The goal of this dissertation is to investigate the interaction between cimetidine and dietary flavonoids using the Caco-2 cell transport model. It has been shown that flavonoids can change the bioavailability of pharmaceuticals, either by inhibiting metabolizing enzymes or inhibiting the drug efflux transporters. However, the effect of dietary flavonoids in the absorption of cimetidine has not been investigated. Therefore, the hypothesis of this study is that the absorption of cimetidine is mediated by a drug efflux pump, P-glycoprotein, of which dietary flavonoids can enhance the permeability of cimetidine by reducing P-glycoprotein function. The increase in permeability of cimetidine can increase the bioavailability of cimetidine. To test the hypothesis, three objectives were proposed. The first objective was to validate the Caco-2 transport model in our laboratory. The validation was performed by measuring the electrical resistance ofthe monolayer and determining the transport of paracellular marker. Also P-glycoprotein function was determined using rhodamine 123. The second objective was to describe the transport characteristics of cimetidine in the Caco-2 cell monolayers. The permeability of cimetidine was determined at different pH environments. When the permeability of cimetidine from apical to basolateral and basolateral to apical was compared, there appeared to be an effiux mechanism involved transport of cimetidine. The permeability of cimetidine in the presence of verapamil, a P-glycoprotein competitive inhibitor, suggested that P-glycoprotein was involved in the effiux. The third objective was to study the effect of dietary flavonoids on the permeability of cimetidine in the Caco-2 cell model. In the present study, four different flavonoids, quercetin, genistein, naringenin, and xanthohumol were selected. When co-treated with flavonoid aglycones, the permeability ofcimetidine was significantly reduced in the basolateral to apical direction. However, only genistin, a glycoside of genistein, significantly reduced the efflux of cimetidine. The present studies demonstrate that some dietary flavonoids, especially aglycones, can significantly reduce the effiux of cimetidine in the Caco-2 cell monolayers. Therefore, the fiavonoids consumed in a normal diet have the potential to enhance the bioavailability of cimetidine and possibly other P-glycoprotein substrates by altering their permeability. / Graduation date: 2004

Study Objective. To assess the dietary intakes of children with Attention Deficit Hyperactivity Disorder (ADHD); including macro- and micronutrient intake, the percentage of low nutrient density foods in the diet, as well as specific biochemical and anthropometric parameters. Design. Observational study. Setting. Provincial ADHD Program of BC, Children’s’ Hospital, Vancouver, BC. Sample. 44 children aged 6 - 12 years with ADHD, treatment-naïve or stable on medication for 6 months or longer. Main Results. ADHD Children were taller and heavier than population norms, and significantly taller than previously reported in ADHD. Of children aged 6-8 years, 28% were below the Estimate Average Requirement (EAR) for zinc and 61% of children aged 9-12 years were below the EAR for zinc. In addition, 28% of children aged 6-8 years and 39% of children aged 9-12 years were below the EAR for copper. Only 2% of children in the sample were below lab normal cutoffs for ferritin; however, 73% had serum zinc values below lab normal cut-offs and 23% had serum copper below lab normal cutoffs. Overall prevalence of serum zinc below the 2.5 percentile was 23% compared to 2% from National Health and Nutrition Examination Survey II (NHANES) data. Additionally, mean serum copper was significantly lower than NHANES II data. Mean energy intake was comparable to mean Estimated Energy Requirement (EER) based on the age, gender, height, weight and physical activity of subjects. In addition, mean dietary intake of Low Nutrient Density foods was not significantly different from NHANES II data and there was no significant difference in energy intake or the proportion of daily energy from protein, fat, and carbohydrate than what is observed from CCHS data. Medication treatment for ADHD was not associated with altered dietary intake or nutrient status. Conclusion. Results are suggestive of low zinc status in ADHD.

