Enzyme replacement therapy (ERT) is one of the most efficient treatments for lysosomal storage diseases. Type 1 Gaucher disease is caused by β-glucocerebrosidase enzyme deficiency, which may be compensated for by intravenous infusions of imiglucerase—a recombinant enzyme. Imiglucerase targets macrophages and enters these cells via interaction with mannose receptors on the cell membrane. Characterisation of internalization of enzymes by target cells is important in the context of the development of new medicines and production of existing ERT medicines. The peritoneal and alveolar macrophages, as well as macrophages of the spleen of small laboratory animals (rats and mice) are widely used in such studies. However, isolation of cells from animal sources raises ethical issues, and therefore continuous mammalian cell lines may offer an attractive alternative. The aim of the study: to conduct comparative studies on the internalization of recombinant imiglucerase into mouse peritoneal macrophages and L929 mouse fibroblasts. Materials and methods: CerezymeR batches 7HV0913, C6214H05, 7HV0888 (Genzyme Ltd., UK); Glurazim batches 020416, 011117, 021117 (LLC “IBC “Generium”, Russia). We used peritoneal macrophages obtained from BALB/c mice and L929 mouse fibroblasts. The cells were cultured in DMEM/F12 complete growth medium with 10% fetal bovine serum. The activity of imiglucerase internalized into the cells was evaluated spectrophotometrically by hydrolysis of the artificial substrate—4-methylumbelliferyl-β-Dglucopyranoside. Results: the study compared internalization of recombinant imiglucerase (the active ingredient of CerezymeR and Glurazim) by mouse peritoneal macrophages and L929 mouse fibroblasts. It was demonstrated that the medicines activity in the lysates of peritoneal macrophages is comparable with that in the lysates of L929 mouse fibroblasts. Regardless of the model system, the activity of Glurazim stayed within the acceptable range (80–125%) established for biosimilar products. Conclusions: the experiments proved that L929 mouse fibroblasts could be recommended for assessment of internalization of recombinant imiglucerase.
Irrational placement of crops without taking into account the “genotype-environment” interaction leads to significant economic losses, especially in conditions of climate change in the manifestation of thermal stress-factors. The purpose of the work is to ensure the stability of the perennial crops fruiting from the position of maximum realization of their hereditary basis in various growing conditions. Using the methods of genetic-physiological, biological, genetic-physiological, meteorological data and digital technologies, a new approach was proposed for managing crop productivity on the example of fruit crops varieties. The importance of the development increases dramatically due to the fact that its use provides stabilization of the fruiting of perennial crops varieties in specific areas of cultivation, taking into account climate change. Using research results will reduce the risk of yield loss from environmental stress-factors. On the basis of a long-term analysis (1986-2018), the probability of manifestation of the temperatures of the winter-spring period, which are destructive for apricot plants, has been studied in different zones of the Krasnodar Territory. The critical minimum temperatures for the fruiting of apricot genotypes of different frost resistance of Krasnoshchekiy (lowered) and 1-22 hybrid (increased), breeding of North Caucasus Federal scientific center of horticulture, viticulture, winemaking for the phases of their development were determined. The manifestations of the minimum temperatures of the winter-spring period were estimated for 30 meteorological stations of the Krasnodar Territory based on data from an analysis of exceeding their critical values in the development phases of the studied varieties. Their response (also in the context of phenological phases) to the temporal and spatial dynamics of limiting factors of the environment is analyzed. The obtained data for each meteorological station is imported into the geographic information system ILWIS, and interpolated to obtain the results of rational distribution of the studied varieties of apricot in the form of digital maps. The constructed digital maps of rational distribution of specific varieties by development phases reveal the adaptive potential of varieties in the conditions of climate change. A new approach to managing the productivity of fruit crops varieties on the example of apricot in various zones of the Krasnodar Territory is proposed.
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