Assessment of MMP-9, TIMP-1, and COX-2 in normal tissue and in advanced symptomatic and asymptomatic carotid plaques

Abstract Background Mature carotid plaques are complex structures, and their histological classification is challenging. The carotid plaques of asymptomatic and symptomatic patients could exhibit identical histological components. Objectives To investigate whether matrix metalloproteinase 9 (MMP-9), tissue inhibitor of MMP (TIMP), and cyclooxygenase-2 (COX-2) have different expression levels in advanced symptomatic carotid plaques, asymptomatic carotid plaques, and normal tissue. Methods Thirty patients admitted for carotid endarterectomy were selected. Each patient was assigned preoperatively to one of two groups: group I consisted of symptomatic patients (n = 16, 12 males, mean age 66.7 ± 6.8 years), and group II consisted of asymptomatic patients (n = 14, 8 males, mean age 67.6 ± 6.81 years). Nine normal carotid arteries were used as control. Tissue specimens were analyzed for fibromuscular, lipid and calcium contents. The expressions of MMP-9, TIMP-1 and COX-2 in each plaque were quantified. Results Fifty-eight percent of all carotid plaques were classified as Type VI according to the American Heart Association Committee on Vascular Lesions. The control carotid arteries all were classified as Type III. The median percentage of fibromuscular tissue was significantly greater in group II compared to group I (p < 0.05). The median percentage of lipid tissue had a tendency to be greater in group I than in group II (p = 0.057). The percentages of calcification were similar among the two groups. MMP-9 protein expression levels were significantly higher in group II and in the control group when compared with group I (p < 0.001). TIMP-1 expression levels were significantly higher in the control group and in group II when compared to group I, with statistical difference between control group and group I (p = 0.010). COX-2 expression levels did not differ among groups. There was no statistical correlation between MMP-9, COX-2, and TIMP-1 levels and fibrous tissue. Conclusions MMP-9 and TIMP-1 are present in all stages of atherosclerotic plaque progression, from normal tissue to advanced lesions. When sections of a plaque are analyzed without preselection, MMP-9 concentration is higher in normal tissues and asymptomatic surgical specimens than in symptomatic specimens, and TIMP-1 concentration is higher in normal tissue than in symptomatic specimens.

TGF-β1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells

Abstract Background Metastasis is the main factor responsible for death in breast cancer patients. Matrix metalloproteinases (MMPs) and their inhibitors, known as tissue inhibitors of MMPs (TIMPs), and the membrane-associated MMP inhibitor (RECK), are essential for the metastatic process. We have previously shown a positive correlation between MMPs and their inhibitors expression during breast cancer progression; however, the molecular mechanisms underlying this coordinate regulation remain unknown. In this report, we investigated whether TGF-β1 could be a common regulator for MMPs, TIMPs and RECK in human breast cancer cell models. Methods The mRNA expression levels of TGF-β isoforms and their receptors were analyzed by qRT-PCR in a panel of five human breast cancer cell lines displaying different degrees of invasiveness and metastatic potential. The highly invasive MDA-MB-231 cell line was treated with different concentrations of recombinant TGF-β1 and also with pharmacological inhibitors of p38 MAPK and ERK1/2. The migratory and invasive potential of these treated cells were examined in vitro by transwell assays. Results In general, TGF-β2, TβRI and TβRII are over-expressed in more aggressive cells, except for TβRI, which was also highly expressed in ZR-75-1 cells. In addition, TGF-β1-treated MDA-MB-231 cells presented significantly increased mRNA expression of MMP-2, MMP-9, MMP-14, TIMP-2 and RECK. TGF-β1 also increased TIMP-2, MMP-2 and MMP-9 protein levels but downregulated RECK expression. Furthermore, we analyzed the involvement of p38 MAPK and ERK1/2, representing two well established Smad-independent pathways, in the proposed mechanism. Inhibition of p38MAPK blocked TGF-β1-increased mRNA expression of all MMPs and MMP inhibitors analyzed, and prevented TGF-β1 upregulation of TIMP-2 and MMP-2 proteins. Moreover, ERK1/2 inhibition increased RECK and prevented the TGF-β1 induction of pro-MMP-9 and TIMP-2 proteins. TGF-β1-enhanced migration and invasion capacities were blocked by p38MAPK, ERK1/2 and MMP inhibitors. Conclusion Altogether, our results support that TGF-β1 modulates the mRNA and protein levels of MMPs (MMP-2 and MMP-9) as much as their inhibitors (TIMP-2 and RECK). Therefore, this cytokine plays a crucial role in breast cancer progression by modulating key elements of ECM homeostasis control. Thus, although the complexity of this signaling network, TGF-β1 still remains a promising target for breast cancer treatment.

