TY - JOUR A1 - Reyes, Anibal M. A1 - Vazquez, Diego S. A1 - Zeida, Ari A1 - Hugo, Martin A1 - Dolores Pineyro, M. A1 - Ines De Armas, Maria A1 - Estrin, Dario A1 - Radi, Rafael A1 - Santos, Javier A1 - Trujillo, Madia T1 - PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase JF - Free radical biology and medicine : the official journal of the Oxygen Society, a constituent member of the International Society for Free Radical Research N2 - Mycobacterium tuberculosis (M. tuberculosis) is the intracellular bacterium responsible for tuberculosis disease (TD). Inside the phagosomes of activated macrophages, M. tuberculosis is exposed to cytotoxic hydroperoxides such as hydrogen peroxide, fatty acid hydroperoxides and peroxynitrite. Thus, the characterization of the bacterial antioxidant systems could facilitate novel drug developments. In this work, we characterized the product of the gene Rv1608c from M. tuberculosis, which according to sequence homology had been annotated as a putative peroxiredoxin of the peroxiredoxin Q subfamily (PrxQ B from M. tuberculosis or MtPrxQ B). The protein has been reported to be essential for M. tuberculosis growth in cholesterol-rich medium. We demonstrated the M. tuberculosis thioredoxin B/C-dependent peroxidase activity of MtPrxQ B, which acted as a two-cysteine peroxiredoxin that could function, although less efficiently, using a one-cysteine mechanism. Through steady-state and competition kinetic analysis, we proved that the net forward rate constant of MtPrxQ B reaction was 3 orders of magnitude faster for fatty acid hydroperoxides than for hydrogen peroxide (3x10(6) vs 6x10(3) M-1 s(-1), respectively), while the rate constant of peroxynitrite reduction was (0.6-1.4) x10(6) M-1 s(-1) at pH 7.4. The enzyme lacked activity towards cholesterol hydroperoxides solubilized in sodium deoxycholate. Both thioredoxin B and C rapidly reduced the oxidized form of MtPrxQ B, with rates constants of 0.5x10(6) and 1x10(6) M-1 s(-1), respectively. Our data indicated that MtPrxQ B is monomeric in solution both under reduced and oxidized states. In spite of the similar hydrodynamic behavior the reduced and oxidized forms of the protein showed important structural differences that were reflected in the protein circular dichroism spectra. KW - Mycobacterium tuberculosis KW - Peroxiredoxin KW - Thioredoxin KW - Peroxynitrite KW - Fatty acid hydroperoxides KW - Thiol-dependent peroxidase KW - Peroxidatic and resolving cysteine Y1 - 2016 U6 - https://doi.org/10.1016/j.freeradbiomed.2016.10.005 SN - 0891-5849 SN - 1873-4596 VL - 101 SP - 249 EP - 260 PB - Elsevier CY - New York ER - TY - THES A1 - Stolzenburg, Antje T1 - Bittergeschmacksrezeptoren des peripheren und zentralen Nervensystems T1 - Bitter taste receptors of the peripheral and central nervous system N2 - Der Bittergeschmack warnt den Organismus vor potentiell verdorbener oder giftiger Nahrung und ist somit ein wichtiger Kontrollmechanismus. Die initiale Detektion der zahlreich vorkommenden Bitterstoffe erfolgt bei der Maus durch 35 Bitterrezeptoren (Tas2rs), die sich im Zungengewebe befinden. Die Geschmacksinformation wird anschließend von der Zunge über das periphere (PNS) ins zentrale Nervensystem (ZNS) geleitet, wo deren Verarbeitung stattfindet. Die Verarbeitung der Geschmacksinformation konnte bislang nicht gänzlich aufgeklärt werden. Neue Studien deuten auf eine Expression von Tas2rs auch im PNS und ZNS entlang der Geschmacksbahn hin. Über Vorkommen und Aufgaben dieser Rezeptoren bzw. Rezeptorzellen im Nervensystem ist bislang wenig bekannt. Im Rahmen dieser Arbeit wurde die Tas2r-Expression in verschiedenen Mausmodellen untersucht, Tas2r-exprimierende Zellen identifiziert und deren Funktionen bei der Übertragung der Geschmacksinformationen analysiert. Im Zuge der Expressionsanalysen mittels qRT-PCR konnte die Expression von 25 der 35 bekannten Bittergeschmacksrezeptoren im zentralen Nervensystem der Maus nachgewiesen werden. Die Expressionsmuster im PNS sowie im ZNS lassen darüber hinaus Vermutungen zu Funktionen in verschiedenen Bereichen des Nervensystems zu. Basierend auf den Ergebnissen der Expressionsanalysen war es möglich, stark exprimierte Tas2rs mittels In-situ-Hybridisierung in verschiedenen Zelltypen zu visualisieren. Des Weiteren konnten immunhistochemische Färbungen unter Verwendung eines genetisch modifizierten Mausmodells die Ergebnisse der Expressionsanalysen bestätigen. Sie zeigten eine Expression von Tas2rs, am Beispiel des Tas2r131-Rezeptors, in cholinergen, dopaminergen, GABAergen, noradrenergen und glycinerg-angesteuerten Projektionsneuronen sowie in Interneuronen. Die Ergebnisse der vorliegenden Arbeit zeigen daher erstmals das Vorkommen von Tas2rs in verschiedenen neuronalen Zelltypen in weiten Teilen des ZNS. Dies lässt den Schluss zu, dass Tas2r-exprimierende Zellen potentiell multiple Funktionen innehaben. Anhand von Verhaltensexperimenten in genetisch modifizierten Mäusen wurde die mögliche Funktion von Tas2r131-exprimierenden Neuronen (Tas2r131-Neurone) bei der Geschmackswahrnehmung untersucht. Die Ergebnisse weisen auf eine Beteiligung von Tas2r131-Neuronen an der Signalweiterleitung bzw. -verarbeitung der Geschmacksinformation für eine Auswahl von Bittersubstanzen hin. Die Analysen zeigen darüber hinaus, dass Tas2r131-Neuronen nicht an der Geschmackswahrnehmung anderer Bitterstoffe sowie Geschmacksstimuli anderer Qualitäten (süß, umami, sauer, salzig), beteiligt sind. Eine spezifische „Tas2r131-Bittergeschmacksbahn“, die mit anderen potentiellen „Bitterbahnen“ teils unabhängige, teils überlappende Signalwege bzw. Verarbeitungsbereiche besitzt, bildet eine mögliche zelluläre Grundlage zur Unterscheidung von Bitterstoffen. Die im Rahmen dieser Arbeit entstandene Hypothese einer potentiellen Diskriminierung von Bitterstoffen soll daher in weiterführenden Studien durch die Etablierung eines Verhaltenstest mit Mäusen geprüft werden. N2 - Bitter taste warns the organism about potentially spoiled or toxic food and is thus an important control mechanism. The initial detection of numerous occurring bitter substances is done in mice by 35 bitter taste receptors (Tas2rs), located in tongue tissue. From the tongue the gustatory information is then passed via the peripheral (PNS) to the central nervous system (CNS), where it is processed. The processing of taste information couldn’t yet be clarified entirely. Recent studies point to an expression of Tas2rs also in the PNS and CNS along the taste transmission pathway. However, little is known