Enzyme Diamine Oxidase Research Articles

W1743 Intestinal Fat Absorption Is Associated with the Release of Histamine and Its Degrading Enzyme Diamine Oxidase

W1743 Intestinal Fat Absorption Is Associated with the Release of Histamine and Its Degrading Enzyme Diamine Oxidase

P236 - MEFV mutations modify the disease severity of inflammatory bowel disease

binding protein 1, ABP1; EC 1.4.3.6). ABP1 enzyme activity in bowel mucosa of patients with IBD is 50% lower than in healthy individuals. The reason for this decrease remains unknown, but it may be due in part to polymorphisms in the ABP1 gene. The present study investigated whether a common single nucleotide polymorphism (SNP, refSNP ID: rs1049793), which is located in exon 3 C2029G) and cause amino acid substitutions (His645Asp) in the ABP1 enzyme, is related to CD. Aims: To analyse the association between the presence of a non-synonymous single nucleotide polymorphism at diamine oxidase gene and the risk of developing Crohn’s disease (CD) and to analyse its influence on the clinical course of these patients. Methods: In this prospective, case-control study, 210 unrelated Caucasian consecutive CD patients were recruited at the Inflammatory Bowel Disease Unit of a single tertiary centre (Hospital Clinico San Carlos) in Madrid, Spain. All patients were phenotyped and followed up for a median time of 8.7 years (range 4.1 14.5 years). A total of 261 healthy volunteers from the same geographic area were also recruited and matched with patients. Both cases and controls were analysed for the presence of His645Asp amino acid substitutions in the diamine oxidase enzyme, using amplification-restriction procedures. The protocol was approved by the Ethics Committee of the Hospital and all patients and controls gave informed consent before inclusion in the study. Results: No significant differences were found in the distribution of ABP1 alleles between CD patients and healthy volunteers [for variant alleles = OR 1.15 (95%CI 0.86 1.55)]. The distribution of ABP1 genotypes did not differ when patients were subdivided according to gender, mean age at diagnosis, mean duration of the disease, family history of IBD, smoking habit, previous appendectomy or tonsillectomy, Montreal classification, perianal CD, extraintestinal manifestations of CD (cutaneous, articular, ocular and hepatic) and severity, i.e. need for immunosuppressive therapies, biological treatments and/or surgery. Conclusion: Our results suggest that the His645Asp polymorphism of the histamine metabolising enzyme ABP1 may not be related to the risk of developing CD. Moreover, this gene does not seem to play a role in disease activity.

Transcriptomic Analysis of Polyamine-Related Genes and Polyamine Levels in Placenta, Yolk Sac and Fetus During the Second Half of Mouse Pregnancy

In mammals, polyamines are essential for the maintenance of cell growth. Although early studies reported the highest values of mammalian ornithine decarboxylase (ODC) activity, a key enzyme in polyamine biosynthesis, in rodent placenta, the role of this enzyme in the second half of rodent pregnancy is still controversial. In order to get new insights on polyamine metabolism during this period of pregnancy, we studied polyamine levels, ODC expression and activity and transcript profile of different polyamine-related genes in mouse placenta, fetus and yolk sac. Results indicated that ODC activity and protein levels were higher in placenta than in fetus and yolk sac, especially in the labyrinth, although no correlation between ODC activity and polyamine levels were observed. The half-life of placental ODC (∼190 min) was also higher than the fetal one (∼24 min). Messenger RNAs of all biosynthetic and retroconversion enzymes of polyamine metabolism were present in the three gestational compartments analyzed, as well as those of antizymes 1 and 2 and antizyme inhibitor 1. However, no expression of antizyme 3 and antizyme inhibitor 2 was detected. The catabolic enzyme diamine oxidase was expressed only in the maternal part of placenta but not in the fetal part or in the fetus. The expansion of polyamine pools in the fetus was markedly higher than in placenta, in spite of its lower biosynthetic activity. Our results suggest that the elevated polyamine biosynthetic activity of mouse placenta is required to satisfy the high demand of polyamines required by the growing fetus, during the later period of pregnancy.

