Characterization of recombinant human diamine oxidase (rhDAO)produced in Chinese Hamster Ovary (CHO) cells
Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells.
- Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria; Department of Clinical Pharmacology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.
- Department of Chemistry, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
- Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9, 1060 Vienna, Austria.
- Department of Clinical Pharmacology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.
- Polymun Scientific Immunbiologische Forschung GmbH, Donaustraße 99, 3400 Klosterneuburg, Austria.
- Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
- Department of Clinical Pharmacology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria. Electronic address: firstname.lastname@example.org.
Human diamine oxidase (hDAO) efficiently degrades polyamines and histamine. Reduced enzyme activities might cause complications during pregnancy and be involved in histamine intolerance. So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. Accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. Topaquinone (TPQ) cofactor content was determined using phenylhydrazine titration. Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. Mass spectrometric analysis was conducted to verify sequence integrity of the protein and to assess the glycosylation profile. Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. The substrate preference and kinetic parameters were in accordance with previous publications. The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions.
AOC1; Amiloride-sensitive amine oxidase; CHO; Chinese Hamster Ovary; DAO; Recombinant human diamine oxidase
Association of diamine oxidase and histamine N-methyltransferase polymorphisms with presence of migraine in a group of Mexican mothers of children with allergies
Association of diamine oxidase and histamine N-methyltransferase polymorphisms with presence of migraine in a group of Mexican mothers of children with allergies.
Low histamine metabolism has been suggested to play a role in the pathogenesis of allergy and migraine. We investigated the possible association between 2 single-nucleotide polymorphisms (SNP), C314T HNMT and C2029G DAO, and the presence and severity of migraine and migraine-related disability.
Materials and methods
We studied the frequency of C314T HNMT and C2029G DAO allelic variants in 162 mothers of children with allergies (80 with migraine and 82 without) using a TaqMan-based qPCR Assay and a case–control model. We conducted a logistic regression analysis to examine the association between migraine and the allelic and haplotype variants.
Mutant C2029G DAO SNP was found significantly more frequently in the group of women with migraine than in controls (OR, 1.6; 95% CI, 1.1–2.1). No significant differences were found in frequencies of genotypes or alleles in the case of C314T HNMT SNP. Both mutated alleles were associated with migraine-related disability. Coexistence of alleles for both SNPs (haplotypes) showed a strong association with migraine. Haplotypes containing both mutated alleles (either heterozygous or homozygous) were very strongly associated with MIDAS grade IV migraine (OR, 45.0; 95% CI, 5.2–358). This suggests that mutant alleles of C314T for histamine N-methyltransferase (HNMT) and C2029G for diamine oxidase (DAO) polymorphisms may interact in a way that increases the risk and impact of migraine.
We suggest a synergistic association between HNMT and DAO functional polymorphisms and migraine; this hypothesis must be further confirmed by larger studies. However, the characteristics and ethnic differences between analysed populations should be considered when interpreting the results.
Diamiamine oxidase (DAO) activity in serum patients with migraine
Latorre-Moratalla, M.L.a,Comas;Basté, O.a, Izquierdo-Casas, J.b,c, Soler-Singla, L.b,c, Lorente-Gascón, M.c, Duelo, A.d, & Vidal-Carou, M.C.a
a Department of Nutrition, Food Sciences and Gastronomy – XaRTA – INSA, Faculty of Pharmacy and Food Sciences, University of Barcelona.
b Department of Neurology, Hospital General de Catalunya.
c Department of Basic Sciences, Universitat Internacional de Catalunya.
d Department of Nutrition, Instituto Clínico del Déficit de DAO (ICDDAO)
Avinguda Prat de la Riba 171, 08921 Santa Coloma de Gramenet (Barcelona), Spain. email@example.com
Diamine oxidase levels in different chronic urticaria phenotypes.
Allergol Immunopathol (Madr). 2015 Nov-Dec;43(6):593-600. doi: 10.1016/j.aller.2015.01.009. Epub 2015 May 13.
Diamine oxidase (DAO) is a polyamine-degrading enzyme also implicated in histamine metabolism. Chronic urticaria (CU) has a wide spectrum of clinical presentations and causes. Anisakis sensitisation associated chronic urticaria (CU+) has been characterised as a phenotype with different clinical and immunological characteristics and possibly associated with previous acute parasitism. We aimed to analyse serum DAO levels in different CU phenotypes. We further analysed the possible association of DAO with fish eating habits.
We studied 35 CU+ patients and 39 non-sensitised CU patients (CU-) as well as 19 controls. We analysed fish-eating frequency as well as fish intake associated exacerbation of CU (FIAE) or gastro-intestinal complaints (GI). DAO levels were further analysed with respect to lymphoproliferative responses, cytokine and specific IgE production.
