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Calibration of TCCON column-averaged CO₂: the first aircraft campaign over European TCCON sites
(2011)
Messerschmidt, Janina ; Geibel, Marc Christoph ; Blumenstock, Thomas ; Chen, Hilin ; Deutscher, Nicholas M. ; Engel, Andreas ; Feist, Dietrich G. ; Gerbig, Christoph ; Gisi, Michael ; Hase, Frank ; Katrynski, Krzysztof ; Kolle, Olaf ; Lavrič, Jost-Valentin ; Notholt, Justus ; Palm, Mathias ; Ramonet, Michael ; Rettinger, Markus ; Schmidt, Martina ; Sussmann, Ralf ; Toon, Geoffrey C. ; Truong, Francois ; Warneke, Thorsten ; Wennberg, Paul O. ; Wunch, Debra ; Xueref-Remy, Irene
The Total Carbon Column Observing Network (TCCON) is a ground-based network of Fourier Transform Spectrometer (FTS) sites around the globe, where the column abundances of CO2, CH4, N2O, CO and O2 are measured. CO2 is constrained with a precision better than 0.25% (1-σ). To achieve a similarly high accuracy, calibration to World Meteorological Organization (WMO) standards is required. This paper introduces the first aircraft calibration campaign of five European TCCON sites and a mobile FTS instrument. A series of WMO standards in-situ profiles were obtained over European TCCON sites via aircraft and compared with retrievals of CO2 column amounts from the TCCON instruments. The results of the campaign show that the FTS measurements are consistently biased 1.1% ± 0.2% low with respect to WMO standards, in agreement with previous TCCON calibration campaigns. The standard a priori profile for the TCCON FTS retrievals is shown to not add a bias. The same calibration factor is generated using aircraft profiles as a priori and with the TCCON standard a priori. With a calibration to WMO standards, the highly precise TCCON CO2 measurements of total column concentrations provide a suitable database for the calibration and validation of nadir-viewing satellites
Calibration of TCCON column-averaged CO₂: the first aircraft campaign over European TCCON sites
(2011)
Messerschmidt, Janina ; Geibel, Marc Christoph ; Blumenstock, Thomas ; Chen, Hilin ; Deutscher, Nicholas M. ; Engel, Andreas ; Feist, Dietrich G. ; Gerbig, Christoph ; Gisi, Michael ; Hase, Frank ; Katrynski, Krzysztof Stanislaw ; Kolle, Olaf ; Lavrič, Jost-Valentin ; Notholt, Justus ; Palm, Mathias ; Ramonet, Michael ; Rettinger, Markus ; Schmidt, Martina ; Sussmann, Ralf ; Toon, Geoffrey C. ; Truong, Francois ; Warneke, Thorsten ; Wennberg, Paul O. ; Wunch, Debra ; Xueref-Remy, Irene
The Total Carbon Column Observing Network (TCCON) is a ground-based network of Fourier Transform Spectrometer (FTS) sites around the globe, where the column abundances of CO2, CH4, N2O, CO and O2 are measured. CO2 is constrained with a precision better than 0.25 %. To achieve a similarly high accuracy, calibration to World Meteorological Organization (WMO) standards is required. This paper introduces the first aircraft calibration campaign of five European TCCON sites and a mobile FTS instrument. A series of WMO standards in-situ profiles were obtained over European TCCON sites via aircraft and compared with retrievals of CO2 column amounts from the TCCON instruments. The results of the campaign show that the FTS measurements are consistently biased 1.0 % ± 0.2 % low with respect to WMO standards, in agreement with previous TCCON calibration campaigns. The standard a priori profile for the TCCON FTS retrievals is shown to not add a bias. The same calibration factor is generated using aircraft profiles as a priori and with the TCCON standard a priori. With a calibration to WMO standards, the highly precise TCCON CO2 measurements of total column concentrations provide a suitable database for the calibration and validation of nadir-viewing satellites.
Gharahi Ghehi, Nasrin ; Werner, Christian ; Cizungu Ntaboba, Landry ; Mbonigaba Muhinda, Jean Jacques ; Van Ranst, Eric ; Butterbach-Bahl, Klaus ; Kiese, Ralf ; Boeckx, Pascal
Globally, tropical forest soils represent the second largest source of N2O and NO. However, there is still considerable uncertainty on the spatial variability and soil properties controlling N trace gas emission. To investigate how soil properties affect N2O and NO emission, we carried out an incubation experiment with soils from 31 locations in the Nyungwe tropical mountain forest in southwestern Rwanda. All soils were incubated at three different moisture levels (50, 70 and 90% water filled pore space (WFPS)) at 17 °C. Nitrous oxide emission varied between 4.5 and 400 μg N m−2 h−1, while NO emission varied from 6.6 to 265 μg N m−2 h−1. Mean N2O emission at different moisture levels was 46.5 ± 11.1 (50% WFPS), 71.7 ± 11.5 (70% WFPS) and 98.8 ± 16.4 (90% WFPS) μg N m−2 h−1, while mean NO emission was 69.3 ± 9.3 (50% WFPS), 47.1 ± 5.8 (70% WFPS) and 36.1 ± 4.2 (90% WFPS) μg N m−2 h−1. The latter suggests that climate (i.e. dry vs. wet season) controls N2O and NO emissions. Positive correlations with soil carbon and nitrogen indicate a biological control over N2O and NO production. But interestingly N2O and NO emissions also showed a negative correlation (only N2O) with soil pH and a positive correlation with free iron. The latter suggest that chemo-denitrification might, at least for N2O, be an important production pathway. In conclusion improved understanding and process based modeling of N trace gas emission from tropical forests will not only benefit from better spatial explicit trace gas emission and basic soil property monitoring, but also by differentiating between biological and chemical pathways for N trace gas formation.