S-0 and S-1 State Structure, Methyl Torsional Barrier Heights, and Fast Intersystem Crossing Dynamics of 5-Methyl-2-hydroxypyrimidine

We report the analysis of the SI So rotational band contours of jet-cooled 5-methyl-2-hydroxypyrimidine (5M2HP), the enol form of deoxythymine. Unlike thymine, which exhibits a structureless spectrum, the vibronic spectrum of 5M2HP is well structured, allowing us to determine the rotational constants and the methyl group torsional barriers in the S-0 and S-1 states. The 0(0)(0), 6a(0)(1), 6b(0)(1), and 14(0)(1) band contours were measured at 900 MHz (0.03 cm(-1)) resolution using mass-specific two-color resonant two-photon ionization (2C-R2PI) spectroscopy. All four bands are polarized perpendicular to the pyrimidine plane (>90% c type), identifying the S-1 <- S-0 excitation of 5M2HP as a 1n pi* transition. All contours exhibit two methyl rotor subbands that arise from the lowest 5-methyl torsional states 0A '' and 1E ''. The S-0 and S-1 state torsional barriers were extracted from fits to the torsional subbands. The 3-fold barriers are V-3 '' = 13 cm(-1) and V3' = SI cm(-1); the 6-fold barrier contributions V-6 '' and V-6' are in the range of 2-3 cm(-1) and are positive in both states. The changes of A, B, and C rotational constants upon S-1 <- S-0 excitation were extracted from the contours and reflect an "anti-quinoidal" distortion. The 0(0)(0) contour can only be simulated if a 3 GHz Lorentzian line shape is included, which implies that the S-1(1n pi*) lifetime is similar to 55 ps. For the 6a(0)(1) and 6b(0)(1) bands, the Lorentzian component increases to 5.5 GHz, reflecting a lifetime decrease to similar to 30 ps. The short lifetimes are consistent with the absence of fluorescence from the 1n pi* state. Combining these measurements with the previous observation of efficient intersystem crossing (ISC) from the Si state to a long-lived T-1((3)n pi*) state that lies similar to 2200 cm(-1) below [S. Lobsiger, S. et al. Phys. Chem. Chem. Phys. 2010, 12, 5032] implies that the broadening arises from fast intersystem crossing with k(ISC) approximate to 2 x 10(10) s(-1). In comparison to 5-methylpyrimidine, the ISC rate is enhanced by at least 10 000 by the additional hydroxy group in position 2.

Filtration and oxidation characteristics of a diesel oxidation catalyst and a catalyzed particulate filter : development of a 1-D 2-layer model

