater hepatocyte phosphoenolpyruvate-carboxykinase and glucose-6-phosphatase gene expression was elicited with plasma from porcine jejunal loop or before surgery compared with plasma from jejunectomy in pigs or jejunal bypass in humans.Stimulation of myoblasts with plasma from porcine jejunal loop or before surgery reduced glucose uptake, Ser473-Akt phosphorylation and GLUT4 expression compared with plasma obtained during gastric glucose administration after jejunectomy in pigs or after jejunal bypass in humans. Proximal gut plays a crucial role in controlling insulin sensitivity through a distinctive metabolic signature involving hepatic gluconeogenesis and muscle insulin resistance. Bypassing the jejunum is beneficial in terms of insulin-mediated glucose disposal in obesity. NCT03111953.


Primary aldosteronism: renaissance of a syndrome.
Great strides have been made in our understanding of the pathophysiology of primary aldosteronism syndrome since Conn's description of the clinical presentation of a patient with an aldosterone-producing adenoma (APA) more than 50 years ago. It is now recognized that the APA is just one of the seven subtypes of primary aldosteronism. APA and bilateral idiopathic hyperaldosteronism (IHA) are the most common subtypes of primary aldosteronism. Although most clinicians had thought primary aldosteronism to be a rare form of hypertension for more than three decades, it is now recognized to be the most common form of secondary hypertension. Using the plasma aldosterone to plasma renin activity ratio as a case-finding test, followed by aldosterone suppression confirmatory testing, has resulted in much higher prevalence estimates of 5-13% of all patients with hypertension. In addition, there has been a new recognition of the aldosterone-specific cardiovascular morbidity and mortality associated with aldosterone excess. Although thought to be daunting and complex in the past, the diagnostic approach to primary aldosteronism is straightforward and can be considered in three phases: case-finding tests, confirmatory tests and subtype evaluation tests. Patients with hypertension and hypokalaemia (regardless of presumed cause), treatment-resistant hypertension (three antihypertensive drugs and poor control), severe hypertension (>or= 160 mmHg systolic or >or= 100 mmHg diastolic), hypertension and an incidental adrenal mass, onset of hypertension at a young age or patients being evaluated for other forms of secondary hypertension should undergo screening for primary aldosteronism. In patients with suspected primary aldosteronism, screening can be accomplished by measuring a morning (preferably between 0800 and 1000 h) ambulatory paired random plasma aldosterone concentration (PAC) and plasma renin activity (PRA). An increased PAC:PRA ratio is not diagnostic by itself, and primary aldosteronism must be confirmed by demonstrating inappropriate aldosterone secretion. Aldosterone suppression testing can be performed with orally administered sodium chloride and measurement of urinary aldosterone or with intravenous sodium chloride loading and measurement of PAC. Unilateral adrenalectomy in patients with APA or unilateral adrenal hyperplasia results in normalization of hypokalaemia in all these patients; hypertension is improved in all and is cured in approximately 30-60% of them. In bilateral adrenal forms of primary aldosteronism, unilateral or bilateral adrenalectomy seldom corrects the hypertension and they should be treated medically with a mineralocorticoid receptor antagonist.


Individual right whales call louder in increased environmental noise.
The ability to modify vocalizations to compensate for environmental noise is critical for successful communication in a dynamic acoustic environment. Many marine species rely on sound for vital life functions including communication, navigation and feeding. The impacts of significant increases in ocean noise levels from human activities are a current area of concern for the conservation of marine mammals. Here, we document changes in calling behaviour by individual endangered North Atlantic right whales (Eubalaena glacialis) in increased background noise. Right whales, like several bird and primate species, respond to periods of increased noise by increasing the amplitude of their calls. This behaviour may help maintain the communication range with conspecifics during periods of increased noise. These call modifications have implications for conservation efforts for right whales, affecting both the way whales use sound to communicate and our ability to detect them with passive acoustic monitoring systems.


The cubic-to-hexagonal phase transition of cerium oxide particles: dynamics and structure.
Cerium oxide is often applied in today's catalysts due to its remarkable oxygen storage capacity. The changes in stoichiometry during reaction are linked to structural modifications, which in turn affect its catalytic activity. We present a real-time in situ study of the structural transformations of cerium oxide particles on ruthenium(0001) at high temperatures of 700 °C in ultra-high vacuum. Our results demonstrate that the reduction from CeO2 to cubic Ce2O3 proceeds via ordered intermediary phases. The final reduction step from cubic to hexagonal Ce2O3 is accompanied by a lattice expansion, the formation of two new surface terminations, a partial dissolution of the cerium oxide particles, and a massive mass transport of cerium from the particles to the substrate. The conclusions allow for new insights into the structure, stability, and dynamics of cerium oxide nanoparticles in strongly reducing environments.


Improved secondary structure predictions for a nicotinic receptor subunit: incorporation of solvent accessibility and experimental data into a two-dimensional representation.
Abstract A refined prediction of the nicotinic acetylcholine receptor (nAChR) subunits' secondary structure was computed with third-generation algorithms. The four selected programs, PHD, Predator, DSC, and NNSSP, based on different prediction approaches, were applied to each sequence of an alignment of nAChR and 5-HT3 receptor subunits, as well as a larger alignment with related subunit sequences from glycine and GABA receptors. A consensus prediction was computed for the nAChR subunits through a "winner takes all" method. By integrating the probabilities obtained with PHD, DSC, and NNSSP, this prediction was filtered in order to eliminate the singletons and to more precisely establish the structure limits (only 4% of the residues were modified). The final consensus secondary structure includes nine alpha-helices (24.2% of the residues, with an average length of 13.9 residues) and 17 beta-strands (22.5% of the residues, with an average length of 6.6 residues). The large extracellular domain is predicted to be mainly composed of beta-strands, with only two helices at the amino-terminal end. The transmembrane segments are predicted to be in a mixed alpha/beta topology (with a predominance of alpha-helices), with no known equivalent in the current protein database. The cytoplasmic domain is predicted to consist of two well-conserved amphipathic helices joined together by an unfolded stretch of variable length and sequence. In general, the segments predicted to occur in a periodic structure correspond to the more conserved regions, as defined by an analysis of sequence conservation per position performed on 152 superfamily members. The solvent accessibility of each residue was predicted from the multiple alignments with PHDacc. Each segment with more than three exposed residues was assumed to be external to the core protein. Overall, these data constitute an envelope of structural constraints. In a subsequent step, experimental data relative to the extracellular portion of the complete receptor were incorporated into the model. This led to a proposed two-dimensional representation of the secondary structure in which the peptide chain of the extracellular domain winds alternatively between the two interfaces of the subunit. Although this representation is not a tertiary structure and does not lead to predictions of specific beta-beta interaction, it should provide a basic framework for further mutagenesis investigations and for fold recognition (threading) searches.


Toxoplasma from the eggs of the domestic fowl (Gallus gallus).
Toxoplasmata of varying grades of virulence were isolated from eggs laid by clinically asymptomatic hens and from their internal organs. The strains so recovered were identified as Toxoplasna gondii by morphological, serological, cultural, and pathogenic criteria. These findings strongly suggest that toxoplasmosis may be contracted through the eating of raw or undercooked infected eggs.