Die Labordiagnose einer Infektionskrankheit beruht auf dem Nachweis des Infektionserregers oder der spezifischen Immunreaktion unter Berücksichtigung der klinischen Plausibilität. Biologische Testverfahren wie der Zellkulturversuch erbringen nur näherungsweise ein quantitatives Ergebnis und sind mit einer relativ großen Streuung behaftet. Das gilt auch für Antikörperassays, soweit sie über ein biologisches Testsignal abgelesen werden (CPE, Agglutination, Komplementverbrauch). Moderne serologische und molekularbiologische Untersuchungsmethoden der Virologie werden i. d. R. über ein physikochemisches Testssignal abgelesen und quantitativ ausgewertet. Dadurch gelingt die nationale und internationale Standardisierung, die sich in Ringversuchen gut überprüfen lässt. Aus biologischen Gründen ist meist eine log. Ergebnisberechnung angezeigt, was für „ signifikante “ Unterschiede in Verlaufsuntersuchungen zu berücksichtigen ist: Da sowohl Infektion als auch Immunreaktion dynamische Prozesse darstellen, können Normalwerte in der virologischen Labordiagnostik nur restriktiv definiert werden. Ihre Ergebnisse sind mehr oder minder individuell interpretationsbedürftig.
Zum virologischen Nachweis einer akuten Influenza und zur Überprüfung des Immunstatus steht eine Vielzahl von Untersuchungsmethoden zur Verfügung. Bei Verdacht auf eine Influenzavirusinfektion liefert der Rachenabstrich das geeignete Untersuchungsmaterial. Das tiefe Nasopharynxaspirat ist etwas sensitiver, Sputum etwas weniger ergiebig. Die RT-PCR ermöglicht in 1–2 h nach Materialeingang ein sensitives und spezifisches Ergebnis. Typen, Subtypen und Driftvarianten lassen sich durch geeignete Primersonden, die kommerziell zur Verfügung stehen, einwandfrei identifizieren. Demgegenüber ist die Zellkultur-gestützte Virusisolierung zeitaufwendiger und stärker abhängig von einer sachgerechten Materialgewinnung und –überbringung (Kühlkette). PCR und Virusanzüchtung ermöglichen die geno- bzw. phänotypische Testung auf Therapieresistenzen. Der Antigentest ist eine einfache (bed-side) Schnellmethode. Seine Spezifität ist gut, die Sensitivität limitiert; daher kann der Antigentest nicht zur individuellen Ausschlussdiagnose eingesetzt werden. Influenzavirusspezifische Antikörper erscheinen im Blut erst in der zweiten Krankheitswoche. Die Serodiagnostik erfolgt typenspezifisch mit Komplementbindungsreaktion (KBR), IFT und ELISA über eine signifikante Titerbewegung oder den Nachweis von IgA-Antikörpern. IgG-spezifische IFT und ELISA Methoden geben Auskunft über die Influenzavirus-typspezifische Durchseuchung. Die klinisch relevantere subtypen- und variantenspezifische Influenzavirusimmunität wird mit dem HHT oder NT gemessen.
Adamczyk, Leszek ; Agakishiev, Geydar ; Aggarwal, Madan Mohan ; Ahammed, Zubayer ; Alakhverdyants, A. V. ; Alekseev, Igor G. ; Alford, J. ; Anderson, Bryon D. ; Anson, Christopher Daniel ; Arkhipkin, Dmitry A. ; Averichev, Georgy S. ; Balewski, Jan ; Banerjee, A. ; Barnovska, Z. ; Beavis, D. R. ; Bellwied, Rene ; Betancourt, M. J. ; Betts, R. R. ; Bhasin, Anju ; Bhati, Ashok Kumar ; Bichsel, Hans ; Bielčík, Jaroslav ; Bielčíková, Jana ; Bland, Leslie C. ; Bordyuzhin, Igor G. ; Borowski, W. ; Bouchet, J. ; Brandin, Andrei V. ; Brovko, S. G. ; Bruna, Elena ; Bueltmann, S. ; Bunzarov, Ivan ; Burton, T. P. ; Butterworth, Joseph ; Cai, Xiang-Zhou ; Caines, Helen Louise ; Calderón de la Barca Sánchez, Manuel ; Cebra, Daniel A. ; Cendejas, R. ; Cervantes, M. C. ; Chaloupka, Petr ; Chattopadhyay, S. ; Chen, H. F. ; Chen, J. H. ; Chen, J. Y. ; Chen, L. ; Cheng, J. ; Cherney, Michael Gerard ; Chikanian, Alexei ; Christie, William ; Chung, P. ; Chwastowski, J. ; Codrington, M. J. M. ; Corliss, R. ; Cramer, John G. ; Crawford, Henry J. ; Cui, X. ; Davila Leyva, A. ; De Silva, L. C. ; Debbe, R. R. ; Dedovich, T. G. ; Deng, J. ; Souza, Rafael Derradi de ; Dhamija, S. ; Didenko, Lidia ; Ding, F. ; Djawotho, P. ; Dong, X. ; Drachenberg, James Lucus ; Draper, James E. ; Du, C. M. ; Dunkelberger, L. E. ; Dunlop, James