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Polyamine, insbesondere Spermidin und Spermin, sind wichtige Regulatoren grundlegender Zellfunktionen, die darüber hinaus spezifische Funktionen im Gehirn von Säugetieren erfüllen. Dementsprechend sind Polyamine sowie deren Synthese- und Abbauenzyme in Neuronen und Gliazellen in den jeweiligen Gehirnarealen unterschiedlich verteilt. Der Vorläufer für die Spermidin /Spermin-Synthese, das Diamin Putrescin, kann über zwei unterschiedliche Reaktionswege synthetisiert werden: den „klassischen“ und den „alternativen“ Polyaminstoffwechselweg. Während auf dem klassischen Weg Putrescin durch Abspaltung von Harnstoff und Kohlendioxid, katalysiert durch Arginase und Ornithindecarboxylase, entsteht, kann der Vorläufer der Spermidin/Spermin- Synthese auf dem alternativen Weg, nach Abspaltung von zunächst CO2 durch Arginindecarboxylase und nachfolgend Harnstoff durch Agmatinase, gebildet werden. Um das Expressionsmuster von Arginase im Gehirn der Ratte zu untersuchen, wurde in der vorliegenden Arbeit ein polyklonaler, monospezifischer Antikörper durch Aufreinigung eines gegen Arginase 1 gerichteten Rohserums charakterisiert. Mit Hilfe des gereinigten Antikörpers wurde dann die Expression der Arginase im Cortex cerebri, Hippocampus, Thalamus, Hypothalamus, Mesencephalon, Metencephalon und in den Basalganglien auf zellulärer und subzellulärer Ebene analysiert. Im Gegensatz zu anderen Enzymen des Polyaminstoffwechsels, wie z.B. der nachgeschalteten Enzyme Agmatinase und Spermidinsynthase, sind die vorgeschalteten Enzyme Arginase und Arginindecarboxylase in den Neuronen des Rattenhirns deutlich weiter verbreitet. Um Rückschlüsse auf die Bedeutung der verschiedenen Wege der Polyaminsynthese im Gehirn der Ratte ziehen zu können, wurden die Verteilungsmuster der Arginase mit denen von Agmatinase, Spermidinsynthase und Arginindecarboxylase verglichen. Die beobachteten Verteilungsmuster deuten darauf hin, dass der alternative Syntheseweg wahrscheinlich primär der Bildung von Agmatin und nicht Putrescin dient. Dagegen dürfte die Synthese von Spermidin/Spermin vermutlich hauptsächlich über den klassischen Weg erfolgen., Polyamines, namely spermidine and spermine, are important regulators of basic cell functions which are also involved with specific functions in the mammalian brain. Accordingly, polyamines as well as the enzymes for polyamine synthesis and degradation are differentially distributed in neurons and glial cells in the respective brain areas. The precursor of spermidine/spermine synthesis, the diamine putrescine, can be synthesised via two different pathways, refered to as the "classical" and the "alternative" pathway. While in the classical pathway urea and carbon dioxide are removed from the amino acid arginine catalysed by arginase and ornithine decarboxylase, yielding putrescine, the alternative pathway, by first removing carbon dioxide by arginine decarboxlyase and then urea by agmatinase, may also supply the precursor for spermidine/spermine synthesis. To examine the expression pattern of arginase in the rat brain, in this study a polyclonal monospecific antibody was characterized by purification of a raw serum raised against arginase 1. Using the purified antibody, the expression of arginase was analysed in cerebral cortex, hippocampus, thalamus, hypothalamus, mesencephalon, metencephalon and in the basal ganglia at a cellular and subcellular level. In contrast to other polyamine metabolic enzymes, such as the downstream enzymes agmatinase and spermidine synthase, the pathway entry enzymes arginase and arginine decarboxylase are clearly more widely distributed in rat brain neurons. In order to evaluate the significance of either the classical or the alternative pathway for polyamine synthesis in rat brain, the resulting labeling patterns for arginase were compared with those for agmatinase, spermidine synthase and arginine decarboxylase. With regard to the observed distribution patterns, it seems likely that the synthesis of agmatine rather than putrescine may be the main purpose of the alternative pathway of polyamine synthesis. By contrast, spermidine/spermine synthesis may mainly be fueled by the classical pathway.

