Sequestration, fluorometric detection, and mass spectroscopy analysis of lanthanide ions using surface modified magnetic microspheres for microfluidic manipulation.

Several methods for rapid sequestration, fluorometric detection, and the subsequent mass spectroscopic analysis of lanthanide ions using surface modified polystyrene magnetic microspheres are demonstrated. Mixed-ligand antenna complexes of Eu(3+) in which one of the ligands is attached to the surface of the microspheres have been used as a means for the sequestration, immobilization, and detection of these ions. Using the ion-exchange properties of these microspheres, this scheme has been extended to the detection of nonluminescent ions. The principles of these assays form the basis for operation of a portable microfluidic device for general analytical and nuclear forensics applications and indicate the manner in which the established methods of analytical chemistry, such as liquid-liquid extraction and ion-exchange chromatography, can be adapted for such miniature devices.

A case study of cesium sorption onto concrete materials and evaluation of wash agents: Implications for wide area recovery.

To support the viability of a wash-down approach to mitigating nuclear contamination, this study presents a characterization of the aggregate of a common concrete by optical microscopy and the sorption-desorption characteristics of cesium from these into potential wash solutions. Various minerals with weathered surfaces displayed strong affinity for 137Cs with an effective partition coefficient Kd=120 mL/g for micas,>25-90 mL/g for feldspars, and>25-30 mL/g for amphiboles. The desorption Kd into 0.1M NH4Cl varied greatly but for amphiboles, sandstones, granite, and fine-grained quartzite it was>200 mL/g as a result of irreversible sorption. These same mineral phases are prevalent in all types of building materials, extending our conclusions more broadly to the problem of wide-area urban decontamination. In contrast, ionic solutions desorbed up to 98% of 137Cs from cement, suggesting that fresh concretes with an intact surface layer of cement could be more easily decontaminated if Cs+ interactions with the underlying minerals could be avoided. For practical applications common, non-hazardous chemicals such as sodium, potassium, and ammonium salts are as effective or more effective than harsher chemicals and expensive chelating agents. For example, when treated shortly after exposure, on time-scales commensurate with early response phase activities, 0.5M KCl could remove nearly 50% of bound 137Cs from concrete aggregate. Statistical analyses showed that desorption from the fine aggregate benefited from higher K+ and NH4 + concentrations. These results suggest that contamination in large areas of the urban environment can be dramatically reduced using common chemicals obtained readily from local stores.

Improvement of in vitro thrombolysis employing magnetically-guided microspheres.

Significant shortcomings in clinical thrombolysis efficiencies and arterial recanalization rates still exist to date necessitating the development of additional thrombolysis-enhancing technologies. For example, to improve tPA-induced systemic clot lysis several supplementary treatment methods have been proposed, among them ultrasound-enhanced tissue plasminogen activator (tPA) thrombolysis which has already found some clinical applicability. The rationale of this study was to investigate whether biodegradable, magnetic spheres can be a useful adjuvant to currently existing tPA-induced thrombolysis and further enhance clot lysis results. Based on an envisioned, novel thrombolysis technology--magnetically-guided, tPA-loaded nanocarriers with triggered release of the shielded drug at an intravascular target site--we evaluated the lysis efficiencies of magnetically-guided, non-medicated magnetic spheres in various combinations with tPA and ultrasound. When tPA was used in conjunction with magnetic spheres and a magnetic field, the lysis efficiency under static, no-flow conditions improved by 1.7 and 2.7 fold for red and white clots, respectively. In dynamic lysis studies, the addition of ultrasound and magnetically-guided spheres to lytic tPA dosages resulted in both maximum clot lysis efficiency and shortest reperfusion time corresponding to a 2-fold increase in lysis and 7-fold reduction in recanalization time, respectively. Serial microscopic evaluations on histochemical sections reconfirmed that tPA penetration into and fragmentation of the clot increased with escalating exposure time to tPA and magnetic spheres/field. These results delineate the effectiveness of magnetic spheres as an adjuvant to tPA therapy accelerating in vitro lysis efficiencies beyond values found for tPA with and without ultrasound. We demonstrated that the supplementary use of magnetically-guided, non-medicated magnetic spheres significantly enhances in vitro static and dynamic lysis of red and white blood clots.

Encapsulation and release of plasminogen activator from biodegradable magnetic microcarriers.

There are a number of therapies available to recanalize occluded arteries. However, even though proven beneficial, these approaches are not without significant shortcomings. Previous research showed that by encapsulating therapeutic thrombolytic enzymes in liposomic formulations, the reperfusion times in vivo were significantly lower than for administration of free thrombolytic. Like liposomes, biodegradable, diblock polymers of poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) have been shown to have therapeutic benefit as delivery vehicles for a variety of drug delivery concepts. We report on new formulations based on tissue plasminogen activator (tPA) encapsulated in magnetic, PLA-PEG microcarriers. We studied the tPA encapsulation efficiency, loading, and release after varying the molecular weight of polymer, carrier size, tPA solution composition, and use of ultrasound to enhance release. We loaded 3.3-9.4wt% tPA and 12-17wt% magnetite into the carriers, depending on the exact formulation. The release of tPA was complete 20min after reconstitution. Ultrasound insonation failed to enhance tPA release rates in smaller carriers but significantly enhanced release in larger carriers. With these formulations, we should be able to achieve lytic concentrations if we can magnetically concentrate 5mg of carrier within about 11ml of blood volume near the clot.

A novel human detoxification system based on nanoscale bioengineering and magnetic separation techniques.

