[The interaction between nerve cells and carbon nanotube networks made by CVD process investigation].

In this research we investigate neuroblastoma cells cultivated on single-walled carbon nanotubes networks made by CVD method on silicon substrates. The complex analysis of grown cells made by atomic force, electron microscopy and Raman spectroscopy was carried out and the effect of nanotube growth process on proliferation factor was investigated. It is shown that despite of a weak decrease in proliferation, cell morphology remains unchanged and no physical or chemical interaction between carbon nanotubes and cells is observed. The results of the research can be used to investigate the interaction between conductive nanomaterials and cells for the development of neural replacement implants. Also they can be useful in bio-electronic interface investigation of signal propagation in neurons.

[Cultivation of continuous cell lines on the substrate of carbon nanotubes and the effect of electric stimulation on cell proliferation].

It was demonstrated that the three studied samples of carbon nanotubes of domestic production fixed on the substrate surface did not have toxic effect and could be used for cell cultivation. A biocompatible conductive coating based on carbon nanotubes and bovine serum albumin was developed. The efficacy of the coating for growing in vitro cell cultures was tested. A device was developed for electric stimulation of the cells. Local electric potential was applied to the cells using nanoscale electrodes. The results of human embryonic fibroblast cultivation in a pulsed electric field on conductive nanocomposite substrates were presented. An 26% increase in the proliferative activity of cells was observed at potentials up to 100 mV.

[The morphological and karyological characteristics of MDCK and vero (B) cells cultures on plant hydrolyzate-based nutrient media].

MDCK and Vero (B) cell cultures were propagated during 10 passages in the experimental nutrient media containing the soybean powder hydrolyzate prepared using trypsin and bromelain enzymes and the rice powder hydrolysate prepared with trypsin and in the control DMEM and SFM4 MegaVir media. The karyological, morphological, and proliferative characteristics of continuous cultures were examined and compared. The experimental media supplied with 3% fetal bovine serum (FBS) (Gibco, U.S.A.) showed high growth-enhancing properties and failed to affect their morphology. After propagated during 10 passages in the experimental media preserved a stable karyotype. MDCK cell cultures in the nutrient media based on rice and soybean powder hydrolyzates low (2%) in FBS caused no substantial changes in the proliferation indices and morphological and karyological characteristics of cell cultures.

[Influenza virus reproduction in the MDCK cells adapted to growth in serum-free Hybris-2 medium].

Whether the MDCK cell line might adapt to grow in serum-free Hybris-2 medium and influenza viruses might be reproduced in the adapted cells was studied. Seventeen passages using the Hybris-2 medium yielded cells adapted to growth in this medium (MDCK-BS). The reproduction of influenza A (H1N1 and H3N2) and B viruses versus the cells cultured in Eagle's medium was studied. The laboratory strain of influenza A/Aichi/1/68 (H3N2) and the strain B/Ohio/01/05 of influenza B in equal titers were shown to be reproduced in both control cells on Eagle's medium and MDCK-BS cells adapted to growth in the Hybris-2 medium. The reproduction of the strains A/Brisbane/10/07 (H3N2) and A/Solomon Islands/3/06 (H1N1) was less active in the MDCK cells. Each strain of influenza viruses displayed varying infective activities. The developed serum-free Hybris-2 medium may be used for cultivation of monolayer continuous MDCK cells and for their reproduction of influenza A and B viruses.

[Vitaherpavac is the first Russian herpes simplex virus vaccine obtained on the Vero B continuous cell line].

Vitaherpavac, a dry inactivated herpes simplex virus (HSV) culture vaccine, has been obtained, by using the Vero B continuous cell line as a substrate for accumulation of herpes simplex virus types 1 (US strain) and 2 (VN strain). Vitaherpavac and the similar vaccine Herpovax made by the Research Institute of Vaccines and Sera, Saint Petersburg (for which preparation a primary trypsinized chick embryo cell culture used as a substrate for accumulation of HSV types 1 and 2), underwent comparative clinical trials. The tolerability and therapeutic effectiveness of the vaccine were tested in patients diagnosed as having chronic frequently recurring herpes. The trials have yielded positive results that suggest that it is expedient to introduce of the new vaccine Vitaherpavac into practice to treat chronic recurrent herpetic infection of various localizations. Vitaherpavac has been registered in the Russian Federation and permitted for medical application.

