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PL
Wyselekcjonowano szczepy Staphylococcus aureus, które wykorzystano do hodowli in vitro z dodatkiem sub-MIC wybranych antybiotyków oraz oceny podatności na fagocytozę i zabijanie przez granulocyty królicze. Wykazano, że podatność większości szczepów S. aureus na fagocytozę i zabijanie przez granulocyty królicze zwiększa się w sposób istotny po wcześniejszym ich kontakcie z sub-MIC badanych antybiotyków.
EN
The aim of the study was to assess the of choosen antibiotics in subinhibitory concentrations (sub-MIC) on the sensitive of Staphylococcus aureus cells to phagocytosis and killing by rabbit granulocytes. The following antibiotics were used: cloxacillin, cefadroxil, cefuroxim, cefotaxim, gentamicin, netilmicin, lincomicin, doxycycline and riphamicin. A total of 144 S. aureus strains with varied sensitivity to these antibiotics were selected for the study. The experiment used granulocytes isolated from rabbit blood and S. aureus strains incubated for 18 h in TSB broth containing antibiotics in the concentrations of 0.1 MIC, 0.2 MIC and 0.5 MIC, and in the antibiotics-free medium. Phagocytosis was assessed by the method of differential staining with acridine orange and crystal violet, allowing simultaneous determination of phagocytised and killed S. aureus cell counts. The findings revealed that the culture of S. aureus in the presence of all the antibiotics used in subinhibitory concentrations increased significantly the susceptibility of most S. aureus strains to phagocytosis and killing by granulocytes. The above effect usually occurred in the concetrations of 0.1 MIC (54.2%), more seldom in 0.2 MIC (13%) and 0.5 MIC (15% of strains). Each group of S.aureus contained some which showed no change in susceptibility following culture with the chemotherapeutic agents in subinhibitory concentrations (26.3%). Insensitive strains to the subinhibitory effects were equally common among susceptible (27%), intermediate (23%) and resistant (26%) strains of S. aureus to the antibiotics used. No statistically significant reduction was noted in phagocytosis or killing by rabbit granulocytes. No correlation was observed between the susceptibility to the subinhibitory effects of the antibiotics involved and their biochemical mechanisms.
PL
Zbadano adhezję ludzkich granulocytów i limfocytów T do komórek śródbłonka naczyniowego linii HMEC-1 stymulowanych za pomocą endotoksyny (LPS) i enterotoksyny (BFT) B. fragilis. Stwierdzono przyleganie granulocytów oraz limfocytów T spoczynkowych i aktywowanych PMA do komórek endotelialnych po stymulacji endotoksyną i enterotoksyną B. fragilis. Aktywność obu toksyn B. fragilis w procesie stymulacji adhezji leukocytów do śródbłonka naczyniowego jest mniejsza od aktywności LPS E. coli O55:B5.
EN
The aim of presented study was to estimates the number of human granulocytes and T lymphocytes adhering to 1 mm² of vascular endothelial cell culture stimulated by Bacteroides fragilis endotoxins (LPS) and enterotoxin (BFT). HMEC-1 cells were activated with bacterial preparations at the concentration of 10 (µg/ml for 4 and 24 hours. Granulocytes and T lymphocytes were isolated from peripheral blood of healthy blood donors. The adhesion tests of granulocytes and adhesion tests of resting and activated with PMA (at the concentration of 10 ng/ml) T lymphocytes to the non-stimulated and stimulated by B fragilis compounds (LPS and BFT) vascular endothelium were performed. The number of viable leukocytes, which adhered to the endothelium, was determined using inverted microscope (magnification 200 x). The results were presented as the number of viable cells adhering to 1 mm² of the endothelial cell culture. The results of experiments indicate that granulocytes and T lymphocytes (resting and after activation with PMA even in greater number) adhere to the endothelial cells stimulated by B. fragilis endotoxins and enterotoxin. B. fragilis toxins are weaker stimulants of human leukocyte adhesion to the HMEC-1 cells than E. coli O55:B5 LPS. B. fragilis LPS and BFT preparations stimulate endothelial cells to the adhesion of granulocytes in similar manner, whereas the activation of vascular endothelium to the adhesion of T lymphocytes is differentiated.
