Paul H. Gale

Coenzyme Q. XVII. Isolation of coenzyme Q10 from bacterial fermentation

Abstract Crystalline coenzyme Q10 has been newly isolated from a fermentation using Pseudomonas denitrificans. Previously, the presence of Q9 in Pseudomonas fluorescens, and Q10 in Neurospora crassa and in Chromatium was reported by others, but without evidence based on characterization of isolated crystalline material. We found Q9 in five other species of Pseudomonas, Q8 and Q9 in Pseudomonas fluorescens, and evidence for the presence of some form of Q in 32 cultures, representing 20 species out of 107 cultures which were examined. Coenzyme Q10 can now be produced by a suitable fermentation, rather than by isolation from mammalian tissue.

Radical scavenging as the mechanism for stimulation of prostaglandin cyclooxygenase and depression of inflammation by lipoic acid and sodium iodide

Certain radical-trapping reducing agents have been shown to stimulate prostaglandin biosynthesis in vitro (1--6) and to depress phorbol myristate acetate-induced mouse ear edema (16). The increased prostaglandin synthesis resulted from influences on the cyclooxygenase. To ascertain whether these alterations were due to direct interaction with the enzyme or to indirect scavenging of the oxidant released during PGG2 reduction, we report the effects of lipoic acid and sodium iodide. Both of these agents stimulated the enzymatic oxygenation of arachidonic acid, increased the reduction of PGG2 to PGH2, quenched the EPR signal induced by arachidonic acid and depressed mouse ear edema. In addition to discovering two unusual antiinflammatory agents, we have confirmed that materials with entirely different structures can have identical effects on the cyclooxygenase, suggesting indirect stimulation of this enzyme due to trapping of the oxidant.

On the presence and significance of coenzyme Q in microsomes

Mitochondrial and microsomal fractions were prepared from rat liver and particularly from porcine adrenal glands. However, microsomal fractions which are completely free from mitochondrial fragments are very difficult to obtain. Since the succinoxidase enzyme system occurs only in mitochondria, it was possible to determine the percentage of mitochondrial contamination in the microsomal preparations. Such data are necessary in order to estimate the coenzyme Q content of microsomes. Coenzyme Q10 was found in microsomes from porcine adrenal glands, and both Q9 and Q10 were found in the microsomes of rat liver. It appears, therefore, that the coenzyme is implicated in enzyme systems of microsomes as well as those of mitochondria. Further research on adrenal mitochondrial and microsomal enzymic reactions in which coenzyme Q participates may be particularly merited.

Coenzyme Q. XXIV. On the significance of coenzyme Q 10 in human tissues

Abstract Several organs and tissues of three humans have been examined for coenzyme Q content. The liver, heart, spleen, kidney, pancreas, and adrenals contain relatively high concentrations of coenzyme Q 10, indicating that studies of the functional relationship of coenzyme Q to diseases involving any of these organs might be important. The thyroid and brain contain quite low levels of Q. The total body content of coenzyme Q 10 appears to be in the range of 0.5–1.5 g., and the intestinal flora may contribute only negligible amounts of Q 10 to body stores. Coenzyme Q 10 would seem to have an important role in human health and disease, because of (a) its presence in essential organs, (b) its direct link to known vitamin derived coenzymes, (c) its coenzymic functions, and (d) an apparent role in oxidative phosphorylation.

Coenzyme Q. IX. Coenzyme Q9 and Q10 content of dietary components

Abstract The results of coenzyme Q analyses of some common dietary ingredients are reported. Corn oil and wheat germ oil contain considerable quantities of coenzyme Q 9 . Smaller amounts of Q 10 were found in butter, green beans, crude soybeans, soybean oil, and isolated soybean protein product. Other materials tested did not contain measurable amounts of coenzyme Q 10 . These data permit the formulation of Q 10 -low or perhaps Q 10 -free diets.

Coenzyme Q. XIII. Isolation, assay and human urinary levels of coenzyme Q10

Pure coenzyme Q10 has been isolated from normal human male urine; a colorimetric assay for determining Q10-levels in urine has been devised and applied to 160 collections representing 63 males and females. The average excretion is ca. 55 μg./24 hr. for males and 22 μg./24 hr. for females. Thirty-five per cent of the 63 individuals excreted < 10 μg./24 hr. on one or more occasions. Whether or not such low levels of Q10 urinary excretion represent tissue deficiency and possible disease state(s) remains for further research.