Effect of exogenous frequency exposure on human cells.

Effects of exogenous bioresonance oscillations on biological and morphological characteristics of continuous human lung and liver cells were studied. Proliferative activities and viability of cells decreased after exposure to frequencies of 8 and 78.5 Hz. This was paralleled by cytopathic disorders in lung and liver cells. The frequency of 72 Hz exhibited a less intense effect on both cell types. Cell contamination with mycoplasmas was provisionally suppressed after cell exposure according to F44 and F45 programs and to 97.5 and 69.5 Hz frequencies.

[In vitro and in vivo studies of the "VITA" device]. / Izuchenie pribora "VITA" v éksperimentakh in vitro and in vivo.

Many in vitro and in vivo experiments using contemporary methods supported safety and efficiency of "VITA" device protecting humans and living beings from electromagnetic fields. Therefore, bioenergetic safety device "VITA" is advisable for use.

[The use of Vero (B) cell line for medico-biological preparations]. / Liniia kletok Vero (B) dlia prigotovleniia mediko-biologicheskikh preparatov.

A variant of Vero cell line has been obtained, adapted to the Russian Eagle's medium with 8% fetal calf serum. The variant is characterized by intensive cell proliferation, possesses a different content of chromosomes, contains no oncogens and contaminants, specifically, Mycoplasma. The strain was characterized and certified in accordance with the WHO requirements. Inoculation (110 ampules) and working (100 ampules) stocks at the levels of the 170th and 178th passages were placed for storage in liquid nitrogen. Vero (B) cell line is sensitive to herpes simplex virus types 1 and 2, CMV, hepatitis A virus, recombinant variolovaccinia strain expressing HbS antigen, etc. The line retains its biological properties from passage 178 to passage 200 and is recommended for control and preparation of various biomedical agents, including antiviral vaccines.

[Use of the DNA-specific fluorochrome olivomycin for work with cell cultures]. / Ispol'zovanie DNK-spetsifichnogo fliuorokhroma olivomitsina dlia raboty s kletochnymi kul'turami.

A high-sensitive, easy and rapid proximate method has been developed to reveal mycoplasmas in the monolayer cell cultures using home fluorescent antibiotic olivomycin. This method has been used to screen many cell lines with its high effectiveness being shown. As based on this method the following control methods are worked out: indication of mycoplasmas in animal blood sera, used for the cultivation of cell lines detection of contaminant microorganisms of the cell cultures (fungi and bacteria) and human ureaplasmas.

[Cytomorphologic study of a line of hybrid somatic cells persistently infected with the scrapie agent]. / Tsitomorfologicheskoe issledovanie linii gibridnykh somaticheskikh kletok, persistentno infitsirovannykh agentom skrepi.

Virological, cytological and electron microscopic methods were used to study the peculiarities of the scrapie agent persistence in the tissue culture of the mouse and human hybrid cells. A long-term persistence of the scrapie agents in the cells (658 days) has been obtained. The fact of persistence is confirmed by the results of biotest and electron microscopic studies of the mouse CNS. The agent persistence promotes a decrease in the mitotic activity of the infected cells and development of the ultramicroscopic changes in cells similar to the picture of the specific changes in the CNS of mice inoculated by scrapie agent.

[Morphological characteristics of the brain lesions in mice infected with a homogenate of L cells latently infected with the scrapie agent]. / Morfologicheskie osobennosti porazhenii mozga myshei, zarazhennykh gomogenatom kletok L, latentno infitsirovannykh agentom skrepi.

Degeneration of neurons of the Ammon horn lower branch both in the early and terminal stages of the disease of mice infected with the homogenate of L cells latently infected with the scrapie agent (the L-S system) was frequently detected alongside with brain lesions typical of slow infections (vacuolation). Examinations of chromosomes in metaphase plates of L-S cells carried out by several methods including the TAC system for texture analysis of the image (Leutz, BRD) revealed three marker chromosomes new for continuous L cells, the appearance of true chromatid translocations as well as significant changes in chromosome numbers. Besides, ultrastructural features of L-S cells at later stages of cultivation were revealed. It is assumed that the active effect of the scrapie agent on L cells infected with it resulted in the emergency of a new antigen capable to induce selective affection of the neurons of the Ammon horn lower branch in susceptible mice.

