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Iron derivatives from casein hydrolysates as a potential source in the treatment of iron deficiency.

The properties of an Fe(3+)-peptide complex containing 5.6% Fe, obtained by the reaction of ferric chloride with an enzymatic hydrolysate of casein, are described. The major site of iron binding corresponds primarily to the carboxylate groups and to a lesser extent to the peptide bonds. The Fe(3+)-peptide complex is insoluble at acid pH and completely soluble at neutral to alkaline pH. When soluble, the Fe(3+) is tightly bound to the complex peptide mixture but can be displaced and complexed by a low molecular weight ligand such as cysteine. Its efficacy in relation to iron sulfate was compared in rats. Both iron sources were administrated in Milli-Q water by gastric gavage to male Wistar rats (180-200 g) after an 18 h fast with water ad libitum. Fe(3+) from the Fe(3+)-peptide complex was transferred to the blood in a dose-dependent manner (1-8 mg of Fe/kg), and the serum iron levels were significantly higher (p < 0.001) than in a similar group of rats treated with iron sulfate. In the comparative kinetics experiments, the rats received 4 mg of Fe/kg. Both iron sources presented maximum absorption, as indicated by the elevation of serum iron levels, 30 min after administration, and the AUC(0)(-->2h) of the Fe(3+)-peptide complex was significantly higher (p < 0.05) than that observed with iron sulfate. The simultaneous administration of free peptides (0-192 mg) with the Fe(3+)-peptide complex or iron sulfate did not modify the extent of absorption of iron from both sources, suggesting that the absorption is due to the complex formed and probably not to exchange reactions in the gastrointestinal tract. In the hemoglobin repletion experiments carried out on newly weaned rats with anemia induced by a low-iron diet, supplementation of the diet with the the Fe(3+)-peptide complex was as efficient as supplementation with iron sulfate in the conversion from diet to hemoglobin iron. These results, taken together, suggest that the Fe(3+)-peptide complex is a potential compound for use as an iron source in biological situations.

Anemia, Iron-Deficiency↗

Hemoglobin stimulates the release of proinflammatory cytokines from leukocytes in whole blood.

Isolated mononuclear leukocytes, when incubated with purified hemoglobin Ao (HbAo), release the proinflammatory cytokines interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha). In this study we examined whether leukocytes in whole blood, when incubated with HbAo, release IL-8, TNF-alpha, and IL-6. Leukocytes in whole blood incubated with HbAo for 4 hours at 37 degrees C, 5% CO2, and 95% humidity released 187, 1313, and 50 pg/mL of IL-6, IL-8, and TNF-alpha, respectively, as compared with 6, 192, and 2 pg/mL released by leukocytes in blood incubated with human serum albumin (HSA). Furthermore, plasma from blood incubated with HbAo exhibited chemotactic activity and stimulated human umbilical vein endothelial cells to become adherent to neutrophils. These activities were 3.3 and 2.6 times those measured in plasma from blood incubated with HSA. Hydrocortisone (0.05 micromol/L to 50 micromol/L) inhibited cytokine release in a dose-dependent manner with ED50 values of 0.23 micromol/L, 0.19 micromol/L, and 0.10 micromol/L for IL-6, IL-8, and TNF-alpha, respectively. The release of proinflammatory cytokines in whole blood after exposure to hemoglobin solutions is consistent with the possibility that an inflammatory reaction could develop on infusion of hemoglobin, whereas inhibition of cytokine release by hydrocortisone suggests that the inclusion of anti-inflammatory compounds in hemoglobin solutions may prevent undesirable effects caused by inflammation after infusion.

Blood Substitutes↗

Determinants of benzo(a)pyrenediol epoxide adducts to haemoglobin in workers exposed to polycyclic aromatic hydrocarbons.