Quantifying self-absorption losses in luminescent solar concentrators.
Analytical equations quantifying self-absorption losses in circular luminescent solar concentrators (LSCs) are presented that can easily be solved numerically by commercial math software packages. With the quantum efficiency, the absorption and emission spectra of a luminescent material, the LSC dimensions, and the refractive index as the only input parameters, the model gives an accurate account of the decrease of LSC efficiency due to self-absorption as a function of LSC radius, thickness, and luminescence quantum efficiency. Results give insight into how many times light is reabsorbed and reemitted, the red shift of the emission spectrum, and on how multiple reabsorptions and reemissions are distributed over the LSC. As an example case the equations were solved for a circular LSC containing a Lumogen F Red 305 dye with 80% luminescence quantum efficiency, and it follows that for an LSC with a 50 cm radius the self-absorption reduces the number of photons reaching the LSC edge by a factor of four compared to the case when there would be no self-absorption. The equations can just as well be solved for any material for which the optical properties are known like type I and type II quantum dots.


Impact of Refugees on Local Health Systems: A Difference-in-Differences Analysis in Cameroon.
Hosting refugees may represent a drain on local resources, particularly since external aid is frequently insufficient. Between 2004 and 2011, over 100,000 refugees settled in the eastern border of Cameroon. With little known on how refugee influx affects health services of the hosting community, we investigated the impact of refugees on mother and child health (MCH) services in the host community in Cameroon. We used Cameroon's 2004 and 2011 Demographic and Health Surveys to evaluate changes in MCH indicators in the refugee hosting community. Our outcome variables were antenatal care (ANC) coverage, caesarean delivery rate, place of delivery and child vaccination coverage; whereas the exposure variable was residence in the refugee hosting community. We used a difference-in-differences analysis to compare indicators of the refugee hosting community to a control group selected through propensity score matching from the rest of the country. A total of 10,656 women were included in our 2004 analysis and 7.6% (n = 826) of them resided in the refugee hosting community. For 2011, 15,426 women were included and 5.8% (n = 902) of them resided in the hosting community. Between 2004 and 2011, both the proportion of women delivering outside health facilities and children not completing DPT3 vaccination in the refugee hosting community decreased by 9.0% (95% Confidence Interval (CI): 3.9-14.1%) and 9.6% (95% CI: 7.9-11.3%) respectively. However, ANC attendance and caesarean delivery did not show any significant change. Our findings demonstrate that none of the evaluated MCH service indicators deteriorated (in fact, two of them improved: delivery in health facilities and completing DPT3 vaccine) with the presence of refugees. This suggests evidence disproving the common belief that refugees always have a negative impact on their hosting community.


BP-Dock: a flexible docking scheme for exploring protein-ligand interactions based on unbound structures.
Molecular docking serves as an important tool in modeling protein-ligand interactions. However, it is still challenging to incorporate overall receptor flexibility, especially backbone flexibility, in docking due to the large conformational space that needs to be sampled. To overcome this problem, we developed a novel flexible docking approach, BP-Dock (Backbone Perturbation-Dock) that can integrate both backbone and side chain conformational changes induced by ligand binding through a multi-scale approach. In the BP-Dock method, we mimic the nature of binding-induced events as a first-order approximation by perturbing the residues along the protein chain with a small Brownian kick one at a time. The response fluctuation profile of the chain upon these perturbations is computed using the perturbation response scanning method. These response fluctuation profiles are then used to generate binding-induced multiple receptor conformations for ensemble docking. To evaluate the performance of BP-Dock, we applied our approach on a large and diverse data set using unbound structures as receptors. We also compared the BP-Dock results with bound and unbound docking, where overall receptor flexibility was not taken into account. Our results highlight the importance of modeling backbone flexibility in docking for recapitulating the experimental binding affinities, especially when an unbound structure is used. With BP-Dock, we can generate a wide range of binding site conformations realized in nature even in the absence of a ligand that can help us to improve the accuracy of unbound docking. We expect that our fast and efficient flexible docking approach may further aid in our understanding of protein-ligand interactions as well as virtual screening of novel targets for rational drug design.


Chronic Myelomonocytic Leukemia: Insights into Biology, Prognostic Factors, and Treatment.
Chronic myelomonocytic leukemia (CMML) is a clonal hematological malignancy characterized by both dysplastic and proliferative features, with an inherent risk for leukemic transformation. With the help of this review, we aim to summarize key concepts with regards to CMML biology, diagnosis, risk stratification, and therapeutics. Based on recent studies, CMML is hallmarked by a relatively low genetic complexity, which contrasts with a compelling phenotypical heterogeneity, largely driven by epigenetic mechanisms. Recent advances in the characterization of CMML biology has led to an improvement in risk-stratification, by means of incorporating prognostically relevant gene mutations. This, however, has not significantly impacted available therapies and outcomes continue to remain poor. Advances in CMML biology have better explained the phenotypic heterogeneity, while continuing to define the genetic and epigenetic landscape. In spite of recent advances, limited effective therapies exist and developing rationally derived therapeutic approaches is much needed.


Literature-aided interpretation of gene expression data with the weighted global test.
Most methods for the interpretation of gene expression profiling experiments rely on the categorization of genes, as provided by the Gene Ontology (GO) and pathway databases. Due to the manual curation process, such databases are never up-to-date and tend to be limited in focus and coverage. Automated literature mining tools provide an attractive, alternative approach. We review how they can be employed for the interpretation of gene expression profiling experiments. We illustrate that their comprehensive scope aids the interpretation of data from domains poorly covered by GO or alternative databases, and allows for the linking of gene expression with diseases, drugs, tissues and other types of concepts. A framework for proper statistical evaluation of the associations between gene expression values and literature concepts was lacking and is now implemented in a weighted extension of global test. The weights are the literature association scores and reflect the importance of a gene for the concept of interest. In a direct comparison with classical GO-based gene sets, we show that use of literature-based associations results in the identification of much more specific GO categories. We demonstrate the possibilities for linking of gene expression data to patient survival in breast cancer and the action and metabolism of drugs. Coupling with online literature mining tools ensures transparency and allows further study of the identified associations. Literature mining tools are therefore powerful additions to the toolbox for the interpretation of high-throughput genomics data.


A frontal attention mechanism in the visual mismatch negativity.
Automatic detection of environmental change is a core component of attention. The mismatch negativity (MMN), an electrophysiological marker of this mechanism, has been studied prominently in the auditory domain, with cortical generators identified in temporal and frontal regions. Here, we combined electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) to assess whether the underlying frontal regions associated with auditory change detection also play a role in visual change detection. Twenty healthy young adults completed a visual MMN task in separate EEG and fMRI sessions. Region of interest analyses were conducted on left and right middle frontal (MFG) and inferior frontal (IFG) gyri, i.e., the frontal areas identified as potential auditory MMN generators. A significant increase in activation was observed in the left IFG and MFG in response to blocks containing deviant stimuli. These findings suggest that a frontal mechanism is involved in the detection of change in the visual MMN. Our results support the notion that frontal mechanisms underlie attention switching, as measured via MMN, across multiple modalities.


siDirect: highly effective, target-specific siRNA design software for mammalian RNA interference.
siDirect (http://design.RNAi.jp/) is a web-based online software system for computing highly effective small interfering RNA (siRNA) sequences with maximum target-specificity for mammalian RNA interference (RNAi). Highly effective siRNA sequences are selected using novel guidelines that were established through an extensive study of the relationship between siRNA sequences and RNAi activity. Our efficient software avoids off-target gene silencing to enumerate potential cross-hybridization candidates that the widely used BLAST search may overlook. The website accepts an arbitrary sequence as input and quickly returns siRNA candidates, providing a wide scope of applications in mammalian RNAi, including systematic functional genomics and therapeutic gene silencing.


