Elemental Profile of Wheat in the Las Vegas market: Geographic Origin Discrimination and Probabilistic Health Risk Assessment.

This study investigates concentrations of toxic and potentially toxic elements (PTEs) in organic and conventional wheat flour and grains marketed in Las Vegas. Geographic origins of the samples were evaluated using Linear Discriminant Analysis (LDA). Monte Carlo Simulation technique was also employed to evaluate non-carcinogenic risk in four life stages. Concentrations of Al, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se, Sr, and Zn were determined using inductively coupled plasma mass spectrometry (ICP-MS) following hot block-assisted digestion. Obtained results showed non-significant differences in contents of toxic and PTEs between conventional and organic wheat grains/flour. Using LDA, metal(loid)s were found to be indicative of geographical origin. The LDA produced a total correct classification rate of 95.8% and 100% for US and West Pacific Region samples, respectively. The results of the present study indicate that the estimated non-carcinogenic risk associated with toxic element intakes across the four life stages were far lower than the threshold value (Target Hazard Quotient (THQ) >1). However, the probability of exceeding the threshold value for Mn is approximately 32% in children aged between 5 and 8 years. The findings of this study can aid in understanding dietary Mn exposure in children in Las Vegas.

Cyclodextrin-functionalized chromatographic materials tailored for reversible adsorption.

Novel dendronized silica substrates were synthesized. First- and second- generation polyaryl ether dendrons were appended to silica surfaces. Using Cu(I) mediated cycloaddition "click" chemistry, ß-cyclodextrin was tethered to the dendronized surfaces and to a nondendronized surface for comparison purposes. This synthesis strategy affords a modular, versatile method for surface functionalization in which the density of functional groups can be readily varied by changing the generation of dendron used. The surfaces, which are capable of adsorbing target analytes, have been characterized and studied using X-ray photoelectron spectroscopy (XPS) and vibrational sum frequency spectroscopy (VSFS). Fluorescence spectroscopy was used to study the surfaces' ability to retain coumarin 152 (C152). These studies indicated that the ß-cyclodextrin functionalized surfaces not only adsorbed C152 but also retained it through multiple aqueous washes. Furthermore, these observations were quantified and show that substrates functionalized with first-generation dendrons have a more than 6 times greater capacity to adsorb C152 than slides functionalized with monomeric ß-cyclodextrin. The first-generation dendrons also have 2 times greater the capacity than the larger generation dendrons. This result is explained by describing a dendron that has an increased number of ß-cyclodextrin monomers but, when covalently bound to silica, has a footprint too large to optimize the number of accessible monomers. Overall, both dendronized surfaces demonstrated an increased capacity to adsorb targeted analytes over the slides functionalized with monomeric ß-cyclodextrin. The studies reported provide a methodology for characterizing and evaluating the properties of novel, highly functional surfaces.

Clusters of ligands on dendrimer surfaces.

The development of methodology that is designed to allow a significant increase in the patterning and in the functionalization of the dendrimer is the ultimate goal of the research described here. Glycoside clusters based on TRIS were formed using click chemistry and were attached to PAMAM dendrimers. A series of dendrimers bearing tris-mannoside and an ethoxyethanol group was synthesized, and the binding interactions of these dendrimers with Concanavalin A were evaluated using inhibition ELISAs. The results of the inhibition ELISAs suggest that tris-mannoside clusters can replace individual sugars on the dendrimer without loss of function. Since tris-mannoside clustering allows for a redistribution of the dendrimers' surface functionalities, from this chemistry one can envision patterned dendrimers that incorporate multiple groups to increase the function and utility of the dendrimer.

Inhibition binding studies of glycodendrimer-lectin interactions using surface plasmon resonance.

Understanding protein-carbohydrate interactions is essential for elucidating biological pathways and cellular mechanisms but is often difficult due to the prevalence of multivalent interactions. Here, we evaluate the multivalent glycodendrimer framework as a means to describe the inhibition potency of multivalent mannose-functionalized dendrimers using surface plasmon resonance (SPR). Using highly robust, mannose-functionalized dithiol self-assembled monolayers on gold surfaces, we found that glycodendrimers were efficient inhibitors of protein-carbohydrate interactions. IC(50) values ranging from 260 nM to 13 nM were obtained for mannose-functionalized dendrimers with Concanavalin A.

Characterization of protein aggregation via intrinsic fluorescence lifetime.

Aggregation plays an integral role in multivalent protein-carbohydrate interactions, Alzheimer's and other amyloid-related diseases, and infection response. Efforts to apply controlled aggregation in toxin sensors have been made. We have developed a label-free intrinsic fluorescence lifetime assay that uniquely can monitor aggregation processes in real time without interference from precipitation. Fluorescence decay curves were measured with high precision at 1 s time intervals following addition of a glycodendrimer to a lectin-containing solution. Changes in the fluorescence intensity and lifetime signified formation of complexes. However, these changes were not associated with the initial lectin-sugar binding events. Rather, they appeared to be caused by clustering and subsequent conformational rearrangement of the lectins. Studies were conducted with mannose-functionalized polyamidoamine (PAMAM) dendrimers of the second through sixth generations and Concanavalin A. The apparent rate constant, when expressed on a per-mannose basis, increased with dendrimer generation, particularly in going from the fourth to the sixth generation. However, the identical fluorescence decay waveforms for saturating amounts of dendrimer suggested that all of the glycodendrimer generations studied reach a comparable state of aggregation. Although self-quenching of tryptophan resonances that was induced by clustering was monitored in this study, the reported method is not limited to such and is viable for numerous binding studies.

Binding of mannose-functionalized dendrimers with pea (Pisum sativum) lectin.

Lectins are invaluable tools for chemical biology because they recognize carbohydrate arrays. Multivalent carbohydrate binding by lectins is important for processes such as bacterial and viral adhesion and cancer metastasis. A better understanding of mammalian lectin binding to carbohydrate arrays is critical for controlling these and other cellular recognition processes. Plant lectins are excellent model systems for the study of multivalent protein-carbohydrate interactions because of their robustness and ready availability. Here, we describe binding studies of mannose-functionalized poly(amidoamine) (PAMAM) dendrimers to a mitogenic lectin from Pisum sativum (pea lectin). Hemagglutination and precipitation assays were performed, and results were compared to those obtained from concanavalin A (Con A), a lectin that has been studied in more detail. Isothermal titration calorimetry (ITC) experiments are also described.