Three experiments were conducted to determine the efficacy of an <i>Escherichia coli</i> 6-phytase (Quantum phytase) in laying hens fed corn-soybean meal based diets. In experiment 1, the Escherichia coli 6-phytase (Quantum) was evaluated for its efficacy in a 40-wk laying hen production trial. A total of 1080 White Leghorn hens were fed mash corn-soybean meal (CSM) based diets containing 0.35%, 0.25% or 0.15% of non-phytate phosphorus (NPP) with the 0.25% and 0.15% diets containing 200, 400 and 600 U/kg of exogenous phytase. Only minor differences in production characteristics were found between the 0.35% and 0.25% treatments regardless of phytase addition, indicating that 0.25% NPP resulted in P intake that was at or above the hens requirement. In contrast, the hens fed the 0.15% NPP diet without phytase supplementation had significantly reduced production performance in comparison to the 0.35% treatment. The addition of phytase to the 0.15% diet improved these production characteristics to levels equal to or better than the 0.35% diet. The results indicated that Quantum phytase was efficacious in CSM-based diets fed to White Leghorn laying hens and can be used to reduce the need for diet supplementation with inorganic phosphorus. In experiment 2, the effect of Quantum phytase on nutrient digestibility and bone ash in laying hens fed CSM-based diets was investigated. A total of 108 White Leghorn hens were fed CSM-based diets containing 0.35%, 0.25% or 0.15% NPP with the 0.25% and 0.15% diets containing 200, 400 or 600 U/kg of exogenous phytase. A linear reduction in phytate digestibility, ileal protein digestibility and soluble P was reported with increasing levels of exogenous phytase in the 0.25% diet. Tibial bone ash percentage was higher in 61-wk-old hens fed 0.25% diets supplemented with 200 or 400 U/kg phytase. Overall, the Quantum phytase was not efficacious in improving nutrient digestibility in laying hens fed CSM-based diets deficient in NPP. In experiment 3, the impact of dietary Ca and P level on the efficacy of an E. coli-derived 6-phytase and the apparent digestibility of various nutrients was investigated in White Leghorn laying hens fed CSM-based diets. A total of 384 White Leghorn hens were fed CSM-based diets containing four levels of Ca (2.5, 3.5, 4.5 or 5.5%), two levels of NPP (0.15 or 0.30%), and two levels of phytase (300 or 600 U/kg feed). Increasing dietary Ca caused a decrease in AMEn, duodenal protein digestibility, Ca and phytate digestibility, percentage soluble P in feces and the percentage of poor quality eggs, while significantly increasing bone ash and hen-housed and hen-day egg production. The higher level of NPP (0.30%) decreased AMEn, fecal protein, Ca and P digestibility, and hen-housed and hen-day egg production, while increasing fecal soluble P and egg specific gravity in comparison to the lower NPP level. The higher level of dietary phytase (600 U/kg feed) significantly increased AMEn, phytate and P digestibility, soluble P in feces, and hen-day and hen-housed egg production, while significantly reducing the percentage of soft shelled, cracked and broken eggs. Overall, dietary phytase, Ca and NPP levels, either as main effects or in an interactive manner, can affect apparent nutrient digestibility and production traits in laying hens fed CSM-based diets.

