Inhibition of the heme-induced hemolysis of red blood cells by thechlorite-based drug WF10.

Excessive release of hemoglobin from red blood cells markedly disturbs the health status of patients due to cytotoxic effects of free hemoglobin and heme. The latter component is able to initiate novel hemolytic events in unperturbed red blood cells. We modeled this process by incubation of ferric protoporphyrin IX with freshly isolated red blood cells from healthy volunteers. The heme-induced hemolysis was inhibited in a concentration-dependent manner by the chlorite-based drug WF10, whereby the hemolysis degree was totally abolished at a molar ratio of 1:2 between chlorite and heme. Upon incubation of heme with WF10, the ultraviolet-visible spectrum changed, whereas the release of iron from heme and the appearance of fluorescent breakdown products of the porphyrin ring were negligible at this ratio, but increased with increasing excess of chlorite over heme. Thus, inhibition of hemolysis by WF10 takes already place at those chlorite concentrations, where no degradation of the porphyrin ring occurs. As WF10 is applied in form of an intravenous infusion to patients with severe inflammatory states, these data support the hypothesis that the beneficial WF10 effects are closely associated with inactivation of free heme.

Randomized, double-blind, placebo-controlled trial of the immune modulator WF10 in patients with advanced AIDS.

A randomized, double-blind trial compared treatment with the immune modulator WF10 (ten patients) and placebo (nine patients) administered in cycles over 3 months among individuals with advanced AIDS. There were no notable clinical adverse events; changes in hematologic and chemistry values were comparable in the two groups. In both groups, median HIV-RNA PCR values remained stable. Immunologic variables showed a consistent tendency to increase in the WF10 group and to decrease in the control group, with significant differences between groups for median WBC, lymphocyte, CD19, and CD35 values. Ten infections occurred in the control group, four of which were Pneumocystis carinii pneumonia (PCP), and three in the WF10 group none of which was PCP. Five patients in the control group were hospitalized during the trial for a total of 53 days; no patients in the WF10 group were hospitalized. Over a subsequent 9-months follow-up, six patients from the control group and one from the WF10 group died. These results indicate that WF10 administration appears safe, may enhance immunologic function, and unlike other macrophage-activating cytokines does not increase HIV expression in this patient population. Further studies of WF10 in larger patient populations are warranted.

Inactivation of HIV infectivity by the chlorite-oxygen reaction product tetrachlorodecaoxygen.

OBJECTIVE: Since the chlorite-oxygen reaction product tetrachlorodecaoxygen (TCDO) anion complex promotes efficaciously tissue repair and has antibacterial activity, our aim was to determine the effects of TCDO on the replication of HIV and on the infectivity of free HIV particles. DESIGN: The effects of TCDO on cellular HIV replication machinery and the consequences of TCDO for infectivity of HIV virions were evaluated. METHODS: Virus yields in supernatants of TCDO-supplemented cultures of HIV-infected cells or virus infectivity in TCDO-treated virus stocks were quantified by titration assays and then calculating the 50% tissue culture infectious dose. RESULTS: First, TCDO did not affect the replication of HIV in persistently infected lymphocytic and monocytic cell lines or in peripheral blood mononuclear cells. Second, supplementation of HIV stocks with TCDO markedly decreased the infectivity of HIV particles in a concentration dependent manner. Third, the binding of gp120 envelope glycoprotein of HIV-1 to cells is blocked by pre-incubation with TCDO. Fourth, the inhibition of HIV replication by the reverse transcriptase inhibitor 3'-azido-3'-deoxythymidine (zidovudine) in de novo infected cell cultures was not affected by the simultaneous addition of TCDO. However, the delayed virus spread of HIV in cultures in the presence of suboptimal concentrations of zidovudine could significantly be blocked by the simultaneous addition of TCDO. Fifth, TCDO failed to induce the chromosomally integrated HIV-1 provirus in the T-lymphoma cell line ACH2. CONCLUSIONS: TCDO appears to inactivate HIV particles directly, but has no influence on the intracellular replicative machinery of HIV. Our results suggest that a clinical evaluation of the TCDO complex as chemotherapy for HIV infection and full-blown AIDS should be considered, particularly in patients concomitantly receiving zidovudine.

Increased resistance towards two systemic experimental infections by tetrachlorodecaoxygen anion complex. Possible implications of cellular and humoral immunity.

