Publications

Regulation of GH gene expression by GRF involves cAMP as a second messenger. We have demonstrated that a 500-basepair fragment of the human GH (hGH) gene 5' flanking region can confer cAMP inducibility upon the chloramphenicol acetyltransferase transcription unit in transient transfections of rat pituitary tumor cells treated with forskolin, an activator of adenyl cyclase. The same hGH construct is not induced by forskolin in nonpituitary-derived cells. Experiments with hGH deletion constructs reveal that binding sites for transcription factor AP-2 and the pituitary-specific factor GHF-1 are not required for forskolin stimulation, but that GHF-1 may potentiate the effect. RNA analyses reveal that forskolin also stimulates accumulation of transcripts initiated at the hGH promoter. Other agents that elevate cAMP levels also stimulate hGH expression. Since the hGH 5' flanking region contains no sequences homologous to the cAMP-responsive element of the somatostatin gene, and the AP-2 sites do not appear to be required for the forskolin response, these results suggest that a novel cAMP-responsive element exists within 82 basepairs upstream from the transcriptional start of the hGH gene and that hGH regulation by GRF may involve interaction between a tissue-specific element and a cAMP-inducible element.

Activation of either muscarinic cholinergic or thrombin receptors increases phosphoinositide turnover, Ca2+ mobilization, and redistribution of protein kinase C and induces rapid transient increases in c-fos mRNA and c-jun mRNA in 1321N1 cells. To determine whether the increases in c-fos and c-jun mRNA induced by carbachol and thrombin are sufficient to stimulate AP-1-mediated transactivation, 1321N1 cells were transfected with a reporter carrying two copies of the tetradecanoyl phorbol acetate response element and the firefly luciferase gene. Thrombin was significantly more effective than carbachol at stimulating AP-1-mediated transactivation. To identify the factors underlying the difference in AP-1 activity induced by carbachol and thrombin, members of the fos and jun families which encode components of AP-1 were examined. Carbachol and thrombin have similar effects on expression of c-fos, fosB, fra-2, junB, and junD, both acutely and over a 24-h time course. However, whereas carbachol leads only to transient induction of c-jun (maximal at 0.5 h), thrombin induces a biphasic increase in c-jun mRNA--an initial peak at 0.5 h and a second, more-prolonged increase at 12 h. Thrombin but not carbachol also induces a late increase in fra-1 mRNA, which peaks at 12 h. The secondary increase in c-jun mRNA is associated with marked increases in c-Jun protein levels and AP-1 DNA-binding activity. The late induction of c-jun and fra-1 mRNA can be prevented by adding the antagonist hirudin 30 min after thrombin, which results in loss of thrombin-stimulated increases in c-Jun protein, AP-1 DNA-binding activity, and AP-1-mediated transactivation. These findings suggest that rapid and transient conduction of c-fos and c-jun mRNA is insufficient to induce prominent changes in gene transcription, while the sustained increase in c-jun mRNA and perhaps the late induction of fra-1 mRNA are required for generation of AP-1 DNA-binding activity and transactivation through AP-1.

This study assessed the comparability and reliability of paper-based and computer-based administration of psychosocial construct measures related to adolescents engaging in physical activity and sedentary behaviors. Adolescents (n = 76; 55% girls, 49% Hispanic, 18% non-Hispanic-white; M age = 13 years) completed measures of behavior change strategies, selfefficacy, decisional balance, family and peer influences, enjoyment, activity choices, and environmental influences. Overall, the adolescents provided equivalent responses on paper and computer formats. Reliability estimates were generally good for the multiple-item constructs, but single-item measures tended to demonstrate low reliability. Average scale internal consistency was significantly higher for measures administered with the computer format compared to the paper format, but test-rest reliability estimates were not significantly different between formats.