Background: Few studies have evaluated habitual nutrient intake among HIV-infected youth in the United States, even though diet may influence disease progression and chronic complication risk. Objective: This study determined micro- and macronutrient intake in HIV-infected youth, and investigated relationships among nutrient intake, demographics, lipid profiles, and HIV-related factors. Design: HIV-infected subjects and healthy controls 1-25 years old were prospectively enrolled. Concomitant demographic, clinical and laboratory data were collected. Nutrient intake was assessed via 24-hr dietary recalls performed every 3 months for one year, analyzed with NDS-R software, and compared to Dietary Reference Intakes (DRIs) and Acceptable Macronutrient Distribution Ranges (AMDRs). Results: Subjects with ≥2 food recalls were analyzed (175 HIV+; 43 controls). Groups were similar in age, race, sex, body mass index, and kilocalorie intake. Both groups did not meet DRI for several micronutrients. HIV+ subjects had lower %DRI than controls for vitamins A, E, pantothenic acid, magnesium, calcium, folate and potassium. Percent kilocalories from fat was above normal and higher in HIV+ patients. Caloric intake was negatively correlated with current and nadir CD4 counts. Zinc, riboflavin, and magnesium %DRI were positively associated with current CD4+ count. In HIV+ subjects not on antiretroviral therapy, HIV-1 RNA levels were negatively correlated with protein intake. Conclusions: HIV+ youth have inadequate intake of several essential nutrients and poorer dietary intake compared to controls. Intake of some nutrients was associated with HIV-related factors. Further investigation is warranted to determine the impact of nutrition on HIV progression and chronic complication risk in this population.

The export of nitrogen (N) and phosphorus (P) from agricultural catchments is a major problem worldwide. The export of these nutrients is largely driven by storm events, and the hydrologic response of catchments varies within and between storm events. Antecedent soil moisture and rainfall rates have both been shown to affect the discharge and nutrient export from agricultural catchments, but their relationship to nutrient export is not fully understood. Currently, there are no studies that examine the leaching of both nitrate and phosphate from soil pools under the combined influence of differences in soil moisture and rainfall rates. The objectives of this study were to examine the combined effect of antecedent soil moisture and rainfall rates on the hydrologic response of soil and the export of nitrate and phosphate from the soil. The approach used intact soil monoliths in two experiments to first characterize the hydrologic response of the soil, and secondly to assess how the hydrologic response of the soil affects the leaching of nitrate and phosphate from soil pools. Differences in antecedent soil moisture and rainfall rates influenced both the amount of discharge and the hydrologic flow paths in the soil. As was expected, antecedent soil moisture governed the depth of discharge, with more discharge (runoff ratios= 0.89 to 0.91) produced by wet soil and the least runoff produced by dry soil (runoff ratios= 0.08 to 0.14) although this was not affected by the rainfall rate. Instead, rainfall rates predominantly affected hydrologic flow paths in the soil, with preferential flow at the beginning of the leaching period under high intensity rainfall (especially in wet soil), and predominantly matrix flow occurring under low intensity rainfall. The rainfall intensity did not appear to affect discharge volume. The mass of both nitrate and phosphate exported was higher under low intensity rainfall, ranging from 11.2 to 60.1mg/mU+00B2 and 77 to 4980μg/mU+00B2, respectively and from 0.9 to 34.4mg/mU+00B2 and 18.4 to 732μg/mU+00B2, respectively under high intensity rainfall. Antecedent soil moisture was significantly positively correlated with the depth of discharge produced, which also had a significant positive relationship with the mass of nitrate and phosphate exported (Spearman’s ρ= 0.75 to 0.81, p= <0.001), with greater masses of both nutrients exported from wet soil than dry soil. Soil moisture had contrasting influences on the nitrate concentrations in leachate, where nitrate concentrations and soil moisture were negatively related under low intensity rainfall and positively related under high intensity rainfall. Concentrations of phosphate in leachate were more variable, with no clear relationship to soil moisture, discharge, rainfall rate or soil phosphate pools. Antecedent soil moisture and the rainfall rate have a combined influence on the concentration of nitrate in leachate and an influence on the mass of both nitrate and phosphate exported. Although different hydrologic flow paths (matrix, preferential) were observed under the variable antecedent conditions and rainfall rates, this did not appear to affect nutrient fluxes from soil. This may be related to available nutrient pools and distributions in the soil in the current study. Understanding of the influence of flow types on the export of soil nutrient pools requires further study in a lab and a comparison of the breakthrough of nitrate and phosphate from soil pools with that of a conservative tracer (chloride). Nutrient and tracer breakthrough could then be compared to the hydraulic conductivity of the soil and the progression of the wetting front to fully understand the flow paths occurring and their effect on nutrient leaching.

