Elaine M. Berger

Intact human erythrocytes prevent hydrogen peroxide-mediated damage to isolated perfused rat lungs and cultured bovine pulmonary artery endothelial cells.

Addition of untreated or glutaraldehyde-fixed human erythrocytes decreased hydrogen peroxide (H2O2)-mediated acute edematous injury in isolated rat lungs, H2O2-induced damage to cultured bovine pulmonary artery endothelial cells, and H2O2-dependent oxidation of reduced cytochrome C in vitro. The results suggest that intact erythrocytes can scavenge H2O2, and as a result, protect the lung and possibly other tissues from damage.

Iron depletion or chelation reduces ischemia/reperfusion-induced edema in gerbil brains☆

Since hydrogen peroxide (H2O2) can react with ferrous iron (FE++) to form the more toxic hydroxyl radical (OH) in vitro, and since H2O2 is generated brain xanthine oxidase (XO) during ischemia/reperfusion (I/R), we hypothesized that gerbils depleted of iron by dietary restriction or treated with iron chelators would be less susceptible to I/R injury. We found that gerbils fed a low iron diet for 8 weeks had decreased brain and serum iron levels, less neurologic deficits, and decreased brain edema after temporary unilateral carotid ligation (ischemia) and then reperfusion than gerbils fed a control standard iron diet. In addition, brains from gerbils treated with iron-free deferoxamine (an iron chelator), but not iron-loaded deferoxamine, had decreased (P less than .05) brain edema following ischemia and reperfusion. The results indicate that iron may contribute to cerebral ischemia/reperfusion damage.

Hydrogen peroxide causes dimethylthiourea consumption while hydroxyl radical causes dimethyl sulfoxide consumption in vitro

Addition of increasing concentrations of hydrogen peroxide (H2O2) caused progressive decreases in dimethylthiourea (DMTU) concentrations which were inhibitable by simultaneous addition of catalase, but not the superoxide anion (O2-.) scavenger, superoxide dismutase (SOD), or hydroxyl radical (.OH) scavengers, such as mannitol, sodium benzoate or dimethyl sulfoxide (DMSO). In parallel, addition of increasing concentrations of H2O2 with FE++/EDTA (but not H2O2 alone) caused decreases in DMSO concentrations which were inhibitable by simultaneous addition of .OH scavengers but not SOD or catalase. Addition of DMTU, but not DMSO, also decreased H2O2 concentrations in vitro. The results indicate the relative scavenging specificities of DMTU and DMSO for H2O2 and .OH, respectively. The findings also suggest that measurement of DMTU or DMSO consumption could help assess the contribution of O2 metabolites in biological systems.

Hepes may stimulate cultured endothelial cells to make growth-retarding oxygen metabolites

SummaryZwitterion buffers are often used to modulate the pH of cell culture medium but their effect on cultured cells is controversial. We found that addition of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) caused superoxide dismutase (SOD) inhibitable increases in nitroblue tetrazolium dye reduction and SOD and catalase inhibitable decreases in the growth of cultured bovine pulmonary artery endothelial cells. The findings suggest that HEPES stimulates endothelial cells to make toxic oxygen metabolites that contribute to decreased cell growth.

Increased serum catalase activity in rats subjected to thermal skin injury

We found that: rats subjected to thermal skin injury (burn) had increased serum hydrogen peroxide (H2O2) scavenging activity, serum catalase activity, erythrocyte (RBC) fragility, and edematous lung injury (lung leak) when compared to sham-treated rats. Serum H2O2 scavenging activity was inhibited by addition of sodium azide, a catalase inhibitor. Treatment of rats with the oxygen radical scavenger, dimethylthiourea (DMTU), decreased RBC fragility and lung leak but did not alter increased H2O2 scavenging or catalase activity of serum from rats subjected to skin burn. We conclude that increased serum catalase activity is a consequence of thermal skin injury and that increased serum catalase activity may be a mechanism that modulates H2O2-dependcnt processes following skin burn.

Human phagocytic cells as oxygen metabolite scavengers.

Human neutrophils or monocytes decreased hydrogen peroxide (H2O2) concentrationsin vitro. Neutrophils or monocytes decreased H2O2 concentrations as well as human erythrocytes. Treatment with aminotriazole or azide decreased both phagocyte and erythrocyte catalase activity and the ability of each cell to decrease H2O2 concentrationsin vitro. Prestimulation of phagocytic cells with phorbol myristate acetate (PMA) or opsonized zymosan decreased neither their catalase activity nor their ability to decrease H2O2 concentrations. The results suggest that unstimulated or stimulated phagocytic cells can scavenge H2O2 and may potentially decrease H2O2-mediated tissue injury. The H2O2 scavenging potential of phagocytic cells is due at least partially to their catalase activity.

Increased incidence of menstruation-associated bactericidal defects in neutrophils from women who have recovered from toxic shock syndrome

There is a growing suspicion that a host abnormality may contribute to the pathogenesis of toxic shock syndrome (TSS). We found that females (5 of 5) who had recovered from TSS had transient, menstruation-associated decreases (≥9%) in the ability of their neutrophils to killStaphylococcus aureus, 502A in vitro more often (P=0.040 by Fishers exact test) than non-TSS-affected control subjects (5 of 12). In addition, the average decrease in bactericidal activity in neutrophils obtained during menstruation from recovered TSS patients was 30 ± 9% compared to 7 ± 7% for neutrophils from non-TSS-affected control subjects. The results are consistent with the possibility that transient menstruation-associated decreases in neutrophil bactericidal function may indicate susceptibility and/or contribute to the development of TSS.

Effect of dimethyl sulfoxide on the bactericidal function of polymorphonuclear leukocytes.

Abstract : Dimethyl sulfoxide (DMSO) is now being widely used to treat various rheumatic, musculoskeletal, and inflammatory conditions, but the therapeutic and detrimental consequences of treating individuals with DMSO remain unclear. We have examined the possibility that DMSO might inhibit the bactericidal function of polymorphonuclear leukocytes (PMN) by scavenging hydroxyl radical (.OH), one of the key bactericidal components of PMN. Our results suggest that DMSO can decrease the ability of PMN to kill Staphylococcus aureus in vitro and that this inhibition may occur because DMSO scavenges .OH. These observations suggest that DMSO might impair PMN-mediated host defense.

Variable effect of toxic shock toxins from different sources on neutrophil function in vitro

Toxic shock syndrome toxins (TSST) are 23–30 kD proteins that have been isolated from incubation media of strains ofStaphylococcus aureus cultured from patients with toxic shock syndrome (TSS). Injection of TSST into animals produces many of the symptoms that characterize TSS including shock, fever, and multiple organ failure. We found that addition of increasing concentrations of TSST-1-VP1035-16A, but not TSST-PEC, TSST-SEC, staphylococcal enterotoxin A or B, progressively decreased human neutrophil bactericidal activity againstS. aureus, 502A in vitro. TSST-1-VP1035-16A, but not the other toxins, also decreased superoxide anion and hydrogen peroxide concentrations in mixtures containing neutrophils and phorbol myristate acetate (PMA) in vitro. The results indicate that various preparations of TSST have different effects on neutrophil function in vitro and, accordingly, may have different effects in other in vitro and in vivo models of TSS.

Persistent bactericidal defect in neutrophils from a young woman who recovered from toxic shock syndrome

We have previously found transient menstruation-associated abnormalities in the in vitro bactericidal function of neutrophils from females who have recovered from toxic shock syndrome (TSS). We now report the case of a young woman who has also recovered from TSS, but who has a persistent, non-menstruation-associated defect in the ability of her neutrophils to killStaphylococcus aureus in vitro.