The emissions, filtration and oxidation characteristics of a diesel oxidation catalyst (DOC) and a catalyzed particulate filter (CPF) in a Johnson Matthey catalyzed continuously regenerating trap (CCRT ®) were studied by using computational models. Experimental data needed to calibrate the models were obtained by characterization experiments with raw exhaust sampling from a Cummins ISM 2002 engine with variable geometry turbocharging (VGT) and programmed exhaust gas recirculation (EGR). The experiments were performed at 20, 40, 60 and 75% of full load (1120 Nm) at rated speed (2100 rpm), with and without the DOC upstream of the CPF. This was done to study the effect of temperature and CPF-inlet NO2 concentrations on particulate matter oxidation in the CCRT ®. A previously developed computational model was used to determine the kinetic parameters describing the oxidation characteristics of HCs, CO and NO in the DOC and the pressure drop across it. The model was calibrated at five temperatures in the range of 280 – 465° C, and exhaust volumetric flow rates of 0.447 – 0.843 act-m3/sec. The downstream HCs, CO and NO concentrations were predicted by the DOC model to within ±3 ppm. The HCs and CO oxidation kinetics in the temperature range of 280 - 465°C and an exhaust volumetric flow rate of 0.447 - 0.843 act-m3/sec can be represented by one ’apparent’ activation energy and pre-exponential factor. The NO oxidation kinetics in the same temperature and exhaust flow rate range can be represented by ’apparent’ activation energies and pre-exponential factors in two regimes. The DOC pressure drop was always predicted within 0.5 kPa by the model. The MTU 1-D 2-layer CPF model was enhanced in several ways to better model the performance of the CCRT ®. A model to simulate the oxidation of particulate inside the filter wall was developed. A particulate cake layer filtration model which describes particle filtration in terms of more fundamental parameters was developed and coupled to the wall oxidation model. To better model the particulate oxidation kinetics, a model to take into account the NO2 produced in the washcoat of the CPF was developed. The overall 1-D 2-layer model can be used to predict the pressure drop of the exhaust gas across the filter, the evolution of particulate mass inside the filter, the particulate mass oxidized, the filtration efficiency and the particle number distribution downstream of the CPF. The model was used to better understand the internal performance of the CCRT®, by determining the components of the total pressure drop across the filter, by classifying the total particulate matter in layer I, layer II, the filter wall, and by the means of oxidation i.e. by O2, NO2 entering the filter and by NO2 being produced in the filter. The CPF model was calibrated at four temperatures in the range of 280 – 465 °C, and exhaust volumetric flow rates of 0.447 – 0.843 act-m3/sec, in CPF-only and CCRT ® (DOC+CPF) configurations. The clean filter wall permeability was determined to be 2.00E-13 m2, which is in agreement with values in the literature for cordierite filters. The particulate packing density in the filter wall had values between 2.92 kg/m3 - 3.95 kg/m3 for all the loads. The mean pore size of the catalyst loaded filter wall was found to be 11.0 µm. The particulate cake packing densities and permeabilities, ranged from 131 kg/m3 - 134 kg/m3, and 0.42E-14 m2 and 2.00E-14 m2 respectively, and are in agreement with the Peclet number correlations in the literature. Particulate cake layer porosities determined from the particulate cake layer filtration model ranged between 0.841 and 0.814 and decreased with load, which is about 0.1 lower than experimental and more complex discrete particle simulations in the literature. The thickness of layer I was kept constant at 20 µm. The model kinetics in the CPF-only and CCRT ® configurations, showed that no ’catalyst effect’ with O2 was present. The kinetic parameters for the NO2-assisted oxidation of particulate in the CPF were determined from the simulation of transient temperature programmed oxidation data in the literature. It was determined that the thermal and NO2 kinetic parameters do not change with temperature, exhaust flow rate or NO2 concentrations. However, different kinetic parameters are used for particulate oxidation in the wall and on the wall. Model results showed that oxidation of particulate in the pores of the filter wall can cause disproportionate decreases in the filter pressure drop with respect to particulate mass. The wall oxidation model along with the particulate cake filtration model were developed to model the sudden and rapid decreases in pressure drop across the CPF. The particulate cake and wall filtration models result in higher particulate filtration efficiencies than with just the wall filtration model, with overall filtration efficiencies of 98-99% being predicted by the model. The pre-exponential factors for oxidation by NO2 did not change with temperature or NO2 concentrations because of the NO2 wall production model. In both CPF-only and CCRT ® configurations, the model showed NO2 and layer I to be the dominant means and dominant physical location of particulate oxidation respectively. However, at temperatures of 280 °C, NO2 is not a significant oxidizer of particulate matter, which is in agreement with studies in the literature. The model showed that 8.6 and 81.6% of the CPF-inlet particulate matter was oxidized after 5 hours at 20 and 75% load in CCRT® configuration. In CPF-only configuration at the same loads, the model showed that after 5 hours, 4.4 and 64.8% of the inlet particulate matter was oxidized. The increase in NO2 concentrations across the DOC contributes significantly to the oxidation of particulate in the CPF and is supplemented by the oxidation of NO to NO2 by the catalyst in the CPF, which increases the particulate oxidation rates. From the model, it was determined that the catalyst in the CPF modeslty increases the particulate oxidation rates in the range of 4.5 – 8.3% in the CCRT® configuration. Hence, the catalyst loading in the CPF of the CCRT® could possibly be reduced without significantly decreasing particulate oxidation rates leading to catalyst cost savings and better engine performance due to lower exhaust backpressures.