Three experiments were conducted to determine the efficacy of an <i>Escherichia coli</i> 6-phytase (Quantum phytase) in laying hens fed corn-soybean meal based diets. In experiment 1, the Escherichia coli 6-phytase (Quantum) was evaluated for its efficacy in a 40-wk laying hen production trial. A total of 1080 White Leghorn hens were fed mash corn-soybean meal (CSM) based diets containing 0.35%, 0.25% or 0.15% of non-phytate phosphorus (NPP) with the 0.25% and 0.15% diets containing 200, 400 and 600 U/kg of exogenous phytase. Only minor differences in production characteristics were found between the 0.35% and 0.25% treatments regardless of phytase addition, indicating that 0.25% NPP resulted in P intake that was at or above the hens requirement. In contrast, the hens fed the 0.15% NPP diet without phytase supplementation had significantly reduced production performance in comparison to the 0.35% treatment. The addition of phytase to the 0.15% diet improved these production characteristics to levels equal to or better than the 0.35% diet. The results indicated that Quantum phytase was efficacious in CSM-based diets fed to White Leghorn laying hens and can be used to reduce the need for diet supplementation with inorganic phosphorus. In experiment 2, the effect of Quantum phytase on nutrient digestibility and bone ash in laying hens fed CSM-based diets was investigated. A total of 108 White Leghorn hens were fed CSM-based diets containing 0.35%, 0.25% or 0.15% NPP with the 0.25% and 0.15% diets containing 200, 400 or 600 U/kg of exogenous phytase. A linear reduction in phytate digestibility, ileal protein digestibility and soluble P was reported with increasing levels of exogenous phytase in the 0.25% diet. Tibial bone ash percentage was higher in 61-wk-old hens fed 0.25% diets supplemented with 200 or 400 U/kg phytase. Overall, the Quantum phytase was not efficacious in improving nutrient digestibility in laying hens fed CSM-based diets deficient in NPP. In experiment 3, the impact of dietary Ca and P level on the efficacy of an E. coli-derived 6-phytase and the apparent digestibility of various nutrients was investigated in White Leghorn laying hens fed CSM-based diets. A total of 384 White Leghorn hens were fed CSM-based diets containing four levels of Ca (2.5, 3.5, 4.5 or 5.5%), two levels of NPP (0.15 or 0.30%), and two levels of phytase (300 or 600 U/kg feed). Increasing dietary Ca caused a decrease in AMEn, duodenal protein digestibility, Ca and phytate digestibility, percentage soluble P in feces and the percentage of poor quality eggs, while significantly increasing bone ash and hen-housed and hen-day egg production. The higher level of NPP (0.30%) decreased AMEn, fecal protein, Ca and P digestibility, and hen-housed and hen-day egg production, while increasing fecal soluble P and egg specific gravity in comparison to the lower NPP level. The higher level of dietary phytase (600 U/kg feed) significantly increased AMEn, phytate and P digestibility, soluble P in feces, and hen-day and hen-housed egg production, while significantly reducing the percentage of soft shelled, cracked and broken eggs. Overall, dietary phytase, Ca and NPP levels, either as main effects or in an interactive manner, can affect apparent nutrient digestibility and production traits in laying hens fed CSM-based diets.

