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EICOSANOIDE - ihre Beteiligung an physiologischen und pathophysiologischen Prozessen �

TC Info+Veröffentlichungen-06-2005.doc 1

www.talkingcells.org

Allgemeine Informationen TalkingCells befasst sich mit der in vitro Untersuchung weißer Blutzellen, welche die Information über Erkrankungen und sogar deren Vorstufen 'erzählen', d.h. erkennen lassen.

Veränderte Stoffwechselvorgänge können Wegbereiter aber auch Ursache verschiedener Erkrankungen sein. Der Nachweis der hierbei von den Zellen gebildeten Stoffwechselprodukte (Mediatoren) ermöglicht die Erkennung pathologischen Prozesse und den Rückschluss auf mögliche pathobiochemische Mechanismen. Eicosanoide erwiesen sich hierbei als bedeutsame Informationsträger hinsichtlich normaler und veränderter Stoffwechselprozesse. Kürzlich wurden Funktionelle Eicosanoid Teste (z.B. LiPiDoC-AIT) für den klinischen Alltag entwickelt.

Einsatzgebiete Erkrankungen der oberen und unteren Atemwege

(z. B. Fließschnupfen, Nasennebenhöhlenentzündungen, Nasenpolypen, Asthma).

Hauterkrankungen (z.B. Nesselsucht

Veränderungen im Magen-Darm-Kanal (z.B. Magengeschwüre, entzündliche Darmerkrankungen)

sowie eine Reihe weiterer Erkrankungen

Praktischer Nutzen Der Funktionelle-Eicosanoid-Test

erleichtert und sichert die Diagnose bietet Therapiealternativen hat Bedeutung für die OP-Planung erkennt Neigungen zu Krankheiten bereits im Vorfeld, noch bevor

Beschwerden auftreten und die üblichen Untersuchungsmöglichkeiten fündig werden.

ist risiko-freie für den Patienten, ist einfach und zeitsparend (ambulante Blutabnahme) durchzuführen ist kostengünstig: - vermeidet zusätzliche Krankenhausaufenthalte - durch vorbeugende Maßnahmen lassen sich

Erkrankungen herauszögern oder sogar verhindern

unterstützt das Aufklärungsgespräch mit dem Patienten

http://www.talkingcells.org




Analgetika-Intoleranz: Das verkannte Krankheitsbild

Nur bei einem Teil der Patienten gibt sich die Analgetika-Intoleranz ohne spezielle Diagnostik zu erkennen: wenn stets nach Verabreichung von Analgetika die zuvor beschriebenen Erscheinungen auftreten - hier kann auf den Test verzichtet werden. Wo aber ein solcher Zusammenhang verborgen bleibt, hilft der Test bei der Aufklärung.

Dies ist der Fall, wenn die Beschwerden und Veränderungen erst mit großer Verzögerung eintreten, z.B. bei Nasenpolypen. Aber auch andere Stoffe können für die Symptome verantwortlich sein, z.B. Nahrungsmitteln oder Körperpflegeprodukte und Kosmetika.

Wurden bislang solche Zusammenhänge nicht erkannt, ist dies heute mit LiPiDoC-AIT möglich; galten die Betroffenen als chronisch progredient krank, so sind sie heute einer Prophylaxe und Therapie zugängig.

Erscheinungsformen der Analgetika-Intoleranz

Intoleranzen gegenüber schmerzlindernden oder schmerzstillenden Arzneimitteln (Analgetika) zählen nicht zu den Volkskrankheiten, obgleich sie weitverbreitete Erkrankungen und Beschwerden (z.B. Urtikaria, Polyposis nasi, Asthma bronchiale, Magen-Darm-Beschwerden) auslösen, die mit lebensbedrohlichen Reaktionen des Körpers einhergehen können. Wie bei vielen anderen Erkrankungen mit ähnlichem Hintergrund ist bei der Analgetika-Intoleranz von einer gewissen Veranlagung auszugehen, die zudem von äußeren Faktoren verstärkt werden kann. Die Analgetika-Intoleranz geht einher mit einer veränderten Zellstoffwechsels ungesättigter Fettsäuren. Durch verschiedene, derzeit nicht bekannte Einflüsse zeigen sich typische Beschwerden meist im Erwachsenenalter. Nicht selten gibt es bereits im Kindesalter Hinweise auf Erkrankungen, etwa als Nasenschleimhautentzündung ohne Infekt und Allergie. Analgetika kommen in den erwähnten Schmerzmitteln vor, aber auch in verschiedenen Produkten, die Salicylsäure-Derivate enthalten, wie z.B. in getrockneten Früchten, Gewürzen oder als Konservierungsmittel in Körperpflegeprodukten oder Stoffen.





Diagnostik der Analgetika-Intoleranz

Eine aussagefähige Diagnose einer Analgetika-Intoleranz kann somit von weitreichender Bedeutung für die Gesundheit Ihres Patienten sein. Neben der Anamnese stand bisher nur der Provokationstest zur Verfügung. Dessen Durchführung ist sehr zeitaufwändig, bedarf erheblicher apparativer Voraussetzungen und sollte nur von ausgewiesenen Spezialisten vorgenommen werden. Zudem ist er mit erheblichen Risiken für Ihren Patienten verbunden. Darüber hinaus erkennt der Provokationstest vorzugsweise Atemwegs-bedingte Analgetika-Intoleranzen. Mit üblichen Laboruntersuchungen ist die Analgetika-Intoleranz nicht erkennbar. Das TalkingCells-Untersuchungsprinzip mit dem LiPiDoC-AIT ist hier eine sinnvolle Alternative für eine Anagetika-Intoleranz-Typisierung (AIT). Dessen Vorteile sind:

risikofrei, da in vitro (d.h. im Reagenzglas)

ambulant, d.h. kein stationärer Krankenhausaufenthalt

ersetzt i.d.R. Provokation mit Analgetika

erkennt frühzeitig Neigung zu Erkrankungen, die durch Analgetika-Intoleranz bedingt sein

können (Risikobewertung)

liefert ein objektives Ergebnis

gibt Hinweis auf Therapiealtenativen

misst den Therapieerfolg

Nachfolgend wurde eine kleine Auswahl von Veröffentlichungen zusammengestellt, in denen die Beteiligung von Eicosanoiden an pathologischen Prozessen zum Ausdruck kommt.





Inhaltsübersicht

Abstracts Eicosanoide und Atemwege 5 Eicosanoide und Haut 22 Eicosanoide und Magen-Darm-Kanal 23 Eicosanoide und Nervensystem 24 Eicosanoide und Zahnheilkunde 26 Eicosanoide und Tumortherapie 28 Eicosanoide und Regulation ihrer Synthese 30

Abstracts � Kurzübersicht 32-34

Publikation Dynamics of eicosanoids in peripheral blood cells during bronchial provocation in

aspirin-intolerant asthmatics. Schäfer D, Schmid M, Göde UC, Baenkler HW.

Eur Respir J. 1999 Mar; 13(3): 638-646.





ABSTRACTS

Eicosanoide und Atemwege

ASA-induced release of histamine from nasal mucous membranes

in analgesic intolerance and polyposis nasi.

1Hosemann WG, 2Baenkler HW, 2Gunther F.

Rhinology. 1990 Dec; 28(4): 231-238.

1Department of O.R.L., 2Medical Clinic III, Dept. of Allergology, University of Erlangen-Nürnberg, Fed. Rep. of Germany.

Tissue samples from the polypous mucous membrane and the inferior nasal

concha were taken from 13 patients with polyposis nasi and from 12 other

patients with an additional intolerance to analgesics. The tissue of the inferior

nasal concha from patients without polyposis nasi served as a control. The

relative histamine content of the samples (in ng/mg dry weight) and the relative

histamine release (in %) after addition of acetylsalicylic acid (ASA) were

determined.

A significantly higher relative histamine content in the tissue samples of polyp

patients without an intolerance to analgesics was seen in comparison to the other

two groups. The relative histamine release of both patient groups with nasal

polyposis was comparable. The control group exhibited both an increased

spontaneous release of histamine as well as a higher relative histamine release

from the tissue of the inferior nasal concha.





Eicosanoids from biopsy of normal and polypous nasal mucosa.

1Baenkler HW, 1Schäfer D, 2Hosemann W.

Rhinology. 1996 Sep; 34(3): 166-170.

1Medical Clinic III, Dept. of Allergology, 2Department of O.R.L., University of Erlangen-Nürnberg, Germany.

