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Biomedical subjects

M A Paz

Publications and source records attributed to M A Paz.

At least 19 recordsLinked to original sources
PubMed1996

The catalysis of redox cycling by pyrroloquinoline quinone (PQQ), PQQ derivatives, and isomers and the specificity of inhibitors.

Pyrroloquinoline quinone (PQQ) is a widely distributed redox-active cofactor and essential nutrient. For its detection in protein-free ultrafiltrates or dialysates, a highly sensitive amplification assay was developed on the basis of PQQ's ability to catalyze redox cycling at pH 10 in the presence of excess glycine, oxygen, and nitro blue tetrazolium. Herein, we examine the propensities of PQQ, PQQ triester, and its various isomers, and certain PQQ triester derivatives, to catalyze glycine-fueled redox cycling and show that PQQ is the most capable of catalyzing redox cycling. Furthermore, PQQ has a unique pattern of inhibition induced by a series of PQQ antagonists of different potencies. The data indicate that putative PQQ from a biological sample, separated by HPLC and detected by the glycine-fueled redox-cycling assay, can be further identified as PQQ based on the profile of inhibition it displays with the antagonists such as those employed in this study. The methodology presented here should facilitate the specific detection of PQQ in biological samples.

Animals↗
PubMed1995

Inhibition of redox cycling of methoxatin (PQQ), and of superoxide release by phagocytic white cells.

The iodonium compounds diphenyleneiodonium and diphenyliodonium, and the amine compounds, 4,5-dimethyl phenylene diamine, N,N-dimethyl 1,4-phenylene diamine, 1,2-diamino-4,5-methyleneoxybenzene, and aminomalononitrile inhibit methoxatin's (PQQ's) redox activity in vitro, that is, the methoxatin-coupled oxidation of glycine and reduction of nitroblue tetrazolium to formazan. The compounds mentioned above also inhibit phorbol myristate acetate (PMA) stimulated superoxide release by phagocytic white cells--determined mainly as the superoxide dismutase sensitive reduction of ferricytochrome C. Related compounds, 3,4-diaminopyridine and 4-dimethylamino-benzylamine, did not inhibit redox activity of PQQ in vitro, nor did they inhibit PMA stimulated superoxide production in monocytes or neutrophils. Thus, there is a correlation between an agent's ability to inhibit PQQ redox cycling and its ability to inhibit superoxide release by phagocytes. The findings are a further indication that PQQ is involved in the respiratory burst of phagocytic cells.

Amines↗
PubMed1994

Antisense RNA to the first N-glycosylation gene, ALG7, inhibits protein N-glycosylation and secretion by Xenopus oocytes.

N-Glycosylation has been shown to affect the rate of glycoprotein transport through the secretory pathway. In order to identify the critical components in the N-glycosylation pathway that directly influence protein secretion, we have studied the effects of downregulation of the first gene in the dolichol pathway, ALG7, on the synthesis, glycosylation and secretion of native and heterologous proteins by Xenopus laevis oocytes. Our strategy involved the use of ALG7 antisense RNA (asRNA) to lower the effective abundance of the ALG7 protein in oocytes. The results showed that there was an inverse dose-response relationship between ALG7 asRNA and the amount of glycosylated and secreted proteins. These effects were also observed for heterologously expressed rat parotid amylase. Since ALG7 asRNA did not inhibit overall protein synthesis, we conclude that downregulation of ALG7 expression directly lowered protein export.

Amylases↗
PubMed1994

Methoxatin (PQQ) in guinea-pig neutrophils.

PQQ, also called methoxatin, has been isolated from guinea-pig neutrophils. The organic cations diphenyleneiodonium (DPI) and diphenyliodonium (BPI) and the aromatic o-diamine 4,5-dimethylphenylenediamine (DIMPDA) sequester synthetic PQQ and inhibit its redox-cycling activity in a model system. Standards were made of adducts of tritiated PQQ with unlabeled DIMPDA and of unlabeled PQQ with tritiated DPI or DIMPDA. PQQ adducts were isolated from guinea-pig neutrophils with each of the tritiated inhibitors. They were separated and defined by high-performance liquid chromatography (HPLC). Tiron, a disodium benzene disulphonic acid, broke the DPI-PQQ adduct isolated from neutrophils and released free PQQ. Both DPI and DIMPDA, as well as BPI, blocked O2.- release by stimulated neutrophils. The blockade exerted by these inhibitors was released by the addition of PQQ to the cell suspensions. The data demonstrate the presence of PQQ in guinea-pig neutrophils and suggest that it has a possible role, direct or indirect, in the O2.(-)-producing respiratory burst.

Animals↗
PubMed1994

Aspects of the release of superoxide by leukocytes, and a means by which this is switched off.