Im Methioninzyklus wird Methylthioadenosin (MTA), das Nebenprodukt der Polyamin-, Ethylen- und Nicotianaminbiosynthese, zu Methionin recycelt. In dieser Arbeit wurden zum Einen mögliche Regulationen des Methioninzyklus durch die Polyaminbiosynthese in Arabidopsis thaliana untersucht. Arabidopsis besitzt zwei Gene für die MTA-Nukleosidase (MTN; At4g38800, At4g34840), welche den Eingangsschritt des Methioninzyklus katalysiert. T-DNA-Insertionsmutanten von MTN1 zeigten bei Anzucht auf MTA als Schwefelquelle ein stark reduziertes Wachstum. mtn1-1/mtn2-1 Doppelmutanten besaßen eine verlängerte vegetative Wachstumsphase und waren steril. Es wurde bereits gezeigt, dass Spermidin in der Lage ist das reduzierte Keimlingswachstum und die Infertilität der Doppelmutante zu komplementieren. In dieser Arbeit konnte das reduzierte Wurzelwachstum der mtn1 Mutanten durch Spermin komplementiert werden, obwohl die internen Spermingehalte in den Wurzeln auf MTA als Schwefelquelle erhöht waren. Es ist möglich, dass die Inhibierung von Enzymen durch MTA bei dem reduzierten Wurzelwachstum der mtn1 Mutanten eine Rolle spielt. MTA ist in der Lage Enzyme wie die Spermidin-Synthase in vitro zu inhibieren. Um den Zusammenhang von Wurzelwachstum und Polyaminen weiter zu untersuchen, wurden Loss-of-function-Mutanten verwendet. Mutanten der Spermidin-Synthase (At1g23820, At1g70310), Spermin-Synthase (At5g53120), Thermospermin-Synthase (At5g19530) und zwei Acyl-Transferasen (At2g23510, At2g25150) zeigten keinen, den mtn1 Mutanten ähnlichen Phänotyp. Die Inhibierung dieser Enzyme der Polyaminbiosynthese scheint nicht für das reduzierte Wachstum der mtn1 Mutanten verantwortlich zu sein. Über welche Mechanismen Spermin das Wurzelwachstum verändert ist noch nicht verstanden. In dieser Arbeit wurden außerdem die Regulation des Methioninzyklus durch die Ethylenbiosynthese und den Ethylensignalweg untersucht. Es wurde gezeigt, dass Ethylen zu einer Reduktion der MTN-Aktivität über die Reduktion der MTN1 Proteinmenge führt. Die Reduktion von MTN1 scheint weder transkriptionell noch über den Proteinabbau reguliert zu sein. Analysen der Ethylenmutanten ethylene insensitive2 (ein2), ethylene overproducer3 (eto3) und constitutive triple response1 (ctr1) zeigten, dass die Reduktion der MTN1 Proteinmenge nicht über die Ethylenproduktion, sondern über den Ethylensignalweg reguliert wird. Es wäre denkbar, dass der Ethylensignalweg ein Protein reguliert, welches direkt mit MTN1 interagiert um die Proteinmenge zu reduzieren. Protein Interaktion mit MTN1 wurde bereits für CALCINEURIN B-LIKE3 (CBL3) gezeigt. Diese Interaktion ist Kalzium-abhängig und führt zu einer Reduktion der MTN1-Aktivität. In dieser Arbeit wurde untersucht, ob die Reduktion der MTN-Aktivität durch Ethylen über Kalzium-abhängige Interaktion CBL3 reguliert wird. Kalzium führte zu einer Reduktion der MTN-Aktivität, unabhängig von Ethylen. Kalzium-abhängige Interaktion mit CBL3 ist nicht der Grund für die reduzierte MTN-Aktivität in Anwesenheit von Ethylen. Der genaue Mechanismus zur Regulation von MTN1 über den Ethylensignalweg muss noch weiter analysiert werden. The Met cycle salvages the reduced sulfur group from methylthioadenosine (MTA) the by-product of polyamine, ethylene and nicotianamine synthesis. In one part of this thesis Arabidopsis thaliana was employed to study regulation of the Met cycle by the polyamine biosynthesis. In Arabidopsis two genes encode MTA nucleosidase (MTN; At4g38800, At4g34840), which catalyzes the first committed step in the Met cycle. T-DNA-Insertion mutants of MTN1 grown on MTA as sulfur source showed severely reduced growth. mtn1-1/mtn2-1 double mutants had a prolonged vegetative stage and were sterile. It was already shown that spermidine complements retarded growth of mtn1 seedlings and infertility of mtn12/mtn2-1 double mutants. In this thesis reduced root growth of mtn1 plants was complemented by spermine, although internal spermine levels were elevated in roots grown on MTA as sulfur source. It is possible that enzyme inhibition by MTA plays a role in reduced root growth of mtn1. MTA is able to inhibit enzymes like spermidine synthase in vitro. To further study root growth in relation to polyamines, loss-of-function mutants were employed. Mutants of spermidine synthase (At1g23820, At1g70310), spermine synthase (At5g53120), thermospermine synthase (At5g19530) and two acyl transferases (At2g23510, At2g25150) did not have a phenotype comparable to mtn1 grown on MTA. Disrupting activity of these enzymes does not seem to be responsible for reduced mtn1 growth. It is not yet understood how spermine affects root growth of mtn1 mutants. Furthermore this study was employed to analyze regulation of the Met cycle by the ethylene biosynthesis and the ethylene signaling pathway. In this thesis it was shown that ethylene leads to a reduction of MTN activity by reducing the abundance of MTN1 protein. Neither changes in MTN transcripts nor changes in protein degradation were responsible for reduced MTN1 protein abundance. Studying the ethylene mutants ethylene insensitive2 (ein2), ethylene overproducer3 (eto3) and constitutive triple response1 (ctr1) pointed to the ethylene signaling rather than ethylene synthesis as a cause for MTN1 reduction. It is possible that ethylene signaling regulates a protein which interacts with MTN1 to reduce protein abundance. Protein interaction with MTN1 was shown for CALCINEURIN B-LIKE3 (CBL3). It is a calcium-dependent interaction and leads to reduced MTN1 activity. The probability of calcium-dependent interaction with CBL3 as the cause of ethylene-induced reduction of MTN activity was analyzed. Calcium led to a reduction of MTN activity independent of the presence of ethylene. Calcium-dependent interaction with CBL3 is hence not the cause for reduced MTN-activity in the presence of ethylene. Further studies will have to be done to reveal the mechanism of regulation.

