Stroke is the most common cause of acquired epilepsy in persons 35 and older. The massive increase in extracellular glutamate during stroke causes a cascade of intracellular events that can lead to cell death or the molecular changes that initiate the development of epilepsy. In addition, many studies point to a modulatory role of the endocannabinoid system in controlling seizures. Animal models of stroke induced acquired epilepsy have been difficult to develop. Therefore, this dissertation was initiated to develop an organotypic hippocampal slice culture model of acquired epilepsy and examine the changes in distribution and function of the endogenous CB1 receptor system. We utilized 4-aminopyridine and glutamate to induce separate excitotoxic injuries to slice cultures. Both injuries produced significant cell death acutely following the injury. After a latency period, we observed a significant increase in the number of slice cultures that displayed electrographic seizures in both injury models. Western blot analysis demonstrated that the cannabinoid CB1 receptor protein was significantly upregulated following injury with glutamate. Immnohistochemical studies demonstrated that this receptor upregulation was likely specific to the glutamatergic terminals. Electrophysiological experiments were performed to study endocannabinoid modulation of inhibitory and excitatory signaling in the CA3 pyramidal cells. We demonstrated that depolarization induced suppression of excitation (DSE) was enhanced in slice cultures that had undergone glutamate injury. This indicated that the upregulation of CB1 receptors following glutamate injury was physiologically functional, as it enhanced cannabinoid control of the excitatory signaling. These studies support the hypothesis that there is a functional alteration of CB1 receptors in the epileptic state that acts to suppress seizures. The development of an organotypic hippocampal slice culture model of stroke acquired epilepsy provides a unique tool to study the neuronal plasticity changes associated with epileptogenesis. It also provides a practical model to study pharmacological agents that may be useful in preventing or treating epilepsy.

Sympathetic overactivity (SO) is associated with obesity and cardiovascular disease (CVD) mortality. Black adolescents have an increased risk of obesity and CVD later in life, particularly females. PURPOSE: To evaluate differences in SO between black (BOA) and white obese female adolescents (WOA). METHODS: Sixty-one BOA (n=49, 13.7±1.6 yrs, 38.1±6.1 kg/m2) and WOA (n=12, 13.3±2.2 yrs, 34.3±4.9 kg/m2) completed a maximal graded treadmill test after which an exercise recovery index (ERI; heart rate/VO2 plateau) was calculated. RESULTS: The ERI was significantly greater in BOA compared to WOA (29.8 ± 6.4 vs. 24.1 ± 3.1, P = 0.004). Multiple linear regression modeling revealed a significant independent association between ERI and VO2FFM (r = -0.310, P = 0.027) and %FAT (r = 0.326, P = 0.020) in BOA only. CONCLUSIONS: These results suggest that BOA females have greater SO than WOA females.

Most surgeries and many medical procedures commonly make use of some form of anesthesia to maximize patient comfort and safety. However, all are associated with risks. Obesity and related health care problems are relatively common in anesthesia and also have a negative effect on morbidity and mortality. Trends in pediatric obesity show increases in both the prevalence and risks for the development of other disease. Using the 1997 through 2009 Kids’ Inpatient Database (KID), this study will assess diagnostic codes to identify complications related to anesthesia in the obese pediatric population. Information gained from this study may serve to advance research and the development of anesthetic techniques to improve both safety and overall health for this population.

Background: Despite the numerous pharmaceutical applications of alcohols andglycols, the interspecies differences with respect to their pharmacokinetics (PK) arepoorly understood. The aim of this research was to use in-vivo and in-vitro approaches to compare and model the PK characteristics across various species.Methods: Appropriate published in-vivo studies (in different species) foralcohols and glycols were carefully selected. PK analysis was performed using (a) noncompartmental analysis and (b) compartmental modeling to estimate relevant dose-independent PK parameters. Next, six alcohols (methanol, ethanol, 1 -propanol, 1 -butanol, 1-hexanol, and 1-octanol), two glycols (ethylene glycol and propylene glycol)and one secondary alcohol (2-propanol) were examined as in-vitro substrates for equine ADH using a UV spectrophotometric assay to evaluate the effect of molecular structure. Furthermore, in-vitro metabolism of ethanol and propylene glycol was also characterized in hepatic cytosolic fractions from rat, rabbit, dog and human and in-vitro in-vivo correlation for the hepatic disposition parameters was assessed. Finally, allometric scaling relationships for ethanol and propylene glycol PK parameters (in-vivo and in-vitro) were developed and validated.Results: Alcohols and glycols exhibited nonlinear PK due to saturable hepaticmetabolism in all species. The reported in-vivo data were well described by oneltwo compartment PK models with parallel saturable metabolism and first-order renalexcretion. In-vitro equine ADH experiments revealed differences in affinity andturnover between the substrates: Enzyme affinity (1/Km) and in-vitro intrinsic clearance (CLintin-vitro) correlated positively with logP values; glycols showed lower CLintin-vitro values than straight-chain alcohols. In-vitro hepatic cytosol studies yielded acceptable in-vivo predictions for the metabolic clearance (CLmet) of ethanol and propylene glycol in the rat, dog and human, but not the rabbit. Vdss, Vmax, CLintin-vitro, CLmet scaled allometrically across species with similar powers for both ethanol and propylene glycol, and good agreement between in-vivo and in-vitro scaling was noted. The allometric scaling models gave excellent predictions when externally validated against in-vivo concentration-time data. Conclusions: The present research demonstrates .the successful application of amodeling-based approach to elucidate interspecies relationships for alcohols andglycols, compounds which exhibit nonlinear PK and mainly low hepatic extractionbehavior. The in-vitro experimental systems have been used successfully forcharacterizing alcohol/glycol metabolism and predicting in-vivo disposition.

