Impact of chronic obstructive pulmonary disease on survival and symptoms of severe aortic valve stenosis.
As patients with severe aortic valve stenosis (AS) develop symptoms their survival decreases rapidly, if treated conservatively. Transcatheter aortic valve implantation (TAVI) has been introduced as a less invasive treatment alternative, especially in inoperable patients, who often have severe comorbidities, including chronic obstructive pulmonary disease (COPD). Since the beginning of our TAVI program in March 2008, data on all 131 TAVI patients were prospectively and consecutively collected in this registry with complete follow-up. COPD was present in 37 patients. By January 2012 survival data were collected from the Danish Civil Registration System. Median follow-up duration was 559 days. Overall survival and survival from cardiac death was equivalent in both patients with and without COPD (p = 0.98 and p = 0.26) in the follow-up period. Further, patients with COPD had higher New York Heart Association (NYHA) class prior to intervention compared with those without (3.1 ± 0.5 vs. 2.9 ± 0.5, p = 0.02). In multivariate regression analysis COPD was associated with 30-day postoperative NYHA class (0.43; 95% confidence interval (CI): 0.10-0.75; p = 0.01), but not to NYHA class improvement from pre- to postintervention (0.25; 95% CI: - 0.12 to -0.63; p = 0.18). In patients with symptomatic severe AS treated with TAVI, the presence of COPD neither affects overall survival nor survival from cardiac death. Patients with COPD had, however, both higher pre- and postoperative NYHA class compared with patients without COPD, but NYHA class improvement from pre- to postintervention was equivalent in both groups.


High Summer Temperatures and Mortality in Estonia.
On-going climate change is predicted to result in a growing number of extreme weather events-such as heat waves-throughout Europe. The effect of high temperatures and heat waves are already having an important impact on public health in terms of increased mortality, but studies from an Estonian setting are almost entirely missing. We investigated mortality in relation to high summer temperatures and the time course of mortality in a coastal and inland region of Estonia. We collected daily mortality data and daily maximum temperature for a coastal and an inland region of Estonia. We applied a distributed lag non-linear model to investigate heat related mortality and the time course of mortality in Estonia. We found an immediate increase in mortality associated with temperatures exceeding the 75th percentile of summer maximum temperatures, corresponding to approximately 23°C. This increase lasted for a couple of days in both regions. The total effect of elevated temperatures was not lessened by significant mortality displacement. We observed significantly increased mortality in Estonia, both on a country level as well as for a coastal region and an inland region with a more continental climate. Heat related mortality was higher in the inland region as compared to the coastal region, however, no statistically significant differences were observed. The lower risks in coastal areas could be due to lower maximum temperatures and cooling effects of the sea, but also better socioeconomic condition. Our results suggest that region specific estimates of the impacts of temperature extremes on mortality are needed.


Cell matrix adhesion in cell migration.
The ability of cells to migrate is a fundamental physiological process involved in embryonic development, tissue homeostasis, immune surveillance and wound healing. In order for cells to migrate, they must interact with their environment using adhesion receptors, such as integrins, and form specialized adhesion complexes that mediate responses to different extracellular cues. In this review, we discuss the role of integrin adhesion complexes (IACs) in cell migration, highlighting the layers of regulation that are involved, including intracellular signalling cascades, mechanosensing and reciprocal feedback to the extracellular environment. We also discuss the role of IACs in extracellular matrix remodeling and how they impact upon cell migration.


Nanoparticle shape anisotropy and photoluminescence properties: Europium containing ZnO as a Model Case.
The precise control over electronic and optical properties of semiconductor (SC) materials is pivotal for a number of important applications like in optoelectronics, photocatalysis or in medicine. It is well known that the incorporation of heteroelements (doping as a classical case) is a powerful method for adjusting and enhancing the functionality of semiconductors. Independent from that, there already has been a tremendous progress regarding the synthesis of differently sized and shaped SC nanoparticles, and quantum-size effects are well documented experimentally and theoretically. Whereas size and shape control of nanoparticles work fairly well for the pure compounds, the presence of a heteroelement is problematic because the impurities interfere strongly with bottom up approaches applied for the synthesis of such particles, and effects are even stronger, when the heteroelement is aimed to be incorporated into the target lattice for chemical doping. Therefore, realizing coincident shape control of nanoparticle colloids and their doping still pose major difficulties. Due to a special mechanism of the emulsion based synthesis method presented here, involving a gelation of emulsion droplets prior to crystallization of shape-anisotropic ZnO nanoparticles, heteroelements can be effectively entrapped inside the lattice. Different nanocrystal shapes such as nanorods, -prisms, -plates, and -spheres can be obtained, determined by the use of certain emulsification agents. The degree of morphologic alterations depends on the type of incorporated heteroelement M(n+), concentration, and it seems that some shapes are more tolerant against doping than others. Focus was then set on the incorporation of Eu(3+) inside the ZnO particles, and it was shown that nanocrystal shape and aspect ratios could be adjusted while maintaining a fixed dopant level. Special PL properties could be observed implying energy transfer from ZnO excited near its band-gap (3.3 eV) to the Eu(3+) states mediated by defect luminescence of the nanoparticles. Indications for an influence of shape on photoluminescence (PL) properties were found. Finally, rod-like Eu@ZnO colloids were used as tracers to investigate their uptake into biological samples like HeLa cells. The PL was sufficient for identifying green and red emission under visible light excitation.


Lung inflammation persists after 27 hours of protective Acute Respiratory Distress Syndrome Network Strategy and is concentrated in the nondependent lung.
PET with [18F]fluoro-2-deoxy-D-glucose can be used to image cellular metabolism, which during lung inflammation mainly reflects neutrophil activity, allowing the study of regional lung inflammation in vivo. We aimed at studying the location and evolution of inflammation by PET imaging, relating it to morphology (CT), during the first 27 hours of application of protective-ventilation strategy as suggested by the Acute Respiratory Distress Syndrome Network, in a porcine experimental model of acute respiratory distress syndrome. Prospective laboratory investigation. University animal research laboratory. Ten piglets submitted to an experimental model of acute respiratory distress syndrome. Lung injury was induced by lung lavages and 210 minutes of injurious mechanical ventilation using low positive end-expiratory pressure and high inspiratory pressures. During 27 hours of controlled mechanical ventilation according to Acute Respiratory Distress Syndrome Network strategy, the animals were studied with dynamic PET imaging of [18F]fluoro-2-deoxy-D-glucose at two occasions with 24-hour interval between them. [18F]fluoro-2-deoxy-D-glucose uptake rate was computed for the total lung, four horizontal regions from top to bottom (nondependent to dependent regions) and for voxels grouped by similar density using standard Hounsfield units classification. The global lung uptake was elevated at 3 and 27 hours, suggesting persisting inflammation. In both PET acquisitions, nondependent regions presented the highest uptake (p = 0.002 and p = 0.006). Furthermore, from 3 to 27 hours, there was a change in the distribution of regional uptake (p = 0.003), with more pronounced concentration of inflammation in nondependent regions. Additionally, the poorly aerated tissue presented the largest uptake concentration after 27 hours. Protective Acute Respiratory Distress Syndrome Network strategy did not attenuate global pulmonary inflammation during the first 27 hours after severe lung insult. The strategy led to a concentration of inflammatory activity in the upper lung regions and in the poorly aerated lung regions. The present findings suggest that the poorly aerated lung tissue is an important target of the perpetuation of the inflammatory process occurring during ventilation according to the Acute Respiratory Distress Syndrome Network strategy.


Jasmonate-Responsive ERF Transcription Factors Regulate Steroidal Glycoalkaloid Biosynthesis in Tomato.
Steroidal glycoalkaloids (SGAs) are cholesterol-derived specialized metabolites produced in species of the Solanaceae. Here, we report that a group of jasmonate-responsive transcription factors of the ETHYLENE RESPONSE FACTOR (ERF) family (JREs) are close homologs of alkaloid regulators in Cathranthus roseus and tobacco, and regulate production of SGAs in tomato. In transgenic tomato, overexpression and dominant suppression of JRE genes caused drastic changes in SGA accumulation and in the expression of genes for metabolic enzymes involved in the multistep pathway leading to SGA biosynthesis, including the upstream mevalonate pathway. Transactivation and DNA-protein binding assays demonstrate that JRE4 activates the transcription of SGA biosynthetic genes by binding to GCC box-like elements in their promoters. These JRE-binding elements occur at significantly higher frequencies in proximal promoter regions of the genes regulated by JRE genes, supporting the conclusion that JREs mediate transcriptional co-ordination of a series of metabolic genes involved in SGA biosynthesis.


Role of Salt and Water in the Plasticization of PDAC/PSS Polyelectrolyte Assemblies.
In this work, we investigate the effect of salt and water on plasticization and thermal properties of hydrated poly(diallyldimethylammonium chloride) (PDAC) and poly(sodium 4-styrenesulfonate) (PSS) assemblies via molecular dynamics simulations and modulated differential scanning calorimetry (MDSC). Commonly, both water and salt are considered to be plasticizers of hydrated polyelectrolyte assemblies. However, the simulation results presented here show that while water has a plasticizing effect, salt can also have an opposite effect on the PE assemblies. On one hand, the presence of salt ions provides additional free volume for chain motion and weakens PDAC-PSS ion pairing due to electrostatic screening, which contributes toward plasticization of the complex. On the other hand, salt ions bind water in their hydration shells, which decreases water mobility and reduces the plasticization by hydration. Our MDSC results connect the findings to macroscopic PE plasticization and the glass-transition-like thermal transition Ttr under controlled PE hydration and salt content. This work identifies and characterizes the dual nature of salt both as plasticizer and hardener of PE assemblies and maps the interconnection of the influence of salt with the degree of hydration in the system. Our findings provide insight into the existing literature data, bear fundamental significance in understanding of hydrated polyelectrolyte assemblies, and suggest a direct means to tailor the mechanical characteristics of PE assemblies via interplay of water and salt.


No capacity limit in attentional tracking: evidence for probabilistic inference under a resource constraint.
Human ability to simultaneously track multiple items declines with set size. This effect is commonly attributed to a fixed limit on the number of items that can be attended to, a notion that is formalized in limited-capacity and slot models. Instead, we propose that observers are constrained by stimulus uncertainty that increases with the number of items but use Bayesian inference to achieve optimal performance. We model five data sets from published deviation discrimination experiments that varied set size, number of deviations, and magnitude of deviation. A constrained Bayesian observer better explains each data set than do the traditional limited-capacity model, the recently proposed slots-plus-averaging model, a fixed-uncertainty Bayesian model, a Bayesian model with capacity limit, and a simple averaging model. This indicates that the notion of limited capacity in attentional tracking needs to be revised. Moreover, it supports the idea that Bayesian optimality of human perception extends to high-level perceptual computations.


