Spatio-temporal variation in territory quality and oxidative status: a natural experiment in the Seychelles warbler (Acrocephalus sechellensis).
1. Fluctuations in the quality of the habitat in which an animal lives can have major consequences for its behaviour and physiological state. In poor-quality habitat with low food availability, metabolically intensive foraging activity is likely to result in increased generation of reactive oxygen species, while scarcity of food can lead to a weakening of exogenously derived antioxidant defences. The consequent oxidant/antioxidant imbalance may lead to elevated oxidative stress. 2. Although the link between food availability and oxidative stress has been studied in the laboratory, very little is known about this relationship in the wild. Here, we investigate the association between territory quality (measured through food availability) and oxidative stress in the Seychelles warbler (Acrocephalus sechellensis). 3. Seychelles warblers are insectivorous birds that inhabit a fixed feeding territory year round. Individuals experience profound and rapid local fluctuations in territory quality within these territories, owing to changing patterns of vegetation defoliation resulting from seasonal changes in prevailing wind direction and wind-borne salt spray. 4. As expected, oxidant generation (measured as reactive oxygen metabolites; ROMs) was higher when territory quality was low, but there was no correlation between territory quality and antioxidant capacity (OXY). The negative correlation between territory quality and ROMs was significant between individuals and approached significance within individuals, indicating that the pattern resulted from individual responses to environmental variation. 5. ROMs and OXY levels within individuals were positively correlated, but the relationship between territory quality and ROMs persisted after including OXY as a covariate, implying that oxidative stress occurs in low territory quality conditions. 6. Our results indicate that the oxidative stress balance of an individual is sensitive to relatively short-term changes in territory quality, which may have consequences for the birds' fitness.


Interactive semisupervised learning for microarray analysis.
Microarray technology has generated vast amounts of gene expression data with distinct patterns. Based on the premise that genes of correlated functions tend to exhibit similar expression patterns, various machine learning methods have been applied to capture these specific patterns in microarray data. However, the discrepancy between the rich expression profiles and the limited knowledge of gene functions has been a major hurdle to the understanding of cellular networks. To bridge this gap so as to properly comprehend and interpret expression data, we introduce Relevance Feedback to microarray analysis and propose an interactive learning framework to incorporate the expert knowledge into the decision module. In order to find a good learning method and solve two intrinsic problems in microarray data, high dimensionality and small sample size, we also propose a semisupervised learning algorithm: Kernel Discriminant-EM (KDEM). This algorithm efficiently utilizes a large set of unlabeled data to compensate for the insufficiency of a small set of labeled data and it extends the linear algorithm in Discriminant-EM (DEM) to a kernel algorithm to handle nonlinearly separable data in a lower dimensional space. The Relevance Feedback technique and KDEM together construct an efficient and effective interactive semisupervised learning framework for microarray analysis. Extensive experiments on the yeast cell cycle regulation data set and Plasmodium falciparum red blood cell cycle data set show the promise of this approach.


Obstacles to the integration of abortion into obstetrics and gynecology practice.
Obstetrics and gynecology residents who are trained in family planning and intend to provide abortions after residency often do not ultimately do so. The extent of the professional barriers physicians face trying to integrate abortion into their practice is unknown. In 2006, in-depth interviews were conducted with 30 obstetrician-gynecologists who had graduated 5-10 years earlier from residency programs that included abortion training. Interviews about physicians' experiences with abortion training and practice were coded and analyzed using a grounded theoretical approach. Eighteen physicians had wanted to offer elective abortions after residency, but only three were doing so at the time of the interview. The majority were unable to provide abortions because of formal and informal policies imposed by their private group practices, employers and hospitals, as well as the strain that doing so might put on relationships with superiors and coworkers. Restrictions on abortion provision sometimes were made explicit when new physicians interviewed for a job, but sometimes became apparent only after they had joined a practice or institution. Several physicians mentioned the threat of violence as an obstacle to providing abortions, but few considered this the greatest deterrent. The stigma and ideological contention surrounding abortion manifest themselves in professional environments as barriers to the integration of abortion into medical practice. New physicians often lack the professional support and autonomy necessary to offer abortion services.


Controversies in the management of regional nodes in melanoma.
The surgical management of the regional lymph node basin of melanoma has undergone significant changes in the past 2 decades, most of which have been guided by prospective randomized trials. Historically, routine elective lymph node dissection was recommended for the management of melanoma regardless of clinical nodal involvement. Subsequent randomized trials failed to show a clear benefit for all patients, and sentinel lymph node (SLN) biopsy emerged as an alternative. Although the prognostic value of SLN biopsy in intermediate-thickness melanoma is well accepted, its value for patients with thin and thick lesions is debated. The therapeutic advantage of removing an involved SLN, and the need for a completion lymph node dissection after the identification of a positive SLN, are areas of continued controversy. This article discusses these issues in the management of the regional lymph node basin in patients with melanoma.


Low-temperature collisions between neutral molecules in merged molecular beams.
We have developed an experiment for the investigation of neutral molecular collisions in the gas phase at temperatures as low as 100 mK. These low temperatures are obtained by merging two supersonic expansions, using an electric and a magnetic guide, and by matching the velocities of the beams. Since the energy available for the collisions, or the temperature, is determined only by the relative velocity of the reaction partners this enables the study of chemical processes at very low temperatures without the need to prepare slow molecules in the laboratory frame of reference. This paper describes the method and presents results on the Ne((3)P2)+NH3 Penning ionization.


Precision of dimensional measurements of mandible in different positions on orthopantomogram.
To evaluate the precision of dimensional measurements of the mandible in two different positions on orthopantomogram (OPG) (one with mandibular plane parallel to the floor and the other with Frankfort horizontal plane parallel to the floor) and determine the dimensional reliability. Anatomical landmarks were used to denote points for measurements on mandible as well as OPG and respective measurements made. Magnification was hence calculated and compared with magnification factor (1.2) listed by the manufacturer. Vertical measurements and anterior horizontal and oblique measurements showed minimal magnification. Posterior horizontal and oblique measurements showed increased magnification. The difference in measurements in the two positions and on comparison with the one given by the manufacturer was statistically significant for posterior horizontal measurements but not for posterior oblique. Horizontal measurements crossing the midline were highly magnified and the difference was statistically significant for the two positions and on comparison to the one given by manufacturer. Magnification factor given by the manufacturer is not uniform in all locations and varies with changes in positioning and hence should not be relied upon when accurate measurements are to be made.


Rationale and design of the Canadian Outcomes Registry Late After Tetralogy of Fallot Repair: the CORRELATE study.
Chronic hemodynamically relevant pulmonary regurgitation (PR) resulting in important right ventricular dilation and ventricular dysfunction is commonly seen after tetralogy of Fallot (TOF) repair. Late adverse clinical outcomes, including exercise intolerance, arrhythmias, heart failure and/or death accelerate in the third decade of life and are cause for considerable concern. Timing of pulmonary valve replacement (PVR) to address chronic PR is controversial, particularly in asymptomatic individuals, and effect of PVR on clinical measures has not been determined. Canadian Outcomes Registry Late After Tetralogy of Fallot Repair (CORRELATE) is a prospective, multicentre, Canada-wide cohort study. Candidates will be included if they are ≥ 12 years of age, have had surgically repaired TOF resulting in moderate or severe PR, and are able to undergo cardiovascular magnetic resonance imaging. Enrollment of > 1000 individuals from 15 participating centres (Toronto, Montreal, Quebec City, Sherbrooke, Halifax, Calgary, Edmonton, and Vancouver) is anticipated. Clinical data, health-related quality of life metrics, and adverse outcomes will be entered into a web-based database. A central core lab will analyze all cardiovascular magnetic resonance studies (PR severity, right ventricular volumes, and ventricular function). Major adverse outcomes (sustained ventricular tachycardia and cardiovascular cause of death) will be centrally adjudicated. To the best of our knowledge, CORRELATE will be the first prospective pan-Canadian cohort study of congenital heart disease in children and adults. CORRELATE will uniquely link clinical, imaging, and functional data in those with repaired TOF and important PR, thereby enabling critical evaluation of clinically relevant outcomes in those managed conservatively compared with those referred for PVR.


Platelet transfusions.
Platelets play a pivotal role in hemostasis by forming the initial hemostatic plug and augmenting the formation of the more permanent fibrin thrombus. Thrombocytopenia may lead to bleeding, which can be treated with therapeutic platelet transfusions or prevented with prophylactic transfusions. While many advances have been made in platelet transfusion technology, problems remain. These include the short storage life of liquid-stored platelets and the frequent development of alloantibodies (the refractory state) in patients receiving a number of transfusions. Significant progress, however, is being made in both areas.


Quantitative prediction of formulation-specific food effects and their population variability from in vitro data with the physiologically-based ADAM model: a case study using the BCS/BDDCS Class II drug nifedipine.
Quantitative prediction of food effects (FE) upon drug pharmacokinetics, including population variability, in advance of human trials may help with trial design by optimising the number of subjects and sampling times when a clinical study is warranted or by negating the need for conduct of clinical studies. Classification and rule-based systems such as the BCS and BDDCS and statistical QSARs are widely used to anticipate the nature of FE in early drug development. However, their qualitative rather than quantitative nature makes them less appropriate for assessing the magnitude of FE. Moreover, these approaches are based upon drug properties alone and are not appropriate for estimating potential formulation-specific FE on modified or controlled release products. In contrast, physiologically-based mechanistic models can consider the scope and interplay of a range of physiological changes after food intake and, in combination with appropriate in vitro drug- and formulation-specific data, can make quantitative predictions of formulation-specific FE including the inter-individual variability of such effects. Herein the Advanced Dissolution, Absorption and Metabolism (ADAM) model is applied to the prediction of formulation-specific FE for BCS/BDDCS Class II drug and CYP3A4 substrate nifedipine using as far as possible only in vitro data. Predicted plasma concentration profiles of all three studied formulations under fasted and fed states are within 2-fold of clinically observed profiles. The % prediction error (%PE) in fed-to-fasted ratio of Cmax and AUC were less than 5% for all formulations except for the Cmax of Nifedicron (%PE=-29.6%). This successful case study should help to improve confidence in the use of mechanistic physiologically-based models coupled with in vitro data for the anticipation of FE in advance of in vivo studies. However, it is acknowledged that further studies with drugs/formulations exhibiting a wide range of properties are required to further validate this methodology.


