Genetic Correction of Dystrophin Deficiency and Skeletal Muscle Remodeling in Adult MDX Mouse via Transplantation of Retroviral Producer Cells

Duchenne muscular dystrophy (DMD) is an X-linked, lethal disease caused by mutations of the dystrophin gene. No effective therapy is available, but dystrophin gene transfer to skeletal muscle has been proposed as a treatment for DMD. We have developed a strategy for efficient in vivo gene transfer of dystrophin cDNA into regenerating skeletal muscle. Retroviral producer cells, which release a vector carrying the therapeutically active dystrophin minigene, were mitotically inactivated and transplanted in adult nude/ mdx mice. Transplantation of 3 3 10 6 producer cells in a single site of the tibialis anterior muscle resulted in the transduction of between 5.5 and 18% total muscle fibers. The same procedure proved also feasible in immunocompetent mdx mice under short-term pharmacological immunosuppression. Minidystrophin expression was stable for up to 6 mo and led to a -sarcoglycan reexpression. Muscle stem cells could be transduced in vivo using this procedure. Transduced dystrophic skeletal muscle showed evidence of active remodeling reminiscent of the genetic normalization process which takes place in female DMD carriers. Overall, these results demonstrate that retroviral-mediated dystrophin gene transfer via transplantation of producer cells is a valid approach towards the long-term goal of gene therapy of DMD. ( J. Clin. Invest. 1997. 100:620–628 .

LAYING THE FOUNDATION FOR SELF-MANAGEMENT SUPPORT IN A RECOVERY FRAMEWORK

For more information contact: Susan Strong, [email protected]; St Joseph's Healthcare Hamilton, West 5th Campus, Schizophrenia & Community Integration Service, 100 West 5th St., Hamilton, ON L8N3K7Introduction Despite international pressure to implement self-management support for individuals living with chronic illnesses, little direction is available for integration into specialized mental health services. The premise of the dissertation is that self-management support can be provided and be beneficial within a recovery framework for individuals living with serious mental illnesses. Methods A large regional service was studied as an exemplar of specialized service delivery. Using a van Manen phenomenological study through an occupational therapist lens, the meaning of clients’ experiences with self-management learning events in a diverse sample was examined. Conditions shaping clinicians’ experiences and actions enabling self-management across eight varied settings were identified in a Yin case study with embedded units. Provider triads (occupational therapist-nurse-social worker) from each setting, allowed an examination of patterns by individual, discipline and practice environment. With an integrated knowledge translation approach, both studies informed the strategic creation of an innovation and implementation plan for organizational change. Results Findings created a rich picture of clients’ lifeworlds learning self-management and conditions and mechanisms influencing clinician self-management support practices. Client self-management needs were not routinely addressed by services. Clients experienced eight tasks shaped by contextual structures represented in a model of the work of learning self-management. Self-management was enmeshed in recovery and a personal resource for self-determination and living well. Another model illustrated the complex dynamic relationships underpinning clinicians’ intentions and actions, and key features of clinicians enabling client self-management. A structured approach to delivering self-management support and a learning and embedding initiative were generated. Conclusions The dominance of practice environments offers an opportunity for organizational change. Direction is given to customize self-management support in a conventional mental health service. The conceptualization of self-management support within a recovery framework may benefit all client groups with long-standing conditions.Doctor of Philosophy (PhD

CH 3 GHz Observations of the Galactic Center

A 3 $\times$ 3 map of the Galactic Center was made at 9\arcmin resolution and 10\arcmin spacing in the CH $^2\Pi_{1/2}$, J=1/2, F=1-1 transition at 3335 MHz. The CH emission shows a velocity extent that is nearly that of the CO(1-0) line, but the CH line profiles differ markedly from the CO. The 3335 MHz CH transition primarily traces low-density molecular gas and our observations indicate that the mass of this component within $\sim$ 30 pc of the Galactic Center is $\sim$ 9 $\times$ 10$^6$ M$_\odot$. The CO-H$_2$ conversion factor obtained for the low-density gas in the mapped region is greater than that thought to apply to the dense molecular gas at the Galactic Center. In addition to tracing the low-density molecular gas at the Galactic Center, the CH spectra show evidence of emission from molecular clouds along the line of sight both in the foreground and background. The scale height of these clouds ranges from 27 - 109 pc, consistent with previous work based on observations of molecular clouds in the inner Galaxy.Comment: 29 pages, 12 figure

Translational profiling of hypocretin neurons identifies candidate molecules for sleep regulation

Hypocretin (orexin; Hcrt)-containing neurons of the hypothalamus are essential for the normal regulation of sleep and wake behaviors and have been implicated in feeding, anxiety, depression, and reward. The absence of these neurons causes narcolepsy in humans and model organisms. However, little is known about the molecular phenotype of these cells; previous attempts at comprehensive profiling had only limited sensitivity or were inaccurate. We generated a Hcrt translating ribosome affinity purification (bacTRAP) line for comprehensive translational profiling of all ribosome-bound transcripts in these neurons in vivo. From this profile, we identified >6000 transcripts detectably expressed above background and 188 transcripts that are highly enriched in these neurons, including all known markers of the cells. Blinded analysis of in situ hybridization databases suggests that ∼60% of these are expressed in a Hcrt marker-like pattern. Fifteen of these were confirmed with double labeling and microscopy, including the transcription factor Lhx9. Ablation of this gene results in a >30% loss specifically of Hcrt neurons, without a general disruption of hypothalamic development. Polysomnography and activity monitoring revealed a profound hypersomnolence in these mice. These data provide an in-depth and accurate profile of Hcrt neuron gene expression and suggest that Lhx9 may be important for specification or survival of a subset of these cells