Contact pressure and glenohumeral translation following subacromial decompression: how much is enough?
Subacromial decompression is a common surgical procedure that has historically included coracoacromial ligament resection. However, recent reports have advocated preserving the coracoacromial ligament to avoid the potential complication of anterosuperior escape. The optimal subacromial decompression would achieve a smooth coracoacromial arch and decreased rotator cuff contact pressures while preserving the function of the arch in glenohumeral stability. We hypothesized that a subacromial decompression with a limited acromioplasty with preservation of the coracoacromial ligament can decrease extrinsic pressure on the rotator cuff similar to a coracoacromial ligament resection, without altering glenohumeral translation. Three different subacromial decompressions, including a "smooth and move," a limited acromioplasty with coracoacromial ligament preservation, and a coracoacromial ligament resection, were performed on 6 cadaveric specimens with intact rotator cuffs. Glenohumeral translation and peak rotator cuff pressure during abduction were recorded. No change in translation was observed after a smooth and move or a limited acromioplasty. Compared to baseline specimens, anterosuperior translation was increased at 30° of abduction following coracoacromial ligament resection (P<.05). Baseline rotator cuff pressure was greatest during abduction with the arm in 30° of internal rotation. Peak rotator cuff pressure decreased up to 32% following a smooth and move, up to 64% following a limited acromioplasty, and up to 72% following a coracoacromial ligament resection. Based on the present study, a limited acromioplasty with coracoacromial ligament preservation may best provide decompression of the rotator cuff while avoiding potential anterosuperior glenohumeral translation.


Using phase relations to identify potential mechanisms for metabolic oscillations in isolated β-cell mitochondria.
There is a great deal of evidence for the existence of metabolic oscillations in pancreatic β-cells. Mechanisms that have been proposed for these oscillations include glycolytic oscillations; oscillations due to the feedback of Ca2+ onto the mitochondrial inner membrane and on dehydrogenases; and oscillations intrinsic to the tricarboxylic (TCA) cycle or the downstream reactions of oxidative phosphorylation. MacDonald and co-workers (J. Biol. Chem., 278:51894-51900, 2003) showed examples of oscillations in TCA intermediates in isolated mitochondria from liver cells and pancreatic β-cells. These oscillations were clearly not due to oscillations in glycolysis or Ca2+ feedback. In this article we consider several potential mechanisms for these TCA oscillations, using mathematical modeling to determine the phase relations that would result between the citrate and NAD+ concentrations in each case. We demonstrate that negative feedback at only one feedback point, isocitrate dehydrogenase, produces the correct phase relation if oscillations are intrinsic to the TCA cycle. Alternatively, the correct phase relation results if oscillations are due to oscillations in oxidative phosphorylation feeding back onto the TCA cycle. This analysis shows that the observed phase relation between citrate and NAD(P) places strict limits on the potential mechanism for the metabolic oscillations in isolated mitochondria that were observed by MacDonald and co-workers.


Extracorporeal membrane oxygenation for severe Middle East respiratory syndrome coronavirus.
Middle East respiratory syndrome (MERS) is caused by a coronavirus (MERS-CoV) and is characterized by hypoxemic respiratory failure. The objective of this study is to compare the outcomes of MERS-CoV patients before and after the availability of extracorporeal membrane oxygenation (ECMO) as a rescue therapy in severely hypoxemic patients who failed conventional strategies. We collected data retrospectively on MERS-CoV patients with refractory respiratory failure from April 2014 to December 2015 in 5 intensive care units (ICUs) in Saudi Arabia. Patients were classified into two groups: ECMO versus conventional therapy. Our primary outcome was in-hospital mortality; secondary outcomes included ICU and hospital length of stay. Thirty-five patients were included; 17 received ECMO and 18 received conventional therapy. Both groups had similar baseline characteristics. The ECMO group had lower in-hospital mortality (65 vs. 100%, P = 0.02), longer ICU stay (median 25 vs. 8 days, respectively, P < 0.01), and similar hospital stay (median 41 vs. 31 days, P = 0.421). In addition, patients in the ECMO group had better PaO2/FiO2 at days 7 and 14 of admission to the ICU (124 vs. 63, and 138 vs. 36, P < 0.05), and less use of norepinephrine at days 1 and 14 (29 vs. 80%; and 36 vs. 93%, P < 0.05). ECMO use, as a rescue therapy, was associated with lower mortality in MERS patients with refractory hypoxemia. The results of this, largest to date, support the use of ECMO as a rescue therapy in patients with severe MERS-CoV.


Electronic Knowledge Resources and Point-of-Care Learning: A Scoping Review.
The authors sought to summarize quantitative and qualitative research addressing electronic knowledge resources and point-of-care learning in a scoping review. The authors searched MEDLINE, Embase, PsycINFO, and the Cochrane Database for studies addressing electronic knowledge resources and point-of-care learning. They iteratively revised inclusion criteria and operational definitions of study features and research themes of interest. Two reviewers independently performed each phase of study selection and data extraction. Of 10,811 studies identified, 305 were included and reviewed. Most studies (225; 74%) included physicians or medical students. The most frequently mentioned electronic resources were UpToDate (88; 29%), Micromedex (59; 19%), Epocrates (50; 16%), WebMD (46; 15%), MD Consult (32; 10%), and LexiComp (31; 10%). Eight studies (3%) evaluated electronic resources or point-of-care learning using outcomes of patient effects, and 36 studies (12%) reported objectively measured clinician behaviors. Twenty-five studies (8%) examined the clinical or educational impact of electronic knowledge resource use on patient care or clinician knowledge, 124 (41%) compared use rates of various knowledge resources, 69 (23%) examined the quality of knowledge resource content, and 115 (38%) explored the process of point-of-care learning. Two conceptual clarifications were identified, distinguishing the impact on clinical or educational outcomes versus the impact on test setting decision support, and the quality of information content versus the correctness of information obtained by a clinician-user. Research on electronic knowledge resources is dominated by studies involving physicians and evaluating use rates. Studies involving nonphysician users, and evaluating resource impact and implementation, are needed.


Expression of the atrial-specific myosin heavy chain AMHC1 and the establishment of anteroposterior polarity in the developing chicken heart.
A unique myosin heavy chain cDNA (AMHC1), which is expressed exclusively in the atria of the developing chicken heart, was isolated and used to study the generation of diversified cardiac myocyte cell lineages. The pattern of AMHC1 gene expression during heart formation was determined by whole-mount in situ hybridization. AMHC1 is first activated in the posterior segment of the heart when these myocytes initially differentiate (Hamburger and Hamilton stage 9+). The anterior segment of the heart at this stage does not express AMHC1 although the ventricular myosin heavy chain isoform is strongly expressed beginning at stage 8+. Throughout chicken development, AMHC1 continues to be expressed in the posterior heart tube as it develops into the diversified atria. The early activation of AMHC1 expression in the posterior cardiac myocytes suggests that the heart cells are diversified when they differentiate initially and that the anterior heart progenitors differ from the posterior heart progenitors in their myosin isoform gene expression. The expression domain of AMHC1 can be expanded anteriorly within the heart tube by treating embryos with retinoic acid as the heart primordia fuse. Embryos treated with retinoic acid prior to the initiation of fusion of the heart primordia express AMHC1 throughout the entire heart-forming region and fusion of the heart primordia is inhibited. These data indicate that retinoic acid treatment produces an expansion of the posterior (atrial) domain of the heart and suggests that diversified fates of cardiomyogenic progenitors can be altered.


Can Diet Help Non-Obese Individuals with Non-Alcoholic Fatty Liver Disease (NAFLD)?
Subjects diagnosed with non-alcoholic fatty liver disease (NAFLD) or hepatic steatosis are usually obese or overweight. NAFLD has also been reported in many non-obese healthy subjects as an incidental finding during imaging. Subjects with early-stage NAFLD who are otherwise healthy are often left unmanaged in current clinical practice; it is not clear if an early intervention in those individuals would be of any benefit in preventing NAFLD progression to more serious conditions. Since many of these subjects are non-alcoholic and have a normal body mass index (BMI), an intensive lifestyle change program is not usually recommended. This report presents an otherwise healthy non-alcoholic subject with incidental NAFLD having a normal BMI and a waist circumference below 90 cm who successfully reversed his condition by undertaking a lifestyle intervention. The case report is expected to encourage large cohort studies to substantiate the benefits of dietary interventions in alleviating hepatic steatosis among non-obese individuals.


Impact of acute stroke unit on hospital length of stay.
Organized inpatient stroke care (stroke unit: SU) has become the gold standard in stroke care nowadays. The main purpose of this study was to evaluate the effects of implementing an acute stroke unit on the hospital length-of-stay (LoS) on the basis of discharge planning services. Oxford Radcliffe Hospital Trust (ORHT) founded an acute SU in 2005 and closed the rehabilitative stroke wards which have operated for years. Data of acute adult stroke patients admitted to ORHT from 2004 to September of 2005 were sorted to explore the impact of the acute SU on hospital LoS in addition to an active discharge planning program. In total, admission records of 1296 acute stroke patients were identified with the mean age of 73.9+/-13.3 years (range: 20-96 years) and the mean LoS of 25.6+/-37.1 days (median: 11 days). Regarding to the discharge destinations, 25.4% of them died, 48.8% patients returned home, 21.4% patients to community hospitals and 4.3% to various forms of institutional care. The SU admission rate among all acute stroke patients was 37.6%. The hospital LoS of acute stroke patients in ORHT was significantly shortened after the acute stroke unit was started in January 2005 (23.9+/-34.9 days in 2004 vs. 19.3+/-23.0 days in 2005, p=0.038) whilst the stroke mortality rate remained similar. Moreover, in-hospital rehabilitation rate (in geriatric wards of ORHT or community hospitals) was significantly reduced (30.4% vs. 40.9%, p<0.001). The acute SU showed an additional reduction of hospital LoS to the discharge planning program and significantly reduced the needs of in-hospital rehabilitation. Further investigation is needed to explore the impact of acute SU on patients' functional status.


Pigmentation of White, Brown, and Green Chicken Eggshells Analyzed by Reflectance, Transmittance, and Fluorescence Spectroscopy.
We report on the reflectance, transmittance and fluorescence spectra (λ=200-1200 nm) of four types of chicken eggshells (white, brown, light green, dark green) measured in situ without pretreatment and after ablation of 20-100 μm of the outer shell regions. The color pigment protoporphyrin IX (PPIX) is embedded in the protein phase of all four shell types as highly fluorescent monomers, in the white and light green shells additionally as non-fluorescent dimers, and in the brown and dark green shells mainly as non-fluorescent poly-aggregates. The green shell colors are formed from an approximately equimolar mixture of PPIX and biliverdin. The axial distribution of protein and colorpigments were evaluated from the combined reflectances of both the outer and inner shell surfaces, as well as from the transmittances. For the data generation we used the radiative transfer model in the random walk and Kubelka-Munk approaches.