The Effects of Cucumber Mosaic Virus and Its 2a and 2b Proteins on Interactions of Tomato Plants with the Aphid Vectors Myzus persicae and Macrosiphum euphorbiae.
Cucumber mosaic virus (CMV), a major tomato pathogen, is aphid-vectored in the non-persistent manner. We investigated if CMV-induced volatile organic compounds (VOCs) or other virus-induced cues alter aphid-tomato interactions. Y-tube olfactometry showed that VOCs emitted by plants infected with CMV (strain Fny) attracted generalist (Myzus persicae) and Solanaceae specialist (Macrosiphum euphorbiae) aphids. Myzus persicae preferred settling on infected plants (3 days post-inoculation: dpi) at 1h post-release, but at 9 and 21 dpi, aphids preferentially settled on mock-inoculated plants. Macrosiphum euphorbiae showed no strong preference for mock-inoculated versus infected plants at 3 dpi but settled preferentially on mock-inoculated plants at 9 and 21 dpi. In darkness aphids showed no settling or migration bias towards either mock-inoculated or infected plants. However, tomato VOC blends differed in light and darkness, suggesting aphids respond to a complex mix of olfactory, visual, and other cues influenced by infection. The LS-CMV strain induced no changes in aphid-plant interactions. Experiments using inter-strain recombinant and pseudorecombinant viruses showed that the Fny-CMV 2a and 2b proteins modified tomato interactions with Macrosiphum euphorbiae and Myzus persicae, respectively. The defence signal salicylic acid prevents excessive CMV-induced damage to tomato plants but is not involved in CMV-induced changes in aphid-plant interactions.


Receptor-mediated endocytosis of epidermal growth factor by hepatocytes in the perfused rat liver: ligand and receptor dynamics.
We have used biochemical and morphological techniques to demonstrate that hepatocytes in the perfused liver bind, internalize, and degrade substantial amounts of murine epidermal growth factor (EGF) via a receptor-mediated process. Before ligand exposure, about 300,000 high-affinity receptors were detectable per cell, displayed no latency, and co-distributed with conventional plasma membrane markers. Cytochemical localization using EGF coupled to horseradish peroxidase (EGF-HRP) revealed that the receptors were distributed along the entire sinusoidal and lateral surfaces of hepatocytes. When saturating concentrations of EGF were perfused through a liver at 35 degrees C, ligand clearance was biphasic with a rapid primary phase of 20,000 molecules/min per cell that dramatically changed at 15-20 min to a slower secondary phase of 2,500 molecules/min per cell. During the primary phase of uptake, approximately 250,000 molecules of EGF and 80% of the total functional receptors were internalized into endocytic vesicles which could be separated from enzyme markers for plasma membranes and lysosomes on sucrose gradients. The ligand pathway was visualized cytochemically 2-25 min after EGF-HRP internalization and a rapid transport from endosomes at the periphery to those in the Golgi apparatus-lysosome region was observed (t 1/2 approximately equal to 7 min). However, no 125I-EGF degradation was detected for at least 20 min. Within 30 min after EGF addition, a steady state was reached which lasted up to 4 h such that (a) the rate of EGF clearance equaled the rate of ligand degradation (2,500 molecules/min per cell); (b) a constant pool of undegraded ligand was maintained in endosomes; and (c) the number of accessible (i.e., cell surface) receptors remained constant at 20% of initial values. By 4 h hepatocytes had internalized and degraded 3 and 2.3 times more EGF, respectively, than the initial number of available receptors, even in the presence of cycloheximide and without substantial loss of receptors. All of these results suggest that EGF receptors are internalized and that their rate of recycling to the surface from intracellular sites is governed by the rate of entry of ligand and/or receptor into lysosomes.


Reversible inactivation of dihydrolipoamide dehydrogenase by mitochondrial hydrogen peroxide.
Under oxidative stress conditions, mitochondria are the major site for cellular production of reactive oxygen species (ROS) such as superoxide anion and H2O2 that can attack numerous mitochondrial proteins including dihydrolipoamide dehydrogenase (DLDH). While DLDH is known to be vulnerable to oxidative inactivation, the mechanisms have not been clearly elucidated. The present study was therefore designed to investigate the mechanisms of DLDH oxidative inactivation by mitochondrial reactive oxygen species (ROS). Mitochondria, isolated from rat brain, were incubated with mitochondrial respiratory substrates such as pyruvate/malate or succinate in the presence of electron transport chain inhibitors such as rotenone or antimycin A. This is followed by enzyme activity assay and gel-based proteomic analysis. The present study also examined whether ROS-induced DLDH oxidative inactivation could be reversed by reducing reagents such as DTT, cysteine, and glutathione. Results show that DLDH could only be inactivated by complex III- but not complex I-derived ROS; and the accompanying loss of activity due to the inactivation could be restored by cysteine and glutathione, indicating that DLDH oxidative inactivation by complex III-derived ROS was a reversible process. Further studies using catalase indicate that it was H2O2 instead of superoxide anion that was responsible for DLDH inactivation. Moreover, using sulfenic acid-specific labeling techniques in conjunction with two-dimensional Western blot analysis, we show that protein sulfenic acid formation (also known as sulfenation) was associated with the loss of DLDH enzymatic activity observed under our experimental conditions. Additionally, such oxidative modification was shown to be associated with preventing DLDH from further inactivation by the thiol-reactive reagent N-ethylmaleimide. Taken together, the present study provides insights into the mechanisms of DLDH oxidative inactivation by mitochondrial H2O2.


Adaptive planning strategy for high dose rate prostate brachytherapy—a simulation study on needle positioning errors.
The development of magnetic resonance (MR) guided high dose rate (HDR) brachytherapy for prostate cancer has gained increasing interest for delivering a high tumor dose safely in a single fraction. To support needle placement in the limited workspace inside the closed-bore MRI, a single-needle MR-compatible robot is currently under development at the University Medical Center Utrecht (UMCU). This robotic device taps the needle in a divergent way from a single rotation point into the prostate. With this setup, it is warranted to deliver the irradiation dose by successive insertions of the needle. Although robot-assisted needle placement is expected to be more accurate than manual template-guided insertion, needle positioning errors may occur and are likely to modify the pre-planned dose distribution.In this paper, we propose a dose plan adaptation strategy for HDR prostate brachytherapy with feedback on the needle position: a dose plan is made at the beginning of the interventional procedure and updated after each needle insertion in order to compensate for possible needle positioning errors. The introduced procedure can be used with the single needle MR-compatible robot developed at the UMCU. The proposed feedback strategy was tested by simulating complete HDR procedures with and without feedback on eight patients with different numbers of needle insertions (varying from 4 to 12). In of the cases tested, the number of clinically acceptable plans obtained at the end of the procedure was larger with feedback compared to the situation without feedback. Furthermore, the computation time of the feedback between each insertion was below 100 s which makes it eligible for intra-operative use.