DAO levels were not different between CU and controls, but were significantly higher in CU+ than in CU-. CU+ patients with FIAE had lower DAO levels, but no differences were detected in patients with GI. DAO levels correlated positively with oily and canned fish consumption in CU-. In CU+, DAO levels correlated positively with specific Anisakis IgE, percentages of proliferation in Anisakis stimulated peripheral blood lymphocytes, serum IL-2 and IL-6, but correlated negatively with mitogen stimulated TGF-β in supernatants.
DAO levels in CU depend on fish-eating habits and in CU+ on the amount of specific IgE production. In the CU+ phenotype, lower levels of DAO predispose to urticaria exacerbation after fish intake, probably due to a relative insufficient enteric availability of this enzyme.
Copyright © 2014 SEICAP. Published by Elsevier Espana. All rights reserved.
Anisakis; Chronic urticaria; Cytokines; Diamine oxidase; Diet
Serum diamine oxidase activity as a predictor of gastrointestinal toxicity and malnutrition due to anticancer drugs
Serum diamine oxidase activity as a predictor of gastrointestinal toxicity and malnutrition due to anticancer drugs.
- Department of Gastroenterology and Oncology, Institute of Health Biosciences, University of Tokushima Graduate School, Tokushima, Japan.
- Department of General Medicine and Community Health Science, Institute of Health Biosciences, University of Tokushima Graduate School, Tokushima, Japan.
BACKGROUND AND AIM:
Objective evaluation of intestinal mucosal damage due to anticancer drugs is generally difficult. Serum diamine oxidase (DAO) activity is reported to reflect the integrity and maturity of the small intestinal mucosa. Therefore, we investigated whether serum DAO activity is an indicator of gastrointestinal toxicity or nutritional status in patients receiving chemotherapy.
We prospectively enrolled 20 patients with unresectable metastatic gastric cancer who received oral S-1 (80 mg/m(2) ) on days 1-14, and intravenous cisplatin (60 mg/m(2) ) and docetaxel (50 mg/m(2) ) on day 8 every 3 weeks. Serum DAO activity was measured by colorimetry. Gastrointestinal toxicity was evaluated by Common Toxicity Criteria for Adverse Events version 4.0. Endoscopic examination and biopsy of duodenal mucosa assessed mucosal damage. Malnutrition was evaluated by measuring serum total protein and albumin levels.
Serum DAO activity decreased step-by-step significantly during anticancer drug treatment and recovered after drug holidays. In all 14 patients who experienced diarrhea, serum DAO activity significantly decreased prior to diarrhea onset. Percent decrease in DAO activity was significantly correlated with severity of diarrhea. Significant correlation was observed between percent decrease in DAO activity and percent decrease in duodenal villus height or surface area from baseline. There were also significant correlations between percent decrease in serum DAO activity at day 14 and percent decrease in serum total protein or albumin levels at day 21 from baseline.
Serum DAO activity sensitively indicates gastrointestinal damage prior to symptom onset and can be a useful predictor of intestinal mucosal damage and nutritional status in patients receiving chemotherapy.
© 2015 Journal of Gastroenterology and Hepatology Foundation and Wiley Publishing Asia Pty Ltd.
anticancer drugs; diamine oxidase; diarrhea; gastrointestinal toxicity; malnutrition
Serum diamine oxidase activity in patients with histamine intolerance.
Intolerance to various foods, excluding bona fide coeliac disease and lactose intolerance, represents a growing cause of patient visits to allergy clinics.
Histamine intolerance is a long-known, multifaceted clinical condition triggered by histamine-rich foods and alcohol and/or by drugs that liberate histamine or block diamine oxidase (DAO), the main enzyme involved in the metabolism of ingested histamine. Histamine limitation diets impose complex, non-standardized restrictions that may severely impact the quality of life of patients.
We retrospectively evaluated 14 patients who visited allergy outpatient facilities in northern Italy with a negative diagnosis for IgE-mediated food hypersensitivity, coeliac disease, conditions related to gastric hypersecretion, and systemic nickel hypersensitivity, and who previously underwent a histamine limitation diet with benefits for their main symptoms. Serum diamine oxidase levels and the clinical response to diamine oxidase supplementation were investigated.
We found that 10 out of 14 patients had serum DAO activity <10 U/mL, which was the threshold suggested as a cutoff for probable histamine intolerance. Moreover, 13 out of 14 patients subjectively reported a benefit in at least one of the disturbances related to food intolerances following diamine oxidase supplementation. The mean value (± SD) of diamine oxidase activity in the cohort of patients with histamine intolerance symptoms was 7.04 ± 6.90 U/mL compared to 39.50 ± 18.16 U/mL in 34 healthy controls (P = 0.0031).
In patients with symptoms triggered by histamine-rich food, measuring the serum diamine oxidase activity can help identify subjects who can benefit from a histamine limitation diet and/or diamine oxidase supplementation.
Properly designed, controlled studies investigating histamine intolerance that include histamine provocation are indispensable for providing insights into the area of food intolerances, which are currently primarily managed with non-scientific approaches in Italy.
Histamine, histamine intoxication and intolerance.
Allergol Immunopathol (Madr). 2015 Sep-Oct;43(5):498-506. doi: 10.1016/j.aller.2015.05.001. Epub 2015 Aug 1.