Medullary Endocannabinoids Contribute to the Differential Resting Baroreflex Sensitivity in Rats with Altered Brain Renin-Angiotensin System Expression.
CB1 cannabinoid receptors are expressed on vagal afferent fibers and neurons within the solitary tract nucleus (NTS), providing anatomical evidence for their role in arterial baroreflex modulation. To better understand the relationship between the brain renin-angiotensin system (RAS) and endocannabinoid expression within the NTS, we measured dorsal medullary endocannabinoid tissue content and the effects of CB1 receptor blockade at this brain site on cardiac baroreflex sensitivity (BRS) in ASrAOGEN rats with low glial angiotensinogen, normal Sprague-Dawley rats and (mRen2)27 rats with upregulated brain RAS expression. Mass spectrometry revealed higher levels of the endocannabinoid 2-arachidonoylglycerol in (mRen2)27 compared to ASrAOGEN rats (2.70 ± 0.28 vs. 1.17 ± 0.09 ng/mg tissue; P < 0.01), while Sprague-Dawley rats had intermediate content (1.85 ± 0.27 ng/mg tissue). Microinjection of the CB1receptor antagonist SR141716A (36 pmol) into the NTS did not change cardiac BRS in anesthetized Sprague-Dawley rats (1.04 ± 0.05 ms/mmHg baseline vs. 1.17 ± 0.11 ms/mmHg after 10 min). However, SR141716A in (mRen2)27 rats dose-dependently improved BRS in this strain: 0.36 pmol of SR141716A increased BRS from 0.43 ± 0.03 to 0.71 ± 0.04 ms/mmHg (P < 0.001), and 36 pmol of SR141716A increased BRS from 0.47 ± 0.02 to 0.94 ± 0.10 ms/mmHg (P < 0.01). In contrast, 0.36 pmol (1.50 ± 0.12 vs. 0.86 ± 0.08 ms/mmHg; P < 0.05) and 36 pmol (1.38 ± 0.16 vs. 0.46 ± 0.003 ms/mmHg; P < 0.01) of SR141716A significantly reduced BRS in ASrAOGEN rats. These observations reveal differential dose-related effects of the brain endocannabinoid system that influence cardiovagal BRS in animals with genetic alterations in the brain RAS.


Elderly recipients of liver transplantation: impact of age and psychosocial variables on outcome.
With expanding experience and success of liver transplantation, increasing numbers of elderly candidates await and undergo liver transplantation. There is accumulating evidence that graft survival and mortality does not appear to differ significantly between the young and carefully selected elderly liver transplantation recipients. Although existing evidence suggests that psychosocial factors impact outcomes after liver transplantation in general, no such information is available specifically for elderly (age ≥65 years) liver transplantation recipients. We conducted a broad medical literature review of outcome studies of elderly liver transplantation recipients. In this review article, we summarize the findings and comment on psychosocial variables included in these studies. Ten outcome studies have reported on the impact of age on the liver transplantation outcomes. There is increasing evidence of favorable outcomes in elderly liver transplantation recipients. Few of these studies include measures of quality of life, functional improvement and other psychosocial variables. Very limited information is available about the impact of psychosocial factors on outcomes in elderly liver transplantation recipients. This dearth of information represents a critical gap in our knowledge and has implications for optimal candidate selection and outcomes after liver transplantation.


Outcomes After Elevation of Serratus Anterior Fascia During Prosthetic Breast Reconstruction.
Achieving optimal inferolateral coverage is critical to successful prosthetic breast reconstruction. Serratus anterior fascia (SF) elevation, a promising alternative to muscle flaps and acellular dermis (ADM), has not been rigorously studied. This study evaluates complication rates after mastectomy and immediate tissue expander (TE) coverage using SF, relative to other existing methods of reconstruction. Retrospective review of consecutive patients undergoing mastectomy with immediate TE reconstruction over 10 years at 1 institution was performed. Patients with serratus muscle (SM) or SF elevation were analyzed. ADM reconstructions were used for comparative analysis only. Relevant demographic and clinical data were recorded. Complications were categorized by type and end outcome, including nonoperative (no further surgery), operative (surgery except explantation), and explantation. The SM and SF elevation was performed in 375 (487 breasts) and 177 (255 breasts) patients, respectively. Mean follow-up was 43.8 months. The SM and SF patients were demographically similar, but SF had higher intraoperative fill volumes (P < 0.0001) and required fewer postoperative expansions (P < 0.0001). There were no differences in complications between SM and SF patients. Regression analysis, adjusted for several variables, revealed that SF was not an independent risk factor for complications. The ADM- and SF-assisted reconstruction also showed no differences in outcomes. Our review demonstrates that SF elevation is a safe, feasible alternative for achieving inferolateral coverage during prosthetic breast reconstruction. Furthermore, this technique allows for greater fill volumes and less expansions than SM. As a readily available alternative to muscle flaps and ADM, SF elevation should be considered integral to any prosthetic breast reconstruction algorithm.


Spermiogenesis and spermatozoon ultrastructure of Hunterella nodulosa (Cestoda: Caryophyllidea), a monozoic parasite of suckers (Catostomidae) in North America.
Spermiogenesis and ultrastructure of mature spermatozoon of the caryophyllidean cestode Hunterella nodulosa, a parasite of suckers (Catostomidae), have been studied by transmission electron microscopy. This monozoic tapeworm is unique in its mode of attachment and represents the second North American species studied. The process of spermiogenesis of H. nodulosa follows the general pattern already described in other caryophyllideans. The most characteristic feature is the presence of a slight rotation of the flagellar bud, which seems to be a typical character of spermiogenesis in this cestode group. The mature spermatozoon of H. nodulosa is characterized by the presence of one axoneme of 9 + "1" type of the trepaxonematan flatworms surrounded by a semi-arc of cortical microtubules in its anterior extremity, parallel nucleus and cortical microtubules arranged in a parallel pattern, which corresponds to the Type III pattern of cestode spermatozoa according to Levron et al. (2010). Comparison of the present data with those available for other caryophyllideans did not reveal substantial differences, even though they belong to different families, infect different hosts (catostomid, cyprinid and siluriform fishes) and occur in distant zoogeographical regions. This indicates uniformity of the process of sperm formation and spermatozoon ultrastructure in one of the evolutionarily most ancient groups of tapeworms.


Self-images play a causal role in social phobia.
Patients with social phobia often experience negative self-images in social situations. The current study investigated whether negative self-images have a causal role in maintaining social phobia. Patients with social phobia participated twice in a conversation with a stranger, once whilst holding their usual negative self-image in mind and once whilst holding a less negative (control) self-image in mind, with order counterbalanced across participants. Compared to the control image condition, when participants held the negative image in mind they experienced greater anxiety, rated their anxiety symptoms as being more visible, and rated their performance as poorer. An assessor who did not know which image was being held also rated participants' anxiety as more evident and their behaviour as less positive when the negative image was being held in mind. Finally, when participant and assessor ratings were compared, participants underestimated their performance and overestimated the visibility of their anxiety to a significantly greater extent in the negative imagery condition. Taken together, these results support the hypothesis that negative self-imagery has a causal role in maintaining social phobia.


The Female Athlete Body (FAB) study: Rationale, design, and baseline characteristics.
Eating Disorders (EDs) are serious psychiatric illnesses marked by psychiatric comorbidity, medical complications, and functional impairment. Research indicates that female athletes are often at greater risk for developing ED pathology versus non-athlete females. The Female Athlete Body (FAB) study is a three-site, randomized controlled trial (RCT) designed to assess the efficacy of a behavioral ED prevention program for female collegiate athletes when implemented by community providers. This paper describes the design, intervention, and participant baseline characteristics. Future papers will discuss outcomes. Female collegiate athletes (N=481) aged 17-21 were randomized by site, team, and sport type to either FAB or a waitlist control group. FAB consisted of three sessions (1.3h each) of a behavioral ED prevention program. Assessments were conducted at baseline (pre-intervention), post-intervention (3weeks), and six-, 12-, and 18-month follow-ups. This study achieved 96% (N=481) of target recruitment (N=500). Few group differences emerged at baseline. Total sample analyses revealed moderately low baseline instances of ED symptoms and clinical cases. Health risks associated with EDs necessitate interventions for female athletes. The FAB study is the largest existing RCT for female athletes aimed at both reduction of ED risk factors and ED prevention. The methods presented and population recruited for this study represent an ideal intervention for assessing the effects of FAB on both the aforementioned outcomes. We anticipate that findings of this study (reported in future papers) will make a significant contribution to the ED risk factor reduction and prevention literature.


Hydrodilatation (distension arthrography): a long-term clinical outcome series.
To describe and compare the medium to long-term effectiveness of hydrodilatation and post-hydrodilatation physiotherapy in patients with primary and secondary glenohumeral joint contracture associated with rotator cuff pathology. Patients with primary and secondary glenohumeral contractures associated with rotator cuff pathology were recruited into a 2-year study. They all underwent hydrodilatation, followed by a structured physiotherapy programme. Patients were assessed at baseline, 3 days, 1 week, 3 months, 1 year and 2 years after hydrodilatation with primary outcome measures (Shoulder Pain and Disability Index, Shoulder Disability Index and percentage rating of "normal" function; SD%) and secondary outcome measures (range of shoulder abduction, external rotation and hand behind back). Comparisons in recovery were made between the primary and secondary glenohumeral contracture groups at all timeframes and for all outcome measures. A total of 53 patients (23 with primary and 30 with secondary glenohumeral contractures) were recruited into the study. At the 2-year follow-up, 12 patients dropped out from the study. At baseline, the two contracture groups were similar with respect to their demographic and physical characteristics. The two groups of patients recovered in a similar fashion over the 2-year follow-up period. A significant improvement was observed in all outcomes measures over this period (p<0.01), so that both function and range of movement increased. The rate of improvement was dependent on the outcome measure that was used. Hydrodilatation and physiotherapy increase shoulder motion in individuals with primary and secondary glenohumeral joint contracture associated with rotator cuff pathology. This benefit continues to improve or is maintained in the long term, up to 2 years after hydrodilatation.