concerning occurrence and functions of Tas2rs or Tas2r-expressing cells in the nervous system. In this work the Tas2r expression was examined in different mouse models, Tas2r-expressing cells were identified and their functions in transmission of taste information analyzed. Expression analyses using qRT-PCR showed an expression of 25 of the 35 known murine bitter taste receptors in the central nervous system. The expression patterns in the PNS and CNS suggests functions in different areas of the nervous system. Based on the results of the expression analysis it was possible to visualize highly expressed Tas2rs by in-situ-hybridization in various cell types. Furthermore, immunohistochemical staining using a genetically modified mouse model confirmed the results of the expression analysis. They showed an expression of Tas2rs, on the example of the Tas2r131 receptor, in the cholinergic, dopaminergic, GABAergic, noradrenergic and glycinerg-driven projection neurons and interneurons. The results of the present work show for the first time the presence of Tas2rs in different neuronal cell types in many parts of the CNS. This leads to the conclusion that Tas2r-expressing cells hold potentially multiple functions. Based on behavioral experiments in genetically modified mice, the possible taste function of Tas2r131-expressing neurons (Tas2r131 neurons) was studied. The results showed the involvement of Tas2r131 neurons in signal transduction and processing of gustatory information for a selection of bitter substances. Besides, the analyses show that Tas2r131 neurons aren’t involved in taste perception for another selection of bitter substances and taste stimuli of other qualities (sweet, umami, sour, salty). A specific "Tas2r131-bitter-pathway" which forms partly independent and partly overlapping signaling pathways or processing areas with other potential "bitter-pathways", provides a cellular basis for the distinction of specific bitter compounds. The resulting hypothesis of a potential discrimination of bitter substances should therefore be examined in further studies by establishing a behavioral test with mice. KW - Geschmack KW - Bittergeschmack KW - Bittergeschmacksrezeptoren KW - Tas2r KW - Verhaltensstudien KW - zentrales Nervensystem KW - peripheres Nervensystem KW - Neurone KW - Maus KW - taste KW - bitter taste KW - bitter taste receptors KW - Tas2rs KW - peripheral nervous system KW - central nervous system KW - neuron KW - behavioral experiments KW - expression analysis KW - mouse Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-92397 ER - TY - THES A1 - Kamitz, Anne T1 - Identification and positional cloning of Ltg/NZO; a novel susceptibility locus associated with fatty liver disease Y1 - 2016 ER - TY - THES A1 - Töle, Nadine T1 - Molekulare und histologische Untersuchungen zur gustatorischen Fettwahrnehmung des Menschen T1 - Molecular and histological analyses of human gustatory fat perception N2 - Die hohe Energieaufnahme durch Fette ist ein Hauptfaktor für die Entstehung von Adipositas, was zu weltweiten Bestrebungen führte, die Fettaufnahme zu verringern. Fettreduzierte Lebensmittel erreichen jedoch, trotz ihrer Weiterentwicklung, nicht die Schmackhaftigkeit ihrer Originale. Die traditionelle Sichtweise, dass die Attraktivität von Fetten allein durch Textur, Geruch, Aussehen und postingestive Effekte bestimmt wird, wird nun durch das Konzept einer gustatorischen Wahrnehmung ergänzt. Bei Nagetieren zeigte sich, dass Lipide unabhängig von den vorgenannten Eigenschaften erkannt werden, sowie, dass Fettsäuren, freigesetzt durch linguale Lipasen, als gustatorische Stimuli fungieren und Fettsäuresensoren in Geschmackszellen exprimiert sind. Die Datenlage für den Menschen erwies sich jedoch als sehr begrenzt, daher war es Ziel der vorliegenden Arbeit molekulare und histologische Voraussetzungen für eine gustatorische Fettwahrnehmung beim Menschen zu untersuchen. Zunächst wurde humanes Geschmacksgewebe mittels RT-PCR und immunhistochemischen Methoden auf die Expression von Fettsäuresensoren untersucht, sowie exprimierende Zellen in Kofärbeexperimenten charakterisiert und quantifiziert. Es wurde die Expression fettsäuresensitiver Rezeptoren nachgewiesen, deren Agonisten das gesamte Spektrum an kurz- bis langkettigen Fettsäuren abdecken (GPR43, GPR84, GPR120, CD36, KCNA5). Ein zweifelsfreier Nachweis des Proteins konnte für den auf langkettige Fettsäuren spezialisierten Rezeptor GPR120 in Typ-I- und Typ-III-Geschmackszellen der Wallpapillen erbracht werden. Etwa 85 % dieser GPR120-exprimierenden Zellen enthielten keine der ausgewählten Rezeptoren der Geschmacksqualitäten süß (TAS1R2/3), umami (TAS1R1/3) oder bitter (TAS2R38). Somit findet sich in humanen Geschmackspapillen nicht nur mindestens ein Sensor, sondern möglicherweise auch eine spezifische, fettsäuresensitive Zellpopulation. Weitere RT-PCR-Experimente und Untersuchungen mittels In-situ-Hybridisierung wurden zur Klärung der Frage durchgeführt, ob Lipasen in den Von-Ebner-Speicheldrüsen (VED) existieren, die freie Fettsäuren aus Triglyceriden als gustatorischen Stimulus freisetzen können. Es zeigte sich zwar keine Expression der bei Nagetieren gefundenen Lipase F (LIPF), jedoch der eng verwandten Lipasen K, M und N in den serösen Zellen der VED. In-silico-Untersuchungen der Sekundär- und Tertiärstrukturen zeigten die hohe Ähnlichkeit zu LIPF, erwiesen aber auch Unterschiede in den Bindungstaschen der Enzyme, welche auf ein differenziertes Substratspektrum hinweisen. Die Anwesenheit eines spezifischen Signalpeptids macht eine Sekretion der Lipasen in den die Geschmacksporen umspülenden Speichel wahrscheinlich und damit auch eine Bereitstellung von Fettsäuren als Stimuli für Fettsäuresensoren. Die Übertragung des durch diese Stimuli hervorgerufenen Signals von Geschmackszellen auf gustatorische Nervenfasern über P2X-Rezeptormultimere wurde mit Hilfe einer vorherigen Intervention mit einem P2X3 /P2X2/3-spezifischen Antagonisten an der Maus als Modellorganismus im Kurzzeit-Präferenztest untersucht. Es zeigte sich weder eine Beeinträchtigung der Wahrnehmung einer Fettsäurelösung, noch einer zuckerhaltigen Kontrolllösung, wohingegen die Wahrnehmung einer Bitterstofflösung reduziert wurde. Somit ist anhand der Ergebnisse dieser Arbeit eine Beteiligung des P2X3-Homomers bzw. des P2X2/3-Heteromers unwahrscheinlich, jedoch die des P2X2-Homomers und damit der gustatorischen Nervenfasern nicht ausgeschlossen. Die Ergebnisse dieser Arbeit weisen auf die Erfüllung grundlegender Voraussetzungen