When polymorphonuclear leukocytes (PMNL) and soluble or particulate matter interact, the cells produce chemiluminescence. Luminol-dependent light emission from PMNL is linked to the myeloperoxidase (MPO)-H202 system. Light emission from a cell-free MPO-H202 system was found to be totally inhibited by human serum albumin (HSA), and since HSA is a large molecular protein that does not readily gain access to intracellular sites of PMNL, it could be used to determine the importance of extra-and intracellular events in PMNL chemiluminescence. In studies with cells from an MPO-deficient patient, we found that HSA inhibited more than 90% of extracellularly produced chemiluminescence. The chemotactic peptide formylmethionylleucyl-phenylalanine induced a two-peak chemiluminescence response in normal PMNL, and addition of HSA reduced the first peak, whereas the second peak was unaffected. This result indicated that the first peak was a result of extracellular reactions and the second peak was a result of intracellular reactions of the MPO-H202 system. Most of the phorbol myristate acetate-induced response in normal PMNL was due to intracellular events. Furthermore, chemiluminescence of intracellular origin seems to be limited not by generation of oxidative metabolites but by diffusion of luminol into the cells.
Background The most common complications of prosthetic hip joints are aseptic mechanical failure and infection. Delayed low-grade infections are seen most often, and they are also most difficult to distinguish from aseptic mechanical failures.Methods We conducted a prospective study to compare inflammatory markers in patients diagnosed with aseptic or septic prosthetic loosening. The diagnostic criteria were based on the decisions of experienced orthopedic surgeons and microbiological analysis of periprosthetic tissue samples taken perioperatively.Results Coagulase-negative staphylococci were the commonest pathogens in the infected patients. Pre-or perioperative elevation of C-reactive protein and erythrocyte sedimentation rate were significantly greater in the infection group, as were white blood cell count and levels of cytokines in synovial fluid. The patterns of infiltration of inflammatory cells in periprosthetic tissue were also significantly different between the groups.Interpretation A combination of clinical judgment and multiple tissue samples constitutes a good platform for distinguishing between septic and aseptic loosening of prostheses. Moreover, the combined use of several laboratory and histopathological markers of inflammation, especially infiltration of polymorphonuclear cells, further helps the diagnosis.
This study demonstrates the kinetic phases of viremia during PHI and indicates two new contributions to the natural history of HIV-1 infection: PHI peak levels correlate with steady-state levels and HIV-1 RNA declines biphasically; an initial rapid decay is usually followed by a slow decay, which is similar to the initial changes seen with antiviral treatment.
Exudate human polymorphonuclear neutrophils were isolated and investigated regarding oxidative responsiveness and priming ability. The exudate neutrophils were found to produce an increased amount of O2- and H2O2 when stimulated with formylmethionyl-leucyl-phenylalanine (fMLP), i.e. these cells were metabolically primed. Cytochalasin B (cyt B) pretreatment affected the production of O2- by exudate cells, although to a lesser extent than the production by peripheral blood cells, in which a substantial increase was induced. Addition of N-ethylmaleimide (NEM) to activated exudate and peripheral blood cells revealed no difference in oxidase inactivation rate. To induce further priming, the cells were incubated in vitro with a synthetic diacylglycerol (sn-1,2-didecanoylglycerol; diC10), or the Ca2+ ionophore ionomycin. Results of this procedure showed significant differences between exudate and peripheral blood neutrophils: the peripheral cells expressed a primed response, which was measured as increased fMLP-induced O2- production following incubation with both these substance; whereas the metabolic activity of exudated cells was not affected by diC10, but was significantly primed by ionomycin (P less than 0.01). The exact route for diacylglycerol priming is unknown. However, our results with human neutrophils primed during exudation indicate an exhausted diC10-priming pathway, with a retained sensitivity for priming [Ca2+]i rises.
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