The need for a method to rapidly remove a radionuclide (UO₂²⁺) from environmental water samples has led to the development of hydrogel particles containing amidoxime groups for uranyl ion chelation. A miniemulsion polymerization technique using SDS micelles was employed for the preparation of the hydrogel as linked submicron particles. In polymerization, acrylonitrile was used as the initial monomer, ethylene glycol dimethacrylate as the crosslinker and 2-hydroxymethacrylate, 1-vinyl-2-pyrrolidone, acrylic acid, or methacrylic acid were added as co-monomers after the initial seed polymerization of acrylonitrle. The particles were characterized by transmission electron spectroscopy, scanning electron microscopy (SEM) and cryo-SEM. The amidoximated particles were superior to a commercially available resin in their ability to rapidly remove dissolved UO₂²⁺ from spiked groundwater samples.

The ability to modify/visualize biological surfaces, and then study the modified cell/virion in a range of in vitro and in vivo environments is essential to gaining further insight into the function of specific molecules or the entire entity. Studies of biological surface modification are generally limited to genetic engineering of the organism or the covalent attachment of chemical moieties to the cell surface(1,2). However these traditional techniques expose the cell to chemical reactants, or they require significant manipulation to achieve the desired outcome, making them cumbersome, and they may also inadvertently affect the viability/functionality of the modified cell. A simple method to harmlessly modify the surface of cells is required. Recently a new technology, KODE Technology has introduced a range of novel constructs consisting of three components: a functional head group (F), a spacer (S) and a lipid tail (L) and are known as Function-Spacer-Lipid or FSL constructs3. The spacer (S) is selected to provide a construct that is dispersible in water, yet will spontaneously and stably incorporate into a membrane. FSL construct functional moieties (F) so far include a range of saccharides including blood group-related determinants, sialic acids, hyaluronan polysaccharides, fluorophores, biotin, radiolabels, and a range of peptides(3-12). FSL constructs have been used in modifying embryos, spermatozoa, zebrafish, epithelial/endometrial cells, red blood cells, and virions to create quality controls systems and diagnostic panels, to modify cell adhesion/ interaction/ separation/ immobilization, and for in vitro and in vivo imaging of cells/virions(3-12). The process of modifying cells/virions is generic and extremely simple. The most common procedure is incubation of cells (in lipid free media) with a solution for FSL constructs for 1-2 hours at 37°C(4-10). During the incubation the FSL constructs spontaneously incorporate into the membrane, and the process is complete. Washing is optional. Cells modified by FSL constructs are known as kodecytes(6-9), while virions are kodevirions(10). FSL constructs as direct infusions and kodecytes/kodevirions have been used in experimental animal models(7,8,10). All kodecytes/kodevirions appear to retain their normal vitality and functionality while gaining the new function of the F moiety(7,8,10,11). The combination of dispersibility in biocompatible media, spontaneous incorporation into cell membranes, and apparent low toxicity, makes FSL constructs valuable research tools for the study of cells and virions.

Team-based care has consistently been associated with improved clinical outcomes. However, strategies for promoting and sustaining a team-based approach in family medicine practice are more elusive. We conducted a longitudinal time series cohort study of 30 primary care providers in seven practices to assess the sequential addition of three different chronic disease management feedback reports over 24 months, culminating in a team-based quality improvement intervention linked to feedback, assessing clinical performance and self-reported effectiveness. The proportion of patients at their low density lipoprotein target (<100 mg/dL) improved over the 24-month study period (P<0.001) but the rate of clinical improvement was more modest when feedback data were only presented at an individual and at a team level. When feedback reports were linked to a team-based quality improvement intervention, the results were more robust and were sustained for 12 months following the intervention cycle. Surveyed clinicians reported that the individual and team reports impacted both on their own practice approach and on team functioning. These findings suggest a strategic role for clinical performance feedback linked to team initiated quality improvement initiatives for improving both clinical outcomes and clinical team-based care.