Early Visual Saliency Based on Isolated Optimal Features.
Under fast viewing conditions, the visual system extracts salient and simplified representations of complex visual scenes. Saccadic eye movements optimize such visual analysis through the dynamic sampling of the most informative and salient regions in the scene. However, a general definition of saliency, as well as its role for natural active vision, is still a matter for discussion. Following the general idea that visual saliency may be based on the amount of local information, a recent constrained maximum-entropy model of early vision, applied to natural images, extracts a set of local optimal information-carriers, as candidate salient features. These optimal features proved to be more informative than others in fast vision, when embedded in simplified sketches of natural images. In the present study, for the first time, these features were presented in isolation, to investigate whether they can be visually more salient than other non-optimal features, even in the absence of any meaningful global arrangement (contour, line, etc.). In four psychophysics experiments, fast discriminability of a compound of optimal features (target) in comparison with a similar compound of non-optimal features (distractor) was measured as a function of their number and contrast. Results showed that the saliency predictions from the constrained maximum-entropy model are well verified in the data, even when the optimal features are presented in smaller numbers or at lower contrast. In the eye movements experiment, the target and the distractor compounds were presented in the periphery at different angles. Participants were asked to perform a simple choice-saccade task. Results showed that saccades can select informative optimal features spatially interleaved with non-optimal features even at the shortest latencies. Saccades' choice accuracy and landing position precision improved with SNR. In conclusion, the optimal features predicted by the reference model, turn out to be more salient than others, despite the lack of any clues coming from a global meaningful structure, suggesting that they get preferential treatment during fast image analysis. Also, peripheral fast visual processing of these informative local features is able to guide gaze orientation. We speculate that active vision is efficiently adapted to maximize information in natural visual scenes.


Painful peripheral polyneuropathy after bariatric surgery. Case reports.
The number of patients undergoing bariatric surgery increases every year and the frequency of complications associated with this procedure has been increasing. The main complications reported are nutritional, metabolic, neurological, and psychological/psychiatric. Among the neurological complications, peripheral neuropathies are important due to their high incidence. The objective of this report was to stress the importance of this type of complication, in which the initial manifestation might be neuropathic pain followed or accompanied by motor involvement, and whose prognosis depends on early diagnosis and treatment. The case of three patients were reported, two females and one male, who underwent bariatric surgery (restrictive method - Roux-en-Y gastric bypass) and developed, postoperatively, peripheral neuropathy with neuropathic pain as the initial symptom, with posterior or concomitant motor involvement. Electroneuromyography demonstrated in all three cases a sensitive-motor axonal peripheral neuropathy. In two patients, the fibular nerves were severely affected. In common, they all had an important weight loss in a short period of time before developing neurological symptoms. Neurological complications are one of the most feared complications of bariatric surgeries. Nutritional deficiencies secondary to the surgery are the most important factors in its pathogeny. Nutritional surveillance, avoidance of severe and fast weight loss, and nutritional supplements are fundamental to avoid complications, especially neurological complications.


Painful memories: ritual and the transformation of community trauma.
This paper explores the question of how ritual can heal community pain or trauma. Drawing together recent insights into the ideological nature of healing with a focus on ritual process, I argue that in cases of social violence and healing, ritual's ability to assuage pain is linked to the ways in which it draws pain into the process of reconstructing memory. The argument is illustrated through an examination of how the Betsimisaraka of east Madagascar used rituals of cattle sacrifice to transform the pain they experienced during an anticolonial rebellion that took place in 1947.


Environmental enrichment alters protein expression as well as the proteomic response to cocaine in rat nucleus accumbens.
Prior research demonstrated that environmental enrichment creates individual differences in behavior leading to a protective addiction phenotype in rats. Understanding the mechanisms underlying this phenotype will guide selection of targets for much-needed novel pharmacotherapeutics. The current study investigates differences in proteome expression in the nucleus accumbens of enriched and isolated rats and the proteomic response to cocaine self-administration using a liquid chromatography mass spectrometry (LCMS) technique to quantify 1917 proteins. Results of complementary Ingenuity Pathways Analyses (IPA) and gene set enrichment analyses (GSEA), both performed using protein quantitative data, demonstrate that cocaine increases vesicular transporters for dopamine and glutamate as well as increasing proteins in the RhoA pathway. Further, cocaine regulates proteins related to ERK, CREB and AKT signaling. Environmental enrichment altered expression of a large number of proteins implicated in a diverse number of neuronal functions (e.g., energy production, mRNA splicing, and ubiquitination), molecular cascades (e.g., protein kinases), psychiatric disorders (e.g., mood disorders), and neurodegenerative diseases (e.g., Huntington's and Alzheimer's diseases). Upregulation of energy metabolism components in EC rats was verified using RNA sequencing. Most of the biological functions and pathways listed above were also identified in the Cocaine X Enrichment interaction analysis, providing clear evidence that enriched and isolated rats respond quite differently to cocaine exposure. The overall impression of the current results is that enriched saline-administering rats have a unique proteomic complement compared to enriched cocaine-administering rats as well as saline and cocaine-taking isolated rats. These results identify possible mechanisms of the protective phenotype and provide fertile soil for developing novel pharmacotherapeutics. Proteomics data are available via ProteomeXchange with identifier PXD000990.


Innate lymphoid cells are reduced in pregnant HIV positive women and are associated with preterm birth.
Preterm birth is the leading cause of neonatal and child mortality worldwide. Globally, 1.4 million pregnant women are estimated to be living with HIV/AIDS, the majority of whom live in sub-Saharan Africa. Maternal HIV infection and antiretroviral treatment (ART) have been associated with increased rates of preterm birth, but the underlying mechanisms remain unknown. Acute HIV infection is associated with a rapid depletion of all three subsets of innate lymphoid cells (ILCs), ILC1s, ILC2s and ILC3s, which is not reversed by ART. ILCs have been found at the maternal-fetal interface and we therefore investigated the potential association between maternal HIV infection, peripheral ILC frequencies and preterm birth. In our study of pregnant South African women with accurately dated pregnancies, we show that maternal HIV infection is associated with reduced levels of all three ILC subsets. Preterm birth was also associated with lower levels of all three ILC subsets in early pregnancy. ILC frequencies were lowest in HIV positive women who experienced preterm birth. Moreover, ILC levels were reduced in pregnancies resulting in spontaneous onset of preterm labour and in extreme preterm birth (< 28 weeks gestation). Our findings suggest that reduced ILC frequencies may be a link between maternal HIV infection and preterm birth. In addition, ILC frequencies in early pregnancy may serve as predictive biomarkers for women who are at risk of delivering preterm.


Effects of adding fluids to solid foods on muscle activity and number of chewing cycles.
The production of a sufficient amount of saliva is indispensable for good chewing. In the present study, we examined the hypothesis that adding fluid to a food will facilitate the chewing process, especially for dry foods. The effect might be larger for subjects with relatively low salivary flow rates. Furthermore, adding fluids that contain mucins or alpha-amylase may have a larger facilitating effect on mastication than the addition of water alone. Twenty subjects chewed on melba toast, breakfast cake, carrot, peanut, and Gouda cheese. In addition, they chewed on these foods after different volumes of water, artificial saliva containing mucins, or a solution of alpha-amylase had been added. Muscle activity and number of chewing strokes until swallowing were measured. The salivary flow rates of the subjects were also determined. Adding fluid to the food significantly reduced the number of chewing cycles and total muscular work (i.e. the integrated surface electromyograpy of masseter and temporalis muscles measured bilaterally, summed for all chewing cycles) until swallowing for all foods, except carrot. The largest effects were observed for melba and cake, which are dry products requiring sufficient saliva to form a coherent bolus safe for swallowing. More facilitation of the chewing process was observed after adding fluid to breakfast cake for subjects with relatively low salivary flow rates. The type of fluid had no significant effect on the chewing process.


Thoracic, abdominal, and vertebral actinomycosis. Diagnosis and long-term therapy in three dogs.
The clinical, laboratory, radiographic, and histologic features and the response to therapy in three dogs with actinomycosis are reported. One dog (dog 1) had a 12-cm nonresectable mass extending from the ventrolateral chest wall into the left ventricular myocardium. Another dog (dog 2) had a diffuse peritonitis with "sulfur granules" and two large masses. One of these masses was nonresectable involving adjacent abdominal structures. A third dog (dog 3) had a subvertebral mass at T1-3 producing quadraplegia. Two dogs had periosteal reactions involving adjacent sternebrae (dog 1) or ribs and vertebral bodies (dog 3) that are characteristic of Actinomyces spp infections. In dogs 1 and 2 the diagnosis was based on the morphologic and tinctorial properties of free sulfur granules and/or tissue granules. Culture results were variable. Tissue from dog 1 yielded no growth, while polymicrobial infections, which included Actinomyces spp, were identified in dogs 2 and 3. Actinomyces odontolyticus was isolated from dog 3. Although the actinomycotic granulomas were either not excised or only partially excised from dogs 1 and 2, both animals were cured by the oral administration of high doses of penicillin G for 19 and 6 months, respectively. Dog 3 responded dramatically to the same antibiotic therapy given for 5 months. However, within 4 months of discontinuing treatment an abscess and draining fistulous tracts developed in the left axillary region. Two surgical fistulectomies and additional penicillin therapy were required to cure this animal. These cases and the current veterinary and human literature on actinomycosis are used to propose a rational approach to the treatment of actinomycosis in the dog.


