Perspectives on the Use of Small Noncoding RNAs as a Therapy for Severe Virus-Induced Disease Manifestations and Late Complications.

Many viruses appear each year. Some of these viruses result in severe disease and even death. The frequency of epidemics and pandemics is growing at an alarming rate. The lack of virus-specific etiopathogenic drugs necessitates the search for new tools for the complex treatment of severe viral diseases and their late complications. Small noncoding RNAs and their antagonists may be effective therapeutic tools for preventing virus-induced damage to targeted epithelial cells and surrounding tissues in the manifestation stage. Moreover, sncRNAs could interfere with the virus-interacting host genes that trigger the malignant transformation of target cells as a late complication of severe viral diseases.

The full recovery of mice (Mus Musculus C57BL/6 strain) from virus-induced sarcoma after treatment with a complex of DDMC delivery system and sncRNAs.

BACKGROUND: Virus-induced cellular genetic modifications result in the development of many human cancers. METHODS: In our experiments, we used the RVP3 cell line, which produce primary mouse virus-induced sarcoma in 100% of cases. Inbreed 4-week-old female C57BL/6 mice were injected subcutaneously in the interscapular region with RVP3 cells. Three groups of mice were used. For treatment, one and/or two intravenous injections of a complex of small non-coding RNAs (sncRNAs) a-miR-155, piR-30074, and miR-125b with a 2-diethylaminoethyl-dextran methyl methacrylate copolymer (DDMC) delivery system were used. The first group consisted of untreated animals (control). The second group was treated with one injection of complex DDMC/sncRNAs (1st group). The third group was treated with two injections of complex DDMC/sncRNAs (2nd group). The tumors were removed aseptically, freed of necrotic material, and used with spleen and lungs for subsequent RT-PCR and immunofluorescence experiments, or stained with Leishman-Romanowski dye. RESULTS: As a result, the mice fully recovered from virus-induced sarcoma after two treatments with a complex including the DDMC vector and a-miR-155, piR-30074, and miR-125b. In vitro studies showed genetic and morphological transformations of murine cancer cells after the injections. CONCLUSIONS: Treatment of virus-induced sarcoma of mice with a-miR-155, piR-30074, and miR-125b as active component of anti-cancer complex and DDMC vector as delivery system due to epigenetic-regulated transformation of cancer cells into cells with non-cancerous physiology and morphology and full recovery of disease.

Reprogramming of CaCo2 colorectal cancer cells after using the complex of poly-(N-vinylpyrrolidone) with small non-coding RNAs.

Small non-coding RNAs control normal development and differentiation in the embryo. These regulatory molecules play a key role in the development of human diseases and are used often today for researching new treatments for different pathologies. In this study, CaCo2 colorectal adenocarcinoma cells were initially epigenetically reprogrammed and transformed into CD4+ cells with nano-sized complexes of amphiphilic poly-(N-vinylpyrrolidone) (PVP) with miRNA-152 and piRNA-30074. The transformation of cells was confirmed by morphological and genetic changes in the dynamic of reprogramming. CD4+ lymphocytes marker was detected using immunofluorescence. Amphiphilic poly-(N-vinylpyrrolidone)/small non-coding RNAs complexes were investigated for transfection efficiency and duration of transfection of CaCo2 colorectal adenocarcinoma cells using fluorescence.

Small non-coding RNAs as regulators of structural evolution and carcinogenesis.

Small non-coding RNAs (sncRNAs) are part of non-coding oligonucleotide regulators with wide physiologic and morphologic functions. They control genetic programing of cells, and may modulate processes of differentiation and death. Biogenesis of sncRNAs is now known, and some sncRNAs have been proposed as markers of malignization. Epigenetic therapy is based on the use of newly discovered genetic modifiers, such as sncRNAs, micro-RNAs, and theirs mimics. However, role of sncRNAs in structural evolution and mechanisms of adaptation is not clearly understood. Certainly, non-coding RNAs participate in processes of cellular and organismal adaptation as well as cellular and tissue structural transformation as response to changing of environmental neighbouring. Investigations into these functions of sncRNAs may be the basis of future epigenetic environmental medicine.

Effects of high tidal volume mechanical ventilation on production of cytokines, iNOS, and MIP-1ß proteins in pigs.

The aim of this study was to investigate longitudinal changes of the pulmonary inflammatory process as a result of mechanical stress due to mechanical ventilation. The concentrations of IL-8, TNF-α, MIP-1ß, nitrites/nitrates, and inducible nitric oxide synthases (iNOS) were investigated indicate in bronchoalveolar lavage (BAL). Twenty-three piglets were divided into three groups. Group I: animals breathing spontaneously; group II: mechanical ventilation (tidal volume (TV) = 7 mL/kg, PEEP = 5 cmH(2)O); group III: mechanical ventilation (TV = 15 mL/kg, PEEP = 0 cmH(2)0). Concentrations of BAL nitrites/nitrates from groups II and III increased during the first hour of mechanical ventilation (P = .03 and .02, respectively). The highest expression of iNOS was observed during the first hour in groups II and III. IL-8 concentration increased significantly in groups II and III. Production of TNF-α increased significantly in group III during the second and third hour (P = .01). Concentration of MIP-1ß was significantly increased in groups II and III after the first hour (P = .012 and P = .008, respectively).

Regulation of immune responses, apoptosis, and tumorigenesis by separate FOXP-3-dependent genes: connection with clinical manifestations.

Recently, forkhead/winged-helix family box protein P3 (FOXP-3) was described as the main regulator of regulatory T cells' activity. This transcription factor has the ability to control the immunosuppressive response of regulatory T cells. FOXP-3 has binding sites for different genes specific for proteins with various important functions. In this article, selected FOXP-3-dependent genes with known functions were divided into two groups. The first group of genes has main immunoregulatory functions, and the second group has the ability to regulate apoptosis and tumorigenesis. Investigation of the functions of all FOXP-3-dependent genes opens perspectives for applications in different fields of basic and clinical research.

Influence of the lung mechanical ventilation with injurious parameters on 7-ketocholesterol synthesis in Sus Scrofa.

The aim of work was to investigate changes of 7-ketocholesterol synthesis in alveolar macrophages in the dynamic of lung mechanical ventilation with injurious parameters. The goal of in vitro part of work was to observe influence of 7-ketocholesterol on iNOS and MIP1 Beta production in bronchoalveolar lavage fluid (BALF) cells. We used 17 healthy domestic pigs randomly assigned into two treatment groups: group I with mechanical ventilation with physiological parameters; group || underwent injurious ventilation with high volume tidal (VT) and low positive end expiratory pressure (PEEP). ells were analyzed for CYP27A1 protein and gene expression levels, 7-ketocholesterol production. In alveolar macrophages of group ||, we obtained increase of production of CYP27A1 protein and 7-ketocholesterol, as well as the expression of the CYP27A1 gene at the 2nd hour of ventilation. In the in vitro experiments we show dose-dependent increase of MIP1 Beta and decrease of CYP27A1, iNOS protein production after 7-ketocholesterol treatment.