Problem: There are a significant proportion of patients taking acetylcholinesterase inhibitors (ChEi) for cognitive dysfunction also taking medications with anticholinergic (ACh) properties that may counteract their effects. As the number of ACh medications, burden, increases so does the likelihood of an adverse outcome.Background: ACh medications are frequently used in the elderly population (Carnahan 2004) even those with dementia or AD (Roe et al., 2002; Giron et al., 2001; Altavela 2003; Gill et al., 2005; Kogut et al., 2005). Methods: Hospitalized patients > 65 years of age with dementia (AD, other dementias, or with inferred dementia based on ChEi or NMDA antagonist medication use) were studied using UHC clinical database. This document was created in Microsoft Word 2000. Results: Dementia patients on ChEi therapy were more likely to receive an ACh (chi-square 70.1, df=l, pConclusion: A person's age and mental health status along with their current drug regimen, such as ChEi therapy, need to be closely and carefully considered before deciding to use unnecessary ACh drugs in this population which can have detrimental effects.

Epilepsy, defined by recurrent seizures, is the one of the most prevalent neurological disorders worldwide (World Health Organization 2007). While many forms of epilepsy are well-controlled by anti-epileptogenic medications, a significant portion of patients have intractable, i.e. untreatable, seizures. The etiology of these seizures is varied, but a significant cause, particularly for patients with intractable epilepsy is developmental malformation. In these cases, an error or interruption during the development of the neocortex produces a structural alteration. Such patients may have other neurological problems, but seizures are the most common symptom. The neuronal mechanisms that link malformation and cortical hyperexcitability are not well understood. Here we have sought to examine potential mechanisms that result from microgyria, a malformation characterized by excessive numbers of small gyri.The presence of epileptiform activity indicates that the normal balance of excitation and inhibition has shifted . Two functions of inhibition within neocortex are to prevent spread of excitation, and to modulate the timing of surrounding excitation. Although seemingly contradictory, increasing some forms of inhibition can result in an increase in synchronous excitatory activity. We hypothesize that for certain malformation epilepsies, the inhibitory processes that control timing are increased, creating a hyper-synchronous cortex, while the inhibitory processes that control horizontal spread are decreased, allowing the propagation of such activity. Here we have examined the network effect of selectively modulating the inhibitory cells that control vertical or columnar cortical synchrony. This modulation is performed via activation of metabotropic glutamate receptors found on the vertically-projecting interneurons but not on those inhibitory cells that control horizontal spread of activity. Our results suggest that the network effect of activating these interneurons is altered in malformed, epileptogenic cortex.

The purpose of this prospective clinical study was to investigate differences in outcomes from two common treatment modalities used to reduce deep overbite: maxillary incisor intrusion using an intrusion arch and posterior tooth eruption using an anterior bite plate. Pre-treatment, post-overbite correction and post-treatment records were gathered from 32 patients who presented with deep overbite malocclusions to the Virginia Commonwealth University orthodontic clinic. Both groups of patients experienced reductions in overbite and maxillary incisor display as well as maxillary and mandibular incisor proclination and mandibular incisor occlusal movement during treatment. In the intrusion arch group, the center of resistance of the maxillary incisor was significantly intruded during overbite correction. The maxillary incisor incisal edge was significantly more intruded at the end of treatment in the intrusion arch group. Both groups experienced flattening of the smile arc in agreement with previous studies showing similar changes in orthodontically treated individuals.

