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Background and objectivePediatricians are encouraged to engage in community child health activities, yet practice constraints and personal factors may limit involvement. The objective was to compare community involvement in 2004 and 2010 and factors associated with participation in the past year.MethodsAnalysis of 2 national mailed surveys of pediatricians (2004: n = 881; response rate of 58%; 2010: n = 820; response rate of 60%). Respondents reported personal characteristics (age, gender, marital status, child ≤5 years old, underrepresented in medicine), practice characteristics (type, setting, full-time status, time spent in general pediatrics), formal community pediatrics training, and community pediatrics involvement and related perspectives. We used χ2 statistics to measure associations of personal and practice characteristics, previous training, and perspectives with involvement in the past 12 months. Logistic regression assessed independent contributions.ResultsFewer pediatricians were involved in community child health in 2010 (45.1% in 2004 vs. 39.9% in 2010) with a higher percentage participating as volunteers (79.5% vs. 85.8%; both P = .03). In 2010, fewer reported formal training at any time (56.1% vs. 42.9%), although more reported training specifically in residency (22.0% vs. 28.4%; both P < .05). Factors associated with participation in 2010 included older age, not having children ≤5 years old, practice in rural settings, practice type, training, and feeling moderately/very responsible for child health. In adjusted models, older age, practice setting and type, feeling responsible, and training were associated with involvement (P < .05).ConclusionsFormal training is associated with community child health involvement. Efforts are needed to understand how content, delivery, and timing of training influence involvement.

Since the completion of human genome sequencing, scientists have attempted to decipher the interactions and distribution of functional elements in the genome in hopes of explaining the mechanisms behind cancer development. Recently, researchers have been studying epigenomics, as the reversible nature of epigenomic alterations has greatly aided the understanding of the underlying mechanisms in tumour development. The major epigenetic events include DNA methylation and histone modifications. The first part of my thesis focuses on studying the interactions among DNA methylation and various histone H3K9 modifications. At any given locus, there are 243 possible patterns of these epigenetic marks. Through global epigenetic mapping, I identified non-random distribution of these patterns and discovered that some specific patterns are more prominent than others. The fact that these histone marks are not distributed equally at large scale suggests that histone modification is not only a small scale phenomenon. To extend studies of modification on DNA, the second part of my thesis focuses on studying cisplatin induced DNA adducts and their clearance. I achieved this by adapting the immunoprecipitation technique that was optimized in the first part of my thesis. My results showed non-random integration of cisplatin on DNA. Furthermore, from global clearance maps, I identified slow drug clearance and rapid drug clearance loci. This implies that there are some adduct formation sites that are being preferentially repaired over others. With more interest on the interaction between chromatin structure and DNA damage, the global maps generated in my thesis provide a critical step in understanding the distribution of these events in the genome, and will contribute to establishing a biological model investigating effects of cisplatin induced alterations on chromatin modification structure.

It has been questioned whether there are receptors for urokinase-type plasminogen activator (uPA) that facilitate plasminogen activation other than the high affinity uPA receptor (uPAR/CD87) since studies of uPAR knockout mice did not support a major role of uPAR in plasminogen activation. uPA also promotes cell adhesion, chemotaxis, and proliferation besides plasminogen activation. These uPA-induced signaling events are not mediated by uPAR, but mediated by unidentified, lower-affinity receptors for the uPA kringle. We found that uPA binds specifically to integrin alpha v beta 3 on CHO cells depleted of uPAR. The binding of uPA to alpha v beta 3 required the uPA kringle domain. The isolated uPA kringle domain binds specifically to purified, recombinant soluble, and cell surface alpha v beta 3, and other integrins (alpha 4 beta 1 and alpha 9 beta 1), and induced migration of CHO cells in an alpha v beta 3-dependent manner. The binding of the uPA kringle to alpha v beta 3 and uPA kringle-induced alpha v beta 3-dependent cell migration were blocked by homologous plasminogen kringles 1-3 or 1-4 (angiostatin), a known integrin antagonist. We studied whether the binding of uPA to integrin alpha v beta 3 through the kringle domain plays a role in plasminogen activation. On CHO cell depleted of uPAR, uPA enhanced plasminogen activation in a kringle and alpha v beta 3-depenclent manner. Endothelial cells bound to and migrated on uPA and uPA kringle in an alpha v beta 3-dependent manner. These results suggest that uPA binding to integrins through the kringle domain plays an important role in both plasminogen activation and uPA-induced intracellular signaling. The uPA kringle-integrin interaction may represent a novel therapeutic target for cancer, inflammation, and vascular remodeling.