für die gustatorische Fett(säure)wahrnehmung hin und tragen zum Verständnis der sensorischen Fettwahrnehmung und der Regulation der Fettaufnahme bei. Das Wissen um die Regulation dieser Mechanismen stellt eine Grundlage zur Aufklärung der Ursachen und damit der Bekämpfung von Adipositas und assoziierten Krankheiten dar. N2 - High consumption of energy from fat is considered one of the main factors that evoke obesity which led to a worldwide effort to reduce dietary fat intake. However, despite their continuous improvement, fat-reduced foods do not yet reach the palatability of their originals. The traditional view that the attraction to fats is only determined by texture, odor, appearance as well as postingestive effects is now challenged by the concept of gustatory sensation of fats. After excluding or masking the aforementioned features, rodents showed continuous attraction towards lipid solutions. Also long-chain fatty acids liberated from triglycerides by lingual lipases as the main stimulus and the expression of fatty acid-sensitive receptors in taste buds was shown. In contrast, only little data exists for humans. Therefore, this thesis aimed at elucidating the molecular and cellular prerequisites for a gustatory detection of fat. First, human taste tissue was examined by RT-PCR and immunohistochemical methods for the expression of fatty acid sensors and the expressing cells were characterized and quantified by co-staining procedures. The expression of fatty acid-sensitive receptors was shown whose agonists cover the whole range of short- to long-chain fatty acids (GPR43, GPR84, GPR120, CD36, and KCNA5). Protein expression of the long-chain fatty acid receptor GPR120 in type I and type III taste cells was unambiguously demonstrated. About 85 % of the GPR120-expressing cells did not co-express receptors for sweet (TAS1R2/3), umami (TAS1R1/3) or bitter (TAS2R38) taste. Hence, not only does at least one fatty acid sensor exist in human taste papillae, but possibly also a special fatty acid-sensitive cell population. Additional RT-PCR experiments and in situ hybridization analyses were used to address the question, if lipases in the Von-Ebner-salivary glands (VEG) produce free fatty acids from triglycerides as gustatory stimuli. Unlike mice which express lipase F, lipases K, M and N, which are highly related to lipase F, were found to be expressed in the serous cells of the VEG. In silico approaches confirmed high similarities of the secondary and tertiary structures of these lipases to lipase F. Also a marked difference in the binding pocket of the enzymes was observed, which suggests differential substrate specificity. The presence of a signal peptide sequence proposes that the lipases K, M, and N are secreted into the trenches of gustatory papillae. This would result in lipolysis of dietary triglycerides and generation of stimuli for the fatty acid sensor GPR120. Next the hypothesis was tested whether GPR120-generated signals are conveyed to gustatory nerves via P2X-receptor-multimers. To this end short term preference tests were performed in mice after intervention with a P2X3-/P2X2/3-specific antagonist. However, this treatment did not affect the preference of mice for the fatty acid or for a sweet control solution, whereas recognition of a bitter solution was impaired. Thus, an involvement of the P2X3-homomer or the P2X3-/P2X2/3-heteromer seems unlikely. However, these results do not exclude a contribution of P2X2-receptor-heteromers and in turn gustatory nerves for the preference of fatty acid solutions. In summary, the results of this thesis give new insights into the molecular and cellular prerequisites for a gustatory component for fat detection in humans. They also help understanding fat sensation and the regulation of fat intake which in turn may eventually promote novel concepts for the treatment of obesity and related diseases. KW - Geschmack KW - Fettwahrnehmung KW - Fettsäuren KW - Sensorik KW - Lipasen KW - G-Protein gekoppelte Rezeptoren KW - taste KW - fat perception KW - fatty acids KW - lipases KW - G-protein coupled receptors KW - sensory analysis Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-93180 ER - TY - JOUR A1 - Reinkensmeier, Annika A1 - Steinbrenner, Katrin A1 - Homann, Thomas A1 - Bussler, Sara A1 - Rohn, Sascha A1 - Rawel, Harshadrai Manilal T1 - Monitoring the apple polyphenol oxidase-modulated adduct formation of phenolic and amino compounds JF - Food chemistry N2 - Minimally processed fruit products such as smoothies are increasingly coming into demand. However, they are often combined with dairy ingredients. In this combination, phenolic compounds, polyphenoloxidases, and amino compounds could interact. In this work, a model approach is presented where apple serves as a source for a high polyphenoloxidase activity for modulating the reactions. The polyphenoloxidase activity ranged from 128 to 333 nakt/mL in different apple varieties. From these, ‘Braeburn’ was found to provide the highest enzymatic activity. The formation and stability of resulting chromogenic conjugates was investigated. The results show that such adducts are not stable and possible degradation mechanisms leading to follow-up products formed are proposed. Finally, apple extracts were used to modify proteins and their functional properties characterized. There were retaining antioxidant properties inherent to phenolic compounds after adduct formation. Consequently, such interactions may also be utilized to improve the textural quality of food products. KW - Apple polyphenoloxidase KW - Phenol-amino-adducts KW - Post-translational protein modification KW - Functionality Y1 - 2016 U6 - https://doi.org/10.1016/j.foodchem.2015.07.145 SN - 0308-8146 SN - 1873-7072 VL - 194 SP - 76 EP - 85 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Bußler, Sara A1 - Rumpold, Birgit A. A1 - Fröhling, Antje A1 - Jander, Elisabeth A1 - Rawel, Harshadrai Manilal A1 - Schlüter, Oliver K. T1 - Cold atmospheric pressure plasma processing of insect flour from Tenebrio molitor: Impact on microbial load and quality attributes in comparison to dry heat treatment JF - Meteoritics & planetary science : journal of the Meteoritical Society N2 - In this study, the applicability of semi-direct cold atmospheric pressure plasma (CAPP) during postharvest processing of Tenebrio molitor flour is investigated. Besides analyzing the decontamination efficacy, plasma induced impact on techno-functionality, protein solubility, composition and structure was