Comparative Biochemistry of Lipoxygenase Inhibitors

This chapter describes some studies on the mechanism of 5-lipoxygenase regulation and inhibition and then presents an overview of lipoxygenase inhibitors that are active against intact-cell, broken-cell, and purifed enzymes from a variety of sources. Although a large number of lipoxygenase inhibitors have been reported, many of these are rather nonselective. Some of the lack of selectivity may result from a common mechanism of lipoxygenase inhibition by antioxidants and redox regulation. It is, therefore, not sufficient to define specificity against broken-cell enzymes. A truly diagnostic agent must be specific in cellular systems and in vivo to be of utility in defining the role of lipoxygenases in pathophysiology.

COENZYME Q. LI. NEW DATA ON THE DISTRIBUTION OF COENZYME Q IN NATURE.

The heart of the rhesus monkey was found to contain coenzyme Q 10 . This experimental primate may serve as a model for studies of CoQ 10 as it may relate to human disease. Frog nerve tissue was also found to contain CoQ 10 . Such tissue may be a useful system for researches on CoQ in nerve physiology. Only CoQ 10 was identified in human and rodent tumors grown in mice. The tumor strains were HAd-1, HS-1, and S-180. Cells of Ochromonas malhamensis contain CoQ 10 . It is of interest that both man and this Ochromonas sp. specifically use both CoQ 10 and vitamin B 12 . Contrary to an earlier report that a given Polyporous did not contain a CoQ, it has now been found that Polyporous schweinitzii contains both CoQ 9 and CoQ 10 . The “J strain” of a PPLO ( Mycoplasma gallisepticum ) is of current interest both in respect to its nutrition and its infectious characteristics; this PPLO strain did not contain either a member of either the CoQ or vitamin K groups.

Biochemical and biological activities of 2,3-dihydro-6-[3-(2-hydroxymethyl)phenyl-2-propenyl]-5-benzofuranol (L-651,896), a novel topical anti-inflammatory agent

The biochemical and biological profile of a topical anti-inflammatory agent, 2,3-dihydro-6-[3-(2-hydroxymethyl)phenyl-2-propenyl]-5-benzofuranol (L-651,896 inhibited the 5-lipoxygenase of rat basophilic leukemia cells with an IC50 of 0.1 microM and leukotriene synthesis by human PMN and mouse macrophages with IC50 values of 0.4 and 0.1 microM respectively. L-651,896 also inhibited prostaglandin E2 synthesis by mouse peritoneal macrophages (IC50 = 1.1 microM). This compound inhibited ram seminal vesicle cyclooxygenase activity at considerably higher concentrations, and this effect was directly related to substrate concentration. When applied topically to the mouse ear, L-651,896 lowered elevated levels of leukotrienes associated with arachidonic acid-induced skin inflammation and delayed hypersensitivity induced by oxazolone. However, while L-651,896 inhibited the increased vascular permeability induced by arachidonic acid, it had no effect on the edema associated with the immune-based response to oxazolone in the same tissue. Thus, it is possible that leukotrienes may play a role in some but not all inflammatory responses.

Acetaminophen as a Cosubstrate and Inhibitor of Prostaglandin H Synthase

Recently, several reports (Marnett et al., 1983; Nordenskjold et al., 1984) have implicated prostaglandin H synthase (PHS) in the bioactivation of xenobiotics to potentially toxic metabolites. Benzidine (Zenser et al., 1983), p-aminophenol (Josephy et al., 1983), and phenacetin (Andersson et al., 1982) are among the compounds known to undergo metabolic activation by PHS. Of particular interest to us is the fact that this enzyme can metabolize acetaminophen (APAP) to a reactive species that can bind to proteins or form a glutathione conjugate (Moldeus and Rahimtula, 1980; Boyd and Eling, 1981; Mohandas et al., 1981; Moldeus et al., 1982). In fact, it has been suggested (Boyd and Eling, 1981; Mohandas et al., 1981) that the nephrotoxicity sometimes associated with APAP overdosage may be due in part to its metabolism by PHS which is present in high levels in the renal inner medulla.