Endothelial dysfunction, or the inability of an artery to dilate sufficiently when subjected to excessive shear stress, serves both as a predictor of future cardiovascular events as well as an early indication of atherosclerosis. Several chronic disease states, including obesity, have been shown to alter endothelial function, which may be mediated through circulating pro- and anti-inflammatory adipokines. Still, the mechanisms by which obesity-related low-grade inflammation alters endothelial function are not fully elucidated. Acute and chronic endurance exercise training has previously been shown to be effective in improving endothelial function; however, chronic resistance exercise training is not universally regarded as beneficial to vascular functioning. Far fewer studies have examined the effect of acute resistance exercise on vascular function and adipokine release. To further understand the effects of resistance exercise training on vascular function, a meta-analysis was completed to examine the effects of resistance training on brachial artery flow mediated dilation (FMD), a common measure of endothelial function. The results of the meta-analysis indicate that resistance training has a small positive effect on FMD. Additionally, the effects of an acute bout of lower body resistance exercise on forearm blood flow (FBF) and two inflammatory cytokines were evaluated in obese (>30% body fat) and non-obese (≤30% body fat) subjects. It was hypothesized that the resistance exercise bout would increase FBF, that those changes would be greater in obese versus non-obese subjects, and that the changes in circulating cytokines (adiponectin and tumor necrosis factor-α) would be related to changes in FBF. The results indicate that FBF measures in obese and non-obese subjects react in a divergent pattern immediately following resistance exercise but return to baseline within 24 hours. These changes were not related to changes in adiponectin or TNF-α although changes in adiponectin were related to changes in TNF-α. In conclusion, resistance exercise training programs may have a small positive effect on vascular function which may reduce overall cardiovascular disease risk. Additionally, obese and non-obese subjects display differing patterns of vascular responses to an acute bout of resistance exercise, supporting the view that obesity, and its associated low-grade inflammatory response, may negatively alter vascular homeostasis.

The human Class Ib gene, HLA-E inhibits both Natural Killer Cells and a subset of CD8+ cytotoxic T lymphocytes by engaging the CD94/NKG2A inhibitory receptor. IFN-γ induces the expression of HLA-E as well as Class Ia molecules, which are required for the killing of target cells. Since HLA-E has negative effects on immune killing of target cells, we have sought to identify locus specific mechanisms of IFN-γ induction in order to identify molecular targets for selective activation of Class Ia genes, but not HLA-E. We have previously identified a unique upstream IFN-γ response region in the HLA-E promoter and showed that GATA-1 is required for its function in the K562 leukemic cell line. We have now examined the effect of GATA family members on IFN-γ induction of HLA-E in other cell types. HLA-E CAT reporter gene assays demonstrate that tumor cells that express GATA factors as determined by western blot and quantitative PCR, mediate a 2.4 to 4.0 fold enhanced response to IFN-γ stimulation. Functional constructs containing mutations of the core nucleotides in the GATA binding site had a 4.8 fold decreased response to IFN-γ in A2780 cells and a 8.5 to 14.0 fold decreased response to IFN-γ in SKOV3 cells. Knockdown of GATA-6 using siRNA resulted in a 40% decrease in HLA-E induction in Seg1 cells and a 30% decrease in HLA-E induction in HCT116 cells. Tetracycline regulated shRNA knockdown of GATA-6 expression in the SKOV3 cell line revealed a 3 fold decrease in the IFN-γ response of HLA-E reporter driven constructs. Additionally we observed a decreased IFN-γ response in SKOV3 cells transfected with siRNA specific for CBP and IRF-9. We conclude that GATA factors play a tissue specific role in regulation of IFN-γ mediated HLA-E expression and that IRF-9 may be a target for the differential manipulation of classical MHC and HLA-E.

