This study was an effort to objectify and test empirically Tillich's theory. Its purpose was to investigate certain traditional aspects of religion to see whether their presence was associated with purposefulness in life. The variables used in this study were measures of participation in a faith, belief in an afterlife, belief in God, past and current doubts, religious experiences, commitment implied in the consideration of a church related vocation, and of purpose in life. Tillich would say that participation in a faith, a belief in God, and a commitment of some kind are essential elements of one's ultimate concern. The belief in an after life is a common way of avoiding the anxiety of death, which is an element of the anxiety of meaninglessness of today. Doubt concerning the Ultimate undermines ones meaning or purpose in life.

Porphyromonas gingivalis, a gram-negative, anaerobic bacterium, is widely recognized as a causative agent for periodontal disease. Despite sequencing of the complete genome, no research exists examining gene regulation response in P. gingivalis to shifts in pH. Previous studies have shown that P. gingivalis is capable of surviving in the variety of micro- environmental niches found within the oral cavity, including basic and acidic pH conditions. However, the underlying mechanisms of this survival are not well understood. This study examined P. gingivalis by comparing bacteria shocked at three acidic to neutral pH conditions (5.5, 6.5 and 7.0) to bacteria vii shocked at pH 8.5. Using microarray to examine global gene expression, differential gene expression was identified in all conditions, with total genes differentially regulated ranging from 30 to almost 500 genes. Among these, genes for ammonia production, and cation gradients were found significantly up-regulated in acidic conditions, indicating a possible role in base creation and cation transport for survival of P. gingivalis in adverse pH conditions.

Ramphogordius sanguineus (Rathke 1799) is a gregarious nemertean with a worldwide distribution and found mainly on hard substrates associated with mussels, oysters and other organisms of the fouling community. Asexual reproduction occurs by spontaneous fragmentation and only anecdotal accounts of sexual reproduction exist. This is the first phylogeographic study of R. sanguineus as well as the first species delimitation analyses employing DNA markers. Analysis of the mitochondrial gene nad6 and nuclear ISSR markers showed little diversity among geographically widespread populations, but AMOVA analyses of both markers revealed moderate to high genetic differentiation. Populations from Maine and Massachusetts exhibited the highest level of differentiation. These findings are consistent with predictions for invertebrates lacking a planktonic larval stage. Results of the nad6 tree-based delimitation analysis were in agreement with modern morphological and histocompatibility observations, suggesting that R. sanguineus is a single species and that a former division into four separate species was solely based on geographic location.

The thesis contains two manuscripts- 1) Developmental changes in pectoral muscle fiber diameter and number in the Blue Catfish, Ictalurus furcatus and 2) Reduction in pectoral spine and girdle in domesticated Channel Catfish, Ictalurus punctatus is likely caused by the absence of fish predators. The first study showed that fiber diameter increased linearly with fish size, whereas fiber number increases non linearly with fish size in the Blue Catfish correlated with dietary shift. The second study showed shorter spines in domesticated Channel Catfish as compared to wild Channel Catfish was a result of reduction in selection pressure during domestication in the absence of fish predators.

The contractility of smooth muscle in inflammatory bowel disease and experimental colitis is reduced due to inhibition of neurotransmitter release and a decrease in the response of smooth muscle to contractile agonists. We and others have shown that inflammation induced by TNBS treatment alters the expression and/or activity of signaling molecules involved in the regulation of Ca2+ mobilization, MLC20 phosphorylation and contraction in colonic smooth muscle. Although, thin filament- associated proteins such as calponin, caldesmon, tropomyosin and smoothelin do not directly participate in contraction, they regulate acto-myosin interaction and thus, muscle contraction. Calponin, caldesmon and tropomyosin inhibit actomyosin interaction and the inhibition is relieved upon phosphorylation of these proteins. Recent studies have shown that visceral smooth muscle from smoothelin knockout mice exhibited decreased contraction. However, the effect of inflammation on the expression of thin filament- associated proteins is not known. The aim of the present study is to determine the changes in the expression of calponin, caldesmon, tropomyosin, and smoothelin in colonic circular smooth muscle from TNBS- and DSS-induced colitis in mice. The animals were euthanized on day 3 and a segment of inflamed distal colon was removed. Colonic muscle strips from colitis mice and control mice were dissected for western blot and real-time RT-PCR analysis; contraction was measured by scanning micrometry in cells isolated from the muscle strips. Contraction in response to acetylcholine in muscle cells isolated from colonic muscle strips derived from mice with TNBS colitis was significantly inhibited compared with the response of cells derived from untreated colon or colon treated with ethanol. Expression of α-actin, γ-actin calponin, caldesmon, smoothelin-A and tropomyosin mRNA in muscle strips from TNBS or DSS colitis was significantly increased compared to control animals. Similarly, expression of α-actin, calponin, caldesmon, smoothelin-A and tropomyosin protein as determined by western blot was significantly increased compared to control animals. We conclude that the expression of α-actin, γ -actin calponin, caldesmon, smoothelin-A and tropomyosin is upregulated in colonic circular smooth muscle from TNBS or DSS colitis. Increase in the expression of calponin, caldesmon and tropomyosin, which act to inhibit acto-myosin interaction, could contribute to decrease in smooth muscle contraction.

