Design and evaluation of the parent education component of a comprehensive violence prevention project in middle schools

Low parental monitoring is related to youth risk behaviors such as delinquency and aggression. The purpose of this dissertation was to describe the development and evaluation of a parent education intervention to increase parental monitoring in Hispanic parents of middle school children.^ The first study described the process of intervention mapping as used to develop Padres Trabajando por la Paz, a newsletter intervention for parents. Using theory, empirical literature, and information from the target population, performance objectives and determinants for monitoring were defined. Learning objectives were specified and a staged social-cognitive approach was used to develop methods and strategies delivered through newsletters.^ The second study examined the outcomes of a randomized trial of the newsletter intervention. Outcome measures consisted of a general measure of monitoring, parent and child reports of monitoring behaviors targeted by the intervention, and psychosocial determinants of monitoring (self-efficacy, norms, outcome expectancies, knowledge, and beliefs). Seventy-seven parents completed the randomized trial, half of which received four newsletters over an eight-week period. Results revealed a significant interaction effect for baseline and treatment for parent's reports of norms for monitoring (p =.009). Parents in the experimental condition who scored low at baseline reported increased norms for monitoring at follow-up. A significant interaction effect for child reports of parental monitoring behaviors (p =.04) reflected an small increase across baseline levels in the experimental condition and decreases for the control condition at higher baseline scores. Both groups of parents reported increased levels of monitoring at follow-up. No other outcome measures varied significantly by condition.^ The third study examined the relationship between the psychosocial determinants of parental monitoring and parental monitoring behaviors in the study population. Weak evidence for a relationship between outcome expectancies and parental monitoring behaviors suggests further research in the area utilizing stronger empirical models such as longitudinal design and structural equation modeling.^ The low-cost, minimal newsletter intervention showed promise for changing norms among Hispanic parents for parental monitoring. In light of the importance of parental monitoring as a protective factor for youth health risk behaviors, more research needs to be done to develop and evaluate interventions to increase parental monitoring. ^

NMR structural analysis of cardiac troponin C: Monitoring conformational changes induced by binding calcium, troponin I, and calcium -sensitizing drugs

Contraction of cardiac muscle is regulated through the Ca2+ dependent protein-protein interactions of the troponin complex (Tn). The critical role cardiac troponin C (cTnC) plays as the Ca2+ receptor in this complex makes it an attractive target for positive inotropic compounds. In this study, the ten Met methyl groups in cTnC, [98% 13C ϵ]-Met cTnC, are used as structural markers to monitor conformational changes in cTnC and identify sites of interaction between cTnC and cardiac troponin I (cTnI) responsible for the Ca2+ dependent interactions. In addition the structural consequences that a number of Ca2+-sensitizing compounds have on free cTnC and the cTnC·cTnI complex were characterized. Using heteronuclear NMR experiments and monitoring chemical shift changes in the ten Met methyl 1H-13C correlations in 3Ca2+ cTnC when bound to cTnI revealed an anti-parallel arrangement for the two proteins such that the N-domain of cTnI interacts with the C-domain of cTnC. The large chemical shifts in Mets-81, -120, and -157 identified points of contact between the proteins that include the C-domain hydrophobic surface in cTnC and the A, B, and D helical interface located in the regulatory N-domain of cTnC. TnI association [cTnI(33–80), cTnI(86–211), or cTnI(33–211)] was found also to dramatically reduce flexibility in the D/E central linker of cTnC as monitored by line broadening in the Met 1H- 13C correlations of cTnC induced by a nitroxide spin label, MTSSL, covalently attached to cTnC at Cys 84. TnI association resulted in an extended cTnC that is unlike the compact structure observed for free cTnC. The Met 1H-13C correlations also allowed the binding characteristics of bepridil, TFP, levosimendan, and EMD 57033 to the apo, 2Ca2+, and Ca2+ saturated forms of cTnC to be determined. In addition, the location of drug binding on the 3Ca2+cTnC·cTnI complex was identified for bepridil and TFP. Use of a novel spin-labeled phenothiazine, and detection of isotope filtered NOEs, allowed identification of drug binding sites in the shallow hydrophobic cup in the C-terminal domain, and on two hydrophobic surfaces on N-regulatory domain in free 3Ca2+ cTnC. In contrast, only one N-domain drug binding site exists in 3Ca2+ cTnC·cTnI complex. The methyl groups of Met 45, 60 and 80, which are grouped in a hydrophobic patch near site II in cTnC, showed the greatest change upon titration with bepridil or TFP, suggesting that this is a critical site of drug binding in both free cTnC and when associated with cTnI. The strongest NOEs were seen for Met-60 and -80, which are located on helices C and D, respectively, of Ca2+ binding site II. These results support the conclusion that the small hydrophobic patch which includes Met-45, -60, and -80 constitutes a drug binding site, and that binding drugs to this site will lead to an increase in Ca2+ binding affinity of site II while preserving maximal cTnC activity. Thus, the subregion in cTnC makes a likely target against which to design new and selective Ca2+-sensitizing compounds. ^