Die PH ist eine lebensbedrohliche Erkrankung, die unbehandelt zum Tode führt. Sie ist durch einen erhöhten pulmonalarteriellen Druck, erhöhten pulmonalvaskulären Widerstand sowie die mit PH einhergehenden Gefäßumbauprozesse charakterisiert, die letztendlich zum Rechtsherzversagen führen können. Da die der Erkrankung zu Grunde liegenden Mechanismen noch nicht vollständig entschlüsselt sind, zielen die derzeitigen Behandlungsmöglichkeiten eher auf die Verbesserung der Symptomatik, nicht jedoch auf eine Beseitigung der eigentlichen Ursachen.Im Modell der Hypoxie-induzierten PH der Maus führt eine normoxische Reexposition nach vollständiger Ausbildung der Erkrankung zu einer Gesundung der Versuchstiere. In der vorliegenden Arbeit wurden zur Identifizierung der in diesem Heilungsprozess involvierten Gene genomweite Analysen an Mäusen durchgeführt. Hierbei wurde die S-Adenosylmethionindecarboxylase 1 (AMD-1), ein Enzym der Polyaminsynthese, als ein äußerst stark regulierter Kandidat im Prozess des Revertierens der Hypoxie-induzierten PH gefunden. AMD-1 stellte einerseits eines der am stärksten erniedrigten Gene bei Reoxygenierung dar und andererseits eines der am stärksten erhöhten Gene unter chronisch hypoxischen Bedingungen.Durch in situ Hybridisierung wurde die vermehrte Expression von AMD-1 unter pathophysiologischen Bedingungen einer PH sowohl bei der Maus als auch in humanem Material bestätigt. Des Weiteren zeigte sich eine Lokalisation von AMD-1 in pulmonalen Gefäßen, dem Ort der Umbauprozesse bei PH. Interessanterweise wurde AMD-1 nur in pumonalen Gefäßen unter hypoxischen Bedingungen vermehrt exprimiert und nicht in systemischen Gefäßen, was im Hinblick auf eine pharmakologische Intervention eine gewisse Selektivität für die Lunge versprechen könnte. Unter chronischer hypoxischer Exposition der heterozygoten AMD-1-Knockout-Tiere zeigte sich ein präventiver Phänotyp, der in Verringerung der PH resultierte. Im Gegensatz dazu zeigte eine verminderte Expression der Ornithindecarboxylase 1 (ODC-1), dem zweiten geschwindigkeitsbestimmendem Enzym der Polyaminsynthese neben AMD-1, keinen positiven Effekt auf die Hypoxie-induzierten PH.Des Weiteren konnte diese Arbeit zeigen, dass die Hypoxie-induzierte PH durch vermehrte Proliferation von Zellen der Gefäßwand gekennzeichnet ist, die durch Reoxygenierung normalisiert wurde. Eine verminderte AMD-1-Expression sowohl in vivo als auch in isolierten humanen pulmonalarteriellen glatten Muskelzellen führte zu einer verminderten Proliferation und induzierte gleichzeitig Apoptose. Schließlich konnte in der vorliegenden Arbeit durch computergestützte Analysen und molekularbiologische Methoden der Signalweg aufgeklärt werden, der über EGF-Erk1/2-Egr1 zu einer erhöhten AMD-1-Expression unter hypoxischen Bedingungen führt. Darüberhinaus konnten die Untersuchungen zeigen, dass AMD-1 für die Phosphorylierung der Phospholipase C-γ1 (PLC-γ1) in humanen pulmonalarteriellen glatten Muskelzellen sowie in der Mauslunge notwendig ist. PLC-γ1 ist ein wichtiges Signalmolekül in der Regulation der Calciumhomöostase und somit ein möglicher, wichtiger Effektor des Gefäßumbauprozesses. Zusammengefasst wurde in dieser Studie über in vivo und in vitro Untersuchungen gezeigt, dass AMD-1 den Gefäßumbauprozess bei pulmonaler Hypertonie in vivo steuern kann und dass genomweite Studien, insbesondere im Prozess der Revertierung einer PH im chronisch hypoxischen Mausmodell geeignet sind, neue essentielle Kandidaten der PH-Pathogenese, die auch Ziel einer Therapie sein könnten, zu identifizieren., PH is a life-threatening disease leading to death if untreated. This disease is characterized by an elevated pulmonary arterial pressure, pulmonary vascular remodeling and an increased pulmonary vascular resistance, ultimately leading to right heart failure. Since the underlying mechanisms of the disease are not fully understood yet, the actually available treatment strategies are rather symptomatic than targeted to resolve the cause of the disease.In the mouse model of hypoxia-induced PH, reexposure to normoxia after full establishment of the disease induces full recovery from PH. In the present study, a genome wide screening was performed to identify genes involved this recorvery process. In this analysis, s-adenosylmethionine decarboxylase 1 (AMD-1), an enzyme involved in polyamine synthesis, was found as an extremely strong regulated candidate in the reversal of hypoxia-induced PH. On the one hand, AMD-1 displayed one of the strongest downregulated genes during reoxygenation, on the other hand, AMD-1 was one of the strongest upregulated genes upon chronic hypoxic exposure.By applying in situ hybridization increased AMD-1 expression was confirmed under pathophysiological conditions of PH in mice as well as in human IPAH samples. Furthermore, localization of AMD-1 was demonstrated in pulmonary vessels, the site of vascular remodeling processes in PH. Interestingly, enhanced AMD-1 expression under hypoxic conditions was only detected in pulmonary vessels in contrast to systemic vessels. This might predict at least some selectivity for the pulmonary circulation when applying pharmacological intervention.Under chronic hypoxic conditions AMD-1+/- mice displayed a preventive phenotyp, resulting in a reduced PH. In contrast, diminished expression of ornithine decarboxylase 1 (ODC-1), the second rate-limiting enzyme of the polyamine synthesis next to AMD-1, did not display preventive effects on hypoxia-induced PH.Furthermore, in this study it was demonstrated that the hypoxia-induced PH was accompanied by increased proliferation of cells from the vessel wall, which was normalized by reoxygenation. Decreased AMD-1 expression in vivo and in vitro led to decreased proliferation and augmented apoptosis. Moreover, by applying computational based analysis and molecular biological methods, a signaling cascade leading to increased AMD-1 expression under hypoxia via EGF-Erk1/2-Egr1 was identified. Furthermore, it was verified that AMD-1 is required for the phosphorylation of phospholipase C-γ1 (PLC-γ1) in hPASMCs and in the mouse lung. PLC-γ1 is a crucial signaling molecule in the regulation of calcium homeostasis and therefore a possible essential effector of vascular remodeling processes. Taken together, it was shown by in vivo and in vitro approaches, that AMD-1 can regulate the vascular remodeling process in pulmonary hypertension in vivo and that genomewide studies, particularly during the process of reversal of PH in the mouse model of chronic hypoxia, are feasible to identify new, essential candidates in the pathogenesis of PH which may also can serve as a possible target for the pharmacotherapy of PH.