Purpose: The purpose of this study is to examine the pain response experienced by children undergoing simple tooth extractions using 2% Lidocaine injection versus Oraqix topical anesthetic gel. Methods: This study is being conducted at VCU pediatric dentistry clinic. The sample size will consist of 15 children ages 7-12 undergoing a simple extraction procedure. Each participant is randomly assigned to one of two groups, the lidocaine injection group or the Oraqix topical group. The pain level will be measured at four key events during the procedure. The first event will evaluate the pain at baseline. The second event will measure pain during the anesthetic injection or Oraqix topical gel application. The third event will record the response after the tooth has been extracted and the final event will evaluate pain five minutes post operatively. The children are asked to rate their pain using the Facial Pain Scale after all four events. The dentist and an independent observer watching a video of the extraction will also examine and rate the pain responses of each child at each of the four events. Results: The first two participants received lidocaine injection and experienced some pain upon injection. This pain was supported by what the dentist rated as well. One child felt pain on extraction and the other felt nothing. The dentist rated both children as feeling pain. The Oraqix child felt nothing upon application but felt pain during the extraction and post-operatively. The dentist rated the child as feeling nothing during the entire procedure. Conclusions: It appears that the lidocaine injection group’s pain rating matches the pain rating given by the dentist. The Oraqix patient experienced no pain upon application, but did feel pain upon extraction and five minutes post-op. The dentist’s rating contradicted this by rating the child as feeling no pain through the entire procedure. This study is limited by the number of participants and needs more patients to further evaluate other children’s pain responses.

AIM: This study aims to determine what portion of specific autoantibody phenotypes are genetically determined by using a twin model. METHODS: This study specifically examines Anti-Ro(SSA), Anti-La (SSB), Anti-Sn/RNP, Anti-Sm, Anti-Jo-1, Anti-Scl-70, Anti-Tg & Anti-TPO, Anti-dsDNA, Anti-PS, and Anti-cardiolipin antibodies for their heritability. This study examined 104 same-sex adult twins (66 monozygous, 38 dizygous) for the above mentioned autoantibody values. The serum autoantibody values in each subject were quantified using automated ELISA. Descriptive statistics including, distributions, quantiles, and moments were calculated by zygosity for continuous antibody values, subject ages, gender, race and smoking status. Categorical antibody levels were used to determine twin pair concordance rates. Continuous and rank ordered autoantibody values were used to determine the presence and portion of a genetic component. To evaluate how strongly the antibody values in each twin group resembled each other, the intraclass correlation was calculated for each antibody by zygosity. The genetic variances, environmental variances, and heritability were estimated using path models with maximum likelihood estimation techniques. The phenotypic variance was modeled as a linear function of underlying additive genetic (A), dominant genetic (D), common environmental (C), and random environmental (E) effects. RESULTS: Several antibodies demonstrated a genetic component in our study population. Anti-cardiolipin had a genetic component with an estimated 69% heritability. Anti-dsDNA yielded a genetic component with a heritability estimate of 55-62%. Anti-Jo-1 presented a genetic component with the heritability estimate to be 41-51%. Anti-SCL-70 demonstrated a genetic component with a heritability estimate of 42-59%. Anti-PL had a genetic component with a heritability estimate of 52-54%. Several antibodies did not have a measurable genetic component. These included anti-Sm, anti-Ro(SSA), anti-La(SSB), anti-sn/RNP, anti-Tg, and anti-TPO. Some possibilities for the lack of a measureable genetic component may be due to the limited number of discordant twin pairs and/or the small number of subjects with higher levels of antibodies. CONCLUSION: The results of this study suggest several clinically relevant markers of auto-immunity may be partially genetically determined. These include: anti-cardiolipin, anti-dsDNA, anti-Jo-1, anti-SCL-70, and anti-phospholid.

Functional MRI technique is vital in investigating the effect of an intervention on cortical activation in normal and patient population. In many such investigations, block stimulation paradigms are still the preferred method of inducing brain activation during functional imaging sessions because of the high BOLD response, ease in implementation and subject compliance especially in patient population. However, effect of an intervention can be validly interpreted only after reproducibility of a detectable BOLD response evoked by the stimulation paradigm is first verified in the absence of the intervention. Detecting a large BOLD response that is also reproducible is a difficult task particularly in olfactory Functional MRI studies due to the factors such as (a) susceptibility-induced signal loss in olfactory related brain areas and (b) desensitization to odors due to prolonged odor stimulation, which is typical when block paradigms are used. Therefore, when block paradigms are used in olfactory interventional Functional MRI studies, the effect of the intervention may not be easily interpretable due to the factors mentioned above. The first task of this thesis was to select a block stimulation paradigm that would produce a large and reproducible BOLD response. It was hypothesized that a BOLD response of this nature could be produced if within-block and across-session desensitization could be minimized and further, that desensitization could be minimized by reducing the amount of odor by pulsing the odor stimulus within a block instead of providing a continuous odor throughout the block duration. Once the best paradigm was selected, the second task of the thesis was to select the best model for use in general linear model (GLM) analysis of the functional data, so that robust activation is detected in olfactory related brain areas. Finally, the third task was to apply the paradigm and model that were selected as the best among the ones tested in this thesis, to an olfactory interventional Functional MRI study investigating the effect of food (bananas) eaten to satiety on the brain activation to the odor related to that food. The methods used in this thesis to ensure valid interpretation of an interventional effect, can serve as a template for the experimental design of future interventional Functional MRI studies.