Loss of Magel2, a Candidate Gene for Features of Prader-Willi Syndrome, Impairs Reproductive Function in Mice

Background: MAGEL2 is one of several genes typically inactivated in the developmental obesity disorder Prader-Willi syndrome (PWS). The physiological consequences of loss of MAGEL2, but without the concurrent loss of other PWS genes, are not well understood. Gene-targeted mutation of Magel2 in mice disrupts circadian rhythm and metabolism causing reduced total activity, reduced weight gain before weaning, and increased adiposity after weaning. Principal Findings: We now show that loss of Magel2 in mice causes reduced fertility in both males and females through extended breeding intervals and early reproductive decline and termination. Female Magel2-null mice display extended and irregular estrous cycles, while males show decreased testosterone levels, and reduced olfactory preference for female odors. Conclusions: Our results suggest that loss of MAGEL2 contributes to the reproductive deficits seen in people with PWS, and further highlights the role of normal circadian rhythm in the maintenance of fertility

Morgellons disease, illuminating an undefined illness: a case series

<p>Abstract</p> <p>Introduction</p> <p>This review of 25 consecutive patients with Morgellons disease (MD) was undertaken for two primary and extremely fundamental reasons. For semantic accuracy, there is only one "proven" MD patient: the child first given that label. The remainder of inclusive individuals adopted the label based on related descriptions from 1544 through 1884, an internet description quoted from Sir Thomas Browne (1674), or was given the label by practitioners using similar sources. Until now, there has been no formal characterization of MD from detailed examination of all body systems. Our second purpose was to differentiate MD from Delusions of Parasitosis (DP), another "informal" label that fit most of our MD patients. How we defined and how we treated these patients depended literally on factual data that would determine outcome. How they were labeled in one sense was irrelevant, except for the confusing conflict rampant in the medical community, possibly significantly skewing treatment outcomes.</p> <p>Case presentation</p> <p>Clinical information was collected from 25 of 30 consecutive self-defined patients with Morgellons disease consisting of laboratory data, medical history and physical examination findings. Abnormalities were quantified and grouped by system, then compared and summarized, but the numbers were too small for more complex mathematical analysis. The quantification of physical and laboratory abnormalities allowed at least the creation of a practical clinical boundary, separating probable Morgellon<it>s</it> from non-Morgellons patients. All the 25 patients studied meet the most commonly used DP definitions.</p> <p>Conclusions</p> <p>These data suggest Morgellons disease can be characterized as a physical human illness with an often-related delusional component in adults. All medical histories support that behavioral aberrancies onset only after physical symptoms. The identified abnormalities include both immune deficiency and chronic inflammatory markers that correlate strongly with immune cytokine excess. The review of 251 current NLM DP references leads us to the possibility that Morgellons disease and DP are grossly truncated labels of the same illness but with the reversal of the cause-effect order. Further, the patients' data suggest that both illnesses have an infectious origin.</p

The vaccinia chondroitin sulfate binding protein drives host membrane curvature to facilitate fusion

Cellular attachment of viruses determines their cell tropism and species specificity. For entry, vaccinia, the prototypic poxvirus, relies on four binding proteins and an eleven-protein entry fusion complex. The contribution of the individual virus binding proteins to virion binding orientation and membrane fusion is unclear. Here, we show that virus binding proteins guide side-on virion binding and promote curvature of the host membrane towards the virus fusion machinery to facilitate fusion. Using a membrane-bleb model system together with super-resolution and electron microscopy we find that side-bound vaccinia virions induce membrane invagination in the presence of low pH. Repression or deletion of individual binding proteins reveals that three of four contribute to binding orientation, amongst which the chondroitin sulfate binding protein, D8, is required for host membrane bending. Consistent with low-pH dependent macropinocytic entry of vaccinia, loss of D8 prevents virion-associated macropinosome membrane bending, disrupts fusion pore formation and infection. Our results show that viral binding proteins are active participants in successful virus membrane fusion and illustrate the importance of virus protein architecture for successful infection.</p