OBJECTIVE: The aim was to assess the determinants of benzo(a)pyrenediol epoxide adducts to haemoglobin (BaPDE-Hb) in workers exposed to polycyclic aromatic hydrocarbons (PAHs). METHODS: This was a study of the correlations between the concentrations of PAHs in air, 1-hydroxyprene in urine, and BaPDE-Hb adduct concentration in the blood in 206 men working in steel foundries and one graphite electrode producing plant, taking into consideration confounding factors such as smoking and dietary habits. RESULTS: BaPDE-Hb adduct concentration was correlated (r = 0.26; p = 0.0002) with the airborne PAH concentration and was influenced by tobacco consumption but not by dietary habits. Benzo(a)-pyrene concentration in air, 1-hydroxypyrene concentration in urine, and duration of exposure to PAHs were not associated with the adduct concentration. CONCLUSION: Although environmental exposure to PAHs was statistically associated with BaPDE-Hb adduct concentration, differences between individual subjects in the metabolism of benzo(a)pyrene probably play an important part in determining the amount of BaPDE-Hb adducts formed.

Air↗

Acute myocardial infarction in hemoglobin SC disease.

Although there are reports of myocardial infarction (MI) in patients with sickle cell diseases, an antemortem diagnosis of acute MI in a patient with compound heterozygous hemoglobin SC disease has not been reported. Herein, we present a patient with hemoglobin SC who suffered an acute MI. She had typical chest pain for myocardial ischemia, associated with ST elevations on the electrocardiogram (EKG) and elevations of cardiac injury markers diagnostic of infarction. The patient was treated with conventional therapies for acute coronary syndrome and also emergent red blood cell exchange. Interestingly, coronary angiography was completely normal in this patient. Potential mechanisms and management for acute MI in patients with sickle cell disease are discussed.

Adult↗

Interactions between hemoglobin and organic phosphates investigated with 31P nuclear magnetic resonance spectroscopy and ultrafiltration.

1. The chemical shifts (delta) of the phosphates of 2,3-diphosphoglycerate and adenosine triphosphate (ATP) were determined by phosphorus nuclear magnetic resonance (31P NMR) spectroscopy and were found to be displaced downfield following the addition of hemoglobin (3 mM) to a solution of either diphosphoglycerate (5 mM) or ATP (1 mM). 2. The binding of these compounds to hemoglobin was also determined by membrane ultrafiltration. A direct relationship was observed between the change in chemical shift ((delta delta) of the 2-P and 3-P of diphosphoglycerate and the percent diphosphoglycerate bound, when the latter was varied by altering pH, oxygenation state, or total diphosphoglycerate concentration. 3. In comparable studies with ATP binding, a linear relationship between the delta delta values of the gamma-, beta-, and alpha-P of ATP and the percent of ATP bound was not observed when the data from all of the experiments were plotted. NMR signals were not detectible in deoxyhemoglobin solutions containing 1 mM ATP but were seen in solutions containing 3.8 mM ATP. 4. The results indicate that 31P NMR spectroscopy is a promising tool for investigating organic phosphate interactions with hemoglobin.

Adenosine Triphosphate↗

[Protective action of 5-hydroxy-6-methyluracil and ferrocenes in nitrite-ion poisoning].

It is shown that ferrocenes and 5-oxy-6-methyluracil efficiently inhibit methemoglobin-formation in experiments in vitro and in vivo. Preliminary administration of ferrocene or 5-oxy-6-methyluracil to laboratory animals essentially increases life time in case of sodium nitrite intoxication. Ferrocene protection from sodium nitrite intoxication normalizes the level of methemoglobin and lipoperoxidation in erythrocytes as well as activity of superoxide dismutase of erythrocytes. Activity of catalase, glutathione reductase and dehydrogenases of the pentosephosphate way were not normalized during ferrocene protection from sodium nitrite intoxication. In case of ferrocene protection the catalase activity during sodium nitrite intoxication remains lowered, while activity of glutathione reductase and dehydrogenases of the pentosephosphate pathway grows sharply.

Animals↗

Induction of erythroid differentiation in murine virus infected eythroleukemia cells by highly polar compounds.

