Targeted gene delivery to the enteric nervous system using AAV: a comparison across serotypes and capsid mutants

Recombinant adeno-associated virus (AAV) vectors are one of the most widely used gene transfer systems in research and clinical trials. AAV can transduce a wide range of biological tissues, however to date, there has been no investigation on targeted AAV transduction of the enteric nervous system (ENS). Here, we examined the efficiency, tropism, spread, and immunogenicity of AAV transduction in the ENS. Rats received direct injections of various AAV serotypes expressing green fluorescent protein (GFP) into the descending colon. AAV serotypes tested included; AAV 1, 2, 5, 6, 8, or 9 and the AAV2 and AAV8 capsid mutants, AAV2-Y444F, AAV2-tripleY-F, AAV2-tripleY-F+T-V, AAV8-Y733F, and AAV8-doubeY-F+T-V. Transduction, as determined by GFP-positive cells, occurred in neurons and enteric glia within the myenteric and submucosal plexuses of the ENS. AAV6 and AAV9 showed the highest levels of transduction within the ENS. Transduction efficiency scaled with titer and time, was translated to the murine ENS, and produced no vector-related immune response. A single injection of AAV into the colon covered an area of ~47 mm(2). AAV9 primarily transduced neurons, while AAV6 transduced enteric glia and neurons. This is the first report on targeted AAV transduction of neurons and glia in the ENS

Chandra spectroscopy of the hot star beta Crucis and the discovery of a pre-main-sequence companion

In order to test the O star wind-shock scenario for X-ray production in less luminous stars with weaker winds, we made a pointed 74 ks observation of the nearby early B giant, beta Cru (B0.5 III), with the Chandra HETGS. We find that the X-ray spectrum is quite soft, with a dominant thermal component near 3 million K, and that the emission lines are resolved but quite narrow, with half-widths of 150 km/s. The forbidden-to-intercombination line ratios of Ne IX and Mg XI indicate that the hot plasma is distributed in the wind, rather than confined near the photosphere. It is difficult to understand the X-ray data in the context of the standard wind-shock paradigm for OB stars, primarily because of the narrow lines, but also because of the high X-ray production efficiency. A scenario in which the bulk of the outer wind is shock heated is broadly consistent with the data, but not very well motivated theoretically. It is possible that magnetic channeling could explain the X-ray properties, although no field has been detected on beta Cru. We detected periodic variability in the hard (hnu > 1 keV) X-rays, modulated on the known optical period of 4.58 hours, which is the period of the primary beta Cep pulsation mode for this star. We also have detected, for the first time, an apparent companion to beta Cru at a projected separation of 4 arcsec. This companion was likely never seen in optical images because of the presumed very high contrast between it and beta Cru in the optical. However, the brightness contrast in the X-ray is only 3:1, which is consistent with the companion being an X-ray active low-mass pre-main-sequence star. The companion's X-ray spectrum is relatively hard and variable, as would be expected from a post T Tauri star.Comment: Accepted for publication in MNRAS; 19 pages, 15 figures, some in color; version with higher-resolution figures available at http://astro.swarthmore.edu/~cohen/papers/bcru_mnras2008.pd

Trends in Compulsory Licensing of Pharmaceuticals Since the Doha Declaration: A Database Analysis

Reed Beall and Randall Kuhn describe their findings from an analysis of use of compulsory licenses for pharmaceutical products by World Trade Organization members since 1995

Training Models in Counseling Psychology: Scientist-Practitioner Versus Practitioner-Scholar

Considerable discussion has occurred through the years regarding models of training. With the recent accreditation of counseling psychology programs espousing the practitioner-scholar model, the importance of reexamining the merits of this as well as the traditional scientist-practitioner is now very important for the future of the field. This article consists of two positions: One pro practitioner-scholar and the other pro scientist-practitioner and con practitioner-scholar. The first position (first part of the article) by Biever, Patterson, and Welch argues for inclusion of the practitioner-scholar model as an alternative for training in counseling psychology. The second position (in the second part of the article) by Stoltenberg, Pace, and Kashubeck reviews concerns with two competing models. These authors conclude that the scientist-practitioner model is a better fit for training in counseling psychology. Recommendations for training within models are presented.Yeshttps://us.sagepub.com/en-us/nam/manuscript-submission-guideline

A Practical Platform for Blood Biomarker Study by Using Global Gene Expression Profiling of Peripheral Whole Blood

