Модель биомедицинского клеточного продукта для доклинических исследований на крупном лабораторном животном

Objective: to develop a model of a biomedical cell product that is consistent with the «homologous drug» strategy  based on protocols for preparing the cell component and scaffold carrier for preclinical studies on a large laboratory  animal (pig). Materials and methods. Biomedical cell products and skin equivalents (SE), were formed using  plasma cryoprecipitate prepared from blood plasma of healthy donors and mesenchymal stem cells (MSCs) of  human adipose tissue. Cryoprecipitate from pig blood plasma and human adipose tissue-derived MSCs were used   to form model skin equivalents (mSE). Bright-field microscopy, phase-contrast microscopy (Leica DMI 3000B)  and fluorescence microscopy (Cytation 5 imager; BioTek, USA) were used to monitor the state of cells in the  culture and in the composition of the equivalents. Scaffolds for equivalents were tested for cytotoxicity (MTT test,  direct contact method). The cell distribution density was characterized by author’s method (Patent No. 2675376  of the Russian Federation). Results. An mSE was developed for preclinical studies on a large laboratory animal  (pig). In the mSE, components that change from halogen to xenogenic conditions during transplantation to the  animal were replaced. A comprehensive approach to preparing mSE was presented. It includes sampling of primary  pig biomaterial, extraction and characterization of adipose tissue-derived MSCs, preparation of a scaffold  carrier for the corresponding «homologous drug» strategy. Cytotoxicity of the mSE scaffold was evaluated. It  was shown that mSE provides mechanical support (similar to SE) to cells, as well as comparable development of  cellular events during cultivation. Conclusion. A model of a biomedical cell product was developed. This model  is consistent with the «homologous drug» strategy for preclinical studies on a large laboratory animal (pig). The  paper presented a comprehensive approach to developing a model equivalent based on protocols for preparation  and testing of the cellular component, the scaffold carrier and the ready-to-use model equivalent.Цель: разработать модель биомедицинского клеточного продукта, согласующуюся со стратегией «гомологичный препарат» на основе протоколов подготовки клеточной составляющей и скаффолда-носителя для доклинических исследований на крупном лабораторном животном (свинье). Материалы и методы. Биомедицинские клеточные продукты – эквиваленты кожи (ЭК) формировали с использованием криопреципитата плазмы крови здоровых доноров и мезенхимальных стволовых клеток (MSCs) жировой ткани человека. Для формирования модельных эквивалентов кожи (мЭК) использовали криопреципитат плазмы крови свиней и MSCs жировой ткани свиней. Наблюдение за состоянием клеток в культуре и в составе эквивалентов проводили с использованием методов светлого поля, фазового контраста (Leica DMI 3000B) и флуоресцентной микроскопии (имиджер Cytation 5; BioTek, USA). Скаффолды эквивалентов тестировали на цитотоксичность (МТТ-тест, метод прямого контакта). Характеристику плотности распределения клеток проводили авторским способом (Пат. № 2675376 РФ). Результаты. Разработан модельный эквивалент кожи (мЭК) для проведения доклинических исследований на крупном лабораторном животном (свинье). В мЭК замещены компоненты, переходящие из алогенных условий в ксеногенные при трансплантации животному. Представлен комплексный подход для подготовки мЭК, включающий забор первичного биоматериала свиньи, выделение и характеристику MSCs жировой ткани, подготовку скаффолда-носителя, соответствующего стратегии «гомологичный препарат». Проведена оценка цитотоксичности скаффолда мЭК. Показано, что мЭК обеспечивает аналогичную эквиваленту кожи (ЭК) механическую поддержку клеток и сопоставимое развитие клеточных событий при культивировании. Вывод. Разработана модель биомедицинского клеточного продукта, согласующаяся со стратегией «гомологичный препарат» для доклинических исследований на крупном лабораторном животном (свинье). Представлен комплексный подход, для разработки модельного эквивалента основанный на протоколах подготовки и тестирования клеточной составляющей, скаффолда-носителя и готового модельного эквивалента

Factors associated with diversity, quantity and zoonotic potential of ectoparasites on urban mice and voles