In order to clarify the influence of inflammatory mediators of the arachidonic acid

cascade in the mechanism of nasal polyp growth, peptido-leukotriene (pLT),

prostaglandin E2 (PGE2) and thromboxane B2 (TXB2) synthesis was

investigated. In addition to several stimuli, functionally intact human biopsy

specimens of polypous and normal tissue were incubated.

Especially remarkable was the significantly increased release of pLT by polypous

tissue upon arachidonic acid stimulation, in contrast to only slightly elevated

PGE2 release compared to normal tissue. Basic release of pLT and PGE2 was

similar for polypous and normal tissue. Examining TXB2 release, no significant

difference was observed with regard to the origin of tissues.

These data support an altered pattern of the lipoxygenase and cyclo-oxygenase

pathways when tissue becomes irritated and suggest their involvement in the

aetiopathogenesis of nasal polyps.





Dynamics of eicosanoids in peripheral blood cells during bronchial

provocation in aspirin-intolerant asthmatics.

1Schäfer D, 2Schmid M, 2Göde UC, 1Baenkler HW.

Eur Respir J. 1999 Mar; 13(3): 638-646.

2Medical Clinic III, Dept of Allergology, 2Dept. of O.R.L.University of Erlangen-Nürnberg, Germany.

The underlying mechanisms of bronchoconstriction in aspirin-intolerant

asthmatics (AIAs) are still unknown, but the hypothesis of an altered metabolism

of arachidonic acid is generally accepted. So far, no in vitro test for aspirin

intolerance is available. The hypothesis that the profile of eicosanoid mediators is

changed in AIA-even before aspirin challenge was tested.

The release of prostaglandin E2 (PGE2), peptidoleukotrienes and histamine was

measured using competitive enzyme immunoassays in 10 asthmatics with a

history of aspirin intolerance, 10 controls and eight aspirin-tolerant asthmatics

(ATAs) before and after bronchial provocation with lysine-aspirin.

Comparing basal release of eicosanoids before challenge, peptidoleukotrienes

were significantly elevated and PGE2 was vastly reduced in AIAs, whereas ATAs

had elevated basal peptidoleukotrienes but only slightly reduced basal PGE2. The

decrease in forced expiratory volume in one second (FEV1) was not associated

with changes in histamine release. After aspirin challenge, there was a massive

increase of already elevated peptidoleukotrienes in AIAs, but not in ATAs.

Arachidonic acid-induced PGE2 release in AIAs was not significantly changed,

whereas it was significantly reduced in ATAs and healthy controls. Histamine

release was unaffected by aspirin challenge in all three groups.

There is a typically altered profile of eicosanoids in aspirin-intolerant asthmatics

which could make in vitro diagnosis of aspirin intolerance possible.





Arachidonic acid metabolism in nasal tissue and peripheral blood

cells in aspirin intolerant asthmatics.

1Schmid M, 1Göde U, 2Schäfer D, 1Wigand ME.

Acta Otolaryngol. 1999 Mar; 119(2): 277-280.

1Dept. of Otorhinolaryngology, 2Medical Clinic III, Dept of Allergology, University of Erlangen-Nürnberg, Erlangen, Germany.

Aspirin intolerance (AI) is characterized by polypous rhinosinusitis, bronchial

asthma and adverse reactions to aspirin. The common intolerance to all

cyclooxygenase inhibitors allows us to focus study of the pathogenesis of AI on

the metabolism of arachidonic acid (AA).

We studied the metabolism of AA in nine aspirin intolerant asthmatics (AIA) and

eight healthy volunteers (controls) by measuring prostaglandin E2 (PGE2) and

peptido-leukotrienes (pLT = LTC4/D4/E4) in nasal tissue and peripheral blood

cells (PBCs) using a specific immunoassay.

In all patients with AI the tests were performed before and after bronchial

provocation with lysine-ASA. In the control group the tests were done before and

after 500 mg ASA p.o. The release of pLT in nasal polyps of AIA was found to be

significantly higher than in normal mucosa of AIAs and controls. In every tissue a

significant increase of pLT after aspirin challenge was observed. Nasal polyps of

AIA show a significantly lower release of PGE2 than normal mucosa of AIAs and

controls. Peripheral blood cells of AIA show a significantly higher release of pLT

and a significantly lower release of PGE2 than PBCs of controls.

Therefore clinical manifestations of AI may be based on an alteration of AA

metabolism in AIA.





[Significance of eosinophilic granulocytes in relation to allergy and

aspirin intolerance in patients with sinusitis polyposa]

[Article in German]

Kaldenbach T, Schäfer D, Gosepath J, Bittinger F, Klimek L, Mann WJ. Laryngorhinootologie. 1999 Aug; 78(8): 429-434.

Hals-, Nasen-, Ohrenklinik und Poliklinik der Johannes Gutenberg-Universitat Mainz.

BACKGROUND: The development of nasal polyps might be influenced by

different factors such as mucosal inflammation. Infiltration with eosinophils is a

common finding, although in largely different quantitis.

PATIENTS AND METHODS: We investigated 58 patients suffering from nasal

polyps who were assigned for endonasal sinus surgery based on endoscopic and

CT findings. Out of these patients, 52% have already had sinus surgery and had

recurrent polyps. All patients were subjected to both, allergy testing and a

functional in vitro test for aspirin intolerance. During surgery, tissue samples

were gained and send for histological examination with special respect to

eosinophil infiltration.

RESULTS: Eosinophil infiltration was observed in 66% of all patients. 37

patients (64%) exhibited aspirin intolerance only, four (7%) had a positive

allergy test only, in 9 patients (15%) both, allergy and aspirin intolerance were

diagnosed. Allergy and aspirin intolerance were accompanied by eosinophil tissue

infiltration. Eosinophil infiltration was more common and more severe if allergy

and aspirin intolerance were found together compared with the tissue

eosinophilia in patients with either one entity.

CONCLUSION: This study confirms the importance of eosinophil infiltration in

the pathogenesis of severe or recurrent nasal polyps based on allergy and aspirin

intolerance. A coincidance of allergy and aspirin intolerance was found in an

unexpected high number of patients.





Aspirin intolerance in patients with chronic sinusitis.

1Gosepath J, 1Hoffmann F, 1Schäfer D, 2Amedee RG, 1Mann WJ.

ORL J Otorhinolaryngol Relat Spec. 1999 May-Jun; 61(3): 146-150.

1Department of Otolaryngology, University of Mainz, School of Medicine, Mainz, Germany; 2Tulane University Medical Center, New Orleans, School of Medicine, New Orleans, La., USA Department of Otolaryngology

Aspirin intolerance in patients with chronic sinusitis is often a cause of early

recurrence of symptoms after surgical treatment.

This study assesses 84 patients who were tested for acetylsalicylic acid

intolerance after presenting with symptoms like chronic rhinosinusitis, sometimes

bronchial asthma, coexisting allergies or a history of aspirin sensitivity.

Nasal polyposis was found in a majority of cases, often recurrent after previous

surgery. The levels of eicosanoids such as peptido-leukotrienes and prostaglandin

E2 were analyzed in isolated blood cells and compared with a healthy control

group. Aspirin-intolerant patients showed elevated basal levels of peptido-

leukotrienes and reduced basal levels of prostaglandin E2. Test results were

graded in a system ranging from positive (68%), signifying aspirin intolerance, to

borderline (18%) and negative results (14%).

After screening patients with clinical findings indicating a possible aspirin

intolerance, the results of this investigation reveal a strong correlation between

the clinical symptomatology and the in vitro parameters of eicosanoid levels in

isolated blood cells, indicating the need to induce aspirin tolerance to reduce the

risk of recurrent rhinosinusitis.





Prostaglandin E2 activates the ciliary beat frequency of cultured

human nasal mucosa via the second messenger cyclic adenosine

monophosphate.

Haxel BR, Schäfer D, Klimek L, Mann WJ.

Eur Arch Otorhinolaryngol. 2001; 258: 230-235.

Department of Otorhinolaryngology Johannes-Gutenberg University Mainz, Germany.

Prostaglandins influence the ciliary beat frequency (CBF) of ciliated nasal

epithelial cells and a stimulatory effect has been described for prostaglandin E2

(PGE2). Until now, it is not known whether PGE2 has direct ciliostimulatory

properties or acts through a second messenger.

In this study we investigated whether cyclic adenosine monophosphate (cAMP) is

implicated in the signal transduction pathway of PGE2-induced activation of CBF.

Ciliated cells of the nasal mucosa were cultured for up to 5 days whereafter the

culture medium was removed and the cells were incubated with different

concentrations of test solutions.

