16 research outputs found
DaNa2.0 â Daten und Wissen zu Nanomaterialien: Aufbereitung gesellschaftlich relevanter naturwissenschaftlicher Fakten - Schlussbericht DaNa2.0 - Einzelantrag KIT
Das Projekt DaNa2.0 hatte zum Ziel, in einem interdisziplinĂ€ren Ansatz mit Wissenschaftlern aus Humantoxikologie, Ăkotoxikologie, Biologie, Physik, Chemie und Pharmazie Forschungsergebnisse zu Nanomaterialien und deren Auswirkungen auf den Menschen und die Umwelt fuÌr interessierte Verbraucher verstĂ€ndlich aufzubereiten. Diese Daten wurden auf der Internetplattform www.nanopartikel.info und auch durch andere Medien (Info-Flyer, BroschuÌren) der Ăffentlichkeit zugĂ€nglich gemacht.
Die wissenschaftlichen Ergebnisse der Projekte der BMBF-FördermaĂnahmen NanoCare, NanoNature, NanoCare4.0 und ERANET SIINN wurden als erweiterte Basis dieses Informationsangebots aufbereitet. Zudem wurde Literatur zu Human- und Umwelttoxikologie von Nanomaterialien auch anderer wissenschaftlicher Gruppen zur Erweiterung der Wissensplattform evaluiert. Die Bewertung der aktuellen Forschungsergebnisse fuÌr die DaNa-Wissensbasis erfolgt auf der Grundlage sorgfĂ€ltiger wissenschaftlicher Vorgehensweise anhand des DaNa Literatur-Kriterienkatalogs
The DaNa2.0 Knowledge Base Nanomaterials - An Important Measure Accompanying Nanomaterials Development
Nanotechnology is closely related to the tailored manufacturing of nanomaterials for a huge variety of applications. However, such applications with newly developed materials are also a reason for concern. The DaNa2.0 project provides information and support for these issues on the web in condensed and easy-to-understand wording. Thus, a key challenge in the field of advanced materials safety research is access to correct and reliable studies and validated results. For nanomaterials, there is currently a continuously increasing amount of publications on toxicological issues, but criteria to evaluate the quality of these studies are necessary to use them e.g., for regulatory purposes. DaNa2.0 discusses scientific results regarding 26 nanomaterials based on actual literature that has been selected after careful evaluation following a literature criteria checklist. This checklist is publicly available, along with a selection of standardized operating protocols (SOPs) established by different projects. The spectrum of information is rounded off by further articles concerning basics or crosscutting topics in nanosafety research. This article is intended to give an overview on DaNa2.0 activities to support reliable toxicity testing and science communication alik
Eight Weeks Later - The Unprecedented Rise of 3D Printing during the COVID-19 Pandemic : a Case Study, Lessons Learned, and Implications on the Future of Global Decentralized Manufacturing
The eruption of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (corona virus disease, COVID-19) in Wuhan, China, and its global spread has led to an exponentially growing number of infected patients, currently exceeding over 6.6 million and over 390,000 deaths as of the 5th of June 2020. In this pandemic situation, health systems have been put under stress, and the demand for personal protective equipment (PPE) exceeded the delivery capabilities of suppliers. To address this issue, 3D printing was identified as a possible solution to quickly produce PPE items such as face shields, mask straps, masks, valves, and ear savers. Around the world, companies, universities, research institutions, and private individuals/hobbyists stepped into the void, using their 3D printers to support hospitals, doctors, nursing homes, and even refugee camps by providing them with PPE. In Germany, the makervsvirus movement took up the challenge and connected thousands of end users, makers, companies, and logistic providers for the production and supply of face shields, protective masks, and ear savers. The Karlsruhe Institute of Technology (KIT) also joined the makervsvirus movement and used its facilities to print headbands for face shield assemblies and ear savers. Within this paper, the challenges and lessons learned from the quick ramp up of a research laboratory to a production site for medium-sized batches of PPE, the limitations in material supply, selection criteria for suitable models, quality measures, and future prospects are reported and conclusions drawn