Analysis of a non-synonymous single nucleotide polymorphism of the human diamine oxidase gene (ref. SNP ID: rs1049793) in patients with Crohn's disease

Objective. To analyse the possible influence of a non-synonymous single nucleotide polymorphism (SNP) of the histamine-degrading enzyme diamine oxidase (DAO) on genetic susceptibility to Crohn's disease (CD). Material and methods. In this prospective, case-control study, 210 unrelated Caucasian consecutive CD patients were recruited at the Inflammatory Bowel Disease Unit of a single tertiary centre (Hospital Clínico San Carlos) in Madrid, Spain. A total of 261 healthy volunteers from the same geographic area were also recruited and matched with patients. Both cases and controls were analysed for the presence of a non-synonymous SNP (rs1049793) of DAO using amplification-restriction procedures of the genotype obtained in a blood sample. Results. No significant differences were found in the distribution of carriers of the non-synonymous SNP of DAO between CD patients and controls (OR 1.2 (95% CI 0.9–1.6; p=0.3)). Nor were any differences found between carriers and non-carriers of the non-synonymous SNP in demographic characteristics, phenotypes, complications or treatment of CD. Conclusions. The study of a non-synonymous SNP (rs1049793) of DAO does not seem to be of use in assessing susceptibility to CD, either as a marker of disease activity or as a marker of clinical behaviour in patients with the disease.

Effects of histamine and diamine oxidase activities on pregnancy: a critical review

Histamine has been assumed to contribute to embryo-uterine interactions due to its vasoactive, differentiation and growth-promoting properties. However, its exact functions in pregnancy are unclear. The histamine-degrading enzyme diamine oxidase (DAO) is produced in high amounts by the placenta and has been supposed to act as a metabolic barrier to prevent excessive entry of bioactive histamine from the placenta into the maternal or fetal circulation. The literature available on PubMed published in English between 1910 and 2008 has been searched using the isolated and combined key words histamine, diamine oxidase, pregnancy, placenta, endometrium, miscarriage, implantation, pre-eclampsia, intrauterine growth retardation, diabetes and embryonic histamine-releasing factor (EHRF). High expression of the histamine-producing enzyme histidine decarboxylase in the placenta, histamine receptors at the feto-maternal interface and the existence of an EHRF suggest a physiological role of histamine during gestation. The balance between histamine and DAO seems to be crucial for an uncomplicated course of pregnancy. Reduced DAO activities have been found in multiple heterogeneous complications of pregnancy such as diabetes, threatened and missed abortion and trophoblastic disorders. Whether women with histamine intolerance suffer from more complicated pregnancies and higher abortion rates due to impaired DAO activities and if low DAO levels or genetic modifications in the DAO gene might therefore represent a prognostic factor for a higher risk of abortion, has not been investigated yet. Low activities of the histamine-degrading enzyme DAO might indicate high-risk pregnancies, although high intra- and interindividual variations limit its value as a screening tool.

Highly Sensitive Determination of DAO Activity by Oxidation of a Luminescence Reagent

A highly sensitive method for measuring the activity of the enzyme diamine oxidase (DAO) independent of the type of substrate is described. The principle of the assay is to determine the amount hydrogen peroxide generated as a reaction product during oxidation of diamines by DAO. PSatto, a highly sensitive luminescence reagent, was used to generate a signal depending on the hydrogen peroxide concentration based on the action of horseradish peroxidase. DAO is specifically captured from a sample by an antibody immobilized to microwell plates, and the substrate is added to the bound enzyme. Various diamines were used as substrates; the peroxide produced is directly proportional to the amount of DAO bound to the specific antibodies. With this very sensitive method, it is possible to detect pmol amounts of generated hydrogen peroxide in plasma matrix corresponding to the biological activity of DAO.

Improved analytical sensitivity reveals the occurrence of gender-related variability in diamine oxidase enzyme activity in healthy individuals

Objectives: Serum diamine oxidase (DAO; EC 1.4.3.6) activity is often employed as a clinical indicator of the integrity of intestinal mucosa. However, interindividual variation in enzyme activity and reference values for healthy women and men have not been studied. Design and methods: DAO activity was measured by using cadaverine coupled to O-dianisidine oxidation in 50 healthy individuals. Results: The mean activity was 7.59 ± 3.67 U/L for women and 2.38 ± 0.71 U/L for men ( p < 0.001). Conclusions: Gender is a major determinant for DAO activity in healthy subjects.