Effect of Silicon Absorption on Soybean Resistance to Phakopsora pachyrhizi in Different Cultivars.
Silicon (Si) is recognized for its prophylactic role in alleviating diseases when absorbed by plants and has been proposed as a possible solution against soybean rust, caused by Phakopsora pachyrhizi. However, little is known about its potential effects on soybean (Glycine max) because the plant's ability to absorb Si is poorly defined. In this work, our objectives were to evaluate and quantify the absorption of Si in leaves of different soybean cultivars and to determine if such absorption was able to enhance resistance to soybean rust. In a first set of experiments with cv. Williams 82, hydroponic plants were supplied or not with Si and inoculated with urediniospores of P. pachyrhizi. Chemical analyses revealed no significant differences in the plants' Si content regardless of the treatment, which translated into no effect on rust incidence. However, in a second set of experiments with different cultivars, plants of Korean cultivar Hikmok sorip absorbed nearly four times more Si than those of Williams 82. At the same time, plants from this cultivar exhibited a near absence of disease symptoms when supplied with Si. This resistance appeared to be the result of hypersensitive (HR) reactions that were triggered when plants were fed with Si. These results support the concept that a plant's innate ability to absorb Si will dictate the benefits conferred by a treatment with Si and provide evidence that Si can protect soybean plants against soybean rust through mediated resistance.


Motor learning strategies in basketball players and its implications for ACL injury prevention: a randomized controlled trial.
Adding external focus of attention (EF, focus on the movement effect) may optimize current anterior cruciate ligament (ACL) injury prevention programmes. The purpose of the current study was to investigate the effects of an EF, by a visual stimulus and an internal focus, by a verbal stimulus during unexpected sidestep cutting in female and male athletes and how these effects remained over time. Ninety experienced basketball athletes performed sidestep cutting manoeuvres in three sessions (S1, S2 and S3). In this randomized controlled trial, athletes were allocated to three groups: visual (VIS), verbal (VER) and control (CTRL). Kinematics and kinetics were collected at the time of peak knee frontal plane moment. Males in the VIS group showed a larger vertical ground reaction force (S1: 25.4 ± 3.1 N/kg, S2: 25.8 ± 2.9 N/kg, S3: 25.2 ± 3.2 N/kg) and knee flexion moments (S1: -3.8 ± 0.9 Nm/kg, S2: -4.0 ± 1.2 Nm/kg, S3: -3.9 ± 1.3 Nm/kg) compared to the males in the VER and CTRL groups and to the females in the VIS group (p < 0.05). Additionally, the males in the VIS group reduced knee valgus moment and the females in the VER group reduced knee varus moment over time (n.s.). Male subjects clearly benefit from visual feedback. Females may need different feedback modes to learn a correct movement pattern. Sex-specific learning preferences may have to be acknowledged in day by day practice. Adding video instruction or feedback to regular training regimens when teaching athletes safe movement patterns and providing individual feedback might target suboptimal long-term results and optimize ACL injury prevention programmes. I.


Gas permeability and selectivity of cellulose nanocrystals films (layers) deposited by spin coating.
Cellulose nanocrystals (CNC) were extracted from a cellulose residue using two different acid hydrolysis procedures. CNC extracted with sulfuric acid (CNC(S)) showed higher surface charge (339 μmol/g) compared with crystals extracted with hydrochloric acid (CNC(HCl)). Spin-coated films with two different configurations were prepared; the first with alternate layers of poly(allylamine hydrochloride) (PAHCl) and CNC, and the second with a single layer of PAHCl coated with multilayers of CNC. Film characteristics such as roughness, thickness, contact angle, orientation, gas permeability and gas selectivity were studied. Optical microscopy showed more homogeneous films of CNC(S) compared to CNC(HCl). The surface charge of the crystals impacted the films' hydrophobicity, being highest for 25 alternate layers of PAHCl and CNC(HCl). The gas permeability coefficient was different for each film, depending primarily on the surface charge of the crystals and secondly on the film configuration. The films made with CNC(HCl) displayed gas barriers with nitrogen and oxygen, and gas selectivity with some gas combinations. CNC(S) films did not show gas selectivity. These results indicate that CNC with low surface charge can be further developed for gas separation and barrier applications.


Minority Physician Assistant Faculty: A Phenomenological Assessment of Factors Leading to Retention in the Faculty Role.
Improving racial and ethnic diversity in the physician assistant (PA) profession is important to providing better care for underserved communities. The recruitment and retention of minority PA faculty is one aspect of helping to attract and retain a more diverse student body. Previous research has indicated that minority status is associated with the increased attrition of PA faculty but has not provided insight into the specific factors involved in the retention or attrition of minority PA faculty. The purpose of this qualitative research study was to describe the experience of minority PA faculty through a critical race theory lens. We used a phenomenological approach using structured interviews of minority PA faculty. Better understanding of the experience of minority PA faculty might lead to improved efforts at recruiting and supporting a more diverse faculty workforce. We conducted 13 interviews of PA faculty representing a variety of underrepresented minorities, geographic regions, types of schools, and stages of their careers. Major themes that emerged across the participants' experiences included opportunities for success in the form of both internal and external support systems and mentorship. As a corollary, barriers to the retention of minority PA faculty including a lack of institutional support, gaps in mentorship, and lack of a solid support network were cited.


Mortality amongst participants in Vasaloppet: a classical long-distance ski race in Sweden.
The aim of this study was to assess mortality amongst participants in long-distance ski races during the Vasaloppet week. We considered the 90 km races for men and 90 or 30 km for women. The vast majority of the participants in these races are not competing on the elite level. It is assumed, however, that they have to undergo regular physical training during a long period of time in order to successfully finish the race. The cohort study consisted of 49 219 men and 24 403 women, who participated in any of the races during 1989-1998. All subjects were followed up in the National-Cause-of-Death-Register until 31 December 1999. We computed the standardized mortality ratios (SMRs) adjusting for age and calendar year. Overall, 410 deaths occurred, compared with 850.6 expected, yielding an SMR of 0.48 [95% confidence interval (CI) 0.44-0.53]. Low SMRs were found in all age groups in both men and women and in all groups after categorization by finishing time and number of races. The lowest SMRs were found amongst older participants and in those who participated in several races. A decreased mortality was observed in all major diagnostic groups, namely cancers (SMR = 0.61; 95% CI 0.52-0.71), diseases of the circulatory system (SMR = 0.43; 95% CI 0.35-0.51), and injuries and poisoning (SMR = 0.73; 95% CI 0.60-0.89). For lung cancer the SMR was 0.22, but even after exclusion of lung cancer the all-cancer mortality was low (SMR = 0.72; 95% CI 0.59-0.86). We conclude that participants in long-distance skiing races, which demand prolonged regular physical training, have low mortality. The extent to which this is due to physical activity, related lifestyle factors, genetics or selection bias is yet to be assessed.


Enhancement of the solubility and antioxidant capacity of α-linolenic acid using an oil in water microemulsion.
The applications of α-linolenic acid (ALA) in the food industry are restricted due to its poor water solubility and antioxidant stability. This study concentrates on developing an ALA-loaded microemulsion (ALA-ME) to enhance its solubility and antioxidant capacity. The formulation of the microemulsion was investigated based on pseudoternary phase diagrams. The ALA-ME was characterized by using electrical conductivity, viscosity and transmission electron microscopy (TEM). The microstructure of the ALA-ME was probed using nuclear magnetic resonance (1H-NMR). The results proved that ALA-ME consisted of spheroidal droplets with 20-40 nm diameter. A structural transformation from water in oil (W/O) to oil in water (O/W) occurred, as seen from the electrical conductivity determination. The 1H-NMR results revealed a transition of the ALA position encapsulated from the core area of the microemulsion to the lipophilic layer of the surfactant. Furthermore, two microstructural models of ALA-ME were proposed. The antioxidant evaluation demonstrated that the ALA antioxidant capacity in microemulsions was enhanced to about 80% compared with that of ALA in oil solution.


Anti-inflammatory effect of recreational exercise in TNBS-induced colitis in rats: role of NOS/HO/MPO system.
There are opposite views in the available literature: Whether physical exercise has a protective effect or not on the onset of inflammatory bowel disease (IBD). Therefore, we investigated the effects of recreational physical exercise before the induction of colitis. After 6 weeks of voluntary physical activity (running wheel), male Wistar rats were treated with TNBS (10 mg). 72 hrs after trinitrobenzene sulphonic acid (TNBS) challenge we measured colonic gene (TNF-α, IL-1β, CXCL1 and IL-10) and protein (TNF-α) expressions of various inflammatory mediators and enzyme activities of heme oxygenase (HO), nitric oxide synthase (NOS), and myeloperoxidase (MPO) enzymes. Wheel running significantly increased the activities of HO, constitutive NOS (cNOS) isoform. Furthermore, 6 weeks of running significantly decreased TNBS-induced inflammatory markers, including extent of lesions, severity of mucosal damage, and gene expression of IL-1β, CXCL1, and MPO activity, while IL-10 gene expression and cNOS activity were increased. iNOS activity decreased and the activity of HO enzyme increased, but not significantly, compared to the sedentary TNBS-treated group. In conclusion, recreational physical exercise can play an anti-inflammatory role by downregulating the gene expression of proinflammatory mediators, inducing anti-inflammatory mediators, and modulating the activities of HO and NOS enzymes in a rat model of colitis.


Impact of second-line antiretroviral regimens on lipid profiles in an African setting: the DART trial sub-study.
Increasing numbers of HIV-infected patients in sub-Saharan Africa are exposed to antiretroviral therapy (ART), but there are few data on lipid changes on first-line ART, and even fewer on second-line. DART was a randomized trial comparing monitoring strategies in Ugandan/Zimbabwean adults initiating first-line ART and switching to second-line at clinical/immunological failure. We evaluated fasting lipid profiles at second-line initiation and ≥48 weeks subsequently in stored samples from Zimbabwean patients switching before 18 September 2006. Of 91 patients switched to second-line ART, 65(73%) had fasting samples at switch and ≥48 weeks, 14(15%) died or were lost <48 weeks, 10(11%) interrupted ART for >14 days and 2(2%) had no samples available. 56/65(86%) received ZDV/d4T + 3TC + TDF first-line, 6(9%) ZDV/d4T + 3TC + NVP and 3(5%) ZDV + 3TC with TDF and NVP. Initial second-line regimens were LPV/r + NNRTI in 27(41%), LPV/r + NNRTI + ddI in 33(50%) and LPV/r + TDF + ddI/3TC/ZDV in 6(9%). At second-line initiation median (IQR) TC, LDL-C, HDL-C and TG (mmol/L) were 3.3(2.8-4.0), 1.7(1.3-2.2), 0.7(0.6-0.9) and 1.1(0.8-1.9) respectively. Levels were significantly increased 48 weeks later, by mean (SE) +2.0(0.1), +1.1(0.1), +0.5(0.05) and +0.4(0.2) respectively (p < 0.001; TG p = 0.01). 3% at switch vs 25% 48 weeks later had TC >5.2 mmol/L; 3% vs 25% LDL-C >3.4 mmol/L and 91% vs 41% HDL-C <1.1 mmol/L (p < 0.001). Similar proportions had TG >1.8 mmol/L (0 vs 3%) and TC/HDL-C ≥5 (40% vs 33%) (p > 0.15). Modest lipid elevations were observed in African patients on predominantly LPV/r + NNRTI-based second-line regimens. Routine lipid monitoring during second-line ART regimens may not be warranted in this setting but individual cardiovascular risk assessment should guide practice.