Autonomic nervous system (ANS) activity is related to body weight regulation. Based on the hypothesis that Most Obesities kNown Are Low In Sympathetic Activity (MONA LISA), it has been suggested that most obese subjects and animals have low sympathetic nervous system activity. Leptin, leptin receptor, and ghrelin genes influence the ANS regulation of body weight and food intake. The aim of this study was to investigate whether there are differences in leptin, the leptin receptor, or ghrelin regulation between lines of chickens selected for high (HWS) or low body weight (LWS). Intraperitoneal injections of reserpine were administrated to chickens from the HWS and LWS lines. Body weight and food intake were then compared to evaluate ANS regulation. While reserpine caused a transitory decrease in food intake and body weight in both lines, the magnitude of the change was greater in the HWS than in the LWS chickens. However, chickens from the LWS line exhibited greater catecholamine and indoleamine level changes in response to reserpine than those from the HWS line. Therefore, HWS chickens were more sensitive to the body weight-reducing effects of reserpine than LWS lines, while LWS chickens appeared to have greater sympathetic nervous system activity. Food and water intakes were differentially affected in HWS and LWS chickens in response to intracerebroventricular administration of human recombinant leptin. Leptin caused a linear decrease in food intake in the LWS line, but no effect on food intake in the HWS lines. The HWS chickens tended to have reduced water intake following leptin administration. These results suggest that the leptin receptor, or the down-stream neuropeptide regulation pathway mediating the effect of leptin; may be different between chickens from the HWS and LWS lines. Leptin, insulin like growth factor (IGF)-1, and IGF-2 concentrations in the plasma of HWS and LWS lines of chickens were evaluated. Leptin, IGF-1 and IGF-2 levels were significantly higher in the LWS than HWS chickens. The HWS female leptin concentrations were significantly lower than in HWS males or LWS females. Male chickens had greater IGF-1 concentrations in the plasma than female chickens. However, the concentration of IGF-2 did not differ between sexes. The difference in leptin concentrations in these lines and sexes may explain the differences in age of sexual maturity. Different IGF-1 and IGF-2 concentrations may be involved in the obese and anorexic conditions, fast and slow growth, and high and low food consumption found in these two lines of chickens. Differences in the gene sequence of the leptin receptor were observed in HWS and LWS lines of chickens. A single nucleotide polymorphism (SNP) in the intron between exon 8 and 9 introduced a restriction site for the enzyme Sel I in the HWS, but not the LWS line. Two SNP were detected in the leptin receptor cDNA region at nucleotides 189 and 234. At nucleotide 189, the LWS line has both a homozygous (T-T) and heterozygous (C-T), whereas the HWS line has only homozygous (T-T) form. The SNP found in nucleotide 234 introduces a restriction site Mse I in the HWS, but not the LWS line. These specific changes may be directly involved or closely linked to differences between the two lines in either the coding or regulatory domains of the leptin receptor. Differences in the leptin receptor gene expression between HWS and LWS lines of chickens in various organs and ages were observed. Leptin receptor expression in the whole brain was significantly different between sexes at 28 days-of-age in the HWS and LWS lines. The LWS line had higher leptin receptor gene expression in the liver at 2 days-of-age than at 56 and 363 days-of-age, but no differences were observed in the HWS line. In addition, at 2 days-of age, liver leptin receptor gene expression was higher in LWS than HWS chickens, but the reverse was observed at 363 days-of age. In adipose tissue, leptin receptor expression was higher in the LWS than HWS line. Leptin receptor expression in adipose tissue was greater at 363, than 28 and 56 days-of-ages. Our results showed that changes in the regulation of leptin and the leptin receptor were associated with sex, age, and growth. Differences in the ghrelin gene in the HWS and LWS lines under different feeding conditions were investigated. Both HWS and LWS chickens have six extra base pairs in the 5'-untranslated region. The LWS male ghrelin gene expression was significantly lower than in the LWS female and HWS male. The 84 day-old males had lower gene expression than 84 day-old females and 363 day-old males. When comparing different feeding methods, females allowed ad libitum feed consumption had a lower cycle threshold cycle number (CT) ratio than males allowed ad libitum feeding or fasted females. However, the inflection point cycle number of ad libitum fed females was lower than that of the ad libitum fed males, but greater than the fasted females. Ghrelin gene expression was different between the two lines of chickens, and the expression of ghrelin in chickens was influenced by body weight selection, sex, age, and feeding condition. Ph. D.

Autonomic nervous system activity is related to body weight regulation. Based on the MONA LISA hypothesis it has been suggested that most obese subjects and animals have low sympathetic nervous system activity. The aim of this study was to investigate whether there are differences in autonomic nervous system activity between lines of chickens selected for either high (HWS) or low body weight (LWS). In Exp. 1, various pharmacological agents were injected intravenously, and the changes in blood pressure (BP) and heart rate (HR) of both HWS and LWS chickens were compared. The results showed that the HWS birds had a greater increase in BP and HR than the LWS following injection of atropine, a muscarinic receptor blocker, and LWS birds had a greater decrease in BP and HR to propranolol, a beta- adrenergic receptor blocker than the HWS birds. These results suggested that HWS chickens have higher parasympathetic tone, whereas LWS chickens have a higher sympathetic nervous system tone regulating the cardiovascular system. HWS and LWS chickens displayed a similar response in BP and HR following injection of the ganglion blocker tetraethylammonium chloride. These results suggest that there is no significant difference in the central autonomic nervous system in the cardiovascular regulation between HWS and LWS together. Since there does not appear to be any differences in the activity of the autonomic nervous system activity at the level of the central nervous system, these findings imply that the difference in response to atropine and propranolol could be caused by differences in adrenal activity. The ratio of heart rate and blood pressure after the injection of phenylephrine showed significant difference between these two lines of birds, but not when phenylephrine was injected following atropine. This result indicated that HWS are more dependent on the parasympathetic nervous system to regulate the baroreceptor reflex. The percentage of adrenal and sympathetic impact on the regulation of heart rate showed that LWS females required greater adrenal activity than the other birds. In Exp. 2, the body weight and food intake responses of HWS and LWS chickens to ip injections of reserpine were compared. Reserpine caused a transitory decrease in food intake and body weight in both lines of birds. However HWS chickens recovered more slowly from the depression caused by reserpine than the LWS chickens. This could be due to lower sympathetic nervous system activity. In conclusion, it appears that HWS may have lower sympathetic activity than LWS. Combining the results of both experiments, it appears that the HWS birds have lower sympathetic and higher parasympathetic activity. Furthermore central nervous system autonomic activity in BP and HR regulation is not different between HWS and LWS, but the activity of the adrenal gland may be different between these two lines of birds. Master of Science