Synthese und Charakterisierung neuer funktionalisierter Mono- und Bis-tetrahydropyrrolo[3,4-b]carbazole als potentielle DNA-Liganden

Synthese und Charakterisierung neuer funktionalisierter Mono- und Bis-tetrahydro-pyrrolo[3,4-b]carbazole als potentielle DNA-Liganden In der Carbazol-Chemie sollen neue anellierte Verbindungen mit potentieller DNA-Affinität und damit verbundener Antitumoraktivität entwickelt werden. Auf molekularer Ebene sind DNA-Interkalation oder DNA-Rinnenbindung zu erwarten. Darauf aufbauend wurden in Anlehnung an literaturbekannte Cytostatika Mono- und Bis-tetrahydropyrrolo[3,4-b]carbazole synthetisiert, die zur Entwicklung neuer Leitstrukturen bzw. -substanzen beitragen können.In der vorliegenden Arbeit wurde als synthetische Schlüsselreaktion die in unserem Arbeitkreis etablierte Indol-2,3-chinodimethan-Diels-Alder-Reaktion mit geeigneten cyclischen Mono- und Bismaleinimiden als Dienophilen weiterführend genutzt. Auf Grund des Aufbaus von künftigen Struktur-Wirkungsbeziehungen wurden variable Linker zwischen die beiden zu verbindenden Pyrrolotetrahydrocarbazole eingeführt. Diese waren aliphatischer und diamidischer Natur. Diamidische Strukturelemente wurden im Hinblick auf die Entwicklung neuer Peptidomimetika eingeführt. Deren Synthese gelang zum einen über die gemischte Säureanhydrid-Methode und zum anderen über die Azolid-Methode. Die Struktursicherung der als Cycloaddukte erhaltenen Tetrahydrocarbazole erfolgte mittels Standardverfahren (1D-, 2D-NMR-, IR-Spektroskopie und Massenspektrometrie).Enantiomere bzw. Diastereomere chiraler Wirkstoffe unterscheiden sich stark in ihren pharmakologischen Eigenschaften, deshalb müssen Verfahren entwickelt werden, um diese Substanzen gegebenenfalls auch in enantiomerenreiner Form darstellen zu können. Die Racemate der Monotetrahydrocarbazole und die Racemate sowie die dazu diastereomeren meso-Formen der Bistetrahydrocarbazole, die bei der Reaktion entstehen, konnten erstmals mittels chiraler HPLC analytisch getrennt werden.In einer der Synthese ergänzten theoretischen Studie wurde Computer-Molecular-Modelling zur Problematik der Diels-Alder-Reaktion durchgeführt, außerdem wurden kraftfeld-mechanische Berechnungen zur Konformationsanalyse der 'einfachen' Monotetrahydro-carbazole herangezogen und darauf aufbauend schließlich einfache DNA-Docking-Experimente zur ersten Abschätzung des DNA-Binde-Verhaltens der synthetisierten Verbindungen vorgenommen.

Na 1 Nachlass Max Horkheimer, 690 - Materialien zu Forschungsprojekten über Antisemitismus u.a. (p. IX 157 - IX 163.1-9)