Ms. Ff. Mus. 1492 - Dom: XXIII. post: Trin: : Werthes Zion sey getrost pp ; â. C. A. T. B., violino primo, violino secundo, hautbois primo, hautbois secundo, clarino primo, clarino secundo, clarino tertio, tympani, viola 1, viola 2, violoncello, chalcedon e. organo

di Telemann

Na 1 Nachlass Max Horkheimer, 601 - Philosophische und ökomonische Notizen und Referate; Extracta I; Referat Transcendentale Systematik (p. VII 1, 1-14 - VII 3)

Hans Hildebrandt, "Der Kunstunterricht an Hochschulen", Typoskript, 2 Blatt; "Das Wirtschaftsprinzip", Typoskript, 3 Blatt (GS 11, S. 13-16); "Betr. Scheler - Sombart", Manuskript, 2 Blatt; "Zur Psychologie des Denkens nach Benno Erdmann", Manuskript, 2 Blatt, davon 1 leer; "Zur Phänomenologie der Erkenntnis", Typoskript, 3 Blatt; "Zu Kants Kritik der Urteilskraft", Typoskript mir eigenhändigen Korrekturen, 5 Blatt (GS 11, S. 70-75); "Naturwissenschaftliche Methode und Erkenntnistheorie. Über Kant", Typoskript mit zahlreichen eigenhändigen Notizen, 2 Blatt; "Beantwortung der Frage: Was heißt vom Standpunkt des Transcendentalen Idealismus aus die Behauptung, daß die materielle Entwicklung der Entwicklung des Bewußtseins vorhergegangen ist?", Typoskript, 3 Blatt (GS 11, S. 17-21); "Sprache und Denken", Typoskript, 8 Seiten (GS 11, S. 119-132); "Psychologie und Erkenntnistheorie", Typoskript, 2 Blatt; "Zu David Hume", Typoskript, 3 Blatt; "Wie ist das Bewußtsein einer Mehrheit von Gegenständen möglich?", Typoskript, 6 Blatt; "Gegenstand der Psychologie nach Cornelius", Typoskript, 2 Blatt; "Psychologie und Philosophie bei Hume und Cornelius", Typoskript, 12 Blatt; "Das Wiedererkennen", Typoskript von 19.12.1919, 8 Blatt; Exzerpte über: Gustav Landauer, Arthur Wolfgang Cohn, Karl Vorländer, Friedrich Engels, Karl Kautsky, L. B. Boudin, Karl Marx, Arthur Schopenhauer, 1 Heft, Manuskript, 42 Blatt; Manuskript, 1 Heft, 13 Blatt, davon 1 leer;

Na 1 Nachlass Max Horkheimer, 707 - "German Refugees from Eastern Zone" (p. IX 183.1-2 - IX 184.2-18)