Die PH ist eine lebensbedrohliche Erkrankung, die unbehandelt zum Tode führt. Sie ist durch einen erhöhten pulmonalarteriellen Druck, erhöhten pulmonalvaskulären Widerstand sowie die mit PH einhergehenden Gefäßumbauprozesse charakterisiert, die letztendlich zum Rechtsherzversagen führen können. Da die der Erkrankung zu Grunde liegenden Mechanismen noch nicht vollständig entschlüsselt sind, zielen die derzeitigen Behandlungsmöglichkeiten eher auf die Verbesserung der Symptomatik, nicht jedoch auf eine Beseitigung der eigentlichen Ursachen. Im Modell der Hypoxie-induzierten PH der Maus führt eine normoxische Reexposition nach vollständiger Ausbildung der Erkrankung zu einer Gesundung der Versuchstiere. In der vorliegenden Arbeit wurden zur Identifizierung der in diesem Heilungsprozess involvierten Gene genomweite Analysen an Mäusen durchgeführt. Hierbei wurde die S-Adenosylmethionindecarboxylase 1 (AMD-1), ein Enzym der Polyaminsynthese, als ein äußerst stark regulierter Kandidat im Prozess des Revertierens der Hypoxie-induzierten PH gefunden. AMD-1 stellte einerseits eines der am stärksten erniedrigten Gene bei Reoxygenierung dar und andererseits eines der am stärksten erhöhten Gene unter chronisch hypoxischen Bedingungen. Durch in situ Hybridisierung wurde die vermehrte Expression von AMD-1 unter pathophysiologischen Bedingungen einer PH sowohl bei der Maus als auch in humanem Material bestätigt. Des Weiteren zeigte sich eine Lokalisation von AMD-1 in pulmonalen Gefäßen, dem Ort der Umbauprozesse bei PH. Interessanterweise wurde AMD-1 nur in pumonalen Gefäßen unter hypoxischen Bedingungen vermehrt exprimiert und nicht in systemischen Gefäßen, was im Hinblick auf eine pharmakologische Intervention eine gewisse Selektivität für die Lunge versprechen könnte. Unter chronischer hypoxischer Exposition der heterozygoten AMD-1-Knockout-Tiere zeigte sich ein präventiver Phänotyp, der in Verringerung der PH resultierte. Im Gegensatz dazu zeigte eine verminderte Expression der Ornithindecarboxylase 1 (ODC-1), dem zweiten geschwindigkeitsbestimmendem Enzym der Polyaminsynthese neben AMD-1, keinen positiven Effekt auf die Hypoxie-induzierten PH. Des Weiteren konnte diese Arbeit zeigen, dass die Hypoxie-induzierte PH durch vermehrte Proliferation von Zellen der Gefäßwand gekennzeichnet ist, die durch Reoxygenierung normalisiert wurde. Eine verminderte AMD-1-Expression sowohl in vivo als auch in isolierten humanen pulmonalarteriellen glatten Muskelzellen führte zu einer verminderten Proliferation und induzierte gleichzeitig Apoptose. Schließlich konnte in der vorliegenden Arbeit durch computergestützte Analysen und molekularbiologische Methoden der Signalweg aufgeklärt werden, der über EGF-Erk1/2-Egr1 zu einer erhöhten AMD-1-Expression unter hypoxischen Bedingungen führt. Darüberhinaus konnten die Untersuchungen zeigen, dass AMD-1 für die Phosphorylierung der Phospholipase C-γ1 (PLC-γ1) in humanen pulmonalarteriellen glatten Muskelzellen sowie in der Mauslunge notwendig ist. PLC-γ1 ist ein wichtiges Signalmolekül in der Regulation der Calciumhomöostase und somit ein möglicher, wichtiger Effektor des Gefäßumbauprozesses. Zusammengefasst wurde in dieser Studie über in vivo und in vitro Untersuchungen gezeigt, dass AMD-1 den Gefäßumbauprozess bei pulmonaler Hypertonie in vivo steuern kann und dass genomweite Studien, insbesondere im Prozess der Revertierung einer PH im chronisch hypoxischen Mausmodell geeignet sind, neue essentielle Kandidaten der PH-Pathogenese, die auch Ziel einer Therapie sein könnten, zu identifizieren. PH is a life-threatening disease leading to death if untreated. This disease is characterized by an elevated pulmonary arterial pressure, pulmonary vascular remodeling and an increased pulmonary vascular resistance, ultimately leading to right heart failure. Since the underlying mechanisms of the disease are not fully understood yet, the actually available treatment strategies are rather symptomatic than targeted to resolve the cause of the disease. In the mouse model of hypoxia-induced PH, reexposure to normoxia after full establishment of the disease induces full recovery from PH. In the present study, a genome wide screening was performed to identify genes involved this recorvery process. In this analysis, s-adenosylmethionine decarboxylase 1 (AMD-1), an enzyme involved in polyamine synthesis, was found as an extremely strong regulated candidate in the reversal of hypoxia-induced PH. On the one hand, AMD-1 displayed one of the strongest downregulated genes during reoxygenation, on the other hand, AMD-1 was one of the strongest upregulated genes upon chronic hypoxic exposure. By applying in situ hybridization increased AMD-1 expression was confirmed under pathophysiological conditions of PH in mice as well as in human IPAH samples. Furthermore, localization of AMD-1 was demonstrated in pulmonary vessels, the site of vascular remodeling processes in PH. Interestingly, enhanced AMD-1 expression under hypoxic conditions was only detected in pulmonary vessels in contrast to systemic vessels. This might predict at least some selectivity for the pulmonary circulation when applying pharmacological intervention. Under chronic hypoxic conditions AMD-1+/- mice displayed a preventive phenotyp, resulting in a reduced PH. In contrast, diminished expression of ornithine decarboxylase 1 (ODC-1), the second rate-limiting enzyme of the polyamine synthesis next to AMD-1, did not display preventive effects on hypoxia-induced PH. Furthermore, in this study it was demonstrated that the hypoxia-induced PH was accompanied by increased proliferation of cells from the vessel wall, which was normalized by reoxygenation. Decreased AMD-1 expression in vivo and in vitro led to decreased proliferation and augmented apoptosis. Moreover, by applying computational based analysis and molecular biological methods, a signaling cascade leading to increased AMD-1 expression under hypoxia via EGF-Erk1/2-Egr1 was identified. Furthermore, it was verified that AMD-1 is required for the phosphorylation of phospholipase C-γ1 (PLC-γ1) in hPASMCs and in the mouse lung. PLC-γ1 is a crucial signaling molecule in the regulation of calcium homeostasis and therefore a possible essential effector of vascular remodeling processes. Taken together, it was shown by in vivo and in vitro approaches, that AMD-1 can regulate the vascular remodeling process in pulmonary hypertension in vivo and that genomewide studies, particularly during the process of reversal of PH in the mouse model of chronic hypoxia, are feasible to identify new, essential candidates in the pathogenesis of PH which may also can serve as a possible target for the pharmacotherapy of PH.