Community-based participatory research: development of an emergency department-based youth violence intervention using concept mapping.
Emergency departments (EDs) see a high number of youths injured by violence. In Ontario, the most common cause of injury for youths visiting EDs is assault. Secondary prevention strategies using the teachable moment (i.e., events that can lead individuals to make positive changes in their lives) are ideal for use by clinicians. An opportunity exists to take advantage of the teachable moment in the ED in an effort to prevent future occurrences of injury in at-risk youths. However, little is known about perceptions of youths, parents, and community organizations about such interventions in EDs. The aims of this study were to engage youths, parents, and frontline community workers in conceptualizing a hospital-based violence prevention intervention and to identify outcomes relevant to the community. Concept mapping is an innovative, mixed-method research approach. It combines structured qualitative processes such as brainstorming and group sorting, with various statistical analyses such as multidimensional scaling and hierarchical clustering, to develop a conceptual framework, and allows for an objective presentation of qualitative data. Concept mapping involves multiple structured steps: 1) brainstorming, 2) sorting, 3) rating, and 4) interpretation. For this study, the first three steps occurred online, and the fourth step occurred during a community meeting. Over 90 participants were involved, including youths, parents, and community youth workers. A two-dimensional point map was created and clusters formed to create a visual display of participant ideas on an ED-based youth violence prevention intervention. Issues related to youth violence prevention that were rated of highest importance and most realistic for hospital involvement included mentorship, the development of youth support groups in the hospital, training doctors and nurses to ask questions about the violent event, and treating youth with respect. Small-group discussions on the various clusters developed job descriptions, a list of essential services, and suggestions on ways to create a more youth-friendly environment in the hospital. A large-group discussion revealed outcomes that participants felt should be measured to determine the success of an intervention program. This study has been the springboard for the development of an ED-based youth violence intervention that is supported by the community and affected youth. Using information generated by youth that is grounded in their experience through participatory research methods is feasible for the development of successful and meaningful youth violence prevention interventions.


Immunocytochemical markers revealing retinal and pineal but not hypothalamic photoreceptor systems in the Japanese quail.
The retinal proteins opsin, alpha-transducin, S-antigen and interstitial retinol-binding protein (IRBP) are essential for the processes of vision. By use of immunocytochemistry we have employed antibodies directed against these "photoreceptor proteins" in an attempt to identify the photoreceptor systems (retina, pineal and deep brain) of the Japanese quail. Opsin immunostaining was identified within many outer (basal portion) and inner segments of retinal photoreceptor cells and limited numbers of photoreceptor perikarya. Opsin immunostaining was also demonstrated in limited numbers of pinealocytes with all parts of these cells being immunoreactive. These results differ from previous observations. In contrast to the results obtained with the antibody against opsin, S-antigen and alpha-transducin immunostaining was seen throughout the entire outer segments and many photoreceptor perikarya of the retina, in the pineal organ immunostaining was seen in numerous pinealocytes in all follicles. These results conform to previous findings in birds. In addition, IRBP has been demonstrated for the first time in the avian retina and pineal organ. These findings underline the structural and functional similarities between the retina and pineal organ and provide additional support for a photoreceptive role of the avian pineal. No specific staining was detected in any other region of the brain in the Japanese quail; the hypothalamic photoreceptors of birds remain unidentified.


An atomistic mechanism study of GaN step-flow growth in vicinal m-plane orientations.
Elucidation of homoepitaxial growth mechanisms on vicinal non-polar surfaces of GaN is highly important for gaining an understanding of and control thin film surface morphology and properties. Using first-principles calculations, we study the step-flow growth in m-plane GaN based on atomic row nucleation and kink propagation kinetics. Ga-N dimer adsorption onto the m-plane is energetically more favorable than that of Ga and N isolated adatoms. Therefore, we have treated the dimers as the dominant growth species attached to the step edges. By calculating the free energies of sequentially attached Ga-N dimers, we have elucidated that the a-step edge kink growth proceeds by parallel attachment rather than by across the step edge approach. We found a series of favorable configurations of kink propagation and calculated the free energy and nucleation barriers for kink evolution on five types of step edges (a, +c, -c, +a + c, and -a - c). By changing the chemical potential μGa and the excess chemical potential Δμ, the growth velocities at the five types of edges are controlled by the corresponding kink pair nucleation barrier E* in their free energy profiles. To explore the kink-flow growth instability observed at different Ga/N flux ratios, calculations of kink pairs on the incompact -c and +c-step edges are further performed to study their formation energies. Variations of these step edge morphologies with a tuned chemical environment are consistent with previous experimental observations, including stable diagonal ±a ± c-direction steps. Our work provides a first-principles approach to explore step growth and surface morphology of the vicinal m-plane GaN, which is applicable to analyze and control the step-flow growth of other binary thin films.


A Systematic Review of the Efficacy and Safety of Microneedling in the Treatment of Melasma.
Melasma is an acquired disorder of hyperpigmentation that is often recalcitrant to current therapies. Microneedling is used to treat scars, striae, and rhytides and has a relatively low risk of post-treatment dyspigmentation. Several studies have examined its use in melasma. To review the published evidence on the efficacy and safety of microneedling in the treatment of melasma. A systematic review was performed. A meta-analysis could not be performed because of methodological differences across studies and data heterogeneity. Eight studies were included for analysis. Most studies assessed the utility of microneedling in combination with other topical therapies and detected some success. However, microneedling-mediated transdermal delivery of medications is not superior to microinjections of medications. There is less evidence supporting the use of microneedling as monotherapy. Microneedling, when used with a 1064-nm Q-switched Nd:YAG laser, may provide additional benefit, although with a risk of post-treatment dyspigmentation. Based on low-quality evidence, microneedling may play a role in the treatment of melasma, with the mechanism of action likely being the facilitation of delivery of topical therapies to the epidermis and dermis, and one ancillary benefit of this approach being the very low risk of postinflammatory hyperpigmentation.


Household poverty, schooling, stigma and quality of life in adolescents with epilepsy in rural Uganda.
Epilepsy remains a leading chronic neurological disorder in Low- and Middle-Income Countries. In Uganda, the highest burden is among young rural people. We aimed to; (i) describe socio-economic status (including schooling), and household poverty in adolescents living with epilepsy (ALE) compared to unaffected counterparts in the same communities and (ii) determine the factors associated with the overall quality of life (QoL). This was a cross-sectional survey nested within a larger study of ALE compared to age-matched healthy community children in Uganda. Between Sept 2016 to Sept 2017, 154 ALE and 154 healthy community controls were consecutively recruited. Adolescents recruited were frequency and age-matched based on age categories 10-14 and 15-19 years. Clinical history and standardized assessments were conducted. One control participant had incomplete assessment and was excluded. The primary outcome was overall QoL and key variables assessed were schooling status and household poverty. Descriptive and multivariable linear regression analysis were conducted for independent associations with overall QoL. Mean (SD) age at seizure onset was 8.8 (3.9) years and median (IQR) monthly seizure burden was 2 (1-4). Epilepsy was associated with living in homes with high household poverty; 95/154 (61.7%) ALE lived in the poorest homes compared to 68/153 (44.5%) of the healthy adolescents, p = 0.001. Nearly two-thirds of ALE had dropped out of school and only 48/154 (31.2%) were currently attending school compared to 136/153 (88.9%) of healthy controls, p < 0.001. QoL was lowest among ALE who never attended school (p < 0.001), with primary education (p = 0.006) compared to those with at least secondary education. Stigma scores [mean(SD)] were highest among ALE in the poorest [69.1(34.6)], and wealthy [70.2(32.2)] quintiles compared to their counterparts in poorer [61.8(31.7)], medium [68.0(32.7)] and wealthiest [61.5(33.3)] quintiles, though not statistically significant (p = 0.75). After adjusting for covariates, ALE currently attending school had higher overall QoL compared to their counterparts who never attended school (β = 4.20, 95%CI: 0.90,7.49, p = 0.013). QoL scores were higher among ALE with ≥secondary education than those with no or primary education (β = 10.69, 95%CI: 1.65, 19.72). ALE in this rural area are from the poorest households, are more likely to drop out of school and have the lowest QoL. Those with poorer seizure control are most affected. ALE should be included among vulnerable population groups and in addition to schooling, strategies for seizure control and addressing the epilepsy treatment gap in affected homes should be specifically targeted in state poverty eradication programs.


Integrating genomes.
As genomic sequencing projects attempt ever more ambitious integration of genetic, molecular, and phenotypic information, a specialization of genomics has emerged, embodied in the subdiscipline of computational genomics. Models inherited from population genetics, phylogenetics, and human disease genetics merge with those from graph theory, statistics, signal processing, and computer science to provide a rich quantitative foundation for genomics that can only be realized with the aid of a computer. Unleashed on a rapidly increasing sample of the planet's 10(30) organisms, these analyses will have an impact on diverse fields of science while providing an extraordinary new window into the story of life.


Randomized controlled non-inferiority trial of a telehealth treatment for chronic stuttering: the Camperdown Program.
Although there are treatments that can alleviate stuttering in adults for clinically significant periods, in Australia there are barriers to the accessibility and availability of best-practice treatment. This parallel group, non-inferiority randomized controlled trial with multiple blinded outcome assessments investigated whether telehealth delivery of the Camperdown Program provides a non-inferior alternative to face-to-face treatment for adults who stutter. Forty participants who presented to a university speech clinic were randomized: 20 to the telehealth arm and 20 to the face-to-face arm. Exclusion criteria were age younger than 18 years, frequency of stuttering less than 2% of syllables stuttered and previous speech-restructuring treatment within the past 12 months. The Camperdown Program for adults who stutter was the intervention. Primary outcome measures were frequency of stuttering measured in per cent syllables stuttered (%SS) before treatment and at 9 months post-randomization and efficiency, measured by counting the number of speech pathologist contact hours used by each participant. Intention to treat analysis was conducted using last observation carried forward. Secondary outcome measures were speech naturalness, self-reported stuttering severity, and treatment satisfaction. There was no statistically or clinically significant difference in %SS between the two groups at 9 months post-randomization. Analysis of covariance adjusting for baseline %SS showed telehealth had 0.8% absolute lower per cent syllables stuttered than face-to-face. There were also no differences in %SS between groups immediately post-treatment, or at 6 months and 12 months post-treatment (p = 0.9). In the second primary outcome measure, the telehealth group used statistically less contact time (221 min) on average than the face-to-face group (95% confidence interval = - 387 to - 56 min, p = 0.01). The results provide evidence to support the use of the Camperdown Program delivered by telehealth as an alternate to the face-to-face treatment delivery of this programme for adults who stutter. Such a model will increase accessibility to this evidence-based treatment for adults currently isolated from treatment services.