C. ; Efimov, Leonid G. ; Elnimr, M. ; Engelage, Jon M. ; Eppley, Geary Wareham ; Eun, L. ; Evdokimov, Olga Y. ; Fatemi, Renee H. ; Fedorišin, Jan ; Fersch, R. G. ; Filip, Peter ; Finch, Evan ; Fisyak, Yuri V. ; Gagliardi, Carl A. ; Gangadharan, Dhevan Raja ; Geurts, Frank ; Gliske, S. ; Gorbunov, Y. N. ; Grebenyuk, O. G. ; Grosnick, David P. ; Gupta, S. ; Guryn, Włodzimierz ; Haag, B. ; Hajkova, O. ; Hamed, Ahmed M. ; Han, L-X. ; Harris, John W. ; Hays-Wehle, J. P. ; Heppelmann, S. ; Hirsch, Andrew S. ; Hoffmann, Gerald W. ; Hofman, D. J. ; Horvat, S. ; Huang, Bingchu ; Huang, Huan Zhong ; Huck, P. ; Humanic, Thomas J. ; Huo, L. ; Igo, George J. ; Jacobs, William W. ; Jena, Chitrasen ; Joseph, J. ; Judd, Eleanor G. ; Kabana, Sonia ; Kang, K. ; Kapitan, J. ; Kauder, Kolja ; Ke, Hongwei ; Keane, Declan ; Kechechyan, Armen ; Kesich, A. ; Kettler, D. ; Kikola, D. P. ; Kiryluk, Joanna ; Kisiel, Adam Ryszard ; Kizka, V. ; Klein, Spencer Robert ; Koetke, Donald D. ; Kollegger, Thorsten ; Konzer, J. ; Koralt, I. ; Koroleva, L. ; Korsch, W. ; Kotchenda, L. ; Kravtsov, Peter ; Krueger, Keith William ; Kumar, Lokesh ; Lamont, Matthew A. C. ; Landgraf, Jeffery M. ; LaPointe, S. ; Lauret, Jerome ; Lebedev, Alexei ; Lednicky, Richard ; Lee, J. H. ; Leight, W. ; LeVine, Micheal J. ; Li, C. ; Li, L. ; Li, W. ; Li, X. ; Li, X. ; Li, Y. ; Li, Z. M. ; Lima, L. M. ; Lisa, Michael A. ; Liu, F. ; Ljubicic, Tonko A. ; Llope, William Joseph ; Longacre, Ronald S. ; Lu, Y. ; Luo, X. ; Luszczak, A. ; Ma, G. L. ; Ma, Yu-Gang ; Mahapatra, Durga Prasad ; Majka, Richard Daniel ; Mall, O. I. ; Margetis, Spyridon ; Markert, Christina ; Masui, H. ; Matis, Howard S. ; McDonald, Daniel ; McShane, Tom S. ; Mioduszewski, Saskia ; Mitrovski, M. K. ; Mohammed, Y. ; Mohanty, Bedangadas ; Morozov, B. ; Munhoz, Marcelo Gameiro ; Mustafa, M. K. ; Naglis, M. ; Nandi, Basanta Kumar ; Nasim, Md. ; Nayak, Tapan Kumar ; Nogach, Larisa V. ; Odyniec, Grazyna Janina ; Ogawa, Akio ; Oh, K. ; Ohlson, Alice ; Okorokov, Vitaly A. ; Oldag, E. W. ; Oliveira, R. A. N. ; Olson, Douglas L. ; Pachr, Miloš ; Page, Brian S. ; Pal, Susanta Kumar ; Pan, Y. X. ; Pandit, Y. ; Panebratsev, Yuri ; Pawlak, Tomasz Jan ; Pawlik, Bogdan ; Pei, Hua ; Perkins, Chris ; Peryt, Wiktor Stanislaw ; Pile, P. ; Planinic, Mirko ; Pluta, Jan Marian ; Plyku, D. ; Poljak, Nikola ; Porter, Jeffrey Brent ; Poskanzer, Arthur M. ; Powell, C. B. ; Prindle, Duncan J. ; Pruneau, Claude A. ; Pruthi, Navneet Kumar ; Przybycien, Mariusz ; Pujahari, P. R. ; Putschke, Jörn H. ; Qiu, H. ; Raniwala, Rashmi ; Raniwala, Sudhir ; Ray, Robert L. ; Redwine, R. ; Reed, Rosi Jan ; Riley, C. K. ; Ritter, Hans Georg ; Roberts, Jabus B. ; Rogačevskij, Oleg V. ; Romero, Juan L. ; Ruan, L. ; Rusnak, Jan ; Sahoo, Nihar Ranjan ; Sakrejda, Iwona ; Salur, Sevil ; Sandweiss, Jack H. ; Sangaline, E. ; Sarkar, Amal ; Schambach, Joachim ; Scharenberg, Rolf Paul ; Schmah, A. M. ; Schmitz, Norbert ; Schuster, T. R. ; Seele, J. ; Seger, Janet Elizabeth ; Seyboth, Peter ; Shah, N. ; Shahaliev, Ehtiram ; Shao, M. ; Sharma, B. ; Sharma, M. ; Shi, Shusu ; Shou, Q. Y. ; Sichtermann, Ernst P. ; Singaraju, Rama Narayana ; Skoby, Michael J. ; Smirnov, Nikolai ; Solanki, D. ; Sorensen, Paul Richard ; deSouza, U. G. ; Spinka, Harold. M. ; Srivastava, B. ; Stanislaus, T. D. Shirvel ; Steadman, S. G. ; Stevens, J. R. ; Stock, Reinhard ; Strichanov, Michail Nikolaevič ; Stringfellow, Blair ; Suaide, Alexandre Alarcon do Passo ; Suarez, M. C. ; Šumbera, Michal ; Sun, X. M. ; Sun, Y. ; Sun, Z. ; Surrow, Bernd ; Svirida, Dmitry N. ; Symons, Timothy James MacNeil ; Toledo, Alejandro Szanto de ; Takahashi, Jun ; Tang, Aihong ; Tang, Zebo ; Tarini, L. H. ; Tarnowsky, Terence J. ; Thein, Dylan ; Thomas, James H. ; Tian, J. ; Timmins, Anthony Robert ; Tlusty, David ; Tokarev, Mikhail V. ; Trainor, Thomas A. ; Trentalange, Stephen ; Tribble, Robert E. ; Tribedy, Prithwish ; Trzeciak, Barbara Antonina ; Tsai, Oleg D. ; Turnau, J. ; Ullrich, Thomas ; Underwood, David