The Clean Water Act, Water Quality, and Water Use

56 pages. Includes unsigned annotations by David Getches

Riverbank Filtration Impacts on Post Disinfection Water Quality in Small Systems—A Case Study from Auburn and Nebraska City, Nebraska

Small water systems can experience a fluctuating quality of water in the distribution system after disinfection. As chlorine is the most common disinfectant for small systems, the occurrence of disinfection byproducts (DBPs) represents a common problem for these systems. Riverbank filtration (RBF) can be a valuable solution for small communities located on riverbanks. The objectives of this study were to evaluate (i) the improvements in water quality at two selected RBF systems, and (ii) the potential lower concentrations of DBPs, in particular, trihalomethanes (THMs), in small systems that use RBF. Two small communities in Nebraska, Auburn and Nebraska City, using RBF were selected. Results from this study highlight the ability of RBF systems to consistently improve the quality of the source water and reduce the occurrence of THMs in the distribution water. However, the relative removal of THMs was directly impacted by the dissolved organic carbon (DOC) removal. Different THM concentrations and different DOC removals were observed at the two RBF sites due to the different travel distances between the river and the extractions wells

Direct affinity of dopamine to lipid membranes investigated by Nuclear Magnetic Resonance spectroscopy

Dopamine, a naturally occurring neurotransmitter, plays an important role in the brain’s reward system and acts on sensory receptors in the brain. Neurotransmitters are contained in lipid membraned vesicles and are released by exocytosis. All neurotransmitters interact with transport and receptor proteins in glial cells, on neuronal dendrites, and at the axonal button, and also must interact with membrane lipids. However, the extent of direct interaction between lipid membranes in the absence of receptors and transport proteins has not been extensively investigated. In this report, we use UV and NMR spectroscopy to determine the affinity and the orientation of dopamine interacting with lipid vesicles made of either phosphatidylcholine (PC) or phosphatidylserine (PS) lipids which are primary lipid components of synaptic vesicles. We quantify the interaction of dopamine's aromatic ring with lipid membranes using our newly developed method that involves reference spectra in hydrophobic environments. Our measurements show that dopamine interacts with lipid membranes primarily through the aromatic side opposite to the hydroxyl groups, with this aromatic side penetrating deeper into the hydrophobic region of the membrane. Since dopamine's activity involves its release into extracellular space, we have used our method to also investigate dopamine's release from lipid vesicles. We find that dopamine trapped inside PC and PS vesicles is released into the external solution despite its affinity to membranes. This result suggests that dopamine's interaction with lipid membranes is complex and involves both binding as well as permeation through lipid bilayers, a combination that could be an effective trigger for apoptosis of dopamine-generating cells

Hydrophobic Effects on Tyrosyl Ring 1H Chemical Shifts in Peptides

poster abstractHydrophobic environmental effects on tyrosine are measurable by 1H NMR spectroscopy and can allow us to detect interactions between peptides and lipid membranes. We first investigated the effects of hydrophobic environments on the 1H chemical shifts of tyrosine ring protons by using varying concentrations of isopropanol to mimic and calibrate the effects of hydrophobicity. Compared with this calibration, we then measured the interaction of tyrosine-containing peptides with sonicated unilamellar vesicles of phosholipids such as phosphatidylcholine and phosphatidylserine that are commonly found in biological membranes

Rates and Intermediates in Ser26 Mutants of Benzoylformate Decarboxylase

poster abstractBenzoylformate decarboxylase (BFDC), a thiamine diphosphate dependent enzyme, catalyzes decarboxylation of benzoylformate to benzaldehyde and CO2. The BFDC reaction proceeds through at least four individual chemical steps and, recently, NMR spectroscopy has been used to measure the ratios of intermediates in the overall reaction. This method permits calculation of rate constants for formation of the first intermediate, mandelylThDP (k2) and its subsequent decarboxylation (k3), as well as the combined breakdown of the enamine and product release (k4). As part of a study of the contributions of the active site residues, Ser26, His70 and His281, to the individual catalytic steps several Ser26 variants were expressed and purified. Initially, the variants were characterized using steady-state kinetics. Subsequently, the enzymes were mixed with benzoylformate and the mixture immediately acid quenched to trap intermediates of the reaction. NMR spectroscopy was used to identify and quantitate individual catalytic intermediates. Rate constants for the formation of these intermediates were then determined and compared to those of the wild-type enzyme. Here we report those results and discuss their implications for the role of Ser26 in the BFDC reaction mechanism