Developing a framework to incorporate real-world evidence in cancer drug funding decisions : The Canadian Real-world Evidence for Value of Cancer Drugs (CanREValue) collaboration

Background Oncology therapy is becoming increasingly more expensive and challenging the affordability and sustainability of drug programmes around the world. When new drugs are evaluated, health technology assessment organisations rely on clinical trials to inform funding decisions. However, clinical trials are not able to assess overall survival and generalises evidence in a real-world setting. As a result, policy makers have little information on whether drug funding decisions based on clinical trials ultimately yield the outcomes and value for money that might be expected. Objective The Canadian Real-world Evidence for Value of Cancer Drugs (CanREValue) collaboration, consisting of researchers, recommendation-makers, decision makers, payers, patients and caregivers, are developing and testing a framework for Canadian provinces to generate and use real-world evidence (RWE) for cancer drug funding in a consistent and integrated manner. Strategy The CanREValue collaboration has established five formal working groups (WGs) to focus on specific processes in the generation and use of RWE for cancer drug funding decisions in Canada. The different RWE WGs are: (1) Planning and Drug Selection; (2) Methods; (3) Data; (4) Reassessment and Uptake; (5) Engagement. These WGs are acting collaboratively to develop a framework for RWE evaluation, validate the framework through the multiprovince RWE projects and help to integrate the final RWE framework into the Canadian healthcare system. Outcomes The framework will enable the reassessment of cancer drugs, refinement of funding recommendations and use of novel funding mechanisms by decision-makers/payers across Canada to ensure the healthcare system is providing clinical benefits and value for money

Regionally reduced brain volume, altered serotonin neurochemistry, and abnormal behavior in mice null for the circadian rhythm output gene Magel2.