G. ; Van Buren, Gene ; van Nieuwenhuizen, G. ; Vanfossen, J. A. ; Varma, Raghava ; Vasconcelos, G. M. S. ; Videbæk, Flemming ; Viyogi, Yogendra P. ; Vokal, Stanislav ; Voloshin, Sergei ; Vossen, A. ; Wada, M. ; Wang, F. ; Wang, G. ; Wang, H. ; Wang, J. S. ; Wang, Q. ; Wang, X. L. ; Wang, Y. ; Webb, Grant ; Webb, Jason C. ; Westfall, Gary D. ; Whitten, Charles A. ; Wieman, Howard ; Wissink, Scott W. ; Witt, Richard ; Witzke, W. ; Wu, Y. F. ; Xiao, Z. ; Xie, W. ; Xin, K. ; Xu, H. ; Xu, N. ; Xu, Qinghua ; Xu, W. ; Xu, Y. ; Xu, Z. ; Xue, L. ; Yang, Y. ; Yang, Y. ; Yepes, Pablo P. ; Yi, Y. ; Yip, K. ; Yoo, I-K. ; Zawisza, M. ; Zbroszczyk, Hanna Paulina ; Zhang, J. B. ; Zhang, S. ; Zhang, W. M. ; Zhang, X. P. ; Zhang, Y. ; Zhang, Z. P. ; Zhao, F. ; Zhao, J. ; Zhong, C. ; Zhu, X. ; Zhu, Y. H. ; Zoulkarneeva, Youlia
STAR's measurements of directed flow (v1) around midrapidity for π±, K±, K0S, p and p¯ in Au + Au collisions at $\sqrtsNN = 200$ GeV are presented. A negative v1(y) slope is observed for most of produced particles (π±, K±, K0S and p¯). The proton v1(y) slope is found to be much closer to zero compared to antiprotons. A sizable difference is seen between v1 of protons and antiprotons in 5-30% central collisions. The v1 excitation function is presented. Comparisons to model calculations (RQMD, UrQMD, AMPT, QGSM with parton recombination, and a hydrodynamics model with a tilted source) are made. Anti-flow alone cannot explain the centrality dependence of the difference between the v1(y) slopes of protons and antiprotons.
Blaess, Sandra ; Bodea, Gabriela O. ; Kabanova, Anna ; Chanet, Soline ; Mugniery, Emilie ; Derouiche, Amin ; Stephen, Daniel ; Joyner, Alexandra L.
Background: The ventral midbrain contains a diverse array of neurons, including dopaminergic neurons of the ventral tegmental area (VTA) and substantia nigra (SN) and neurons of the red nucleus (RN). Dopaminergic and RN neurons have been shown to arise from ventral mesencephalic precursors that express Sonic Hedgehog (Shh). However, Shh expression, which is initially confined to the mesencephalic ventral midline, expands laterally and is then downregulated in the ventral midline. In contrast, expression of the Hedgehog target gene Gli1 initiates in the ventral midline prior to Shh expression, but after the onset of Shh expression it is expressed in precursors lateral to Shh-positive cells. Given these dynamic gene expression patterns, Shh and Gli1 expression could delineate different progenitor populations at distinct embryonic time points. Results: We employed genetic inducible fate mapping (GIFM) to investigate whether precursors that express Shh (Shh-GIFM) or transduce Shh signaling (Gli1-GIFM) at different time points give rise to different ventral midbrain cell types. We find that precursors restricted to the ventral midline are labeled at embryonic day (E)7.5 with Gli1-GIFM, and with Shh-GIFM at E8.5. These precursors give rise to all subtypes of midbrain dopaminergic neurons and the anterior RN. A broader domain of progenitors that includes the ventral midline is marked with Gli1-GIFM at E8.5 and with Shh-GIFM at E9.5; these fate-mapped cells also contribute to all midbrain dopaminergic subtypes and to the entire RN. In contrast, a lateral progenitor domain that is labeled with Gli1-GIFM at E9.5 and with Shh-GIFM at E11.5 has a markedly reduced potential to give rise to the RN and to SN dopaminergic neurons, and preferentially gives rise to the ventral-medial VTA. In addition, cells derived from Shh- and Gli1-expressing progenitors located outside of the ventral midline give rise to astrocytes. Conclusions: We define a ventral midbrain precursor map based on the timing of Gli1 and Shh expression, and suggest that the diversity of midbrain dopaminergic neurons is at least partially determined during their precursor stage when their medial-lateral position, differential gene expression and the time when they leave the ventricular zone influence their fate decisions.