The tetrachlorodecaoxygen anion complex (TCDO, Oxoferin) given intravenously 1 h after intravenous infection of mice with Candida albicans increased the host's resistance gradually in doses up to 3.1 mumol/kg body weight (b.w.). Above this level, the stimulatory effect decreased, turning to an inhibition at a dose of 18.6 mumol/kg b.w. TCDO. Repeated applications of the optimum single dose had no cumulative effect. In experimental infections with the strictly anaerobic Peptostreptococcus intermedius, TCDO ameliorated the course of the infection not only at a dose of 3.1 mumol/kg b.w. but also at a higher dose of 12.4 mumol/kg b.w. TCDO, which in the Candida sepsis model fell into the range of defence inhibition. A single dose of 3.1 mumol/kg b.w. TCDO also revealed to be the optimum dose to increase the humoral immune response evaluated by the number of immunoglobulin (Ig)M and IgG forming spleen cells after sensitization with sheep red blood cells (SRBC). The same results were obtained, regardless of whether TCDO had been given at the time of immunization or thereafter. The higher single dose of 18.6 mumol/kg b.w. TCDO did not show this effect but was not inhibitory either. Cellular immune reactions evaluated by the footpad swelling test and SRBC as antigen were found considerably enhanced whether TCDO was given once or repeatedly at a dose of 3.1 mumol/kg b.w., whereas single doses of 18.6 mumol/kg or repeated doses of 9.3 mumol/kg induced a certain inhibition.(ABSTRACT TRUNCATED AT 250 WORDS)

Time kinetics of hemoglobin and myoglobin activation by tetrachlorodecaoxide (TCDO).

In the presence of peroxidase, myoglobin or hemoglobin, Tetrachlorodecaoxide (TCDO) forms an active oxygen species which is similar to the product of the polymorphonuclear leucocyte (PMNL) myeloperoxidase reaction and the 'Klebanoff Model' of phagocytosis, but it is also produced under anaerobic conditions. Randomly destructive species such as the free OH radical or singlet oxygen are not formed. The kinetics of the heme-dependent activation vary according to the heme type present. In comparison to myoglobin, blood shows a 2 h delay in the appearance of maximal activity. On the basis of known biochemical and clinical-physiological data, a hypothesis can be proposed to explain the reoxygenation observed in hypoxic tissue, induced by TCDO via this activated heme species. Under normal physiological conditions, vasodilation occurs via catalysis by xanthine oxidase or PMNL-dependent activation of fatty acids.

Biochemical oxygen activation as the basis for the physiological action of tetrachlorodecaoxide (TCDO).

Oxidation of methionine, 4-(methylthio)-2-oxobutyric acid (KMB), or 1-aminocyclopropane carbonic acid (ACC) are indicator reactions for activated oxygen species such as singlet oxygen (1O2), OH.-radical like oxidants, superoxide anion (O2.-), hydrogen peroxide (H2O2) or activated hemo-iron complexes like peroxidase- or catalase-"compound I". Methionine is oxidized by OH. as well as by 1O2 forming ethylene, but not by tetrachloro-decaoxygen complex (TCDO) in the absence or presence of catalytic hemoproteins such as peroxidase, hemoglobin or myoglobin. Both KMB and ACC are oxidized by TCDO under the catalysis of the above hemo-proteins where neither catalase nor superoxide dismutase are inhibitors. TCDO hemo-protein complex is an oxidant with similar properties as peroxidase-compound I and can clearly be differentiated from O2.-, H2O2, OH. and 1O2.

New aspects on the pathophysiology of wound infection and wound healing--the problem of lowered oxygen pressure in the tissue.

There is a correlation between tissue oxygen tension, incidence of infection and disturbance of wound healing. Scientific investigations in recent years have documented that infection and tissue repair are processes consuming oxygen. Animal experiments have shown that oxygenation of the tissue is essential for the elimination of pathogens, the stimulation of phagocytosis as well as for the degradation of dead tissue structures and the synthesis of new tissue structures. Hypoxia and hypovolemia are of crucial significance for the impairment of organ functions. An effective therapy for disorders of oxygenation has not been feasible so far, except for clinical anesthesiological efforts to increase the oxygen saturation of hemoglobin and hyperbaric oxygen therapy. However, it has recently been possible to synthesize an oxygen carrier which supports respiratory oxygen physiologically. With tetrachlorodecaoxide (TCDO), a causal therapeutic concept for the topical treatment of infected hypoxic wounds with chronic disturbance of wound healing has been introduced for the first time.

In vitro investigations on the antibacterial action and the influence on the phagocytic chemiluminescence of tetrachlorodecaoxide--a new, non-metallic oxygen complex.