Recombinant single-chain variable fragment antibodies (scFv) were specifically generated and selected for the measurement of environmental uranium with an antibody-based sensor. These sFvs, which recognized UO(2)(2+) complexed to 2,9-dicarboxyl-1,10-phenanthroline-acid (DCP), were produced using genetic material obtained from the spleen cells of rabbits immunized with UO(2)(2+)-DCP conjugated to keyhole limpet hemocyanin. Immunoglobulin light chain and heavy chain genes were amplified and cloned into the phagemid pSD3 for generation of a recombinant antibody library and phage-displayed antibodies. The screening process was designed to isolate antibodies that bound to a "loaded" noncovalent complex with high affinity, while selecting against binding to an "unloaded" complex. After five rounds of panning, individual positive scFv clones were used to infect E. coli TG1 and soluble scFv antibodies were purified and characterized. Binding studies showed that the best scFv bound tightly to the UO(2)(2+)-DCP complex (K(d), 19.6 nM). However, because of the depletion experiments performed on this library during the panning process, this scFv bound 1200-fold less tightly (K(d), 23.5 μM) to metal-free DCP. This scFv (clone 3A) was subsequently used to accurately determine the UO(2)(2+) concentrations in environmental water samples using a sensor based on kinetic exclusion analysis. The present studies demonstrate that recombinant scFvs with properties engineered for specific applications (i.e., biosensor-based measurement of metals in groundwater) can be prepared if the correct genetic material and techniques are employed. The phage display system permitted the generation of proteins with very specific binding properties (in this case, high affinity for a metal-chelate complex and low affinity for metal-free chelator). The recombinant scFvs isolated in these studies will be the basis for rapid and affordable assays for the detection of residual uranium in environmental water samples.

Field-based monitoring of environmental contaminants has long been a need for environmental scientists. Described herein are two kinetic exclusion-based immunosensors, a field portable sensor (FPS) and an inline senor, that were deployed at the Integrated Field Research Challenge Site of the U.S. Department of Energy in Rifle, CO. Both sensors utilized a monoclonal antibody that binds to a U(VI)-dicarboxyphenanthroline complex (DCPI in a kinetic exclusion immunoassay format These sensors were able to monitor changes of uranium in groundwater samples from approximately 1 microM to below the regulated drinking water limit of 126 nM (30 ppb). The FPS is a battery-operated sensor platform that can determine the uranium level in a single sample in 5-10 min, if the instrument has been previously calibrated with standards. The average minimum detection level (MDL) in this assay was 0.33 nM (79 ppt), and the MDL in the sample (based on a 1:200-1:400 dilution) was 66-132 nM (15.7-31.4 ppb). The inline sensor, while requiring a grounded power source, has the ability to autonomously analyze multiple samples in a single experiment The average MDL in this assay was 0.12 nM (29 ppt), and the MDL in the samples (based on 1:200 or 1:400 dilutions) was 24-48 nM (5.7-11.4 ppb). Both sensor platforms showed an acceptable level of agreement (9 = 0.94 and 0.76, for the inline and FPS, respectively) with conventional methods for uranium quantification.

OBJECTIVE: To create and test a slow-release antifibrotic drug-coated glaucoma drainage device using in vitro and in vivo experiments. METHODS: A slow-release device incorporating mitomycin C in poly(2-hydroxyethyl methacrylate) disks was developed using redox-polymerization techniques. A standardized preparation of this drug delivery device was attached to the Ahmed glaucoma valve (model FP7; New World Medical, Inc, Rancho Cucamonga, California). Semicircular disks (5 x 6 mm) of P(HEMA)-mitomycin C containing varying concentrations of mitomycin C per gram dry weight of the gel were attached to the lower half of an Ahmed glaucoma valve plate. Water was pumped through the modified Ahmed glaucoma valve at a rate comparable to that of aqueous humor outflow, and mitomycin C release was measured. Modified and unmodified Ahmed glaucoma valves were implanted in a rabbit model, and drug release and fibrosis were assessed after 3 months. RESULTS: The P(HEMA)-mitomycin C device released mitomycin C in vitro over 1 to 2 weeks. Studies in rabbits revealed that mitomycin C was released from the disks during the 3-month implantation. Histologic analysis demonstrated a significant reduction in inflammatory reaction and fibrosis in the resulting blebs. CONCLUSION: Our slow-release drug-coated glaucoma drainage device decreased fibrosis and inflammation in the resulting bleb in a rabbit model. CLINICAL RELEVANCE: This device could reduce the failure rate of glaucoma drainage devices.