Haberlandt, Christian ; Derouiche, Amin ; Wyczynski, Alexandra ; Haseleu, Julia ; Pohle, Jörg ; Karram, Khalad ; Trotter, Jacqueline ; Seifert, Gerald ; Frotscher, Michael ; Steinhäuser, Christian ; Jabs, Ronald
NG2 cells, the fourth type of glia in the mammalian CNS, receive synaptic input from neurons. The function of this innervation is unknown yet. Postsynaptic changes in intracellular Ca2+-concentration ([Ca2+]i) might be a possible consequence. We employed transgenic mice with fluorescently labeled NG2 cells to address this issue. To identify Ca2+-signaling pathways we combined patch-clamp recordings, Ca2+-imaging, mRNA-transcript analysis and focal pressure-application of various substances to identified NG2-cells in acute hippocampal slices. We show that activation of voltage-gated Ca2+-channels, Ca2+-permeable AMPA-receptors, and group I metabotropic glutamate-receptors provoke [Ca2+]i-elevations in NG2 cells. The Ca2+-influx is amplified by Ca2+-induced Ca2+-release. Minimal electrical stimulation of presynaptic neurons caused postsynaptic currents but no somatic [Ca2+]i elevations, suggesting that [Ca2+]i elevations in NG2 cells might be restricted to their processes. Local Ca2+-signaling might provoke transmitter release or changes in cell motility. To identify structural prerequisites for such a scenario, we used electron microscopy, immunostaining, mRNA-transcript analysis, and time lapse imaging. We found that NG2 cells form symmetric and asymmetric synapses with presynaptic neurons and show immunoreactivity for vesicular glutamate transporter 1. The processes are actin-based, contain ezrin but not glial filaments, microtubules or endoplasmic reticulum. Furthermore, we demonstrate that NG2 cell processes in situ are highly motile. Our findings demonstrate that gray matter NG2 cells are endowed with the cellular machinery for two-way communication with neighboring cells.
Martin-Subero, Jose Ignacio ; Ammerpohl, Ole ; Bibikova, Marina ; Wickham-Garcia, Eliza ; Agirre, Xabier ; Alvarez, Sara ; Brüggemann, Monika ; Bug, Stefanie ; Calasanz, Maria J. ; Deckert, Martina ; Dreyling, Martin ; Du, Ming Q. ; Dürig, Jan ; Dyer, Martin J. S. ; Fan, Jian-Bing ; Gesk, Stefan ; Hansmann, Martin-Leo ; Harder, Lana ; Hartmann, Sylvia ; Klapper, Wolfram ; Küppers, Ralf ; Montesinos-Rongen, Manuel ; Nagel, Inga ; Pott, Christiane ; Richter, Julia ; Román-Gómez, José ; Seifert, Marc ; Stein, Harald ; Suela, Javier ; Trümper, Lorenz ; Vater, Inga ; Prosper, Felipe ; Haferlach, Claudia ; Cruz Cigudosa, Juan ; Siebert, Reiner
Background: Alterations in the DNA methylation pattern are a hallmark of leukemias and lymphomas. However, most epigenetic studies in hematologic neoplasms (HNs) have focused either on the analysis of few candidate genes or many genes and few HN entities, and comprehensive studies are required. Methodology/Principal Findings: Here, we report for the first time a microarray-based DNA methylation study of 767 genes in 367 HNs diagnosed with 16 of the most representative B-cell (n = 203), T-cell (n = 30), and myeloid (n = 134) neoplasias, as well as 37 samples from different cell types of the hematopoietic system. Using appropriate controls of B-, T-, or myeloid cellular origin, we identified a total of 220 genes hypermethylated in at least one HN entity. In general, promoter hypermethylation was more frequent in lymphoid malignancies than in myeloid malignancies, being germinal center mature B-cell lymphomas as well as B and T precursor lymphoid neoplasias those entities with highest frequency of gene-associated DNA hypermethylation. We also observed a significant correlation between the number of hypermethylated and hypomethylated genes in several mature B-cell neoplasias, but not in precursor B- and T-cell leukemias. Most of the genes becoming hypermethylated contained promoters with high CpG content, and a significant fraction of them are targets of the polycomb repressor complex. Interestingly, T-cell prolymphocytic leukemias show low levels of DNA hypermethylation and a comparatively large number of hypomethylated genes, many of them showing an increased gene expression. Conclusions/Significance: We have characterized the DNA methylation profile of a wide range of different HNs entities. As well as identifying genes showing aberrant DNA methylation in certain HN subtypes, we also detected six genes—DBC1, DIO3, FZD9, HS3ST2, MOS, and MYOD1—that were significantly hypermethylated in B-cell, T-cell, and myeloid malignancies. These might therefore play an important role in the development of different HNs.