Transient intracellular elevations of Ca2+ are common signaling mechanisms used to allosterically regulate proteins. One potential target of Ca2+ is Ca2+/calmodulin dependent protein kinase type II (CaMK-II). CaMK-II is a multi-functional protein kinase known to influence cellular pathways such as cell motility and cell cycle progression. Within the cell cycle, CaMK-II promotes the expression of the regulator protein cyclin D1, which is necessary for cell cycle progression. To further understand CaMK-II’s role in cyclin D1 expression, the binding partners of cytosolic CaMK-II were studied using mass spectrometry. Several proteins were identified including β actin, β tubulins, tropomodulin- 3 and Flightless-I. Flightless-I was of the most interest because of its role as a transcriptional co-activator of β-catenin-dependent genes such as cyclin D1 and its ability to translocate out of the nucleus following serum stimulation or CaMK-II activation. For this study, I sought to determine whether the interaction between CaMK-II and Flightless-I mediates transcription of cyclin D1. First, Flightless-I was linked to green fluorescent protein (GFP) and live cell imaging was performed. Under serum stimulation or constitutive CaMK-II expression, GFP-Flightless-I was cytosolic. Serum starvation or inhibition of CaMK-II expression resulted in the localization of GFP-Flightless-I to the nucleus. Next, a luciferase based reporter gene assay was used to evaluate the effect of Flightless-I and CaMK-II on β-catenin-dependent transcription of cyclin D1. Over-expression of Flightless-I or inhibition of CaMK-II both resulted in decreased β-catenin-dependent transcription; whereas suppression of Flightless-I by siRNA enhanced transcription. Taken together, these results suggest a novel mechanism whereby the interaction between CaMK-II and Flightless-I influences gene transcription necessary for cell cycle progression.

Parkinson’s disease (PD) is a devastating neurodegenerative disorder characterized by progressive loss of dopaminergic neurons located in the substantia nigra. Accumulating evidence indicates that microglia-driven neuroinflammation contributes significantly to chronic neurodegeneration in PD. The matrix metalloproteinases (MMPs) play an important role in several neuroinflammatory paradigms; however, their relationship to dopaminergic neurodegeneration in PD remains relatively unexplored. To address this, the temporal relationship between MMP-2, MMP-3, MMP-9, and MMP-13 expression and dopaminergic neurodegeneration was compared in the 6-hydroxydopamine (6-OHDA) and lipopolysaccharide (LPS) models of PD. In dopaminergic SH-SY5Y cells, 6-OHDA treatment significantly increased MMP-13 mRNA expression; however, examination of 6-OHDA-lesioned rats demonstrated that MMP-13 protein expression does not change over the neurodegenerative time course, indicating that MMP-13 likely is not a major factor in neurotoxin-mediated neurodegeneration. MMP-3 and MMP-9 were not detected in either 6-OHDA model, but an increase in proMMP-2 expression was detected in nigral tissue samples 5 days post-surgery. This holds potential significance; however, active MMP-2 was not detected at any time point, suggesting that MMP-2-mediated proteolysis is not involved in 6-OHDA-induced neurodegeneration. In contrast to the 6-OHDA models, LPS triggered dramatic increases in MMP-2 and MMP-3 expression and activation that correlated with the neurodegenerative phase of this model. A significant increase in proMMP-9 was associated with this model, while no change in proMMP-13 was detected. In addition to MMP characterization, connective tissue growth factor (CTGF/CCN2), a protein that is transcriptionally and post-translationally regulated by MMPs was examined. In the LPS model, CTGF/CCN2 expression increased 5-fold and a protein fragment potentially representing cleaved CTGF/CCN2 was detected. The changes in CTGF/CCN2 occurred during peak increases in MMP-2 and MMP-3 activity. These experiments illustrate a strong temporal relationship between increased MMP expression and activation, elevated CTGF/CCN2 expression, and inflammation-induced dopaminergic neurodegeneration. There are clear differences between the 6-OHDA and LPS models that could hold significance for the pathogenesis of PD. Future studies aimed at interrupting MMP and CTGF function in the LPS model are required to further establish their role in inflammation-induced neurodegeneration and to assess their potential as a therapeutic target for the development of novel neuroprotective therapies for PD.