"Beitrag des Instituts für Sozialforschung zu dem Forschungsprojekt über Autorität" (3.1.1951). Typoskript, 5 Blatt; "Reactions to the Antisemitic Incidents in January 1960. A Pilot Study in Frankfurt am Main. Summary of Procedure and Results" (1960). Typoskript, 6 Blatt (= Alt.Sig. IX 234.13 a); "Proposal for International Study of Anti-Semitism" (1960):; 1. American Jewish Committee: Luncheon Meeting, May 25, 1960, Typoskript, 2 Blatt; 2. American Jewish Committee, Institute of Human Relations: "The JDA Agencies and Germany" (17.5.1960). Als Typoskript vervielfältigt, 32 Blatt; Exzerpte aus Werken über Antisemitismus, Literaturlisten (etwa 1933-46):; 1. Everett R. Clinchy, Typoskript, 1 Blatt; 2. Paul K. Hatt, Typoskript, 3 Blatt; 3. John Moffat Mecklin, Typoskript, 3 Blatt; 4. Conrad Henry Moehlman, Typoskript, 4 Blatt; 5. Maurice Samuel, Typoskript, 5 Blatt; 6. Milton Steinberg, Typoskript, 2 Blatt; 7. "General Literature on Antisemitism", Liste, Typoskript mit handschriftlichen Ergänzungen, 1 Blatt; 8. "Bibliography" zum Judentum, 15 Blatt; 9. Literaturliste, handschriftliche Notizen, 9 Blatt; 10. Literaturliste, eigenhändige Notizen von Max Horkheimer, 1 Blatt; 11. Literaturliste, 1 Blatt; 12. Zitate zum Judentum aus Zeitschriften, 1 Blatt; Memorandum zum Antisemitismus (1944-48):; 1. S. Andhil Fineberg: "Notes on 'A Mask for Privilege' by Carey McWilliams", als Typoskript vervielfältigt, 4 Blatt; Carey McWilliams: "Memorandum" (8.5.1948), Typoskript, 3 Blatt; Lawrence Bloomgarden und S.A. Fineberg: "In Reply to Carey McWilliams Memorandum of May 8th", Typoskript, 3 Blatt; 2. Rundbriefe der American Jewish Sociological Society, 1944, als Typoskript vervielfältigt, 4 Blatt; 3. Bericht über einen Vortrag von Wladimir Eliasberg über Antisemitismus, Typoskript, 1 Blatt; Abschriften und Übersetzungen aus Zeitungsartikeln über Antisemitismus (1939-43):; 1. "A Note on Anti-Semitism", aus: The New Statesman and Nation (13.3.1939), Typoskript, 7 Blatt; 2. Abschriften aus deutschen und englischen Zeitungen, 1939, Typoskript, 1 Blatt; 3. "A Homility of the Bishop of Cremona" (Übersetzung aus: Osservatore Romano) 1939. Typoskript, 18 Blatt; Veröffentlichungen über Antisemitismus, Vorurteil, Demagogie (1941-63):; 1. Earl Raab und Seymour M. Lipset: "Prejudice and Society", Freedom Pamphlet Anti-Defamation League of B'nai B'rith (New York 1963), 48 Seiten; 2. Committee on Education, Training and Research in Race Relations of the University of Chicago: Bulletin, Nr. 1, 30.6.1948, 55. Seiten; 3. Solomon Andhil Fineberg: "Checkmate for Rabble-Rousers. What to Do When the Demagogue Comes to Town", Sonderdruck aus Commentary, Vol. 2, 1946 und eine Broschüre, 20 Seiten; 4. Kurt Lewin: "A new Approach to Old Problems", aus: Congress Week, 19.1.1945, 2 Blatt; 5. Eric A. Johnston: "Intolerance", New York, 11.1.1945, Heft, 8 Blatt; 6. Philip Wylie: "Memorandum on Anti-Semitism", aus: American Mercury, Januar 1945, 5 Blatt; 7. S.I. Hayakawa: "Race and Words", aus: Common Sense, Juli 1943, 3 Blatt; 8. David Riesman: "The Politics of Persecution". Als Typoskript vervielfältigt, 21 Blatt; 9. James P. Gifford, Frank D. Schroth, Maximilian Moss, Edward A. Richards, Samuel J. Levinson, Thomas G. Grace: "Anti-Semitism. It's Causes and Cures", New York, 1941, 30 Seiten;

Amino acid concentrations in interstitial waters from ODP Sites 113-689 and 113-690

Biogenic calcareous and siliceous sediments were drilled at ODP Sites 689 and 690 on the Maud Rise, Antarctic Ocean. We analyzed dissolved combined amino acids (DCAA) and dissolved free amino acids (DFAA) in interstitial waters in order to characterize the amino acids in dissolved organic matter. The DFAA was predominant over the DCAA in interstitial waters at Sites 689 and 690, which contradicted the previous results from interstitial water and seawater studies. The DCAA in the interstitial waters probably originated from calcareous biogenic debris with less amounts of siliceous debris. Although glutamic acid constituted 41% of the total concentration of DCAA, it accounted for only 1% of the total concentration of DFAA due to the adsorption and/or reaction with biogenic carbonate. Ornithine, a nonprotein amino acid, is a decomposed product of arginine and made up 17 mol% of the total DFAA and. The total hydrolyzable amino acids (=DCAA + DFAA) accounted for 5 to 28% of the dissolved organic carbon (DOC) concentration, which implied that high molecular weight organic matter was a major contributor for the DOM (dissolved organic matter) in interstitial waters. Fairly positive correlation between the dissolved manganese and the total DCAA values suggested that the redox condition plays a significant role in controlling the total DCAA content. A small decrease in the sulfate concentration in the interstitial waters from both sites suggested fairly low microbial activity by sulfate-reducing bacteria.

