AN ABSTRACT OF THE THESIS OF Nicole K. Jensen, for the Masters of Science degree in Forestry, presented on April 3, 2012, at Southern Illinois University Carbondale. TITLE: TREE-RING CHEMISTRY AND GROWTH RESPONSE TO EXPERIMENTAL WATERSHED ACIDIFICATION MAJOR PROFESSOR: Dr. Eric Holzmueller Forest ecosystems in the eastern United States are threatened by acid deposition rates that have increased dramatically since industrialization. We utilized two watersheds at the Fernow Experimental Forest in West Virginia to examine long-term effects of acidification on ecological processes. One watershed has been treated with ammonium sulfate (approximately twice the ambient deposition rate) since 1989 to simulate acid deposition while the other served as a control. Prior to treatment both watersheds were similar in age and species composition. Ten dominant overstory Prunus serotina and Liriodendron tulipifiera trees were selected and cored from each watershed to measure bolewood concentrations of elements essential for growth over time. In addition, changes in tree species basal area were analyzed utilizing 50 long-term growth plots established in 1990. Results of this experiment show lower calcium and magnesium concentration and increased acidic cation concentration in the treated watershed for both species indicating a negative treatment effect. These results were similar to that of a previous study conducted at the Fernow with periods of significant differences in cation concentrations between the treated and control watershed. Growth response, measured through relative growth rates of cored trees and changes in basal area from growth plots, was not as conclusive, but it did appear that treatment may be having a negative affect for both species during the last measurement of the growth plots. These results indicate a need for further research to understand the impact of long-term acidification so that sustainable forest management practices in areas affected by acid deposition may be developed.

The use of nitrogenous fertilizers in agricultural and forestry practices coupled with increased fossil fuel combustion and resulting nitrogen (N) deposition across the landscape have contributed to a near doubling of N inputs to terrestrial ecosystems.  With such dramatic changes have come adverse environmental consequences including the acidification of soil and water resources and an increased rate of biodiversity loss in both flora and fauna.  A method of rapidly predicting ecosystem susceptibility to N loss across large spatial scales would facilitate the identification of those systems most likely to contribute to potentially adverse environmental impacts.  To begin the development of such a framework, this research utilizes study sites located throughout the geographic ranges of Douglas-fir (Pseudotsuga menziesii) and loblolly pine (Pinus taeda) to explore relationships between hyperspectral remote sensing, N stable isotope ratios ("15N) and growth response to nitrogenous fertilizer.  In both species multiple linear regression models relating leaf-level reflectance to "15N showed strong predictive capabilities, with some models explaining more than 65% of the variance in "15N.  Significant correlations between "15N metrics and growth response to N fertilization were also observed in both species.  Additional exploratory analysis of the inclusion of "15N metrics with other environmental and edaphic variables to predict fertilizer growth response showed an increase in model performance with the addition of the enrichment factor (EF ="15NFol - "15NSoil).  This research demonstrates the ability of hyperspectral reflectance to predict "15N and reveals the potential of "15N to be included in future models to predict fertilizer growth response. / Master of Science