Background: Parkinson's disease (PD) is a slowly progressive neurodegenerative disorder which affects widespread areas of the brainstem, basal ganglia and cerebral cortex. A number of proteins are known to accumulate in parkinsonian brains including ubiquitin and alpha-synuclein. Prion diseases are sporadic, genetic or infectious disorders with various clinical and histopathological features caused by prion proteins as infectious proteinaceous particles transmitting a misfolded protein configuration through brain tissue. The most important form is Creutzfeldt-Jakob disease which is associated with a self-propagating pathological precursor form of the prion protein that is physiologically widely distributed in the central nervous system. Discussion: It has recently been found that alpha-synuclein may behave similarly to the prion precursor and propagate between cells. The post-mortem proof of alpha-synuclein containing Lewy bodies in embryonic dopamine cells transplants in PD patient suggests that the misfolded protein might be transmitted from the diseased host to donor neurons reminiscent of prion behavior. The involvement of the basal ganglia and brainstem in the degenerative process are other congruencies between Parkinson's and Creutzfeldt-Jakob disease. However, a number of issues advise caution before categorizing Parkinson's disease as a prion disorder, because clinical appearance, brain imaging, cerebrospinal fluid and neuropathological findings exhibit fundamental differences between both disease entities. Most of all, infectiousness, a crucial hallmark of prion diseases, has never been observed in PD so far. Moreover, the cellular propagation of the prion protein has not been clearly defined and it is, therefore, difficult to assess the molecular similarities between the two disease entities. Summary: At the current state of knowledge, the molecular pathways of transmissible pathogenic proteins are not yet fully understood. Their exact involvement in the pathophysiology of prion disorders and neurodegenerative diseases has to be further investigated in order to elucidate a possible overlap between both disease categories that are currently regarded as distinct entities.
According to the World Health Organization (WHO) bacterial resistance to antibiotic drug therapy is emerging as a major public health problem around the world. Infectious diseases seriously threaten the health and economy of all countries. Hence, the preservation of the effectiveness of antibiotics is a world wide priority. The key to preserving the power of antibiotics lies in maintaining their diversity. Many microorganisms are capable of producing these bioactive products, the so called antibiotics. Specifically in microorganisms, polyketide synthases (PKS) and non-ribosomal peptide synthases (NRPS) produce these natural bioactive compounds. Besides being used as antibiotics these non-ribosomal peptides and polyketides display an even broader spectrum of biological activities, e.g. as antivirals, immunosuppressants or in antitumor therapy. The wide functional spectrum of the peptides and ketides is due to their structural diversity. Mostly they are cyclic or branched cyclic compounds, containing non-proteinogenic amino acids, small heterocyclic rings and other unusual modifications such as epimerization, methylation, N‐formylation or heterocyclization. It is has been shown that these modifications are important for biological activity, but little is known about their biosynthetic origin.
PKS and NRPS are multidomain protein assembly lines which function by sequentially elongating a growing polyketide or peptide chain by incorporating acyl units or amino acids, respectively. The growing product is attached via a thioester linkage to the 4’-phosphopantetheine (4’-Ppant) arm of a holo acyl carrier protein (ACP) in PKSs or holo peptidyl carrier protein (PCP) in NRPSs and is passed from one module to another along the chain of reaction centers. The modular arrangement makes PKS and NRPS systems an interesting target for protein engineering. More than 200 novel polyketide compounds have already been created by module swapping, gene deletion or other specific manipulations. Unfortunately, however, engineered PKS often fail to produce significant amounts of the desired products. Structural studies may faciliate yield improvement from engineered systems by providing a more complete understanding of the interface between the different domains. While some information about domain-domain interactions, involving the most common enzymatic modules, ketosynthase and acyltransferase, is starting to emerge, little is known about the interaction of ACP domains with other modifying enzymes such as methyltransferases, epimerases or halogenases.
To further improve the understanding of domain-domain interactions this work focuses on the curacin A assembly line. Curacin A, which exhibits anti-mitotic activity, is from the marine cyanobacterium Lyngbya majuscula. This outstanding natural product contains a cyclopropane ring, a thiazoline ring, an internal cis double bond and a terminal alkene. The biosynthesis of curacin A is performed by a 2.2 Mega Dalton (MDa) hybrid PKS-NRPS cluster. A 10-enzyme assembly catalyzes the formation of the cyclopropane moiety as the first building block of the final product. Interestingly, for these enzymes the substrate is presented by an unusual cluster of three consecutive ACPs (ACPI,II,III). Little is known about the function of multiple ACPs which are supposed to increase the overall flux for enhanced production of secondary metabolites.
The first task in this work was to elucidate the structural effect of the triplet ACP repetition by nuclear magnetic resonance (NMR). The initial data show that the excised ACPI, ACPII or ACPIII proteins resulted in [15N, 1H]-TROSY spectra with strong chemical shift perturbations (CSPs), suggesting an effect on the structure. The triplet ACP domains display a high sequence identity (93- 100%) making structural investigation using usual NMR techniques due to high peak overlap impossible. To enable the investigation of the triplet ACP in its native composition we developed a powerful method, the three fragment ligation. Segmental labeling allows incorporating isotopes into one single domain in its multidomain context. As a result we could prepare the triplet ACP with only one domain isotopically labeled and therefore assign the full length protein. In this way our method paved the way to study the structural effects of the triplet ACP repetition. We could show unexpectedly, that, despite the fact that the triplet repeat of CurA ACPI,II,III has a synergistic effect in the biosynthesis of CurA, the domains are structurally independent.