PL
Przeprowadzono testy adhezji ludzkich limfocytów T i granulocytów do komórek śródbłonka naczyniowego linii HMEC-1 stymulowanych przy użyciu polisacharydu otoczkowego (CPS), lipopolisacharydów (LPS) oraz komponentów wielocukrowych (PS) i lipidowych (lipid A) LPS. Stwierdzono, że limfocyty T spoczynkowe i aktywowane PMA oraz granulocyty przylegają do komórek endotelialnych stymulowanych za pomocą antygenów powierzchniowych B. thetaiotaomicron. Aktywność w tym procesie preparatów LPS i CPS B. thetaioitaomicron jest mniejsza od aktywności LPS E. coli 055:B5.
EN
The aim of this study was to assay the degree of human T lymphocyte and granulocyte adhesion to the vascular endothelial cells stimulated by Bacteroides thetaiotaomicron lipopolysaccharides, components of LPS and capsular polysaccharide. HMEC-1 cells were activated with bacterial preparations in concentration 10 ug/ml for 4 and 24 hours. T lymphocytes and granulocytes were isolated from peripheral blood of healthy blood donors. Thereafter, the adhesion tests of granulocytes and adhesion tests of non-activated and activated with PMA ( in concentration 10 ng/ml) T lymphocytes to the resting and stimulated vascular endothelium were performed. The number of viable cells, which adhered to the endothelium, was determined using inverted microscope (magnification 200x). The results were presented as the number of viable cells adhering to 1 mm2 of the endothelial cell culture. The obtained results indicate that granulocytes and T lymphocytes (resting and activated with PMA) adhere to the endothelial cells stimulated by B. thetaiotaomicron cell-surface antigens. B. thetaiotaomicron lipopolysaccharides and capsular polysaccharide are weaker stimulants of human leukocyte adhesion to the HMEC-1 cells than E. coli 055:B5 LPS.
PL
Oleanozydy z aralii mandżurskiej (OAM) należą do polisacharydów. Ponadto aralia mandżurska podobnie jak żeń-szeń (Panax ginseng) należy do rodziny Araliaceae. Wyciągi z Panax ginseng są immunostymulatorami swoistej odpowiedzi komórkowej i humoralnej. Celem pracy była ocena OAM podawanego doustnie na kształtowanie się parametrów fagocytozy granulocytów pochodzenia zwierzęcego. Stwierdzono stabilizację leukocytozy, poziomów granulocytów oraz wzmożenie aktywności fagocytarnej granulocytów - pochłaniania i trawienia.
EN
Oleanosides from Aralia mandshurica Rupr. et Maxim (OMA), are polysaccharides. In a similar way as gingseng (Panax gingseng) Manchurian aralia belongs to Araliaceae family. Extracts from Panax ginseng are known to be immunostimulators of specific cellular and humoral response. The aim of the study was to evaluate the effect of OMA, administered orally, exerted on the formation of parameters for phagocytosis of granulocytes being of animal origin. It was disclosed stabilization of leucocytosis, granulocytes and statistically significant intensification of phagocytic activity of granulocytes - ingestion and digestion.
EN
The aim of the study was to evaluate the activity of peripheral blood granulocytes in rabbits with chronic trichophytosis by using commercial sets of Phagotest and Burstest adopted to flow cytometry. A strong suppression of unspecific cellular immune responses was found in rabbits with chronic trichophytosis. Once fungal infection is established it can only be destroyed by T cell-mediated mechanisms. T cells primarily function by activating macrophages and by promoting epidermal growth and keratinization. The destroyed unspecific immune mechanisms are manifested by decreased phagocyte activity and oxygen metabolism of granulocytes. Therefore, it is reasonable to assume that when vaccine therapy is used to treat chronic trichophytosis in rabbits with unspecific immunity it should be modulated in order to restore the destroyed mechanisms of unspecific immunity. The latter's normal functioning is a prerequisite for the development of specific antifungal immunity.
PL
Zbadano produkcję tlenku azotu (NO) i reaktywnych form tlenu przez granulocyty i monocyty zakażane in vitro żywymi bakteriami BCG, pochodzące od zdrowych dawców krwi. Zaobserwowano znaczące różnice w reakcjach biochemicznych stymulowanych prątkami BCG, zachodzące w badanych komórkach. Równocześnie oceniano aktywność fagocytów w hodowlach prowadzonych w obecności cytokin.