"German Refugees from the Eastern Zone" 1952; 1. "Outline of a Pilot Study of German Refugees from the Eastern Zone Who are presently in Berlin" a) Typoskript, 4 Blatt; b) Typoskript, 3 Blatt; 2. "Budget for a Pilot Study of German Refugees from the Eastern Zone in Berlin" a) 2 Blatt; b) 1 Blatt; c) 1 Blatt; d) 1 Blatt; "Universität und Gesellschaft" 1952-1956; 1. "Universität und Gesellschaft" Teil III: Expertenbefragung. Forschungsbericht, 1953; als Typoskript vervielfältigt und gebunden, 77 Blatt; 2.-7. Allgemeine Darstellung der Untersuchungen; 2. "Einige wichtige Ergebnisse der Universitätsstudie", 1956. Als Typoskript vervielfältigt, 5 Blatt; 3. "Vorläufige Gliederung für den Bericht der Hochschul-Untersuchung" 2 Blatt; 4. "The German University of Today. Progress Report on a Research Project" 02.11.1955 a) Typoskirpt, 12 Blatt; b) Typoskript mit handschriftlichen Korrekturen, 12 Blatt; 5. "The German University of Today. A Progress Report on a Research Project" a) Typoskript, 7 Blatt; b) Typoskript mit handschriftlichen Korrekturen, 8 Blatt; c) Typoskirpt mit handschriftlichen Korrekturen, 12 Blatt; 6. "Funktion und Wirklichkeit der Universität heute. Zwischenbericht über drei Studien des Instituts für Sozialforschung an der Johann Wolfgang Goethe- Universität". Typoskript, 25 Blatt; 7. "Pläne einer Untersuchung über die Vorstellung von der Finktion der heutigen deutschen Universität bei bestimmten Personenkreisen" a) Typoskirpt, 11 Blatt; b) Typoskript, 11 Blatt; 8.-10. Studentenbefragung; 8. Fagebogen zur Umfrage "Warum studieren Studenten?", als Typoskript vervielfältigt, 19 Blatt; 9. "The Economic Situation of Students at the Johann Wolfgang Goethe- University in Frankfurt am Main" Resultate der Umfrage vom Winter 1952/43; Januar 1956; a) Typoskript, 43 Blatt; b) 43 Blatt; 10. "Students and Parentl Influence. Results of a Survey" a) Typoskript, 25 Blatt; b) Typoskript mit handschriftlichen Korrekturen von Frederick Pollock, 25 Blatt; 11.-14. Aktennotizen; 11. "Bericht über die Verhandlungen zwischen Herrn Professor Baumgarten, Tenbruck und Habermas am 12.12.1956 im Institut für vergleichende Sozialwissenschaften in Stuttgart. Betreff: Professorenstudie" 13.13.1956 vermutlich 13.03.1956. Typoskript, 2 Blatt; 12. Gembardt, Ulrich: Aktennotiz zur Hochschuluntersuchung. 23.05.1955, Typoskript, 3 Blatt; 13. Gembardt, Ulrich: Bemerkungen zur Aktennotiz vom 23.05.1955, Typoskript, 2 Blatt; 14. "Aktennotiz über die Hochschul-Forschungsprojekte des Göttinger Soziologischen Seminars und des Frankfurter Institut für Sozialforschung, die deim Hauptausschuß der Deutschen Forschungsgemeinschaft am 1. August 1953 zur Finanzierung vorgelegt werden" 20.07.1953. Typoskript, 2 Blatt; 15.-18. Briefe; 15. Löwenthal, Leo: 1 Brief mit Unterschrift an MAx Horkheimer, New York, 20.01.1955, 3 Blatt; 16. Horkheimer, Max [?]: 1 Brief an Chauncy D. Harris, ohne Ort, Januar 1955; a) Typoskirpt, 1 Blatt; b) Typoskirpt mit handschriftlichen Korrekturen, 1 Blatt; c) Typoskirpt mit handschriftlichen Korrekturen, 1 Blatt; d) Typoskirpt, 1 Blatt; 17. Horkheimer, Max: Drei gleichlautende Briefe an die Rektoren der Universität Bonn, Heidelberg und Koel, ohne Ort, 15.11.1953; a) Typoskript, 6 Blatt; b) Typoskript mit eigenhändigen KOrrekturen, 3 Blatt; 18. Plessner, Helmuth: 1 Brief mit Unterschrift mit Beilage an Max Horkheimer, Göttingen 02.07.1953, 4 Blatt;

Bcl-2 interacting protein, BAG-1, binds to and activates the kinase Raf-1.