MicroRNA signatures associated with thioacetamide-induced liver fibrosis in mice.
Multiple etiologies of liver injury are associated with fibrosis in which the key event is the activation of hepatic stellate cells (HSCs). Although microRNAs (miRNAs) are reportedly involved in fibrogenesis, the complete array of miRNA signatures associated with the disease has yet to be elucidated. Here, deep sequencing analysis revealed that compared to controls, 80 miRNAs were upregulated and 21 miRNAs were downregulated significantly in the thioacetamide (TAA)-induced mouse fibrotic liver. Interestingly, 58 of the upregulated miRNAs were localized to an oncogenic miRNA megacluster upregulated in liver cancer. Differential expression of some of the TAA-responsive miRNAs was confirmed, and their human orthologs were similarly deregulated in TGF-β1-activated HSCs. Moreover, a functional analysis of the experimentally validated high-confidence miRNA targets revealed significant enrichment for the GO terms and KEGG pathways involved in HSC activation and liver fibrogenesis. This is the first comprehensive report of miRNAs profiles during TAA-induced mouse liver fibrosis.


Structure of the proton-gated urea channel from the gastric pathogen Helicobacter pylori.
Half the world's population is chronically infected with Helicobacter pylori, causing gastritis, gastric ulcers and an increased incidence of gastric adenocarcinoma. Its proton-gated inner-membrane urea channel, HpUreI, is essential for survival in the acidic environment of the stomach. The channel is closed at neutral pH and opens at acidic pH to allow the rapid access of urea to cytoplasmic urease. Urease produces NH(3) and CO(2), neutralizing entering protons and thus buffering the periplasm to a pH of roughly 6.1 even in gastric juice at a pH below 2.0. Here we report the structure of HpUreI, revealing six protomers assembled in a hexameric ring surrounding a central bilayer plug of ordered lipids. Each protomer encloses a channel formed by a twisted bundle of six transmembrane helices. The bundle defines a previously unobserved fold comprising a two-helix hairpin motif repeated three times around the central axis of the channel, without the inverted repeat of mammalian-type urea transporters. Both the channel and the protomer interface contain residues conserved in the AmiS/UreI superfamily, suggesting the preservation of channel architecture and oligomeric state in this superfamily. Predominantly aromatic or aliphatic side chains line the entire channel and define two consecutive constriction sites in the middle of the channel. Mutation of Trp 153 in the cytoplasmic constriction site to Ala or Phe decreases the selectivity for urea in comparison with thiourea, suggesting that solute interaction with Trp 153 contributes specificity. The previously unobserved hexameric channel structure described here provides a new model for the permeation of urea and other small amide solutes in prokaryotes and archaea.


Effects of extended-release niacin with laropiprant in high-risk patients.
Patients with evidence of vascular disease are at increased risk for subsequent vascular events despite effective use of statins to lower the low-density lipoprotein (LDL) cholesterol level. Niacin lowers the LDL cholesterol level and raises the high-density lipoprotein (HDL) cholesterol level, but its clinical efficacy and safety are uncertain. After a prerandomization run-in phase to standardize the background statin-based LDL cholesterol-lowering therapy and to establish participants' ability to take extended-release niacin without clinically significant adverse effects, we randomly assigned 25,673 adults with vascular disease to receive 2 g of extended-release niacin and 40 mg of laropiprant or a matching placebo daily. The primary outcome was the first major vascular event (nonfatal myocardial infarction, death from coronary causes, stroke, or arterial revascularization). During a median follow-up period of 3.9 years, participants who were assigned to extended-release niacin-laropiprant had an LDL cholesterol level that was an average of 10 mg per deciliter (0.25 mmol per liter as measured in the central laboratory) lower and an HDL cholesterol level that was an average of 6 mg per deciliter (0.16 mmol per liter) higher than the levels in those assigned to placebo. Assignment to niacin-laropiprant, as compared with assignment to placebo, had no significant effect on the incidence of major vascular events (13.2% and 13.7% of participants with an event, respectively; rate ratio, 0.96; 95% confidence interval [CI], 0.90 to 1.03; P=0.29). Niacin-laropiprant was associated with an increased incidence of disturbances in diabetes control that were considered to be serious (absolute excess as compared with placebo, 3.7 percentage points; P<0.001) and with an increased incidence of diabetes diagnoses (absolute excess, 1.3 percentage points; P<0.001), as well as increases in serious adverse events associated with the gastrointestinal system (absolute excess, 1.0 percentage point; P<0.001), musculoskeletal system (absolute excess, 0.7 percentage points; P<0.001), skin (absolute excess, 0.3 percentage points; P=0.003), and unexpectedly, infection (absolute excess, 1.4 percentage points; P<0.001) and bleeding (absolute excess, 0.7 percentage points; P<0.001). Among participants with atherosclerotic vascular disease, the addition of extended-release niacin-laropiprant to statin-based LDL cholesterol-lowering therapy did not significantly reduce the risk of major vascular events but did increase the risk of serious adverse events. (Funded by Merck and others; HPS2-THRIVE ClinicalTrials.gov number, NCT00461630.).


Evaluation of trends in marijuana toxicosis in dogs living in a state with legalized medical marijuana: 125 dogs (2005-2010).
To report a correlation between the increased number of medical marijuana licenses and marijuana toxicosis in dogs in a state with legalized marijuana for medical use. Retrospective case series from January 1, 2005 to October 1, 2010. Private specialty referral hospital and a university teaching hospital. A total of 125 client-owned dogs presenting for known or suspected marijuana toxicosis with or without a urine drug screening test (UDST). None. During the study period, 125 dogs were evaluated including 76 dogs with known marijuana exposure or a positive UDST (group 1), 6 dogs with known marijuana ingestion and a negative UDST (group 2), and 43 dogs with known marijuana ingestion that were not tested (group 3). The incidence of marijuana toxicosis presenting to both hospitals increased 4-fold, while the number of people registered for medical marijuana in the state increased 146-fold in the last 5 years. A significant positive correlation was detected between the increase in known/suspected marijuana toxicosis in dogs (groups 1-3) and the increased number of medical marijuana licenses (correlation R coefficient = 0.959, P = 0.002). Two dogs that ingested butter made with medical grade marijuana in baked products died. A significant correlation was found between the number of medical marijuana licenses and marijuana toxicosis cases seen in 2 veterinary hospitals in Colorado. Ingestion of baked goods made with medical grade tetrahydrocannabinol butter resulted in 2 deaths. UDST may be unreliable for the detection of marijuana toxicosis in dogs.


Long-term use of medications and destructive periodontal disease.
Knowledge of typical medication use among patients with chronic periodontitis or destructive periodontal disease is limited. The aim of this study was to associate periodontitis severity with the use of different classes of medications. Patients (N=12,631) who had medical, dental, and pharmaceutical coverage with a health maintenance organization and whose severity of destructive periodontal disease was diagnosed by a dentist or specialist were included in the study. The rate of drug use over 7 years was related to the severity of destructive periodontal disease by means of Poisson regression models. Individuals with moderate to advanced periodontitis had significantly lower fill rates for the respiratory agents (antihistamines: -23%, 95% confidence interval [CI]: -10% to -34%; decongestants: -24%, 95% CI: -13% to -34%; and cough/cold medications: -12%, 95% CI: -3% to -21%) and anti-infective agents (antibiotics: -12%, 95% confidence interval: -6% to -18%; urinary anti-infectives: -36%, 95% CI: -6% to -56%; and topical antibiotics: -18%, 95% CI: -5% to -29%). More severe periodontitis was a marker for reduced medication use for allergies and infections. The associations between destructive periodontal disease, infections, allergies, and the hygiene hypothesis need further exploration.


The role of endonasal surgery in the management of frontoethmoidal osteomas.
Recently endonasal surgery has been considered to be a valuable contribution in the management of paranasal sinus osteoma. A retrospective evaluation study of 34 frontoethmoidal osteomas (23 frontal and 11 ethmoidal osteomas) treated at a tertiary care facility from 1990 to 1999 is presented. Twenty three osteomas (68%) were resected endonasally. Eleven osteomas (32%) were removed using an osteoplastic frontal sinus approach with coronal incision. In 5 cases of huge osteomas originating at the anterior frontal sinus wall, reconstruction of the resected anterior-frontal sinus wall was achieved by autologous outer table grafts harvested from the parietal region. Endoscopic and radiological follow-up ranging from 1 to 32 months showed three incomplete endonasal osteoma resections. Complete osteoma removal was achieved via endonasal revision surgery in two of these cases, while the third small residual osteoma remains under observation. There was no case of osteoplastic osteoma removal where incomplete osteoma resection became obvious during follow-up. Ethmoidal osteomas without extrasinusal extension can be resected endonasally. The endonasal approach should be considered also for frontal sinus osteomas if (1) sufficient frontal sinus access can be achieved endonasally, (2) the osteoma is placed medially to a virtual sagittal plane through the lamina papyracea, and (3) the tumour base is at the inferior part of the posterior frontal sinus wall. We favour the osteoplastic frontal sinus approach with coronal incision if an external approach is required to achieve tumour resection with the best aesthetic results.


Sensitivity of proneural genes to lateral inhibition affects the pattern of primary neurons in Xenopus embryos.
We have compared the roles of XASH-3 and NeuroD, two basic helix-loop-helix transcription factors, in the formation of primary neurons in early Xenopus embryos. When ectopically expressed in Xenopus embryos, XASH-3 and NeuroD induce ectopic primary neurons in very different spatial patterns. We show that the pattern of primary neurons induced by XASH-3 and NeuroD can be accounted for by a difference in their sensitivity to inhibitory interactions mediated by the neurogenic genes, X-Delta-1 and X-Notch-1. Both NeuroD and XASH-3 promote the expression of the inhibitory ligand, X-Delta-1. However, XASH-3 appears to be sensitive to the inhibitory effects of X-Delta-1 while NeuroD is much less so. Consequently only a subset of cells that ectopically express XASH-3 eventually form neurons, giving a scattered pattern, while the ectopic expression of NeuroD leads to a relatively dense pattern of ectopic neurons. We propose that differences in the sensitivity of XASH-3 and NeuroD to lateral inhibition play an important role during their respective roles in neuronal determination and differentiation.