Wild rodents are important hosts for tick larvae but co-infestations with other mites and insects are largely neglected. Small rodents were trapped at four study sites in Berlin, Germany, to quantify their ectoparasite diversity. Host-specific, spatial and temporal occurrence of ectoparasites was determined to assess their influence on direct and indirect zoonotic risk due to mice and voles in an urban agglomeration. Rodent-associated arthropods were diverse, including 63 species observed on six host species with an overall prevalence of 99%. The tick Ixodes ricinus was the most prevalent species, found on 56% of the rodents. The trapping location clearly affected the presence of different rodent species and, therefore, the occurrence of particular host-specific parasites. In Berlin, fewer temporary and periodic parasite species as well as non-parasitic species (fleas, chiggers and nidicolous Gamasina) were detected than reported from rural areas. In addition, abundance of parasites with low host-specificity (ticks, fleas and chiggers) apparently decreased with increasing landscape fragmentation associated with a gradient of urbanisation. In contrast, stationary ectoparasites, closely adapted to the rodent host, such as the fur mites Myobiidae and Listrophoridae, were most abundant at the two urban sites. A direct zoonotic risk of infection for people may only be posed by Nosopsyllus fasciatus fleas, which were prevalent even in the city centre. More importantly, peridomestic rodents clearly supported the life cycle of ticks in the city as hosts for their subadult stages. In addition to trapping location, season, host species, body condition and host sex, infestation with fleas, gamasid Laelapidae mites and prostigmatic Myobiidae mites were associated with significantly altered abundance of I. ricinus larvae on mice and voles. Whether this is caused by predation, grooming behaviour or interaction with the host immune system is unclear. The present study constitutes a basis to identify interactions and vector function of rodent-associated arthropods and their potential impact on zoonotic diseases

ОСОБЕННОСТИ СУБПОПУЛЯЦИОННОГО СОСТАВА ЛИМФОЦИТОВ У ДЕТЕЙ ПРИ АТОПИЧЕСКОМ ДЕРМАТИТЕ

Article is devoted the analysis of subpopulation structure of Tlymphotsytes at different stages of an atopic dermatitis at children.Статья посвящена анализу субпопуляционного состава Т - лимфоцитов при разных стадиях атопического дерматита у детей

Biomedical cell product model for preclinical studies carried out on a large laboratory animal

Objective: to develop a model of a biomedical cell product that is consistent with the «homologous drug» strategy  based on protocols for preparing the cell component and scaffold carrier for preclinical studies on a large laboratory  animal (pig). Materials and methods. Biomedical cell products and skin equivalents (SE), were formed using  plasma cryoprecipitate prepared from blood plasma of healthy donors and mesenchymal stem cells (MSCs) of  human adipose tissue. Cryoprecipitate from pig blood plasma and human adipose tissue-derived MSCs were used   to form model skin equivalents (mSE). Bright-field microscopy, phase-contrast microscopy (Leica DMI 3000B)  and fluorescence microscopy (Cytation 5 imager; BioTek, USA) were used to monitor the state of cells in the  culture and in the composition of the equivalents. Scaffolds for equivalents were tested for cytotoxicity (MTT test,  direct contact method). The cell distribution density was characterized by author’s method (Patent No. 2675376  of the Russian Federation). Results. An mSE was developed for preclinical studies on a large laboratory animal  (pig). In the mSE, components that change from halogen to xenogenic conditions during transplantation to the  animal were replaced. A comprehensive approach to preparing mSE was presented. It includes sampling of primary  pig biomaterial, extraction and characterization of adipose tissue-derived MSCs, preparation of a scaffold  carrier for the corresponding «homologous drug» strategy. Cytotoxicity of the mSE scaffold was evaluated. It  was shown that mSE provides mechanical support (similar to SE) to cells, as well as comparable development of  cellular events during cultivation. Conclusion. A model of a biomedical cell product was developed. This model  is consistent with the «homologous drug» strategy for preclinical studies on a large laboratory animal (pig). The  paper presented a comprehensive approach to developing a model equivalent based on protocols for preparation  and testing of the cellular component, the scaffold carrier and the ready-to-use model equivalent