Use of HIV-1 p24 as a sensitive, precise and inexpensive marker for infection, disease progression and treatment failure.
HIV RNA is an acknowledged marker of disease activity and predictive of progression, while the p24 antigen is considered unsuitable. This is at odds with the fact that viral pathogenesis is usually mediated by proteins. One might expect that p24, if analyzed properly, might even be superior to RNA. This hypothesis was investigated in clinical studies using a sensitive and precise p24 test (heat-mediated immune complex dissociation with signal amplification-boosted ELISA). This test was as sensitive and specific as the polymerase chain reaction (PCR) for viral RNA (200-400 copy detection), an overall better predictor of CD4 decline and survival, while RNA prevailed in predicting AIDS. The lower costs of p24 testing also permit a closer monitoring of patients with an earlier detection of anti-retroviral treatment failures.


Factors predicting mortality in rural elderly hospitalized for pneumonia.
To identify predictors of mortality, the records of 133 elderly patients with pneumonia admitted to a small rural midwestern hospital were examined using a retrospective cohort design. All recorded clinical information available to the patient's physician within the first hours of admission was reviewed. Twenty-one (15.8%) patients died during the hospitalization. Patients with preexisting coronary heart disease, dementia, urinary incontinence, and impaired mobility were more likely to die. Impaired mental status, absence of fever, rapid respiratory rate, hypotension, cyanosis, and diffuse abnormalities on chest examination were also associated with mortality. Logistic regression analysis revealed five predictive indicators of mortality: impaired level of consciousness (odds ratio [OR] = 11.3), tachypnea (OR = 10.8), temperature lower than normal (OR = 14.2), white cell count higher than 20 X 10(9)/L (20,000 mm-3) (OR = 12.2), and cyanosis (OR = 8.6). A risk score based on this regression model demonstrated that 1 of 95 patients with a score lower than 3 (1%), 7 of 22 with a score of 3 (32%), and 13 of 15 patients with a score higher than 3 (87%) died during their hospitalization. The validity of this risk-scoring system was confirmed in another sample of 40 patients. Studies such as this may be useful in identifying information of important prognostic value that enables physicians, patients, and family members to make more effective decisions.


An experimental and theoretical study of the gas phase kinetics of atomic chlorine reactions with CH3NH2, (CH3)2NH, and (CH3)3N.
The rate coefficients for the reactions of Cl((2)PJ) with methylamine (R1), dimethylamine (R2) and trimethylamine (R3) have been measured using the laser flash photolysis - resonance fluorescence technique as a function of temperature (274-435 K) and pressure (25-400 Torr N2). The experimental data are well-represented by the following temperature- and pressure-independent rate coefficients (10(10) × k/cm(3) molecule(-1) s(-1)): kR1 = 2.90 ± 0.44, kR2 = 3.89 ± 0.58, kR3 = 3.68 ± 0.55; the uncertainties are estimates of accuracy at the 95% confidence level. Potential energy surfaces (PES) for the reactions have been characterized at the MP2/cc-pVTZ level and improved single point energies of stationary points obtained in CCSD(T)-F12a calculations. The PES for all reactions are characterized by the formation of pre and post reaction complexes and submerged barriers. Rate coefficients for the reactions were calculated as a function of temperature and pressure using a master equation model based on the coupled cluster theory results. The calculated rate coefficients are in good agreement with experiment; the overall rate coefficients are relatively insensitive to variations of the barrier heights within typical chemical accuracy, but the predicted branching ratios vary significantly. The inclusion of tunnelling has no effect.


Global rating of change: perspectives of patients with lumbar impairments and of their physical therapists.
Background: Global rating of change (GROC) scores provide a means of measuring patients' self-perceived change in health status over time. Objectives: The purposes of the study were to examine agreement of GROC ratings between the patient (GROCp) and the treating therapist (GROCt), the impact on the minimal clinically important difference (MCID) estimates by reporting resources, and the relationships between the subjective GROC scores and objective outcome measures. Methods: Data from 52,767 patients with orthopedic lumbar spine impairments treated in 623 outpatient rehabilitation clinics in 34 states (United States) were analyzed. Patient's functional status (FS) before and after the treatment was measured using the Lumbar Computerized Adaptive Testing (LCAT) survey (0-100 scale). The GROC ratings were obtained from both the patients and their caring therapists at discharge. Results: On average, GROCp (mean [(SD] = 3.1 [4.1]) was 1.9 points lower than GROCt (mean [SD] = 5.0 [1.9]). There were negligible differences between MCID estimates derived from the patients' GROC ratings (MCID cutoff = 9.2) and from the therapists' GROC ratings (MCID cutoff = 9.5). While only analyzing patient data whose GROCp ≥ 0, the intraclass correlation coefficient between GROCp and GROCt was 0.61, the GROCp correlated with FS at intake at r = 0.12, with FS at discharge at r = 0.56, and with FS change scores at r = 0.47. Conclusions: Therapists tended to report higher GROC ratings than their patients did but there were negligible differences between MCID estimates derived from GROCp and that derived from GROCt.


Rationale and study design of MOTION: A phase 4, prospective, single-arm, open-label study to measure outcomes in patients with pulmonary arterial hypertension not on active treatment.
In clinical trials of treatments for pulmonary arterial hypertension (PAH), objective measures, such as 6-min walk distance (6MWD) are limited in their ability to characterize the impact of PAH therapy from a patient's perspective. Few clinical studies have evaluated the primary effects of pharmacologic treatment on patient-reported outcomes, such as symptoms, health-related quality of life (HRQoL), and productivity. MOTION (NCT02191137) is a prospective, multicenter, single-arm, open-label, phase 4 trial designed to assess whether riociguat monotherapy will improve patient-reported outcomes in patients with PAH in the United States who are not currently on treatment. Following a screening period of up to 14 days, eligible subjects will receive riociguat (0.5-2.5 mg TID) during a 10-week titration phase and a 14-week maintenance phase. The primary endpoint is change from baseline in the Living with Pulmonary Hypertension (LPH) questionnaire, a disease-specific HRQoL measure, after 24 weeks of riociguat treatment. The Short Form-12 Health Survey (SF-12) and the Work Limitations Questionnaire 8 (WLQ-8) will also be utilized to assess patient-reported outcomes. Other variables include change from baseline in World Health Organization functional class, 6MWD, and modified Borg Dyspnea Index. In addition, accelerator band activity will be validated against the 6MWD test. Safety will also be assessed. The MOTION trial will provide information on the effect of riociguat on patient-reported outcomes in PAH patients in the United States who are not currently on active treatment through the use of disease-specific and generic HRQoL measures (LPH and SF-12) and a measure of worker productivity (WLQ-8).


Deguelin induced differentiation of mutated NPM1 acute myeloid leukemia in vivo and in vitro.
Nucleophosmin (NPM1), a restricted nucleolar localization protein, shuttles between the nucleus and the cytoplasm. Mutated (Mt)-NPM1 protein, which has aberrant cytoplasmic dislocation of nucleophosmin, occurs in approximately one-third of acute myeloid leukemia cases. Deguelin, a rotenoid isolated from several plant species, is a strong antitumor agent. NOD/SCID mice xenografted with human Mt-NPM1 OCI/AML3 cell lines served as in-vivo models. Wright-Giemsa staining and flow cytometry analysis were used for differentiation assays. Associated molecular events were assessed by western blot and histological analyses. Kaplan-Meier estimates were used to calculate survival. Deguelin toxicity in mice was assessed by immunohistochemistry staining and serum markers. Clinical samples were differentiated by flow cytometry analysis. Deguelin induced differentiation by downregulating the Mt-NPM1 protein levels, which was accompanied by a decrease in SIRT1, p21, and HDAC1 and an increase in CEBPβ and granulocyte colony-stimulating factor receptor protein expression levels. A low-deguelin dose prolonged survival compared with the control group, and there were no apparent lesions to the brain, liver, heart, and kidney in vivo. In clinical samples, deguelin induced the differentiation of fresh blasts with Mt-NPM1 protein, but not with the wild-type NPM1 protein. Taken together, these findings further provide new evidence that the Mt-NPM1 protein plays an important role in inducing differentiation in vivo and in vitro. Mutated NPM1 protein may be a therapeutic target of deguelin in acute myeloid leukemia with the NPM1 mutation.


Activation of peroxisome proliferator-activated receptor delta induces fatty acid beta-oxidation in skeletal muscle and attenuates metabolic syndrome.
In this study, we defined the role of peroxisome proliferator-activated receptor beta/delta (PPARdelta) in metabolic homeostasis by using subtype selective agonists. Analysis of rat L6 myotubes treated with the PPARdelta subtype-selective agonist, GW501516, by the Affymetrix oligonucleotide microarrays revealed that PPARdelta controls fatty acid oxidation by regulating genes involved in fatty acid transport, beta-oxidation, and mitochondrial respiration. Similar PPARdelta-mediated gene activation was observed in the skeletal muscle of GW501516-treated mice. Accordingly, GW501516 treatment induced fatty acid beta-oxidation in L6 myotubes as well as in mouse skeletal muscles. Administration of GW501516 to mice fed a high-fat diet ameliorated diet-induced obesity and insulin resistance, an effect accompanied by enhanced metabolic rate and fatty acid beta-oxidation, proliferation of mitochondria, and a marked reduction of lipid droplets in skeletal muscles. Despite a modest body weight change relative to vehicle-treated mice, GW501516 treatment also markedly improved diabetes as revealed by the decrease in plasma glucose and blood insulin levels in genetically obese ob/ob mice. These data suggest that PPARdelta is pivotal to control the program for fatty acid oxidation in the skeletal muscle, thereby ameliorating obesity and insulin resistance through its activation in obese animals.