There are few published studies of larval fish assemblages from unregulated, tidal freshwater rivers. Patterns in the spatial and temporal distribution of larval fishes in the Mattaponi River were examined. Sampling took place on a weekly basis from February through August, 2006 and 2007. Larval fishes were categorized by taxa, reproductive guild, and residency guild. Group comparisons using multi-response permutation procedures (MRPP) indicated significant spatial and temporal differences in assemblage composition on multiple scales. Differences in assemblage composition were analyzed using non-metric multidimensional scaling (NMS). Interannual differences were attributable to anadromous and semi-migratory species. Seasonal differences were attributable to herrings, perches, and minnows. Both interannual and seasonal differences in assemblage composition may have been a result of changes in discharge. Spatial (i.e. longitudinal) variation of the larval fish assemblage differed by tidal regime. NMS and MRPP identified a distinct tidal freshwater larval fish assemblage. Tidal freshwater habitats may act as ecotones between marine and riverine ecosystems.

Hemoglobin-based oxygen carriers (HBOCs) offer a safe, more plentiful and long term alternate to blood banks. However, they have been found to increase blood pressure which can be attributed to an increase in total peripheral resistance (TPR). Lumenal nitric oxide (NO) scavenging by these HBOCs seems to be responsible for this hypertensive effect. In addition, it is believed that hemoglobin (Hb) tetramers and dimers may extravasate and consume additional nitric oxide in the perivascular and interstitial space. The purpose of the present study was to elucidate the role of NO scavenging and to confirm extravasation as a contributor to HBOC vasopressor effects in the spinotrapezius muscle. The present study investigated the vessel reactivity and mean arterial pressure response to three HBOCs: HBOC 201, HBOC 205 MW 400, and HBOC 205 MW 600. These varied in molecular weight (MW) and percentage of tetramers and dimers. It was found that larger polymers of HBOC showed no significant decrease in vasoactivity. Although larger polymers are less likely to extravasate, the remaining tetramers and dimers seem sufficient to contribute to the observed vasoactivity. Using NaNO2, a NO donor, in conjunction with the HBOCs almost completely abolished this hypertensive effect at higher concentrations. Further examination utilizing a nitric oxide synthase (NOS) inhibitor to mimic the HBOC vasopressor effects demonstrated that lower concentrations of NaNO2 were able to abolish the hypertensive effect. In vitro studies only further supported these results by demonstrating that NO consumption increases with HBOC dose. HBOC labeled with TRITC showed conclusive evidence that extravasation also plays a role in NO scavenging, even when minimal amounts of tetramers and dimers are present. In conclusion, the present study offers strong evidence that NO scavenging is responsible for the observed vasopressor effects. It also offers evidence supporting the theory that HBOC extravasation may be contributing to these vasopressor effects as well.

Schizophrenia is a debilitating disorder caused by the interaction of genetic and environmental factors. In this study, we identified candidate genes and single nucleotide polymorphisms from two genome-wide association studies, GAIN and CATIE. Nine SNPs representing four candidate genes were selected for replication studies with our Irish samples: Irish Case-Control Study of Schizophrenia (ICCSS), the Irish Study of High-Density Schizophrenia Families (ISHDSF), and the Irish Trio Study of Schizophrenia (ITRIO). In the ITRIO sample, rs4704591 (CMYA5 gene) showed nominal significance (p = 0.0447947). Combining ICCSS, ISHDSF, and ITRIO samples for rs4704591 increased sample size and power and yielded a p-value of 0.00388. This marker remained significant after Bonferroni correction for 9 markers genotyped in this study. CMYA5 gene binds to dysbindin protein in muscle. The dysbindin gene may influence glutamatergic neurotransmission, which has been suspected of being a mechanism by which the pathophysiology of schizophrenia is manifest. Our data suggest CMYA5 gene may be associated with schizophrenia in Caucasian subjects.