Damjanović, Ana

Djurovic, M. N.

Draskovic, D.

Jadranin, Milka

Juranic, Z.

Juranić, Ivan O.

Pudar, G.

Rašković, Sanvila S.

Tešević, Vele

Trifunović, Jelena

Immunology

Cherry - Repository of the Faculty of Chemistry;  University of Belgrade

LC/DAD analysis of serum biogenic amines in patients with diabetes mellitus, chronic urticaria and Hashimoto's thyroiditis

Juranić, Z.

Faculty of Chemistry Repository - Cherry

Trifunovic, J.

Damjanovic, A.

Raskovic, S.

Juranić, Ivan

Juranić, Zorica

Central Repository of the Institute of Chemistry, Technology and Metallurgy (CER)

P1588LC/DAD analysis of serum biogenic amines in patients withdiabetes mellitus, chronic urticaria and Hashimoto’s thyroiditisJ. Trifunovic,* M. Jadranin, A. Damjanovic, S. Raskovic,§M. N. Djurovic,– D. Draskovic,– G. Pudar,** V. Tesevic,I. Juranic & Z. Juranic*Organic Chemistry, Faculty of Chemistry, Innovative Center, Universityof Belgrade, Belgrade, Serbia, Center for Chemistry, Institute forChemistry Technology and Metallurgy, University of Belgrade, Belgrade,Serbia, Experimental Oncology, Institute for Oncology and Radiology ofSerbia, Belgrade, Serbia, §Immunology and Allergology, Institute ofImmunology and Allergology Clinical Center of Serbia, Belgrade, Serbia,–Endocrinology, Institute of Endocrinology Clinical Center of Serbia,Belgrade, Serbia, **Endocrinology Diabetes and Metabolism Diseases,University Clinical Center ‘‘Zvezdara’’, Belgrade, Serbia, OrganicChemistry, Faculty of Chemistry University of Belgrade, Belgrade, Serbia,Organic Chemistry, Institute for Chemistry Technology and Metallurgy,University of Belgrade, Belgrade, SerbiaPurpose/Objective: The biogenic amines putrescine (Put), histamine(His), spermidine (Spd), N-acetyl putrescine (NAP), N-acetyl sper-midine (NAS), dopamine (Dop), epinephrine (Epi), norepinephrine(NE) and spermine (Spm) are a group of naturally occurring com-pounds exerting a large number of biological effects.This study was commenced to elucidate the role of biogenic aminesas possible diagnostic markers for three autoimmune diseases: Diabetesmellitus, Chronic urticaria, and Hashimoto’s thyroiditis.Materials and methods: This study involved 20 patients with Diabetesmellitus, 20 patients with Chronic urticaria, eight patients withHashimoto’s thyroiditis, and 20 healthy volunteers. We precipitatedserum proteins using 0.4 M HClO4. At pH 8.0 we performedderivatization with dansyl-chloride. 50 ll of prepared serum sampleswere injected into LC/DAD, in conditions of gradient elution, on C18column. Commercially available Put, His, Spd, NAP, NAS, Dop, Epi,NE, and Spm were dissolved in different concentrations in ultra purewater; treated in the same way as serum samples and injected into LC.Calibration curves were made by plotting peak area values against therespective concentrations of standards. The qualitative analysis wasdone using the method of retention time, and quantitative analysisusing external calibration. The recovery study was carried out usingreal serum sample from healthy control, by spiking techniques.Results: Retention times were 6.6 min for NAS, 8.8 min for NAP,9.1 min for Put, 10.1 min for His, 13.2 min for Spd, 13.9 min for NE,14.7 min for Epi, 14.9 min for Dop, and 15.4 min for Spm,respectively. Obtained data showed excellent linearity of calibrationcurves for Put, His, Spd, NAP, NAS, Dop, Epi, NE, and Spm.Compared to controls, His levels in Diabetes mellitus patients werestatistically higher; in Chronic urticaria patients levels of Put and Hiswere lower; Spd, NE and Epi levels were enhanced; Put was statisticallylower and Spd higher in Hashimoto’s thyroiditis patients. Chronicurticaria patients were the only in whose serum NE was found. NAP,NAS, Dop, and Spm were under limits of detection.Conclusions: Preliminary results from this study showed diversedistribution of investigated biogenic amines indicating differentactivation of metabolic pathways controlling biogenic amine biosyn-thesis and degradations in analyzed autoimmune diseases.P1589Levels of inﬂammatory mediators in gingival crevicular ﬂuid frompatients with periodontal disease