The relationship of anticardiolipin antibodies to disease activity in systemic lupus erythematosus.
Sera from 124 blood donors, 60 rheumatoid arthritis (RA) and 57 SLE patients were measured for anticardiolipin antibodies by ELISA. Significantly raised IgG (aCLG) and IgM (aCLM) anticardiolipin antibody levels were found in RA and SLE (p less than 0.0005). However, in SLE both aCLG and aCLM levels were significantly higher than in RA (p less than 0.0025). We then conducted a transectional study to evaluate aCL levels and disease activity in SLE. There was a good positive predictive value (70%) between aCL and overall disease activity, but not for individual systems. A strong association between aCL and renal involvement irrespective of activity was also found (80%). Nine SLE patients fulfilled both the clinical and serological criteria for the antiphospholipid syndrome (APS) and a further 18 patients fulfilled the serological criteria for APS. Results indicate that aCL levels are of value in predicting overall disease activity in SLE and in monitoring those patients who fulfil or partially fulfil the criteria for APS.


What can individuals do to reduce personal health risks from air pollution?
In many areas of the world, concentrations of ambient air pollutants exceed levels associated with increased risk of acute and chronic health problems. While effective policies to reduce emissions at their sources are clearly preferable, some evidence supports the effectiveness of individual actions to reduce exposure and health risks. Personal exposure to ambient air pollution can be reduced on high air pollution days by staying indoors, reducing outdoor air infiltration to indoors, cleaning indoor air with air filters, and limiting physical exertion, especially outdoors and near air pollution sources. Limited evidence suggests that the use of respirators may be effective in some circumstances. Awareness of air pollution levels is facilitated by a growing number of public air quality alert systems. Avoiding exposure to air pollutants is especially important for susceptible individuals with chronic cardiovascular or pulmonary disease, children, and the elderly. Research on mechanisms underlying the adverse health effects of air pollution have suggested potential pharmaceutical or chemopreventive interventions, such as antioxidant or antithrombotic agents, but in the absence of data on health outcomes, no sound recommendations can be made for primary prevention. Health care providers and their patients should carefully consider individual circumstances related to outdoor and indoor air pollutant exposure levels and susceptibility to those air pollutants when deciding on a course of action to reduce personal exposure and health risks from ambient air pollutants. Careful consideration is especially warranted when interventions may have unintended negative consequences, such as when efforts to avoid exposure to air pollutants lead to reduced physical activity or when there is evidence that dietary supplements, such as antioxidants, have potential adverse health effects. These potential complications of partially effective personal interventions to reduce exposure or risk highlight the primary importance of reducing emissions of air pollutants at their sources.


Routine RNA sequencing of formalin-fixed paraffin-embedded specimens in neuropathology diagnostics identifies diagnostically and therapeutically relevant gene fusions.
Molecular markers have become pivotal in brain tumor diagnostics. Mutational analyses by targeted next-generation sequencing of DNA and array-based DNA methylation assessment with copy number analyses are increasingly being used in routine diagnostics. However, the broad variety of gene fusions occurring in brain tumors is marginally covered by these technologies and often only assessed by targeted assays. Here, we assessed the feasibility and clinical value of investigating gene fusions in formalin-fixed paraffin-embedded (FFPE) tumor tissues by next-generation mRNA sequencing in a routine diagnostic setting. After establishment and optimization of a workflow applicable in a routine setting, prospective diagnostic application in a neuropathology department for 26 months yielded relevant fusions in 66 out of 101 (65%) analyzed cases. In 43 (43%) cases, the fusions were of decisive diagnostic relevance and in 40 (40%) cases the fusion genes rendered a druggable target. A major strength of this approach was its ability to detect fusions beyond the canonical alterations for a given entity, and the unbiased search for any fusion event in cases with uncertain diagnosis and, thus, uncertain spectrum of expected fusions. This included both rare variants of established fusions which had evaded prior targeted analyses as well as the detection of previously unreported fusion events. While the impact of fusion detection on diagnostics is highly relevant, it is especially the detection of "druggable" fusions which will most likely provide direct benefit to the patients. The wider application of this approach for unbiased fusion identification therefore promises to be a major advance in identifying alterations with immediate impact on patient care.


Activating E2Fs mediate transcriptional regulation of human E2F6 repressor.
E2F6 is believed to repress E2F-responsive genes and therefore serve a role in cell cycle regulation. Analysis of the human E2F6 promoter region revealed the presence of two putative E2F binding sites, both of which were found to be functionally critical because deletion or mutations of these sites abolished promoter activity. Ectopic expression of E2F1 protein was found to increase E2F6 mRNA levels and significantly upregulate E2F6 promoter activity. Deletion or mutation of the putative E2F binding sites nullified the effects of E2F1 on the E2F6 promoter activity. Studies on the temporal induction of E2F family members demonstrated that the activating E2Fs, and most notably E2F1, were upregulated before E2F6 during cell cycle progression at the G1/S phase, and this coincided with the time course of induction experienced by the E2F6 promoter during the course of the cell cycle. EMSAs indicated the specific binding of nuclear complexes to the E2F6 promoter that contained E2F1-related species whose binding was specifically competed by the consensus E2F binding site. Chromatin immunoprecipitation assays with anti-E2Fs demonstrated the association of E2F family members with the E2F6 promoter in vivo. These data indicate that the expression of the E2F6 repressor is influenced at the transcriptional level by E2F family members and suggest that interplay among these transcriptional regulators, especially E2F1, may be critical for cell cycle regulation.


Perinatal iron deficiency alters the neurochemical profile of the developing rat hippocampus.
Cognitive deficits in human infants at risk for gestationally acquired perinatal iron deficiency suggest involvement of the developing hippocampus. To understand the plausible biological explanations for hippocampal injury in perinatal iron deficiency, a neurochemical profile of 16 metabolites in the iron-deficient rat hippocampus was evaluated longitudinally by 1H NMR spectroscopy at 9.4 T. Metabolites were quantified from an 11-24 microL volume centered in the hippocampus in 18 iron-deficient and 16 iron-sufficient rats on postnatal day (PD) 7, PD10, PD14, PD21 and PD28. Perinatal iron deficiency was induced by feeding the pregnant dam an iron-deficient diet from gestational d 3 to PD7. The brain iron concentration of the iron-deficient group was 60% lower on PD7 and 19% lower on PD28 (P < 0.001 each). The concentration of 12 of the 16 measured metabolites changed over time between PD7 and PD28 in both groups (P < 0.001 each). Compared with the iron-sufficient group, phosphocreatine, glutamate, N-acetylaspartate, aspartate, gamma-aminobutyric acid, phosphorylethanolamine and taurine concentrations, and the phosphocreatine/creatine ratio were elevated in the iron-deficient group (P < 0.02 each). These neurochemical alterations suggest persistent changes in resting energy status, neurotransmission and myelination in perinatal iron deficiency. An altered neurochemical profile of the developing hippocampus may underlie some of the cognitive deficits observed in human infants with perinatal iron deficiency.


A methodology to assess the intrinsic discriminative ability of a distance function and its interplay with clustering algorithms for microarray data analysis.
Clustering is one of the most well known activities in scientific investigation and the object of research in many disciplines, ranging from statistics to computer science. Following Handl et al., it can be summarized as a three step process: (1) choice of a distance function; (2) choice of a clustering algorithm; (3) choice of a validation method. Although such a purist approach to clustering is hardly seen in many areas of science, genomic data require that level of attention, if inferences made from cluster analysis have to be of some relevance to biomedical research. A procedure is proposed for the assessment of the discriminative ability of a distance function. That is, the evaluation of the ability of a distance function to capture structure in a dataset. It is based on the introduction of a new external validation index, referred to as Balanced Misclassification Index (BMI, for short) and of a nontrivial modification of the well known Receiver Operating Curve (ROC, for short), which we refer to as Corrected ROC (CROC, for short). The main results are: (a) a quantitative and qualitative method to describe the intrinsic separation ability of a distance; (b) a quantitative method to assess the performance of a clustering algorithm in conjunction with the intrinsic separation ability of a distance function. The proposed procedure is more informative than the ones available in the literature due to the adopted tools. Indeed, the first one allows to map distances and clustering solutions as graphical objects on a plane, and gives information about the bias of the clustering algorithm with respect to a distance. The second tool is a new external validity index which shows similar performances with respect to the state of the art, but with more flexibility, allowing for a broader spectrum of applications. In fact, it allows not only to quantify the merit of each clustering solution but also to quantify the agglomerative or divisive errors due to the algorithm. The new methodology has been used to experimentally study three popular distance functions, namely, Euclidean distance d2, Pearson correlation dr and mutual information dMI. Based on the results of the experiments, we have that the Euclidean and Pearson correlation distances have a good intrinsic discrimination ability. Conversely, the mutual information distance does not seem to offer the same flexibility and versatility as the other two distances. Apparently, that is due to well known problems in its estimation. since it requires that a dataset must have a substantial number of features to be reliable. Nevertheless, taking into account such a fact, together with results presented in Priness et al., one receives an indication that dMI may be superior to the other distances considered in this study only in conjunction with clustering algorithms specifically designed for its use. In addition, it results that K-means, Average Link, and Complete link clustering algorithms are in most cases able to improve the discriminative ability of the distances considered in this study with respect to clustering. The methodology has a range of applicability that goes well beyond microarray data since it is independent of the nature of the input data. The only requirement is that the input data must have the same format of a "feature matrix". In particular it can be used to cluster ChIP-seq data.


Nanophotonic integration in state-of-the-art CMOS foundries.
We demonstrate a monolithic photonic integration platform that leverages the existing state-of-the-art CMOS foundry infrastructure. In our approach, proven XeF2 post-processing technology and compliance with electronic foundry process flows eliminate the need for specialized substrates or wafer bonding. This approach enables intimate integration of large numbers of nanophotonic devices alongside high-density, high-performance transistors at low initial and incremental cost. We demonstrate this platform by presenting grating-coupled, microring-resonator filter banks fabricated in an unmodified 28 nm bulk-CMOS process by sharing a mask set with standard electronic projects. The lithographic fidelity of this process enables the high-throughput fabrication of second-order, wavelength-division-multiplexing (WDM) filter banks that achieve low insertion loss without post-fabrication trimming.