Successful use of extracorporeal membrane oxygenation to treat severe respiratory failure in a pediatric patient with a scald injury.
If ECMO is to be used effectively in pediatric patients, specifically in those with burns, the candidates must be chosen with care. Unlike the situation in neonates, when ECMO is being considered for use in a pediatric patient, no clear set of inclusion or exclusion criteria exists. Evaluation of a pediatric patient for ECMO support is largely based on an assessment of the patient's condition and a center's previous experience with pediatric ECMO. The data that are available through ELSO indicate that survival decreases as the number of days a patient receives mechanical ventilation before the initiation of ECMO increases. The effect of burns on patients' outcomes is unknown. Age, duration of mechanical ventilation, and excision with allografting or homografting of the burns should all be considered before the patient is offered ECMO support. The remaining prognostic signs--duration of ECMO support, frequency of complications, and blood product requirements--are available only after the ECMO course is under way or completed. The success of our center and others in using ECMO to treat respiratory failure associated with burns shows that some patients with burns may benefit from ECMO. Unfortunately, no specific set of criteria exists that would enable ECMO centers to differentiate good candidates from poor ones and thus be able to offer ECMO support with confidence in its benefit for the patient.


Construction of normalized RNA-seq libraries for next-generation sequencing using the crab duplex-specific nuclease.
RNA-seq is a method for studying the transcriptome of cells or tissues by massively parallel sequencing of tens of millions of short DNA fragments. However, the broad dynamic range of gene expression levels, which span more than five orders of magnitude, necessitates considerable over-sequencing to characterize low-abundance RNAs at sufficient depth. Here, we describe a method that enables efficient sequencing of low-abundance RNAs by normalizing or reducing the range spanned by the most abundant RNA species to the least abundant RNA species. This normalization is achieved using an approach that was developed for generating expressed sequence tag (EST) libraries that uses the crab duplex-specific nuclease and exploits the kinetics of DNA annealing. That is, double-stranded cDNA is denatured, then allowed to partially re-anneal, and the most abundant species, which re-anneal most rapidly, are digested with crab duplex-specific nuclease. This procedure substantially decreases the proportion of sequence reads from highly expressed RNAs, facilitating assessment of the full spectrum of the sequence and structure of transcriptomes.


On different facets of regularization theory.
This review provides a comprehensive understanding of regularization theory from different perspectives, emphasizing smoothness and simplicity principles. Using the tools of operator theory and Fourier analysis, it is shown that the solution of the classical Tikhonov regularization problem can be derived from the regularized functional defined by a linear differential (integral) operator in the spatial (Fourier) domain. State-of-the-art research relevant to the regularization theory is reviewed, covering Occam's razor, minimum length description, Bayesian theory, pruning algorithms, informational (entropy) theory, statistical learning theory, and equivalent regularization. The universal principle of regularization in terms of Kolmogorov complexity is discussed. Finally, some prospective studies on regularization theory and beyond are suggested.


Foreign compound metabolism studies with human liver obtained as surgical waste. Relation to donor characteristics and effects of tissue storage.
An investigation has been conducted on the foreign compound-metabolizing activity of human liver collected fresh from surgery or at autopsy from cadavers 3 to 18 hr old, and the effects of low temperature storage on the foreign compound-metabolizing activity of fresh human liver. The enzyme activities studied were microsomal cytochrome P-450 content, biphenyl 4-hydroxylation, benzo-(a)pyrene metabolism, halothane reduction, and 4-hydroxybiphenyl UDP-glucuronosyl transferase, as well as cytosolic thiopurine methyltransferase, thermostable (TS) phenolsulfotransferase, thermolabile (TL) phenolsulfotransferase, and 5-fluorouracil dehydrogenase. Cadaver liver was a poor source of material for metabolism studies with the majority of the enzymes investigated. There was an 84% decrease in the yield of microsomal protein, a 64% decrease in cytochrome P-450 content per mg of microsomal protein, and a 36% decrease in the biphenyl 4-hydroxylase specific activity in human cadaver liver that was a few hours old. UDP-glucuronosyl transferase showed a 70% decrease, TS phenolsulfotransferase a 84% decrease, TL phenolsulfotransferase a 97% decrease, and thiopurine methyltransferase no significant change in specific activity. The loss of activity for many of these enzymes could be simulated by keeping fresh human liver at the temperature of the body after death. Surgical waste provided a good source of fresh, histologically normal, human liver for metabolism studies. Microsomal cytochrome P-450 content showed a significant increase with the age of the liver donor. Metabolism of benzo(a)pyrene to 3-hydroxybenzo(a)pyrene and benzo(a)pyrene-7,8- and -9,10-diols showed up to 136% higher rates in fresh liver microsomes from female donors.(ABSTRACT TRUNCATED AT 250 WORDS)


Z/AP, a double reporter for cre-mediated recombination.
The Cre/loxP site-specific recombination system combined with embryonic stem cell-mediated technologies has greatly expanded our capability to address normal and disease development in mammals using genetic approaches. The success of this emerging technology hinges on the production of Cre-expressing transgenic lines that provide cell type-, tissue-, or developmental stage-specific recombination between loxP sites placed in the genome. Here we describe and characterize the production of a double-reporter mouse line that provides a convenient and reliable readout of Cre recombinase activity. Throughout all embryonic and adult stages, the transgenic animal expresses the lacZ reporter gene before Cre-mediated excision occurs. Cre excision, however, removes the lacZ gene, allowing expression of the second reporter, the human alkaline phosphatase gene. This double-reporter transgenic line is able to indicate the occurrence of Cre excision in an extremely widespread manner from early embryonic to adult lineages. It will be a valuable reagent for the increasing number of investigators taking advantage of the powerful tools provided by the Cre/loxP site-specific recombinase system.


Factor structure of early smoking experiences and associations with smoking behavior: valence or sensitivity model?
The Early Smoking Experience (ESE) questionnaire is the most widely used questionnaire to assess initial subjective experiences of cigarette smoking. However, its factor structure is not clearly defined and can be perceived from two main standpoints: valence, or positive and negative experiences, and sensitivity to nicotine. This article explores the ESE's factor structure and determines which standpoint was more relevant. It compares two groups of young Swiss men (German- and French-speaking). We examined baseline data on 3,368 tobacco users from a representative sample in the ongoing Cohort Study on Substance Use Risk Factors (C-SURF). ESE, continued tobacco use, weekly smoking and nicotine dependence were assessed. Exploratory structural equation modeling (ESEM) and structural equation modeling (SEM) were performed. ESEM clearly distinguished positive experiences from negative experiences, but negative experiences were divided in experiences related to dizziness and experiences related to irritations. SEM underlined the reinforcing effects of positive experiences, but also of experiences related to dizziness on nicotine dependence and weekly smoking. The best ESE structure for predictive accuracy of experiences on smoking behavior was a compromise between the valence and sensitivity standpoints, which showed clinical relevance.


Harmonium: A pulse preserving source of monochromatic extreme ultraviolet (30-110 eV) radiation for ultrafast photoelectron spectroscopy of liquids.
A tuneable repetition rate extreme ultraviolet source (Harmonium) for time resolved photoelectron spectroscopy of liquids is presented. High harmonic generation produces 30-110 eV photons, with fluxes ranging from ∼2 × 10(11) photons/s at 36 eV to ∼2 × 10(8) photons/s at 100 eV. Four different gratings in a time-preserving grating monochromator provide either high energy resolution (0.2 eV) or high temporal resolution (40 fs) between 30 and 110 eV. Laser assisted photoemission was used to measure the temporal response of the system. Vibrational progressions in gas phase water were measured demonstrating the ∼0.2 eV energy resolution.


Origins of enantioselectivity in asymmetric ketone hydrogenation catalyzed by a RuH2(binap)(cydn) complex: insights from a computational study.
In this paper, the origins of enantioselectivity in asymmetric ketone hydrogenation catalyzed by RuH(2)(binap)(cydn) (cydn = trans-1,2-diaminocyclohexane) were discussed. Fifteen substrates involving aromatic, heteroaromatic, olefinic and dialkyl prochiral ketones were used to probe the catalytic mechanism and find an effective way to predict the chirality of the products. The calculated results demonstrate that the hydrogen transfer (HT) step from the Ru complex to the ketone substrate is the chirality-determining step in the H(2)-hydrogenation of ketones. The hydrogenation of aromatic-alkyl ketones can give higher enantiomeric excess (ee) values than that of dialkyl ketones. An interesting intermediate (denoted as ) could be formed if there is an α-hydrogen for R/R' groups of the ketone due to the H(2)-H(α) interaction. Two substituent groups of the ketone could rotate around the C=O axis in two directions, clockwise or counter-clockwise. This rotation, with the big or conjugative substituent group away from/toward the closer binap ligand of the Ru catalyst, will form favorable/unfavorable chiral products with an Re-/Si- intermediate structure. On the contrary, if there is no such α-hydrogen in any substituent group of the ketone, ABS and another intermediate (denoted as INT) would not exist. This study indicates that the conjugative effect of the substituent groups of the ketone play an important role in differentiating the R/R' groups of the ketone, while steric and electrostatic effects contribute to a minor extent. Furthermore, the disparity of the R and R' groups of the ketone is of importance in the enantioselectivity and the favorable chiral alcohol is formed when the structure of the conjugative/big substituent group is away from the closer binap ligand of the RuH(2)(binap)(cydn) catalyst. According to the three factors of the substituent group and the fourth quadrant theory, the enantioselectivity of 91 prochiral ketones catalyzed by a series of Ru catalysts were predicted. All of the predictions are consistent with the experimental results.