Histaminintoleranz: oft verkannt - einfach zu diagnostizieren

In contrast to food allergies, histamine intolerance is not mediated by IgE. The intensity of the symptoms of histamine intolerance increases with the amount of histamine ingested. Histamine is produced by the human body and by intestinal bacteria such as Morganella morganii. It is ingested with histamine-rich foods or liberated from mast cells by certain drugs. The enzymes diamine oxidase (DAO) and histamine N-methyltransferase are responsible for the degradation of histamine in the human body. In histamine intolerant individuals the enzyme diamine oxidase is not produced in sufficient amounts. Hence, histamine accumulates in the body and leads to symptoms such as shortness of breath, hypotension, flushing, urticaria, nausea, vomiting, headache and diarrhoea. In order to diagnose histamine intolerance, the presence of diamine oxidase in blood and Morganella morganii in fecal samples can be analysed. In case histamine intolerance is diagnosed, it can be met by a histamine-low diet. Antihistaminics and a diamine oxidase-supplement can be administered in acute cases or if Morganella morganii is the source of histamine production.

An amperometric biosensor for the rapid assessment of histamine level in tiger prawn ( Penaeus monodon) spoilage

Histamine could accumulate in seafood when bacteria spoilage commenced and caused histamine poisoning without altering the fish normal appearance and odor. Therefore, a histamine biosensor using immobilized enzyme diamine oxidase (DAO) has been developed for the rapid monitoring of the histamine levels in tiger prawn ( Penaeus monodon). The histamine biosensor had a response time of <1 min and optimum pH of operation was 7.4 with reproducibility and repeatability ( n = 5) of 4.87% and 5.26% relative standard deviations (RSD) respectively. Recoveries ranging from 93.11% to 100.58% were obtained for histamine spiked at levels from 5 to 20 ppm. The variation in histamine levels of some tiger prawn samples after a 5-h exposure at temperature of 30 °C ± 2 were studied using the histamine biosensor and the results were comparable to histamine levels determined by an HPLC method. The two methods showed a linear correlation with R 2 = 0.9612 ( Y = 0.9164 x + 5.58). The limit of detection was 0.65 ppm of histamine, which is below the indicator level of 50 ppm established by USA FDA. The reusable biosensor is simple and can be used for direct histamine determination without further sample pretreatment, and is suitable for the routine analysis of histamine in tiger prawns to monitor spoilage.

Altered growth and polyamine catabolism following exposure of the chocolate spot pathogen Botrytis fabae to the essential oil of Ocimum basilicum

Biomass of the fungal pathogen Botrytis fabae in liquid culture amended with two chemotypes of the essential oil of basil, Ocimum basilicum, was reduced significantly at concentrations of 50 ppm or less. The methyl chavicol chemotype oil increased the activity of the polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC), but polyamine concentrations were not significantly altered. In contrast, the linalol chemotype oil decreased AdoMetDC activity in B. fabae, although again polyamine concentrations were not altered significantly. However activities of the polyamine catabolic enzymes diamine oxidase (DAO) and polyamine oxidase (PAO) were increased significantly in B. fabae grown in the presence of the essential oil of the two chemotypes. It is suggested that the elevated activities of DAO and PAO may be responsible, in part, for the antifungal effects of the basil oil, possibly via the generation of hydrogen peroxide and the subsequent triggering of programmed cell death.

Altered growth and polyamine catabolism following exposure of the chocolate spot pathogen Botrytis fabae to the essential oil of Ocimum basilicum

Biomass of the fungal pathogen Botrytis fabae in liquid culture amended with two chemotypes of the essential oil of basil, Ocimum basilicum, was reduced significantly at concentrations of 50 ppm or less. The methyl chavicol chemotype oil increased the activity of the polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC), but polyamine concentrations were not significantly altered. In contrast, the linalol chemotype oil decreased AdoMetDC activity in B. fabae, although again polyamine concentrations were not altered significantly. However activities of the polyamine catabolic enzymes diamine oxidase (DAO) and polyamine oxidase (PAO) were increased significantly in B. fabae grown in the presence of the essential oil of the two chemotypes. It is suggested that the elevated activities of DAO and PAO may be responsible, in part, for the antifungal effects of the basil oil, possibly via the generation of hydrogen peroxide and the subsequent triggering of programmed cell death.