Although great progress has been made in understanding the respiratory burst of leukocytes that produce superoxide (O2-), it is possible that a component or components, might have been overlooked. Furthermore, O2- production and its sequels, though cardinal in bactericidal action, might ultimately be damaging to the host's own cells. It is important, therefore, that a biologic mechanism exist to turn off O2- production by stimulated leukocytes. This article offers evidence that methoxatin (PQQ), a redox-cycling orthoquinone, might be involved in O2- production by leukocytes. This is based on the fact that inhibitors of O2- production, such as diphenylene iodonium (DPI) and 4,5-dimethylphenylene diamine (DIMPDA), were shown to sequester PQQ in leukocytes, i.e., to form adducts with that substance. Addition of PQQ to cells blocked with the inhibitors partially restored O2- release. With respect to turning off cellular O2- release, a factor was observed to be released to the medium by old macrophages (14 days old, but not by those less than 7 days old). Such conditioned medium, when added to stimulated neutrophils or macrophages, blocked O2- release. This factor was sensitive to proteases, exhibited molecular sizes of 3 and 11 kDa, and its action was independent of the nature of the stimulus applied to the leukocytes. It was partially purified by column (sizing) chromatography and HPLC. It seems to be a general modulator of the release of reactive oxygen species by phagocytes and is irrespective of phagocytic cellular type, or species from which the cells were derived.

Chromatography, High Pressure Liquid↗
PubMed1993

Predictors of early saphenous vein aortocoronary bypass graft occlusion. The GESIC Study Group.

To identify factors determining early saphenous vein aortocoronary bypass occlusion, we analyzed the data base of the GESIC study, a trial comparing antiplatelet drug regimens that included 927 patients with 1,854 saphenous vein grafts. The univariate analysis showed female sex (p < 0.0097), obesity (p < 0.001), rest angina (p < 0.0026), history of congestive heart failure (p < 0.037), the revascularized artery (p < 0.0001), the quality of distal bed (p < 0.00001), the diameter of the grafted vessel (p < 0.00001), the lack of antiaggregant treatment (p < 0.017), and a nonsequential technique (p < 0.0002) as predictors of early (28 days) graft occlusion. In the multivariate analysis the last five variables were independent predictors. Using the two preoperative variables, it was possible to identify groups at different risk; the occlusion rate ranged between 8.79% (left anterior descending coronary artery and good distal vessel) and 27.58% (right coronary artery or left circumflex coronary artery and poor distal vessel). The combination of three variables (grafted vessel, artery diameter, and antiaggregant treatment) also allowed identification of different risk groups; the occlusion rate ranged between 3.5% and 63.1%. Thus, it is possible to anticipate the risk of graft occlusion in patients undergoing coronary artery bypass grafting, which may help in the selection of both patients and antithrombotic treatment.

Aged↗
PubMed1993

Is the antioxidant, anti-inflammatory putative new vitamin, PQQ, involved with nitric oxide in bone metabolism?

Our laboratory recently isolated free PQQ (2,7,9-tricarboxy-pyrroloquinoline quinone, methoxatin), a bacterial redox cofactor, from red cells, neutrophils, serum and milk and found free PQQ in CSF, synovial fluid and bile. The metabolism and functions of PQQ and ascorbate may be coupled. Physiologically, free PQQ catalyzes dioxygen-superoxide interconversion, and participates in both superoxide generation (respiratory burst) and scavenging (cell protection). Using a labeled aromatic o-diamine, superoxide formation by activated neutrophils was inhibited and the labeled phenazine adduct of PQQ could be isolated from the inhibited cells (Karnovsky et al., 1992). PQQ may convert xanthine oxidase to xanthine dehydrogenase (XD) and could be the physiological coenzyme of XD. PQQ plus copper, form a potent amine-oxidizing system. Shah et al., 1992 found that PQQ-Cu2+ catalyzes the oxidation of epsilon-amino groups in collagen and elastin. Rucker's lab (Smidt et al., 1991) has found that PQQ may be a vitamin for mouse pups. Watanabe et al., 1988 and Nishigori et al., 1989, showed that injected PQQ protects animals against oxidative stress injury. PQQ's in vivo antioxidant action, spares reduced glutathione. PQQ, as an actively transported organic anion, concentrates in cells. In other experiments (Aizenman et al., 1992), PQQ protected neurons against the neurotoxin action of the glutamate-receptor against NMDA. We shall consider possible roles for PQQ in the biosynthesis of nitric oxide (NO, endothelium-derived relaxing factor, EDRF) from L-arginine and in NO removal by superoxide. NO has now been linked to the inhibition of osteoclastic bone resorption.

Animals↗
PubMed1992

Hydrocortisone anaphylaxis: a new case report.

In this report we describe a case of a nonatopic patient who developed an anaphylactoid reaction immediately after receiving intravenous hydrocortisone. The patient recovered after reanimation techniques and intravenous administration of atropine, epinephrine and plasma expanders. Although allergic reactions to corticosteroids appear to be rare there are a few case reports in the literature. This case is presented to draw the attention of clinicians to the occasional hazard of intravenous corticosteroid preparations, specially hydrocortisone.

Anaphylaxis↗
PubMed1992

[Prognostic predictors of left ventricular aneurysm after myocardial infarct].