EN
The production of nitric oxide (NO) and reactive oxygen intermediates in granulocytes and macrophages from healthy volunteers, infected in vitro with live Bacille Calmette-Guerin (BCG) mycobacteria, was estimated. Significant differences in the biochemical reactions induced by BCG bacilli in granulocytes and monocytes are described. The activity of phagocytes was also investigated in the cultures with cytokines: IFN-y, TNF-a, GM-CSF, IL-4.
PL
IL-3 najeży do multiliniowych stymulatorów hematopoezy, działających w obrębie puli komórek macierzystych. Istnieją nieliczne doniesienia, iż IL-3 podobnie jak inne cytokiny hematopoetyczne działa na funkcje dojrzałych komórek, ale stwierdzono to tylko w warunkach in vitro. W niniejszej pracy wykazano wpływ IL-3 na funkcje fagocytarne, właściwości bakteriobójcze i aktywność enzymów lizosomalnych granulocytów w warunkach żywego organizmu myszy.
EN
Interleukin-3 is a muItipotential hematopoietic growth factor, which like other colony stimulating factors (CSFs) is effective „in vitro" stimulation of the mature cells function. It was found that IL-3 synergistically with GM-CSF and G-CSF stimulated the proliferation of the granulocytes. Therefore the purpose of this investigation was the evaluation „in vivo" of the influences of IL-3 on the phagocytosis, bactericidal activity, and enzyme activities of granulocytes. IL-3 injected into mice subcutaneously during 5 days in dose 1 µg/kg/d. The examination of percent of cells phagocytising bacteria (Staphylococcus aureus), NBT test and bactericidal activity were performed every day and evident increase of the tested parameters was found. Additionally the enzyme activities in primary granules were measured and showed on increase of acid phosphatase and peroxidase activity.
PL
Stwierdzono, że granulocyty ludzkiej krwi obwodowej w warunkach działania dichlorfosu (pestycydu fosforoorganicznego) in vitro wykazywały istotnie zwiększoną aktywność fagocytarną (wzrost indeksu fagocytarnego) przy niezmienionej lub obniżonej (w porównaniu do kontroli) zdolności do redukcji NBT, wyrażającej poziom aktywności bakteriobójczej tych komórek. Wyniki wstępnych badań aktywności fosfatazy alkalicznej w granulocytach wskazują na zależne od dawki dichlorfosu obniżenie aktywności tego enzymu w porównaniu z kontrolą.
EN
The purpose of the study was to assess the phagocytic activity and bactericidal function of human peripheral blood neutrophils exposed to dichlorfos (DDVP) an organic phosphorus pesticide in vitro. It was found that the values of the phagocytosis index in neutrophil cultures incubated with DDVP in doses of 70 and 100 µM increased with increasing pesticide concentration and depended on incubation duration (1 h and 2 h). The proportion of non-phagocytizing neutrophils remained at a level similar to that in controls. The value of NBT reduction in neutrophils (indicating their bactericidal activity) after 1 hour of incubation with DDVP remained at the level of control values, but after 2 hours it decreased - significantly at DDVP concentration of 70 µМ, and at DDVP concentration 100 µM p=0.05. These results show that DDVP causes in neutrophils a selective stimulation of one step of phagocytosis connected with engulfing of particles, with simultaneous inhibition and/or damage to the mechanisms connected with aerobic killing of microorganisms.
EN
The aim of the study was to evaluate the CD3, CD4, CD8, CD19, CD25 T lymphocyte subpopulations and the state of granulocytes activity in the peripheral blood of rabbits infected with Trichophyton mentagrophytes and immunized against trichophytosis by using the flow cytometry method. Our study revealed significant suppression of non-specific cellular antimycotic immunity in rabbits during the development of fungal lesions, which manifested a significant decrease in the phagocytic activity and oxygen metabolism of granulocytes and the decrease of CD3, CD4 subpopulations and CD25 T lymphocytes. Simultaneously, the increase of suppressor CD8 T cells and CD4/CD8 T-cells ratio were observed. The vaccine Alopevac can be used as an effective vaccine against rabbit trichophytosis. Alopevac restored non-specific cellular antimycotic immunity and proper CD4/CD8 T-cells ratio, which contributed to the effective elimination of fungal lesions.
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