The ciliated cells were recorded under a phase-contrast microscope and viewed

in slow motion to count the frequency. PGE2 led to a dose-dependent increase in

CBF. This became significant at concentrations of 10(-10) and 10(-5) M (P <

0.01) but not at 10(-13) M (P > 0.05). Addition of cAMP (10(-10) to 10(-5) M)

caused a significant (P < 0.01) increase in CBF, whereas depletion of

endogenous cAMP after pre-incubation with the adenylate cyclase activator

forskolin (10(-5) M) prevented the PGE2-induced increase in CBF (P > 0.05). The

ciliostimulatory effect of PGE2 depends on an intact functioning of adenylate

cyclase.

These results point out that cAMP is directly implicated in the signal transduction

pathway of PGE2-induced stimulation of CBF in cultured human ciliated cells of

the nasal mucosa.





Individual monitoring of aspirin desensitization.

Gosepath J, Schaefer D, Amedee RG, Mann WJ.

Arch Otolaryngol Head Neck Surg. 2001 Mar; 127(3): 316-321.

Department of Otolaryngology-Head and Neck Surgery, Mainz Medical School, Langenbeckstrasse 1, 55101 Mainz, Germany.

BACKGROUND: Patients with aspirin-sensitive rhinosinusitis, which is frequently associated with intrinsic bronchial asthma, can be desensitized by long-term treatment with oral aspirin. The exact mechanisms of this desensitization remain obscure, but modulations of the eicosanoid pathway occur and can be monitored with the help of a practicable in vitro assay on mixed leukocyte cultures. OBJECTIVE: To monitor the effect of low-dose aspirin desensitization therapy, 100 mg/d, objectively by an in vitro assay. DESIGN: In a prospective study, 30 patients with aspirin intolerance, who were treated following a desensitization protocol with a dose of oral aspirin of only 100 mg/d were followed up for 1 year and reassessed every 3 months clinically and in vitro. RESULTS: Twenty-five patients showed a normalization of in vitro eicosanoid levels during this period, 4 showed some improvement, and 1 showed no therapeutic effect on eicosanoid release. Clinical follow-up revealed a low recurrence rate of nasal polyposis, with recurrent disease only in 4 individuals who also showed no normalization of eicosanoid release levels. Furthermore, a reduction of the average incidence of purulent episodes of sinusitis was seen after 1 year. Of 12 patients with asthma, 9 experienced marked improvement in pulmonary function. Of 16 individuals with a marked impairment of nasal breathing, 14 felt an increase of nasal patency, and 7 of 11 patients with pretreatment hyposmia had an improved sense of smell after 1 year. CONCLUSIONS: Desensitization therapy in patients with aspirin-sensitive rhinosinusitis can be successfully performed with low oral doses of aspirin, and the individual course throughout the desensitization can be monitored with the help of an in vitro analysis of eicosanoid release from mixed leukocyte cultures.





Analgetika-Intoleranz: Langzeitergebnisse bis zu 3 Jahren bei

adaptiver Desaktivierung mit einer täglichen Erhaltungsdosis von

100 mg Aspirin

Gosepath, D. Schäfer, W. J. Mann Laryngorhinootologie. 2002; 81: 732-738.

Universitäts-HNO-Klinik Mainz (Direktor: Prof. Dr. med. Dr. h. c. W. J. Mann)

Hintergrund: Obwohl die pathophysiologischen Grundlagen des Krankheitsbildes des Analgetika-Intoleranzsyndromes bis heute nicht vollständig erforscht sind, deuten eine Vielzahl von Untersuchungen auf eine Störung des Eicosanoidstoffwechsels als wesentlichen ursächlichen Faktor hin. Die Analyse der Eicosanoidfreisetzung in vitro ist in den letzten Jahren als hilfreiches Instrument in der Diagnostik dieses Krankheitsbildes etabliert worden. Diese ermöglicht zusätzlich eine objektive Therapiekontrolle während der adaptiven Desaktivierungstherapie. Methodik: Im Rahmen der vorliegenden prospektiven Untersuchung wurden 30 Patienten, bei denen zwischen 1996 und 2000 an unserer Klinik eine Analgetika-Intoleranz diagnostiziert wurde, zwischen einem und drei Jahre lang einer adaptiven Desaktivierungstherapie unterzogen. Hierbei wurde eine Erhaltungsdosis von nur 100 mg Aspirin pro Tag verabreicht. Es erfolgte eine regelmäßige Kontrolle der klinischen wie auch der in vitro Parameter. Es traten bei dieser Dosierung keine durch Aspirin verursachten unerwünschten Nebenwirkungen auf. Ergebnisse: Es zeigte sich eine klare positive Korrelation zwischen klinischer Symptomatik und Veränderungen des Eicosanoidstoffwechsels. Gemessen an der Rezidivrate der Polyposis nasi, dem Schweregrad des intrinsischen Asthma bronchiale und dem Riechvermögen wurde die Therapie bei 25 Patienten langfristig als erfolgreich bewertet. Eine Unterbrechung oder ein Absetzen der Therapie führte zu einer erneuten klinischen Verschlechterung. Schlussfolgerungen: Die Ergebnisse dieser Untersuchung belegen die Wirksamkeit einer Desaktivierungstherapie bei Patienten mit Aspirinintoleranz mit einer nebenwirkungsfreien Dosierung von 100 mg. Der langfristige Erfolg einer solchen Therapie kann nur durch eine anhaltende, möglichst lebenslange Einnahme geringer Dosen von Aspirin gesichert werden.





[Aspirin Sensitivity: Long Term Follow-up After Up to 3 Years of

Adaptive Desensitization Using a Maintenance Dose of 100 mg of

Aspirin a Day]

Gosepath, D. Schäfer, W. J. Mann

Laryngorhinootologie. 2002; 81: 732-738.

Background: The full clinical picture of aspirin intolerance, Sampter's triad, is

associated with nasal polyposis, clinical sensitivity to most non steroidal

antiinflammatory drugs (NSAID) and intrinsic bronchial asthma. But the triad can

be incomplete and nasal polyposis can be the first clinical symptom of aspirin

sensitivity. Although the exact mechanisms of aspirin intolerance as well as those

of desensitization remain obscure, an in vitro assay on eicosanoid metabolism

has been proven to be helpful in diagnosis and treatment as it correlates well to

the individual severity of clinical symptoms.

Methods: For this investigation 30 patients, who were undergoing adaptive

desensitization for aspirin intolerance, were followed-up between 1 and 3 years.

They received a maintenance dose of oral aspirin of only 100 mg a day after an

initial application of higher doses. Their clinical course as well as their in vitro

parameters of eicosanoid release were monitored throughout the individual

observation period.

Results: Desensitization was successful in 25 of the 30 patients regarding the

recurrence rate of nasal polyps, severity of bronchial asthma and sense of smell.

There was a clear positive correlation between clinical and in vitro parameters.

Discontinuing of aspirin therapy lead to worsening of clinical symptoms,

regardless of the prior duration of treatment.

Conclusions: This article reviews the role of the in vitro assay and presents a

desensitization protocol that can be maintained as a long term treatment without

adverse side effects. Results suggest that the recurrence rate of nasal polyps

after surgical therapy can be reduced using this protocol, however, only long

term treatment can secure a beneficial outcome over time.





Diagnostik des Analgetika-Intoleranz-Syndroms mittels

funktioneller Zelltestung (Analgetika-Intoleranz-Test: AIT)

1Hecksteden K, D. 1Schäfer D, B.A. 1Stuck1, 1,2Klimek L, 1Hörmann K.

Allergologie. 2003 Jul; 26(7): 263-271.

1HNO-Klinik, Universitätsklinikum Mannheim, 2Zentrum für Rhinologie und Allergologie, Wiesbaden

Das klinisch manifeste Analgetika-Intoleranz-Syndrom (AIS) ist typischerweise

durch rezidivierende Polyposis nasi, Asthma bronchiale und Analgetika-

Unverträglichkeit gekennzeichnet. Es beruht nach gängiger Meinung auf

Dysbalancen im Arachidonsäure-Metabolismus, ausgelöst durch die Einnahme

von non-steroidalen Antirheumatika (NSAR). Bisher sind in der Diagnostik des

AIS Anamnese und Provokationstests etabliert. Die Methoden der nasalen,

bronchialen und oralen Provokationstests sind für die Praxis umfangreich und für

die Patienten meist unangenehm und risikobehaftet. Ziel dieser Arbeit war es

herauszufinden, inwieweit ein komplikationsloses In-vitro-Verfahren mit

funktioneller Zelltestung (Analgetika-Intoleranz-Test: AIT) zur Diagnosesicherung

herangezogen werden kann.