Optimierter Radioextraktionsassay zur quantitativen Bestimmung der Aktivität von Diaminooxidase (DAO) in humanem Serum und Plasma

Patients intolerant to histamine-containing foods (histamine intolerance) frequently suffer from diminished histamine degradation based on deficiency or reduced activity of the enzyme diamine oxidase (DAO). Here, we present an easy-to-use, commercially available test kit for the quantification of DAO activity in blood. Radiolabeled putrescine is added as a substrate to 100 μl of plasma or serum. DAO converts one amino group to an aldehyde. The resulting molecule spontaneously reacts to cyclic Δ 1 -pyrroline which can be extracted selectively by ethylacetate. With an optimized buffer system, we could improve the yield of extractable reactants four-fold compared to previous systems. A calibration curve with DAO from porcine kidney is used for quantification. Intraassay variation is < 10% (3.5 - 14%,n = 20), interassay variation is < 15% (4 - 21%, n = 10). Mean recovery of DAO added to samples was 104% (86 - 123%, n = 20). Results are available within three hours. DAO activity in samples was stable at 4 °C for one week, storage at -20 °C with repeated freeze cycles did not affect activity. This assay is a reliable tool for the determination of blood DAO activity and is a helpful tool in the diagnosis of non-allergic food hypersensitivities. Due to the good stability of DAO, blood samples may be drawn in the doctors office and shipped to the diagnostic lab.

Local and systemic changes in arginine decarboxylase activity, putrescine levels and putrescine catabolism in wounded oilseed rape

* Here we report the effect of mechanical wounding on putrescine biosynthesis and catabolism in oilseed rape (Brassica napus ssp. oleifera). * The lamina of first leaves was wounded by crushing with forceps, and first and second leaves were harvested at various intervals over a 24 h period. Levels of free polyamines were measured and activities of enzymes of polyamine biosynthesis and catabolism were assayed in the harvested tissue. * Mechanical wounding of the first leaves led to significant, but transient, increases in arginine decarboxylase (ADC) activity and levels of free putrescine in the wounded first leaf and in unwounded second leaves. The increased putrescine appeared to be the result of a combination of increased ADC activity, coupled with reduced putrescine catabolism, as activity of the oxidative enzyme diamine oxidase was significantly reduced following wounding, both locally and systemically. * The role of the increased free putrescine in the wound response of oilseed rape is not known, although the possibility that it is used to form putrescine conjugates is worthy of further investigation.

611 Introduction of specificity into cytotoxic drugs and improvement of therapeutic index by kinase-mediated trapping

The polyamines putrescine, spermidine, and spermine are oxidized by the enzyme diamine oxidase to form the corresponding aldehyde derivatives. These aldehydes have been shown to undergo a variety of spontaneous reactions, some of which result in polycationic addition compounds. We have chemically synthesized some spermine-derived polycations by reaction with the dialdehyde glutaraldehyde followed by reduction of the resulting Schiff base with sodium borohydride. Their migration on ion exchange and gel filtration columns was consistent with the formation of polycations with properties similar to those reported for the spontaneous reaction products. When added to cultures of alloantigen or mitogen stimulated lymphocytes, these polycations were potent inhibitors of the incorporation of tritiated thymidine and blast cell formation. This inhibition was reversible, non-cytotoxic, and only apparent if the polycation was added early in the culture period. The concentration of polycation necessary to achieve 50% inhibition of the lymphocyte response decreased as the cationic nature relative to spermine increased.

Continuous administration of heparin in patients with deep vein thrombosis can increase plasma levels of diamine oxidase

Besides its anticoagulant activity, the sulfated polysaccharide heparin has numerous other biological effects. Especially the antiinflammatory and immunoregulatory properties of heparin may be associated with its ability to release the histamine-degrading enzyme diamine oxidase (DAO) from tissue-bound sites into the circulation. Whereas DAO activity is at the limits of detection in normal human plasma, the application of heparin leads to a significant increase of plasma DAO activity. However, previously, only the effect of bolus injection of unfractionated heparin (UFH) had been studied. To investigate DAO release during continuous heparin infusion, 28 patients with deep vein thrombosis (DVT) undergoing heparin therapy were analyzed. Whereas continuous heparin infusion did not lead to any increase of plasma DAO activity in 12 patients (43%), 6 patients (21%) showed a single elevated and 10 patients (36%) permanently elevated plasma DAO activity. The groups of patients exhibiting different DAO release responses did not differ in age, sex, body weight, concomitant diseases, heparin infusion rates, coagulation indices, location and extension of thrombosis, or clinical outcome. However, the rate of idiopathic DVT was significantly higher in the group of patients releasing DAO. This study shows, for the first time, that continuous heparin infusion can lead to DAO release and that individuals exhibit considerable differences in their release response. Although the significance of heparin-induced DAO release needs further clarification, our results indicate that postheparin plasma DAO activity could be an interesting parameter correlated with idiopathic DVT.