Upper Cretaceous-Paleogene stratigraphy of ODP Sites 113-689 and 113-690

This contribution summarizes the biostratigraphy of planktonic foraminifers, calcareous nannofossils, and benthic foraminifers, in combination with the magnetostratigraphy, carbon and oxygen isotope stratigraphy of benthic foraminifers, and CaCO3 stratigraphy for the Maestrichtian through Paleogene calcareous sequences recovered at Sites 689 and 690 on Maud Rise (at about 65°S, eastern Weddell Sea, Antarctica). These data represent the southernmost calciumcarbonate record available for that interval, and thus extend the biostratigraphic and isotopic database to higher latitudes. Sites 689 and 690 form the southernmost anchor of a north-south transect through the Atlantic Ocean for Paleogene biostratigraphy and chemostratigraphy.

In vitro reconstitution of light-harvesting POR-protochlorophyllide complex with protochlorophyllides a and b

NADPH:protochlorophyllide oxidoreductase (POR; EC1.1.33.1) is a key enzyme for the light-induced greening of angiosperms. In barley, two POR proteins exist, termed PORA and PORB. These have previously been proposed to form higher molecular weight light-harvesting complexes in the prolamellar body of etioplasts (Reinbothe, C., Lebedev, N., and Reinbothe, S. (1999)Nature 397, 80–84). Here we report the in vitro reconstitution of such complexes from chemically synthesized protochlorophyllides (Pchlides) a andb and galacto- and sulfolipids. Low temperature (77 K) fluorescence measurements revealed that the reconstituted, lipid-containing complex displayed the same characteristics of photoactive Pchlide 650/657 as the presumed native complex in the prolamellar body. Moreover, Pchlide F650/657 was converted to chlorophyllide (Chlide) 684/690 upon illumination of the reconstituted complex with a 1-ms flash of white light. Identification and quantification of acetone-extractable pigments revealed that only the PORB-bound Pchlide a had been photoactive and was converted to Chlide a, whereas Pchlide b bound to the PORA remained photoinactive. Nondenaturing PAGE of the reconstituted Pchlide a/b-containing complex further demonstrated a size similar to that of the presumed native complexin vivo, suggesting that both complexes may be identical.

Action spectra for phytochrome A- and B-specific photoinduction of seed germination in Arabidopsis thaliana.

We have examined the seed germination in Arabidopsis thaliana of wild type (wt), and phytochrome A (PhyA)- and B (PhyB)-mutants in terms of incubation time and environmental light effects. Seed germination of the wt and PhyA-null mutant (phyA) was photoreversibly regulated by red and far-red lights of 10-1,000 micromol m-2 when incubated in darkness for 1-14 hr, but no germination occurred in PhyB-null mutant (phyB). When wt seeds and the phyB mutant seeds were incubated in darkness for 48 hr, they synthesized PhyA during dark incubation and germinated upon exposure to red light of 1-100 nmol m-2 and far-red light of 0.5-10 micromol m-2, whereas the phyA mutant showed no such response. The results indicate that the seed germination is regulated by PhyA and PhyB but not by other phytochromes, and the effects of PhyA and PhyB are separable in this assay. We determined action spectra separately for PhyA- and PhyB-specific induction of seed germination at Okazaki large spectrograph. Action spectra for the PhyA response show that monochromatic 300-780 nm lights of very low fluence induced the germination, and this induction was not photoreversible in the range examined. Action spectra for the PhyB response show that germination was photoreversibly regulated by alternate irradiations with light of 0.01-1 mmol m-2 at wavelengths of 540-690 nm and 695-780 nm. The present work clearly demonstrated that PhyA photoirreversibly triggers the germination upon irradiations with ultraviolet, visible and far-red light of very low fluence, while PhyB controls the photoreversible effects of low fluence.