Hierfür wurde die herkömmliche Diagnostik mit den Ergebnissen des AIT an 50

Patienten mit Verdacht auf Analgetikaintoleranz verglichen. Zur Kontrolle dienten

Tests an 38 Patienten mit leerer Anamnese und ohne klinische Zeichen einer

Polyposis oder eines Asthma bronchiale. Für die Sensitivität des Tests wurden

100%, für die Spezifität 73,3% berechnet, der negative Vorhersagewert des AIT

lag bei 100% und der positive Vorhersagewert bei 71%. Dies erklärt sich durch

eine Gruppe von Patienten, bei denen die herkömmliche Diagnostik zu einem

negativen, der AIT hingegen zu einem positiven Ergebnis gekommen war.

Eine Follow-up-Studie muß klären, ob bei diesen Patienten ein klinisch nicht

manifestes AIS vorliegt, oder ob der Test falsch-positive Ergebnisse liefert.

Zusammenfassend ist der AIT bisher � trotz der guten Ergebnisse � nicht zur

Diagnosesicherung eines AIS ausreichend, betrachtet man aber seine Vorteile in

Praktikabilität und Risikominimierung, so ist eine weitere Validierung des Tests

zu fordern.





Untreated chronic rhinosinusitis: a comparison of symptoms and

mediator profiles.

Kuehnemund M, Ismail C, Brieger J, Schaefer D, Mann WJ.

Laryngoscope. 2004 Mar; 114(3): 561-565.

Department of Otorhinolaryngology-Head and Neck Surgery, Mainz Medical School, Mainz, Germany.

OBJECTIVES/HYPOTHESIS: Since the early 1990s, the knowledge of inflammatory mediators involved in chronic rhinosinusitis has increased extensively. However, until the present no data on trends in mediator levels in the natural course of the exacerbation-free interval of chronic rhinosinusitis have been published. The purpose of the study was to examine how levels of inflammatory mediators and clinical findings in chronic rhinosinusitis relate over time in the absence of acute exacerbation. STUDY DESIGN: Prospective study. METHODS: The authors investigated 12 untreated patients with chronic rhinosinusitis and repeated clinical examinations and measurements of inflammatory mediators in the nasal mucosa (messenger RNA [mRNA]of interleukin-1beta; interleukins 3, 5, 6, and 8 [IL-3, IL-5, IL-6, and IL-8]; interferon gamma; tumor necrosis factor-alpha [TNF-alpha]; monocyte chemotactic proteins 1, 3, and 4 [MCP-1, MCP-3, and MCP-4]; and granulocyte-macrophage colony-stimulating factor [GM-CSF]), as well as measurements of peptido-leukotriene [PLT] and prostaglandin E2 [PGE2]) in nasal secretion over a period of 4 weeks. RESULTS: Clinically speaking, the symptom score significantly improved over the period of 4 weeks, whereas other clinical parameters (computed tomography score, endoscopy score, rhinomanometric values, saccharine transport time, ciliary beat frequency, and olfaction) varied insignificantly. Regarding proinflammatory and inflammatory mediators, only mRNA of IL-1beta, IL-6, IL-8, MCP-1, and TNF-alpha were detected in relevant amounts in nasal mucosa, and their levels decreased only insignificantly over the 4-week period. In nasal secretion, a slight decrease of PGE2 and PLT levels was observed over the same time period. CONCLUSION: Subjective symptoms may show a spontaneous improvement of approximately 25% in patients with so-called "stable" chronic rhinosinusitis over a 4-week period, whereas objective clinical parameters vary insignificantly. Messenger RNA of IL-1beta, IL-6, IL-8, MCP-1, and TNF-alpha, as well as PLT and PGE2 levels, are detectable and appear to play a role in the persistence of inflammation in chronic rhinosinusitis. Their levels decrease only insignificantly over time, even in the absence of acute exacerbation of disease, possibly rendering the mucosa more prone to recurrent acute episodes.





Experimentally induced nasal irritation.

1Mohammadian P, 2Schaefer D, 1Hummel T, 1Kobal G.

Rhinology. 1999 Dec; 37(4): 175-178.

1Dept. of Experimental and Clinical Pharmacology and Toxicology, 2Medical Clinic III, Dept. of Allergology, University of Erlangen-Nürnberg, Germany.

The aim of this study was to develop a method that is suited for the induction of

nasal irritation. For this purpose inflammatory responses were analysed after

challenging the nasal mucosa with experimentally induced cold, dry air (8 l/min,

22 degrees C, 20 %RH). To assess inflammatory effects we determined

inflammatory mediators (prostaglandin E2 [PGE2], thromboxane B2 TXB2[,

peptide leukotrienes, pLT [LTC4, LTD4, LTE4]) in nasal lavage fluid which was

sampled before, immediately after suprathreshold stimulation, and one hour

after termination of the stimulation. In addition, subjects estimated the intensity

of pain during the stimulation.

Cold, dry air produced strong painful sensations which increased throughout the

stimulation period. A significant increase of the inflammatory mediator pLT was

observed after stimulation; mean concentrations of PGE2 and TXB2 also showed

a tendency to increase. One hour after termination of the stimulation the

concentration of these inflammatory mediators returned to baseline which

indicates the reversibility of the effects of nasal irritation.

These data suggest, that this model may be a useful tool in investigations of

mucosal irritation as, for example, induced by environmental agents.





Leukotrienrezeptorantagonisten zur Rezidivprophylaxe bei der

ASS-assoziierten Polyposis - Erste klinische Ergebnisse zur

Wirkung auf entzündliche Gewebeprozesse

T. Grundmann, M. Töpfner

Laryngorhinootologie 2001; 80: 576-582.

Universitäts-Hals-Nasen-Ohrenklinik Hamburg-Eppendorf (Direktor: Prof. Dr. U. Koch)

Hintergrund: Die mit einer Aspirinintoleranz (AI) assoziierte Polyposis nasi et sinuum stellt aufgrund häufiger Rezidive ein besonderes therapeutisches Problem dar. Basierend auf neueren Untersuchungen, die ätiologisch Störungen im Arachidonsäuremetabolismus mit erhöhten Leukotrienspiegeln nachweisen konnten, wurden kausal in diesen Pathomechanismus eingreifende Therapeutika wie z. B. die Leukotrienrezeptorantagonisten (LTA) entwickelt. In der Behandlung des ASS-assoziierten Asthma bronchiale konnten erste klinische Erfolge in Bezug auf die pulmonale Symptomatik nachgewiesen werden, Untersuchungen zur Wirksamkeit auf die ASS-assoziierte Polyposis nasi et sinuum (ASSAP) liegen jedoch noch nicht vor. Ziel unserer Untersuchung war es daher, Leukotrienrezeptorantagonisten auf ihre therapeutische Wirksamkeit in Hinblick auf die entzündlichen Veränderungen der ASS-assoziierten Polyposis zu untersuchen. Methode: Hierzu wurden 18 Patienten unseres rhinologischen Klientels mit nachgewiesener AI und zusätzlich bestehenden pulmonalen Beschwerden, bei denen aufgrund einer Rezidivpolyposis eine operative Nasennebenhöhlensanierung indiziert war, postoperativ über ein Jahr mit dem LTA Montelukast therapiert. Als klinische Verlaufsparameter dienten subjektive Befundskalierungen, endoskopische Befundschemata und serologische Befunde (ECP). Zur zusätzlichen Beurteilung möglicher antiinflammatorischer Gewebeeffekte wurden immunhistologisch EG1/EG2-positive Zellen und die Interleukin 5-Spiegel im Gewebe der unteren Nasenmuscheln bestimmt. Ergebnisse: Bei den klinischen Untersuchungen war über einen Beobachtungszeitraum von zwölf Monaten unter LTA-Therapie endoskopisch keine Rezidivpolyposis nachweisbar, symptomatisch beschrieben die Patienten postoperativ einen dauerhaften Rückgang der rhinitischen und pulmonalen Beschwerden im Vergleich zur Kontrollgruppe die nur mit topischen Steroiden behandelt wurde. Ein reduzierter Anteil EG2-positiver Zellen sowie erniedrigte Interleukin 5-Spiegel lassen einen antiinflammatorischen Effekt der LTA bei der ASSAP vermuten. Schlussfolgerungen: Leukotrienantagonisten können bei Patienten mit ASS-assoziierter Polyposis eine therapeutische Alternative zur ASS-Toleranzinduktion zur postoperativen Rezidivprophylaxe darstellen.