Magel2 belongs to the MAGE/necdin family of proteins, which have roles in cell cycle, differentiation, and apoptosis. The Magel2 gene is expressed in various brain regions, most notably the hypothalamus. Mice with a targeted deletion of Magel2 display hypoactivity, blunted circadian rhythm, decreased fertility, and increased adiposity. The human ortholog, MAGEL2, is one of a set of paternally expressed, imprinted genes inactivated in most cases of Prader-Willi syndrome, a complex neurodevelopmental disorder. To explore the role of Magel2, brain morphology, brain neurochemistry, and behavior were measured in Magel2-null mice. Brain volume was reduced in specific regions, particularly in the parieto-temporal lobe of the cerebral cortex, the amygdala, the hippocampus, and the nucleus accumbens, as measured by quantitative magnetic resonance imaging. Abnormal neurochemistry was detected in brain samples from adult mice, consisting of decreased serotonin and 5-hydroxyindoleacetic acid in the cortex and the hypothalamus, and decreased dopamine in the hypothalamus. Magel2-null mice displayed relatively normal motor and learning abilities, but exhibited abnormal behavior in novel environments. This study lends support to the important role of the circadian rhythm output gene Magel2 in brain structure and behavior. Ó 2009 Wiley-Liss, Inc. Key words: Prader-Willi syndrome; anxiety; magnetic resonance imaging; circadian rhythm; imprinting INTRODUCTION Magel2 is a member of the Type II melanoma-associated antigen gene (MAGE) protein family, which share a protein-protein interaction domain called the MAGE homology/conserved domain 1085 Neuropsychiatric Genetics 1995]. The MAGE homology domain of several Type II MAGE proteins (necdin, MAGE-G1, MAGED1) binds to multiple transmembrane receptors involved in intracellular signaling MAGEL2 and NDN, the gene that encodes the MAGE protein necdin, are among a small set of genes that are typically inactivated in Prader-Willi syndrome (PWS). PWS is a congenital disorder characterized by symptoms of varying severity among affected individuals: intellectual disability, hypotonia, short stature, childhood-onset hyperphagia often leading to obesity, excessive sleepiness, neuroendocrine abnormalities, and incomplete sexual development We recently described a mouse strain carrying a gene-targeted lacZ insertion into the Magel2 locus, creating a null Magel2 allele MATERIALS AND METHODS Magel2-Null Mice All animal studies were conducted in accordance with the Canadian Council on Animal Care Guidelines and Policies with approval from the local Animal Policy and Welfare Committees. Magel2-null mice carry a replacement of the Magel2 gene with a LacZ reporter cassette, resulting in complete loss of Magel2 function in heterozygotes with a paternally derived gene-targeted allele Sample Preparation for MRI Magel2-null mice (n ¼ 6) and their wild-type littermates (n ¼ 6) were anesthetized at 26 weeks of age with a combination of Ketamine (100 mg/kg, Pfizer, Kirkland, QC, Canada) and Rompun (20 mg/kg, Bayer, Inc., Toronto, ON, Canada) via intraperitoneal injection. A previously described sample preparation protocol for scanning was used with slight modifications MRI Image Acquisition A multi-channel 7.0-T MRI scanner (Varian, Inc., Palo Alto, CA) with a 6 cm inner bore diameter insert gradient set was used to acquire anatomical images of brains within skulls. Prior to imaging, the samples were removed from the contrast agent solution, blotted and placed into 13 mm diameter plastic tubes filled with a protonfree susceptibility-matching fluid 3 M Corp., St. Paul, MN). Three custom-built, 14 mm diameter solenoid coils with a length of 18.3 mm and over wound ends were used to image three brains in parallel. Parameters used in the scans were optimized for gray/white matter contrast: a T2-weighted, 3D fast spin-echo sequence, with TR/TE ¼ 325/32 msec, four averages, field-of-view 14 mm  14 mm  25 mm and matrix size ¼ 432  432  780 giving an image with 32 mm isotropic voxels. Total imaging time was 11.3 hr . Image Processing The 32 mm isotropic resolution T2-weighted MRI scans were nonlinearly aligned to a three dimensional atlas of the mouse brain with 62 structures identified The end result was that all 12 scans were deformed into exact alignment with each other in an unbiased fashion. This allowed for the analysis of the deformations needed to take each mouse&apos;s anatomy into this final atlas space, the goal being to model how the deformation fields relate to genotype. Correspondence with the 3D atlas was obtained by nonlinear alignment of the final stage average MRI with the 40-mouse average MRI upon which the atlas is based MRI Analysis Local differences in brain shape related to genotype were assessed by analysis of the deformation fields Neurochemical Analysis Brain regions were dissected from embryonic or adult Magel2-null or wild-type littermate control mice, snap frozen on dry ice, then stored at À80 C. Brain samples were processed for HPLC combined with fluorescence detection to measure levels of biogenic amines (noradrenaline (NA), dihydroxyphenylacetic acid (DOPAC), dopamine (DA), 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA), serotonin (5-HT) and amino acids (Trp, Asp, Glu, Asn, Ser, Gln, Gly, Taur, Ala, GABA)) as described Behavioral Analysis Male Magel2-null mice and their control littermates were tested in the following order at the Centre for Modeling Human Disease: modified SHIRPA for general health, appearance, and neurological reflexes The contextual and cued fear conditioning test was performed using the Video Fear Conditioning system (Med Associates, St. Albans, VT) essentially as described The custom apparatus for the beam test is made up of two platforms (25 cm  35 cm) that are connected with a 90 cm long round beam (18 mm diameter) suspended 50 cm above the floor. One platform was brightly illuminated while the opposite platform MERCER ET AL. 1087 was dark and contained a box providing an enclosed safety area for the mouse. Four consecutive training trials to assist the mouse in navigating the beam were performed on the first day. On the second day, a single trial that recorded the latency to traverse the beam and the number of times the hind feet slipped off was performed. The one-trial object recognition task Behavioral data were analyzed statistically by t-test except the horizontal and vertical activity profile, the 3-day Rotarod test, and the fear-conditioning test, which were analyzed by two-way ANOVA with Bonferroni post-tests, using GraphPad Prism. Probabilities &lt;0.05 were deemed significant. RESULTS Expression of Magel2 in the Adult Mouse Brain We previously outlined expression of Magel2 in the embryonic brain by RNA in situ hybridization General