Rotating-wall vessels are beneficial to tissue engineering in that the reconstituted tissue formed in these low-shear bioreactors undergoes extensive three-dimensional growth and differentiation. In the present study, bovine corneal endothelial (BCE) cells were grown in a high-aspect rotating-wall vessel (HARV) attached to collagen-coated Cytodex-3 beads as a representative monolayer culture to investigate factors during HARV cultivation which affect three-dimensional growth and protein expression. A collagen type I substratum in T-flask control cultures increased cell density of BCE cells at confluence by 40% and altered the expression of select proteins (43, 50 and 210 kDa). The low-shear environment in the HARV facilitated cell bridging between microcarrier beads to form aggregates containing upwards of 23 beads each, but it did not promote multilayer growth. A kinetic model of microcarrier aggregation was developed which indicates that the rate of aggregation between a single bead and an aggregate was nearly 10 times faster than between two aggregate and 60 times faster than between two single beads. These differences reflect changes in collision frequency and cell bridge formation. HARV cultivation altered the expression of cellular proteins (43 and 70 kDa) and matrix proteins (50, 73, 89 and 210 kDa) relative to controls perhaps due to hypoxia, fluid flow or distortion of cell shape. In addition to the insight that this work has provided into rotating-wall vessels, it could be useful in modeling aggregation in other cell systems, propagating human corneal endothelial cells for eye surgery and examining the response of endothelial cells to reduced shear.

This work demonstrated the feasibility of detecting divalent Cd(II) ions using antibody-modified microcantilever (MCL) sensors. Different surface modification methods were compared and multilayer approach was found superior than others for MCL surface modifications for antibody-based Cd(II) sensor development. When the Cd(II)-EDTA complex sample is injected into the fluid cell where the MCL is held, the MCL bends upon the recognition of the Cd(II) complex by the antibody on the surface of the MCL. Control experiments showed that complex that does not contain Cd(II) did not cause any bending of the MCL. The detection limit of the sensor was approximately 10(-9)M. The reaction rate analysis indicated that Langmuir adsorption model is appropriate to describe the absorption of Cd(II)-EDTA-BSA on the antibody-covered MCL surface.

The northeast Nile Delta region exhibits a high incidence of early-onset pancreatic cancer. It is well documented that this region has one of the highest levels of pollution in Egypt. Epidemiologic studies have suggested that cadmium, a prevalent pollutant in the northeast Nile Delta region, plays a role in the development of pancreatic cancer. Objective: We aimed to assess serum cadmium levels as markers of exposure in pancreatic cancer patients and noncancer comparison subjects from the same region in Egypt. Design and Participants: We assessed serum cadmium levels of 31 newly diagnosed pancreatic cancer patients and 52 hospital comparison subjects from Mansoura, Egypt. Evaluation/Measurements: Serum cadmium levels were measured using a novel immunoassay procedure. Results: We found a significant difference between the mean serum cadmium levels in patients versus comparison subjects (mean +/- SD, 11.1 +/- 7.7 ng/mL vs. 7.1 +/- 5.0 ng/mL, respectively; p = 0.012) but not in age, sex, residence, occupation, or smoking status. The odds ratio (OR) for pancreatic cancer risk was significant for serum cadmium level [OR = 1.12; 95% confidence interval (CI), 1.04-1.23; p = 0.0089] and farming (OR = 3.25; 95% CI, 1.03-11.64; p = 0.0475) but not for age, sex, residence, or smoking status. Conclusions: The results from this pilot study suggest that pancreatic cancer in the East Nile Delta region is significantly associated with high levels of serum cadmium and farming. Relevance to Clinical Practice/Public Health: Future studies should further investigate the etiologic relationship between cadmium exposure and pancreatic carcinogenesis in cadmium-exposed populations. Key words: cadmium, East Nile Delta region, environmental exposure, immunoassays, occupational exposure, pancreatic cancer, pollution.