Anti-FITC polyclonal antibody [HRP]

Current study performed to estimate the phagocytic activity of blood monocytes of varying phenotypes exposed to MRSA and MSSA strains. Objects: Blood monocytes were collected from 25 healthy adults (age: 25- 45 years). Live suspensions of MRSA/MSSA strains were used at concentration of 106 colony-forming units (CFU)/mL. Metods. Phagocytic functions were estimated by using fluorescein isothiocyanate ( FITC)-labelled MRSA and MSSA strains followed by running flow cytometry on FC 500 series flow cytometer (Beckman Coulter, USA). Whole peripheral blood cells were directly labelled with immunofluorescently tagged monoclonal CD14-PE/CD45-ECD/HLA-DR-PC5/CD16-PC7 antibodies (Beckman Coulter, USA). Respiratory burst intensity was evaluated in monocytes by measuring activity of lucigenin- and luminol-dependent spontaneous and induced chemiluminescence. Monocytes were induced by using live suspension of MRSA/MSSA strains at a concentration of 106 CFU/mL. Results and discussion. While studying luminol-dependent monocyte activities after exposure to MRSA vs. MSSA, it was observed a 3.5-fold decreased curve square, whereas lucigenin-dependent chemiluminescence was increased by 6-fold. Compared to MSSA exposure, index of activation (IA) was decreased by 1.1-fold in response to MRSA exposure that was confirmed by lowered release of reactive oxygen species (ROS) from monocytes in response to MRSA exposure. Moreover, IRSS increased by 1.3-fold upon MRSA exposure. Examining monocyte oxygen-independent phagocytosis against MRSA vs. MSSA revealed significantly increased phagocytic number and concomitantly decreased phagocytic index. An evaluation of the activities of various monocyte subsets in response to MRSA vs. MSSA revealed increased phagocytic index by 1.5-fold for CD14(low)CD16(+) and CD14(+)CD16(+) monocyte subsets as well as 3-fold for CD14(+)CD16(-) monocytes. Counts for all phagocytic subsets were decreased (1.4-, 1.5- and 4-fold for CD14(low)CD16(+), CD14(+)CD16(+) and CD14(+) CD16(-) monocytes, respectively). To summarize, intensity of the respiratory burst was lowered upon MRSA exposure and percentage of monocyte subsets. Overall deficiency of superoxide anion production was observed in response to MRSA. In contrast, oxygen-independent event revealed phenotypic changes in frequency of peripheral blood monocytes upon MRSA exposure. We observed that CD14(+) CD16(-) classical monocytes were more rapidly activated. Conclusion. Thus, we concluded that CD14(+)CD16(-) monocytes became more rapidly activated but exhibited less effective phagocytosis, whereas CD14(+)CD16(+) and CD14(low)CD16(+) monocytes were more slowly activated and demonstrated stronger phagocytic activity.

This study examines the effect of L-carnitine (LC) on chilled ram semen stored for up to 96 hr. Semen samples were collected, placed in a skimmed milk + 6% egg yolk extender, pooled, aliquoted and diluted with the same extender supplemented with different LC concentration: 0 (control), 1 mM (LC1), 2.5 mM (LC2.5), 5 mM (LC5), 7.5 mM (LC7.5) or10 mM (LC10). Sperm kinetics and membranes (plasma, acrosome and mitochondrial) were examined using the CASA system and triple fluorescence staining (PI/ PNA-FITC/Mitotracker). The progressive motility was greater (p < .05) with LC7.5 treatment than the control sperm at 96 hr. The curvilinear velocity (p < .01) and the percentage of sperm with intact membranes (plasma/acrosome/mitochondria) (p .01) were greater with all LC treatments than the control group at all times. Straight line velocity was greater (p .01) with LC5 and LC7.5 treatments than the control group after 48 hr. The LC5 group also returned lower ALH values (p .05) than these seen for the control groups after 48 hr. The fertilizing capacity of LC5 samples stored at 15 degrees C for 2 hr (LC5-15 degrees C-2h) and at 5 degrees C for 24 hr (LC5-5 degrees C-24h) was tested in three ewe groups via cervical fixed-time artificial insemination. In two groups, the fertilizing capacity of the LC5-5 degrees C-24h was reduced (p < .001). In the remaining group, however, no significant difference was seen between the LC5-15 degrees C-2h and LC5-5 degrees C-24h sperm in this respect (pregnancy rates 52.4% versus 42.8%; p > .05). Overall, the present results suggest that supplementing skimmed milk-egg yolk-based extenders with LC has a positive effect on chilled sperm variables and fertilizing capacity.