Cerebrospinal fluid findings in adults with acute Lyme neuroborreliosis
Presence of BB-specific antibodies in the cerebrospinal fluid (CSF) with evidence of their intrathecal production in conjunction with the white cell count in the CSF and typical clinical symptoms is the traditional diagnostic gold standard of Lyme neuroborreliosis (LNB). Few data are available on the CSF lactate concentration in European adults with the diagnosis of acute LNB. The objective of the study was to investigate the CSF changes during acute LNB. Routine CSF parameters [leukocyte count, protein, lactate and albumin concentrations, CSF/serum quotients of albumin (QAlb), IgG, IgA and IgM, and oligoclonal IgG bands] and the Borrelia burgdorferi (BB)-specific antibody index were retrospectively studied in relation to the clinical presentation in patients diagnosed with acute LNB. A total of 118 patients with LNB were categorized into the following groups according to their symptoms at presentation; group 1: polyradiculoneuritis (Bannwarthâs syndrome), group 2: isolated facial palsy and group 3: predominantly meningitic course of the disease. In addition to the CSF of patients with acute LNB, CSF of 19 patients with viral meningitis (VM) and 3 with neurolues (NL) were analyzed. There were 97 patients classified with definite LNB, and 21 as probable LNB. Neck stiffness and fever were reported by 15.3% of patients. Most of these patients were younger than 50 years. Polyradiculoneuritis was frequently found in patients older than 50 years. Lymphopleocytosis was found in all patients. Only 5 patients had a CSF lactate â„3.5 mmol/l, and the mean CSF lactate level was not elevated (2.1 ± 0.6 mmol/l). The patients with definite LNB had significantly higher lactate levels than patients with probable LNB. Elevated lactate levels were accompanied by fever and headache. In the Reiber nomograms, intrathecal immunoglobulin synthesis was found for IgM in 70.2% followed by IgG in 19.5%. Isoelectric focussing detected an intrathecal IgG synthesis in 83 patients (70.3%). Elevated BB AIs in the CSF were found in 97 patients (82.2%). Patients with VM showed lower CSF protein concentration and CSF/serum quotients of albumin than LNB patients. In acute LNB, all patients had elevated cerebrospinal fluid (CSF) leukocyte counts. In contrast to infections by other bacteria, CSF lactate was lower than 3.5 mmol/l in all but 5 patients. The CSF findings did not differ between polyradiculoneuritis, facial palsy, and meningitis. The CSF in LNB patients strongly differed from CSF in VM patients with respect to protein concentration and the CSF/serum albumin quotient
CFC â Carbon Fibre Cycle Carbonfasern im Kreislauf â Freisetzungsverhalten und ToxizitĂ€t bei thermischer und mechanischer Behandlung - Poster-Abstracts
Identification and functional analysis of peroxisomal proteins
0 Titelblatt und Inhalt 1 Einleitung 1
2 Material und Methoden 15
3 Ergebnisse 38
4 Diskussion 113
5 Zusammenfassung 137
6 Abstract 139
7 Literatur 141
8 Anhang 152Identifizierung In dieser Arbeit wurden unter Verwendung des ursprĂŒnglichen
reversen genetischen Ansatzes zur Identifizierung peroxisomaler Proteine 78
Proteinbanden massenspektrometrisch analysiert und dabei 59 verschiedene
Proteine identifiziert. Neunzehn der Proteine erwiesen sich als peroxisomal,
darunter auch die neu identifizierten Proteine Eci1p (Enoyl-CoA Isomerase),
Dci1p (Dienoyl-CoA Isomerase), welchen eine akzessorische Funktion beim Abbau
ungesĂ€ttigter FettsĂ€uren zukommt, und die peroxisomale Thioesterase Tes1p. FĂŒr
elf weitere Proteine dieses Ansatzes konnte eine potentielle peroxisomale
Lokalisation angenommen werden. Im Rahmen dieser Studie wurde ein zweiter,
modifizierter Ansatz zur Analyse des peroxisomalen Proteoms etabliert, der auf
dem kombinatorischen Einsatz von 1D-SDS-Gelelektrophorese und MALDI-MS und LC-
ESI-MS basierte. Hierbei konnten 66 Proteine identifiziert werden, darunter
befanden sich 25 peroxisomale. 17 Proteine, deren Lokalisation in der Zelle
noch nicht festgestellt worden war, wurden daraufhin untersucht.