Lunar and Solar Torques on the Oceanic Tides

A general framework for calculating lunar and solar torques on the oceanic tides is developed in terms of harmonic constituents. Axial torques and their associated angular momentum and earth-rotation variations are deduced from recent satellite-altimeter and satellite-tracking tide solutions. Torques on the prograde components of the tide produce the familiar secular braking of the rotation rate. The estimated secular acceleration is approximately -1300 sec/century(sup 2) (less 4% after including atmospheric tides); the implied rate of change in the length of day is 2.28 milliseconds/century. Torques on the retrograde components of the tide produce periodic rotation variations at twice the tidal frequency. Interaction torques, e.g. solar torques on lunar tides, generate a large suite of rotation-rate variations at sums and differences of the original tidal frequencies. These are estimated for periods from 18.6 years to quarter-diurnal. At subdaily periods the angular momentum variations are 5 to 6 orders of magnitude smaller than the variations caused by ocean tidal currents

Fluorescence Measurements of Aromatic Amino Acids in the Presence of Lipid Membranes

Amphiphilic peptides are capable of finding their way to, and occasionally through, cellular membranes using a mechanism that includes specific amino acid sequences. Physical measurements of amino acid-lipid interactions are of interest for a quantitative description of peptide affinities to biological membranes. In this study, we investigate small peptide-lipid interactions using the fluorescence of the aromatic amino acids tyrosine (Tyr), tryptophan (Trp) and phenylalanine (Phe). Reference spectra in deuterated isopropanol solutions are obtained to mimic hydrophobic environments and are used to quantify the interaction of Lys-Tyr-Lys, Trp-Gly, and Gly-Phe with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and palmitoyl-oleoyl phosphatidylserine (POPS) lipid membranes. These fluorescence data complement previously reported UV absorption data and have the advantage of eliminating background and scatter from solution. Together with NMR data, these results can be used to more fully characterize lipid-aromatic amino residue interactions

DOMAIN STRUCTURE OF THE MAJOR ALLERGEN OVOMUCOID BY SOLUTION NMR

poster abstractThe interest in the ovomucoid protein is twofold. First it is a protein of interest for medical studies due to its potent allergen activity. Second, as a special variety of glycosylated protein (Kazal family), it allows us to explore the role of protein glycosylation in protein-membrane interactions for a particular, model case. The nature, location, and orientation of the glycosyl groups are determining factors in proteinmembrane interactions and therefore are critical to biological processes involving glycosylated proteins. We have found that as opposed to other glycosylated proteins, ovomucoid does not induce ionic currents across lipid membranes. This behavior likely has a structural cause, yet very little overall structural data is available. In this study, we use solution NMR spectroscopy to determine the structure of the chicken ovomucoid protein, taking advantage of the division of its structure into three stable domains of 55-65 amino acids each. We present results on the protein purification steps and isolation of separate domains suitable for solution NMR spectroscopy. We then present NMR results acquired on a 500 MHz spectrometer, and we show atomic models of individual domains and of overall protein structure from analysis of NMR spectra

Targeting the Role of Tyrosine in Amot Protein-Lipid Binding Events

poster abstractAngiomotins (Amots) are a family of adaptor proteins that have been shown to control cell proliferation and differentiation. Amots can selectively bind with high affinity to phosphoinositol containing membranes through the Amot coiled-coil homology (ACCH) domain. This binding event is linked to endocytosis, changes in cellular polarity, and apical membrane sequestration of nuclear transcription factors associated with development of cancerous phenotypes. Although the lipid selectivity of the protein has been well characterized, the residues involved in the ACCH domain binding these membranes have not been fully described. Understanding the structure-function relationship may provide pathways to modulate protein sorting and downstream signaling events inducing cellular differentiation, cancer cell proliferation, and migration. The fluorescent properties of the ACCH domain were previously used to characterize the binding event. However, the relative proximity of the five native tyrosines to the membrane may have led to differences in perceived lipid binding affinities based on fluorescence resonance energy transfer with fluorescently tagged lipids. A variety of short peptides correlating to the amino acid sequence of Amot surrounding these tyrosines were assayed and observed in different membrane mimicking environments. This was done to determine if each tyrosine had the ability to bury into the hydrophobic region of the membrane mimicked by the carbon chain lengths (alcohol study), or simply interacted with the hydrophilic head groups of the lipid (liposome study). In addition, the full length Amot80 ACCH domains (wild-type and tyrosine-to-phenylalanine mutants) were screened for trends in the varying environments. Interactions were characterized by shifts in maximum wavelengths for absorbance, excitation and emission peaks. A characterization of these shifts with respect to what is seen with the various tyrosine and phenalanine mutants may further our understanding of whether each tyrosine is buried within the protein or interacts with the head groups of the membrane