Die Unterbrechung des koronaren Blutflusses führt zum Myokardinfarkt. Die am erfolgreichsten durchzuführende Intervention stellt die frühzeitige Reperfusion von Blut dar. So kann es zu einem Überleben der Zellen und einer Funktionserhohlung kommen. Trotz der erneuten Perfusion und dem Überleben des Myokards, welches durch Kontraktionsaktivität gemessen werden kann, kommt es zu neuem Stress: Der sogenannte Reperfusionsstress. Dieser ist charakterisiert durch ein Missverhältnis von Ionen, z.B. einer Anhäufung von H+ und Ca2+, sowie einer Neuregulation zellulärer Gene wie z.B. das pro-apoptotische Gen bax und das anti-apoptotische bcl-2. Diese Prozesse führen u.a. zu hypertrophem Wachstum und myokardialer Fibrose und werden als Remodeling bezeichnet. Es ist bekannt, dass Polyamine in der Funktionserhohlung des post-ischämischen Herzens eine wichtige Rolle einnehmen. Die Aufgabe dieser Studie bestand darin, den Effekt einer Aktivierung und Inhibiereung der Ornithinedecarboxylase (ODC), welche das geschwindigkeitsbestimmende Enzym der Polyaminbiosynthese darstellt, bezogen auf die Funktionserhohlung sowie die frühe Induktion von Apoptose oder hypertrophem Wachstum in Kardiomyozyten zu untersuchen. Aus diesem Grund wurden Rattenherzen einer 45 minütigen Nullfluss-Ischämie mit anschließender 120 minütiger Reperfusion unterzogen. Die Kontrollen wurden für 165 min konstant unter normoxen Bedingungen perfundiert. Die ODC wurde durch die 10 minütige Zugabe von Isoprenalin (10nM), 30 min nach erfolgter Reperfusion aktiviert und durch Difluoromethylornithin (DFMO) inhibiert. Als Funktion des Herzens wurde der linksventrikulär entwickelte Druck angenommen. (LVDP). Die Expression der ODC, bcl-2, bax sowie ANF beider Ventrikel wurde durch die real-time-PCR nach 120 min erfolgter Reperfusion bestimmt. Diese Studie konnte verifizieren, das die Inhibition der ODC Aktivität die Funktionserhohlung der ischämisch-reperfundierten Herzen verschlechterte, wohingegen sich bei den normoxisch perfundierten Herzen keine Änderungen zeigten. Die Induktion der ODC hatte keinen Effekt auf die Funktionserhohlung. Obwohl es zu einer Steigerung der Expression des pro-apoptotischen Gens bax im post-ischämischen Herzen kam (nicht bei den normoxen Herzen), überwog die anti-apoptotische Genexpression durch bcl-2. Darüberhinaus führte die Induktion der ODC zu einer Zunahme von ANF unter post-ischämischen Bedingungen. Diese Ergebnisse zeigten sich in beiden Ventrikeln, wohingegen die Induktion von bcl-2 und ANF im rechten Ventrikel weniger von der ODC Aktivierung abhängig war. Die Resultate dieser Studie belegen den vielversprechenden Ansatz neuer protektiver Strategien des post-ischämischen Herzens, wie den hier untersuchten Einfluss des Polyaminstoffwechsels. Diese Erkenntnisse könnten für neue Wege in der Kardiologie oder Herzchirurgie sorgen. Interruption of coronary perfusion results in myocardial infarction. The early reperfusion of blood flow is the most successful intervention leading to cell survival and functional recovery. However, in spite of the restoration of blood flow and thus the myocardial survival, which is measurable by its contractile activity, the heart is confronted by new stress: The so-called reperfusion injury. This is characterized by a cytosolic disproportion of ions like an accumulation of H+ or Ca2+ and a change concerning the regulation of cellular genes such as the pro-apoptotic gene bax or anti-apoptotic gene bcl-2. These conditions lead among other things to hypertrophic growth and myocardial fibrosis which is termed remodeling. Polyamines are known to play an important role in the functional recovery of the post-ischemic heart. The aim of this study was to investigate the effect of an activation or inhibition of ornithine decarboxylase (ODC), which is the rate-limiting enzyme for polyamines, on the functional recovery and the early induction of apoptotic or hypertrophic pathways in rat cardiomyocytes. For this reason rat hearts were exposed to 45 min no-flow ischemia and 120 min reperfusion, whereas controls were constantly perfused for 165 min under normoxic conditions. ODC was activated by application of isoprenaline (10nM) ,starting 30 min after the onset of reperfusion and lasting for 10 min, and inhibited by application of difluoromethyl ornithine (DFMO). The heart function was determined as left ventricular developed pressure (LVDP). The expression of ODC, bcl-2, bax and ANF were measured by real time RT-PCR in both ventricles at the end of 120 min reperfusion. This study could verified that inhibition of ODC activity worsened the functional recovery of the ischemia-reperfused hearts but did not modify basal function of normoxic perfused hearts. Induction of ODC had no effect on the functional recovery. Although it induced the expression of the pro-apoptotic gene bax in post-ischemic hearts (not in normoxic hearts) the anti-apoptotic gene expression of bcl-2 was leading. Furthermore the induction of ODC lead to an increase of ANF in post-ischemic conditions. These findings could be demonstrated in both vebtricles, whereas the induction of bcl-2 and ANF was less dependent on ODC activation in the right ventricle. The results of this study shows that taking influence of polyamine metabolism is a promising approach concerning new protctive strategies for the post-ischemic heart. These findings could modify new approaches in cardiology or in cardiac surgery.

[no abstract]

Numerous detection systems are available for measuring halitosis. In order to examine their agreement, a study was conducted comparing four selected measuring methods in 100 subjects (52 females, 48 males; mean age: 25 years). Organoleptic halitosis measurement was carried out by an odor judge, and compared with instrumental halitosis measurement by sulfide monitoring using Halimeter, Fresh Kiss, and Halitox (halitosis linked toxin detection assay), with which both VSC (volatile sulphur compounds) and polyamines can be detected. The results show that the values recorded by the Halimeter correlated best with the organoleptic assessment and the least with the results of Fresh Kiss.

In dieser Arbeit wurden die potenziell bleichaktiven Salicylderivate der linearen, primären Polyamine 1,2,3-Triaminopropan (trap), 1,2R,3S,4-Tetraaminobutan (etab) und 1,2S,3S,4—Tetraaminobutan (ttab) durch Kondensation mit Salicyl-2-carbaldehyd und anschließender Reduktion der isolierten Iminintermediate synthetisiert und charakterisiert. Die Pyridlyderivate wurden analog durch Alkylierung mit Pyridin-2-carbaldehyd hergestellt. Die Pyrimidinliganden 6-(4-(4-Methyl-1,4-diazacylohexan)pyridin-2-yl)-2-(pyridin-2-yl)pyrimidin-4-ol (C127) und 6-(4-(4,4-Dimethyl-1,4-diazacylohexan)pyridin-2-yl)-2-(pyridin-2-yl)pyrimidin-4-ol (C128) wurden von der CIBA GRENZACH GmbH synthetisiert und zur Untersuchung der Eigenschaften in Lösung zur Verfügung gestellt. Das Koordinationsverhalten der bleichaktiven Liganden wurde in wässriger Lösung mit den Übergangsmetallen Kupfer(II), Eisen(II)/(III), Mangan(II) und Zink(II) in verschiedenen Metall zu Ligand Verhältnissen bestimmt. Hierbei zeigte sich im Falle des Mangan(II) eine deutliche Tendenz Metall-Ligand-Biskomplexe auszubilden sowie überraschend hohe Stabilitäts-konstanten der Liganden mit Zink(II). Um die Teilchenverteilungen den realen Bedingungen in einer Waschflotte anzunähern wurde mit den zuvor experimentell bestimmten Stabilitäts-konstanten eine vereinfachte Flotte simuliert. Diese berücksichtigte die Einflüsse von Härtebildnern, Fremdionen, den pH-Wert und reale Konzentrationsverhältnisse. Das Verhalten der Salicyl-, respektive der Pyridylderivate der linearen, primären Polyamine wurde mit verschieden Übergangsmetallen bestimmt. Die Titrationsexperimente zeigten eine hohe Tendenz der Ligandsysteme tautomere Strukturen auszubilden, welche durch Molecular Modeling Experimente näher untersucht wurden. In this work several ligands with potential laundry bleaching effect have been examined. The salicylic derivatives of the lineary, primary polyamines 1,2,3-Triaminopropane (trap), 1,2R,3S,4-Tetraaminobutane (etab) and 1,2S,3S,4-Tetraaminobutane (ttab) have been synthesized by condensation with salicyl-2-carbaldehyde followed by reduction of the isolated imineintermediates. The corrseponding pyridlyderivatives were synthesized similarly by use of pyridine-2-carbaldehyde as reagent. The pyrimidineligands 6-(4-(4-Methyl-1,4-diazacylohexan)pyridin-2-yl)-2-(pyridin-2-yl)pyrimidin-4-ol (C127) and 6-(4-(4,4-Dimethyl-1,4-diazacylohexan)pyridin-2-yl)-2-(pyridin-2-yl)pyrimidin-4-ol (C128) were provided by courtesy of CIBA GRENZACH GmbH to explore their behaviour in solution. The complex formation of the bleaching active ligands in aqueous solution have been studied with the transition metals copper(II), iron(II)/(III), manganese(II) and zinc(II) in different ligand to metal ratios. The significant tendency to form bis-species in case of the manganese(II) complexes as well as unexpectet high stability constants of the zinc(II) complexes were discovered. To approximate the calculated species distributions to real conditions in a suds, simulations were performed with consideration of the experimental stability constants. The influences of phosphonates, interfering ions, pH-value and real concentrations have been included. The behaviour of the salicylic- respective pyridlylderivatives of the lineary, primary polyamines in solution, has been examined with different transition metals. The experiments showed a very high tendency to form tautomeric structures, which were examined using molecular modeling techniques.

