Detection Capability of 89Sr and 90Sr Using Liquid Scintillation Counting.

ABSTRACT: We developed a new method for simultaneous determination of 89Sr and 90Sr with an emphasis on detectability. The samples were digested, and Sr was chemically purified followed by a single count on a liquid scintillation counter in three windows overlapping the 90Sr, 89Sr, and 90Y peaks. Gamma spectrometry was used to measure 85Sr, added for chemical recovery. The method was tested on 18 water samples spiked at levels from 9 to 242 Bq of 89Sr and 90Sr, with either single radionuclides or their mixtures. In addition, eight method blanks were measured. The data were analyzed numerically by solving a system of linear equations for 89Sr and 90Sr activities as analytes and 90Y activity as a participating component. The total uncertainties of the results were calculated numerically using variances and covariances. The average bias from the known activities was -0.3% (range from -3.6 to 3.1%) for 90Sr and - 1.5% (range from -10.1 to 5.1%) for 89Sr. The En-scores were within -1.0 and 1.0 at 95% confidence level. The detection capabilities of this method were determined by means of the decision threshold LC and the limit of detection referred to as the minimum detectable activity. All relevant uncertainties were propagated into the LC and minimum detectable activity. In addition, detection limits were calculated for the purpose of Safe Drinking Water Act monitoring. The detection capabilities were compared with the regulatory requirements in the US and EU for food and water. For samples spiked with either pure 89Sr or 90Sr, false positives were observed for the opposite radionuclide exceeding the above LC values. This was attributed to interference by the spiked activity. A new method was developed to calculate decision and detectability curves in the presence of interference.

Complement-Mediated Selective Tumor Cell Lysis Enabled by Bi-Functional RNA Aptamers.

Unlike microbes that infect the human body, cancer cells are descended from normal cells and are not easily recognizable as "foreign" by the immune system of the host. However, if the malignant cells can be specifically earmarked for attack by a synthetic "designator", the powerful effector mechanisms of the immune response can be conscripted to treat cancer. To implement this strategy, we have been developing aptamer-derived molecular adaptors to invoke synthetic immune responses against cancer cells. Here we describe multi-valent aptamers that simultaneously bind target molecules on the surface of cancer cells and an activated complement protein, which would tag the target molecules and their associated cells as "foreign" and trigger multiple effector mechanisms. Increased deposition of the complement proteins on the surface of cancer cells via aptamer binding to membrane targets could induce the formation of the membrane attack complex or cytotoxic degranulation by phagocytes and natural killer cells, thereby causing irreversible destruction of the targeted cells. Specifically, we designed and constructed a bi-functional aptamer linking EGFR and C3b/iC3b, and used it in a cell-based assay to cause lysis of MDA-MB-231 and BT-20 breast cancer cells, with either human or mouse serum as the source of complement factors.

Pseudomonas fluorescens strain CL145A - a biopesticide for the control of zebra and quagga mussels (Bivalvia: Dreissenidae).

Zebra mussels (Dreissena polymorpha) and quagga mussels (Dreissena rostriformis bugensis) are the "poster children" of high-impact aquatic invasive species. In an effort to develop an effective and environmentally acceptable method to control their fouling of raw-water conduits, we have investigated the potential use of bacteria and their natural metabolic products as selective biological control agents. An outcome of this effort was the discovery of Pseudomonas fluorescens strain CL145A - an environmental isolate that kills these dreissenid mussels by intoxication (i.e., not infection). In the present paper, we use molecular methods to reconfirm that CL145A is a strain of the species P. fluorescens, and provide a phylogenetic analysis of the strain in relation to other Pseudomonas spp. We also provide evidence that the natural product lethal to dreissenids is associated with the cell wall of P. fluorescens CL145A, is a heat-labile secondary metabolite, and has degradable toxicity within 24 h when applied to water. CL145A appears to be an unusual strain of P. fluorescens since it was the only one among the ten strains tested to cause high mussel mortality. Pipe trials conducted under once-through conditions indicated: (1) P. fluorescens CL145A cells were efficacious against both zebra and quagga mussels, with high mortalities achieved against both species, and (2) as long as the total quantity of bacterial cells applied during the entire treatment period was the same, similar mussel mortality could be achieved in treatments lasting 1.5-12.0 h, with longer treatment durations achieving lower mortalities. The efficacy data presented herein, in combination with prior demonstration of its low risk of non-target impact, indicate that P. fluorescens CL145A cells have significant promise as an effective and environmentally safe control agent against these invasive mussels.

Commandeering a biological pathway using aptamer-derived molecular adaptors.

Induction of molecular proximity can mediate a discrete functional response in biological systems. Therefore, creating new and specific connectivity between non-interacting proteins is a means of imposing rational control over biological processes. According to this principle, here we use composite RNA aptamers to generate molecular adaptors that link various 'target' molecules to a common 'utility' molecule, with the utility molecule being an entry point to a pathway conscripted to process the target molecule. In particular, we created a bi-functional aptamer that simultaneously binds to the green fluorescent protein (serving as a surrogate extracellular target) and the opsonin C3b/iC3b (serving as the utility molecule). This bi-functional aptamer enabled us to commandeer the C3-based opsonization-phagocytosis pathway to selectively transport an extracellular target into the lysosome for degradation. This novel strategy has the potential for powerful therapeutic applications with extracellular proteins involved in tumor development or surface markers on cancer cells as the target molecules.

gp38k (CHI3L1) is a novel adhesion and migration factor for vascular cells.

gp38k (CHI3L1) is a secreted heparin-binding glycoprotein whose expression, in vitro, is associated with vascular smooth muscle cell (VSMC) migration and invasion into the underlying gelatinous matrix. gp38k is expressed at high levels in postconfluent "nodular" VSMC cultures and at low levels in subconfluent proliferating cultures. In vivo, expression of gp38k homologs is high in regions of tissue remodeling and now has been detected in atherosclerotic plaques and in the developing heart. We tested the hypothesis that gp38k functions to modulate VSMC adhesion and migration. By use of modified Boyden chambers, gp38k at a concentration as low as 1 ng/ml has profound effects on VSMC migration but little or no effect on fibroblast migration. In addition, gp38k adsorbed to polystyrene surfaces directly promotes VSMC attachment and spreading. Attachment is inhibited in the presence of affinity-purified anti-gp38k or 10 mM EDTA. These results establish that gp38k is a new vascular cell adhesion and migration factor that may have a role in processes leading to vascular occlusion and heart development. gp38k may interact with VSMC via an EDTA-sensitive mechanism consistent with integrin mediated cell-matrix interaction.