determined and compared to heat induced effects. Following CAPP treatment, the total microbial load of the Tenebrio flour of 7.72 log(10) cfu/g was reduced to 7.10 (1 min), 6.72 (2.5 min), 5.79 (5 min), 5.19 (7.5 min), 521 (10 min) and 4.73 (15 min) log(10) cfu/g. With increasing exposure to CAPP, protein solubility at pH 4 almost linearly decreased to a minimum of 54%. Water binding capacity decreased from 0.79 to 0.64 gwatedg whereas oil binding capacity increased from 0.59 to 0.66 g(oil)/g. Gel electrophoresis revealed a decrease of all protein fractions at pH 4 whereas at pH 10 the band pattern significantly shifted to protein fractions with higher molecular weights. Industrial relevance: Edible insects are rich in valuable protein, fat, fibre, minerals and micronutrients. Although a wide range of species represent a valuable alternative protein source that could contribute to food and feed security, they are industrially hardly exploited. The tailored application of proper processing technologies could lead to novel insect-based high-protein food and feed products with unique functional properties supporting the increase in acceptability among potential consumers. Current research concentrates on developing processing chains including innovative nonthermal approaches. Cold atmospheric pressure plasma (CAPP) has gained attention as an effective technology for the decontamination and modification of fresh and dry agricultural products. In the postharvest chain of edible insects, the application of CAPP could contribute to the development of safe and high-quality insect-based products in the food and feed sector. (C) 2016 Published by Elsevier Ltd. KW - Edible insects KW - Postharvest processing KW - Thermal and nonthermal treatment KW - Inactivation KW - Decontamination KW - Protein functionality and modification Y1 - 2016 U6 - https://doi.org/10.1016/j.ifset.2016.07.002 SN - 1466-8564 SN - 1878-5522 VL - 36 SP - 277 EP - 286 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Reichetzeder, Christoph A1 - Putra, S. E. Dwi A1 - Pfab, T. A1 - Slowinski, T. A1 - Neuber, Corinna A1 - Kleuser, Burkhard A1 - Hocher, Berthold T1 - Increased global placental DNA methylation levels are associated with gestational diabetes JF - Clinical epigenetics N2 - Background: Gestational diabetes mellitus (GDM) is associated with adverse pregnancy outcomes. It is known that GDM is associated with an altered placental function and changes in placental gene regulation. More recent studies demonstrated an involvement of epigenetic mechanisms. So far, the focus regarding placental epigenetic changes in GDM was set on gene-specific DNA methylation analyses. Studies that robustly investigated placental global DNA methylation are lacking. However, several studies showed that tissue-specific alterations in global DNA methylation are independently associated with type 2 diabetes. Thus, the aim of this study was to characterize global placental DNA methylation by robustly measuring placental DNA 5-methylcytosine (5mC) content and to examine whether differences in placental global DNA methylation are associated with GDM. Methods: Global DNA methylation was quantified by the current gold standard method, LC-MS/MS. In total, 1030 placental samples were analyzed in this single-center birth cohort study. Results: Mothers with GDM displayed a significantly increased global placental DNA methylation (3.22 +/- 0.63 vs. 3.00 +/- 0.46 %; p = 0.013; +/- SD). Bivariate logistic regression showed a highly significant positive correlation between global placental DNA methylation and the presence of GDM (p = 0.0009). Quintile stratification according to placental DNA 5mC levels revealed that the frequency of GDM was evenly distributed in quintiles 1-4 (2.9-5.3 %), whereas the frequency in the fifth quintile was significantly higher (10.7 %; p = 0.003). Bivariate logistic models adjusted for maternal age, BMI, ethnicity, recurrent miscarriages, and familiar diabetes predisposition clearly demonstrated an independent association between global placental DNA hypermethylation and GDM. Furthermore, an ANCOVA model considering known predictors of DNA methylation substantiated an independent association between GDM and placental DNA methylation. Conclusions: This is the first study that employed a robust quantitative assessment of placental global DNA methylation in over a thousand placental samples. The study provides large scale evidence that placental global DNA hypermethylation is associated with GDM, independent of established risk factors. KW - Placenta KW - Gestational diabetes KW - Insulin resistance KW - LC-MS/MS KW - Global DNA methylation KW - Epigenetics KW - Hypermethylation Y1 - 2016 U6 - https://doi.org/10.1186/s13148-016-0247-9 SN - 1868-7083 VL - 8 PB - BioMed Central CY - London ER - TY - JOUR A1 - Hocher, Berthold A1 - Haumann, Hannah A1 - Rahnenführer, Jan A1 - Reichetzeder, Christoph A1 - Kalk, Philipp A1 - Pfab, Thiemo A1 - Tsuprykov, Oleg A1 - Winter, Stefan A1 - Hofmann, Ute A1 - Li, Jian A1 - Püschel, Gerhard Paul A1 - Lang, Florian A1 - Schuppan, Detlef A1 - Schwab, Matthias A1 - Schaeffeler, Elke T1 - Maternal eNOS deficiency determines a fatty liver phenotype of the offspring in a sex dependent manner JF - Epigenetics : the official journal of the DNA Methylation Society N2 - Maternal environmental factors can impact on the phenotype of the offspring via the induction of epigenetic adaptive mechanisms. The advanced fetal programming hypothesis proposes that maternal genetic variants may influence the offspring's phenotype indirectly via epigenetic modification, despite the absence of a primary genetic defect. To test this hypothesis, heterozygous female eNOS knockout mice and wild type mice were bred with male wild type mice. We then assessed the impact of maternal eNOS deficiency on the liver phenotype of wild type offspring. Birth weight of male wild type offspring born to female heterozygous eNOS knockout mice was reduced compared to offspring of wild type mice. Moreover, the offspring displayed a sex specific liver phenotype, with an increased liver weight, due to steatosis. This was accompanied by sex specific differences in expression and DNA methylation of distinct genes. Liver global DNA methylation was significantly enhanced in both male and female offspring. Also, hepatic parameters of carbohydrate metabolism were reduced in male and female offspring. In addition, male mice displayed reductions in various amino acids in the liver. Maternal genetic