In vitro studies with the scrapie agent.

Persistent infection with the scrapie agent has been established in L cells. The agent was propagated in homogenates of the cell line designated L-S. The L-S cells showed slower growth rate and morphological changes like pycnosis of nuclei and vacuolization of their cytoplasm. Cytogenetic analysis revealed rearrangement of chromosomes in L-S cells. In the course of passaging the number of cells with the characteristic marker chromosome decreased. Along with this cells were found with deletion of one arm of the marker chromosome. In addition, 3 new marker chromosomes were detected in infected cells, suggesting the influence of the scrapie agent on cytogenetic processes in scrapie-carrier cultures. The infectious activity of nucleic acids isolated from L-S cells was determined in BALB/c mice inoculated with untreated, DNase-treated and pronase-treated nucleic acid preparations. A slightly decreased infectious activity has been noted after DNase and pronase treatments.

[Karyological characteristics of continuous mouse cell lines infected with the scrapie agent]. / Kariologicheskaia kharakteristika perevivaemykh linii myshinykh kletok, infitsirovannykh agentom skrepi.

A karyological study of murine cell line L, latently infected with different strains of the scrapie agent (Compton and C-506) showed that the both variants number of chromosomes and the number of Robertson's translocations of chromosomes decrease insignificantly with an increase in the time of subcultivation. The use of the C-method of differential chromosome staining revealed four new analogous chromosomes in both experimental cell lines. This phenomenon is probably specific for this infected cell model.

[Latent infection of continuous L23 cells caused by the agent of scrapie]. / Latentnaia infektsiia perevivaemykh kletok L23, vyzvannaia agentom skrepi.

Two models of latent infection of continuous L23 cells caused by scrapie agent were developed using the method of cell cloning as well as DEAE-dextran and lidase preparations. The infectious activity was found only in cell homogenate L-C and L-506 (cells of clone L23 latently infected with the Compton and C-506 strains of the scrapie agent). A direct relationship between the infectious activity of the cell homogenate and the intensity of brain lesions revealed by morphological examinations of brain sections from infected mice was observed. The cytoplasm of L-C and L-506 cells became strongly vacuolated, piknosis of the nuclei and delay in the cell growth rate were noted. Cytogenetic studies established changes in the model class of the experimental cultures as well as the appearance of rearranged chromosomes.

[Morphological, cytogenetic and proliferative characteristics of Syrian hamster HTC-2 and HTC-1 cells and of the HTCT tumor cell line transformed by herpes simplex type 2 virus]. / Morfologicheskie, tsitogeneticheskie i proliferativnye osobennosti transformirovannykh virusom prostogo gerpesa tipa 2 kletok siriiskogo khomiachka KhTK-2, KhTK-1 i linii opukholevykh kletok KhTKO.

Morphological, caryological, cytoproliferative, and isoenzyme studies of Syrian hamster cells HTC-2 and HTC-1 transformed by herpes simplex virus type 2 and a HTCT line of tumor cells were carried out. The hamster origin of these cell lines was established by chromosomal analysis and electrophoretic mobility of lactate dehydrogenase and glucose-6-phosphate-dehydrogenase isoenzymes. The transformed and tumor cell lines differ from normal cells by altered morphology, an aneuploid chromosome set and increased proliferative activity. Electron microscopic examination of transformed cells revealed increased amounts of filamentous and tubular structures in the cytoplasm. No retroviruses were found.

[Karyological characteristics of L cells and the identification with them, of the antigen of persisting Japanese encephalitis virus]. / Kariologicheskaia kharakteristika kletok L i identifikatsiia v nikh antigena persistiruiushchego virusa iaponskogo éntsefalita.

The immunofluorescence procedure revealed the virus-specific Japanese encephalitis virus antigen in latently infected L cells as well as differences in the percentage of antigen-positive cells and in the localization of the virus-specific antigen under conditions of acute, chromic, and latent infection and in the period of activation of the latent state of the virus at 28 degrees C. Karyological studied demonstrated no marked differences between persistently infected and original lines.