CYTOKINES AND HISTOPATHOLOGIC PATTERN OF MALIGNANT NEOPLASMS IN GASTROINTESTINAL CANCER

The patients with gastrointestinal cancer exhibit high contents of TNFα, IL-1β, IL-2, IL-6, IL-8, IFNγ, IL-1Ra and increased levels of anti-IFNα antibodies. Significant correlations are detected between histopathological parameters of the tumors, and the levels of TNFα, IL-2, IL-6, IL-8, IFNα, IFNγ, as well as antibodies against TNFα и IFNα. Moreover, we have determined diagnostically significant levels of cytokines and their combinations. On the basis of estimated diagnostic values, one may evaluate both current status of the tumor, as well as its potential malignancy grade

CYTOKINE-PRODUCING FUNCTION OF PERIPHERAL BLOOD CELLS IN COLORECTAL CANCER PATIENTS

Аbstract. The aim of present study was to investigate an influence of polyclonal activators upon cytokine-producing function of peripheral blood cells in the patients with colorectal adenocarcinomas. It was revealed that the stimulated production of IL-1β, IL-1ra, IL-2, IL-17, IL-18 and IFNγ by whole blood cells treated with polyclonal activators was significantly decreased in colorectal cancer patients, as compared to healthy individuals. The stimulated IL-1ra and IL-17 production was depressed during the tumor progression, along with increased release of TNFα and IL-8. It was found that the stimulation index of polyclonal activators upon production of IL-17 by whole blood cells (i.e., a cytokine production ratio of stimulated versus resting cells) may be used as a reliable pre-surgery predictor of tumor invasion, the latter being an important histopathological parameter for clinical oncologists. (Med. Immunol., 2011, vol. 13, N 2-3, pp 197-20

Biosphere: Analysis of Value Parameters [Электронный ресурс]

Value parameters of biosphere in the view of its qualitative-quantitative characteristics are analyzed. Interpretation variant of value estimation of deformation processes of organic and inorganic nature under the impact of anthropogenic activity from theИспользуемые программы: Adobe Acrobat.Труды сотрудников СГАУ (электрон. версия

THE INFLUENCE OF HUMAN LEUKEMIA DIFFERENTIATION FACTOR ON ABILITY OF BLOOD CELLS TO PRODUCE IL-1β, IL-1ra AND IL-8 IN CHRONIC ATROPHIC GASTRITIS, ADENOMAS AND ADENOCARCINOMAS OF THE STOMACH

We studied biological effects of Human Leukemia Differentiation Factor (HLDF), first derived from the cultural media of human myelogenous HL-60 leukemia cells, on the production of IL-1β, IL-1ra and IL-8 by immune cells from the patients with chronic atrophic gastritis, adenomas and adenocarcinomas of the stomach. To evaluate the influence of HLDF on cytokine-producing function of the whole blood cells, the latter were incubated with this factor or without it. The levels of IL-1β, IL-1ra and IL-8 were determined in supernates of these cells. The stimulation indices of HLDF upon cytokine production were estimated as a cytokine production ratio of stimulated versus resting cells. Histological analysis of gastric mucous tissue samples and removed tumors was also performed. It was revealed that, in chronic atrophic gastritis, HLDF significantly increased IL-1ra and IL-8production. In patients with stomach adenomas and adenocarcinomas, HLDF increased IL-1ra, IL-8 and IL-1β production. In cases of stomach adenomas, the indices of HLDF stimulation upon blood cells cytokine production are significant higher than in patients with chronic atrophic gastritis. Interdependence study between the stimulation indices of HLDF and histological parameters of gastric mucosa and tumor samples have shown a negative correlations between the stimulation index of HLDF upon IL-1ra production and the grade of intestinal metaplasia and dysplasia of the adenoma epithelium, and a positive correlation between the stimulation index of HLDF on IL-8 production and the intensity of lymphoid infiltration of the adenomas. In patients with gastric adenocarcinomas, a positive correlation between the stimulation index of HLDF on IL-8 production and number of blood vessels with tumor embolus was revealed. One may presume that immune cells activation caused by HLDF may exert following actions: (1) triggering some processes of chronic inflammation that underly malignancy development, and (2) the mechanisms restricting the disturbances of epithelial regeneration. Finally, the results of HLDF effects depend on balance between proand antioncogenic cytokines produced by immune cells