Anti-Red Fluorescent Protein monoclonal antibody, clone CW-G10

As an alternative to eye drops and ocular injections for gene therapy, the aim of this work was to design for the first time hydrogel contact lenses that can act as platforms for the controlled delivery of viral vectors (recombinant adeno-associated virus, rAAV) to the eye in an effective way with improved patient compliance. Hydrogels of hydroxyethyl methacrylate (HEMA) with aminopropyl methacrylamide (APMA) (H-1: 40, and H-2: 80 mM) or without (H-c: 0 mM) were synthesized, sterilized by steam heat (121 degrees C, 20 min), and then tested for gene therapy using rAAV vectors to deliver the genes to the cornea. The hydrogels showed adequate light transparency, oxygen permeability, and swelling for use as contact lenses. Loading of viral vectors (rAAV-lacZ, rAAV-RFP, or rAAV-hIGF-I) was carried out at 4 degrees C to maintain viral vector titer. Release in culture medium was monitored by fluorescence with Cy3-rAAV-lacZ and AAV Titration ELISA. Transduction efficacy was tested through reporter genes lacZ and RFP in human bone marrow derived mesenchymal stem cells (hMSCs). lacZ was detected with X-Gal staining and quantified with Beta-Glo((R)), and RFP was monitored by fluorescence. The ability of rAAV-hIGF-I-loaded hydrogels to trigger cell proliferation in hMSCs was evaluated by immunohistochemistry. Finally, the ability of rAAV-lacZ-loaded hydrogels to transduce bovine cornea was confirmed through detection with X-Gal staining of beta-galactosidase expressed within the tissue.

Objective The objective of this study was to analyze how marginalized men's coparenting perspectives and experiences shaped their engagement with a responsible fatherhood program (RFP). Background Previous studies of fathers enrolled in RFPs found that their coparenting relationships have both positive and negative components and that program participation is associated with better coparenting communication. Yet men also report strained coparenting relationships as a primary barrier to paternal involvement. More exploratory research is needed to understand how fathers use RFPs to navigate coparenting challenges. Method In-depth interviews and focus groups were conducted with a nonrandom sample of 64 low-income fathers of color who participated in a federally funded RFP. Inductive coding techniques revealed how men used the program to manage complex and conflicted coparenting arrangements and to gain greater access to their children. Results The program helped fathers manage common coparenting challenges, including interpersonal conflict, mothers' perceived gatekeeping, and financial obstacles. They found support for forming closer relationships with children outside "package deal" understandings of fathering and for proving their paternal commitments amid severe economic constraints. Conclusion Fathers enrolled in an RFP primarily due to conflicted coparenting relationships exacerbated by poverty, unemployment, and homelessness. Program services helped them navigate the challenges of raising children with partners, exes, or coparents with whom they never had a romantic connection. Implications These findings reinforce the importance of addressing coparenting in RFPs. Messages and services should align with how fathers prioritize bonds with children and seek to prove their paternal commitments to coparents.