The Bcl-2 protein blocks programmed cell death (apoptosis) through an unknown mechanism. Previously we identified a Bcl-2 interacting protein BAG-1 that enhances the anti-apoptotic effects of Bcl-2. Like BAG-1, the serine/threonine protein kinase Raf-1 also can functionally cooperate with Bcl-2 in suppressing apoptosis. Here we show that Raf-1 and BAG-1 specifically interact in vitro and in yeast two-hybrid assays. Raf-1 and BAG-1 can also be coimmunoprecipitated from mammalian cells and from insect cells infected with recombinant baculoviruses encoding these proteins. Furthermore, bacterially-produced BAG-1 protein can increase the kinase activity of Raf-1 in vitro. BAG-1 also activates this mammalian kinase in yeast. These observations suggest that the Bcl-2 binding protein BAG-1 joins Ras and 14-3-3 proteins as potential activators of the kinase Raf-1.

Role of neuronal nitric oxide in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurotoxicity.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) causes nigrostriatal dopaminergic pathway damage similar to that observed in Parkinson disease (PD). To study the role of NO radical in MPTP-induced neurotoxicity, we injected MPTP into mice in which nitric oxide synthase (NOS) was inhibited by 7-nitroindazole (7-NI) in a time- and dose-dependent fashion. 7-NI dramatically protected MPTP-injected mice against indices of severe injury to the nigrostriatal dopaminergic pathway, including reduction in striatal dopamine contents, decreases in numbers of nigral tyrosine hydroxylase-positive neurons, and numerous silver-stained degenerating nigral neurons. The resistance of 7-NI-injected mice to MPTP is not due to alterations in striatal pharmacokinetics or content of 1-methyl-4-phenylpyridinium ion (MPP+), the active metabolite of MPTP. To study specifically the role of neuronal NOS (nNOS), MPTP was administered to mutant mice lacking the nNOS gene. Mutant mice are significantly more resistant to MPTP-induced neurotoxicity compared with wild-type littermates. These results indicate that neuronally derived NO mediates, in part, MPTP-induced neurotoxicity. The similarity between the MPTP model and PD raises the possibility that NO may play a significant role in the etiology of PD.

Age determination of sediments of Site LO09-14 (Table 1)

A composite record (LO09-14) of three sediment cores from the subpolar North Atlantic (Reykjanes Ridge) was investigated in order to assess surface ocean variability during the last 11 kyr. The core site is today partly under the influence of the Irminger Current (IC), a branch of the North Atlantic Drift continuing northwestward around Iceland. However, it is also proximal to the Sub-Arctic Front (SAF) that may cause extra dynamic hydrographic conditions. We used statistical methods applied to the fossil assemblages of diatoms to reconstruct quantitative sea surface temperatures (SSTs). Our investigations give evidence for different regional signatures of Holocene surface oceanographic changes in the North Atlantic. Core LO09-14 reveal relatively low and highly variable SSTs during the early Holocene, indicating a weak IC and increased advection of subpolar water over the site. A mid-Holocene thermal optimum with a strong IC occurs from 7.5 to 5 kyr and is followed by cooler and more stable late Holocene surface conditions. Several intervals throughout the Holocene are dominated by the diatom species Rhizosolenia borealis, which we suggest indicates proximity to a strongly defined convergence front, most likely the SAF. Several coolings, reflecting southeastward advection of cold and ice-bearing waters, occur at 10.4, 9.8, 8.3, 7.9, 6.4, 4.7, 4.3 and 2.8 kyr. The cooling events recorded in the LO09-14 SSTs correlate well with both other surface records from the area and the NADW reductions observed at ODP Site 980 indicating a surface-deepwater linkage through the Holocene.