Histamine-degrading enzymes as cellular markers of acute small bowel allograft rejection

Abstract Intestinal histaminedegrading enzymes diamine oxidase (DAO) and histamine N-methyltransferase (HNMT) activities are relatively constant per individual and bowel segment, and they reflect the functional integrity of the intestinal mucosa. It was, therefore, hypothesised that a decrease in these enzymes could be indicative of acute rejection of an intestinal allograft. Enzymatic activities of DAO and HNMT were determined in mucosal biopsies of isogeneic (Lewis-to-Lewis, n=48) and allogeneic (Brown Norway-to-Lewis, n=48) heterotopic small bowel transplants in a rat model at various time periods. Allograft recipients were not given any immunosuppression. While no changes in enzyme activities were observed in isografts up to day 8 following transplantation, significantly reduced activities of both enzymes were found in all allografts 6–8 days after transplantation. Activities of both DAO and HNMT exhibited a strong negative correlation with the histological rejection score (P<0.01). We can conclude that DAO and HNMT activities in gut mucosa are reliable quantitative markers of acute intestinal allograft rejection in the rat that support histopathological analysis.

Histamine-degrading enzymes as cellular markers of acute small bowel allograft rejection.

Intestinal histamine-degrading enzymes diamine oxidase (DAO) and histamine N-methyltransferase (HNMT) activities are relatively constant per individual and bowel segment, and they reflect the functional integrity of the intestinal mucosa. It was, therefore, hypothesised that a decrease in these enzymes could be indicative of acute rejection of an intestinal allograft. Enzymatic activities of DAO and HNMT were determined in mucosal biopsies of isogeneic (Lewis-to-Lewis, n=48) and allogeneic (Brown Norway-to-Lewis, n=48) heterotopic small bowel transplants in a rat model at various time periods. Allograft recipients were not given any immunosuppression. While no changes in enzyme activities were observed in isografts up to day 8 following transplantation, significantly reduced activities of both enzymes were found in all allografts 6-8 days after transplantation. Activities of both DAO and HNMT exhibited a strong negative correlation with the histological rejection score ( P<0.01). We can conclude that DAO and HNMT activities in gut mucosa are reliable quantitative markers of acute intestinal allograft rejection in the rat that support histopathological analysis.

Pharmacological analysis of red-wine-induced migrainous headaches

We describe a series of experiments designed to investigate the mechanisms by which headaches can be triggered by red wine in a small minority of migraine patients. Some red wines are particularly potent releasers of serotonin from platelet stores, but these are no more effective as triggers of headache in sensitive patients. Both the selective 5-HT2A antagonist ketanserin and the non-selective 5-HT2ABC antagonist pizotifen blocked the majority of headaches, and we then thought the antihistamine properties of these two drugs might be important. In a third experiment, however, the H1 antagonist mepyramine did not convincingly antagonise the response to red wine. Plasma levels of the enzyme diamine oxidase, which metabolises histamine, were lower in all migraine patients, whether or not they were sensitive to red wine. The results reported here do not permit making firm conclusions; nevertheless it seems that different pharmacological receptors may be responsible in different patients.

Simultaneous purification of the histamine degrading enzymes diamine oxidase and histamine N-methyltransferase from the same tissue.

Simultaneous purification of the histamine degrading enzymes diamine oxidase and histamine N-methyltransferase from the same tissue.

Polyamine Metabolism in an Incompatible Interaction between Barley and the Powdery Mildew Fungus, Blumeria graminis f. sp. hordei

AbstractIn the incompatible interaction between the barley cultivar Hart (mlo allele) and the powdery mildew fungus Blumeria graminis DC. Speer f. sp. hordei Marchal, free spermidine and conjugated forms of putrescine and spermidine were found to accumulate 1–3 days following inoculation. The increase in free spermidine could be accounted for by increased activities of the biosynthetic enzymes ornithine decarboxylase (EC 4.1.1.17) and S‐adenosylmethionine decarboxylase (EC 4.1.1.50). Activities of the transferase enzymes putrescine hydroxycinnamoyl transferase (EC 2.3.1.‐) and tyramine feruloyl‐CoA transferase (EC 2.3.1.‐) were also increased in leaves of Hart inoculated with powdery mildew, as were activities of the polyamine catabolic enzymes diamine oxidase (DAO, EC 1.4.3.6) and polyamine oxidase (PAO, EC 1.4.3.4). The possible role of conjugated polyamines, and increases in DAO and PAO, in penetration based resistance to powdery mildew infection is discussed.