In this study we describe a novel sensor system to detect toxic chemicals based on measurement of the quantity of Saccharomyces cerevisiae P450 mRNAs induced by them. Detection was conducted using a flow-injection-type sensor system based on surface plasmon resonance (SPR). The DNA and peptide nucleic acid (PNA) probes containing a complementary sequence to a part of P450 mRNA were immobilized on the sensor chip and the P450 mRNAs hybridized to the probes were quantified. We succeeded in detecting 10 ng/L (10 ppt) of atrazine using both DNA and PNA probes. Using this sensor system, we were able to detect bisphenol A in addition to atrazine. Furthermore, we achieved higher sensitivity by amplifying the target P450 mRNA based on nucleic acid sequence-based amplification (NASBA). This method allows for sensitive, rapid, and easy detection of some toxic chemicals.

Atrazine↗

PubMedDate not supplied

In vivo expression of human growth hormone by genetically modified murine bone marrow stromal cells and its effect on the cells in vitro.

Human growth hormone (hGH) is frequently used clinically for growth abnormalities in children and also in adults with growth hormone deficiency. The hormone is usually administered to the individuals by frequent injections. In the present study we investigated the potential of bone marrow stromal cells as vehicles to deliver the GH in vivo by infusion of cells transduced with hGH cDNA into mice femurs. The effect of the hormone on the transduced cells in vitro was also assessed. Bone marrow stromal cells established from a mouse model of human osteogenesis imperfecta mice (oim) were transduced with a retrovirus containing hGH and neomycin resistance genes. The hGH-expressing cells were selected in a medium containing G418 and were then assessed for the hGH expression in vitro. The selected cells synthesized 15 ng/10(6) cells of hGH per 24 h in vitro and exhibited alkaline phosphatase activity when they were treated with the human recombinant bone morphogenetic protein 2 (rhBMP-2). The transduced cells also proliferated faster than the LacZ transduced cells but they did not exhibit a higher rate of matrix synthesis. When 2 x 10(6) hGH+ cells were injected into the femurs of mice, hGH was detected in the serum of the recipient mice up to 10 days after injection. The highest level of growth hormone expression, 750 pg/ml, was detected in the serum of the recipient mice I day after injection of the transduced cells. hGH was also detected in the medium conditioned by cells that were flushed from the femurs of the recipient mice at 1, 3, and 6 days after cell injection. These data indicate that bone marrow stromal cells could potentially be used therapeutically for the delivery of GH or any other therapeutic proteins targeted for bone. The data also suggest that GH may exert its effects on bone marrow stromal cells by increasing their rate of proliferation.

Alkaline Phosphatase↗