Bioactive Silk Coatings Reduce the Adhesion of Staphylococcus aureus while Supporting Growth of Osteoblast-like Cells.
Orthopedic and dental implants are associated with a substantial risk of failure due to biomaterial-associated infections and poor osseointegration. To prevent such outcomes, a coating can be applied on the implant to ideally both reduce the risk of bacterial adhesion and support establishment of osteoblasts. We present a strategy to construct dual-functional silk coatings with such properties. Silk coatings were made from a recombinant partial spider silk protein either alone (silkwt) or fused with a cell-binding motif derived from fibronectin (FN-silk). The biofilm-dispersal enzyme Dispersin B (DspB) and two peptidoglycan degrading endolysins, PlySs2 and SAL-1, were produced recombinantly. A sortase recognition tag (SrtTag) was included to allow site-specific conjugation of each enzyme onto silkwt and FN-silk coatings using an engineered variant of the transpeptidase Sortase A (SrtA*). To evaluate bacterial adhesion on the samples, Staphylococcus aureus was incubated on the coatings and subsequently subjected to live/dead staining. Fluorescence microscopy revealed a reduced number of bacteria on all silk coatings containing enzymes. Moreover, the bacteria were mobile to a higher degree, indicating a negative influence on the bacterial adhesion. The capability to support mammalian cell interactions was assessed by cultivation of the osteosarcoma cell line U-2 OS on dual-functional surfaces, prepared by conjugating the enzymes onto FN-silk coatings. U-2 OS cells could adhere to silk coatings with enzymes and showed high spreading and viability, demonstrating good cell compatibility.


pgaA and pgaB encode two constitutively expressed endopolygalacturonases of Aspergillus niger.
The nucleotide sequence data for pgaA and pgaB have been deposited with the EMBL, GenBank and DDBJ Databases under accession numbers Y18804 and Y18805 respectively. pgaA and pgaB, two genes encoding endopolygalacturonases (PGs, EC 188.8.131.52) A and B, were isolated from a phage genomic library of Aspergillus niger N400. The 1167 bp protein coding region of the pgaA gene is interrupted by one intron, whereas the 1234 bp coding region of the pgaB gene contains two introns. The corresponding proteins, PGA and PGB, consist of 370 and 362 amino acid residues respectively. Northern-blot analysis revealed that pgaA- and pgaB-specific mRNA accumulate in mycelia grown on sucrose. mRNAs are also present upon transfer to media containing D-galacturonic acid and pectin. Recombinant PGA and PGB were characterized with respect to pH optimum, activity on polygalacturonic acid, and mode of action and kinetics on oligogalacturonates of different chain length (n=3-7). At their pH optimum the specific activities in a standard assay for PGA (pH 4.2) and PGB (pH 5.0) were 16.5 mu+kat.mg(-1) and 8.3 mu+kat.mg(-1) respectively. Product progression analysis, using polygalacturonate as a substrate, revealed a random cleavage pattern for both enzymes and indicated processive behaviour for PGA. This result was confirmed by analysis of the mode of action using oligogalacturonates. Processivity was observed when the degree of polymerization of the substrate exceeded 6. Using pectins of various degrees of methyl esterification, it was shown that PGA and PGB both preferred partially methylated substrates.


[Emergencies in total hip replacement].
Over the last three decades total hip replacement became one of the most common surgical procedures in orthopaedic surgery. According to the number of large joint endoprosthesis, hip replacement is on the first place. Lately, the number of revisional and special tumoral endoprosthesis is increasing, with more severe complications. Dislocation is a leading early complication of total hip arthroplasty. Dislocations after primary total hip arthroplasties occur at an overall incidence of 1-3%, and at 15-20% in revision and tumoral procedures. Closed reduction and eventually immobilization is the method of treatment. If closed reposition in not possible, revision surgery must be performed. Periprosthetic fractures are, as every other fracture, indication for a surgical treatment. Depending on the type of fracture the method of treatment is either open reduction and internal fixation or removal of the primary and implantation of revision endoprosthesis. Deep infection following total joint replacement remains one of the most serious complications, often needing surgical treatment. Treatment consists of incision and debridement. If there is a fever, increased erythrocyte sedimentation and CRP with signs of sepsis, endoprosthesis must be removed. A haematoma appearance after surgical procedure is an emergency which needs a surgical treatment--haematoma evacuation in order to prevent further complication, on the first place infection. Fractured endoprosthesis is one of the most severe complication in the total hip replacement, and need to be surgically treated as soon as possible with endoprosthesis replacement. Aseptic loosening can also be considered as a relative emergency in surgical treatment of total hip replacement. Longer waiting for reoperation can cause losing valuable bone mass needed for revisional endoprosthesis implantation and fixation. Although emergencies in hip replacement are not very common, they must be recognized and eventually surgically treated as soon as possible.


Hyperlipidemia in coronary heart disease. I. Lipid levels in 500 survivors of myocardial infarction.
Plasma cholesterol and triglyceride levels were measured after an overnight fast in 500 consecutively studied 3-mo survivors of myocardial infarction. Virtually all patients under 60 yr of age (95% ascertainment) and a randomly chosen group of older survivors admitted to 13 Seattle hospitals during an 11 mo period were included. A comparison of their lipid values with those of 950 controls demonstrated that 31% had hyperlipidemia. These lipid abnormalities were most commonly found in males under 40 yr of age (60% frequency) and in females under 50 yr of age (60% frequency). Elevation in triglyceride levels with (7.8%) or without (15.6%) an associated elevation in cholesterol levels was three times more common in survivors than a high cholesterol level alone (7.6%). These results raise the possibility that hypertriglyceridemia may be as an important a risk factor for coronary atherosclerosis as hypercholesterolemia. The identification of hyperlipidemic survivors of myocardial infarction provided a unique source of probands for family studies designed to disclose the genetic origin of hyperlipidemia in coronary heart disease.


Endothelium-independent contractions of human cerebral arteries in response to vasopressin.
We studied the effects of vasopressin in isolated segments from branches (500-700 micrograms in external diameter) of human middle cerebral arteries obtained during autopsy of 15 patients who had died 3-8 hours before. Paired segments, one normal and the other de-endothelized by gentle rubbing, were mounted for isometric recording of tension in organ baths. In 11 normal segments, vasopressin produced concentration-dependent contractions with an EC50 of 7.0 X 10(-10) M. Removal of the endothelium from 12 segments did not significantly affect vasopressin-induced contractions. Vasopressin produced further contractions in arterial segments with (n = 4) or without (n = 5) endothelium precontracted with KCl. In segments precontracted with prostaglandin F2 alpha, acetylcholine choline caused relaxation only of those with endothelium. At 10(-8) M (n = 11), the vasopressin V-1 receptor antagonist d(CH2)5Tyr(Me)AVP produced a 60-fold shift to the right of the control response curve for vasopressin. Increasing the concentration of the receptor antagonist to 10(-6) M (n = 7) further displaced the control curve in a parallel manner. These results indicate that vasopressin exerts a powerful constrictor action on isolated human cerebral arteries by direct stimulation of V-1 receptors located predominantly on smooth muscle cells. It appears that this contractile response is not modulated by the presence of an intact endothelial cell layer.


Analysis and improvement of motion encoding in magnetic resonance elastography.
Magnetic resonance elastography (MRE) utilizes phase contrast magnetic resonance imaging (MRI), which is phase locked to externally generated mechanical vibrations, to measure the three-dimensional wave displacement field. At least four measurements with linear-independent encoding directions are necessary to correct for spurious phase contributions if effects from imaging gradients are non-negligible. In MRE, three encoding schemes have been used: unbalanced four- and six-point and balanced four-point ('tetrahedral') encoding. The first two sensitize to motion with orthogonal gradients, with the four-point method acquiring a single reference scan without motion sensitization, whereas three additional scans with inverted gradients are used with six-point encoding, leading to two-fold higher displacement-to-noise ratio (DNR) and 50% longer scan duration. Balanced four-point (tetrahedral) encoding encodes along the four diagonals of a cube, with one direction serving as a reference for the other three encoding directions, similar to four-point encoding. The objective of this work is to introduce a theoretical framework to compare different motion sensitization strategies with respect to their motion encoding efficiency in two fundamental encoding limits, the gradient strength limit and the dynamic range limit, which are both placed in relation to conventional gradient recalled echo (GRE)- and spin echo (SE)-based MRE sequences. We apply the framework to the three aforementioned schemes and show that the motion encoding efficiency of unbalanced four- and six-point encoding schemes in the gradient-limited regime can be increased by a factor of 1.5 when using all physical gradient channels concurrently. Furthermore, it is demonstrated that reversing the direction of the reference in balanced four-point (tetrahedral) encoding results in the Hadamard encoding scheme, which leads to increased DNR by 2 compared with balanced four-point encoding and 2.8 compared with unbalanced four-point encoding. As an example, we show that optimal encoding can be utilized to reduce the acquisition time of standard liver MRE in vivo from four to two breath holds.


Screening for Spiritual Struggle in an Adolescent Transgender Clinic: Feasibility and Acceptability.
Spiritual struggles are associated with poorer health outcomes, including depression, which has higher prevalence among transgender individuals than the general population. This study's objective was to improve the quality of care in an outpatient transgender clinic by screening patients and caregivers for spiritual struggle and future intervention. The quality improvement questions addressed were whether screening for spiritual struggle was feasible and acceptable; and whether the sensitivity and specificity of the Rush Protocol were acceptable. Revision of the screening was based on cognitive interviews with the 115 adolescents and caregivers who were screened. Prevalence of spiritual struggle was 38-47%. Compared to the Negative R-COPE, the Rush Protocol screener had sensitivities of 44-80% and specificities of 60-74%. The Rush Protocol was acceptable to adolescents seen in a transgender clinic, caregivers, and clinic staff; was feasible to deliver during outpatient clinic visits, and offers a straightforward means of identifying transgender persons and caregivers experiencing spiritual struggle.


Low CD4/CD8 T-cell ratio associated with inflammatory arthropathy in human T-cell leukemia virus type I Tax transgenic mice.
Human T-cell leukemia virus type I (HTLV-1) can cause an aggressive malignancy known as adult T-cell leukemia/lymphoma (ATL) as well as inflammatory diseases such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). A transgenic mouse that expresses HTLV-1 Tax also develops T-cell leukemia/lymphoma and an inflammatory arthropathy that resembles rheumatoid arthritis. The aim of this study was to identify the primary T-cell subsets involved in the development of arthropathy in Tax transgenic mice. By 24 months of age, Tax transgenic mice developed severe arthropathy with a cumulative incidence of 22.8%. The pathological findings of arthropathy in Tax transgenic mice were similar to those seen in human rheumatoid arthritis or mouse models of rheumatoid arthritis, with synovial proliferation and a positive rheumatoid factor. Before the onset of spontaneous arthropathy, young and old Tax transgenic mice were not sensitive to collagen and did not develop arthritis after immunization with type II collagen. The arthropathic Tax transgenic mice showed a significantly decreased proportion of splenic CD4(+) T cells, whereas the proportion of splenic CD8(+) T cells was increased. Regulatory T cells (CD4(+)CD25(+)Foxp3(+)) were significantly decreased and CD8(+) T cells that expressed the chemokine receptor CCR4 (CD8(+)CCR4(+)) were significantly increased in arthropathic Tax transgenic mice. The expression of tax mRNA was strong in the spleen and joints of arthropathic mice, with a 40-fold increase compared with healthy transgenic mice. Our findings reveal that Tax transgenic mice develop rheumatoid-like arthritis with proliferating synovial cells in the joints; however, the proportion of different splenic T-cell subsets in these mice was completely different from other commonly used animal models of rheumatoid arthritis. The crucial T-cell subsets in arthropathic Tax transgenic mice appear to resemble those in HAM/TSP patients rather than those in rheumatoid arthritis patients.