5-Hydroxytryptophan toxicosis in dogs: 21 cases (1989-1999).
To determine epidemiologic characteristics, clinical findings, and treatment outcome of 5-hydroxytryptophan (5-HTP) toxicosis in dogs. Retrospective study. 21 dogs with evidence of accidental 5-HTP ingestion. Information was retrieved from the National Animal Poison Control Center database. Records of dogs ingesting 5-HTP between January 1989 and February 1999 were reviewed for information on signalment, dose ingested, clinical signs (onset, severity, duration), treatments administered, and outcome. Clinical signs of toxicosis developed in 19 of 21 (90%) dogs. Neurologic signs included seizures (9 dogs), depression (6), tremors (5), hyperesthesia (5), and ataxia (4). Gastrointestinal tract signs included vomiting or diarrhea (12 dogs), signs of abdominal pain (3), and hypersalivation (2). Other clinical signs were hyperthermia (7 dogs) and transient blindness (3). Three dogs died. No important clinical laboratory or necropsy findings were reported. The doses of 5-HTP ingested ranged from 2.5 to 573 mg/kg (1.1 to 260 mg/lb) of body weight; the minimum toxic dose reported in our study was 23.6 mg/kg (10.7 mg/lb), and the minimum lethal dose was 128 mg/kg (58.1 mg/lb). Onset of signs ranged from 10 minutes to 4 hours after ingestion, and signs lasted up to 36 hours. Of 17 dogs with clinical signs of toxicosis that received treatment, 16 recovered; treatment consisted of decontamination, seizure control, thermoregulation, fluid therapy, and supportive care. Ingestion of 5-HTP in dogs can result in a potentially life-threatening syndrome resembling serotonin syndrome in humans, which requires prompt and aggressive care.


Effect of stepped care treatment on the incidence of myocardial infarction and angina pectoris. 5-Year findings of the hypertension detection and follow-up program.
Information on indices of coronary heart disease (CHD) and myocardial infarction (MI) (angina pectoris by Rose Questionnaire, MI by Rose Questionnaire, history, and electrocardiogram) was obtained in the Hypertension Detection and Follow-Up Program (U.S. National Institutes of Health) at baseline, Year 2, and Year 5 of follow-up. The presence of any of these findings at baseline markedly increased all-cause mortality during the 5 years of observation. In individuals with negative findings at baseline, the 5-year incidence of MI and angina pectoris by these indices was less in the Stepped Care than Referred Care cohort. These results are compatible with the conclusion that antihypertensive therapy reduces the incidence of symptomatic CHD.


Nicotinic acid- and monomethyl fumarate-induced flushing involves GPR109A expressed by keratinocytes and COX-2-dependent prostanoid formation in mice.
The antidyslipidemic drug nicotinic acid and the antipsoriatic drug monomethyl fumarate induce cutaneous flushing through activation of G protein-coupled receptor 109A (GPR109A). Flushing is a troublesome side effect of nicotinic acid, but may be a direct reflection of the wanted effects of monomethyl fumarate. Here we analyzed the mechanisms underlying GPR109A-mediated flushing and show that both Langerhans cells and keratinocytes express GPR109A in mice. Using cell ablation approaches and transgenic cell type-specific GPR109A expression in Gpr109a-/- mice, we have provided evidence that the early phase of flushing depends on GPR109A expressed on Langerhans cells, whereas the late phase is mediated by GPR109A expressed on keratinocytes. Interestingly, the first phase of flushing was blocked by a selective cyclooxygenase-1 (COX-1) inhibitor, and the late phase was sensitive to a selective COX-2 inhibitor. Both monomethyl fumarate and nicotinic acid induced PGE2 formation in isolated keratinocytes through activation of GPR109A and COX-2. Thus, the early and late phases of the GPR109A-mediated cutaneous flushing reaction involve different epidermal cell types and prostanoid-forming enzymes. These data will help to guide new efficient approaches to mitigate nicotinic acid-induced flushing and may help to exploit the potential antipsoriatic effects of GPR109A agonists in the skin.


Cardiovascular disease in hereditary haemophilia: The challenges of longevity.
The development of effective and safe treatments has significantly increased the life expectancy of persons with haemophilia (PWH). This has been accompanied by an increase in the comorbidities of ageing including cardiovascular disease, which poses particular challenges due to the opposing risks of bleeding from haemophilia and antithrombotic treatments versus thrombosis. Although mortality secondary to coronary artery disease in PWH is less than in the general population, the rate of atherosclerosis appears similar. The prevalence of atrial fibrillation in PWH and risk of secondary thromboembolic stroke are not well established. PWH can be safely supported through acute coronary interventions but data on the safety and efficacy of long-term antithrombotics are scarce. Increased awareness and research on cardiovascular disease in PWH will be crucial to improve primary prevention, acute management, secondary prevention and to best support ageing PWH.


Differential patterns of glucose-induced electrical activity and intracellular calcium responses in single mouse and rat pancreatic islets.
Although isolated rat islets are widely used to study in vitro insulin secretion and the underlying metabolic and ionic processes, knowledge on the properties of glucose-induced electrical activity (GIEA), a key step in glucose-response coupling, has been gathered almost exclusively from microdissected mouse islets. Using a modified intracellular recording technique, we have now compared the patterns of GIEA in collagenase-isolated rat and mouse islets. Resting membrane potentials of rat and mouse beta-cells were approximately -50 and -60 mV, respectively. Both rat and mouse beta-cells displayed prompt membrane depolarizations in response to glucose. However, whereas the latter exhibited a bursting pattern consisting of alternating hyperpolarized and depolarized active phases, rat beta-cells fired action potentials from a nonoscillating membrane potential at all glucose concentrations (8.4-22.0 mmol/l). This was mirrored by changes in the intracellular Ca2+ concentration ([Ca2+]i), which was oscillatory in mouse and nonoscillatory in rat islets. Stimulated rat beta-cells were strongly hyperpolarized by diazoxide, an activator of ATP-dependent K+ channels. Glucose evoked dose-dependent depolarizations and [Ca2+]i increases in both rat (EC50 5.9-6.9 mmol/l) and mouse islets (EC50 8.3-9.5 mmol/l), although it did not affect the burst plateau potential in the latter case. We conclude that there are important differences between beta-cells from both species with respect to early steps in the stimulus-secretion coupling cascade based on the following findings: 1) mouse beta-cells have a larger resting K+ conductance in 2 mmol/l glucose, 2) rat beta-cells lack the compensatory mechanism responsible for generating membrane potential oscillations and holding the depolarized plateau potential in mouse beta-cells, and 3) the electrical and [Ca2+]i dose-response curves in rat beta-cells are shifted toward lower glucose concentrations. Exploring the molecular basis of these differences may clarify several a priori assumptions on the electrophysiological properties of rat beta-cells, which could foster the development of new working models of pancreatic beta-cell function.


Auxin production is a common feature of most pathovars of Pseudomonas syringae.
We investigated indole-3-acetic acid (IAA) production by 57 pathovars of Pseudomonas syringae and related species. Most of those analyzed produced IAA, especially in the presence of tryptophan. Eight strains produced high IAA concentrations in the absence of Trp. The iaaM and iaaH genes of P. savastanoi pv. savastanoi were detected in a limited number of strains only, including the eight above-mentioned strains. Thus, IAA synthesis in most assayed strains of P. syringae and related species does not involve genes highly similar to iaaM and iaaH. In contrast, the iaaL gene encoding an IAA-lysine synthase was detected in most pathovars, and was often found on plasmids.