Metabolic engineering of carbon and redox flow in the production of small organic acids.
The review describes efforts toward metabolic engineering of production of organic acids. One aspect of the strategy involves the generation of an appropriate amount and type of reduced cofactor needed for the designed pathway. The ability to capture reducing power in the proper form, NADH or NADPH for the biosynthetic reactions leading to the organic acid, requires specific attention in designing the host and also depends on the feedstock used and cell energetic requirements for efficient metabolism during production. Recent work on the formation and commercial uses of a number of small mono- and diacids is discussed with redox differences, major biosynthetic precursors and engineering strategies outlined. Specific attention is given to those acids that are used in balancing cell redox or providing reduction equivalents for the cell, such as formate, which can be used in conjunction with metabolic engineering of other products to improve yields. Since a number of widely studied acids derived from oxaloacetate as an important precursor, several of these acids are covered with the general strategies and particular components summarized, including succinate, fumarate and malate. Since malate and fumarate are less reduced than succinate, the availability of reduction equivalents and level of aerobiosis are important parameters in optimizing production of these compounds in various hosts. Several other more oxidized acids are also discussed as in some cases, they may be desired products or their formation is minimized to afford higher yields of more reduced products. The placement and connections among acids in the typical central metabolic network are presented along with the use of a number of specific non-native enzymes to enhance routes to high production, where available alternative pathways and strategies are discussed. While many organic acids are derived from a few precursors within central metabolism, each organic acid has its own special requirements for high production and best compatibility with host physiology.


Mandibular anatomy related to sagittal split ramus osteotomy in Koreans.
Sagittal split ramus osteotomy (SSRO) is one of the surgical techniques used to correct mandibular deformities. In order to prevent many surgical anatomical problems, we observed the anatomical structures related to SSRO. In dry mandibles of Koreans, lingular tips were located somewhat posteriorly and superiorly on the mandibular ramus. On the coronal sections of mandible, the mean cortical width of facial cortex was increased toward the ramus region while the lingual cortex was thinnest in the ramus region. On the same sections, all the fusion points of the buccal and lingual cortical plate were located above the mandibular lingula and beneath the mandibular notch. So, performing the SSRO on Koreans, medial horizontal osteotomy should be done through the superior aspect of the mandibular lingula. The cut line is extended 5-8 mm posterior to the mandibular lingula to preserve sufficient cortical width to strengthen the involved osseous segments and reduce possible surgical complications.


Receptors for prostaglandin E(2) that regulate cellular immune responses in the mouse.
Production of prostaglandin E(2) (PGE(2)) is enhanced during inflammation, and this lipid mediator can dramatically modulate immune responses. There are four receptors for PGE(2) (EP1-EP4) with unique patterns of expression and different coupling to intracellular signaling pathways. To identify the EP receptors that regulate cellular immune responses, we used mouse lines in which the genes encoding each of the four EP receptors were disrupted by gene targeting. Using the mixed lymphocyte response (MLR) as a model cellular immune response, we confirmed that PGE(2) has potent antiproliferative effects on wild-type responder cells. The absence of either the EP1 or EP3 receptors did not alter the inhibitory response to PGE(2) in the MLR. In contrast, when responder cells lacked the EP2 receptor, PGE(2) had little effect on proliferation. Modest resistance to PGE(2) was also observed in EP4-/- responder cells. Reconstitution experiments suggest that EP2 receptors primarily inhibit the MLR through direct actions on T cells. Furthermore, PGE(2) modulates macrophage function by activating the EP4 receptor and thereby inhibiting cytokine release. Thus, PGE(2) regulates cellular immune responses through distinct EP receptors on different immune cell populations: EP2 receptors directly inhibit T cell proliferation while EP2 and EP4 receptors regulate antigen presenting cells functions.


Dose-response meta-analysis on coffee, tea and caffeine consumption with risk of Parkinson's disease.
A dose-response meta-analysis was carried out between Parkinson's disease (PD) risk, and coffee, tea and caffeine consumption. A comprehensive search was carried out to identify eligible studies. The fixed or random effect model was used based on heterogeneity test. The dose-response relationship was assessed by restricted cubic spline. A total of 13 articles involving 901 764 participants for coffee, eight articles involving 344 895 participants for tea and seven articles involving 492 724 participants for caffeine were included. A non-linear relationship was found between coffee consumption and PD risk overall, and the strength of protection reached the maximum at approximately 3 cups/day (smoking-adjusted relative risk: 0.72, 95% confidence interval 0.65-0.81). A linear relationship was found between tea and caffeine consumption, and PD risk overall, and the smoking-adjusted risk of PD decreased by 26% and 17% for every two cups/day and 200 mg/day increments, respectively. The association of coffee and tea consumption with PD risk was stronger for men than that for women, and the association of caffeine consumption with PD risk was stronger for ever users of hormones than that for never users of hormones among postmenopausal women. The aforementioned associations were weaker for USA relative to Europe or Asia. A linear dose-relationship for decreased PD risk with tea and caffeine consumption was found, whereas the strength of protection reached a maximum at approximately 3 cups/day for coffee consumption overall. Further studies are required to confirm the findings.


Structural Basis of Tail-Anchored Membrane Protein Biogenesis by the GET Insertase Complex.
Membrane protein biogenesis faces the challenge of chaperoning hydrophobic transmembrane helices for faithful membrane insertion. The guided entry of tail-anchored proteins (GET) pathway targets and inserts tail-anchored (TA) proteins into the endoplasmic reticulum (ER) membrane with an insertase (yeast Get1/Get2 or mammalian WRB/CAML) that captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively). Here, we present cryo-electron microscopy reconstructions, native mass spectrometry, and structure-based mutagenesis of human WRB/CAML/TRC40 and yeast Get1/Get2/Get3 complexes. Get3 binding to the membrane insertase supports heterotetramer formation, and phosphatidylinositol binding at the heterotetramer interface stabilizes the insertase for efficient TA insertion in vivo. We identify a Get2/CAML cytoplasmic helix that forms a "gating" interaction with Get3/TRC40 important for TA insertion. Structural homology with YidC and the ER membrane protein complex (EMC) implicates an evolutionarily conserved insertion mechanism for divergent substrates utilizing a hydrophilic groove. Thus, we provide a detailed structural and mechanistic framework to understand TA membrane insertion.


Maternal antenatal body mass index gains as predictors of large-for-gestational-age infants and cesarean deliveries in Japanese singleton pregnancies.
Greater antenatal weight or body mass index (BMI) gains may lead to larger fetuses and thus increase the risk for operative deliveries, such as cesarean deliveries. In order to examine the effect of weekly maternal weight and BMI changes on large-for-gestational-age (LGA) infants and cesarean delivery, delivery records from overall healthy women were analyzed. Singleton, term delivery records from January to December 2003, at three obstetric departments (urban, suburban, rural) in Japan (1617 records) were analyzed. Multivariate logistic regression analyses were applied in order to estimate the effect of maternal pre-pregnancy body size, higher maternal weight or BMI gains by calculating the odds ratios for LGA infants and cesarean deliveries. Maternal pre-pregnancy overweight, primiparity, and BMI gains of more than 0.13 per week were independent factors positively related to LGA, and maternal underweight was negatively related. Urban hospital setting, maternal age 35 years or older, primiparity, and BMI gains of more than 0.13 per week independently increased the odds ratios for cesarean delivery. Weight or BMI gains between two-week intervals (26-28 weeks, 32-34 weeks) were not useful for predicting either LGA or cesarean delivery. LGA was unrelated to cesarean delivery risk. Applying BMI gains per week enables target weight gains to be set according to the mother's height, which may be useful in reducing risks for LGA and cesarean delivery, especially in shorter women. Further investigation may be needed to explore the practicality and effectiveness of advising women to gain weight according to their body height.


Prolactin secretion and corpus luteum function in women with luteal phase deficiency.
Luteal phase deficiency (LPD) as a clinical infertility problem is considered to have a heterogeneous etiology. Hyperprolactinemia has long been considered a causative factor of LPD. In this context we investigated PRL secretion in 18 women with LPD. All of the subjects were infertile with 2 out of phase (greater than 2 days) endometrial biopsies; 10 of the women also had daily blood samples, this latter subgroup had significantly decreased integrated luteal phase progesterone (P) levels compared to normal women with in-phase biopsies. PRL secretion was investigated as follows: 1) daily blood levels; 2) pulsatile secretion patterns in 3 cycle phase [early follicular (12 h); late follicular (12 h); midluteal (24 h)], 3) LH-PRL coupling, and 4) nocturnal patterns. Results were compared to findings in 36 normal women. The mean daily levels of PRL over the menstrual cycle were not different between the two groups (LPD, 12.1 +/- 1.5; normal, 13.8 +/- 0.8 microgram/L; P = 0.3). There was no correlation between luteal phase integrated P and PRL levels for either group. There was a small difference in the PRL pulse amplitude in the early follicular phase between the LPD and normal women (2.6 +/- 0.3 vs. 5.5 +/- 1.3 micrograms/L; P less than 0.05). There were no significant differences between groups in PRL pulse frequency or mean level during the 12 or 24 h in any cycle phase. There was an equivalent amount of LH-PRL pulse coupling in both groups in all three cycle phases. Diurnal and nocturnal PRL secretion was studied by breaking the 24 h data (midluteal) into day (0700-2300 h) and night (2300-0700) segments. Mean PRL levels were higher at night in both groups (LPD, 15.9 vs. 12.6; normal, 15.4 vs. 9.3 micrograms/L; P less than 0.05), as expected. There were no differences in nocturnal PRL secretory patterns between the two groups. In summary, we have serious reservations whether abnormalities in PRL secretion are a common or integral part of the pathophysiology of LPD. From previous work we know these subtle abnormalities in PRL secretion in LPD are associated with definite abnormalities in gonadotropin secretion. We believe these gonadotropin abnormalities are probably more significant in terms of decreased P secretion.


