Women Leading to Make a Difference: An Inside Look at a Strength-based Home Visiting Program in Rural Appalachia

The Maternal Infant Outreach Worker Program (MIHOW) is a strength-based home visitation program that uses trained lay women indigenous to the community to mentor and teach parents who are economically disadvantaged or live in geographically isolated areas about healthy and positive pregnancy and parenting up until the child turns age three. This qualitative case study conducted in rural Appalachia at two program sites examined how women involved in the West Virginia MIHOW program – program leaders, home visitors, and mothers – came to recognize their strengths and use them to achieve life aspirations. In addition, this study explored how MIHOW program participants perceived themselves in various aspects of their lives and how the program contributed to positive social change for women, their families, and their communities. Findings were interpreted in relation to extant literature on strength-based approaches, home visitation, and women as leaders. Theme one pertains to the role of the importance of being explicit about strengths and making it pervasive throughout the entire program. Recognizing strengths and carrying out the strength-based approach was core for MIHOW program leaders and home visitors as they wholeheartedly practiced it in their work and their lives, whereas mothers’ recognition of their strengths was less clear. The second theme shows that MIHOW program staff and mothers achieved many of their life aspirations, as well as established new visions and overcame obstacles. The third theme shows that women participating in MIHOW were making a difference by simultaneously leading from in front (as role models) and from beside (as collaborative team members), which included the factors of authentically walking the walk of the strength-based approach, listening and observing with an open mind, collaborating with humility, and advocating for and with mothers. Findings were also interpreted through an examination of Robert K. Greenleaf’s servant leadership principles and the theoretical frame of social justice feminism. The combination of Robert K. Greenleaf’s (2002) servant leadership and social justice feminism was exemplified in MIHOW’s leadership from in front and from beside as it provided a respectful, supportive, encouraging, and egalitarian environment, which for many program staff and mothers increased their self-advocacy beliefs, fostered their leadership growth, empowered them to be the “leaders they wanted to be,” and transformed them into “movers and shakers” in their communities

Cosmic ray diffusive acceleration at shock waves with finite upstream and downstream escape boundaries

In the present paper we discuss the modifications introduced into the first-order Fermi shock acceleration process due to a finite extent of diffusive regions near the shock or due to boundary conditions leading to an increased particle escape upstream and/or downstream the shock. In the considered simple example of the planar shock wave we idealize the escape phenomenon by imposing a particle escape boundary at some distance from the shock. Presence of such a boundary (or boundaries) leads to coupled steepening of the accelerated particle spectrum and decreasing of the acceleration time scale. It allows for a semi-quantitative evaluation and, in some specific cases, also for modelling of the observed steep particle spectra as a result of the first-order Fermi shock acceleration. We also note that the particles close to the upper energy cut-off are younger than the estimate based on the respective acceleration time scale. In Appendix A we present a new time-dependent solution for infinite diffusive regions near the shock allowing for different constant diffusion coefficients upstream and downstream the shock.Comment: LaTeX, 14 pages, 4 postscript figures; Solar Physics (accepted

Proliferation of genetically modified human cells on electrospun nanofiber scaffolds.

Gene editing is a process by which single base mutations can be corrected, in the context of the chromosome, using single-stranded oligodeoxynucleotides (ssODNs). The survival and proliferation of the corrected cells bearing modified genes, however, are impeded by a phenomenon known as reduced proliferation phenotype (RPP); this is a barrier to practical implementation. To overcome the RPP problem, we utilized nanofiber scaffolds as templates on which modified cells were allowed to recover, grow, and expand after gene editing. Here, we present evidence that some HCT116-19, bearing an integrated, mutated enhanced green fluorescent protein (eGFP) gene and corrected by gene editing, proliferate on polylysine or fibronectin-coated polycaprolactone (PCL) nanofiber scaffolds. In contrast, no cells from the same reaction protocol plated on both regular dish surfaces and polylysine (or fibronectin)-coated dish surfaces proliferate. Therefore, growing genetically modified (edited) cells on electrospun nanofiber scaffolds promotes the reversal of the RPP and increases the potential of gene editing as an ex vivo gene therapy application.Molecular Therapy - Nucleic Acids (2012) 1, e59; doi:10.1038/mtna.2012.51; published online 4 December 2012