Behavior and Health of Magel2-Null Mice No overt differences in the acquisition of developmental milestones (e.g., physical development, rooting reflex, righting reflex, forelimb/hind limb grasping, or locomotor behavior Neuroanatomical Abnormalities in Magel2-Null Mice Measured by MRI No gross abnormalities in sections of the Magel2-null mouse brains were detected by Nissl staining (not shown). To examine the brain more finely and without the confounding effect of removal from the skull, very high resolution MRI combined with computer analysis was performed. MRI analysis revealed reduced brain volume in the mutant mice (3.4% smaller than control brain, P &lt; 0.01). We used a statistical map of the Jacobian determinant that illustrates the expansion and contraction of tissue based on genotype, to find regions of significant change. In order to account for an inflated amount of false positive findings due to the number of statistical tests employed, the FDR technique was applied The reduced concentration of serotonin or dopamine could be caused by a reduction in the number of dopaminergic or serotonergic neurons in the Magel2-null mouse brains. To examine this possibility, we first examined the location and number of dopaminergic neurons (immunoreactive with an antibody against TH and visualized by confocal microscopy) in the hypothalamus in adult and Magel2-null male mouse brain sections. We found a comparable number of positively staining neurons in both genotypes, and no differences in the appearance of the TH-positive neurons groups in the pre-optic region, the medial basal region, or the medial dorsal region of the hypothalamus (A11-A15 dopaminergic cell groups, Suppl. AMERICAN JOURNAL OF MEDICAL GENETICS PART B with an anti-5-HT antibody revealed no differences in the location or number of serotonergic neurons in the cell groups in the brain stem (Suppl. Magel2-Null Tests of Anxiety and Learning The elevated plus maze is a more specific test of anxiety in a novel environment. The time spent in the open or closed arms, time spent on the end of the open arms and on the central platform, time spent freezing in the open and freezing in the center, the number of passes between closed arms, the number of risk assessments, and the number of head dips were all measured, but no significant differences between genotypes were detected (data not shown). We then used a Pavlovian fear-conditioning test that measures percent of time spent with total lack of movement (freezing) after an aversive stimulus, and the learning associated with this stimulus. This test also models anticipatory anxiety, and requires a combination of amygdalar and hippocampal function. Each mouse was acclimatized in a test context, then a 30 sec auditory tone was used as the conditioned stimulus paired with an aversive unconditioned stimulus, in this case a 2 sec mild foot shock at the end of the tone. After conditioning, either the test context or the tone typically elicit a state of fear even in the absence of the foot shock, in a normal mouse. This fear is manifested as freezing, and is used to measure learning when assessed after 24 hr. The Magel2-null mice tended towards an increased amount of time spent freezing during the baseline measurement on Day 1 (1.2 AE 0.7% for control mice, 12.4 AE 5.7% for the Magel2-null mice, P &lt; 0.09, all values expressed as mean AE standard error of the mean (SEM), Magel2-Null Mice Display Altered Behavior in Novel Environments As a further test to discriminate anxiety from learning, we performed a set of tests that measure reactions to novel objects in an independent cohort of male and female mice that were na€ ıve to behavioral testing. In a one-trial object recognition task, each Latency to fall from an accelerating Rotarod during a single test is significantly increased in the Magel2-null mice; **P &lt; 0.05. E: Latency to fall from an accelerating Rotarod increases for both genotypes over three consecutive days indicating normal motor learning. Latency to fall is significantly increased in the Magel2-null mice (two-way ANOVA, main effect of genotype P &lt; 0.0006). AMERICAN JOURNAL OF MEDICAL GENETICS PART B FIG. 3. Fear conditioning test and novel object scenarios reveal abnormal behavior in Magel2-null mice. A: Magel2-null mice freeze more than wildtype (WT) control mice in the test context at baseline and after the auditory tone, but have similar freezing rates after the foot shock. On Day 2, freezing in the familiar test context was not different between genotypes, but in contrast, Magel2-null mice freeze more than control at baseline in the altered context, pre-tone. Also on Day 2, the net amount of freezing in the novel context either before (pre-tone) or after (tone) the auditory tone was not different between genotypes. Data are expressed as mean AE SEM. Two-way ANOVA detected a main effect of genotype across the six freezing measurements (P &lt; 0.0001). The fold change (x) in freezing during fear conditioning is also presented as a ratio of the means. Fold changes in the test context (tone, shock, day 2 context) are compared to baseline. Fold changes in the altered context after the tone are compared to the pre-tone measurement. Magel2-null mice show a blunted fear reaction to either the test context or to the tone. B: Female Magel2-null display increased exploratory behavior when placed in a cage with two novel objects, with decreased latency to reach 38 sec total exploration time. Data are expressed as mean AE SEM, **P &lt; 0.05. C: Control mice of both sexes and male Magel2-null mice spend more time with the novel object than with the familiar object, measured after 5 min and after 24 hr, and presented as a discrimination index (the difference between the time with the novel object and the time with familiar object divided by the total time). Magel2-null female mice show no preference for a novel object after 3 min (DI ¼ 0, **P &lt; 0.05). D: Male mice of both genotypes buried similar numbers of marbles over the three min. test interval. In contrast, female Magel2-null mice buried significantly fewer marbles than controls; **P &lt; 0.05. E: Control mice consume slightly less food during their first to days in an Accuscan chamber with powdered food, but increase food consumption to normal levels by day 3 in the chamber. In contrast, Magel2-null mice have substantially reduced food intake when initially placed in the Accuscan chamber, and never attain normal levels of food intake. MERCER ET AL. 1093 mouse was removed from the home cage and acclimatized to the test cage in which two objects (e.g., plastic 15 ml tube, 10 ml syringe) were placed. The total length of time needed to accumulate 38 sec of active object exploration was recorded. Magel2-null female mice explored the objects