The first non-metallic oxygen carrier, tetrachlorodecaoxide (TCDO), showed in vitro antibacterial activity among aerobic and anaerobic bacteria. The lethal dose for Escherichia coli, for example, was 150 micrograms/ml whereas 15 micrograms/ml reduced the bacterial amount after a latent period of two hours, but regrowth started after four hours. The bactericidal effect of TCDO, however, was dose-dependent and species-specific. This suggests that some aerobic bacterial species might not be able to produce sufficient amounts of protecting enzymes like catalase or superoxide dismutase. The computer controlled measurement of chemiluminescence was used as a model for the phagocytic activity. With isolated human granulocytes and opsonized zymosan as antigen no increase in peak counts per minute was observed compared with controls without TCDO. However, with human whole blood, positive effects were seen using TCDO together with zymosan as well as specific and non-specific opsonized Klebsiella pneumoniae K 17. It seems that whole blood possesses additional, but as yet unknown biocatalysers to split TCDO into oxygen and chloride.

[Stimulation of wound healing by tetrachlordecaoxide. Results of a randomized double-blind study]. / Stimulation der Wundheilung durch Tetrachlordecaoxid (TCDO). Ergebnisse einer randomisierten Doppelblindstudie.

In 38 patients with chronic therapeutically resistant wounds, which, in 25 cases, had been existing for more than one year, Tetrachlorodecaoxide ( TCDO ) in a water solution containing glycerin was analyzed for its capacity to induce wound healing and compared in this respect to the standard in moist wound treatment, physiological sodium chloride. The results of the clinical trial demonstrate that the TCDO solution is significantly superior to physiological saline in local wound treatment regarding the degree of wound smear reduction, the formation of wound granulation tissue, the stimulation of epithelisation on the wound borders and the shrinking of the wound surface. The differences in therapeutic efficiency are so large that, in spite of the relatively small patient samples (21 + 17) it was possible to verify the superiority of a method for wound treatment in a randomized double blind clinical trial.

In vitro investigations on the antibacterial action and the influence on the phagocytic chemiluminescence of tetrachlorodecaoxide--a new, non-metallic oxygen complex.

The first non-metallic oxygen carrier, tetrachlorodecaoxide (TCDO), showed in vitro antibacterial activity among aerobic and anaerobic bacteria. The lethal dose for Escherichia coli, for example, was 150 micrograms/ml whereas 15 micrograms/ml reduced the bacterial amount after a latent period of two hours, but regrowth started after four hours. The bactericidal effect of TCDO, however, was dose-dependent and species-specific. This suggests that some aerobic bacterial species might not be able to produce sufficient amounts of protecting enzymes like catalase or superoxide dismutase. The computer controlled measurement of chemiluminescence was used as a model for the phagocytic activity. With isolated human granulocytes and opsonized zymosan as antigen no increase in peak counts per minute was observed compared with controls without TCDO. However, with human whole blood, positive effects were seen using TCDO together with zymosan as well as specific and non-specific opsonized Klebsiella pneumoniae K 17. It seems that whole blood possesses additional, but as yet unknown biocatalysers to split TCDO into oxygen and chloride.

[Experimental model of protracted traumatic-hemorrhagic shock. I. Organ blood flow]. / Experimentelles Modell des protrahierten traumatisch-hämorrhagischen Schocks: I. Organdurchblutung.

Experimental trauma and hemorrhage acutely reduce capillary blood flow in all organs except the heart. After reinfusion of the withdrawn blood a long-lasting increase of pulmonary bronchial blood flow and hepatic and renal blood flow is observed parallel to a rise in cardiac output. However, blood flow in the intestines, pancreas, and spleen remains significantly reduced. Particularly, renal and pulmonary bronchial blood flow is significantly higher in animals surviving for 72 hr than in animals succumbing after an average of 32 hr.

Experimental gastric sympathectomy: an effective prophylaxis of gastric stress lesions.

Fifteen mini-pigs were bled to a mean aortic blood pressure of 40 mm Hg and maintained at that level for three hours. The control group consisted of eight animals with shock for three hours, while the test group was comprised of seven animals with a similar shock period but which had undergone splanchnicectomy 14 days earlier. In all animals a stimulated gastric secretion test was performed three days before and eight days after splanchnicectomy. All animals in the control group showed severe gastric mucosal lesions after shock. Conversely, the piglets with splanchnicectomy developed no changes (five animals) or only minor changes (two animals). The efficacy of splanchnicectomy was confirmed by a stimulated gastric secretion test in which basal acid output did not change after operation, but peak acid output increased significantly. This study suggests that gastric splanchnicectomy prevents gastric ulceration following experimental shock.