The in vivo and in vitro formation of sticky DNA and the GAA.TTC trinucleotide repeat associated with Friedreich's ataxia

Friedreich's ataxia is caused by the expansion of the GAA•TTC trinucleotide repeat sequence located in intron 1 of the frataxin gene. The long GAA•TTC repeats are known to form several non-B DNA structures including hairpins, triplexes, parallel DNA and sticky DNA. Therefore it is believed that alternative DNA structures play a role in the loss of mRNA transcript and functional frataxin protein in FRDA patients. We wanted to further elucidate the characteristics for formation and stability of sticky DNA by evaluating the structure in a plasmid based system in vitro and in vivo in Escherichia coli. The negative supercoil density of plasmids harboring different lengths of GAA•TTC repeats, as well as either one or two repeat tracts were studied in E. coli to determine if plasmids containing two long tracts (≥60 repeats) in a direct repeat orientation would have a different topological effect in vivo compared to plasmids that harbored only one GAA•TTC tract or two tracts of < 60 repeats. The experiments revealed that, in fact, sticky DNA forming plasmids had a lower average negative supercoil density (-σ) compared to all other control plasmids used that had the potential to form other non-B DNA structures such as triplexes or Z-DNA. Also, the requirements for in vitro dissociation and reconstitution of the DNA•DNA associated region of sticky DNA were evaluated. Results conclude that the two repeat tracts associate in the presence of negative supercoiling and MgCl 2 or MnCl2 in a time and concentration-dependent manner. Interaction of the repeat sequences was not observed in the absence of negative supercoiling and/or MgCl2 or in the presence of other monovalent or divalent cations, indicating that supercoiling and quite specific cations are needed for the association of sticky DNA. These are the first experiments studying a more specific role of supercoiling and cation influence on this DNA conformation. To support our model of the topological effects of sticky DNA in plasmids, changes in sticky DNA band migration was measured with reference to the linear DNA after treatment with increasing concentrations of ethidium bromide (EtBr). The presence of independent negative supercoil domains was confirmed by this method and found to be segregated by the DNA-DNA associated region. Sequence-specific polyamide molecules were used to test the effect of binding of the ligands to the GAA•TTC repeats on the inhibition of sticky DNA. The destabilization of the sticky DNA conformation in vitro through this binding of the polyamides demonstrated the first conceptual therapeutic approach for the treatment of FRDA at the DNA molecular level. ^ Thus, examining the properties of sticky DNA formed by these long repeat tracts is important in the elucidation of the possible role of sticky DNA in Friedreich's ataxia. ^

Analysis of community health worker utilization in El Paso County, Texas

Community health workers (CHWs) are an accepted, viable component of health systems worldwide. Most often they are utilized in developing regions where health care access is limited and/or health care practitioners are scarce. In this way community health workers have been used to extend the reach of primary care delivery to whole nations, and can be used to reduce health disparities in disadvantaged populations or minority groups as well. In the United States, utilization of CHWs is fragmented, and an amalgam of programs exist which are usually only community-specific. These programs are often burdened by financial un-sustainability. The Community Health Worker National Workforce Study (2007) was markedly the first effort to compile a profile of the CHW workforce in all 50 states. El Paso County, as a uniquely bi-national setting, provided a prime locale to assess CHW utilization on a localized scale, and in a distinctively Latino population and medically underserved area. ^ Results gleaned from this study of 45 CHWs and 5 administrators demonstrate commonalities between El Paso County CHWs and the national CHW workforce; differences were found in average education level, wages for experienced CHWs, as well as primary target populace and target health issues. Future research should focus on cost-effectiveness of CHW utilization.^

Minimum income for healthy living

Objectives: We sought to estimate the costs of implementing the current recommendations for healthy choices for a mother with two young children in Atlanta, Georgia. ^ Methods: Current recommendations for healthy choices promoted by the federal government or other credible source were compiled and operationalized into specific conditions or behaviors. The costs of implementing these choices in Atlanta were estimated by using internet searches of retailers/suppliers, phone interviews, and direct observation. The least expensive option was chosen when options were available. ^ Results: Recommendations for choosing a healthy neighborhood, home, school, child care, food, physical activity, and maintaining healthy relationships as well as access to health care were considered. Total costs for this family of three totaled $38,181. Housing, child care, and health insurance contributed to 78% of the total costs. ^ Conclusions: The minimum income needed to choose healthy choices falls short of current wages, and eligibility levels and benefits for income support. ^