alterations, such as partial deletion of the eNOS gene, can affect liver metabolism of wild type offspring without transmission of the intrinsic defect. This occurs in a sex specific way, with more detrimental effects in females. This finding demonstrates that a maternal genetic defect can epigenetically alter the phenotype of the offspring, without inheritance of the defect itself. Importantly, these acquired epigenetic phenotypic changes can persist into adulthood. KW - Epigenetics KW - eNOS KW - Fetal programming KW - fatty liver KW - metabolism Y1 - 2016 U6 - https://doi.org/10.1080/15592294.2016.1184800 SN - 1559-2294 SN - 1559-2308 VL - 11 SP - 539 EP - 552 PB - Routledge, Taylor & Francis Group CY - Philadelphia ER - TY - JOUR A1 - Manowsky, Julia A1 - Camargo, Rodolfo Gonzalez A1 - Kipp, Anna Patricia A1 - Henkel, Janin A1 - Püschel, Gerhard Paul T1 - Insulin-induced cytokine production in macrophages causes insulin resistance in hepatocytes JF - American journal of physiology : Endocrinology and metabolism N2 - Overweight and obesity are associated with hyperinsulinemia, insulin resistance, and a low-grade inflammation. Although hyperinsulinemia is generally thought to result from an attempt of the beta-cell to compensate for insulin resistance, there is evidence that hyperinsulinaemia itself may contribute to the development of insulin resistance and possibly the low-grade inflammation. To test this hypothesis, U937 macrophages were exposed to insulin. In these cells, insulin induced expression of the proinflammatory cytokines IL-1 beta, IL-8, CCL2, and OSM. The insulin-elicited induction of IL-1 beta was independent of the presence of endotoxin and most likely mediated by an insulin-dependent activation of NF-kappa B. Supernatants of the insulin-treated U937 macrophages rendered primary cultures of rat hepatocytes insulin resistant; they attenuated the insulin-dependent induction of glucokinase by 50%. The cytokines contained in the supernatants of insulin-treated U937 macrophages activated ERK1/2 and IKK beta, resulting in an inhibitory serine phosphorylation of the insulin receptor substrate. In addition, STAT3 was activated and SOCS3 induced, further contributing to the interruption of the insulin receptor signal chain in hepatocytes. These results indicate that hyperinsulinemia per se might contribute to the low-grade inflammation prevailing in overweight and obese patients and thereby promote the development of insulin resistance particularly in the liver, because the insulin concentration in the portal circulation is much higher than in all other tissues. KW - metabolic syndrome KW - type 2 diabetes KW - inflammation KW - macrophage KW - insulin KW - cytokines Y1 - 2016 U6 - https://doi.org/10.1152/ajpendo.00427.2015 SN - 0193-1849 SN - 1522-1555 VL - 310 SP - E938 EP - E946 PB - American Chemical Society CY - Bethesda ER - TY - THES A1 - Gonzalez Camargo, Rodolfo T1 - Insulin resistance in cancer cachexia and metabolic syndrome BT - role of insulin activated macrophages and miRNA-21-5p N2 - The ever-increasing fat content in Western diet, combined with decreased levels of physical activity, greatly enhance the incidence of metabolic-related diseases. Cancer cachexia (CC) and Metabolic syndrome (MetS) are both multifactorial highly complex metabolism related syndromes, whose etiology is not fully understood, as the mechanisms underlying their development are not completely unveiled. Nevertheless, despite being considered “opposite sides”, MetS and CC share several common issues such as insulin resistance and low-grade inflammation. In these scenarios, tissue macrophages act as key players, due to their capacity to produce and release inflammatory mediators. One of the main features of MetS is hyperinsulinemia, which is generally associated with an attempt of the β-cell to compensate for diminished insulin sensitivity (insulin resistance). There is growing evidence that hyperinsulinemia per se may contribute to the development of insulin resistance, through the establishment of low grade inflammation in insulin responsive tissues, especially in the liver (as insulin is secreted by the pancreas into the portal circulation). The hypothesis of the present study was that insulin may itself provoke an inflammatory response culminating in diminished hepatic insulin sensitivity. To address this premise, firstly, human cell line U937 differentiated macrophages were exposed to insulin, LPS and PGE2. In these cells, insulin significantly augmented the gene expression of the pro-inflammatory mediators IL-1β, IL-8, CCL2, Oncostatin M (OSM) and microsomal prostaglandin E2 synthase (mPGES1), and of the anti-inflammatory mediator IL-10. Moreover, the synergism between insulin and LPS enhanced the induction provoked by LPS in IL-1β, IL-8, IL-6, CCL2 and TNF-α gene. When combined with PGE2, insulin enhanced the induction provoked by PGE2 in IL-1β, mPGES1 and COX2, and attenuated the inhibition induced by PGE2 in CCL2 and TNF-α gene expression contributing to an enhanced inflammatory response by both mechanisms. Supernatants of insulin-treated U937 macrophages reduced the insulin-dependent induction of glucokinase in hepatocytes by 50%. Cytokines contained in the supernatant of insulin-treated U937 macrophages also activated hepatocytes ERK1/2, resulting in inhibitory serine phosphorylation of the insulin receptor substrate. Additionally, the transcription factor STAT3 was activated by phosphorylation resulting in the induction of SOCS3, which is capable of interrupting the insulin receptor signal chain. MicroRNAs, non-coding RNAs linked to protein expression regulation, nowadays recognized as active players in the generation of several inflammatory disorders such as cancer and type II diabetes are also of interest. Considering that in cancer cachexia, patients are highly affected by insulin resistance and inflammation, control, non-cachectic and cachectic cancer patients were selected and the respective circulating levels of pro-inflammatory mediators and microRNA-21-5p, a posttranscriptional regulator of STAT3 expression, assessed and correlated. Cachectic patients circulating cytokines IL-6 and IL-8 levels were significantly higher than those of non-cachectic and controls, and the expression of microRNA-21-5p was significantly lower. Additionally, microRNA-21-5p reduced expression correlated negatively with IL-6 plasma levels. These results indicate that hyperinsulinemia per se might contribute to the low grade inflammation prevailing in MetS