In dieser Arbeit wird beschrieben, wie Polyamine, gebunden an geladene Stellen der Proteingrenzflächen, als Modulatoren transienter Protein-Protein Interaktionen fungieren können. Die Fähigkeit von Putrescin, Spermidin und Spermin, die Protein-Protein Interaktionen zwischen Adrenodoxin Reduktase (AdR), Adrenodoxin (Adx) und CYP11A1 zu beeinflussen, wurde unter anderem mittels Oberflächenplasmonresonanz und Stopped-Flow analysiert. Die Ergebnisse zeigten modulierende Effekte der Polyamine auf einzelne Interaktionen und auf das gesamte System. Computersimulationen der Polyamine an die Kristallstrukturen von oxidiertem Adx, AdR und dem oxidierten Adx — AdR Komplex, die von Frau M.Sc. S. Eyrisch, Universität des Saarlandes, durchgeführt wurden, deuten darauf hin, dass eine Modulation durch veränderte Protein-Protein Interaktionen möglich ist. Die Docking Experimente schlagen eine bevorzugte Bindestelle an der Region Asp-15, Asp-39 und Asp-41 des ungebundenen Adrenodoxins vor. Vier Adx-Mutanten (AdxD15K, AdxD15N, AdxD39K und AdxD39N) zeigten mit Ausnahme einer erhöhten thermischen Stabilität keine spektroskopischen Unterschiede zum Wildtyp. Die Mutanten verhielten sich in ihren Elektronentransfereigenschaften wie der Wildtyp in Anwesenheit der Polyamine. Somit konnte zum ersten Mal gezeigt werden, dass Polyamine mit unterschiedlichen Komponenten transienter Proteinkomplexe interferieren und Proteinfunktionen kontrollieren können. This work describes the role of polyamines that could efficiently bind to charged spots at protein interfaces as modulators of transient protein-protein interactions. The capability of putrescine, spermidine, and spermine to affect the protein-protein interactions between adrenodoxin reductase (AdR), adrenodoxin (Adx), and CYP11A1 was analysed using techniques like surface plasmon resonance or stopped flow. These studies revealed modulating effects of polyamines on distinct interactions and on the entire system. Computer simulations of the polyamines docked into the crystal structures of oxidized Adx, AdR, and the oxidized Adx — AdR complex performed by M.Sc. S. Eyrisch (Saarland University) suggested that modulation via changed protein-protein interactions appear plausible. On Adx, the docking experiments suggested one favourable high-affinity binding site of polyamines, the region around Asp-15, Asp-39, and Asp-41 of free Adx. Four Adx mutants (AdxD15K, AdxD15N, AdxD39K, and AdxD39N) showed, except a higher thermo-stability, no differences regarding their spectroscopic characteristics to the adrenodoxin wildtype. In their electron transfer properties the mutants were comparable to AdxWT in presence of the polyamines. These findings imply for the first time that small endogenous compounds are capable of interfering with distinct components of transient protein complexes and might control protein functions.

Ausgehend von 1,4-Diazepan-6-amin (daza), 6-Methyl-1,4-diazepan-6-amin (aaz) und cis-3,5-Diaminopiperidin (cis-dapi) wurde durch reduktive Alkylierung eine Reihe von Pyridylmethylderivaten dieser cyclischen Triamine hergestellt. In potentiometrischen und spektrophotometrischen Titrationen wurden die Stabilitätskonstanten der 4- bis 6-zähnigen Liganden mit den Übergangsmetallen Mn2+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+ und Cd2+ bestimmt. Dabei zeichneten sich die durch die Pyridylmethylgruppen azidifierten Liganden durch hohe Komplexstabilitäten vor allem im sauren pH-Bereich aus. Je nach Substitutionsgrad unterscheiden sich die Derivate in ihrer Selektivität bezüglich einer Komplexierung von Cu2+ und Ni2+. In Festkörperstrukturen der Komplexe mit Fe3+, Co3+, Ni2+ und Cd2+ konnte Koordinationszahl 6 mit Koordinationspolyedern zwischen einer oktaedrischen und trigonal-prismatischen Geometrie nachgewiesen werden. Dagegen fand man für Komplexe mit Cu2+ und Zn2+ Koordinationszahl 5 mit Polyedergeometrien zwischen einer trigonalen Bipyramide und einer tetragonalen Pyramide. UV/Vis-spektroskopische Untersuchungen zeigten, dass die Pyridylmethylderivate von daza ein vergleichsweise schwaches Ligandenfeld besitzen. Cyclovoltammetrische Messungen an Mn-, Fe- und Co-Komplexen belegten die Pi-Akzeptorfähigkeit der Chelatoren. Obwohl die Liganden zur Bildung von [LFe(III)(OOH)]-Komplexen befähigt sind, ist deren katalytische Aktivität zur Oxidation von Cycloocten nur gering ausgeprägt. A number of pyridylmethyl derivatives of the cyclic triamines 1,4-diazepan-6-amine (daza), 6-methyl-1,4-diazepan-6-amine (aaz) and cis-3,5-diaminopiperidine (cis-dapi) has been prepared by reductive alkylation. The stability constants of the tetra-, penta- and hexadentate ligands with the divalent transition metals Mn2+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+ and Cd2+ were determined by means of potentiometric and spectrophotometric titrations. All derivatives which are acidified by their pyridylmethyl-pendant arms revealed high complex stabilities especially in acidic solution. Depending on the degree of substitution the ligands show different selectivity for the metals Cu2+ and Ni2+. Solid state structures of complexes with Fe3+, Co3+, Ni2+ and Cd2+ exhibited co-ordination number 6 with distorted octahedral or trigonal prismatic coordination poly-hedra. In contrast, in the crystal structures of complex species with Cu2+ and Zn2+ coordination number 5 with a geometry in-between a trigonal bipyramidal and a square pyramidal arrangement was found. As shown by UV/Vis spectrophotometric measurements the pyridylmethyl derivatives of daza evoke only a weak ligand field compared to related ligands. Cyclovoltammetric studies of complexes with Mn, Fe and Co established the pi-acceptor capability of the chelators. Although the ligands are capable of forming [LFe(III)(OOH)]-species, their catalytic activity in oxidation reactions with cycloocten was only minor characteristic.

