Delivery of secreted placental alkaline phosphatase (SEAP) gene in vitro and in vivo as a component of recombinant avian adenovirus (CELO)

Recombinant adenoviruses capable of expressing the gene of secreted placentary alkaline phosphatase (SEAP) under control of CMV-promoter was obtained on the basis of CELO avian adenovirus and human adenovirus-5 (Ad5) genomes. The efficiency of the CELO vector was determined in experiments with transduction of human (293, A549, and H1299), mouse (B16), and avian (LMH) cell cultures. It was shown in C57BL/6 mice in vivo that SEAP gene is expressed under conditions of intravenous, intranasal, and intratumoral application of recombinant adenovirus CELO-SEAP. The duration of expression of the alkaline phosphatase CELO = SEAP gene in immunocompetent mouse body was 21 days. The level of SEAP gene expression was measured in the allantois fluid of chicken embryo infected with recombinant adenovirus CELO-SEAP.     

Effect of oil-pollution of water on delayed fluorescence of the algae <Emphasis Type=Italic>Chlorella vulgaris</Emphasis> Beijer and survival rate of the cladoceran <Emphasis Type=Italic>Daphnia magna</Emphasis> Str.

The paper gives the results of the laboratory experiments on the influence of different concentrations of oil in water environment over the chlorella and daphnia. It is shown that as the oil content in water increases, the intensity of delayed fluorescence of test-culture of chlorella and the survival rate of the daphnia decrease. However, for the chlorella the decrease in fluorescence as the indicator of its photosynthetic activity is observed only at very high concentrations of oil (1000–2000 MPC), whereas the survival rate of the cladocerans begins to drop at 4 MPC. A similar dependence is observed when sunflower-seed oil is introduced into the environment as an imitator of the oil pollution. We suppose that the negative influence of oil over the microalgae and cladocerans is conditioned more by the physical factors than by chemical ones.     

Cultivation of reaggregated brain cells in rapidly rotating mini-rollers

An original method of cultivation of reaggregated brain cells with the aid of high-speed portable mini-rollers is described. The mini-roller consists of parallel rollers and an electric motor rotating the flasks at a speed of 60 to 70 rpm. The Moscona technique was used for preparing brain cell suspensions. During cultivation of the suspension of dissociated cells in high-speed mini-rollers, reaggregates with an internal organotypic structure were obtained. The method suggested provides stable and reproducible results.     

Multilayer associates based on oligonucleotides and gold nanoparticles

The development of nanodevices for the efficient transport of therapeutic molecules is one of the most urgent problems of modern molecular medicine. Noncovalent agents for the delivery of nucleic acids (NA) including those based on gold nanoparticles (GNPs) represent an attractive alternative to covalent systems, since it is easier in this case to provide the controlled release of NA. We have demonstrated the possibility to create potentially biocompatible associates of GNPs containing alternating layers of oligonucleotides and other polymers as a promising platform for the delivery of oligonucleotides into living cells. The multilayer (five layers) coated GNPs can be assembled by the sequential treatment of gold nanoparticles with nonthiolated oligonucleotide (ON), thiolated carboxyl-polyethylene glycol (SH-PEG3000-COOH), and linear polyethyleneimine (PEI). We have developed an algorithm for the analysis of multilayer coated GNPs by gel electrophoresis, photon correlation spectroscopy, and transmission electron microscopy. The assembly of associates bearing two oligonucleotide layers and having a net positive surface charge has been described. Multilayer coated GNPs were shown not to degrade in the presence of a high concentration of the major blood protein, serum albumin.