It is hypothesized that high levels of hostility stimulate blood pressure hyperresponsivity to challenge. Seventy-six normotensive adults completed the MMPI hostility subscale, and blood pressure reactivity to, and recovery from, cold pressor and mental arithmetic stressors were measured on two occasions. Only one of twenty Pearson correlations between Cook-Medley hostility and blood pressure reactivity was significant. Analyses of variance revealed that blood pressure reactivity was influenced by sex and family history of cardiovascular disease, but neither factor interacted with hostility. There was no evidence from the present study that cynical hostility as measured by the Cook-Medley was associated with blood pressure reactivity.

Classroom teachers’ responses to a 2-year professional development program are presented. Sixteen 4th- and 5th-grade teachers involved in Project SPARK completed structured interviews, questionnaires, and written evaluations of program sessions. Although in Year 1 more than half of the teachers expressed concerns about schedules and equipment management, results indicated that the program helped increase their self-confidence when teaching physical education. Participants believed that students benefitted from their enhanced knowledge and instructional behaviors. Program components most appreciated included: the input received and responsiveness of the design team; opportunities to collaborate, discuss concerns, and problem-solve with each other and the facilitators; and having on-site and large-group-session modeling. Results indicated that the teachers were less enthusiastic about a self-management curriculum due to its behavioral emphasis, yet supported the assertion that an ongoing, supportive professional development program can substantially improve classroom teachers’ physical education programs.

The purpose of the present study was to determine the extent to which intervention process measures are useful in predicting changes in cardiovascular disease (CVD) risk factors among subjects exposed to interventions. Subjects were 99 adult and 105 children who participated in an 18 session, family-based diet and exercise change program. During the intervention, each participant self-monitonally, diet and exercise for 12 weeks. Additinally, attendance, session evaluation, confidence to achieve goals and goal achievement data were collected each week. The intervention was successful in changing diet, blood pressure and cholesterol levels, but did not produce significant changes in exercise or body mass index. Stepwise multiple regression analyses were conducted for each outcome variable at the 1 and 2 year follow-ups. Ethnicity and sex were first forced into each regression. None of the process measures consistently predicted multiple outcomes in adults and children. The pattern of results provides limited support for the hypothesis that intervention process variables such as attendance, adherence to self–monitoring, achievement of goals and attitude toward sessions partially mediate intervention effects. It is concluded that process measures should be collected in health behavior change programs so that process-outcome relation ships can be further explored.

Neighborhood environment studies of physical activity (PA) have been mainly single-country focused. The International Prevalence Study (IPS) presented a rare opportunity to examine neighborhood features across countries. The purpose of this analysis was to: 1) detect international neighborhood typologies based on participants' response patterns to an environment survey and 2) to estimate associations between neighborhood environment patterns and PA.A Latent Class Analysis (LCA) was conducted on pooled IPS adults (N=11,541) aged 18 to 64 years old (mean=37.5±12.8 yrs; 55.6% women) from 11 countries including Belgium, Brazil, Canada, Colombia, Hong Kong, Japan, Lithuania, New Zealand, Norway, Sweden, and the U.S. This subset used the Physical Activity Neighborhood Environment Survey (PANES) that briefly assessed 7 attributes within 10-15 minutes walk of participants' residences, including residential density, access to shops/services, recreational facilities, public transit facilities, presence of sidewalks and bike paths, and personal safety. LCA derived meaningful subgroups from participants' response patterns to PANES items, and participants were assigned to neighborhood types. The validated short-form International Physical Activity Questionnaire (IPAQ) measured likelihood of meeting the 150 minutes/week PA guideline. To validate derived classes, meeting the guideline either by walking or total PA was regressed on neighborhood types using a weighted generalized linear regression model, adjusting for gender, age and country.A 5-subgroup solution fitted the dataset and was interpretable. Neighborhood types were labeled, "Overall Activity Supportive (52% of sample)", "High Walkable and Unsafe with Few Recreation Facilities (16%)", "Safe with Active Transport Facilities (12%)", "Transit and Shops Dense with Few Amenities (15%)", and "Safe but Activity Unsupportive (5%)". Country representation differed by type (e.g., U.S. disproportionally represented "Safe but Activity Unsupportive"). Compared to the Safe but Activity Unsupportive, two types showed greater odds of meeting PA guideline for walking outcome (High Walkable and Unsafe with Few Recreation Facilities, OR=2.26 (95% CI 1.18-4.31); Overall Activity Supportive, OR=1.90 (95% CI 1.13-3.21). Significant but smaller odds ratios were also found for total PA.Meaningful neighborhood patterns generalized across countries and explained practical differences in PA. These observational results support WHO/UN recommendations for programs and policies targeted to improve features of the neighborhood environment for PA.