Introduction: Early growth response 1 (EGR1) is a zinc-finger transcription factor involved in the regulation of cell growth. It can act as either a tumour suppressor or a tumour promoter with a role in the induction of apoptosis in cancer cells by various pathways and is likely to play a role in colorectal cancer (CRC). EGR1 also appears to play a significant role in inflammatory pathways, therefore a possible role in Inflammatory Bowel Disease (IBD) is hypothesised. Patients with IBD have a greater risk of developing CRC, which is increased with duration of symptoms and severity of inflammation and dysplasia. The aim of this study is to determine whether EGR1 is differentially expressed in diseased colon tissue and to investigate novel EGR1-protein interactions in CRC cell lines. Methods: The relative EGR1 expression in CRC cell lines and in normal mucosa and tumours of colorectal cancer patients was determined by qRT-PCR. IBD patient samples were also examined for differential EGR1 expression levels by qRT-PCR, before and after stimulation with inflammatory mediators. Statistical analysis of the data was performed using ‘R’ statistical package, with the mixed-model ANOVA. Statistical significance was set at < 0.05. The genotype of three EGR1 variants was determined in the samples using PCR and sequencing, and the methylation status of regions of the EGR1 promoter was determined using bisulfite sequencing. A yeasttwo hybrid screen was conducted with EGR1 as bait, and screened against a SW480 CRC cell line library. Interesting novel interactions were investigated using immunocytochemistry and immunoprecipitation, as was the novel interaction between EGR1 and NOD2 and between EGR1 and components of the cytoskeleton. Results: Investigation into the relative EGR1 mRNA expression in CRC has shown that there is differential expression of EGR1 between matched normal mucosa and tumour. EGR1 expression is decreased in IBD patients compared with healthy controls. Induction of EGR1 by inflammatory stimuli also appears to be aberrant in these patients. The differential expression of EGR1 was not associated with aberrant methylation of a large region of the EGR1 promoter in either the CRC or IBD patients or with the genotype of EGR1 variants. EGR1 localises to both the cytoplasm and the nucleus in CRC cell lines and this study demonstrate interactions with the IBD susceptibility protein NOD2 and with components of the cyotskeleton. A yeast-two hybrid screen conducted with EGR1 as bait using a CRC cell line library has identified several other novel protein interactions of EGR1 in CRC cell lines. Conclusion: EGR1 is differentially expressed in both CRC and IBD, and in the case of IBD shows aberrant activity, suggesting that EGR1 may play a role in both colorectal diseases. EGR1 interacts with the IBD protein NOD2, and components of the cytoskeleton in CRC cells. Several novel protein interactions with EGR1 have been identified and warrant further study.

Secondary growth responses of native and nonnative trees exposed to the same climatic conditions can elucidate sensitivities and thus adaptability to a particular region. A long-term mixed-species planting in the pine barrens of northern lower Michigan presented an opportunity to discriminate responses from species commonly planted in this region. Mean ring-width chronologies from living native Pinus resinosa Ait. and P. strobus L. and nonnative P. sylvestris L. and Picea abies (L.) Karst. at this plantation were generated, standardized, and analyzed by correlation analysis against mean monthly climatic variables. The native pine chronologies had the highest mean ring widths and signal-to-noise ratios (SNR), were highly correlated to each other, and exhibited positive responses to years with above-normal April temperatures but no significant relationships to variations in precipitation. The P. sylvestris chronology was highly correlated to the other two pine chronologies and responded similarly to April temperatures but exhibited negative correlations to January and April precipitation and positive correlations to September precipitation. The P. abies chronology had the highest mean sensitivity and was correlated with the P. strobus chronology but only responded positively to precipitation from the previous December. The low SNR (P. sylvestris, P. abies), high mean sensitivity (P. abies), and larger number of significant correlations to variations in monthly climatic variables (P. sylvestris) suggest that these nonnative species are more sensitive to this local climate. These results provide insights to the adaptability, establishment, and geographic distribution of the nonnative Pinaceae.

The thesis deals with the effect of arbuscular mycorrhiza (AM) on the growth of selected plant species at a locality in České středohoří. This locality is characterized by close neighborhood of a semi-natural dry grassland and a former field abandonned in the 1990s, typical for the mosaic of biotopes in the region. The study is based on previous findings that some plant species, which are common at the semi-natural dry grasslands, do not colonize the abandoned fields. As AM is an important factor affecting diversity and productivity of plant communities we hypothesized that this phenomenon could be related to changes in AM fungal communities at the abandoned field. The hyphothesis was tested in a greenhouse pot experiment with three taxonomically related pairs of plant species, always one species growing at the abandoned field and the second one not. Growth and phosphorus uptake of the plants was followed in soils of both biotopes after factorial inoculation with AM fungal communities from both biotopes. The experiment was complemented by description of AM fungal communities in the roots of six plant species pairs from the locality using terminal restriction fragment length polymorphism (T-RFLP). The greenhouse experiment revealed positive mycorrhizal response in all plant species, but the origin...

For this dissertation, I examined the ability of field-grown plants to capture N presented in enriched patches or in whole-plant pulses. I assessed root proliferation in N-enriched patches when Agropyron desertorum plants had been previously fertilized or shaded. All plants responded with increased root growth rates in N-enriched patches. However, root proliferation by shaded plants was 50% less than unshaded plants. Unexpectedly, plants with higher N status had greater root growth rates in enriched patches than plants that had not received N supplement. I concluded that plants already under competitive pressure above ground for light and below ground for nutrients should be less able to respond to opportunities presented in nutrient patches. I then examined plant growth responses and biomass production of six Great Basin species (Bromus tectorum, Taeniatherum medusae, Agropyron desertorum, Pseudoroegneria spicata, Artemisia tridentata, and Chrysothamnus nauseosus) following a pulse ofN applied in the early, mid, or late spring. An equal quantity of N, applied continuously, was a control. Surprisingly, most of the species grown under the continuous supply had lower growth rates and less biomass production than plants recieving an N pulse. The exception was Chrysothamnus, which responded equivalently to all treatments. Generally, the greatest response occurred in early phenological stages. Four of the six species had their greatest response to the early-spring pulse, suggesting that these cold-season species are well-adapted to take advantage of early spring nutrient pulses. This study demonstrated that instead of benefitting from a season-long supply of N, there were times during the growing season when plants were able to use pulses of N for significant gains in biomass. I also investigated the root properties (root biomass, specific root length [the ratio of root length:root mass], and root uptake capacity) that determined plant response to pulses. Despite considerable temperature differences and changes in plant phenological stages, root uptake capacity remained remarkably constant throughout the season. However, this consistency did not explain the differences in productivity during the season. Root biomass also did not explain these growth responses to pulses. Instead, I suggest that the quantity of actively growing fine roots, plus the ability to effectively exploit the soil volume in the early spring, results in capture of early nutrient pulses.

Heart failure represents an important cause of death in Western Countries. The pathophysiology of heart failure is mainly associated with abnormalities in intracellular calcium control. We previously showed that Egr-1 negatively regulates expression of sodium-calcium exchanger (NCX) in vivo and in vitro. Here we tested the hypothesis that Egr-1 regulates expression of calcium storage proteins in the sarco-endoplasmic reticulum (SER), calsequestrin (CSQ) and/or ER, calreticulin (CRT) directly or indirectly via Egr-1:NFAT (nuclear factor of activated T-cells) formation. Secondarily, we hypothesized that this will reduce calcium mobilization. We found that undifferentiated 1293F cells, overexpressing Egr-1, have reduced CSQ compared to control H9c2 cells. We demonstrated that Egr-1 negatively regulates CSQ but not CRT expression. The Egr-1 mediated decrease in CSQ is linked to decreased calcium availability. Repression is by a novel NAB-independent (NGFI-A binding protein) activity localized to a.a. region 1-307. We conclude that Egr-1-mediated reductions in calcium storage protein expression alter calcium availability for cardiac contraction/relaxation.

Thesis (Ph. D.)--West Virginia University, 2007. / Title from document title page. Document formatted into pages; contains xv, 156 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 112-153).

All cells and organisms consume energy for survival. A robust system has evolved in vertebrates to serve as an energy reservoir. In particular, specialized cells, adipocytes, are responsible for the dynamic storage of energy by accumulating and releasing fatty acids. Fluctuating energy demands require adipose tissue to adjust in size, however complications can arise in both extremes giving rise to systemic diseases, such as obesity and diabetes mellitus (T2D). In mammals, leptin production in adipocytes is up-regulated by feeding and insulin to provide long-term post-prandial satiety. Although this regulatory connection is central to all physiological effects of leptin, the molecular mechanism remains unknown for leptin production. Here, we show that the transcription factor Egr1 is rapidly but transiently induced by insulin in adipose cells both in vitro and in vivo in a mTORC1-dependent fashion. Induction of Egr1 was immediately followed by an increase in leptin transcription. Chromatin immunoprecipitation and luciferase assays demonstrate that Egr1 directly binds to and activates the leptin promoter. Interestingly, the lipid droplet protein Fat specific protein 27 (FSP27) may work as a co-factor for Egr1 in regulating leptin expression. By using siRNA-mediated knock out of Egr1 along with its over-expression in adipocytes, we demonstrate that Egr1 is both necessary and sufficient for the stimulatory effect of insulin on leptin transcription. Knockout of the mTORC1-regulated translation repressor 4EBP1/2 increases leptin transcription both in vitro and in vivo. Adipose specific doxycycline-inducible constitutively active Rheb transgenic mouse lines contained higher circulating leptin and transcription of leptin following doxycycline treatment and were able to maintain elevated leptin levels following a 16 hour fast. Thus, insulin and nutrients, such as amino acids and glucose, activate leptin expression via the mTORC1-Egr1 regulatory axis.

There are a number of factors that influence crop yield, two of which are soil moisture and fertility.