We have prospectively studied the prevalence and natural history of left ventricular aneurysm in a series of 386 consecutive male patients with an acute myocardial infarction. Definition of aneurysm was angiographic and based on a single plane left ventricular angiogram. There were 52 (13.5%) left ventricular aneurysms. Mean follow-up was 57 +/- 29 months (range 40-100). During this interval, 15 of the 52 patients died (29%). Univariate analysis showed that survivors had at admission significantly less prevalence of hypercholesterolemia, previous myocardial infarction and intraventricular conduction disturbances. In addition, their Killip class was better (p less than 0.02) and the left ventricular ejection fraction was higher (p less than 0.002). The survival rates of patients with ventricular aneurysm at 1, 3 and 5 years were 88, 82 and 75%, respectively. Left ventricular end diastolic pressure, number of diseased vessels and the presence of bifascicular block were independent predictors of survival. Stratification of patients according to the risk factors allowed identification of patients with excellent prognosis (those with left ventricular end diastolic pressure less than 20 mmHg, single vessel disease and no conduction disturbances) and others with high risk of death (left ventricular end diastolic pressure greater than or equal to 20 mmHg, three vessel disease and bifascicular block). Survival at 5 years for these two groups were 81% vs 50%, respectively (p less than 0.005).

Heart Aneurysm↗
PubMed1991

Specific detection of quinoproteins by redox-cycling staining.

Quinones and related quinonoid substances catalyze redox cycling at an alkaline pH in the presence of excess glycine as reductant. With nitroblue tetrazolium and oxygen present there is concomitant reduction of the tetrazolium to formazan. This property of quinonoid compounds is used for the specific staining of quinoproteins, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotted onto nitrocellulose. The dopa-containing vitelline proteins and the 6-hydroxydopa-containing bovine serum amine oxidase are stained with the nitroblue tetrazolium/glycinate reagent. Also, the mammalian quinoproteins, diamine oxidase and lysyl oxidase, purported to contain pyrroloquinoline quinone, tested positive in this procedure. No quinonoid components were detected in three putative pyrroloquinoline quinone-containing quinoproteins, dopamine beta-hydroxylase, lipoxygenase, and peptidylglycine-amidating monoxygenase. Redox-cycling staining therefore confirms the presence of covalently bound quinones in the copper-dependent amine oxidases, but not in two putative quinoprotein oxygenases. Clarification of the biological significance of quinolation should be facilitated by identification of quinoproteins using this approach.

Animals↗
PubMed1991

Biosynthesis and secretion of rat salivary proteins by Xenopus laevis oocytes.

Xenopus laevis oocytes injected with poly (A+) RNA isolated from rat parotid and submandibular glands synthesize and secrete salivary proteins. Amylase was identified in the media of cultured oocytes injected with rat parotid mRNA by size and immunoprecipitation with anti-human amylase serum. Secretion of the salivary proteins was detectable in the medium eight h following the parotid mRNA injection and continued in a time-dependent fashion for up to 96 h. In contrast to rat parotid slices in culture, which demonstrate a regulated pathway of secretion highly responsive to the secretagogue isoproterenol, secretion of salivary proteins by oocytes did not respond to the stimulation by isoproterenol. Though parotid mRNA is presumed to contain the templates encoding the regulated pathway of secretion, reconstitution of this pathway of secretion in oocytes was not observed in our experiments. Since Xenopus laevis oocytes secrete constitutively significant amounts of proteins when injected with salivary gland mRNA, they are a useful biological system for the analysis of secretion, processing, and function of salivary proteins.

Animals↗
PubMed1991

[Late recurrence in pheochromocytoma: need for a long term follow up].

Pheochromocytoma is a catecholamine secreting tumor which has been traditionally considered as a potentially curable cause of hypertension. However, the existence of malignant pheochromocytomas as well as the problem which the recurrence of benign pheochromocytomas present, make the eradication of this tumor more complicated. It is note worthy the difficulty in localizing recurrent pheochromocytomas as well as the prognostic interest that presents the possibility of differentiating between recurrence and metastasis. We refer the case of a female patient suffering benign adrenal pheochromocytoma, who, 10 years it was surgically removed, presented a relapse of symptoms, and was diagnosed of extra-adrenal pheochromocytoma, in Zuckerkandl organ. The importance of performing a long term post-surgical follow-up in these patients is stressed.

Adrenal Gland Neoplasms↗
PubMed1990

Soybean lipoxygenase-1 is not a quinoprotein.

Soybean lipoxygenase-1 was reinvestigated with respect to its quinoprotein nature. It has been reported previously that soybean lipoxygenase-1 contains pyrroloquinoline quinone as the organic cofactor. Because spectroscopic data were found to be inconsistent with the evidence presented in [1], we sought to reproduce the published data by carefully following the procedures described in [1] and supplementing them with new analytical results. The combined data lead us to conclude that soybean lipoxygenase-1 is not a quinoprotein.

Coenzymes↗