RESUMEN
The leading pathological mechanisms of Alzheimer's disease are amyloidosis and inflammation. The presented work was aimed to study the effect of human peripheral blood mononuclear cells (hPBMcs) cells-matrix adhesion on their pro-inflammatory state in vitro. Although direct interaction of Ðß42 to PBMC is not a cellular model of Alzheimer's disease, PBMCs may serve as test cells to detect Ðß42-dependent molecular effects in monitoring disease progression. Peripheral blood mononuclear cells (PBMCs) are used to assess changes in cytokines released in response to diseases or Alzheimer's disease-specific cytotoxic molecules such as Aß42. The effect of recombinant amyloid ß-peptide rÐß42 on the concentration of endogenous amyloid ß-peptide Aß40 and pro-inflammatory cytokines TNFα and IL-1ß in human peripheral blood mononuclear cells that were cultured in suspension and immobilized in alginate microcarriers for 24 h were investigated. The localization and accumulation of Aß40 and rAß42 peptides in cells, as well as quantitative determination of the concentration of Aß40 peptide, TNFα and IL-1ß cytokines, was performed by intravital fluorescence imaging. The results were qualitatively similar for both cell models. It was determined that the content of TNFα and Aß40 in the absence of rAß42 in the incubation medium did not change for 24 h after incubation, and the content of IL-1ß was lower compared to the cells that were not incubated. Incubation of cells in vitro with exogenous rAß42 led to an increase in the intracellular content of TNFα and Aß40, and no accumulation of IL-1ß in cells was observed. The accumulation of Aß40 in the cytoplasm was accompanied by the aggregation of rAß42 on the outer surface of the cell plasma membrane. It was shown that the basic levels of indicators and the intensity of the response of immobilized cells to an exogenous stimulus were significantly greater than those of cells in suspension. To explore whether non-neuronal cells effects in alginate microcarriers were cell-matrix adhesion mediated, we tested the effect of blocking ß1 integrins on proamyloidogenic and proinflammation cellular state. Immobilization within alginate hydrogels after incubation with the ß1 integrins blocking antibodies showed a remarkable inhibition of TNFα and Aß40 accumulation in rAß42-treated cells. It can be concluded that activation of signal transduction and synthesizing activity of a portion of mononuclear cells of human peripheral blood is possible (can significantly increase) in the presence of cell-matrix adhesion.
RESUMEN
Butanol-producing strains Clostridium sp. UCM B-7570 and C. acetobutylicum UCM B-7407 were used for research from "Collection of strains of microorganisms and plant lines for food and agricultural biotechnology" of the Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine, glycerol (BASF, Germany) and apple pomace (total moisture 4%) after apple juice production. The aim of this work was to study the possibility of using apple pomace by domestic butanol-producing strains of Clostridium sp. UCM B-7570 and C. acetobutylicum UCM B-7407 as a substrate. Producers were cultured on medium with different concentrations of apple pomace, glycerol was used for the inoculation. The presence of ethanol, acetone, and butanol in the culture liquid was determined using a gas chromatograph. It was determined that a significant part of the macrocomponent composition of the extracts can be used in bioconversion by producing strains of the genus Clostridium. It was determined that the highest concentration of butanol (10 g/dm3) was at a concentration of 120 g/dm3 in the extracts. The obtained data showed the possibility of using apple pomace as a substrate in biobutanol technology.
RESUMEN
The rapid development of new molecular methods and approaches, sequencing technologies, has provided new insights into genetic and structural features of bacterial genomes. Information about the genetic organization of metabolic pathways and their regulatory elements has greatly contributed to the increase in the number of studies related to the construction of new bacterial strains with improved characteristics. In this study, the entire genome of the producing strain Clostridium sp. UCM Ð-7570 from the "Collection of producing strains of microorganisms and plant lines for food and agricultural biotechnology" of Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine was sequenced and characterized. The genome was assembled into the scaffold with a total size of 4,470,321 bp and a GC content of 29.7%. The total number of genes identified was 4262, of which 4057 encoded proteins, 10 were rRNA operons, and 80 were tRNA genes. The genes of the sequenced genome encoding enzymes involved in butanol fermentation were found and analyzed. They were organized into cluster structures, and their protein sequences were found to be similar to the corresponding strains of C. acetobutylicum, C. beijerinckii, and C. pasteurianum type strains with the highest similarity to the latter. Thus, Clostridium sp. UCM Ð-7570 producing strain was identified as C. pasteurianum and suggested for metabolic engineering purposes.