Whole-body sleeping beauty mutagenesis can cause penetrant leukemia/lymphoma and rare high-grade glioma without associated embryonic lethality.
The Sleeping Beauty (SB) transposon system has been used as a somatic mutagen to identify candidate cancer genes. In previous studies, efficient leukemia/lymphoma formation on an otherwise wild-type genetic background occurred in mice undergoing whole-body mobilization of transposons, but was accompanied by high levels of embryonic lethality. To explore the utility of SB for large-scale cancer gene discovery projects, we have generated mice that carry combinations of different transposon and transposase transgenes. We have identified a transposon/transposase combination that promotes highly penetrant leukemia/lymphoma formation on an otherwise wild-type genetic background, yet does not cause embryonic lethality. Infiltrating gliomas also occurred at lower penetrance in these mice. SB-induced or accelerated tumors do not harbor large numbers of chromosomal amplifications or deletions, indicating that transposon mobilization likely promotes tumor formation by insertional mutagenesis of cancer genes, and not by promoting wide-scale genomic instability. Cloning of transposon insertions from lymphomas/leukemias identified common insertion sites at known and candidate novel cancer genes. These data indicate that a high mutagenesis rate can be achieved using SB without high levels of embryonic lethality or genomic instability. Furthermore, the SB system could be used to identify new genes involved in lymphomagenesis/leukemogenesis.


Vitamin E reduces endosulfan-induced toxic effects on morphology and behavior in early development of zebrafish (Danio rerio).
The aim of this study was to investigate if vitamin E (α-TOC) modulates the developmental toxicity of the pesticide endosulfan (ESF), using a modified zebrafish embryotoxicity test (ZET). Zebrafish (Danio rerio) embryos were exposed from 6 to 72 h post fertilization (hpf) to either ESF (0.1-50 mg/L) or α-TOC (0.01-3 mM) alone or in combination. The effects of these exposures on embryonic morphology, larval behavior and antioxidant gene expression were analyzed. Phenotypic analysis at 48 hpf showed that ESF led to a dose-dependent increase in embryonic deformities, including axis malformations, pericardial edema and reduced pigmentation. Co-exposure of ESF with α-TOC (1-3 mM) significantly (p < 0.05) reduced ESF-induced embryonic malformations. Exposure to solely α-TOC did not affect rates of survival or malformations. Behavior studies showed that ESF caused hyperactivity at 5 days post fertilization, indicating a developmental neurotoxic effect. The ESF-induced hyperactivity was ameliorated by α-TOC. Elevated ESF concentrations caused down-regulation of the antioxidant genes cuzn-sod, gpx1a and cat, suggesting that ESF promoted oxidative stress in the embryos. α-TOC did not prevent the ESF-induced dysregulation of these genes. These results demonstrate that α-TOC protects against phenotypic and behavioral effects caused by ESF but did not rescue ESF-induced aberrations in antioxidant gene expression.


Inhibition of CRM1-mediated nuclear export of transcription factors by leukemogenic NUP98 fusion proteins.
NUP98 is a nucleoporin that plays complex roles in the nucleocytoplasmic trafficking of macromolecules. Rearrangements of the NUP98 gene in human leukemia result in the expression of numerous fusion oncoproteins whose effect on nucleocytoplasmic trafficking is poorly understood. The present study was undertaken to determine the effects of leukemogenic NUP98 fusion proteins on CRM1-mediated nuclear export. NUP98-HOXA9, a prototypic NUP98 fusion, inhibited the nuclear export of two known CRM1 substrates: mutated cytoplasmic nucleophosmin and HIV-1 Rev. In vitro binding assays revealed that NUP98-HOXA9 binds CRM1 through the FG repeat motif in a Ran-GTP-dependent manner similar to but stronger than the interaction between CRM1 and its export substrates. Two NUP98 fusions, NUP98-HOXA9 and NUP98-DDX10, whose fusion partners are structurally and functionally unrelated, interacted with endogenous CRM1 in myeloid cells as shown by co-immunoprecipitation. These leukemogenic NUP98 fusion proteins interacted with CRM1, Ran, and the nucleoporin NUP214 in a manner fundamentally different from that of wild-type NUP98. NUP98-HOXA9 and NUP98-DDX10 formed characteristic aggregates within the nuclei of a myeloid cell line and primary human CD34+ cells and caused aberrant localization of CRM1 to these aggregates. These NUP98 fusions caused nuclear accumulation of two transcription factors, NFAT and NFkappaB, that are regulated by CRM1-mediated export. The nuclear entrapment of NFAT and NFkappaB correlated with enhanced transcription from promoters responsive to these transcription factors. Taken together, the results suggest a new mechanism by which NUP98 fusions dysregulate transcription and cause leukemia, namely, inhibition of CRM1-mediated nuclear export with aberrant nuclear retention of transcriptional regulators.


The effects of coffee on enzymes involved in metabolism of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in rats.
The effects of coffee on the metabolism and genotoxicity of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were investigated. Coffee diminished the bacterial mutagenicity of PhIP in the Ames reversion assay through inhibition of cytochrome P450 1A2 (CYP1A2), a key enzyme involved in the metabolic activation of PhIP. When given as part of the diet (0, 1 or 5% w/w) to male Fischer-344 rats for 2 weeks, coffee affected the expression of hepatic enzymes involved in PhIP metabolism. Coffee increased the expression of CYP1A2 by 16-fold in the 5% coffee-treated group, and approximately half of this inductive effect was attributed to caffeine. Coffee also increased the expression of enzymes involved in the detoxication of PhIP. A 2-fold increase in expression of glutathione S-transferase alpha was observed, UDP-glucuronosyl transferase (UGTs) activities of p-nitrophenol increased 2-fold, while N(2)-and N3-glucuronidation of the genotoxic metabolite 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (HONH-PhIP) increased by 1.3-fold in the 5% coffee-treated over the control group. The amount of PhIP (0.75 mg/kg, 24 h) eliminated in urine as the N(2)-and N3-glucuronide conjugates of HONH-PhIP increased by 1.8- and 2.5-fold, respectively, in the 5% coffee-treated group over control rats, suggesting either increased rates of N-oxidation of PhIP or N-glucuronidation of HONH-PhIP. Despite the strong induction of CYP1A2, there was no increase in PhIP-DNA adduct formation in colon and pancreas while liver adducts decreased by 50% over control animals. These data suggest that the effect of coffee on inhibition of PhIP N-oxidation and ensuing DNA damage is more important in vivo than its effect on induction of PhIP N-hydroxylation.


Nitration of distinct tyrosine residues causes inactivation of histone deacetylase 2.
Histone deacetylases (HDACs) are key molecules involved in epigenetic regulation of gene expression. We have previously demonstrated that oxidative stress caused a reduction in HDAC2, resulting in amplified inflammation and reduced corticosteroid responsiveness. Here we showed nitrative/oxidative stress reduced HDAC2 expression via nitration of distinct tyrosine residues. Peroxynitrite, hydrogen peroxide and cigarette smoke-conditioned medium reduced HDAC2 expression in A549 epithelial cells in vitro. This reduction was due to increased proteasomal degradation following ubiquitination rather than reduction of mRNA expression or stability. HDAC2 was nitrated under nitrative/oxidative stress and in the peripheral lung tissues of smokers and patients with chronic obstructive pulmonary disease. Mutagenesis studies replacing tyrosine (Y) residues with alanine revealed that Y253 is at least partly responsible for the proteasomal degradation of HDAC2 under nitrative stress. Thus, nitration of distinct tyrosine residues modifies both the expression and activity of HDAC2, having an impact on epigenetic regulation.


Identification of three CCp genes in Babesia divergens: novel markers for sexual stages parasites.
Babesia divergens, a tick-borne protozoan parasite of red blood cells, is the main agent of bovine and human babesiosis in Europe. Very few data are available concerning its life cycle and sexual reproduction inside the tick vector, Ixodes ricinus. The aim of this study was to define some markers of the B.divergens sexual stage. An in silico post-genomic approach was used to analyze genomic, transcriptomic and proteomic data and to select specific sexual stage proteins of the related apicomplexan genus Plasmodium. Three proteins, based on sequence identity between the available genomes of Plasmodium and Babesia spp., were chosen, as members of a highly conserved and apicomplexan sexual stages specific protein family (CCp) potentially involved in adhesive functions. Degenerate primers were used to amplify and clone three B.divergens orthologs (bdccp1, bdccp2, and bdccp3) corresponding to newly identified genes in this parasite. The opportunities offered by such markers to study parasite development in its vector are discussed.


The autoimmune-associated genetic variant PTPN22 R620W enhances neutrophil activation and function in patients with rheumatoid arthritis and healthy individuals.
A genetic variant of the leukocyte phosphatase PTPN22 (R620W) is strongly associated with autoimmune diseases including rheumatoid arthritis (RA). Functional studies on the variant have focussed on lymphocytes, but it is most highly expressed in neutrophils. We have investigated the effects of the variant on neutrophil function in health and in patients with RA. Healthy individuals and patients with RA were genotyped for PTPN22 (R620W) and neutrophils isolated from peripheral blood. Neutrophil adhesion and migration across inflamed endothelium were measured. Calcium (Ca(2+)) release and reactive oxygen species (ROS) production in response to fMLP stimulation were also assessed. Expression of R620W enhanced neutrophil migration through cytokine activated endothelium (non-R620W=24%, R620W=45% migrating cells, p<0.001). Following fMLP stimulation, neutrophils that were heterozygous and homozygous for R620W released significantly more Ca(2+) when compared to non-R620W neutrophils, in healthy individuals and patients with RA. fMLP stimulation, after TNF-α priming, provoked more ROS from neutrophils heterozygous for R620W in patients with RA (non-R620W vs R620W=∼1.75-fold increase) and healthy individuals (non-R620W vs R620W=fourfold increase) and this increase was statistically significant in healthy individuals (p<0.001) but not in patients with RA (p<0.25). Expression of PTPN22 (R620W) enhanced neutrophil effector functions in health and RA, with migration, Ca(2+) release and production of ROS increased. Neutrophils are found in large numbers in the RA joint, and this hyperactivity of R620W cells may directly contribute to the joint damage, as well as to the initiation and perpetuation of the chronic immune-mediated inflammatory processes driving the disease.