[Treatment of ASS-Associated Polyposis (ASSAP) with a Cysteinyl

Leukotriene Receptor Antagonist - A Prospective Drug Study on its

Antiinflammatory Effects]

T. Grundmann, M. Töpfner

Laryngorhinootologie 2001; 80: 576-582.

Background: In a high rate of cases with recurrent polyposis an association with ASS-intolerance is detectable despite missing pulmonary symptoms. New examinations of a disturbed arachidonic acid metabolism lead to the development of new therapeutical options. Treatment with leukotriene-receptor antogonists (LTA) showed primarily good results in therapy of ASS-associated asthma. Methods: 18 patients with ASS-intolerance trias - diagnosed by oral provocation - were treated with the LTA Montelukast, after undergoing sinus surgery. Patients underwent a diagnostic pathway of provocation including four groups: recurrent chronic sinusitis, excessive polyposis, polyposis associated with asthma and anaphylactic symptoms after oral ASS-intake. Clinically we examined the following parameters periodically after sinus surgery: nasal and pulmonal symptoms by scoring levels, recurrency of polypoid hyperplasia by endoscopic follow-ups and serum ECP-levels. To evaluate antiinflammatory tissue effects of LTA EG1/EG2 labelled cells and cytokine levels of Interleukin 5 in mucosa samples of the lower turbinate were analysed under LTA-therapy. Results: Under therapy with LTA we saw a beneficial effect on nasal and pulmonary symptoms and a significant reduction of recurrent polyposis in endoscopic examinations in relation to the untreated group. Results were proven by a permanent reduction of serum ECP-level. A reduction of the rate of EG2-positive cells according to decreased Interleukin 5 levels in the nasal mucosa unter LTA-treatment assumed antiinflammatory effects on ASS-associated polyposis. Conclusions: We could demonstrate antiinflammatory effects of Leukotriene-Receptor-Antagonists primarily during postoperative treatment of patients with ASS-associated nasal polyps.





Analgetikasensitive Atemwegserkrankung: Diagnostik,

Risikoanalyse und Therapieplanung mittels FET

G Eger, D Schäfer, HW Baenkler Pneumologie 2004; 58 DOI: 10.1055/s-2004-819570 Medizinische Klinik III, Abteilung Allergologie, Universität Erlangen-Nürnberg,

Die Intoleranz gegenüber Salizylatanaloga kann sich mit chronisch entzündlicher

Reaktion an Haut, Atmungs- und Verdauungsorganen manifestieren und als

akutes Analgetikaasthma oder anaphylaktoide Akutreaktion in Erscheinung

treten. Als nahezu ubiquitäre Nahrungsmittelinhaltsstoffe sind Salizylate jedoch

nicht zu vermeiden. Zusätzliche Bedeutung erhält diese Intoleranzreaktion durch

die Tatsache, dass zu den wirksamen Analoga eine Vielzahl von Schmerzmitteln

und Entzündungshemmern aus der Gruppe der NSAID zählen.

Als Sonderform des NSAID-sensitiven Komplexes ist die Samter-Trias mit der

Kombination von Analgetikaintoleranz, Asthma und nasaler Polyposis bekannt.

Entzündungsaktivität und Akutreaktivität lassen sich in diesem Fall durch

Dauermedikation mit Aspirin verbessern (ASS-Desaktivierungstherapie).

Zur Sicherung der Salizylat- bzw. Analgetikaintoleranz ist bislang eine

risikoreiche ASS-Expositionstestung nicht zu umgehen, insbesondere wenn eine

langjährige Desaktivierungsbehandlung ansteht.

Auf Basis der Pathogenese in einem veränderten Arachidonsäuremetabolismus

wurde ein funktioneller Eicosanoidtest (FET) entwickelt, der die biochemisch

relevanten Vorgänge bei Salizylattoleranz und -intoleranz in vitro abbildet,

unabhängig von der jeweiligen Manifestationsform der Erkrankung. Die

biochemischen Daten des FET wurden mit den Ergebnissen von ASS-

Expositionstestungen und mit Therapieverläufen korreliert.





Effect of prostaglandin E2 on eicosanoid release by human

bronchial biopsy specimens from normal and inflamed mucosa.

1Schäfer D, 1Lindenthal U, 2Wagner M, 2Bolcskei PL, 1Baenkler HW.

Thorax. 1996 Sep;51(9):919-23.

1Dept. of Allergology, University of Erlangen-Nürnberg, Germany, 2Dept. of Pneumologie, University Hospital-Nürnberg.

BACKGROUND: Eicosanoids such as prostaglandin E2 (PGE2), thromboxane A2

(TXA2), and peptidoleukotrienes (pLT) are known to be biologically highly active

lipid mediators, especially in human lung epithelium. PGE2 is thought to have

mostly bronchoprotective effects, whereas pLT and TXA2 are bronchoconstrictive.

This study was undertaken to assess the release and interaction of eicosanoids in

human bronchial biopsy specimens of normal and inflamed mucosa.

METHODS: Bronchial biopsy specimens were obtained from 16 patients, seven

controls without signs of inflammation and nine patients with severe

inflammatory processes in the epithelium. The release of pLT, TXA2 (measured

as TXB2), and PGE2 was investigated using a "functional in vitro test" and the

addition of several stimuli.

RESULTS: Specimens incubated with arachidonic acid released higher amounts

of pLT, TXB2, and PGE2 than unstimulated specimens. Preincubation with PGE2

revealed significant inhibition of arachidonic acid-induced release of pLT and

TXB2 (> 50%). The inhibitory effect was higher in normal than in inflamed

epithelium.

CONCLUSIONS: Exogenous PGE2 has inhibitory effects on the release of pLT

and TXB2 in human bronchial biopsy specimens. This finding could explain the

bronchoprotective effect of inhaled PGE2 in normal subjects and asthmatic

subjects as direct eicosanoid interactions. It also supports the concept of PGE2 as

a bronchoprotective endogenous substance. The complex effects of PGE2 as a

modulating mediator in inflammation may be worth investigating.





Eicosanoide und Haut

Stimulated prostaglandin E2 release from rat skin, in vitro

1Sauer SK, 2Schafer D, 1Kress M, 1Reeh PW.

Life Sci. 1998; 62(22): 2045-255.

1Institut für Physiologie und Experimentelle Pathophysiologie, 2Medical Clinic III, Dept. of Allergology, Universität Erlangen-Nürnberg, Erlangen, Germany.

The excitatory effect of bradykinin (BK) and of low pH on nociceptors appears to

partly depend on secondary release of prostaglandins from the surrounding

tissue. Rat skin, in vitro, is introduced as a novel model to measure basal and

stimulated release of PGE2 and, in future, other substances relevant to

nociception, such as neuropeptides.

Flaps of hairy skin (n=57) from the rat saphenous region of the hindpaw were

subcutaneously excised and fixed on acrylic rods, the corium side exposed. The

preparations were equilibrated in carbogen gassed "synthetic interstitial fluid"

(SIF) for 30 minutes. The skin flaps were then immersed for 5 minutes each in 9

consecutive glass tubes, which were mounted in a shaking bath at 32 degrees C.

Each tube was filled with 5 ml of gassed SIF, the third tube contained

inflammatory mediator(s) dissolved in SIF or solutions of low pH. After passage

of the skin flap, the eluates were deep frozen (-70 degrees C) and the PGE2

content measured, off-line, using an enzyme immuno-assay.

As stimulants, BK at 10(-5) M (n=9) and 10(-6) M (n=4) and BK in equimolar

combination with histamine (HA) and serotonin (5-HT; 10(-5) M: n=8, 10(-6) M:

n=6, 10(-7) M: n=6) dose-dependently increased PGE2 release. Considering the

total amount of PGE2 secreted the combination of inflammatory mediators

caused a significantly greater release of PGE2 at 10(-5) and 10(-6) M (p<0.01,

Kruskal-Wallis test) than BK stimulation alone.

Racemic flurbiprofen caused a profound depression of basal and stimulated

release. Solutions of high proton concentration are known to stimulate and

sensitize nociceptors. However, phosphate buffered SIF at pH 6.1 and 6.4 caused

a substantial and significant decrease of the PGE2 release, probably due to low-

pH block of phospholipases. Thus, algogenic potency of mediators does not

necessarily match their pro-inflammatory action.





Eicosanoide Magen-Darm-Kanal

Abnormal eicosanoid pattern by blood leukocytes in

gastroduodenal ulcer.