Calcium carbonate concentrations of ODP Sites 113-689 and 113-690

An almost continuous Upper Cretaceous through Pleistocene biogenic sediment section was recovered from two sites on Maud Rise, a volcanic edifice in the Weddell Sea, off eastern Antarctica. Calcium carbonate values were determined for 1100 closely spaced samples using a Coulometrics CO2 Coulometer. Following a very brief decrease in the percentage of calcium carbonate immediately above the Cretaceous/Tertiary boundary, values remain high (~70%-80%), throughout most of the Paleocene, with variations primarily attributed to changes in the relative abundance of terrigenous and biogenic components. A small general decrease in calcium carbonate is observed from the upper Paleocene to lower middle Eocene. Eocene values continue to show small to moderate fluctuations. These fluctuations become more pronounced in the Oligocene as biosiliceous and carbonate sediments are mixed and interlayered. A distinct decrease in the calcium carbonate component is observed in the upper Oligocene through lower middle Miocene. Calcium carbonate becomes dominant again in the middle and lower upper Miocene, followed by almost exclusive biosiliceous sedimentation until the Pleistocene, where foraminifer-dominated calcareous ooze was recovered. Interpretation of this data will be carried out when a more finalized chronostratigraphy for the sequence has been produced.

An Asp49 Phospholipase A2 From Snake Venom Induces Cyclooxygenase-2 Expression And Prostaglandin E2 Production Via Activation Of Nf-κb, P38mapk, And Pkc In Macrophages.

Phospholipases A2 (PLA2) are key enzymes for production of lipid mediators. We previously demonstrated that a snake venom sPLA2 named MT-III leads to prostaglandin (PG)E2 biosynthesis in macrophages by inducing the expression of cyclooxygenase-2 (COX-2). Herein, we explored the molecular mechanisms and signaling pathways leading to these MT-III-induced effects. Results demonstrated that MT-III induced activation of the transcription factor NF-κB in isolated macrophages. By using NF-κB selective inhibitors, the involvement of this factor in MT-III-induced COX-2 expression and PGE2 production was demonstrated. Moreover, MT-III-induced COX-2 protein expression and PGE2 release were attenuated by pretreatment of macrophages with SB202190, and Ly294002, and H-7-dihydro compounds, indicating the involvement of p38MAPK, PI3K, and PKC pathways, respectively. Consistent with this, MT-III triggered early phosphorylation of p38MAPK, PI3K, and PKC. Furthermore, SB202190, H-7-dihydro, but not Ly294002 treatment, abrogated activation of NF-κB induced by MT-III. Altogether, these results show for the first time that the induction of COX-2 protein expression and PGE2 release, which occur via NF-κB activation induced by the sPLA2-MT-III in macrophages, are modulated by p38MAPK and PKC, but not by PI3K signaling proteins.

Staphylococcus Aureus Enterotoxins A And B Inhibit Human And Mice Eosinophil Chemotaxis And Adhesion In Vitro.

Staphylococcus aureus aggravates the allergic eosinophilic inflammation. We hypothesized that Staphylococcus aureus-derived enterotoxins directly affect eosinophil functions. Therefore, this study investigated the effects of Staphylococcal enterotoxins A and B (SEA and SEB) on human and mice eosinophil chemotaxis and adhesion in vitro, focusing on p38 MAPK phosphorylation and intracellular Ca(2+) mobilization. Eosinophil chemotaxis was evaluated using a microchemotaxis chamber, whereas adhesion was performed in VCAM-1 and ICAM-1-coated plates. Measurement of p38 MAPK phosphorylation and intracellular Ca(2+) levels were monitored by flow cytometry and fluorogenic calcium-binding dye, respectively. Prior incubation (30 to 240 min) of human blood eosinophils with SEA (0.5 to 3 ng/ml) significantly reduced eotaxin-, PAF- and RANTES-induced chemotaxis (P<0.05). Likewise, SEB (1 ng/ml, 30 min) significantly reduced eotaxin-induced human eosinophil chemotaxis (P<0.05). The reduction of eotaxin-induced human eosinophil chemotaxis by SEA and SEB was prevented by anti-MHC monoclonal antibody (1 μg/ml). In addition, SEA and SEB nearly suppressed the eotaxin-induced human eosinophil adhesion in ICAM-1- and VCAM-1-coated plates. SEA and SEB prevented the increases of p38 MAPK phosphorylation and Ca(2+) levels in eotaxin-activated human eosinophils. In separate protocols, we evaluated the effects of SEA on chemotaxis and adhesion of eosinophils obtained from mice bone marrow. SEA (10 ng/ml) significantly reduced the eotaxin-induced chemotaxis along with cell adhesion to both ICAM-1 and VCAM-1-coated plates (P<0.05). In conclusion, the inhibition by SEA and SEB of eosinophil functions (chemotaxis and adhesion) are associated with reductions of p38 MAPK phosphorylation and intracellular Ca(2+) mobilization.