In the second part of this work, we studied the structure of the isolated ACPI domain. Our results show that the CurA ACPI undergoes no major conformational changes upon activation via phosphopantetheinylation and therefore contradicts the conformational switching model which has been proposed for PCPs. Further we report the NMR solution structures of holo-ACPI and 3-hydroxyl-3-methylglutaryl (HMG)-ACPI. Data obtained from filtered nuclear overhauser effect (NOE) experiments indicate that the substrate HMG is not sequestered but presented on the ACP surface.
In the third part of this work we focussed on the protein-protein interactions of the isolated ACPI with its cognate interaction partners. We were especially interested in the interaction with the halogenase (Cur Hal), the first enzyme within the curacin A sub-cluster, acting on the initial hydroxyl-methyl-glutaryl (HMG) attached to ACPI. Primarily we studied the interaction using NMR titration and fluorescence anisotropy measurements. Surprisingly no complex between ACPI and Cur Hal could be detected. The combination of an activity assay using matrix-assisted laser desorption/ionization (MALDI) mass spectroscopy and mutational analysis revealed several amino acids of ACPI that strongly decrease the activity of CurA Hal. Mapping these mutations according to their effect on the Cur Hal activity onto the structure of HMG-ACPI displays that these amino acids surround the substrate and form a consecutive surface. These results suggest that this surface is important for Cur Hal recognition and selectivity. Our research presented herein is an excellent example for protein-protein interactions in PKS systems underlying a specific recognition process.
VHF ist mit einer erhöhten Morbidität und Mortalität assoziiert. Aufgrund des erhöhten thromboembolischen Risikos erfolgt eine Risikostratifizierung im klinischen Alltag mit Hilfe des CHADS2 Scores. Dieser ist praktikabel, jedoch oftmals ungenau, da nicht alle Risikofaktoren erfasst werden. Klinische und experimentelle Daten deuten auf einen kausalen Zusammenhang zwischen systemischer Immunreaktion und Entstehung von VHF als auch von VHF-abhängigen Komplikationen. In mehreren Studien zeigte sich, dass CRP ein prognostischer Risikofaktor für zukünftige kardiovaskuläre Erkrankungen ist. Darüber hinaus spielen auch andere Enzyme (wie z.B. MMPs) eine Rolle bei der Modulation von Entzündungsreaktionen im Rahmen kardiovaskulärer Erkrankungen.
Ziel dieser Studie: Identifizierung von Serum-Markern als Prädiktoren für zukünftige, thromboembolische Komplikationen bei Patienten mit VHF zur optimierten Risikostratifizierung hinsichtlich kardiovaskulärer Ereignisse. Hierzu wurden folgende Biomarker untersucht: MMP2, sVCAM1, hsCRP, vWF und sCD40L.
Material und Methode: Zur Analyse der prädiktiven Wertigkeit biochemischer Marker für das Auftreten thromboembolischer Komplikationen bei VHF wurden Patienten über einen langen Zeitraum nachbeobachtet. Es wurde das Auftreten des kombinierten, primären Endpunktes (Herzinfarkt, Schlaganfall, periphere Embolie, TIA, Tod) und des sekundären Endpunktes (Gesamtmortalität) untersucht. Die Serum-Marker Konzentrationen wurden mittels ELISA-Technik aus EDTA Blut bestimmt.
Ergebnisse: Es wurden 278 Patienten in die Studie eingeschlossen. Hiervon waren 175 (63%) männlich. Das mittlere Alter betrug 70,3 Jahre (37,5 bis 94,5 Jahre); Insgesamt wurden 272 Patienten über im Mittel 27±1 Monate nachbeobachtet. Paroxysmales VHF lag bei 198 (71%) Patienten vor, 39 (14%) hatten persistierendes und 41 (15%) permanentes VHF. Arterielle Hypertonie: 214 (77%) Patienten; KHK: 113 (41%) Patienten; 73 (26%) Patienten hatten Diabetes mellitus, 40 (14%) Patienten hatten vor Studieneinschluss einen Schlaganfall oder eine TIA. Bei 22 (8%) Patienten wurde eine Kreatinin-Clearance < 30%, bei 50 (18%) Patienten eine Mitralinsuffizienz > II° sowie bei 5 (2%) Patienten eine Aortenstenose > II° dokumentiert. Die LVEF betrug durchschnittlich 48±18%. LVEF <35%: 89 (32%) Patienten; 88 (32,4%) Patienten erreichten ein dem primären Endpunkt zugeordnetes Ereignis, davon erlitten 3 (1,1%) Personen einen Herzinfarkt, 8 (2,9%) Personen einen Schlaganfall oder eine TIA und 2 Personen (0,7%) eine periphere Embolie. Insgesamt verstarben 75 (27,6%) Patienten (nach durchschnittlich 15 Monaten) und erreichten somit den sekundären Endpunkt. In der univariaten Analyse waren in Bezug auf den primären und sekundären Endpunkt Alter > 75 Jahre, Diabetes mellitus, LVEF <35%, CHADS2 Score >2, hohe MMP2-, vWF- und hohe sVCAM-Werte signifikant assoziiert. In der multivariaten Analyse zeigte sich, dass hohes Alter, hohe MMP2- und sVCAM1-Werte unabhängig und signifikant mit dem primären Endpunkt assoziiert waren, während mit dem sekundären Endpunkt zusätzlich LVEF <35% signifikant assoziiert war.