Baenkler HW, Zeus J, Schenk J, Schäfer D.

Med Sci Monit. 2004 Oct;10(10):CR557-62. Epub 2004 Sep 23.

University Hospital, Department of Medicine III, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.

BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are

implicated in several diseases showing altered tissue and leukocyte eicosanoid

patterns, such as nasal polyposis and asthma. NSAIDs are also associated with

gastrointestinal lesions, but it is unknown whether there is an altered eicosanoid

pattern.

MATERIAL/METHODS: The ex vivo modulated syntheses of prostaglandin E2

(PGE2) and peptido-leukotrienes (pLT) by leukocytes from 41 patients with

gastroduodenal ulcer were compared with those of 61 healthy controls. Samples

were incubated with diluent, arachidonic acid, or acetylsalicylic acid. The

individual syntheses of PGE2 and pLT were quantified using competitive enzyme-

immuno-assays followed by calculation of individual eicosanoid patterns.

RESULTS: Controls synthesized approximately 4.9-fold whereas patients only

approximately 2.9-fold more PGE2 than pLT due to higher basal synthesis of pLT

(67 and 125 pg/ml, respectively). The baseline PGE2/pLT ratio was slightly

higher in patients (6.1) than in controls (5.7). The arachidonic acid-induced

PGE2/pLT ratio in patients (14.2) was significantly higher than in controls (3.3).

The acetylsalicylic acid-induced PGE2/pLT ratio in patients (3.5) was significantly

lower than in controls (8.3) due to diminished PGE2 and elevated pLT. Integrated

individual PGE2 and pLT values revealed a highly significantly altered eicosanoid

pattern score in approximately 95% patients and approximately 12% controls.

CONCLUSIONS: There is strong evidence of an altered eicosanoid pattern

generated by leukocytes of gastroduodenal ulcer patients, which became obvious

upon in vitro modulation by arachidonic or acetylsalicylic acid. The phenomenon

of an abnormal eicosanoid pattern in gastroduodenal ulcer is yet not fully

understood, but may have implications in pathophysiology and diagnostics.





Eicosanoide und Nervensystem

Up-regulation of cyclooxygenase-1 in neuroblastoma cell lines by

retinoic acid and corticosteroids.

1Schneider N, 1Lanz S, 1Ramer R, 2Schaefer D, 1Goppelt-Struebe M.

J Neurochem. 2001 Apr;77(2):416-24.

1Medizinische Klinik IV, Universitat Erlangen-Nürnberg, Erlangen, Germany 2Kompetenzzentrum Umweltmedizin, Rupprecht-Karls-Universitat Heidelberg, Mannheim, Germany.

Cyclooxygenases-1 and -2 are both expressed in neuronal cells in vivo. In the

neuroblastoma cell lines NG108 and N2a, however, only cyclooxygenase-1 was

detectable. Differentiation of the cells with retinoic acid increased

cyclooxygenase-1 mRNA and protein expression within 24 and 48 h, respectively.

A further increase was observed when the cells were concomitantly treated with

the glucocorticoid dexamethasone (a 2-3-fold increase compared with retinoic

acid alone). In the absence of retinoic acid, dexamethasone only slightly up-

regulated cyclooxygenase-1 expression. The inhibitor of protein synthesis

cycloheximide abrogated the effect of dexamethasone, indicating the

involvement of newly synthesised proteins. Retinoic acid increased the

transcription of cyclooxygenase-1 mRNA, determined with a luciferase-coupled

promoter construct. Dexamethasone only slightly augmented cyclooxygenase-1-

promoter activity but increased cyclooxygenase-1 mRNA stability. Other

corticosteroids, hydrocortisone and aldosterone, also up-regulated

cyclooxygenase-1 whereas neurosteroids or oestrogen were ineffective. Up-

regulation was mediated primarily by the glucocorticoid receptor, because the

receptor antagonist RU486 strongly reduced the effects of all corticosteroids.

This indicated that in NG108 cells, the mineralocorticoid aldosterone may bind to

the glucocorticoid receptor. Treatment of NG108 or N2a cells with corticosteroids

did not alter the morphological phenotype obtained during differentiation.

We thus show that corticosteroids, which down-regulate cyclooxygenase

expression in most cell types, up-regulate cyclooxygenase-1 during neuronal

differentiation.





Thromboxane A2 release in ischemia and reperfusion of

free flaps in rats, studied by microdialysis.

Ionac M, Schaefer D, Geishauser M.

J Reconstr Microsurg. 2001 Feb;17(2):139-43.

Second Surgical Clinic, University of Medicine and Pharmacy, Timisoara, Romania.

Several studies have implicated enhanced eicosanoid production in reperfusion

injury. The reported study investigated the use of microdialysis in the in vivo

measurement of thromboxane levels during reperfusion in ischemic and

reperfused experimental free muscle flaps.

Microdialysis probes were inserted, via a guide, into the gracilis and

semitendinosus free flap in the rat. The probe was perfused at a flow of 5

microl/min, with samples collected at intervals of 20 min, and analyzed by the

ELISA technique. Animals were randomly distributed into three groups. After

ischemic periods of 2, 4, and 6 hr, respectively, the free muscle flaps were

revascularized on the contralateral femoral vessels.

The mean thromboxane level during ischemia was 1785.34 +/- 124.81 pg/ml.

The mean levels of thromboxane rose significantly (p < 0.05), compared to base

level, with 151.65 percent in the 2-hr ischemia group, 192.33 percent in the 4-hr

ischemia group, and 294.69 percent in the 6-hr ischemia group, and correlated

well with histologic observations.

The results suggest that a microdialysis technique, combined with a sensitive

assay for measuring thromboxane, is a useful method for in vivo monitoring of

inflammatory processes during ischemia and reperfusion. The evolution of

thromboxane release following 6 hr of ischemia indicates that this mediator may

be involved in facilitation of cell death, following ischemia and reperfusion, since

its tissue level correlates with histologic tissue damage.





Eicosanoide und Zahnheilkunde

Influence of resin-based restoration materials on cytotoxicity in

gingival fibroblasts.

Willershausen B, Schafer D, Pistorius A, Schulze R, Mann W.

Eur J Med Res. 1999 Apr 27; 4(4): 149-155.

Department of Restorative Dentistry and Periodontology, Augustusplatz 2, D-55131 Mainz, Germany.

Tooth-colored resin-based restoration materials have gained enormous significance in recent

decades. Since the composites presently available still evidence various weaknesses, intensive

research continues concerning them.

In this study we tested the biocompatibility of six resin-based materials (Charisma, Solitaire,

Definite, Tetric ceram, Ariston pHc and Compoglass) by means of cultured human gingival

fibroblasts. Over a period of 9 days the cured resin-based materials were incubated with the

fibroblasts. Cell growth, cell viability, and protein content was measured. The release of

prostaglandin (PGE2) was also determined.

There was a reduction of cell functions, related to the different resin-based materials. Protein

content of the fibroblasts was significantly reduced (Kruskal Wallis test; p = 0.0001),

depending on the material tested. Definite induced only minimal irritation, and there was

correspondingly little contrast to the protein content of the controls. Solitaire induced the

highest levels of irritation and the greatest reduction in protein content. Lactate production of

fibroblasts also showed some significant reduction in contact with particular materials (p =

0.005). There were only small differences among the tested materials as concerns measured

PGE2 release of the fibroblasts.

The data suggest that the methods here employed may be quite useful in testing the cytotoxic

potential of newly developed resin-based materials.





Cytotoxicity of root canal filling materials to three different human cell lines. Willershausen B, Marroquin BB, Schafer D, Schulze R. J Endod. 2000 Dec;26(12):703-7.

Department of Operative Dentistry and Periodontology, Johannes Gutenberg-University, Mainz,

Germany.

The aim of this study was to investigate the biological compatibility of five root canal sealers

(Sealapex, Endion, Super-EBA, Ketac-Endo, and AH Plus) and regular and calcium

hydroxide-based gutta-percha in three different human cell lines.

Cultures without root canal sealers were used as controls. Cell growth, cell morphology, cell

viability, protein content of the cells, and prostaglandin E2 (PGE2) release were used as

parameters to determine the cytotoxicity of the materials.

The protein content of the three cell lines--nasal fibroblasts, gingival fibroblasts, and

epithelial tumor cells was significantly reduced (p < or = 0.001) by all materials tested.

Determinations of PGE2 release showed significant material specific differences. No

significant increased PGE2 release values were found with Sealapex, Endion, and Super-

EBA.