Ocular lesions associated with Trypanosoma evansi in experimentally infected goats.
Ocular lesions associated with Trypanosoma spp. infection have been described in man and many animal species. However, loss of vision has not been demonstrated in humans presenting Chagas disease or in animals affected by different trypanosome species. In order to assess the possible ocular disorders caused by Trypanosoma evansi infection, six goats were inoculated with 1 x 10(5) T. evansi and maintained for 12 months and four goats were used as control. The inoculated animals became positive at serological and parasitological tests at 1-month post-inoculation and showed a subclinical course of the disease. Unilateral superficial corneal ulceration and retinochoroiditis were observed in two inoculated animals. Data from ocular neurologic examination and electroretinography showed no significant differences between inoculated and non-inoculated goats. It could be concluded that Trypanosoma evansi can produce ocular lesion but without apparent loss of vision in goats.


Gel-based and gel-free proteomic analysis of Nicotiana tabacum trichomes identifies proteins involved in secondary metabolism and in the (a)biotic stress response.
Nicotiana tabacum leaves are covered by trichomes involved in the secretion of large amounts of secondary metabolites, some of which play a major role in plant defense. However, little is known about the metabolic pathways that operate in these structures. We undertook a proteomic analysis of N. tabacum trichomes in order to identify their protein complement. Efficient trichome isolation was obtained by abrading frozen leaves. After homogenization, soluble proteins and a microsomal fraction were prepared by centrifugation. Gel-based and gel-free proteomic analyses were then performed. 2-DE analysis of soluble proteins led to the identification of 1373 protein spots, which were digested and analyzed by MS/MS, leading to 680 unique identifications. Both soluble proteins and microsomal fraction were analyzed by LC MALDI-MS/MS after trypsin digestion, leading to 858 identifications, many of which had not been identified after 2-DE, indicating that the two methods complement each other. Many enzymes putatively involved in secondary metabolism were identified, including enzymes involved in the synthesis of terpenoid precursors and in acyl sugar production. Several transporters were also identified, some of which might be involved in secondary metabolite transport. Various (a)biotic stress response proteins were also detected, supporting the role of trichomes in plant defense.


Vitamin D-dependent induction of cathelicidin in human macrophages results in cytotoxicity against high-grade B cell lymphoma.
Infiltration by macrophages represents a characteristic morphological hallmark in high-grade lymphatic malignancies such as Burkitt's lymphoma (BL). Although macrophages can, in principle, target neoplastic cells and mediate antibody-dependent cellular cytotoxicity (ADCC), tumor-associated macrophages (TAMs) regularly fail to exert direct cytotoxic functions. The underlying mechanisms responsible for this observation remain unclear. We demonstrate that inflammatory M1 macrophages kill proliferating high-grade B cell lymphoma cells by releasing the antimicrobial peptide cathelicidin in a vitamin D-dependent fashion. We show that cathelicidin directly induces cell death by targeting mitochondria of BL cells. In contrast, anti-inflammatory M2 macrophages and M2-like TAMs in BL exhibit an altered vitamin D metabolism, resulting in a reduced production of cathelicidin and consequently in inability to lyse BL cells. However, treatment of M2 macrophages with the bioactive form of vitamin D, 1,25D3, or a vitamin D receptor agonist effectively induces cathelicidin production and triggers tumoricidal activity against BL cells. Furthermore, rituximab-mediated cytotoxicity of vitamin D-treated M2 macrophages is cathelicidin-dependent. Finally, vitamin D treatment of 25-hydroxyvitamin D (25D)-deficient volunteers in vivo or primary TAMs in vitro improves rituximab-mediated ADCC against B cell lymphoma cells. These data indicate that activation of the vitamin D signaling pathway activates antitumor activity of TAMs and improves the efficacy of ADCC.


Evolutionary dynamics and genetic diversity from three genes of Anguillid rhabdovirus.
Wild freshwater eel populations have dramatically declined in recent past decades in Europe and America, partially through the impact of several factors including the wide spread of infectious diseases. The anguillid rhabdoviruses eel virus European X (EVEX) and eel virus American (EVA) potentially play a role in this decline, even if their real contribution is still unclear. In this study, we investigate the evolutionary dynamics and genetic diversity of anguiillid rhabdoviruses by analysing sequences from the glycoprotein, nucleoprotein and phosphoprotein (P) genes of 57 viral strains collected from seven countries over 40 years using maximum-likelihood and Bayesian approaches. Phylogenetic trees from the three genes are congruent and allow two monophyletic groups, European and American, to be clearly distinguished. Results of nucleotide substitution rates per site per year indicate that the P gene is expected to evolve most rapidly. The nucleotide diversity observed is low (2-3 %) for the three genes, with a significantly higher variability within the P gene, which encodes multiple proteins from a single genomic RNA sequence, particularly a small C protein. This putative C protein is a potential molecular marker suitable for characterization of distinct genotypes within anguillid rhabdoviruses. This study provides, to our knowledge, the first molecular characterization of EVA, brings new insights to the evolutionary dynamics of two genotypes of Anguillid rhabdovirus, and is a baseline for further investigations on the tracking of its spread.


Demographic and socioeconomic barriers and treatment seeking behaviors of patients with infectious keratitis requiring therapeutic penetrating keratoplasty.
To understand demographic and socioeconomic barriers and treatment-seeking behaviors of patients with infectious keratitis requiring therapeutic penetrating keratoplasty (TPK) in a developing country. This prospective non-comparative questionnaire- based study included all patients presenting to Aravind Eye Hospital, Madurai with infectious keratitis that eventuated to TPK between November 2015 and October 2016. A structured questionnaire was administered on post-operative day 3 to collect data on the demographic details, predisposing factors, prior treatment received, and treatment expenditures. In total, 227 patients underwent TPK between November 2015 and October 2016 for infectious keratitis. The majority of patients were males (n = 132, 58.1%), illiterate (n = 129, 56.8%), and had a family monthly income of less than INR 6000 (n = 142, 62.5%). Most of the patients (n = 163, 71.8%) had prior treatment with an ophthalmologist before presenting to our hospital. The mean distance travelled to reach our centre was 269.2 ± 298.5 km. The mean duration of disease before the presentation was 20.3 ± 21.1 days. Corneal smear was positive for fungus in 163 (88.1%) and Aspergillus was the most commonly isolated fungi in 55 (41.3%) cultures. The mean total cost of treatment was INR 8752.87 ± 7615.39 per patient. There was a positive correlation between the duration of the disease (rho 0.19, P = 0.0034) and the costs of treatment (rho 0.2, P = 0.0024) with the distance travelled by the patient. Patients who travelled a farther distance had a delayed onset of presentation and spent significantly more than their respective counterparts.


Dispositional mindfulness moderates the effects of stress among adolescents: rumination as a mediator.
Recent research has demonstrated that higher levels of mindfulness are associated with greater psychological and physical health. However, the majority of this research has been conducted with adults; research is only beginning to examine the effects of mindfulness among adolescents. Further, research into adolescent mindfulness has typically conceptualized mindfulness as a unidimensional phenomenon and has not yet examined multidimensional models of mindfulness that have emerged in the adult literature. Further, the mechanisms through which mindfulness influences these outcomes are presently unclear. The present study examined the effects of three facets of mindfulness among adolescents. Seventy-eight adolescents (61% female, 94% Caucasian, M age = 16) completed a measure of dispositional mindfulness at baseline. Participants then completed measures of daily stress, dysphoric affect, and state rumination over a 7-day period. Multilevel modeling analyses revealed that facets of mindfulness (i.e., nonreactivity and nonjudgment) were associated with lower levels of dysphoric mood. Mindfulness interacted with daily stress to predict later dysphoria; less mindful individuals were particularly vulnerable to the negative effects of stress. Finally, analyses demonstrated that the effect of the Mindfulness × Stress Moderation was significantly mediated by increases in daily rumination. These findings support the importance of mindfulness among adolescents and help to elucidate the mechanisms through which mindfulness influences psychological health.


Functional insights into the role of nuclear-retained long noncoding RNAs in gene expression control in mammalian cells.
The mammalian genome harbors thousands of long noncoding RNA (lncRNA) genes. Recent studies have indicated the involvement of several of these lncRNAs in the regulation of gene expression. lncRNAs play crucial roles in various biological processes ranging from epigenetic gene regulation, transcriptional control,to post-transcriptional regulation. lncRNAs are localized in various subcellular compartments, and major proportion of these are retained in the cell nucleus and could be broadly classified as nuclear-retained lncRNAs (nrRNAs). Based on the identified functions,members of the nrRNAs execute diverse roles, including providing architectural support to the hierarchical subnuclear organization and influencing the recruitment of chromatin modifier factors to specific chromatin sites. In this review, we will summarize the recently described roles of mammalian nrRNAs in controlling gene expression by influencing chromatin organization, transcription,pre-mRNA processing, nuclear organization, and their involvement in disease.


Visual-somatosensory integration in aging: does stimulus location really matter?
Individuals are constantly bombarded by sensory stimuli across multiple modalities that must be integrated efficiently. Multisensory integration (MSI) is said to be governed by stimulus properties including space, time, and magnitude. While there is a paucity of research detailing MSI in aging, we have demonstrated that older adults reveal the greatest reaction time (RT) benefit when presented with simultaneous visual-somatosensory (VS) stimuli. To our knowledge, the differential RT benefit of visual and somatosensory stimuli presented within and across spatial hemifields has not been investigated in aging. Eighteen older adults (Mean = 74 years; 11 female), who were determined to be non-demented and without medical or psychiatric conditions that may affect their performance, participated in this study. Participants received eight randomly presented stimulus conditions (four unisensory and four multisensory) and were instructed to make speeded foot-pedal responses as soon as they detected any stimulation, regardless of stimulus type and location of unisensory inputs. Results from a linear mixed effect model, adjusted for speed of processing and other covariates, revealed that RTs to all multisensory pairings were significantly faster than those elicited to averaged constituent unisensory conditions (p < 0.01). Similarly, race model violation did not differ based on unisensory spatial location (p = 0.41). In summary, older adults demonstrate significant VS multisensory RT effects to stimuli both within and across spatial hemifields.