Use of Biomarkers to Guide Decisions on Adjuvant Systemic Therapy for Women With Early-Stage Invasive Breast Cancer: ASCO Clinical Practice Guideline Update-Integration of Results From TAILORx.
This focused update addresses the use of Oncotype DX in guiding decisions on the use of adjuvant systemic therapy. ASCO uses a signals approach to facilitate guideline updating. For this focused update, the publication of the Trial Assigning Individualized Options for Treatment (TAILORx) evaluating noninferiority of endocrine therapy alone versus chemoendocrine therapy for invasive disease-free survival in women with Oncotype DX scores provided a signal. An expert panel reviewed the results of TAILORx along with other published literature on the Oncotype DX assay to assess for evidence of clinical utility. For patients with hormone receptor-positive, axillary node-negative breast cancer whose tumors have Oncotype DX recurrence scores of less than 26, there is little to no benefit from chemotherapy, especially for patients older than age 50 years. Clinicians may recommend endocrine therapy alone for women older than age 50 years. For patients 50 years of age or younger with recurrence scores of 16 to 25, clinicians may offer chemoendocrine therapy. Patients with recurrence scores greater than 30 should be considered candidates for chemoendocrine therapy. Based on informal consensus, the panel recommends that oncologists may offer chemoendocrine therapy to these patients with recurrence scores of 26 to 30. Additional information can be found at www.asco.org/breast-cancer-guidelines.


Quality of life in preoperative patients with sacroiliac joint dysfunction is at least as depressed as in other lumbar spinal conditions.
Pain from the sacroiliac joint (SIJ) is an under-recognized cause of low back pain. The degree to which SIJ pain decreases quality of life has not been directly compared to other more familiar conditions of the lumbar spine. Multivariate regression analysis of individual patient data from two prospective multicenter clinical trials of SIJ fusion and three prospective multicenter clinical trials of surgical treatments for degenerative lumbar spine conditions. Controlling for baseline demographic parameters as well as a validated disability score, quality of life scores (EuroQOL 5-D and SF-36) were, in most cases, lower in the SIJ cohorts compared to the three other spine surgery cohorts. Patients with SIJ dysfunction considering surgery have decrements in quality of life as or more severe compared to patients with degenerative spondylolisthesis, spinal stenosis, and intervertebral disc herniation.


Economic evaluations of pharmacist-managed services in people with diabetes mellitus: a systematic review.
To review and evaluate the most recent literature on the economic outcomes of pharmacist-managed services in people with diabetes. The global prevalence of diabetes is increasing. Although pharmacist-managed services have been shown to improve people's health outcomes, the economic impact of these programmes remains unclear. A systematic review was conducted of six databases. Study inclusion criteria were: (1) original research; (2) evaluation of pharmacist-managed services in people with diabetes; (3) an economic evaluation; (4) English-language publication; and (5) full-text, published between January 2006 and December 2014. The quality of the full economic evaluations reviewed was evaluated using the Consolidated Health Economic Evaluation Reporting Standards checklist. A total of 2204 articles were screened and 25 studies were selected. These studies were conducted in a community pharmacy (n = 10), a clinic- /hospital-based outpatient facility (n = 8), or others. Pharmacist-managed services included targeted education (n = 24), general pharmacotherapeutic monitoring (n = 21), health screening or laboratory testing services (n = 9), immunization services (n = 2) and pharmacokinetic monitoring (n = 1). Compared with usual care, pharmacist-managed services resulted in cost savings that varied from $7 to $65,000 ($8 to $85,000 in 2014 US dollars) per person per year, and generated higher quality-adjusted life years with lower costs. Benefit-to-cost ratios ranged from 1:1 to 8.5:1. Among the 25 studies reviewed, 11 were full economic evaluations of moderate quality. Pharmacist-managed services had a positive return in terms of economic viability. With the expanding role of pharmacists in the healthcare sector, alongside increasing health expenditure, future economic studies of high quality are needed to investigate the cost-effectiveness of these services.


Local anaesthetics and regional anaesthesia for preventing chronic pain after surgery.
Regional anaesthesia may reduce the rate of persistent (chronic) pain after surgery, a frequent and debilitating condition. To compare local anaesthetics and regional anaesthesia versus conventional analgesia for the prevention of persistent pain six or 12 months after surgery. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2012, Issue 4), PubMed (1966 to April 2012), EMBASE (1966 to May 2012) and CINAHL (1966 to May 2012) without any language restriction. We used a combination of free text search and controlled vocabulary search. The results were limited to randomized controlled clinical trials (RCTs). We conducted a handsearch in reference lists of included trials, review articles and conference abstracts. We included RCTs comparing local anaesthetics or regional anaesthesia versus conventional analgesia with a pain outcome at six or 12 months after surgery. Two authors independently assessed trial quality and extracted data, including information on adverse events. We contacted study authors for additional information. Results are presented as pooled odds ratios (OR) with 95% confidence intervals (CI), based on random-effects models (inverse variance method). We grouped studies according to surgical interventions. We employed the Chi(2) test and calculated the I(2) statistic to investigate study heterogeneity. We identified 23 RCTs studying local anaesthetics or regional anaesthesia for the prevention of persistent (chronic) pain after surgery. Data from a total of 1090 patients with outcomes at six months and of 441 patients with outcomes at 12 months were presented. No study included children. We pooled data from 250 participants after thoracotomy, with outcomes at six months. Data favoured regional anaesthesia for the prevention of chronic pain at six months after thoracotomy with an OR of 0.33 (95% CI 0.20 to 0.56). We pooled two studies on paravertebral block for breast cancer surgery; the pooled data of 89 participants with outcomes at five to six months favoured paravertebral block with an OR of 0.37 (95% CI 0.14 to 0.94).The methodological quality of the included studies was intermediate. Adverse effects were not studied systematically and were reported sparsely. Clinical heterogeneity, attrition and sparse outcome data hampered the assessment of effects, especially at 12 months. Epidural anaesthesia may reduce the risk of developing chronic pain after thoracotomy in about one patient out of every four patients treated. Paravertebral block may reduce the risk of chronic pain after breast cancer surgery in about one out of every five women treated. Our conclusions are significantly weakened by performance bias, shortcomings in allocation concealment, considerable attrition and incomplete outcome data. We caution that our evidence synthesis is based on only a few, small studies. More studies with high methodological quality, addressing various types of surgery and different age groups, including children, are needed.


Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
The actin cytoskeleton of dendritic spines plays a key role in morphological aspects of synaptic plasticity. The detailed analysis of the spine structure and dynamics in live neurons, however, has been hampered by the diffraction-limited resolution of conventional fluorescence microscopy. The advent of nanoscopic imaging techniques thus holds great promise for the study of these processes. We implemented a strategy for the visualization of morphological changes of dendritic spines over tens of minutes at a lateral resolution of 25 to 65 nm. We have generated a low-affinity photoconvertible probe, capable of reversibly binding to actin and thus allowing long-term photoactivated localization microscopy of the spine cytoskeleton. Using this approach, we resolve structural parameters of spines and record their long-term dynamics at a temporal resolution below one minute. Furthermore, we have determined changes in the spine morphology in response to pharmacologically induced synaptic activity and quantified the actin redistribution underlying these changes. By combining PALM imaging with quantum dot tracking, we could also simultaneously visualize the cytoskeleton and the spine membrane, allowing us to record complementary information on the morphological changes of the spines at super-resolution.