New findings on the molecular biology of plasmodia have led to new approaches for the development of chemotherapeutics. Trials to circumvent the relative toxicity of individual substances by a combination of active agents have been successful in several cases, and there will be more of those combinations in the future. An additional advantage of combinations can be the improvement of activity. In some cases, antiparasitic side effects of long known drugs with low toxicity such as imipramine can be used to decrease or overcome resistance. And, last but not least, ideas or in some cases new drugs will come from the traditional medicine.

The impairment of the functions of red blood cells or their destruction during storage can be delayed or even inhibited by oligoamines especially RE 1492 (N,N',N''-Tris-(4-phenylbutyl)benzene-1,3,5-trimethanamine). When citrated whole blood (WB) is stored at 4 degrees C for 7 d half of the red blood cells (RBC) have lost their ability to form rouleaux. Addition of 100 mumols/L RE 1492 maintains 50% reaggregability up to day 28th of storage. When citrated WB is stored at 37 degrees C the reaggregability has declined to 40 percent after 10 h. With 100 mumols/L RE 1492 no reduction of this property is observed up to 48 h. These results are correlated with the maintainance of the discocyte form of RBC and a persistent filtrability of RBC suspensions through a 5 microns microporous membrane. With 100 mumols/L RE 1492 only one fifth of the haemolysis of untreated WB occurs. The efflux of potassium ions from RBC into the blood plasma during a 72 h storage is bisected by RE 1492. The binding of oxygen to RBC remains unchanged.

Sodium oxalate (402 mg) was administered in a single dose to 10 healthy volunteers receiving a controlled diet. Half the group received 3 times 10 g Colestid and the other half 4 times 2 g Andursil. On the 5th day the oxalate load was repeated. Urine was collected within 32 hours following oxalate application in 8 fractions. In each fraction the levels of oxalate, calcium, phosphate and uric acid were determined. The amount of oxalate, phosphate and uric acid measured in the group receiving Colestid was lower in all fractions. Peak excretions of oxalate found in unmedicated volunteers were suppressed following oxalate load. In the group receiving Andursil, only the excretion of phosphate was decreased. The results presented suggest that Colestid may be promising in the prevention of calcium-oxalate-urolithiasis.

The cupriuretic effectivity of tetradentate chelating agents was measured in rats and compared with that of D-penicillamine. The applied chelators are aliphatic and alicyclic tetramines and tetramine-analogous substances with two nitrogen-atoms substituted by oxygen. Urine copper excretion was measured during 24 h (and 6 and the following 18 h, respectively) after single and repeated doses over a period of 5 days. The aliphatic tetramines, triethylenetetramine (TETA, TRIEN) and BE 6184, were the most active substances. Simultaneous application of TETA and D-penicillamine does not induce a summation or potentiation of the copper elimination. After s.c. application TETA seems to be three times more effective than orally. The potency of aliphatic and alicyclic tetramines is discussed on the basis of in vitro data.

The most reliable laboratory test for prostate cancer remains prostatic phosphatase determination. With the spectrophotometric method, however, falsely negative results are to be expected in 40% of stage D lesions. In only one third of patients with localized disease results are correctly positive. This poor specificity and sensitivity can be improved by radioimmunoassay (RIA). Using this technique the prostatogenic isoenzyme is elevated in 50% of stage A and in 80% of stage B carcinoma, suggesting RIA for screening. Erythrocyte sedimentation rate or serum iron and copper are not necessarily of prognostic value. Phosphatase determination of bone marrow aspirates also requires the RIA method if differentiation of stage C and D is to be expected. Serum hormone assays are not yet introduced into routine staging programs. Serum and urinary markers such as CEA, polyamines of LDH isoenzymes are unspecific and of uncertain value in prostatic carcinoma. Measurement of urinary hydroxyproline seems a reliable method for the search of osseous spread; other bone diseases have to be excluded. In patients with prostate cancer laboratory tests still represent adjunctive measures in connection with the clinical diagnostic armamentarium of urologists.

The paper is a review of all available suture materials (absorbable and non-absorbable threads). The tissue reaction to the various sutures is discussed and histologically demonstrated, as well as possible complication. It is described which suture material is best suited for the various tissues and suturing techniques.

The C-reactive serum protein (CRP) is a classical acute-phase reactant of increasing diagnostical value in clinical practice. It is drastically elevated during reactions of inflammation and tissue destruction, and it binds selectively to necrotic cells of inflamed tissue. The phylogenetically rather ancient protein displays a cyclic pentameric symmetry (pentraxin). CRP undergoes specific binding to phosphocholine esters, polycations and galactans (lectin character). Aggregated CRP activates complement, has opsonin properties and interacts with lymphocytes and thrombocytes. Probably CRP is part of an early unspecific protective mechanism, whereby potentially toxic materials can be detected and eliminated.

Determination of polyamines (putrescine, spermidine) can be useful for evaluation of treatment efficacy. The amount of putrescine and spermidine were estimated for 10 patients (breast carcinoma; stage IV; T0--4 N0--3 M1) prior to, during and after different combination chemotherapy (carminomycin-dibromodulcitol; cyclophosphamide-methotrexate-5-fluorouracil). Hydrolysis with hydrochloric acid showed best results. Automated ion exchange chromatography is a sensitive and reproducible method. 6 out of 10 patients did not show both clinical changes and changes in polyamine content comparable to values prior to combination chemotherapy. One example demonstrated both clinical progression and an increase of polyamine content. A correlation between decrease of putrescine and spermidine level and clinical remission were found in 3 out of 10 patients. These results support the usefulness of polyamine determinations to evaluate cancer chemotherapy.

64 hypercholesterolemic diabetics (17 males and 47 females) were treated with Colestipol for 3 years. An average reduction of 29% in serum cholesterol levels was observed after 3 years. An increase in triglyceride levels was observed in several instances. Other laboratory data failed to reveal any deviation from normal ranges. Gastrointestinal complaints and constipation were initially noted with onset of therapy. These side effects resolved with continuation of treatment.

Plasma lipids and lipoproteins were studied in 20 children and adolescents with familial hypercholesterolemia during the administration of two anion exchange resins. All the patients' cholesterol levels had been stabilized by treatment with a cholesterol-lowering diet for at least 12 months. Colestyramin (0.6 g per kg body weight) or Colestipol (0.5 g per kg body weight) were given for eight weeks each in a cross-over design. In 6 children, the study had to be terminated due to gastrointestinal problems. Cholesterol, triglycerides and phospholipids were determined serially in whole serum and in isolated lipoprotein fractions. Apolipoprotein-B was determined by radial immunodiffusion in whole serum and in the LDL-fraction. Total plasma cholesterol and LDL cholesterol were lowered by about 25% from initial values of 290 mg/100 ml and 230 mg/100 ml, respectively and Apo-B was lowered by 20%. Low HDL-cholesterol levels in the patients remained decreased during therapy. Triglyceride and phospholipid levels were not affected by treatment. Efficacy, tolerance and side-effects were similar with both Colestyramin and Colestipol.