On the contrary significantly increased PGE2 levels were measured with Ketac Endo, AH Plus, regular and calcium hydroxide-based gutta-percha points (p < or = 0.001).





Untersuchungen zur Zytotoxizität von Wurzelkanfüllmaterialien. 1Willershausen B, 1Briseno B, 2Schulze R 3Schäfer D,. Dtsch Zahnärztl Z 2000 ;55:252-256. 1 Poliklinik für Zahnerhaltungskunde, Johannes Gutenberg-University, Mainz, Germany. 2 Poliklinik für zahnärztliche Chirurgie, Johannes Gutenberg-University, Mainz, Germany, 3 Klinik für Hals-Nasen-Ohrenheilkunde, Gutenberg-University, Mainz, Germany

Die biologische Verträglichkeit von 4 Wurzelkanalzementen (Sealapex, Super EBA, Ketac-

Endo, AH Plus; Aushärtungsueit 24 Std.) sowie Guttaperchespitzen mit Claciumhydroxid wurde in drei verschiedenen Zellsystemen (Gingivafibroblasten, nasalen Epithelzellen und

epithelialen Tumorzellen) menschlichen Ursprungs über einen Zeitraum von 6 Tagen überprüft. Als Vergleich dienten Goldlegierungsproben und Zellen ohne Werkstoffe. Beobachtungsparameter waren Zellwachstum, Zellmorphologie, Zellvitalität (Fluoreszenztmarkierung), Proteingehalt der Zellen, und Freisetzung von Prostaglandin E2

(PGE2, clone E3R1).

Die getesteten Wurzelkanfüllungsmaterialien führten in den jeweils getesteten Zellsystemen zu materialabhängigen signifikanten Veränderungen der Zellfunktionen. Die Proteinsynthese war bei allen Materialien sowohl bei den nasalen und gingivalen Fibroblasten als auch bei den

epithelialen Tumorzellen signifikant vermindert (t-test, p<0,001). Die Bestimmung der PGE2-

Werte zeigte, dass gingivale und nasale Fibroblasten gleichfalls signifikante,

werkstoffabhängige veränderungen erkenn ließen, epitheliale Tumorzellen reagierten weniger sensibel. Keine Irritationen fanden sich für Sealpex und Super EBA, während Ketac Endo und

Ah Plus vermehrte Prostaglandinwerte bewirkten.

Die vorliegenden Untersuchungen unterstreichen, dass Fibroblastenkulturen ein geeignetes

Modellsystem darstellen, um die biologische Verträglichkeit von Wurzelkanalfüllmaterialien zu überprüfen.





Eicosanoide und Tumortherapie

The cyclooxygenase system participates in functional mdr1b

overexpression in primary rat hepatocyte cultures.

1Ziemann C, 2Schafer D, 1Rudell G, 1Kahl GF, 1Hirsch-Ernst KI.

Hepatology. 2002 Mar; 35(3): 579-588.

1Department of Toxicology, Institute of Pharmacology and Toxicology, University of Göttingen, Robert-Koch-Strasse 40, D-37075 Gottingen, 2Kompetenzzentrum Umweltmedizin, Rupprecht-Karls-Universitat Heidelberg, Mannheim, Germany.

Overexpression of mdr1-type P-glycoproteins (P-gps) is thought to contribute to primary chemotherapy resistance of untreated hepatocellular carcinoma. However, mechanisms of endogenous multidrug resistance 1 (mdr1) gene activation still remain unclear. Because recent studies have demonstrated overexpression of cyclooxygenase-2 (COX-2) in hepatocytes during early stages of hepatocarcinogenesis, we investigated whether the COX system, which catalyzes the rate-limiting step in prostaglandin synthesis, participates in mdr1 gene regulation. In the present study, primary rat hepatocyte cultures, exhibiting time-dependent mdr1b overexpression, demonstrated basal COX-2 and COX-1 mRNA expression and liberation of prostaglandin E(2) (PGE(2)), indicative of an active COX-dependent arachidonic acid metabolism. PGE(2) accumulation in culture supernatants was further enhanced by arachidonic acid (1mumol/L) and epidermal growth factor (EGF) (16 nmol/L). PGE(2) and prostaglandin F(2alpha) (PGF(2)alpha) (3-6mug/mL), added directly to the culture medium, significantly up-regulated intrinsic mdr1b mRNA overexpression and mdr1-dependent transport activity. Up-regulation was maximal after 3 days of culture. Like prostaglandins, the COX substrate, arachidonic acid, also induced mdr1b gene expression. Apart from this, structurally different COX inhibitors (indomethacin, meloxicam, NS-398) mediated significant inhibition of time-dependent and EGF-induced mdr1b mRNA overexpression, resulting in enhanced intracellular accumulation of the mdr1 substrate, rhodamine 123 (Rho123). Thus, the present data support the conclusion that the release of prostaglandins through activation of the COX system participates in endogenous mdr1b gene regulation. COX-2 inhibition might constitute a new strategy to counteract primary mdr1-dependent chemotherapy resistance.





Impact of the cyclooxygenase system on doxorubicin-induced

functional multidrug resistance 1 overexpression and doxorubicin

sensitivity in acute myeloid leukemic HL-60 cells.

1Puhlmann U, 2Ziemann C, 1Ruedell G, 1Vorwerk H, 3Schaefer D, 1Langebrake C, 1Schuermann P, 1Creutzig U, 1Reinhardt D.

J Pharmacol Exp Ther. 2005 Jan;312(1):346-54. Epub 2004 Oct 22.

1AML-BFM Study, Department of Pediatric Hematology and Oncology, University Children's Hospital Muenster, Albert-Schweitzer-Str. 33, 48129 Münster, 21Department of Toxicology, Institute of Pharmacology and Toxicology, University of Göttingen, Robert-Koch-Strasse 40, D-37075 Göttingen, 3Kompetenzzentrum Umweltmedizin, Rupprecht-Karls-Universitat Heidelberg, Mannheim, Germany.

Multidrug resistance (MDR), a challenge in treating childhood acute myeloid leukemia (AML), is frequently associated with decreased drug accumulation caused by multidrug transporter MDR1. Doxorubicin, an important anti-AML drug, is a known MDR1 substrate and inducer. Its cytostatic efficacy is thus limited by MDR1 overexpression. A recent study demonstrated cyclooxygenase-2-dependent, prostaglandin E(2) (PGE(2))-mediated regulation of mdr1b expression in primary rat hepatocyte cultures. Cyclooxygenase-2 expression is increased in several malignancies and considered a negative prognostic factor. Our study focused on cyclooxygenase system's impact on drug-induced MDR1 overexpression in AML cells. As a prerequisite, coexpression of MDR1 and cyclooxygenase-2 mRNA in HL-60 cells and primary AML blasts was demonstrated by Northern blot. Interestingly, incubation of AML cells with doxorubicin not only induced functionally active MDR1 overexpression but also mediated increased cyclooxygenase-2 mRNA and protein expressions with subsequent PGE(2) release (determined by flow cytometry, rhodamine123 efflux assay, reverse transcription-polymerase chain reaction, and enzyme-linked immunosorbent assay). After preincubation and subsequent parallel treatment with the cyclooxygenase-2-preferential inhibitor meloxicam, doxorubicin-induced MDR1 overexpression and function were reduced (maximally at 0.1-0.5 microM meloxicam), whereas cytostatic efficacy of doxorubicin in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assays was significantly increased by up to 78 (HL-60) and 30% (AML blasts) after 72 h of doxorubicin treatment. In HL-60 cells, meloxicam-dependent effect on doxorubicin cytotoxicity was neutralized by PGE(2) preincubation. In conclusion, the cyclooxygenase system, especially the cyclooxygenase-2 isoform, might be involved in regulating doxorubicin-induced MDR1 overexpression in AML cells, with PGE(2) seeming to be a mediating factor. Cyclooxygenase inhibitors thus bear promise to overcome MDR in AML and improve therapy.





Eicosanoide und Regulation ihrer Synthese

Differential regulation of cyclo-oxygenase-2 and 5-lipoxygenase-

activating protein (FLAP) expression by glucocorticoids in

monocytic cells.

1Goppelt-Struebe M, 2Schaefer D, 1Habenicht AJ.

Br J Pharmacol. 1997 Oct;122(4):619-24.

1Department of Internal Medicine IV, University of Erlangen-Nürnberg, Erlangen, 2Medical Clinic III, Dept. of Allergology, Universität Erlangen-Nürnberg, Erlangen, Germany Germany.