Understanding spatial connectivity of individuals with non-uniform population density.
We construct a two-dimensional geometric graph connecting individuals placed in space within a given contact distance. The individuals are distributed using a measured country's density of population. We observe that while large clusters (group of individuals connected) emerge within some regions, they are trapped in detached urban areas owing to the low population density of the regions bordering them. To understand the emergence of a giant cluster that connects the entire population, we compare the empirical geometric graph with the one generated by placing the same number of individuals randomly in space. We find that, for small contact distances, the empirical distribution of population dominates the growth of connected components, but no critical percolation transition is observed in contrast to the graph generated by a random distribution of population. Our results show that contact distances from real-world situations as for WIFI and Bluetooth connections drop in a zone where a fully connected cluster is not observed, hinting that human mobility must play a crucial role in contact-based diseases and wireless viruses' large-scale spreading.


Bacterial communities associated with four ctenophore genera from the German Bight (North Sea).
Intense research has been conducted on jellyfish and ctenophores in recent years. They are increasingly recognized as key elements in the marine ecosystem that serve as critical indicators and drivers of ecosystem performance and change. However, the bacterial community associated with ctenophores is still poorly investigated. Based on automated ribosomal intergenic spacer analysis (ARISA) and 16S ribosomal RNA gene amplicon pyrosequencing, we investigated bacterial communities associated with the frequently occurring ctenophore species Mnemiopsis leidyi, Beroe sp., Bolinopsis infundibulum and Pleurobrachia pileus at Helgoland Roads in the German Bight (North Sea). We observed significant differences between the associated bacterial communities of the different ctenophore species based on ARISA patterns. With respect to bacterial taxa, all ctenophore species were dominated by Proteobacteria as revealed by pyrosequencing. Mnemiopsis leidyi and P. pileus mainly harboured Gammaproteobacteria, with Marinomonas as the dominant phylotype of M. leidyi. By contrast, Pseudoalteromonas and Psychrobacter were the most abundant Gammaproteobacteria in P. pileus. Beroe sp. was mainly dominated by Alphaproteobacteria, particularly by the genus Thalassospira. For B. infundibulum, the bacterial community was composed of Alphaproteobacteria and Gammaproteobacteria in equal parts, which consisted of the genera Thalassospira and Marinomonas. In addition, the bacterial communities associated with M. leidyi display a clear variation over time that needs further investigation. Our results indicate that the bacterial communities associated with ctenophores are highly species- specific.


An fMRI study exploring the overlap and differences between neural representations of physical and recalled pain.
Implementing a recall paradigm without hypnosis, we use functional MRI (fMRI) to explore and compare nociceptive and centrally-driven pain experiences. We posit that a trace of a recent nociceptive event can be used to create sensory-re-experiencing of pain that can be qualified in terms of intensity and vividness. Fifteen healthy volunteers received three levels of thermal stimuli (warm, low pain and high pain) and subsequently were asked to recall and then rate this experience. Neuroimaging results reveal that recalling a previous sensory experience activates an extensive network of classical pain processing structures except the contralateral posterior insular cortex. Nociceptive-specific activation of this structure and the rated intensity difference between physical and recalled pain events allow us to investigate the link between the quality of the original nociceptive stimulus and the mental trace, as well as the differences between the accompanying neural responses. Additionally, by incorporating the behavioural ratings, we explored which brain regions were separately responsible for generating either an accurate or vivid recall of the physical experience. Together, these observations further our understanding of centrally-mediated pain experiences and pain memory as well as the potential relevance of these factors in the maintenance of chronic pain.


Actively Targeted Nanoparticles for Drug Delivery to Tumor.
Nanomedicine is an emerging therapeutic modality. Nanoparticles (NPs) are potential vehicles for delivery of anticancer therapeutics. NPs can be designed to facilitate tumor drug delivery both by passive and active targeting mechanisms. Passive targeting of NPs to tumors can be achieved through the enhanced permeability and retention (EPR) effect. Meanwhile, actively targeted NPs can be designed based on two different targeting mechanisms, ligand-directed targeting to the tumor cells and tumor microenvironment (TME)-directed targeting. We searched for and reviewed recently published literature on actively targeted NPs. Progress in this field was summarized in several focus areas, including methods for targeting of tumor cells and for targeting TME. Advantages and limitations of each approach were discussed. This article covers data from 240 recent publications and provided numerous examples of ligand-directed NPs targeting tumor cell-selective surface receptors. Targeting ligands discussed include proteins such as transferrin and antibodies, as well as low molecular weight agents, such as peptides, aptamers, carbohydrates, and folate. In addition, extensive discussions of TME targeting NPs, designed to release drug in response to TME-specific stimuli, such as low pH, tumor-selective enzymes, and unique characteristics of tumor neovasculature, are also included in this review. In general, many novel actively targeting strategies have been developed and encouraging data have been reported in numerous settings, both in vitro and in animal studies. Active targeting of NPs has experienced rapid growth as a field of research and is continuously expanding. There are now some early examples of efforts on clinical translation and reported clinical trials on these NPs. Future development of actively targeted NPs depends on better understanding of the many factors affecting the behavior of NPs in vivo and likely involves combining the approaches of targeting the tumor cells and of targeting components of the TME.


Structure of the Lassa virus glycan shield provides a model for immunological resistance.
Lassa virus is an Old World arenavirus endemic to West Africa that causes severe hemorrhagic fever. Vaccine development has focused on the envelope glycoprotein complex (GPC) that extends from the virion envelope. The often inadequate antibody immune response elicited by both vaccine and natural infection has been, in part, attributed to the abundance of N-linked glycosylation on the GPC. Here, using a virus-like-particle system that presents Lassa virus GPC in a native-like context, we determine the composite population of each of the N-linked glycosylation sites presented on the trimeric GPC spike. Our analysis reveals the presence of underprocessed oligomannose-type glycans, which form punctuated clusters that obscure the proteinous surface of both the GP1 attachment and GP2 fusion glycoprotein subunits of the Lassa virus GPC. These oligomannose clusters are seemingly derived as a result of sterically reduced accessibility to glycan processing enzymes, and limited amino acid diversification around these sites supports their role protecting against the humoral immune response. Combined, our data provide a structure-based blueprint for understanding how glycans render the glycoprotein spikes of Lassa virus and other Old World arenaviruses immunologically resistant targets.


Accounting for risk in valuing forest carbon offsets.
Forests can sequester carbon dioxide, thereby reducing atmospheric concentrations and slowing global warming. In the U.S., forest carbon stocks have increased as a result of regrowth following land abandonment and in-growth due to fire suppression, and they currently sequester approximately 10% of annual US emissions. This ecosystem service is recognized in greenhouse gas protocols and cap-and-trade mechanisms, yet forest carbon is valued equally regardless of forest type, an approach that fails to account for risk of carbon loss from disturbance. Here we show that incorporating wildfire risk reduces the value of forest carbon depending on the location and condition of the forest. There is a general trend of decreasing risk-scaled forest carbon value moving from the northern toward the southern continental U.S. Because disturbance is a major ecological factor influencing long-term carbon storage and is often sensitive to human management, carbon trading mechanisms should account for the reduction in value associated with disturbance risk.