The polyamine metabolism is pathologically changed in tumor tissues, and especially putrescine and spermidine demonstrate abnormally high values in kidney, bladder, and prostate cancer. The inductive processes which activate the biosynthetic polyamine enzymes in cancer are completely unknown. Of therapeutic interest is the fact that increased enzyme activities through irreversible inhibitors become significantly reduced, which consequently slows the tumor growth. Experimental therapy, especially in transplantable bladder and prostate cancer, displayed a 50% tumor destruction. In clinical studies using inhibitors of the polyamine biosynthesis, the dose had to be significantly reduced because of expressed toxicity. Additional investigations which tried a combination of reversible and irreversible inhibitors proved a similar antitumor activity, but less severe side effects.

The concentration of histamine, cadaverine, putrescine as well as spermidine and spermine was measured in blood serum of patients with rheumatoid arthritis (RA). In comparison to normals there is no statistically significant difference in the concentration of putrescine and spermidine in blood serum of RA patients. Spermine which could be detected in all samples too did not show distinct abnormalities from healthy subjects. Histamine and cadaverine present in measurable amounts were only in particular cases. A relation between increased histamine and basophile count was not demonstrable. There is apparently no correlation between histidinemia which is reported in the literature for RA patients in blood serum and plasma, and histamine production in the blood.

We compared the manual Polybrene technique to three standard methods (albumin/Coombs, LISS/Coombs, and enzyme/papain) on 113 red cell antibodies. Polybrene identified 8 antibodies of the Rhesus system missed by standard methods, whereas 3 antibodies could only be detected by the standard techniques. Four antibodies were identified only in saline solution at room temperature; 6 were found by use of Polybrene exclusively in the additional Coombs phase. In addition, 61 antibodies were tested by 4 different LISS. No considerable differences in the quality of the various LISS were seen. The manual Polybrene test appears to be suitable for crossmatching and rapid antibody identification in emergency situations.

In 1967 Spock et al. reported on the serum of cystic fibrosis (CF) homozygotes containing a factor altering the coordination of ciliary motion in rabbit tracheal explants. Just in 1967 Mangos et al. found sweat and saliva from CF homozygotes having an inhibitory effect on sodium reabsorption in the rat parotid gland. Since that time the existence of CF specific humoral factors was supposed. Hitherto mainly biological tests (especially tests of ciliary dyskinesia) were used to prove these factors. These tests caused different results which even were doubtful with regard to the existence of CF specific proteins. Recently it is possible to differentiate between proteins with effects of ciliary dyskinesia and a CF specific protein by means of high sensitive biochemical and immunological methods of protein distinction. In future one can expect elucidation of question related to the importance of CF protein in pathogenesis and diagnosis of cystic fibrosis.

Longterm treatment for 5 years of young patients with familial hypercholesterolemia was accompanied by monitoring of plasma levels of calcium, iron, sodium, parathyroid hormone and water- and fat soluble vitamins, since interference of the ion exchange resin with absorption of numerous substances as well as abnormal plasma levels of some of the above had been described in several studies. Treatment was effective in the group with good compliance (lowering of plasma cholesterol at the end of 5 years by 19% and, compared to the control group, by 23%). Adverse drug effects with respect to the above parameters were not found. Plasma levels of carotinoides and vitamin E were elevated in the patients according to elevated concentrations of lipoproteins which are carriers of these vitamins.

The advantages of breast feeding are due to numerous factors i.e. the favourable composition of basic nutritives, the content of nucleotides, polyamines, rare fatty acids and rare oligo- and polysaccharides in human milk, its high content of immunologically active substances like immunoglobulins, fats, glycoproteins, lysozyme and lactoferrin, and the psychologically important opportunity for close sensual contact between mother and infant. Involution of the uterus is greatly accelerated by breast feeding. In case the mother has to take certain drugs her breast feeding can be disadvantageous for the infant. At present, we do not see any reason to recommend discontinuation of breast feeding because of the relatively high DDT content found in human milk, since the concentration observed has shown no toxic effects. Furthermore a decrease of DDT concentration in human milk has been observed recently.

The study comprises 90 diabetic patients with hypercholesterolemia. They received colestipol, a basic anion exchange resin, which binds the bile acids in the intestinal tract and thus prevents its enterohepatic circulation. The constant regeneration of bile acids from cholesterol leads to a decrease of the cholesterol level. A significant reduction of the cholesterol level was achieved in 73 of 85 evaluated cases. The observed triglyceride levels showed a different behaviour. All other laboratory data, especially the blood-sugar level, remained unchanged under the treatment with colestipol. Colestipol was well tolerated and, due to its neutral taste, was readily taken by the patients. Only a few patients suffered from slight constipation which did not lead to discontinuation of the treatment.

The polyamines putrescine, cadaverine, spermidine, spermine, and the amine histamine in whole blood and plasma of 42 patients with malignant melanoma were analysed after cold acid extraction and ion-exchange chromatography by fluorescence detection. Depending on the degree of tumor spreading the patients were grouped into 5 subpopulations; results were statistically compared with the values obtained from a normal population. The mean values of polyamine concentrations in the patient groups were generally not distinguishable from the mean values of the controls. Solely the mean of the spermidine values of postoperative patients with or without medical therapy and the spermine mean value for postoperative patients in tumor stage I and III were elevated. Cadaverine was never detected. The individual frequency of elevated polyamine levels in the plasma of single patients and variable, depending on patient subpopulation and polyamine under consideration. Spermidine was most frequently elevated (44%) in patients of tumor stage I under immunetherapy. We conclude that due to the low frequency of polyamine elevations to be found in plasma of melanoma patients by our method no diagnostic or prognostic significance and only limited importance for monitoring patients under therapy can be claimed for.

The therapeutic concept of irreversible inhibition of both ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) by alpha-difluoromethylornithine (DFMO) and methylglyoxal bis-guanylhydrazone (MGBG) is based on pathologic activities of these enzymes in tumor tissue. The polyamines putrescine, spermidine and spermine are measured in highest concentration in the prostate of both men and animals, with a significant increase of spermine in benign hyperplasia of the prostate. Patients with metastatic cancer of the prostate have elevated putrescine levels in the 24-hour urine. Treatment with 3 or 1% DFMO or 11 mg/kg MGBG in transplantable human and experimental cancer of the prostate demonstrated a significant anti-growth effect. A combination of DFMO and MGBG is tumor-destructive. The combination of 1% DFMO and 11 mg/kg MGBG distinctly reduces the activity of ODC and SAMDC and significantly lowers the levels of putrescine, spermidine and spermine in the tumor.