1. The objective of the present study was to determine the effects of dexamethasone on key constituents of prostaglandin and leukotriene biosynthesis, cyclo-oxygenase-2 (COX-2) and 5-lipoxygenase activating protein (FLAP). The human monocytic cell line THP-1 was used as a model system. mRNA and protein levels of COX-2 and FLAP were determined by Northern and Western blot analyses, respectively. 2. Low levels of COX-2 and FLAP mRNA were expressed in undifferentiated THP-1 cells, but were induced upon differentiation of the cells along the monocytic pathway by treatment with phorbol ester (TPA, 5 nM). Maximal expression was observed after two days. 3. Coincubation of the undifferentiated cells with dexamethasone (10(-9) - 10(-6) M) and phorbol ester prevented induction of COX-2 mRNA, but did not affect the induction of FLAP mRNA. 4. Dexamethasone downregulated COX-2 mRNA and protein in differentiated, monocyte-like THP-1 cells. In contrast, FLAP mRNA and protein were upregulated by dexamethasone in differentiated THP-1 cells. After 24 h, FLAP mRNA levels were increased more than 2 fold. Dexamethasone did not change 5-lipoxygenase mRNA expression. 5. Release of prostaglandin E2 (PGE2) and peptidoleukotrienes was determined in cell culture supernatants of differentiated THP-1 cells by ELISA. Calcium ionophore-dependent PGE2 synthesis was associated with COX-2 expression, whereas COX-1 and COX-2 seemed to participate in arachidonic acid-dependent PGE2 synthesis. Very low levels of peptidoleukotrienes were released from differentiated THP-1 cells upon incubation with ionophore. Treatment with dexamethasone did not significantly affect leukotriene release. 6. These data provide evidence that prostaglandin synthesis is consistently downregulated by glucocorticoids. However, the glucocorticoid-mediated induction of FLAP may provide a mechanism to maintain leukotriene biosynthesis through more efficient transfer of arachidonic acid to the 5-lipoxygenase reaction, in spite of inhibitory effects on other enzymes of the biosynthetic pathway.





ABSRTACTS - KURZÜBERSICHT Eicosanoide und Atemwege

1. Hosemann WG, Baenkler HW, Gunther F. ASA-induced release of histamine from nasal mucous membranes in analgesic intolerance and polyposis nasi. Rhinology 1990 Dec; 28(4): 231-238.

2. Baenkler HW, Schäfer D, Hosemann W. Eicoanoid pattern in nasal polyps and nasal mucosa. Rhinology 1996; 34: 166-170.

3. SchäferD, Lindenthal U, Wagner M, Bölcskei PL, Baenkler HW. Effect of prostaglandin E2 on eicosanoid release by human bronchial biopsy specimens from normal and inflamed mucosa. Thorax 1996; 51: 919-932.

4. Schäfer D, Schmid M, Göde U, Baenkler HW. Dynamics of eicosanoids in peripheral blood cells during bronchial provication in aspirin-intolerant asthmatics. Eur Respir J, 1999; 13: 638-646.

5. Schmid M, Göde U, Schäfer D, Wiegand ME. Arachidonic acid metabolism in nasal tissue and peripheral blood cells in aspirin intolerant asthmatics. Act Otholaryngol (Stockh) 1999; 199: 277-280.

6. Kaldenbach T, Schäfer D, Gosepath J, Bittinger F, Klimek L, Mann WJ. Die Bedeutung eosinophiler Granulocyten in Beziehung zu Allergie und Aspirin-Intoleranz bei Patienten mit Sinusitis polyposa. Laryngo Rhino Otol 1999; 78: 429-434.

7. Gosepath J, Hoffmann F, Schäfer D, Amedee RG, Mann WJ. Aspirin Intolerance in patients with chronic sinusitis. ORL 1999; 61: 146-150.

8. Mohammadian P, Schaefer D, Hummel T, Kobal G. Experimental induced nasal irritation. Rhinology 1999; 37: 176-178.

9. Gosepath J, Schäfer D, Amedee RG, Mann WJ. Individual monitoring of aspirin desensitization. Arch Otolaryngol Head Neck Surg 2001; 127: 316-321.

10. Grundmann T., Töpfer M.. Leukotrienrezeptorantagonisten zur Rezidivprophylaxe bei der ASS-assoziierten Polyposis - Erste klinische Ergebnisse zur Wirkung auf entzündliche





Gewebeprozesse. Laryngorhinootologie 2001; 80: 576-582.

11. Gosepath J, Schäfer D, Mann WJ. Analgetika-Intoleranz: Langzeitergebnisse bis zu 3 Jahren be adaptativer Desaktivierung mit einer täglichen Erhaltungsdosis von 100 mg Aspirin. Laryngo-Rhino-Otol 2002; 81: 732-738.

12. Haxel BR, Schäfer D, Klimek L, Mann WJ. Prostaglandin E2 activates the ciliary beat frequency of cultured human nasal mucosa via second messanger cyclic adenosine monophosphate. Eur Arch Otorhinolaryngol 2001; 258: 230-235.

13. Hecksteden K, Schäfer D, Stuck BA, Klimek L, Hörmann K. Diagnosk des Analgetika-Intoleranz-Syndroms mittels funktioneller Zelltestung (analgetika-Intoleranz-Test: AIT). Allergologie 2003; 26: 263-271.

14. Kuehnemund M, Ismail C, Brieger J, Schaefer d, Mann WJ. Untreated chronic Rhinosinusitis: a comparison of symptoms and mediator profiles. The Laryngoscope, 2004;114:561-565.

15. Eger G, D Schäfer, HW Baenkler. Analgetikasensitive Atemwegserkrankung: Diagnostik, Risikoanalyse und Therapieplanung mittels FET Pneumologie 2004; 58

Eicosanoide und Haut 1. Sauer SK, Schafer D, Kress M, Reeh PW.

Stimulated prostaglandin E2 release from rat skin, in vitro Life Sci. 1998; 62(22): 2045-255.

Eicosanoide und Magen-Darm-Kanal 1. Baenkler HW, Zeus J, Schenk J, Schäfer D.

Abnormal eicosanoid pattern by blood leukocytes in gastroduodenal ulcer. Med Sci Monit. 2004 Oct;10(10):CR557-62. Epub 2004 Sep 23.

Eicosanoide und Nervensystem 1. Schneider N, Lanz S, Ramer R, Schaefer D, Goppelt-Struebe M.

Up-regulation of cyclooxygenase-1 in neuroblastoma cell lines by retinoic acid and corticosteroids. J Neurochem. 2001 Apr;77(2):416-24.





2. Ionac M, Schaefer D, Geishauser M. Thromboxane A2 release in ischemia and reperfusion of free flaps in rats, studied by microdialysis. J Reconstr Microsurg. 2001 Feb;17(2):139-43.

Eicosanoide und Zahnheilkunde 1. Willershausen B, Schafer D, Pistorius A, Schulze R, Mann W.

Influence of resin-based restoration materials on cytotoxicity in gingival fibroblasts Eur J Med Res. 1999 Apr 27; 4(4): 149-155.

2. Willershausen B, Marroquin BB, Schafer D, Schulze R. Cytotoxicity of root canal filling materials to three different human cell lines. J Endod. 2000 Dec;26(12):703-7.

3. Willershausen B, Briseno b, Schulz R, Schäfer D. Untersuchungen zur Zytotoxicität von Wurzelkanalfüllmaterialien. Dtsch Zahnärztl Z 2000;55:252-256.

Eicosanoide und Tumortherapie 1. Ziemann C, Schafer D, Rudell G, Kahl GF, Hirsch-Ernst KI.

The cyclooxygenase system participates in functional mdr1b overexpression in primary rat hepatocyte cultures Hepatology. 2002 Mar; 35(3): 579-588

2. Puhlmann U, Ziemann C, Ruedell G, Vorwerk H, Schaefer D, Langebrake C, Schuermann P, Creutzig U, Reinhardt D Impact of the cyclooxygenase system on doxorubicin-induced functional multidrug resistance 1 overexpression and doxorubicin sensitivity in acute myeloid leukemic HL-60 cells. J Pharmacol Exp Ther. 2005 Jan;312(1):346-54. Epub 2004 Oct 22.

Eicosanoide und Regulation ihrer Synthese 1. Goppelt-Struebe M, Schaefer D, Habenicht AJ..

Differential regulation of cyclo-oxygenase-2 and 5-lipoxygenase-activating protein (FLAP) expression by glucocorticoids in monocytic cells. Br J Pharmacol. 1997 Oct;122(4):619-24.


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