ANESTHESIA RECORDKEEPING: ACCURACY OF RECALL WITH COMPUTERIZED AND MANUAL ENTRY RECORDKEEPING By Thomas Corey Davis, PhD A dissertation submitted in partial fulfillment of the requirements for the degree of PhD in Health Related Sciences at Virginia Commonwealth University. Virginia Commonwealth University, 2011 Major Director: Dr. Chuck Biddle Director of Research, Department of Nurse Anesthesia And Dr. Jeffery A. Green Assistant Chief of Anesthesiology, Department of Anesthesia Introduction: Anesthesia information management systems are rapidly gaining widespread acceptance. Aggressively promoted as an improvement to manual-entry recordkeeping systems in the areas of accuracy, quality improvement, billing and vigilance, these systems record all patient vital signs and parameters, providing a legible hard copy and permanent electronic record. At risk is a potential loss of “connectedness” to the patient with the use of computerized recordkeeping, perhaps jeopardizing vigilance. Methods: This research analyzed differences in the accuracy of Certified Registered Nurse Anesthetists' (CRNAs) recall of specific patient variables during the course of an actual anesthetic case. CRNAs using computerized recordkeeping systems were compared to CRNAs using manual entry recordkeeping. Accuracy of recalled values of 10 patient variables was measured - highest and lowest heart rate, systolic blood pressure, inspiratory pressure, and end-tidal carbon dioxide levels, lowest oxygen saturation and total fluid volume. In addition, a filmed educational vignette was presented to evaluate any effect on accuracy of recall following this presentation. Four tertiary care facilities participated in this research. A Solomon four-group research design was selected to control for the effect of pretesting on results of the filmed educational treatment. Results: 214 subjects participated in this study; 106 in the computerized recordkeeping group, and 108 in the manual entry recordkeeping group. Demographic covariates were analyzed to ensure homogeneity between groups and facilities. No significant statistical differences were identified between the accuracy of recall among the groups. There was no statistically significant effect of the educational film vignette on accuracy of recall. Conclusions: There was no difference in the accuracy of practitioners’ recall of patient variables when using computerized or manual entry recordkeeping systems, suggesting little impact on vigilance. The educational film presented did not have an effect on accuracy of recall following the discussion of benefits and limitations of methods of recordkeeping.

CD23, the natural negative regulator of IgE, has been shown to be involved in asthma progression through its regulation of IgE. To investigate if its sheddase, ADAM10, is also involved in asthma progression, three mouse models were utilized; an IgE/mast cell dependent model, an IgE dependent, mast cell independent model and a mast cell and IgE independent model. Experimental asthma was then induced in mice which were selectively deficient for ADAM10 in B cells (ADAM10-/-) and compared to WT controls. The ADAM-/- mice had decreased signs of asthma, including eosinophilia, AHR and IgE synthesis in the IgE dependent model compared to LM controls, while with the IgE independent model there was no significant difference. Thus, CD23Tg and ADAM10-/- B cell mice have reduced IgE dependent lung inflammation in mouse models compared to WT controls. As a follow up, ADAM10 was inhibited in WT mice by intranasal administration of an ADAM10 inhibitor, compared to carrier (DMSO) treated mice. As with ADAM10-/- mice, inhibition of ADAM10 was only able to control IgE dependent models. These results thus show that ADAM10 is a possible target in controlling IgE dependent allergic disease, possibly as blocking ADAM10 would cause an increase in CD23 membrane expression. To better understand how ADAM10 cleaves CD23 we first sought to confirm previous studies that CD23 is internalized, with the hypothesis that shedding takes place intracellularly, rather than at the cell surface as previously assumed. Indeed, ADAM10 is more highly expressed intracellularly than at the cell surface. At 37 ºC, crosslinking CD23, especially with the anti-stalk mAb 19G5, resulted in extensive CD23 internalization. In addition, the expected increase in soluble CD23 (sCD23) production when 19G5 was added was blocked by the addition of NH4Cl. NH4Cl is known to block the progression of the endosomal pathway. These findings thus confirmed our hypothesis that cleavage of CD23 requires internalization and progression through the endosomal pathway before it is released into the extracellular space. We further demonstrated that ADAM10 is not only involved in cleaving CD23, but also in sorting CD23 into exosomes, as B cells lacking ADAM10 do not incorporate CD23 into exosomes. In addition, we found that exosomes secreted from the cell contain full length CD23, thus showing that they could bind IgE/antigen complex and be involved in the known CD23 dependent enhancement of antigen presentation by the injection of IgE/antigen complexes compared to antigen alone. These results also show that the change in ADAM10 expression specifically in a B cell could be involved in enhancement of IgE dependent inflammation. To determine what signals change ADAM10 expression, ADAM10 promoter studies were initiated. We found that both IL-21 and anti-CD40 increased ADAM10 promoter activity, while IL-4 and IL-13 had no effect. Overall our data show that increasing ADAM10 activity and expression leads to increased inflammation and IgE and is a possible target in controlling IgE dependent diseases.