Diskussion: Neben fortgeschrittenem Alter korrelierten erhöhte sVCAM1 und MMP2 Werte mit dem Auftreten kardiovaskulärer Ereignisse (Myokardinfarkt, Schlaganfall/TIA, Tod) sowie mit einem erhöhten Mortalitätsrisiko. VHF korrelierte in früheren Arbeiten mit erhöhten Entzündungsmarkern und mit VHF-Komplikationen. In unserer Arbeit konnte kein unabhängiger Einfluss von hsCRP auf den primären oder sekundären Endpunkt nachgewiesen werden. Möglicherweise ist eine Ursache darin zu suchen, dass die verschiedenen Marker von Inflammation, Gewebeumbau und Zelladhäsion bisher noch nicht systematisch untersucht wurden.
Endotheliales Remodeling der Vorhöfe und der Umbau der extrazellulären Matrix kann für die Entstehung eines Schlaganfalls von Bedeutung sein kann und korreliert mit erhöhten MMP2 Werten. Unsere Daten zeigen zusammenfassend, dass Marker struktureller oder endothelialer Umbauvorgänge bei VHF Patienten stärker mit kardiovaskulären Ereignissen assoziiert sind als hsCRP.
Girmatsion, Zenawit ; Biliczki, Peter ; Takac, Ina ; Schwerthelm, Christin ; Hohnloser, Stefan H. ; Ehrlich, Joachim
Background and Objective: The slow delayed rectifier current (IKs) is important for cardiac action potential termination. The underlying channel is composed of Kv7.1 α-subunits and KCNE1 β-subunits. While most evidence suggests a role of KCNE1 transmembrane domain and C-terminus for the interaction, the N-terminal KCNE1 polymorphism 38G is associated with reduced IKs and atrial fibrillation (a human arrhythmia). Structure-function relationship of the KCNE1 N-terminus for IKs modulation is poorly understood and was subject of this study.
Methods: We studied N-terminal KCNE1 constructs disrupting structurally important positively charged amino-acids (arginines) at positions 32, 33, 36 as well as KCNE1 constructs that modify position 38 including an N-terminal truncation mutation. Experimental procedures included molecular cloning, patch-clamp recording, protein biochemistry, real-time-PCR and confocal microscopy.
Results: All KCNE1 constructs physically interacted with Kv7.1. IKs resulting from co-expression of Kv7.1 with non-atrial fibrillation ‘38S’ was greater than with any other construct. Ionic currents resulting from co-transfection of a KCNE1 mutant with arginine substitutions (‘38G-3xA’) were comparable to currents evoked from cells transfected with an N-terminally truncated KCNE1-construct (‘Δ1-38’). Western-blots from plasma-membrane preparations and confocal images consistently showed a greater amount of Kv7.1 protein at the plasma-membrane in cells co-transfected with the non-atrial fibrillation KCNE1-38S than with any other construct.
Conclusions: The results of our study indicate that N-terminal arginines in positions 32, 33, 36 of KCNE1 are important for reconstitution of IKs. Furthermore, our results hint towards a role of these N-terminal amino-acids in membrane representation of the delayed rectifier channel complex.
Das letzte neue Medium, dem man – egal ob Gegner oder Freund – zubilligen muss, dass es unsere Welt fundamental verändert (hat), ist das Internet. Technische und historische Entwicklungen erspare ich mir an dieser Stelle und verweise auf die entsprechenden Darstellungen im – na? – im Internet. Wenn man dumme oder sagen wir vielleicht lieber peinliche Zitate übers Internet bzw. die daraus resultierenden gesellschaftlichen Debatten sucht, dann stößt man auf wirklich überraschende Stilblüten. So entblödete sich Stephan Holthoff-Pförtner, Gesellschafter der WAZ, nicht, Bloggern den Schutz des Artikels 5 GG abzusprechen. Angesichts der heutigen Bedeutung sozialer Netzwerke im Alltag lag auch BILD-Kolumnist Franz Josef Wagner im Jahr 2006 falsch, als er erklärte: "Einem Menschen wird man auf seinem Weg zum Bäcker begegnen, aber niemals im Internet." ...
Bingemer, Heinz ; Klein, Holger ; Ebert, Martin ; Haunold, Werner ; Bundke, Ulrich ; Herrmann, Thomas ; Kandler, Konrad ; Müller-Ebert, Dörthe ; Weinbruch, Stephan ; Judt, Anna ; Ardon-Dryer, Karin ; Levin, Zev ; Curtius, Joachim
Explosive volcanism affects weather and climate. Primary volcanic ash particles which act as ice nuclei (IN) can modify the phase and properties of cold tropospheric clouds. During the Eyjafjallajökull volcanic eruption we have measured the highest ice nucleus number concentrations (>600 L) in our record of 2 years of daily IN measurements in central Germany. Even in Israel, located about 5000 km away from Iceland, IN were as high as otherwise only during desert dust storms. These measurements are the only ones available on the properties of IN in the Eyjafjallajökull plume. The measured high concentrations and high activation temperature (−8 °C) point to an important impact of volcanic ash on microphysical and radiative properties of clouds through enhanced glaciation.