We describe the conceptual approach, theoretical background and preliminary experimental data of a proposed platform technology for specific and rapid decorporation of blood-borne toxins from humans. The technology is designed for future emergent in-field or in-hospital detoxification of large numbers of biohazard-exposed victims; for example, after radiological attacks. The proposed systems is based on nanoscale technology employing biocompatible, superparamagnetic nanospheres, which are functionalized with target-specific antitoxin receptors, and freely circulate within the human blood stream after simple intravenous injection. Sequestration of the blood-borne toxins onto the nanosphere receptors generates circulating nanosphere-toxin complexes within a short time interval; mathematical modeling indicates prevailing of unbound nanosphere receptors over target toxin concentrations at most therapeutic injection dosages. After a toxin-specific time interval nanosphere-toxin complexes are generated within the blood stream and, after simple arterial or venous access, the blood is subsequently circulated via a small catheter through a portable high gradient magnetic separator device. In this device, the magnetic toxin complexes are retained by a high gradient magnetic field and the detoxified blood is then returned back to the blood circulation (extracorporeal circulation). Our preliminary in vitro experiments demonstrate >95% first pass capture efficiency of magnetic spheres within a prototype high gradient magnetic separation device. Further, based on the synthesis of novel hydrophobic magnetite nanophases with high magnetization ( approximately 55 emu/g), the first biodegradable magnetic nanospheres at a size range of approximately 280 nm and functionalized with PEG-maleimide surface groups for specific antibody attachment are described here. In future applications, we envision this technology to be suitable for emergent, in-field usage for acutely biohazard exposed victims as both the injectable toxin-binding magnetic spheres and the separator device are made to be portable, light-weight, zero-power, and self- or helper-employed. Details of the technology are presented and the state-of-knowledge and research is discussed.

Novel tandem column method for the rapid isolation of radiostrontium from human urine.

A method has been developed for the isolation of strontium from human urine for subsequent determination in sample volumes as low as 5-20 mL. This method involves the acidification of the sample using methanesulfonic acid and its decolorization using charcoal, treatment of the filtrate with Diphonix(®) resin, and subsequent concentration of strontium on Sr resin. Data from retention model simulations provided the initial conditions which were then optimized by actual column separations. Diphonix(®) resin was shown to be effective at removing alkali metal ions from the urine matrix under conditions that retain higher valence ions. The suggested processing method provides 99% recovery of Sr(2+), a concentration factor of 50, and an expected per sample processing time of less than 1 h.

The lysosomotropic agent, hydroxychloroquine, delivered in a biodegradable nanoparticle system, overcomes drug resistance of B-chronic lymphocytic leukemia cells in vitro.

Nonviral delivery systems are relatively easy to produce in the large scale, are safe, and elicit a negligible immune response. Nanoparticles (NPs) offer promise as nonviral vectors as biocompatible and -degradable carriers of drugs with targeting to specific sites by surface receptors of monoclonal antibodies (mAbs). We investigated the effect of four PEG-PLGA (polyethylene glycol-polylactic-co-glycolic acid) NP systems on drug-resistant B-chronic lymphocytic leukemia (B-CLL) cells in vitro, three of them encapsulating the drug, hydroxylchloroquine (HDQ), two with NP surface coatings of mAbs (NP1) CD20, (NP2) CD19, and CD20, and one (NP3) with no mAb, but tagged with the fluorescent marker, fluorescein isothiocyanate. The fourth NP system (NP4) was coated with anti-CD19/FITC and anti-CD20/Alexa-Fluor((R)) antibodies, but did not contain the active drug, HCQ. Our data indicate that PEG-PLGA nanoparticles with surface mAbs are suitable for selective drug delivery to B-CLL cells and produce a strong apoptotic effect when loaded with the lysosomotropic agent, HDQ.

Exploratory study on the effects of biodegradable nanoparticles with drugs on malignant B cells and on a human/mouse model of Burkitt lymphoma.

The aim of this study was to determine if Rituximab coated Biodegradable Nanoparticles (BNPs) loaded with Chlorambucil and Hydroxychloroquine could induce apoptosis of B-Chronic Lymphocytic Leukemia (B-CLL), MEC-1 and BJAB cells in vitro and evaluate their toxic and therapeutic effects on a Human/Mouse Model of Burkitt Lymphoma at an exploratory, proof of concept scale. We found that Rituximab-Chlorambucil-Hydroxychloroquine BNPs induce a decrease in cell viability of malignant B cells in a dose-dependent manner. The mediated cytotoxicity resulted from apoptosis, and was confirmed by monitoring the B-CLL cells after Annexin V/propidium iodide staining. Additional data revealed that these BNPs were non toxic for healthy animals, and had prolonged survival in this mice model of human lymphoma.

XAFS study of americium sorbed onto groundwater colloids.

The sorption of americium, as Am(III), onto groundwater colloids obtained from a marl aquifer was studied in 2 x 10(-2)M sodium bicarbonate groundwater and 2 x 10(-2)M sodium chloride bicarbonate-free solutions. At the in situ groundwater pH of 8.6, the americium was strongly sorbed onto the colloids. XAFS analyses were performed on these sorbed Am species to establish the oxidation state and its near-neighbour bonding. These XAFS data, obtained at 400 mg l(-1) colloid concentrations and total Am concentration of 1.53 x 10(-5)M (dissolved and onto colloids), indicated that Am remains trivalent, and that surface complexes are formed with the colloids without surface precipitation. This conclusion is based on the absence of Am-Am interactions in the second or third shells. The surface complexes generated by the Am(III) sorbed onto active sites are described on the basis of the XAFS data. They include the presence of about seven water molecules around the ternary surface complexes of this trivalent actinide.