A comparison of Peritest automated perimetry and Goldmann perimetry.
Both Peritest perimetry and Goldmann visual field testing were performed on three groups of eyes. Eighty-one eyes had glaucoma with elevated intraocular pressures and abnormal visual fields determined by Goldmann perimetry. There were 47 eyes with suspected glaucoma, based on elevated IOPs and normal Goldmann visual fields. Nineteen eyes were normal. All visual fields were evaluated in masked fashion, and the results were compared. The Peritest had a high sensitivity in both glaucomatous (98.8%) and normal (94.7%) eyes. One half of the eyes with suspected glaucoma had abnormal Peritest results but normal Goldmann visual fields. These eyes were examined prospectively with color stereo discs and monochromatic nerve fiber layer photographs. One half of the eyes with suspected glaucoma and abnormal Peritest results also had photographic evidence suggesting optic nerve damage. Thus, early field defects detected by the Peritest perimeter in eyes with suspected glaucoma are likely to be representative of glaucomatous damage.


Time course of stretch-induced isometric strength deficits.
Stretching in the warm-up has been shown to decrease several muscular performance variables, but the dose-response of this effect is unknown. This study documented the change in isometric grip strength over ten trials in a convenience sample of young adults randomly assigned to control (n=22) or repeated bouts of 10-s static stretches of the wrist flexors (n=35). There was a significant (P<0.05) difference in the change in mean normalized grip strength between the control and stretching groups that was not significantly different across gender. Grip strengths in the control group were consistent with a linear trend, while the grip strengths in the stretching group declined in a logarithmic fashion to 88.8% with 100 s of stretching. Statistically significant (P<0.05) differences in normalized grip strength between the two groups appeared after 40 s of stretching. Meaningful decreases in isometric grip strength following static stretching are likely to appear in young adults following 20-40 s of static stretching.


Safety and Accuracy of Minimally Invasive Long Bone Fracture Repair Using a 2.5-mm Interlocking Nail: A Cadaveric Feline Study.
The Targon Vet System (TVS) is a 2.5-mm interlocking nail that can be applied minimally invasively. The purpose of this study was to test if the TVS could be safely applied percutaneously to different feline long bones without fluoroscopic guidance. A gap fracture was created in 96 feline humeri, femora and tibiae (n = 32/group). Paired bones were randomly assigned to two treatment groups: (1) TVS inserted percutaneously with fluoroscopy and (2) TVS inserted percutaneously without fluoroscopy. Intraoperative evaluation (complications, procedure time, attempts), radiographs (pre-/postoperative alignment, length) and anatomical dissection (neurovascular injury, rotational alignment) were compared between treatment groups. The use of fluoroscopy did not lead to significant differences in any of the outcome measures. Intraoperative complications predominantly occurred in the distal humerus (12/32) and the proximal femur (7/32). In total, 20/96 complications occurred with no complications for the tibia. Neurovascular structures were only damaged at the medial side of the distal humerus (10/32). We conclude that the TVS can be safely applied percutaneously to the tibia and with limitations to the femur in normal cadaveric cats without fluoroscopy. Despite the limitations of a cadaveric study, the high number of complications is leading us to consider the humerus not safe for the TVS. A learning curve has to be expected and technical recommendations should be respected to decrease complications.


Molecular similarity between myelodysplastic form of chronic myelomonocytic leukemia and refractory anemia with ring sideroblasts.
Chronic myelomonocytic leukemia is similar to but a separate entity from both myeloproliferative neoplasms and myelodysplastic syndromes, and shows either myeloproliferative or myelodysplastic features. We ask whether this distinction may have a molecular basis. We established the gene expression profiles of 39 samples of chronic myelomonocytic leukemia (including 12 CD34-positive) and 32 CD34-positive samples of myelodysplastic syndromes by using Affymetrix microarrays, and studied the status of 18 genes by Sanger sequencing and array-comparative genomic hybridization in 53 samples. Analysis of 12 mRNAS from chronic myelomonocytic leukemia established a gene expression signature of 122 probe sets differentially expressed between proliferative and dysplastic cases of chronic myelomonocytic leukemia. As compared to proliferative cases, dysplastic cases over-expressed genes involved in red blood cell biology. When applied to 32 myelodysplastic syndromes, this gene expression signature was able to discriminate refractory anemias with ring sideroblasts from refractory anemias with excess of blasts. By comparing mRNAS from these two forms of myelodysplastic syndromes we derived a second gene expression signature. This signature separated the myelodysplastic and myeloproliferative forms of chronic myelomonocytic leukemias. These results were validated using two independent gene expression data sets. We found that myelodysplastic chronic myelomonocytic leukemias are characterized by mutations in transcription/epigenetic regulators (ASXL1, RUNX1, TET2) and splicing genes (SRSF2) and the absence of mutations in signaling genes. Myelodysplastic chronic myelomonocytic leukemias and refractory anemias with ring sideroblasts share a common expression program suggesting they are part of a continuum, which is not totally explained by their similar but not, however, identical mutation spectrum.


The expression of 70 apoptosis genes in relation to lineage, genetic subtype, cellular drug resistance, and outcome in childhood acute lymphoblastic leukemia.
Childhood acute lymphoblastic leukemia (ALL) consists of various subtypes that respond differently to cytotoxic drugs and therefore have a markedly different clinical outcome. We used microarrays to investigate, in 190 children with ALL at initial diagnosis, whether 70 key apoptosis genes were differentially expressed between leukemic subgroups defined by lineage, genetic subtype, in vitro drug resistance, and clinical outcome. The expression of 44 of 70 genes was significantly different in T-versus B-lineage ALL, 22 genes differed in hyperdiploid versus nonhyperdiploid, 16 in TEL-AML1-positive versus-negative, and 13 in E2A-rearranged versus germ-line B-lineage ALL. Expression of MCL1 and DAPK1 was significantly associated with prednisolone sensitivity, whereas BCL2L13, HRK, and TNF were related to L-asparaginase resistance. BCL2L13 overexpression was also associated with unfavorable clinical outcome (P < .001). Multivariate analysis including known risk factors revealed that BCL2L13 expression was an independent prognostic factor (P = .011). The same trend was observed in a validation group of 92 children with ALL treated on a different protocol at St Jude (P = .051). In conclusion, ALL subtypes have a unique expression pattern of apoptosis genes and our data suggest that selective genes are linked to cellular drug resistance and prognosis in childhood B-lineage ALL.


Clinical monitoring scales in acute brain injury: assessment of coma, pain, agitation, and delirium.
Serial clinical examination represents the most fundamental and basic form of neurological monitoring, and is often the first and only form of such monitoring in patients. Even in patients subjected to physiological monitoring using a range of technologies, the clinical examination remains an essential tool to follow neurological progress. Key aspects of the clinical examination have now been systematized into scoring schemes, and address consciousness, pain, agitation, and delirium (PAD). The Glasgow Coma Scale has been the traditional tool to measure consciousness, but the full outline of unresponsiveness (FOUR) score has recently been validated in a variety of settings, and at present, both represent clinically useful tools. Assessment of PAD in neurologically compromised patients present special challenges. For pain, the Numeric Rating Scale is the preferred initial approach, with either the Behavioral Pain Scale or the Critical Care Pain Observation Tool in subjects who are not able to respond. The Nociception Coma Scale-Revised may be useful in patients with severe disorders of consciousness. Conventional sedation scoring tools for critical care, such as the Richmond Area Sedation Scale (RASS) and Sedation-Agitation Scale (SAS) may provide reasonable tools in some neurocritical care patients. The use of sedative drugs and neuromuscular blockers may invalidate the use of some clinical examination tools in others. The use of sedation interruption to assess neurological status can result in physiological derangement in unstable patients (such as those with uncontrolled intracranial hypertension), and is not recommended.