Murine-virus-infected erythroleukemia cells cultured in a medium with dimethylsulfoxide or N,N-dimethylformamide are induced to differentiate to erythroid cells. A number of highly polar compounds have a similar effect in inducing erythroid differentiation of the virus-infected cells, as assayed by the appearance of hemoglobin. These compounds are 1-methyl-1-2-piperidone, N,N-dimethylacetamide, N-methylpyrrolidinone, N-methylacetamide, 2-pyrrolidinone, propionamide, pyridine-N-oxide, piperidone, N-methylformamide, acetamide, and triethylene glycol. It has been previously reported that dimethylsulfoxide must be present during DNA synthesis and, possibly, shortly therafter, to induce differentiation. These findings are consistent with the hypothesis that dimethylsulfoxide and related polar compounds act by changing the conformation of DNA or a DNA-protein complex, causing an alteration in transcription that leads to the expression of the program of erythroid differentiation.

Acetamides↗

Bacterial endotoxin (lipopolysaccharide) stimulates the rate of iron oxidation.

Bacterial endotoxin (lipopolysaccharide) has affinity for a number of cations, including iron. Previous investigations have demonstrated that lipopolysaccharide can affect the oxidation rate of iron; heme-bound ferrous iron in hemoglobin is oxidized to ferric iron when hemoglobin binds lipopolysaccharide. In the present study, we directly examined the interaction between lipopolysaccharide and iron. Lipopolysaccharide caused a concentration-dependent increase in the rate of iron oxidation, with up to a 23-fold increase in oxidation in the presence of 200 microg/ml Escherichia coli lipopolysaccharide. This effect was seen both with several carbohydrate-rich smooth lipopolysaccharides and also with carbohydrate-poor rough lipopolysaccharide. Extensively deacylated rough lipopolysaccharide had no effect, suggesting a role of the fatty acid components of lipopolysaccharide in this process. Purified lipid A produced inconsistent results: some preparations stimulated iron oxidation and others did not. A series of sugars, starches and a preparation of purified O-chain polysaccharide (the carbohydrate portion of the lipopolysaccharide macro-molecule) had no effect on the rate of iron oxidation, whereas phospholipid-enriched brain tissue extracts (similar to the lipid A component of lipopolysaccharide) stimulated oxidation. We conclude that the lipid moiety of bacterial lipopolysaccharide is responsible for the stimulation of iron oxidation. This process may contribute to the ability of lipopolysaccharide to cause oxidation of heme-bound iron in hemoglobin.

Carbohydrates↗

Isolation of intermediate valence hybrids between ferrous and methemoglobin at subzero temperatures.

Quenching a hemoglobin solution partially saturated with carbon monoxide into a hydro-organic solvent containing ferricyanide will produce under suitable conditions a population of partially oxidized and CO-bound hemoglobin molecules. Since each Fe3+ heme carries one extra charge, it should be possible, in theory, to resolve the spectrum of intermediate compounds between hemoglobin and carbon monoxide, which was originally present in solution. In this study we report: 1) the development of a simple and rapid method to quench aqueous hemoglobin solutions into a hydro-organic solvent at subzero temperatures; 2) the determination of suitable experimental conditions to isolate valence hybrids between carbonmonoxy- and methemoglobin by isoelectric focusing at temperatures as low as -25 degrees C; and 3) the identification and isolation of all valence hybrids of different charge between carbonmonoxy- and methemoglobin.

Freezing↗

Synthesis and antimalarial effects of phenothiazine inhibitors of a Plasmodium falciparum cysteine protease.

Acridinediones have previously been shown to have potent antimalarial activity. A series of sulfur isosteres of acridinediones have been synthesized and evaluated for their inhibition of the Plasmodium falciparum cysteine protease falcipain and for their antimalarial activity. A number of these phenothiazines inhibited falcipain and demonstrated activity against cultured P. falciparum parasites at low micromolar concentrations. We propose that the compounds exerted their antimalarial effects by two mechanisms, one of which involves the inhibition of falcipain and a consequent block in parasite degradation of hemoglobin. These compounds and related phenothiazines are worthy of further study as potential antimalarial agents.

Animals↗