PURPOSE: To create a study designed to assess patient satisfaction and preference for oral versus intravenous sedation in conjunction with periodontal surgical procedures. METHODS: Twenty-six patients who required at least two periodontal surgery procedures and requested sedation for treatment, participated in our study at VCU Department of Periodontics. This was a randomized, cross-over design with groups which received an intravenous sedative regimen with or without oral sedation premedication for one surgery and oral sedation medication alone for the other surgery. The primary outcome measurement was the type of sedation preferred by the subject. RESULTS: 14/26 (53.8%) subjects indicated a preference for intravenous sedation, compared with 7/26 (26.9%) subjects who preferred oral sedation alone. 1/26 (3.8%) subject reported that they would prefer no sedation after experiencing both oral and oral/intravenous combination sedation methods. 4/26 (15.3%) of the subjects who completed the study reported “No Difference” with regards to their preference for either method of sedation. CONCLUSION: More subjects preferred intravenous sedation and would consent to the sedation again for any future needed surgery. This study supports the need to offer intravenous sedation with periodontal surgery

Purpose: The specific aim of this study is to evaluate the trends in dental health care for individuals with ectodermal dysplasia. Methods: This was a cross sectional analysis of subjects recruited through the National Foundation of Ectodermal Dysplasia (NFED). From 1997 to 2000, individuals with ectodermal dysplasia or their caregiver (if the individuals were too young to selfreport) voluntarily completed questionnaires. The questionnaire consisted of 37 items consisting of demographics, ectodermal dysplasia diagnosis, access to dental care, level of dental utilization, and type of dental services received. Descriptive statistics were used in addition to ANOVA analyses to evaluate the changing trends in oral health care for individuals with ectodermal dysplasia. Results: Preliminary results indicate: 1) individuals with ectodermal dysplasia are being diagnosed earlier than in the past, 2) physicians are primary source of the initial diagnosis of ectodermal dysplasia, 3) children with ectodermal dysplasia are receiving prostheses earlier than in the past, and 4) access to care is problematic. Conclusion: Diagnosis and recognition of treatment needs are occurring at an earlier age and that an access to dental care for individuals with ectodermal dysplasia continues to be an issue.

Dendritic spines are the major site of excitatory synapses in cortex, and factors that reduce dendritic spine numbers will produce serious cortical processing deficits, such as has been demonstrated for mental retardation and other psychiatric disorders. Prenatal alcohol exposure also has detrimental effects on brain development that lead to Fetal Alcohol Spectrum Disorder (FASD), which results in reduction of dendritic spine numbers in the hippocampus, prefrontal cortex and somatosensory cortex. FASD also is associated with temporal processing disorders involving sequential auditory stimuli that would be processed in auditory cortical areas. However, it is unknown if the reduction of spine density following prenatal alcohol exposure occurs at auditory cortex, or is generally reduced across the different sensory cortices. This present study examined that question. Young adult ferrets (176 days old, 1 male, 1 female), that were exposed to alcohol during the equivalent of third-trimester development, were used to prepare Golgi-Cox stained sections through primary auditory cortex (A1). Other cortical regions examined included primary somatosensory (S1), and higher-level multisensory cortices of lateral rostral suprasylvian (LRSS) and rostral posterior parietal (PPr) areas. Control values from normal animals (n=3) were derived from a previous study. The results of this present study demonstrated that, dendritic spine density was significantly (Student's t-test, P < 0.05) lower in the alcohol treated group than in normal controls in all the cortical regions examined. These data indicate that although reduced spine density in auditory cortex may underlie temporal processing disorders in FASD, pre-natal alcohol exposure has widespread consequences for sensory cortical processing in general.