patients and thereby promote the development of insulin resistance particularly in the liver. Diminished MicroRNA-21-5p expression may enhance inflammation and STAT3 expression in cachectic patients, contributing to the development of insulin resistance. N2 - O teor de gordura cada vez maior na dieta ocidental, combinada com a diminuição dos níveis de atividade física têm marcadamente aumentado à incidência de doenças relacionas ao metabolismo. A caquexia associada ao câncer (CC) e a síndrome metabólica (SM) são síndromes de etiologia complexa e multifatorial, não totalmente compreendida, e com mecanismos subjacentes ao seu desenvolvimento não completamente revelados. No entanto, apesar de serem consideradas "lados opostos", a CC e a MetS apresentam várias características em comum, tais como resistência à insulina e inflamação de baixo grau, com macrófagos teciduais como importantes coadjuvantes, devido à sua capacidade de produzir e liberar mediadores inflamatórios, e microRNAs, descritos como RNAs não-codificantes ligados à regulação da expressão de proteínas e reconhecidos como participantes ativos na geração de várias doenças inflamatórias, tais como o câncer e diabetes tipo II. Uma das principais características da MetS é a hiperinsulinemia, que está geralmente associada com uma tentativa da célula β do pâncreas de compensar a diminuição da sensibilidade à insulina (resistência à insulina). Um número crescente de evidências sugere que a hiperinsulinemia “por si só”, pode contribuir com o desenvolvimento de resistência à insulina através do estabelecimento de um quadro inflamatório de baixo grau, em tecidos sensíveis a insulina, e em particular no fígado, devido ao fato da insulina ser secretada pelo pâncreas na circulação portal. A hipótese do presente estudo foi que a insulina pode induzir uma resposta inflamatória em macrófagos e culminar em diminuição da sensibilidade hepática à insulina. Para confirmar esta hipótese, primeiramente, macrófagos diferenciados da linhagem de células humanas U937 foram expostos à insulina, LPS e PGE2. Nestas células, a insulina aumentou significativamente a expressão gênica dos mediadores pró-inflamatórios IL-1β, IL- 8, CCL2, oncostatina M (OSM) e prostaglandina E2 sintase microssomal (mPGES1), e do mediador anti-inflamatório IL-10. Além disso, o sinergismo entre insulina e LPS aumentou a indução provocada por LPS nos genes da IL-1β, IL-8, IL-6, CCL2 e TNF-α. Quando combinado com PGE2, a insulina aumentou a indução provocada pela PGE2 nos genes da IL-1β, mPGES1 e COX2, e restaurou a inibição induzida pela PGE2 no gene CCL2 e TNF-α.Subsequentemente, sobrenadantes dos macrófagos U937 tratados com insulina modulou negativamente a sinalização da insulina em culturas primárias de hepatócitos de rato, como observado pela atenuação de 50% da indução dependente de insulina da enzima glicoquinase. Citocinas contidas no sobrenadante de macrófagos U937 tratados com insulina também ativaram em hepatócitos ERK1/2, resultando na fosforilação do resíduo de serina inibitório do substrato do receptor de insulina. Adicionalmente, o fator de transcrição STAT3 foi ativado por um elevado grau de fosforilação e a proteína SOCS3, capaz de interromper a via de sinalização do receptor de insulina, foi induzida. Considerando que na caquexia associada ao câncer, pacientes são altamente afetados pela resistência à insulina e inflamação, pacientes controle, não caquéticos e caquéticos foram seleccionados e os respectivos níveis circulantes de mediadores pró-inflamatórios e microRNA-21-5p, um regulador pós-transcricional da expressão de STAT3, avaliados e correlacionados. Pacientes caquéticos exibiram citocinas circulantes IL-6 e IL-8 significativamente maiores do que pacientes não caquéticos e controles, assim como a expressão de microRNA-21-5p significativamente diminuida. Além disso, a reduzida expressão de microRNA-21-5p correlaciona-se negativamente com níveis de IL-6 no plasma. Estes resultados indicam que a hiperinsulinemia pode, por si só contribuir para o desenvolvimento da inflamação de baixo grau prevalente em pacientes com excesso de peso e obesos e, assim, promover o desenvolvimento de resistência à insulina especialmente no fígado e o nível reduzido de miRNA-21-5p pode modular a inflamação e expressão de STAT3 em pacientes caquéticos, contribuindo para o desenvolvimento da resistência à insulina. N2 - Der stetig steigende Fettgehalt in westlicher Ernährung in Kombination mit reduzierter körperlicher Aktivität hat zu einem dramatischen Anstieg der Inzidenz metabolischer Erkrankungen geführt. Tumorkachexie (Cancer cachexia, CC) und Metabolisches Syndrom (MetS) sind sehr komplexe, multifaktorielle metabolische Erkrankungen, deren Ätiologie nicht vollständig verstanden ist. Die molekularen Ursachen, die zu diesen Symptomkomplexen führen, sind noch unzureichend aufgeklärt. Obwohl ihr äußeres Erscheinungsbild stark gegensätzlich ist, haben MetS und CC etliche Gemeinsamkeiten wie zum Beispiel Insulinresistenz und eine chronische unterschwellige Entzündung. Sowohl bei der Entstehung der Insulinresistenz als auch bei der chronischen Entzündung spielen Makrophagen eine Schlüsselrolle, weil sie in der Lage sind pro-inflammatorische Mediatoren zu produzieren und freizusetzen. Eine der hervorstechendsten Auffälligkeiten des MetS ist die Hyperinsulinämie, die durch den Versuch der β-Zelle, die verminderte Insulinsensitivität (Insulinresistenz) zu kompensieren, zustande kommt. Es gibt zunehmend Hinweise darauf, dass die Hyperinsulinämie selber an der Entzündungsentstehung in Insulin-abhängigen Geweben beteiligt ist und dadurch zur Entwicklung und Verstärkung der Insulinresistenz beitragen kann. Dies trifft besonders auf die Leber zu, weil hier die Insulinspiegel besonders hoch sind, da Insulin vom Pankreas direkt in den Pfortaderkeislauf gelangt. Daher wurde in dieser Arbeit die Hypothese geprüft, ob Insulin selber eine Entzündungsantwort auslösen und dadurch die hepatische Insulinsensitivität senken kann. Zu diesem Zweck wurde die humane Zelllinie U937 durch PMA-Behandlung zu Makrophagen differenziert und diese Makrophagen mit Insulin, LPS und PGE2 inkubiert. In diesen Zellen steigerte Insulin die Expression der pro-inflammatorischen Mediatoren IL-1β, IL-8, CCL2, Oncostatin M (OSM) signifikant und induzierte die mikrosomale PGE-Synthase 1 (mPGES1) ebenso wie das antiinflammatorische Cytokin IL-10. Ferner verstärkte Insulin die LPS-abhängige Induktion des IL-1β-, IL-8-, IL-6-, CCL2- und TNFα-Gens. Ebenso verstärkte Insulin die