Mesenchymal Stem Cell Treatment for Steroid Refractory Graft-versus-Host Disease in Children: A Pilot and First Study from Turkey.
This study evaluated the efficacy of mesenchymal stem cells (MSCs) from bone marrow of a third-party donor for refractory aGVHD. We report the first experience using MSCs to treat refractory aGVHD in 33 pediatric patients undergoing allogeneic HSCT from Turkey. Totally, 68 doses of bone marrow derived MSCs were infused. The median dose of MSC was 1.18 × 10(6) cells per kg body weight. Overall, complete response (CR) was documented in 18 patients, partial response (PR) was documented in 7 patients, and no response (NR) was documented in 8 patients. The 2-year estimated probability of overall survival (OS) for patients achieving CR and PR/NR was 63.8% and 29.4%, respectively (p = 0.0002). While the cumulative incidence of transplant related mortality (TRM) at day 100 after first MSC infusion was 46.6% in PR/NR patients, there was no any TRM at day 100 after first MSC infusion in CR patients (p = 0.001). Twelve patients developed chronic GVHD (cGVHD); eight of them were alive, with five having extensive disease and three having limited disease. In conclusion, MSCs appear to be safe and effective treatment option for pediatric patients with steroid refractory aGVHD. But the efficacy of MSCs on cGVHD in aGVHD patients treated with MSCs seems to be limited.


Structural Diversity and Bioactivities of Peptaibol Compounds From the Longibrachiatum Clade of the Filamentous Fungal Genus Trichoderma.
This study examined the structural diversity and bioactivity of peptaibol compounds produced by species from the phylogenetically separated Longibrachiatum Clade of the filamentous fungal genus Trichoderma, which contains several biotechnologically, agriculturally and clinically important species. HPLC-ESI-MS investigations of crude extracts from 17 species of the Longibrachiatum Clade (T. aethiopicum, T. andinense, T. capillare, T. citrinoviride, T. effusum, T. flagellatum, T. ghanense, T. konilangbra, T. longibrachiatum, T. novae-zelandiae, T. pinnatum, T. parareesei, T. pseudokoningii, T. reesei, T. saturnisporum, T. sinensis, and T. orientale) revealed several new and recurrent 20-residue peptaibols related to trichobrachins, paracelsins, suzukacillins, saturnisporins, trichoaureocins, trichocellins, longibrachins, hyporientalins, trichokonins, trilongins, metanicins, trichosporins, gliodeliquescins, alamethicins and hypophellins, as well as eight 19-residue sequences from a new subfamily of peptaibols named brevicelsins. Non-ribosomal peptide synthetase genes were mined from the available genome sequences of the Longibrachiatum Clade. Their annotation and product prediction were performed in silico and revealed full agreement in 11 out of 20 positions regarding the amino acids predicted based on the signature sequences and the detected amino acids incorporated. Molecular dynamics simulations were performed for structural characterization of four selected peptaibol sequences: paracelsins B, H and their 19-residue counterparts brevicelsins I and IV. Loss of position R6 in brevicelsins resulted in smaller helical structures with higher atomic fluctuation for every residue than the structures formed by paracelsins. We observed the formation of highly bent, almost hairpin-like, helical structures throughout the trajectory, along with linear conformation. Bioactivity tests were performed on the purified peptaibol extract of T. reesei on clinically and phytopathologically important filamentous fungi, mammalian cells, and Arabidopsis thaliana seedlings. Porcine kidney cells and boar spermatozoa proved to be sensitive to the purified peptaibol extract. Peptaibol concentrations ≥0.3 mg ml-1 deterred the growth of A. thaliana. However, negative effects to plants were not detected at concentrations below 0.1 mg ml-1, which could still inhibit plant pathogenic filamentous fungi, suggesting that those peptaibols reported here may have applications for plant protection.


Dynamic interactions between visual working memory and saccade target selection.
Recent psychophysical experiments have shown that working memory for visual surface features interacts with saccadic motor planning, even in tasks where the saccade target is unambiguously specified by spatial cues. Specifically, a match between a memorized color and the color of either the designated target or a distractor stimulus influences saccade target selection, saccade amplitudes, and latencies in a systematic fashion. To elucidate these effects, we present a dynamic neural field model in combination with new experimental data. The model captures the neural processes underlying visual perception, working memory, and saccade planning relevant to the psychophysical experiment. It consists of a low-level visual sensory representation that interacts with two separate pathways: a spatial pathway implementing spatial attention and saccade generation, and a surface feature pathway implementing color working memory and feature attention. Due to bidirectional coupling between visual working memory and feature attention in the model, the working memory content can indirectly exert an effect on perceptual processing in the low-level sensory representation. This in turn biases saccadic movement planning in the spatial pathway, allowing the model to quantitatively reproduce the observed interaction effects. The continuous coupling between representations in the model also implies that modulation should be bidirectional, and model simulations provide specific predictions for complementary effects of saccade target selection on visual working memory. These predictions were empirically confirmed in a new experiment: Memory for a sample color was biased toward the color of a task-irrelevant saccade target object, demonstrating the bidirectional coupling between visual working memory and perceptual processing.


Hedgehog signaling is critical for maintenance of the adult coronary vasculature in mice.
Hedgehog (HH) signaling has emerged as a critical pathway involved in the pathogenesis of a variety of tumors. As a result, HH antagonists are currently being evaluated as potential anticancer therapeutics. Conversely, activation of HH signaling in the adult heart may be beneficial, as HH agonists have been shown to increase coronary vessel density and improve coronary function after myocardial infarction. To investigate a potential homeostatic role for HH signaling in the adult heart, we ablated endogenous HH signaling in murine myocardial and perivascular smooth muscle cells. HH signaling was required for proangiogenic gene expression and maintenance of the adult coronary vasculature in mice. In the absence of HH signaling, loss of coronary blood vessels led to tissue hypoxia, cardiomyocyte cell death, heart failure, and subsequent lethality. We further showed that HH signaling specifically controlled the survival of small coronary arteries and capillaries. Together, these data demonstrate that HH signaling is essential for cardiac function at the level of the coronary vasculature and caution against the use of HH antagonists in patients with prior or ongoing heart disease.


Characterization of Reference Materials for Genetic Testing of CYP2D6 Alleles: A GeT-RM Collaborative Project.
Pharmacogenetic testing increasingly is available from clinical and research laboratories. However, only a limited number of quality control and other reference materials currently are available for the complex rearrangements and rare variants that occur in the CYP2D6 gene. To address this need, the Division of Laboratory Systems, CDC-based Genetic Testing Reference Material Coordination Program, in collaboration with members of the pharmacogenetic testing and research communities and the Coriell Cell Repositories (Camden, NJ), has characterized 179 DNA samples derived from Coriell cell lines. Testing included the recharacterization of 137 genomic DNAs that were genotyped in previous Genetic Testing Reference Material Coordination Program studies and 42 additional samples that had not been characterized previously. DNA samples were distributed to volunteer testing laboratories for genotyping using a variety of commercially available and laboratory-developed tests. These publicly available samples will support the quality-assurance and quality-control programs of clinical laboratories performing CYP2D6 testing.


A DNA hypermethylation module for the stem/progenitor cell signature of cancer.
Many DNA-hypermethylated cancer genes are occupied by the Polycomb (PcG) repressor complex in embryonic stem cells (ESCs). Their prevalence in the full spectrum of cancers, the exact context of chromatin involved, and their status in adult cell renewal systems are unknown. Using a genome-wide analysis, we demonstrate that ~75% of hypermethylated genes are marked by PcG in the context of bivalent chromatin in both ESCs and adult stem/progenitor cells. A large number of these genes are key developmental regulators, and a subset, which we call the "DNA hypermethylation module," comprises a portion of the PcG target genes that are down-regulated in cancer. Genes with bivalent chromatin have a low, poised gene transcription state that has been shown to maintain stemness and self-renewal in normal stem cells. However, when DNA-hypermethylated in tumors, we find that these genes are further repressed. We also show that the methylation status of these genes can cluster important subtypes of colon and breast cancers. By evaluating the subsets of genes that are methylated in different cancers with consideration of their chromatin status in ESCs, we provide evidence that DNA hypermethylation preferentially targets the subset of PcG genes that are developmental regulators, and this may contribute to the stem-like state of cancer. Additionally, the capacity for global methylation profiling to cluster tumors by phenotype may have important implications for further refining tumor behavior patterns that may ultimately aid therapeutic interventions.


Risk of spread in adult-onset isolated focal dystonia: a prospective international cohort study.
Isolated focal dystonia can spread to muscles beyond the initially affected body region, but risk of spread has not been evaluated in a prospective manner. Furthermore, body regions at risk for spread and the clinical factors associated with spread risk are not well characterised. We sought here to prospectively characterise risk of spread in recently diagnosed adult-onset isolated focal dystonia patients. Patients enrolled in the Dystonia Coalition with isolated dystonia affecting only the neck, upper face, hand or larynx at onset of symptoms were included. Timing of follow-up visits was based on a sliding scale depending on symptom onset and ranged from 1 to 4 years. Descriptive statistics, Kaplan-Meier survival curves and Cox proportional hazard regression models were used to assess clinical characteristics associated with dystonia spread. 487 enrolled participants (68.3% women; mean age: 55.6±12.2 years) met our inclusion/exclusion criteria. Spread was observed in 50% of blepharospasm, 8% of cervical dystonia, 17% of hand dystonia and 16% of laryngeal dystonia cases. Most common regions for first spread were the oromandibular region (42.2%) and neck (22.4%) for blepharospasm, hand (3.5%) for cervical dystonia and neck for hand (12.8%) and laryngeal (15.8%) dystonia. Increased spread risk was associated with a positive family history (HR=2.18, p=0.012) and self-reported alcohol responsiveness (HR=2.59, p=0.009). Initial body region affected in isolated focal dystonia has differential risk and patterns of spread. Genetic factors likely influence the risk of spread. These findings can aid clinical prognostication and inform future investigations into potential disease-modifying treatments.