Borrelia hermsii, an etiologic agent of tick-borne relapsing fever, binds negative complement regulator factor H (FH) via its FhbA protein. Direct demonstration of the role of FhbA in the disease process has been hindered by the lack of genetic manipulation systems for the relapsing fever Borrelia. Here, we demonstrate successful generation of a B. hermsii strain YOR fhbA deletion mutant (Bh YORΔfhbA) that constitutively produces green fluorescent protein (GFP). Genetic manipulation did not affect growth rate or plasmid composition. Bh YORΔfhbA lost factor H-binding and C3b-inactivation capabilities, but retained resistance to killing in human serum and infectivity in mice. Stable production of GFP was demonstrated in vitro and in vivo. Collectively, these results suggest that B. hermsii employs an unidentified mechanism of complement evasion that is FH-independent and sufficient for persistence within the host. Additionally, this study represents a significant methodological advancement in the molecular characterization of relapsing fever spirochetes.

The purpose of this in-vitro study is to compare the accuracy and reliability of a 3rd and 4th generation electronic apex locator (EAL) in locating the apical foramen when using insulated and non-insulated K files. Forty extracted human adult single-rooted teeth were coronally sectioned and placed in agar. EAL determined tooth length measurements were compared to actual tooth measurements. Comparisons to the standard measures used correlation and paired t-test. Preliminary comparisons of the groups used ANOVA to compare the means and the Brown-Forsythe test to compare variance. In the final analyses, the measurements were compared using a repeated-measures mixed-model multiway ANOVA that allowed for heterogeneous variance in the subgroups. Findings were that accuracy is not different due to insulation in the Root ZX group (p-value=0.50) but is improved in the Elements Diagnostic Unit group (p-value<.001). Reliability is nominally improved with insulation in both the Root ZX and Elements Diagnostic Unit.

Etiological models of complex disease are elusive[46, 33, 9], as are consistently replicable findings for major genetic susceptibility loci[54, 14, 15, 24]. Commonly-cited explanations invoke low-frequency genomic variation[41], allelic heterogeneity at susceptibility loci[33, 30], variable etiological trajectories[18, 17], and epistatic effects between multiple loci; these represent among the most methodologically-challenging issues in molecular genetic studies of complex traits. The response has been con- sistently reactionary—hypotheses regarding the relative contributions of known func- tional elements, or emphasizing a greater role of rare variation[46, 33] have undergone periodic revision, driving increasingly collaborative efforts to ascertain greater numbers of participants and which assay a rapidly-expanding catalogue of human genetic variation. Major deep-sequencing initiatives, such as the 1,000 Genomes Project, are currently identifying human polymorphic sites at frequencies previously unassailable and, not ten years after publication of the first major genome-wide association find- ings, re-sequencing has already begun to displace GWAS as the standard for genetic analysis of complex traits. With studies of complex disease primed for an unprecedented survey of human genetic variation, it is essential that human geneticists address several prominent, problematic aspects of this research. Realizations regarding the boundaries of human traits previously considered to be effectively disparate in presentation[44, 39, 35, 27, 25, 12, 4, 13], as well as profound insight into the extent of human genetic diversity[23, 22] are not without consequence. Whereas the resolution of fine-mapping studies have undergone persistent refinement, recent polygenic findings suggest a less discriminant basis of genetic liability, raising the question of what a given, unitary association finding actually represents. Furthermore, realistic expectations regarding the pattern of findings for a particular genetic factor between or even within populations remain unclear. Of interest herein are methodologies which exploit the finite extent of genomic variability within human populations to distinguish single-point and cumulative group differences in liability to complex traits, the range of allele frequencies for which common association tests are appropriate, and the relevant dimensionality of common genetic variation within ethnically-concordant but differentially ascertained populations. Using high-density SNP genotype data, we consider both hypothesis-driven and agnostic (genome-wide) approaches to association analysis, and address specific issues pertaining to empirical significance and the statistical properties of commonly- applied tests. Lastly, we demonstrate a novel perspective of genome-wide genetic “background” through exhaustive evaluation of fundamental, stochastic genetic processes in a sample of matched affected and unaffected siblings selected from high- density schizophrenia families.