Background: Although microarray technology has become the most common method for studying global gene expression, a plethora of technical factors across the experiment contribute to the variable of genome gene expression profiling using peripheral whole blood. A practical platform needs to be established in order to obtain reliable and reproducible data to meet clinical requirements for biomarker study. Methods and Findings: We applied peripheral whole blood samples with globin reduction and performed genome-wide transcriptome analysis using Illumina BeadChips. Real-time PCR was subsequently used to evaluate the quality of array data and elucidate the mode in which hemoglobin interferes in gene expression profiling. We demonstrated that, when applied in the context of standard microarray processing procedures, globin reduction results in a consistent and significant increase in the quality of beadarray data. When compared to their pre-globin reduction counterparts, post-globin reduction samples show improved detection statistics, lowered variance and increased sensitivity. More importantly, gender gene separation is remarkably clearer in post-globin reduction samples than in pre-globin reduction samples. Our study suggests that the poor data obtained from pre-globin reduction samples is the result of the high concentration of hemoglobin derived from red blood cells either interfering with target mRNA binding or giving the pseudo binding background signal. Conclusion: We therefore recommend the combination of performing globin mRNA reduction in peripheral whole blood samples and hybridizing on Illumina BeadChips as the practical approach for biomarker study

Postnatal PPARδ Activation and Myostatin Inhibition Exert Distinct yet Complimentary Effects on the Metabolic Profile of Obese Insulin-Resistant Mice

BACKGROUND: Interventions for T2DM have in part aimed to mimic exercise. Here, we have compared the independent and combined effects of a PPARdelta agonist and endurance training mimetic (GW501516) and a myostatin antibody and resistance training mimetic (PF-879) on metabolic and performance outcomes in obese insulin resistant mice. METHODOLOGY/PRINCIPAL FINDINGS: Male ob/ob mice were treated for 6 weeks with vehicle, GW501516, PF-879, or GW501516 in combination with PF-879. The effects of the interventions on body composition, glucose homeostasis, glucose tolerance, energy expenditure, exercise capacity and metabolic gene expression were compared at the end of study. GW501516 attenuated body weight and fat mass accumulation and increased the expression of genes of oxidative metabolism. In contrast, PF-879 increased body weight by driving muscle growth and altered the expression of genes involved in insulin signaling and glucose metabolism. Despite their differences, both interventions alone improved glucose homeostasis. Moreover, GW501516 more effectively improved serum lipids, and PF-879 uniquely increased energy expenditure, exercise capacity and adiponectin levels. When combined the robust effects of GW501516 and/or PF-879 on body weight, adiposity, muscle mass, glycemia, serum lipids, energy expenditure and exercise capacity were highly conserved. CONCLUSIONS/SIGNIFICANCE: The data, for the first time, demonstrate postnatal inhibition of myostatin not only promotes gains in muscle mass similar to resistance training,but improves metabolic homeostasis. In several instances, these effects were either distinct from or complimentary to those of GW501516. The data further suggest that strategies to increase muscle mass, and not necessarily oxidative capacity, may effectively counter insulin resistance and T2DM

Nomenclature- and Database-Compatible Names for the Two Ebola Virus Variants that Emerged in Guinea and the Democratic Republic of the Congo in 2014

In 2014, Ebola virus (EBOV) was identified as the etiological agent of a large and still expanding outbreak of Ebola virus disease (EVD) in West Africa and a much more confined EVD outbreak in Middle Africa. Epidemiological and evolutionary analyses confirmed that all cases of both outbreaks are connected to a single introduction each of EBOV into human populations and that both outbreaks are not directly connected. Coding-complete genomic sequence analyses of isolates revealed that the two outbreaks were caused by two novel EBOV variants, and initial clinical observations suggest that neither of them should be considered strains. Here we present consensus decisions on naming for both variants (West Africa: “Makona”, Middle Africa: “Lomela”) and provide database-compatible full, shortened, and abbreviated names that are in line with recently established filovirus sub-species nomenclatures

Effectiveness of Non-nucleoside Reverse-Transcriptase Inhibitor-Based Antiretroviral Therapy in Women Previously Exposed to a Single Intrapartum Dose of Nevirapine: A Multi-country, Prospective Cohort Study

In a comparative cohort study, Jeffrey Stringer and colleagues investigate the risk of ART failure in women who received single-dose nevirapine for PMTCT, and assess the duration of increased risk