more actively than their control littermates, accumulating 38 sec of exploration time within 141 AE 26 sec, while the control female mice required twice as long to accumulate the total exploration time (321 AE 53 sec, P &lt; 0.02) Wild-type mice typically spend 70% of their time with the novel object and 30% with the familiar object under this paradigm Our hypothesis that Magel2-null mice tend to avoid novel objects and are anxious in novel environments is supported by observations we made during a previous feeding study. In this experiment, male mice were placed in a chamber with a powdered standard chow dispenser that measured hourly food consumption over 5 days Additional Tests of Behavior Mice normally exhibit self-grooming behavior, and pathological self-grooming or excessive grooming of cage mates has been interpreted as evidence of obsessive-compulsive tendencies in mice DISCUSSION Magel2 belongs to the MAGE family of proteins, and is most closely related to MAGED1/NRAGE and necdin. Studies in cell culture and in mice have implicated necdin in neural differentiation and cell cycle exit, and structural and function deficits in the nervous system have been identified in necdin-null mice Magel2-Null Mice Have Reduced Brain Volume and Reduced Neurotransmitter Levels in Discrete Regions of the Brain Subtle changes in regional brain volume have been described in a variety of congenital and progressive genetic disorders, most 1094 AMERICAN JOURNAL OF MEDICAL GENETICS PART B prominently in human and mouse studies of schizophrenia It is difficult to establish a cause and effect relationship between neurochemical imbalances and behavior in mice, particularly as total levels combine the intracellular and extracellular pools of the neurotransmitters. For example, many studies implicating serotonin in mood and behavior use surrogate markers, such as 5-HT or 5-HIAA levels in cerebrospinal fluid or platelets, examine the activation of serotonin receptors after pharmacological intervention, or the effects of depletion of the serotonin precursor tryptophan on behavior. Nonetheless, many studies have linked altered serotonergic and dopaminergic pathways with psychiatric disorders, most notably depression, anxiety, and self-injurious or obsessive behavior Magel2-Null Mice Are Hypoactive and React Abnormally to Novel Environments We performed assays in Magel2-null mice designed to measure anxiety-like behavior, locomotion, balance, neuromuscular function, learning, and memory. Behavioral tests in animal models can provide surrogate markers for normal or pathological human behavior. In transgenic animal studies, interacting deficits in different processes can influence performance in behavior tests Balance and strength were not impaired by the Magel2 mutation, as evidenced by normal or improved function in rearing in the open field, time to cross in the beam test, increased latency to fall from the Rotarod, and equivalent time spent struggling in the tail suspension test. One interpretation of the increased latency to fall from the Rotarod is that the Magel2-null mice have normal strength and balance, as supported by the other tests of motor function, but have increased motivation not to fall, consistent with their abnormal reaction to other novel environments. We did however observe a significant reduction in open field activity in Magel2-null mice, consistent with previous findings that used running wheels to monitor 24 hr activity of Magel2-null mice MERCER ET AL. 1095 We found no evidence for learning or long-term memory deficiencies in the 3-day Rotarod test, the 24 hr novel object preference test, or the 24 hr fear-conditioning test. There was an increase in freezing in the Magel2-null mice on day 2 of the fear conditioning test, suggesting they are not grossly impaired in amygdalar or hippocampal functions required for conditioned learning over 24 hr. The interpretation of learning in the fearconditioning test was complicated by the high baseline freezing rates in the Magel2-null mice, which led to a smaller fold increase in freezing either to the context or to the tone on Day 2. The conditioned fear test revealed a significant difference between genotypes not related to learning or memory: male Magel2-null mice have increased time spent freezing under baseline conditions, indicating increased anxiety in the test chamber. It is unlikely that reduced activity accounts for this increase in freezing during the 2 min baseline measurement, as there was only minimal difference in horizontal or vertical activity between genotypes in the first 5 min of the open field test. Rather, there was a progressive decline in activity of the Magel2-null mice over the following 25 min in the open field. In a test used as a proxy for anxiety in rodents, we found no inter-genotype difference in marble burying activity in male mice, nor was there any difference in the time male mice spent with objects in the cage. In contrast, female Magel2-null mice spent more time exploring objects placed in a cage, did not display novel object preference when placed back into the test chamber after a 5 min interval, and were less likely to bury marbles placed on the surface of the bedding. In summary, the object-based tests with the female mice suggest a combination of poor short-term memory for novel objects and decreased motivation to manipulate objects in the cage. Interestingly, sex-specific differences in behavioral responses to novel objects were also observed in a serotonin-depletion mouse model of developmental brain disorders Abnormalities of Brain and Behavior: Comparison With PWS In PWS, psychiatric symptoms often develop during childhood and can include mood instability, obsessive-compulsive disorder, autism spectrum disorder, cognitive rigidity, anxiety, and addictive behavior towards to food and other substances Although the extrapolation of murine studies to human behavior must be approached with caution, studies of genetically engineered mice have successfully recapitulated the fundamental behavioral aspects of the respective human genetic disorder in many cases Almost all individuals with PWS lack expression of multiple genes, including loss of function of MAGEL2 and necdin, and most PWS candidate genes are moderately to highly expressed in the brain . It is unclear how much contribution to the PWS phenotype is made by each of the deleted genes. Comparison with other mouse strains carrying individual PWS gene deletions may be informative, although few behavioral studies have been performed to date. We previously showed that mice lacking necdin are underweight at birth A report of a child with atypical PWS carrying a chromosome deletion of 175 kb AMERICAN JOURNAL OF MEDICAL GENETICS PART B RNAs (HBII-85) (e.g., ACKNOWLEDGMENT