73 This study evaluated the reliability and validity of the phone version of the 7-Day Physical Activity Recall (7-D PAR), a standardized interview. The in-person version of the 7-D PAR has been one of the most extensively studied self-report measures of physical activity and is shown to be valid and reliable. Efficiency in research would be improved if the 7-D PAR could be conducted via phone, but this version needs to be validated. Seventy-four adults (48 females, 26 males), ranging widely in age (18 to 67) and activity levels, were studied. Participants completed 7 days of activity monitoring using the Tritrac-R3D® (TT), an accelerometer which records movement in 3-dimensions. Participants completed activity questionnaires, and both versions of the 7-D PAR on separate days, in counter-balanced order. Reliability was assessed by comparing the phone and in-person 7-D PAR data for six matched days. Validity was examined by comparing 7-D PAR and TT data for 6 matched days. Intraclass correlations between the phone and in-person 7-D PAR total KKD (kcals*kg-1*day-1) were high (R=.957). Correlations of TT-assessed KKD were r=.413, p<.001 with the phone 7-D PAR and r=.445, p<.001 with the in-person 7-D PAR. Analyses of recall of minutes of physical activity at different intensity levels on the phone 7-D PAR and recorded by the TT indicate low correlations at “moderate” levels of activity (r=.153, p<.20), and higher correlations for“hard” (r=.508, p<.001) and “very hard”(r=.764, p<.001) activity. Correlations were similar for the in-person 7-D PAR (r=.112, p<.35; r=.665, p<.001; r=.663, p<.001 - “moderate”, “hard” and“very hard”, respectively). The phone and in-person 7-D PARs are highly reliable measures of physical activity. The validity of both the phone and in-person 7-D PARs, especially for reports of activity at higher intensity levels, is supported by the correlations with objective monitoring.

NF-kappaB is constitutively active in many solid tumors, including breast cancer. However, the role of NF-kappaB in breast carcinogenesis is unknown. Ikkalpha(AA/AA) "knockin" mice in which activation of IkappaB kinase alpha (IKKalpha) is prevented by replacement of activation loop serines with alanines exhibit delayed mammary gland growth during pregnancy, because IKKalpha activity is required for cyclin D1 induction and proliferation of lobuloalveolar epithelial cells. Given the role of cyclin D1 in breast and mammary cancer, we examined involvement of IKKalpha in mammary carcinogenesis induced by oncogenes or a chemical carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA). The Ikkalpha(AA) mutation retarded tumor development in response to either 7,12-dimethylbenzaanthracene or the MMTV-c-neu (ErbB2/Her2) transgene but had no effect on MMTV-v-Ha-ras-induced cancer, although both oncogenes rely on cyclin D1. Strikingly, primary Ikkalpha(AA/AA)/MMTV-c-neu carcinoma cells exhibited diminished self-renewal capacity, resulting in the inability to establish secondary tumors. Ikkalpha(AA/AA)/MMTV-c-neu carcinoma cells underwent premature senescence when cultured under conditions used for propagation of mammary gland stem cells. Thus, IKKalpha is not only a regulator of mammary epithelial proliferation, but is also an important contributor to ErbB2-induced oncogenesis, providing signals that maintain mammary tumor-initiating cells. IKKalpha may represent a novel and specific target for treatment of ErbB2-positive breast cancer.