Time lines of infection and disease in human influenza: a review of volunteer challenge studies.
The dynamics of viral shedding and symptoms following influenza virus infection are key factors when considering epidemic control measures. The authors reviewed published studies describing the course of influenza virus infection in placebo-treated and untreated volunteers challenged with wild-type influenza virus. A total of 56 different studies with 1,280 healthy participants were considered. Viral shedding increased sharply between 0.5 and 1 day after challenge and consistently peaked on day 2. The duration of viral shedding averaged over 375 participants was 4.80 days (95% confidence interval: 4.31, 5.29). The frequency of symptomatic infection was 66.9% (95% confidence interval: 58.3, 74.5). Fever was observed in 37.0% of A/H1N1, 40.6% of A/H3N2 (p = 0.86), and 7.5% of B infections (p = 0.001). The total symptoms scores increased on day 1 and peaked on day 3. Systemic symptoms peaked on day 2. No such data exist for children or elderly subjects, but epidemiologic studies suggest that the natural history might differ. The present analysis confirms prior expert opinion on the duration of viral shedding or the frequency of asymptomatic influenza infection, extends prior knowledge on the dynamics of viral shedding and symptoms, and provides original results on the frequency of respiratory symptoms or fever.


Relative effects of aging and age-related macular degeneration on peripheral visual function.
The purpose of this study was to determine the extent of peripheral visual deficits in patients with early age-related macular degeneration (ARMD) using electrophysiological and psychophysical techniques. Dark-adaptation curves, electro-oculograms (EOGs), and electroretinograms (ERGs) were obtained from patients with early ARMD and from normally sighted control subjects. The control subjects' data were used to calculate age-dependent 95% confidence intervals for each measure of visual function. For the control subjects, performance on all our measures of visual function decreased with age. For the patients with early ARMD, the cone system absolute thresholds, EOG ratios, and cone-dominated ERG amplitudes and implicit times were within the range of normal age-related changes. Rod system absolute thresholds, cone-rod break times, and rod-dominated electroretinographic measures were abnormal in some patients. These results suggest that when the effects of aging are taken into account, some patients classified as early ARMD may not show significant changes in peripheral retinal function with standard clinical tests.


Secondhand smoke exposure and mental health among children and adolescents.
To examine a potential association between biologically confirmed secondhand smoke exposure and symptoms of Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) (DSM-IV) major depressive disorder, generalized anxiety disorder, panic disorder, attention-deficit/hyperactivity disorder, and conduct disorder using a nationally representative sample of US children and adolescents. Nationally representative cross-sectional survey of the United States. Continental United States. Children and adolescents aged 8 to 15 years who participated in the National Health and Nutrition Examination Survey from 2001 to 2004. Measurement of serum cotinine level to assess secondhand smoke exposure among nonsmokers. The DSM-IV symptoms were derived from selected modules of the National Institute of Mental Health's Diagnostic Interview Schedule for Children Version IV, a structured diagnostic interview administered by trained lay interviewers. Among nonsmokers, serum cotinine level was positively associated with symptoms of DSM-IV major depressive disorder, generalized anxiety disorder, attention-deficit/hyperactivity disorder, and conduct disorder after adjusting for survey design, age, sex, race/ethnicity, poverty, migraine, asthma, hay fever, maternal smoking during pregnancy, and allostatic load. Associations with serum cotinine level were more apparent for boys and for participants of non-Hispanic white race/ethnicity. Our results are consistent with a growing body of research documenting an association between secondhand smoke exposure and mental health outcomes. Future research is warranted to establish the biological or psychological mechanisms of association.


Mutant versions of the S. cerevisiae transcription elongation factor Spt16 define regions of Spt16 that functionally interact with histone H3.
In eukaryotic cells, the highly conserved FACT (FAcilitates Chromatin Transcription) complex plays important roles in several chromatin-based processes including transcription initiation and elongation. During transcription elongation, the FACT complex interacts directly with nucleosomes to facilitate histone removal upon RNA polymerase II (Pol II) passage and assists in the reconstitution of nucleosomes following Pol II passage. Although the contribution of the FACT complex to the process of transcription elongation has been well established, the mechanisms that govern interactions between FACT and chromatin still remain to be fully elucidated. Using the budding yeast Saccharomyces cerevisiae as a model system, we provide evidence that the middle domain of the FACT subunit Spt16--the Spt16-M domain--is involved in functional interactions with histone H3. Our results show that the Spt16-M domain plays a role in the prevention of cryptic intragenic transcription during transcription elongation and also suggest that the Spt16-M domain has a function in regulating dissociation of Spt16 from chromatin at the end of the transcription process. We also provide evidence for a role for the extreme carboxy terminus of Spt16 in functional interactions with histone H3. Taken together, our studies point to previously undescribed roles for the Spt16 M-domain and extreme carboxy terminus in regulating interactions between Spt16 and chromatin during the process of transcription elongation.


Alternative splicing of MDM2 mRNA in lung carcinomas and lung cell lines.
The MDM2 gene is overexpressed in several human tumors and its product may be processed into various isoforms. Recently, alternative splicing forms of MDM2 mRNA have been detected in various types of tumors. In this study, lung tissue from human non small cell lung cancers was examined for MDM2 mRNA splicing variants by nested RT-PCR. Of the 117 lung cancer tissue samples analyzed, a total of 31 (26.5%) had splice variants for the MDM2 gene, while 59 (50.4%) had undetectable levels of MDM2 transcript. Further analysis indicated that the predominant variant for 26 of the 31 samples with alternative MDM2 splicing products was MDM2-657, a splice variant lacking exons 3-11. Significant associations were found between the frequency of alternative splicing and the gender and smoking habits of the patients. Approximately 36% of male patients had alternative splicing of MDM2 compared with only 9.5% of female patients (P = 0.008); 44.2% of the smoker patients had alternative MDM2 splice forms versus 16.2% of nonsmokers (P = 0.003). Furthermore, most normal lung cell lines examined possessed only full-length MDM2 mRNA, while among several lung cancer cell lines, only H1355 and CaLu-1 cells lacked alternatively spliced MDM2 transcripts. When H1355 cells were treated in vitro with the cigarette smoke carcinogen benzo[a]pyrene (B[a]P) or the B[a]P metabolite benzo[a]pyrene diolepoxide (BPDE), three MDM2 splicing products were detected by nested RT-PCR. Finally, with the use of several specific inhibitors, we found that BPDE-induced MDM2 mRNA alternative splicing in H1355 cells may occur through the PI3K or MAPK pathway. Overall, our results suggest that carcinogens present in cigarette smoke increase the risk of alternative MDM2 splicing, which is highly associated with lung cancer.