- Comenius University in Bratislava, Jessenius Faculty of Medicine, Department of Pathophysiology, Mala Hora, 036 01 Martin, Slovakia.
- Comenius University in Bratislava, Jessenius Faculty of Medicine, Department of Pathophysiology, Mala Hora, 036 01 Martin, Slovakia. Electronic address: firstname.lastname@example.org.
Excessive accumulation of histamine in the body leads to miscellaneous symptoms mediated by its bond to corresponding receptors (H1-H4). Increased concentration of histamine in blood can occur in healthy individuals after ingestion of foods with high contents of histamine, leading to histamine intoxication. In individuals with histamine intolerance (HIT) ingestion of food with normal contents of histamine causes histamine-mediated symptoms. HIT is a pathological process, in which the enzymatic activity of histamine-degrading enzymes is decreased or inhibited and they are insufficient to inactivate histamine from food and to prevent its passage to blood-stream. Diagnosis of HIT is difficult. Multi-faced, non-specific clinical symptoms provoked by certain kinds of foods, beverages and drugs are often attributed to different diseases, such as allergy and food intolerance, mastocytosis, psychosomatic diseases, anorexia nervosa or adverse drug reactions. Correct diagnosis of HIT followed by therapy based on histamine-free diet and supplementation of diamine oxidase can improve patient’s quality of life.
Diamine oxidase; Histamine; Histamine intolerance; Scombroid poisoning
Diamine oxidase rs10156191 and rs2052129 variants are associated with the risk for migraine.
- Department of Biochemistry and Molecular Biology, University of Extremadura, Cáceres, Spain; Red de Investigación de reacciones adversas a alergenos y fármacos, Instituto de Salud Carlos III, Madrid, Spain.
Histamine has been implicated in the pathogenesis of migraine. We investigated the possible association between functional single nucleotide polymorphisms (SNPs) in the diamine oxidase gene (DAO; chromosome 7q36.1, involved in histamine metabolism) and the risk for migraine.
We studied the frequency of the rs2052129, rs10156191, rs1049742, and rs1049793 genotypes and allelic variants in 197 patients with migraine and 245 healthy controls using a TaqMan-based qPCR Assay.
The DAO SNP rs10156191, which is related to decreased DAO enzyme activity, is associated with the risk of developing migraine, particularly in women. The odds ratio (OR) for the defect allele positivity is 1.61 (95% confidence interval 1.31-2.37) for overall migraine patients and 2.08 (1.29-3.36) for women suffering from migraine. The association was not influenced by confounders such as the age at onset, the presence of aura, positivity of alcohol as a triggering factor, positive family history of aura, or family history of allergy. Multiple regression analyses did not confirm association with the rest of genetic factors.
Our findings, which should be framed as hypothesis generating, suggest that DAO genotypes and allelic variants are associated with the risk for migraine in Caucasian Spanish people, especially in women.
© 2015 American Headache Society.
biomarker; diamine oxidase gene; genetics; histamine; migraine; risk factor
Evaluation of Diamine Oxidase deficiency in patients with migraine.
Vidal Carou ● A1, Sabater Sales ● B2, Titus Albareda ● C3.
(A1) Nutrition and Bromatology Department of University of Barcelona.
(B2) MD in Pharmacy. Responsible for scientific research at Sabater Analysis (LABCO).
(C3) Honorary Member of the Spanish Society of Neurology, and Scientific Advisor of the Spanish Association of Headache Patients, AEPAC
Migraine is a complex, recurrent and disabling disease, but its etiopathogeny remains unknown, so it is necessary to find new approaches to the biochemical mediators of the disease. Histamine, one of the most important mediators, is mainly metabolized by DAO (Diamine Oxidase enzyme). Therefore, it is believed that a decrease in DAO activity may cause histamine excess, increasing the risk of suffering from clinical pictures such as migraine, among others.
To determine the rate of migraine patients presenting DAO deficiency compared to the observed rate in general population without migraine.
Target population consisted of migraine volunteers. Healthy controls were selected among volunteers of their micro social environment. Given a total sample size of 160 individuals, the studied aimed to determine DAO activity level in the participants by analytical determination using a validated procedure.
IHS diagnostic sheet, patient information sheet, informed consent form, 160 analytical tests to determine DAO, data collection logbook, documents for the registration in the data protection agency.
48.8 % of migraine patients presented values of much reduced DAO activity, and 46% of reduced activity. 95% of migraine patients experienced other symptoms related to histamine intolerance. The average value obtained for DAO activity is lower in the migraine group, with a significant difference (p=0.001).
The results confirm the scope of the research and show that DAO activity is significantly lower in migraine patients compared to controls.
Role of histamine and diamine oxidase enzyme in Multiple Sclerosis.
- Farokhi, M. Etemadifar, A. Rezaei, A. Amani, H. Jahanbani
Multiple Sclerosis and related disorders, November 2014, Volume 3, Issue 6, Page 746