Identifying the science and technology dimensions of emerging public policy issues through horizon scanning

Public policy requires public support, which in turn implies a need to enable the public not just to understand policy but also to be engaged in its development. Where complex science and technology issues are involved in policy making, this takes time, so it is important to identify emerging issues of this type and prepare engagement plans. In our horizon scanning exercise, we used a modified Delphi technique [1]. A wide group of people with interests in the science and policy interface (drawn from policy makers, policy adviser, practitioners, the private sector and academics) elicited a long list of emergent policy issues in which science and technology would feature strongly and which would also necessitate public engagement as policies are developed. This was then refined to a short list of top priorities for policy makers. Thirty issues were identified within broad areas of business and technology; energy and environment; government, politics and education; health, healthcare, population and aging; information, communication, infrastructure and transport; and public safety and national security.Public policy requires public support, which in turn implies a need to enable the public not just to understand policy but also to be engaged in its development. Where complex science and technology issues are involved in policy making, this takes time, so it is important to identify emerging issues of this type and prepare engagement plans. In our horizon scanning exercise, we used a modified Delphi technique [1]. A wide group of people with interests in the science and policy interface (drawn from policy makers, policy adviser, practitioners, the private sector and academics) elicited a long list of emergent policy issues in which science and technology would feature strongly and which would also necessitate public engagement as policies are developed. This was then refined to a short list of top priorities for policy makers. Thirty issues were identified within broad areas of business and technology; energy and environment; government, politics and education; health, healthcare, population and aging; information, communication, infrastructure and transport; and public safety and national security