Funktionsanalyse Die subzellulÀre Lokalisation unbekannter Proteine wurde mit
Hilfe von Doppel-Fluoreszenzmikroskopie unter Verwendung von GFP-
Fusionsproteinen in Verbindung mit einem rot-fluoreszierenden peroxisomalen
Markerprotein (PTS2DsRed) und Zellfraktionierungsanalysen untersucht. Zur
KlÀrung der potentiellen peroxisomalen Funktion eines neuen Proteins, wurde
das Wachstum der entsprechenden Deletionsmutante auf verschiedenen
Kohlenstoffquellen analysiert. Es konnte gezeigt werden, dass Yor084wp eine
ölsÀureinduzierbare, potentielle peroxisomale Lipase/Esterase ist, die Pex5p-
abhÀngig in die Peroxisomenmatrix importiert wird. Die Deletion des
entsprechenden Gens hat keinen Einfluss auf die Degradation von FettsÀuren.
Die potentielle 2-Hydroxyphytanoyl-CoA-Lyase Yel020cp kann zu den
peroxisomalen Matrixproteinen gezÀhlt werden, die Pex5p-abhÀngig importiert
werden. Sie verfĂŒgt auĂerdem ĂŒber eine Thiamin-Diphosphat-Bindestelle. FĂŒr die
Biogenese von Peroxisomen ist das Protein nicht essentiell. Ygl184cp ist eine
mögliche Cystathionin-b-Lyase und konnte in dieser Arbeit in der Membran von
Peroxisomen nachgewiesen werden. FĂŒr die Biogenese der Organellen wird dieses
Protein nicht benötigt. Yir034cp ist ein peroxisomales Matrixprotein, das
aufgrund seiner PTS1-Sequenz in die Organellen gelangt. Seine Rolle in der
Biosynthese des Lysins fĂŒhrte zu der Hypothese, dass dieser
AminosĂ€urestoffwechsel in Hefe in Peroxisomen stattfindet. FĂŒr die Acetyl-CoA-
Hydrolase Ybl015wp konnte gezeigt werden, dass sie aufgrund ihrer potentiellen
N-terminalen peroxisomalen (PTS2) als auch mitochondrialen
Zielsteuerungssequenz (MTS) in beide Organellen importiert wird. Ycr091wp
wurde ebenfalls in diesen beiden Organellen nachgewiesen und sorgt dort als
potentielle Serin/Threonin-Kinase vermutlich fĂŒr entsprechende
posttranslationale Modifikationen. Die mitochondriale Lokalisation des
unbekannten Proteins Yor228cp konnte gezeigt werden. ComputergestĂŒtzte
Analysen weisen auĂerdem auf drei transmembrane Bereiche hin, so dass es sich
um ein mitochondriales Membranprotein handeln könnte. Der Succinat-Fumarat-
Transporter Sfc1p/Yjr095wp gehört zur Familie der mitochondrialen Transporter,
der auch peroxisomale Proteine angehören. Eine zusÀtzliche peroxisomale
Lokalisation konnte fĂŒr dieses Protein nicht bestimmt werden. Das unbekannte
Yhr199cp wurde ebenfalls in den Mitochondrien detektiert, die zellulÀre
Funktion bleibt allerdings unbekannt. Die vermutete peroxisomale Lokalisation
des Yal054cp konnte nicht bestÀtigt werden. Dagegen wurde eine mikrosomale
Lokalisation vermutet. FĂŒr weitere 22 untersuchte Proteine konnte ebenfalls
keine peroxisomale Lokalisation festgestellt werden, so dass diese Proteine
als Kontaminationen bei der Isolierung peroxisomaler Proteine betrachtet
werden mĂŒssen. Pex11 Im Rahmen dieser Arbeit konnte mittels
elektrophysiologischer Messungen nicht nachgewiesen werden, ob Pex11p einen
spannungsgesteuerten Kanal in der Peroxisomenmembran bildet. ScPex11p wurde in
Form eines TAP-Fusionsproteins erfolgreich exprimiert und kann so zur
affinitĂ€tschromatographischen Isolierung und weiterfĂŒhrenden Funktionsanalysen
verwendet werden.Identification In this study, a reverse genetic screen for peroxisomal
proteins was used to analyze 78 protein bands of SDS-PAGE by mass
spectrometry. 59 different proteins could be identified. 19 of the proteins
proved to be peroxisomal, among them the 3 newly identified proteins Eci1p
(enoyl-CoA isomerase), Dci1p (dienoyl-CoA isomerase) which play an accessory
role in degradation of unsaturated fatty acids and Tes1p (thioesterase). 11
unknown proteins were suspected to be peroxisomal. In this study, an
alternative approach for the analysis of the peroxisomal proteome was
established, which combines protein separation during a 1-D SDS-PAGE with
MALDI-MS and LC-ESI-MS. It was possible to identify 66 proteins, 25
peroxisomal and 17 proteins of not yet known localization Functional analysis
The localization of newly identified proteins was confirmed by double-