Identification and characterization of glycoproteins on the spore surface of Clostridium difficile

In this study, we identify a major spore surface protein, BclA, and provide evidence that this protein is glycosylated. Following extraction of the spore surface, solubilized proteins were separated by one-dimensional PAGE and stained with glycostain to reveal a reactive high-molecular-mass region of approximately 600 kDa. Tandem mass spectrometry analysis of in-gel digests showed this band to contain peptides corresponding to a putative exosporangial glycoprotein (BclA3) and identified a number of glycopeptides modified with multiple N-acetyl hexosamine moieties and, in some cases, capped with novel glycans. In addition, we demonstrate that the glycosyltransferase gene sgtA (gene CD3350 in strain 630 and CDR3194 in strain R20291), which is located immediately upstream of the bclA3 homolog, is involved in the glycosylation of the spore surface, and is cotranscribed with bclA3. The presence of anti-β-O-GlcNAc-reactive material was demonstrated on the surface of spores by immunofluorescence and in surface extracts by Western blotting, although each strain produced a distinct pattern of reactivity. Reactivity of the spore surface with the anti-β-O-GlcNAc antibody was abolished in the 630 and R20291 glycosyltransferase mutant strains, while complementation with a wild-type copy of the gene restored the β-O-GlcNAc reactivity. Phenotypic testing of R20291 glycosyltransferase mutant spores revealed no significant change in sensitivity to ethanol or lysozyme. However, a change in the resistance to heat of R20291 glycosyltransferase mutant spores compared to R20291 spores was observed, as was the ability to adhere to and be internalized by macrophages

Examining the Interface Between Substance Misuse and Intimate Partner Violence

There is considerable theoretical and empirical support for a link between substance misuse and perpetration and victimization of intimate partner violence. This review briefly summarizes this literature and highlights current research that addresses the interface between treatment for substance abuse and intimate partner violence. Suggestions for future research and clinical implications are provided

In Support of a Patient-Driven Initiative and Petition to Lower the High Price of Cancer Drugs

Comment in Lowering the High Cost of Cancer Drugs--III. [Mayo Clin Proc. 2016] Lowering the High Cost of Cancer Drugs--I. [Mayo Clin Proc. 2016] Lowering the High Cost of Cancer Drugs--IV. [Mayo Clin Proc. 2016] In Reply--Lowering the High Cost of Cancer Drugs. [Mayo Clin Proc. 2016] US oncologists call for government regulation to curb drug price rises. [BMJ. 2015

Waterbird monitoring and habitat association modeling to inform tidal marsh restoration in an urbanized estuary

The San Francisco Bay (SFB), like many other urbanized estuaries, is a critical wintering and stop-over area for migratory waterbirds. More than a million wintering waterbirds annually rely on a mosaic of natural and managed habitats in SFB, including former salt ponds. The South Bay Salt Pond Restoration Project plans restore 50 to 90% of a 6,100 hectare former salt production pond complex to tidal marsh, while maintaining the rest as foraging and roosting areas for migratory birds. Since 2002, we have evaluated migratory waterbird use of pre- and post-restoration salt ponds in south SFB. Our approach has been to use monthly surveys, applied studies and modeling to evaluate avian response to this changing habitat. Trend analyses indicate that overall waterbird numbers have nearly doubled across the complex from 2003-2014. Dabbling ducks increased significantly during initial phases of the project, while diving ducks, small and medium shorebird abundances declined initially, but rebounded in recent years. We modeled relationships between abundances of multiple species and habitat characteristics at both landscape and micro-scales. Modeling results suggest that the importance of different habitat characteristics varies among waterbird guilds. For example, foraging diving duck abundances were higher in deep, un-breached ponds, located close to the edge of SFB, while foraging small shorebirds predominated in shallow, breached ponds containing islands. Our research highlights important habitat characteristics both within and across wetland areas, and provides a unique opportunity to guide future management and restoration decisions for multiple species across large landscapes in SFB and other urbanized estuaries

Diet, Weight Management, Physical activity and Ovarian & Breast Cancer Risk in Women With

INTRODUCTION: Women with pathogenic germline gene variants in METHODS: We searched Medline, EMBASE, CENTRAL, PubMed, and clinicaltrials.gov up to October 3, 2019. We identified 2775 records and included 21. RESULTS: There is limited evidence related to these factors and ovarian cancer risk. For breast cancer risk, evidence suggests higher diet quality, adulthood weight-loss of ≥10 pounds, and activity during adolescence and young-adulthood may be linked with decreased risk. Higher meat intake and higher daily energy intake may be linked with increased risk. CONCLUSIONS: There is not enough evidence to suggest tailored recommendations for dietary habits or weight management among women wit