Large-scale differences in microbial biodiversity discovery between 16S amplicon and shotgun sequencing

Modern metagenomic environmental DNA studies are almost completely reliant on next-generation sequencing, making evaluations of these methods critical. We compare two next-generation sequencing techniques – amplicon and shotgun – on water samples across four of Brazil’s major river floodplain systems (Amazon, Araguaia, Paraná, and Pantanal). Less than 50% of phyla identified via amplicon sequencing were recovered from shotgun sequencing, clearly challenging the dogma that mid-depth shotgun recovers more diversity than amplicon-based approaches. Amplicon sequencing also revealed ~27% more families. Overall the amplicon data were more robust across both biodiversity and community ecology analyses at different taxonomic scales. Our work doubles the sampling size in similar environmental studies, and novelly integrates environmental data (e.g., pH, temperature, nutrients) from each site, revealing divergent correlations depending on which data are used. While myriad variants on NGS techniques and bioinformatic pipelines are available, our results point to core differences that have not been highlighted in any studies to date. Given the low number of taxa identified when coupling shotgun data with clade-based taxonomic algorithms, previous studies that quantified biodiversity using such bioinformatic tools should be viewed cautiously or re-analyzed. Nonetheless, shotgun has complementary advantages that should be weighed when designing projects

Translational pharmacology of an inhaled small molecule αvβ6 integrin inhibitor for idiopathic pulmonary fibrosis

The αvβ6 integrin plays a key role in the activation of transforming growth factor-β (TGFβ), a pro-fibrotic mediator that is pivotal to the development of idiopathic pulmonary fibrosis (IPF). We identified a selective small molecule αvβ6 RGD-mimetic, GSK3008348, and profiled it in a range of disease relevant pre-clinical systems. To understand the relationship between target engagement and inhibition of fibrosis, we measured pharmacodynamic and diseaserelated end points. Here we report, GSK3008348 binds to αvβ6 with high affinity in human IPF lung and reduces downstream pro-fibrotic TGFβ signaling to normal levels. In human lung epithelial cells, GSK3008348 induces rapid internalization and lysosomal degradation of the αvβ6 integrin. In the murine bleomycin-induced lung fibrosis model, GSK3008348 engages αvβ6, induces prolonged inhibition of TGFβ signaling and reduces lung collagen deposition and serum C3M, a marker of IPF disease progression. These studies highlight the potential of inhaled GSK3008348 as an anti-fibrotic therapy

Mechanisms and models for industry engagement in collaborative research in commercial fisheries

Data and insights from fishers are essential sources of information to advance understanding of fishery and ecosystem dynamics. Incorporating fisher and industry knowledge holds prospects for improving marine science and fisheries management. We address cooperative research in the context of collaboration between fishers, scientists, industries, universities, and agencies to develop applied research to understand marine ecosystems, inform fishery management, enhance sustainability, govern resource use, and investigate social-economic dynamics. We leverage the insights of more than 100 research scientists, fisheries managers, industry representatives, and fishers to outline actionable recommendations for effective approaches and mechanisms to integrate industry data, perspectives, and insights in fisheries science. We also highlight opportunities and address challenges and limitations to such collaboration

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy