published_or_final_version / abstract / Biochemistry / Master / Master of Philosophy

Atherosclerosis is a chronic inflammatory disease thought to be initiated by endothelial cell dysfunction. Research described in this dissertation is focused on the role of zinc deficiency in endothelial cell activation with an emphasis on the function of the transcription factors nuclear factor-κB (NF-κB), peroxisome proliferator activated receptor (PPAR), and the aryl hydrocarbon receptor (AhR), which all play critical roles in the early pathology of atherosclerosis. Cultured porcine aortic vascular endothelial cells were deprived of zinc by the zinc chelator TPEN and/or treated with the NF-κB inhibitor CAPE or the PPARγ agonist rosiglitazone, followed by measurements of PPARα expression, cellular oxidative stress, NF-κB and PPAR DNA binding, COX-2 and E-selectin expression, and monocyte adhesion. Cellular labile zinc deficiency increased oxidative stress and NF-κB DNA binding activity, and induced COX-2 and Eselectin gene expression, as well as monocyte adhesion in endothelial cells. CAPE significantly reduced the zinc deficiency-induced COX-2 expression, suggesting regulation through NF-κB signaling. PPAR can inhibit NF-κB signaling. Zinc deficiency down-regulated PPARα expression and PPAR DNA binding activity in endothelial cells. Zinc deficiency compromised PPARγ transactivation activity in PPARγ and PPRE co-transfected rat aortic vascular smooth muscle cells. Furthermore, rosiglitazone was unable to inhibit the adhesion of monocytes to endothelial cells during zinc deficiency. Most of these effects of zinc deficiency could be reversed by zinc supplementation. An in vivo study utilizing the atherogenic LDL-R-/- mouse model generally supported the importance of PPAR dysregulation during zinc deficiency. LDLR-/- mice were maintained for four weeks on either zinc deficient or zinc adequate diets. Half of the mice within each zinc group were gavaged daily with rosiglitazone during the last stage of the study. Selected inflammation and lipid parameters were measured. The anti-inflammatory properties of rosiglitazone were compromised during zinc deficiency. Specifically, rosiglitazone induced inflammatory genes (MCP-1) in abdominal aorta only during zinc deficiency, and adequate zinc was required for rosiglitazone to down-regulate pro-inflammatory markers such as iNOS in abdominal aorta of the mice. Rosiglitazone significantly up-regulated liver IκBα protein expression only in zinc adequate mice. Plasma data also suggest an overall pro-inflammatory environment during zinc deficiency and support the concept that zinc is required for proper anti-inflammatory or protective functions of PPAR. Zinc deficiency also altered PPAR-regulated lipid metabolism in LDL-R-/- mice. Specifically, zinc deficiency increased plasma total cholesterol, and non- HDL (VLDL, IDL and LDL)-cholesterol. Plasma total fatty acids tended to be increased during zinc deficiency, and rosiglitazone treatment resulted in similar changes in fatty acid profile in zinc deficient mice. FAT/CD36 expression in abdominal aorta was upregulated by rosiglitazone only in zinc-deficient mice. In contrast, rosiglitazone treatment markedly increased LPL expression only in zinc-adequate mice. These data suggest that in this atherogenic mouse model treated with rosiglitazone, lipid metabolism can be compromised during zinc deficiency. AhR is another transcription factor involved in the development and homeostasis of the cardiovascular system. Cultured porcine aortic endothelial cells were exposed to the AhR ligands PCB77 or beta-naphthoflavone (β-NF) alone or in combination with the zinc chelator TPEN, followed by measurements of the AhR responsive cytochrome P450 enzymes CYP1A1 and 1B1. Zinc deficiency significantly reduced PCB77- induced CYP1A1 activity and mRNA expression, as well as PCB77 or β-NF-induced CYP1A1 protein expression, which could be restored by zinc supplementation. These data suggest that adequate zinc is required for the activation of the AhR-CYP1A1 pathway. Impairment of the AhR pathway presents an additional mechanism by which zinc deficiency negatively affects transcription factor function and homeostasis of the vascular system. Taken together, zinc nutrition can markedly modulate the pathogenesis of inflammatory diseases such as atherosclerosis.

Serine transhydroxymethylase (STHM), a pyridoxal 5'- phosphate requiring enzyme is indirectly involved in pyrimidine deoxynucleotide metabolism. A decrease in the activity of this enzyme could lead to altered deoxycytidine (dC) metabolism. This study was undertaken to determine if a vitamin B-6 deficiency affects dC metabolism. The effect of a vitamin B-6 deficiency on the activity of STHM in liver, thymus, spleen and bone marrow was examined. In addition, the effect of a vitamin B-6 deficiency on urinary excretion of dC was examined. The effect of a vitamin B-6 deficiency on the urinary excretion and tissue retention of ³H label from ip injected ³H-dC was monitored. Rats were assigned in groups of six to one of four treatment groups: ad libitum control (ALC), pair fed control (PFC), ad libitum deficient (ALD) or meal fed deficient (MFD). At the end of weeks 2 and 6, rats from each treatment group received an ip injection of ³H-dC. Urines were collected for 24 hours following the ip inhibited due to lack of cofactor, then dTMP levels would fall. In an attempt to increase the concentration of dTMP, enzymes active in the conversion of dC and dCMP to dUMP would be expected to increase. Thus, dC salvage pathways would increase and dC synthesis would decrease as metabolism shifts toward production of deoxythymidine triphosphate (dTTP). The result would be lower urinary dC excretion. The present study was undertaken to explore the relationship between vitamin B-6 and pyrimidine deoxynucleotide metabolism. There were four hypothesis tested: Vitamin B-6 deficient rats will excrete less urinary dC than either ad libitum or pair fed controls; vitamin B-6 deficient rats will excrete a lower percentage of labeled dC in urine than control rats; vitamin B-6 deficient rats will incorporate less labeled dC into DNA than control rats but may retain more label in tissues as dC metabolites; activity of STHM from tissues of vitamin B-6 deficient rats will be lower than that from the control rats. / Graduation date: 1990

Zinc status of Egyptian children 18-30 months and 6-10 years of age was characterized in relation to morbidity, growth, and socioeconomic variables. In a pilot study of children whose general nutrition ranged from adequately nourished to moderately malnourished, mean hair zinc was 135 ug/g (63-230 ug/g), with suboptimal zinc status suggested for 44%. Predictors of hair and serum zinc levels were explored for 23 school-aged and 40 preschool children. Included in models were weaning age for preschool children, body size (length- or height- and weight-for-age Z scores), current growth over 6 months or longer, illness experience over 10 to 12 months, demographic variables affecting food availability and distribution, sex, and season. Data were collected by Egyptian workers as part of a larger field project. Hair and serum samples were analyzed for zinc content by atomic absorption spectrophotometry. Results showed no difference in hair zinc levels by color, presence or absence of louse egg fragments and mucilage, or presence or absence of henna dye. In multiple regression models, the best predictor of hair zinc in preschool children was season of year, with zinc lower in summer. Season, negative effect of percent of weeks ill with diarrhea, and positive effects of socioeconomic status (SES) based on father's education/literacy and economic subsistence base excluding agriculture (ESB-A) predicted 23% of total hair zinc variation in preschool children. In preschool children serum zinc was lower in summer. Season, positive effect of rate of weight increase, and negative effects of rate of height increase, SES based on father's occupation(s) (SES2), and ESB-A predicted 53% of total serum zinc variation in preschool children. Serum zinc was higher in summer in school-aged children. Season, negative effect of SES2 and ESB-A, and positive effects of percent weeks ill with diarrhea and height for age Z scores predicted 60% of total serum zinc variation in school-age children. Negative effects of percent weeks ill with diarrhea and parents' age and child:adult ratio predicted 29% of hair zinc in school-aged children.

This study was conducted on a family of nine ranging in age from thirteen to fifty-two, to examine dark adaptation during physical activities.

Type-2 diabetes is a nationally growing health concern. Previous literature has implicated that vitamin D deficiency and type-2 diabetes are interconnected. At this time there are no guidelines in place to guide the evaluation or treatment of vitamin D deficiency in type-2 diabetic patients. In order to create up to date guidelines for the treatment and evaluation of vitamin D deficiency in type-2 diabetic patients an integrative literature review was conducted using EMBASE, PubMed, and CINAHL. The literature review resulted in 44 articles that met the inclusion and exclusion criteria. The literature review resulted in the creation of five clinical recommendations. The most significant change to current clinical standards includes screening all type-2 diabetic patients for vitamin D deficiency. Vitamin D supplementation is not recommended for use in treating type-2 diabetes, diabetic complications, or preventing the progression from prediabetes to diabetes. However, type-2 diabetic patients with vitamin D deficiency should be treated with vitamin D supplementation to improve indirect health outcomes and prevent morbidity and mortality.

SOCCER AS A MEANS OF REDUCING THE POPULATION MOVEMENT DEFICIENCY. Objectives: The first aim of this project is to study the impact of training on active amateur "A" team players of AFK Loko Chomutov. The second aim is to verify possibilities of using soccer in order to reduce population movement deficiency. Methods: In this project, methods of observation, measuring, comparing, and data analysis were used. Specifically, fitness prerequisites of individual players were assessed. The data analysis method was used in order to compare results of initial and final measuring within individual tests. Within the observed summer training period which lasted four weeks and contained sixteen training units (trainings of varied focuses with time span of 60 to 100 minutes), there have been assessed changes in fitness prerequisites in "A" team players of AFK Loko Chomutov. The research has been executed on 17 performance players between 22 and 33 years of age. Tested individuals have been engaged with football actively since their childhood. Outcomes: Thanks to application of specifically designed four-week training plan which included sixteen training units (trainings of 60 - 100 minutes) significant changes in fitness prerequisites in performance players have occurred. The training influenced speed, strength,...

Doctor of Philosophy / Division of Biology / Theodore Morgan / Temperature is a critical environmental parameter and thermal variation has significant effects on local adaptation and species distributions in nature. This is especially true for organisms that are isothermal with their environment. Variation in temperature imposes stress and directly influences physiology, behavior, and fitness. Thus, to thrive across a range of thermal environments populations must contain sufficient genetic variation, the capacity to respond plastically, or some combination of both genetic and plastic responses. In this work I first quantified patterns of phenotypic and genetic variation in nature and then dissected the genetic basis of variation in thermal traits. In the first aim I used natural populations of Drosophila melanogaster collected from a latitudinal transect in Argentina to investigate variation in heat stress resistance and cold plasticity within and among populations. I found heat stress resistance was highly variable within populations, but was strongly associated with the monthly maximum average temperature of each site. For cold plasticity I was able to demonstrate significant variation in plasticity within and among populations, however the among population variation was best explained by the altitude of each site. I hypothesized that this was caused by a difference in temperature fluctuations at high altitude sites relative to low altitude sites. To evaluate this hypothesis I paired our study with existing laboratory data that demonstrated significant fitness differences between high and low plasticity (and altitude) sites when these populations were reared in variable thermal environments. Thus, cold plasticity is an adaptive response to environmental variation. The final project focused on understanding the genetic basis of thermal variation. I fine-mapped a single co-localized heat and cold tolerance QTL via deficiency and mutant complementation mapping to identify four novel thermal candidate genes. There was no overlap of the deficiencies or genes associated with cold or heat stress resistance. Sequence analysis of each gene identified the polymorphisms that differentiate the lines. To test for independent associations between these polymorphisms and variation in nature the Drosophila Genome Reference Panel was used to confirm associations between allelic variation and cold tolerance in nature.

Master of Science / Food Science Institute - Food, Nutrition, Dietetics and Health / Weiqun Wang / Sorghum is a staple crop consumed in certain regions of Africa and Asia, where vitamin A deficiency is prevalent. However, the correlation of sorghum intake and vitamin A deficiency is inconsistent. The objective of this study was to identify and quantify the carotenoids and pro-vitamin A in the selected sorghum accessions with various pericarp pigments by using LC-MS. Among of total five carotenoids (α-carotene, β-carotene, lutein, zeaxanthin, and β-cryptoxanthin) that were identified and quantitated, three (α-carotene, β-carotene and β-cryptoxanthin) are precursors of vitamin A. The highest content of total carotenoids was detected in the sorghum accessions with yellow pericarp (PI656096, PI585374, PI563448, PI585351), while the highest β-carotene content was found in the accessions with brown or yellow pericarp (PI655996, PI656096, PI585374, PI563448, PI585351). The lowest carotenoids were found in the accessions with white pericarp (PI533943, PI656112, PI565121, PI560493). The pro-vitamin A was 584.9 ± 38.9 ng/g DW in yellow pericarp, 250.6 ± 28.9 ng/g DW in brown pericarp, and 89.0 ± 12.3 ng/g DW in white pericarp, respectively. It appeared the phenotypic diversity of sorghum pericarp colors was strongly associated with the contents of carotenoids and pro-vitamin A, indicating a different impact of various sorghum varieties on vitamin A deficiency and suggesting a possible prevention of vitamin A deficiency by breeding certain sorghum varieties with pericarp pigments.

The topic of the diploma thesis is micronutrients at home parenteral nutrition. In the theoretical part, parenteral nutrition is analyzed, including advantages and disadvantages, indications and contraindications. Intestinal failure is described as the most basic indication for the introduction of long-term / home parenteral nutrition. Following is the characterization and classification of micronutrients and a description of the consequences of their deficiency or surplus. Micronutrients are particularly important as cofactors and coenzymes in enzymatic reactions that would not be possible without them. The key question is whether the plasma levels show the actual state of the stock of the micronutrient in the organism, if the drop in most of the micronutrients that we find in patients reflects the actual deficiency to be substituted, or whether this decrease is an expression of their shift to other segments, the severity of the health condition. In the practical part of the diploma thesis are evaluated the levels of selected micronutrients (vitamin D, vitamin B12, folic acid, iron) in the group of patients with indication of home parenteral nutrition. The evaluation was based on laboratory analysis of patients' results. Key words: Parenteral nutrition, micronutrients, deficiency, surplus