A stress-free strategy to correct point mutations in patient iPS cells

When studying patient specific induced pluripotent stem cells (iPS cells) as a disease model, the ideal control is an isogenic line that has corrected the point mutation, instead of iPS cells from siblings or other healthy subjects. However, repairing a point mutation in iPS cells even with the newly developed CRISPR-Cas9 technique remains difficult and time-consuming. Here we report a strategy that makes the Cas9 “knock-in” methodology both hassle-free and error-free. Instead of selecting a Cas9 recognition site close to the point mutation, we chose a site located in the nearest intron. We constructed a donor template with the fragment containing the corrected point mutation as one of the homologous recombination arms flanking a PGK-PuroR cassette. After selection with puromycin, positive clones were identified and further transfected with a CRE vector to remove the PGK-PuroR cassette. Using this methodology, we successfully repaired the point mutation G2019S of the LRRK2 gene in a Parkinson Disease (PD) patient iPS line and the point mutation R329H of the AARS1 gene in a Charcot-Marie-Tooth disease (CMT) patient iPS line. These isogenic iPS lines are ideal as a control in future studies

An efficient and scalable pipeline for epitope tagging in mammalian stem cells using Cas9 ribonucleoprotein

AbstractCRISPR/Cas9 can be used for precise genetic knock-in of epitope tags into endogenous genes, simplifying experimental analysis of protein function. However, Cas9-assisted epitope tagging in primary mammalian cell cultures is often inefficient and reliant on plasmid-based selection strategies. Here we demonstrate improved knock-in efficiencies of diverse tags (V5, 3XFLAG, Myc, HA) using co-delivery of Cas9 protein pre-complexed with two-part synthetic modified RNAs (annealed crRNA:tracrRNA) and single-stranded oligodeoxynucleotide (ssODN) repair templates. Knock-in efficiencies of ~5-30%, were achieved without selection in embryonic stem (ES) cells, neural stem (NS) cells, and brain tumour-derived stem cells. Biallelic-tagged clonal lines were readily derived and used to define Olig2 chromatin-bound interacting partners. Using our novel web-based design tool, we established a 96-well format pipeline that enabled V5-tagging of sixty different transcription factors. This efficient, selection-free and scalable epitope tagging pipeline enables systematic surveys of protein expression levels, subcellular localization, and interactors across diverse mammalian stem cells.</jats:p

Sorption and Photodegradation Processes Govern Distribution and Fate of Sulfamethazine in Freshwater−Sediment Microcosms

The antibiotic sulfamethazine can be transported from manured fields to surface water bodies. We investigated the degradation and fate of sulfamethazine in pond water using 14C-phenyl-sulfamethazine in small pond water microcosms containing intact sediment and pond water. We found a 2.7-day half-life in pond water and 4.2-day half-life when sulfamethazine was added to the water (5 mg L–1 initial concentration) with swine manure diluted to simulate runoff. Sulfamethazine dissipated exponentially from the water column, with the majority of loss occurring via movement into the sediment phase. Extractable sulfamethazine in sediment accounted for 1.9–6.1% of the applied antibiotic within 14 days and then declined thereafter. Sulfamethazine was transformed mainly into nonextractable sediment-bound residue (40–60% of applied radioactivity) and smaller amounts of photoproducts. Biodegradation, as indicated by metabolite formation and 14CO2 evolution, was less significant than photodegradation. Two photoproducts accounted for 15–30% of radioactivity in the water column at the end of the 63-day study; the photoproducts were the major degradates in the aqueous and sediment phases. Other unidentified metabolites individually accounted for \u3c7% of radioactivity in the water or sediment. Less than 3% of applied radioactivity was mineralized to 14CO2. Manure input significantly increased sorption and binding of sulfamethazine residues to the sediment. These results show concurrent processes of photodegradation and sorption to sediment control aqueous concentrations and establish that sediment is a sink for sulfamethazine and sulfamethazine-related residues. Accumulation of the photoproducts and sulfamethazine in sediment may have important implications for benthic organisms