Gene Transfer to Chicks Using Lentiviral Vectors Administered via the Embryonic Chorioallantoic Membrane

The lack of affordable techniques for gene transfer in birds has inhibited the advancement of molecular studies in avian species. Here we demonstrate a new approach for introducing genes into chicken somatic tissues by administration of a lentiviral vector, derived from the feline immunodeficiency virus (FIV), into the chorioallantoic membrane (CAM) of chick embryos on embryonic day 11. The FIV-derived vectors carried yellow fluorescent protein (YFP) or recombinant alpha-melanocyte-stimulating hormone (α-MSH) genes, driven by the cytomegalovirus (CMV) promoter. Transgene expression, detected in chicks 2 days after hatch by quantitative real-time PCR, was mostly observed in the liver and spleen. Lower expression levels were also detected in the brain, kidney, heart and breast muscle. Immunofluorescence and flow cytometry analyses confirmed transgene expression in chick tissues at the protein level, demonstrating a transduction efficiency of ∼0.46% of liver cells. Integration of the viral vector into the chicken genome was demonstrated using genomic repetitive (CR1)-PCR amplification. Viability and stability of the transduced cells was confirmed using terminal deoxynucleotidyl transferase (dUTP) nick end labeling (TUNEL) assay, immunostaining with anti-proliferating cell nuclear antigen (anti-PCNA), and detection of transgene expression 51 days post transduction. Our approach led to only 9% drop in hatching efficiency compared to non-injected embryos, and all of the hatched chicks expressed the transgenes. We suggest that the transduction efficiency of FIV vectors combined with the accessibility of the CAM vasculature as a delivery route comprise a new powerful and practical approach for gene delivery into somatic tissues of chickens. Most relevant is the efficient transduction of the liver, which specializes in the production and secretion of proteins, thereby providing an optimal target for prolonged study of secreted hormones and peptides

The James Webb Space Telescope Mission

Twenty-six years ago a small committee report, building on earlier studies, expounded a compelling and poetic vision for the future of astronomy, calling for an infrared-optimized space telescope with an aperture of at least $4m$. With the support of their governments in the US, Europe, and Canada, 20,000 people realized that vision as the $6.5m$ James Webb Space Telescope. A generation of astronomers will celebrate their accomplishments for the life of the mission, potentially as long as 20 years, and beyond. This report and the scientific discoveries that follow are extended thank-you notes to the 20,000 team members. The telescope is working perfectly, with much better image quality than expected. In this and accompanying papers, we give a brief history, describe the observatory, outline its objectives and current observing program, and discuss the inventions and people who made it possible. We cite detailed reports on the design and the measured performance on orbit.Comment: Accepted by PASP for the special issue on The James Webb Space Telescope Overview, 29 pages, 4 figure

Understanding the Working Alliance Between Persons with Substance Abuse Problems and Strengths-Based Case Managers

A substantial body of literature has examined the importance of the working alliance in psychotherapy; few works have examined it in the context of case management. Qualitative methods were used to examine how 26 persons with substance abuse problems perceived the working alliance with case managers who practice from the strengths perspective. Clients\u27 narratives emphasized the personal qualities of the case manager and the nature of the client-case manager relationship. Their narratives also reflected two guiding principles of strengths-based case management: personal control over goal-setting, and an emphasis on strengths. Most clients concluded that a positive working alliance helped them to build trust, self-worth, and self-esteem

Understanding the Working Alliance Between Persons with Substance Abuse Problems and Strengths-Based Case Managers

A substantial body of literature has examined the importance of the working alliance in psychotherapy; few works have examined it in the context of case management. Qualitative methods were used to examine how 26 persons with substance abuse problems perceived the working alliance with case managers who practice from the strengths perspective. Clients\u27 narratives emphasized the personal qualities of the case manager and the nature of the client-case manager relationship. Their narratives also reflected two guiding principles of strengths-based case management: personal control over goal-setting, and an emphasis on strengths. Most clients concluded that a positive working alliance helped them to build trust, self-worth, and self-esteem