fluorescence microscopy using GFP fusion proteins in conjunction with a
peroxisomal red fluorescent protein marker (PTS2DsRed) as well as by
subcellular fractionation studies. To reveal the function of newly identified
proteins, corresponding knockout strains were analyzed phenotypically for
growth on various carbon sources. It was shown, that Yor084wp functions as an
oleic acid-inducible putative peroxisomal lipase/esterase. Import in the
peroxisomal membrane is Pex5p-dependent. The observed phenotype of the
analyzed mutant allows the conclusions that the corresponding gene has no
effect of fatty acid degradation. The putative 2-hydroxyphytanoyl CoA lyase
Yel020cp belongs to the peroxisomal, Pex5p-dependent imported matrix proteins.
The protein possesses a thiamin-pyrophosphate-binding site. The gene seems to
be not essential for peroxisomal biogenesis. Ygl184cp functions as a putative
cystathionine b-lyase and in this study it was detected at the peroxisomal
membrane. Yir034cp is a protein of the peroxisomal matrix that is imported in
the organelle because of its PTS1-sequence. Its role in the biosynthesis of
lysine leads to the hypothesis, that this kind of amino acid pathway is
localized in yeast peroxisomes. For the acetyl-CoA hydrolase Ybl015wp, it was
shown that it is imported in peroxisomes and in mitochondria because of the
potential N-terminal peroxisomal (PTS2) as well as the mitochondrial targeting
signal (MTS). Ycr091wp is also present in mitochondria and peroxisomes and
functions presumably as a putative serine/threonine kinase in
posttranslational modifications. In this study, the mitochondrial localization
of the unknown protein Yor228cp was shown and three transmembrane domains were
detected by in silico analysis, that led to the assumption that it is
localized in the mitochondrial membrane. Sfc1p/Yjr095wp is a member of the
mitochondrial carrier family (MCF) and works as a succinate-fumarate
transporter. A second, peroxisomal localization was not found. Yhr199cp was
also found in mitochondria because of a mitochondria targeting signal, but its
cellular role was not analyzed. The suspected peroxisomal localization of the
Yal054cp could be disproved. The observed subcellular localization gave rise
to the suspicion that it is localized in ER. 22 proteins were not peroxisomal,
indicating that minor contaminations of the preparation were also detected due
to the sensitivity of the method. Pex11p In this study, the putative function
of Pex11p as a voltage-dependent channel in the peroxisomal membrane could not
be demonstrated. The expression of the yeast Pex11p as a TAP-fusion protein
was successful and the protein can be purified by affinity columns and than
used for further functional analysis
The DaNa2.0 Knowledge Base NanomaterialsâAn Important Measure Accompanying Nanomaterials Development
Nanotechnology is closely related to the tailored manufacturing of nanomaterials for a huge variety of applications. However, such applications with newly developed materials are also a reason for concern. The DaNa2.0 project provides information and support for these issues on the web in condensed and easy-to-understand wording. Thus, a key challenge in the field of advanced materials safety research is access to correct and reliable studies and validated results. For nanomaterials, there is currently a continuously increasing amount of publications on toxicological issues, but criteria to evaluate the quality of these studies are necessary to use them e.g., for regulatory purposes. DaNa2.0 discusses scientific results regarding 26 nanomaterials based on actual literature that has been selected after careful evaluation following a literature criteria checklist. This checklist is publicly available, along with a selection of standardized operating protocols (SOPs) established by different projects. The spectrum of information is rounded off by further articles concerning basics or crosscutting topics in nanosafety research. This article is intended to give an overview on DaNa2.0 activities to support reliable toxicity testing and science communication alike