before and after treatmentP. M. Alberga,* A. Romero, M. Correnti & L. Escalona*Postgrado en Ciencias Fisiolo´gicas, Instituto de Medicina Experimental,Caracas, Venezuela, Facultad de Odontologı´a, Instituto de InvestigacionOdontologica, Caracas, VenezuelaPurpose/Objective: The purpose of the present research was todetermine the levels of IL-1a, IL-1b, TNF-a, MMP-3 and MMP-8 ingingival crevicular fluid (GCF) of subjects with chronic periodontitisbefore and after non surgical treatment.Materials and methods: Clinical measurements were carried out in 11patients diagnosed with chronic periodontitis and 11 periodontallyhealthy controls. The clinical indexes evaluated were: gingival index(GI), plaque index (PI), bleeding on probing (BOP), probing depth(PD) and attachment loss (AL). The measurements were taken at sixsites per tooth in all teeth in each subject. GCF samples were takenfrom one tooth per quadrant, and the levels of these mediators weremeasured using an ELISA test.Results: A statistically significant difference (P < 0.05) was observed inall the clinical parameters between patients and control group before andafter periodontal treatment. Correlations between levels of mediatorswith the clinical parameters were not observed. Statistically significantdifferences were found between patients and control group in relationwith levels of inflammatory mediators (P < 0.05) before and aftertreatment. The levels of IL-1a, IL1-b, TNF-a, MMP-3 and MMP-8decreased in 44.2%, 48.35%, 53.28%, 62.16% y 34.03% respectively.Conclusions: Periodontal therapy reduced the levels of the inflam-matory mediators evaluated in this study, which were significantlyassociated with the severity of periodontal disease. This research wassupported by CDCH PG: 10-00-7070-2007.P1590Measuring autoantibodies against IL-17F and IL-22 in autoimmunepolyedocrine sydnrome type IB. E. V. Oftedal,* A. Meager, E. S. Husebye & A. S. B. Wolff**Endocrinology, Institute of Medicine, Bergen, Norway, Biotherapeutics,The National Institute for Biological Standards and Control, SouthMimms, UK, Endocrinology, Institute of Medicine, Haukeland Univer-sity Hospital, Bergen, NorwayPurpose/Objective: Patients with autoimmune polyendocrine syn-drome type I (APS I) have at least two of the three disease componentsadrenal insufficiency, hypoparathyroidism and chronic mucocutane-ous candidiasis. Various other organ-specific autoimmune manifesta-tions are common, as well as a number of ectodermal symptoms. Theunderlying cause of APS I is mutations in the gene encoding theautoimmune regulator protein AIRE. Deficiency of this protein leadsto loss of immunologic tolerance and release of autoreactive T-cellsfrom thymus into the periphery.Patients frequently develop high titers of autoantibodies againstmolecular targets in their affected organs. The pathological role ofthese antibodies is unknown, but they have become important markersfor APS I and other autoimmune conditions. Autoantibodies againstinterleukin (IL) -17A, IL-17F and IL-22 have recently been described inpatients with APS I, and their presence is reported to be highlycorrelated to chronic mucocutaneous candidiasis (CMC). The aim ofthis study was to develop a robust high-throughput radioligandbinding assays (RLBA) measuring IL-17F and IL-22 antibodies, and tocompare them with current enzyme-linked immunosorbent assays(ELISA) of IL-17F and IL-22; moreover to correlate the presence ofthese antibodies to the presence of CMC.Materials and methods: A total of five RLBAs were developed basedon IL-17F and IL-22 monomers and homo- or hetero dimers. As theseAbstracts 2012 The Author(s)  2012 Blackwell Publishing Ltd, Immunology, 137 (Suppl. 1), 185772673

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LC/DAD analysis of serum biogenic amines in patients with diabetes mellitus, chronic urticaria and Hashimoto's thyroiditis

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Cherry - Repository of the Faculty of Chemistry; University of Belgrade

Faculty of Chemistry Repository - Cherry

Central Repository of the Institute of Chemistry, Technology and Metallurgy (CER)