The objective of this study was to develop near infrared reflectance spectroscopy (NIRS) calibration equations from bamboo and fecal samples to predict diet composition and the physiological status of giant pandas. Discrimination between branch, culm, and leaf parts of bamboo resulted in an Rsquare (R2) of 0.88. The calibration equation for discriminating between 4 species of bamboo had an R2 of 0.47. Calibration equations were created for all bamboo species combined to determine the ability of NIRS to predict the nutrient constituents of CP, NDF, ADF, DM, and OM. No R2 was lower than 0.96, with the exception of DM at 0.63, which was consistently difficult to accurately predict due to variation in factors relating to difference in location of lab work (humidity, shipping, methods, etc.). Giant panda diets vary between seasons from eating primarily leaf to eating almost only culm. When bamboo part samples were compared between March and October, all resulting R2s were above 0.80. The sensitivity analyses for leaf and culm samples within diet season produced inconclusive results, but sensitivity analyses for fecal samples yielded an ability to more greatly discriminate between months that were further apart. For giant panda physiological status calibrations, fecal samples were collected from the Memphis Zoo, Smithsonian's National Zoo, Zoo Atlanta, and San Diego Zoo from 2006 to 2007. One-hundred fecal spectra were used to develop discriminant equations with which to predict between adults and juveniles. The resulting calibration was 100% correct for both age classes. Predictions between 252 male and female fecal spectra were 89% correct for females and 90% correct for males. A small number of samples (N= 60) were used to create a discriminant equation to differentiate between pregnant and non pregnant females. The exercise resulted in an R2 of 0.68 and a prediction of 100% for both pregnant and not-pregnant. It has been determined through these studies that NIRS has the potential to determine nutrient composition of bamboo and giant panda fecals, but increased sampling and equation development is needed before these calibrations are applicable in a captive or wild giant panda setting.

Bermuda has one of the highest concentrations of cave systems of any country in the world, but as the resident human population and tourism expand, this unique habitat is becoming increasingly threatened by development and water pollution. A water quality assessment was performed in six of Bermuda’s anchialine cave pools during summer 2007. Vertical water profiles were collected at each site to determine temperature, salinity, dissolved oxygen, and pH levels throughout the water column. Each cave pool had its own unique hydrological patterns. Additionally, water samples were collected at the surface and at the maximum depth to determine the concentration of nitrate, nitrite, ammonium, phosphate, and urea in the water. Further samples were collected to establish the fecal bacteria content in the surface waters. High nitrate levels were measured in two of the cave pools, while unsafe bacterial levels were measured in three pools. In situ nutrient addition experiments were used to examine effects of nutrient enrichment on algae naturally occurring in the six cave pool. Initial assessments of the pools determined distribution of foliose species. Six algal species (Ulva sp., Caulerpa mexicana, C. racemosa, C. sertularioides, Halymenia floresii, and Cryptonemia sp.) inhabited multiple sites, but not all species occurred at all sites. Changes in the biomass of individual plants were recorded for 6 to 12 days in response to N (nitrate), P (phosphate), and NP additions. The effect of nutrient addition on increase in biomass was significant over a 6 day period for Ulva sp. and H. floresii. Intermediate to no growth was seen in C. mexicana and Cryptonemia sp. C. mexicana exhibited both increases and in biomass depending on the site. Cryptonemia sp. had low but positive growth. A significant decrease in biomass occurred with C. racemosa and C. sertularioides. Primary productivity was measured using the traditional light dark bottle method. Differences in net productivity and respiration may explain why some plants responded positively and other negatively to nutrient additions. Ulva sp. and H. floresii showed potential as bioindicators for groundwater quality.

Benthic microalgae (BMA) living within the surface sediment of salt marshes are highly productive organisms that provide a significant proportion of organic carbon inputs into estuarine systems. BMA secrete extracellular carbohydrates in the form of low molecular weight carbohydrates and extracellular polymeric substances (EPS) as they migrate within the sediment. EPS plays an important role in the structure and function of BMA biofilms in shallow-water systems as EPS affects habitat structure, stabilizes the sediment, reduces sediment erosion, and is a carbon source for organisms. This study looked at the effect of nutrients and carbohydrate additions on BMA biomass, bacterial biomass, carbohydrate production, and glycosidase activity in the surface 5 mm of intertidal sediment in a subtropical salt marsh (Galveston Bay, Texas). Nitrogen and phosphorus were added to cores collected from the salt marsh and incubated in the lab over four days. Very little change was seen in the biomass of the benthic microalgae or in the different carbohydrate fractions with the added nutrients. The mean chlorophyll a concentration was 13 +/- 5 ug g-1 sediment, the mean saline extractable carbohydrate concentration was 237 +/- 113 ug g-1 sediment, and the mean EPS concentration was 48 +/- 25 ug g-1 sediment. The chlorophyll a and saline extractable carbohydrate concentrations initially decreased over the first 24 hours, but then increased over the rest of the experiment, indicating a possible species compositional shift in the BMA. With no major response with nutrient additions, it is likely that a different environmental factor is limiting for the growth of the benthic microalgae, and therefore the production of sEPS, in this salt marsh. A series of experiments was conducted in situ by adding glucose, alginic acid, and phosphorus to sediment within experimental plots. Samples were taken periodically over three to seven days to determine the biomass of the microbial community, enzyme activities and kinetics, and changes in the concentrations of several sediment carbohydrate pools. u-glucosidase activities (15 +/- 3 nmol g-1 h-1) were significantly higher than u-xylosidase (6 +/- 2 nmol g-1 h-1) and u-galactosidase (8 +/- 2 nmol g-1 h-1) activities within the sediment, and there was no suppression of u-glucosidase activity measured with the glucose addition. These data represent the first measurement of u- xylosidase and u-galactosidase activity in intertidal sediment dominated by BMA. Although preliminary experiments suggested a possible phosphorus limitation within the sediment, there was little change in the bacteria abundance or the benthic microalgae biomass when phosphorus was added in situ. This study begins to illustrate the dynamics of carbohydrate production and loss in this salt marsh, and the ability for the microbial community in the salt marshes of Galveston Bay to adjust to the nutrient and carbohydrate treatments.

Temporal variations of macroalgal cover and biomass (wet weight and dry weight) abundance and assemblage structure, and also the relationships between abiotic factors and assemblage structure, were studied in northeastern Taiwan (GPS: 25o06¡¦80¡¨N; 121o54¡¦60¡¨E¡^during 2001-2003. This study also investigated the causes affecting Gelidium natural resource and its management. Twenty-seven families with 67 species (5 families with 13 species for Chlorophyta, 3 families with 8 species for Phaeophyta, 19 families with 46 species for Rhodophyta) were recorded. The dominant species such as Pterocladiella capillacea and Polyopes polyideoides belong to temperate species. Macroalgal coverage, areal biomass, diversity, eveness and assemblage structure during 2001-2003 showed significant temporal changes with year and seasonal variations; however, the seasonal rhythm is changing between years. The seasonal pattern is different between 2001 and 2002/2003, biomass peaked in spring of 2002 and 2003. Pterocladiella capillacea, Polyopes polyideoides, Marginisporum crassissimum, Jania ungulate and Chondrus ocellatus contribute to temporal variations in assemblage structure. The results of multi-variate and uni-variate analysis suggest significant temporal variations in macroalgal assemblage structure in northeastern Taiwan, and the data from ANOSIM and SIMPER analysis demonstrated that the seasonal pattern was variable between years, in which it is coincident with seasonal variations in macroalgal abundance. Water temperature, summer high temperature, winter low temperature, nutrient and typhoon are attributable to temporal variations in macroalgal assemblage structure. Low temperature in winter, nutrient availability, and also typhoon restrict the natural resource of Gelidium in northeastern Taiwan; it is hypothesized that low P condition together with enhanced collection deep the decline of Gelidium resource, finally limiting the recovery of Gelidium population in next year. We found that the agar contents of Pterocladiella capillacea showed year and season variations with a negative relationship to biomass. Temporal fluctuations in agar contents was enhanced by low temperatures and high P but decreased by high N. Based on the observations, we conclude that over-collection in winter results in a decline of Gelidium resource in the coming years, thus it could be suggested that winter collection has to be controlled to maintain the sustainable utilization of Gelidium in northeastern Taiwan.