PGE2-abhängige Induktion von IL-1β, mPGES1 und COX2. Im Gegensatz dazu schwächte es die Hemmende Wirkung von PGE2 auf Expression von TNFα und CCL2 ab und trug so auf beide Weisen zu einer Verstärkung der Entzündungsantwort bei. Überstände von Insulin-behandelten U937 Makrophagen reduzierten die Insulin-abhängige Induktion der Glukokinase in Hepatocyten um 50%. Die Cytokine, die im Überstand Insulin-behandelter Makrophagen enthalten waren, aktivierten in Hepatocyten ERK1/2, was zu einer inhibitorischen Serin-Phosphorylierung der Insulin Rezeptor Substrats (IRS) führte. Zusätzlich führten die Cytokine zu einer Phosphorylierung und Aktivierung von STAT3 und einer dadurch bedingten Induktion von SOCS3, das seinerseits die Insulinrezeptor-Signalkette unterbrechen kann. MicroRNAs, nicht-codierende RNAs, die an der Regulation der Proteinexpression beteiligt sind und deren Beteiligung an der Regulation der Entzündungsantwort bei zahlreichen Erkrankungen, unter anderem Tumorerkrankungen und Typ II Diabetes gezeigt wurde, sind auch von Interesse. Unter dem Blickwinkel, dass Tumor-Kachexie Patienten sich durch eine Insulinresistenz und eine systemische Entzündung auszeichnen, wurden in nichtkachektische und tumorkachektische Patienten Plasmaspiegel von pro-inflammatorischen Mediatoren und der microRNA-21-5p bestimmt, von der bekannt ist, dass sie ein posttranskriptioneller Regulator der STAT3 Expression ist. Die Spiegel der proinflammatorischen Mediatoren und der miRNA-21-5p wurden korreliert. In kachektischen Patienten waren die Spiegel der Cytokine IL-6 und IL-8 signifikant höher, die der miRNA-21- 5p signifikant niedriger als in nicht-kachektischen Patienten. Die Plasma IL-6-Spiegel korrelierten negativ mit den miRNA21-5p Spiegeln. Insgesamt zeigen die Ergebnisse, dass eine Hyperinsulinämie selber zu der Entwicklung einer unterschwellingen Entzündung, wie sie in Patienten mit einem MetS vorherrscht, beitragen, und dadurch besonders in der Leber eine Insulinresistenz auslösen oder verstärken kann. Eine verringerte Expression der MicroRNA-21-5p kann in kachektischen Patienten die Entzündungsantwort, im Speziellen die STAT3 Expression, verstärken und dadurch zur Entwicklung einer Insulinresistenz beitragen KW - cachexia KW - metabolic syndrome KW - inflammation KW - insulin resistance KW - microRNAs KW - insulin KW - liver KW - macrophages KW - caquexia KW - síndrome metabólica KW - inflamação KW - resistência à insulina KW - microRNAs KW - insulina KW - fígado KW - macrófagos KW - Kachexie KW - metabolisches Syndrom KW - Entzündung KW - Insulinresistenz KW - MicroRNAs KW - Insulin KW - Leber KW - Makrophagen Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-100973 ER - TY - THES A1 - Luckert, Claudia T1 - Molekulare Mechanismen von hepatotoxischen Pyrrolizidinalkaloiden Y1 - 2016 ER - TY - THES A1 - Reinke, Julia T1 - The Role of Kallistatin in Energy Metabolism and Glucose Homeostasis in Mice Y1 - 2016 ER - TY - GEN A1 - Woting, Anni A1 - Blaut, Michael T1 - The intestinal microbiota in metabolic disease T2 - Nutrients N2 - Gut bacteria exert beneficial and harmful effects in metabolic diseases as deduced from the comparison of germfree and conventional mice and from fecal transplantation studies. Compositional microbial changes in diseased subjects have been linked to adiposity, type 2 diabetes and dyslipidemia. Promotion of an increased expression of intestinal nutrient transporters or a modified lipid and bile acid metabolism by the intestinal microbiota could result in an increased nutrient absorption by the host. The degradation of dietary fiber and the subsequent fermentation of monosaccharides to short-chain fatty acids (SCFA) is one of the most controversially discussed mechanisms of how gut bacteria impact host physiology. Fibers reduce the energy density of the diet, and the resulting SCFA promote intestinal gluconeogenesis, incretin formation and subsequently satiety. However, SCFA also deliver energy to the host and support liponeogenesis. Thus far, there is little knowledge on bacterial species that promote or prevent metabolic disease. Clostridium ramosum and Enterococcus cloacae were demonstrated to promote obesity in gnotobiotic mouse models, whereas bifidobacteria and Akkermansia muciniphila were associated with favorable phenotypes in conventional mice, especially when oligofructose was fed. How diet modulates the gut microbiota towards a beneficial or harmful composition needs further research. Gnotobiotic animals are a valuable tool to elucidate mechanisms underlying diet-host-microbe interactions. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 448 KW - intestinal microbiota KW - obesity KW - diabetes KW - metabolic syndrome KW - energy harvest KW - diet KW - absorption KW - bile acids KW - low-grade inflammation KW - SCFA Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-407687 ER - TY - GEN A1 - Kühn, Tilman A1 - Floegel, Anna A1 - Sookthai, Disorn A1 - Johnson, Theron A1 - Rolle-Kampczyk, Ulrike A1 - Otto, Wolfgang A1 - von Bergen, Martin A1 - Boeing, Heiner A1 - Kaaks, Rudolf T1 - Higher plasma levels of lysophosphatidylcholine 18:0 are related to a lower risk of common cancers in a prospective metabolomics study T2 - BMC medicine N2 - Background: First metabolomics studies have indicated that metabolic fingerprints from accessible tissues might be useful to better understand the etiological links between metabolism and cancer. However, there is still a lack of prospective metabolomics studies on pre-diagnostic metabolic alterations and cancer risk. Methods: Associations between pre-diagnostic levels of 120 circulating metabolites (acylcarnitines, amino acids, biogenic amines, phosphatidylcholines, sphingolipids, and hexoses) and the risks of breast, prostate, and colorectal cancer were evaluated by Cox regression analyses using data of a prospective case-cohort study including 835 incident cancer cases. Results: The median follow-up duration was 8.3 years among non-cases and 6.5 years among incident cases of cancer. Higher levels of lysophosphatidylcholines (lysoPCs), and especially lysoPC a C18:0, were consistently related to lower risks of breast, prostate, and colorectal cancer, independent of background factors. In contrast, higher levels of phosphatidylcholine PC ae C30:0 were associated with increased cancer risk. There was no heterogeneity in the observed associations by lag time between blood draw and cancer diagnosis. Conclusion: Changes in blood lipid composition precede the diagnosis of common malignancies by several years. Considering the consistency of the present results across three cancer types the observed alterations point to a global metabolic shift in phosphatidylcholine metabolism that may drive tumorigenesis. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 437 KW - metabolomics KW - epidemiology KW - breast cancer KW - prostate cancer KW - colorectal cancer Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-407258 ER - TY - JOUR A1 - Wilde, Sandra Catharina A1 - Treitz, Christian A1 - Keppler, Julia Katharina A1 - Koudelka, Tomas A1 - Palani, Kalpana A1 - Tholey, Andreas A1 - Rawel, Harshadrai Manilal A1 - Schwarz, Karin T1 - beta-Lactoglobulin as nanotransporter - Part II: Characterization of the covalent protein modification by allicin and diallyl disulfide JF - Food chemistry N2 - The whey protein beta-lactoglobulin has been proposed as a transporter for covalent bound bioactive compounds in order to enhance their stability and reduce their sensory perception. The garlic derived compounds allicin and diallyl disulfide were bound covalently to the native and heat denatured protein. The binding site and the influence of the modification on the digestibility were determined by mass spectrometric analysis of the modified beta-lactoglobulin. Further, the conformation of the modified protein was assessed by circular dichroism and dynamic light scattering. The free thiol group of Cys(121) turned out to be the major binding site. After proteolysis with trypsin at pH 7 but not with pepsin at pH 2, a limited transfer to other cysteinyl residues was observed. The covalently bound ligands did not mask any proteolytic cleavage sites of pepsin, trypsin or chymotrypsin. The modified beta-lactoglobulin showed a native like conformation, besides a moderate loosening of protein folding. The covalent binding of organosulfur compounds to beta-lactoglobulin provides a bioactive ingredient without impairing the digestibility and functional properties of the protein. (C) 2015 Elsevier Ltd. All rights reserved. KW - Beta-lactoglobulin KW - Covalent modification KW - LC-MS KW - CD, DLS KW - Thiol KW - Allicin KW - Garlic KW - Diallyl disulfide Y1 - 2016 U6 - https://doi.org/10.1016/j.foodchem.2015.11.011 SN - 0308-8146 SN - 1873-7072 VL - 197 SP - 1022 EP - 1029 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Hecht, Eva A1 - Freise, Christian A1 - von Websky, Karoline A1 - Nasser, Hamoud A1 - Kretzschmar, Nadja A1 - Stawowy, Philipp A1 - Hocher, Berthold A1 - Querfeld, Uwe T1 - The matrix metalloproteinases 2 and 9 initiate uraemic vascular calcifications JF - Nephrology, dialysis, transplantation N2 - The matrix metalloproteinases (MMP) MMP-2 and MMP-9 are physiological regulators of vascular remodelling. Their dysregulation could contribute to vascular calcification. We examined the role of the MMP-2 and MMP-9 in uraemic vascular calcification in vivo and in vitro. The impact of pharmacological MMP inhibition on the development of media calcifications was explored in an aggressive animal model of uraemic calcification. In addition, the selective effects of addition and inhibition, respectively, of MMP-2 and MMP-9 on calcium-/phosphate-induced calcifications were studied in a murine cell line of vascular smooth muscle cells (VSMCs). High-dose calcitriol treatment of uraemic rats given a high phosphate diet induced massive calcifications, apoptosis and increased gene expressions of MMP-2, MMP-9 and of osteogenic transcription factors and proteins in aortic VSMC. The MMP inhibitor doxycycline prevented the VSMC transdifferentiation to osteoblastic cells, suppressed transcription of mediators of matrix remodelling and almost completely blocked aortic calcifications while further increasing apoptosis. Similarly, specific inhibitors of either MMP-2 or -9, or of both gelatinases (Ro28-2653) and a selective knockdown of MMP-2/-9 mRNA expression blocked calcification of murine VSMC induced by calcification medium (CM). In contrast to MMP inhibition, recombinant MMP-2 or MMP-9 enhanced CM-induced calcifications and the secretion of gelatinases. These data indicate that both gelatinases provide essential signals for phenotypic VSMC conversion, matrix remodelling and the initiation of vascular calcification. Their inhibition seems a promising strategy in the prevention of vascular calcifications. KW - chronic kidney disease KW - matrix metalloproteinases KW - vascular calcification KW - vascular smooth muscle cells Y1 - 2016 U6 - https://doi.org/10.1093/ndt/gfv321 SN - 0931-0509 SN - 1460-2385 VL - 31 SP - 789 EP - 797 PB - Oxford Univ. Press CY - Oxford ER - TY - GEN A1 - Bijleveld, Catrien A1 - Zoutewelle-Terovan, Mioara A1 - Huschek, Doreen A1 - Liefbroer, Aart C. T1 - Criminal careers and demographic outcomes: An introduction to the special issue T2 - Advances in life course research Y1 - 2016 U6 - https://doi.org/10.1016/j.alcr.2016.05.001 SN - 1569-4909 SN - 1879-6974 VL - 28 SP - 1 EP - 5 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Bijleveld, Catrien A1 - Huschek, Doreen A1 - Liefbroer, Aart C. T1 - Parental criminality and entry into parenthood among sons and daughters JF - Advances in life course research N2 - In this article, we examined to what extent parental offending influences the timing of entry into parenthood of children. Based on a literature review, we hypothesized that children of delinquent parents would be more likely to enter into parenthood at a relatively young age, and that part of that association could be explained by differences between children of delinquent and non-delinquent parents in the timing of entry into marriage and in their own delinquent behaviour. Using data from a five-generation study of high risk families in the Netherlands, we found that parental delinquency increases the chance of early childbearing among daughters, but not among sons. Among sons, parental delinquency increased son's delinquency, suggesting that parental delinquency has different consequences for the life courses of their sons and daughters. KW - Intergenerational effects KW - Offending KW - Parenthood KW - Demographic transitions KW - Delinquency Y1 - 2016 U6 - https://doi.org/10.1016/j.alcr.2016.03.006 SN - 1569-4909 SN - 1879-6974 VL - 28 SP - 81 EP - 90 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Sievers, Steven A1 - Rawel, Harshadrai Manilal A1 - Ringel, Karl Peter A1 - Niggemann, Bodo A1 - Beyer, Kirsten T1 - Wheat protein recognition pattern in tolerant and allergic children JF - Pediatric Allergy and Immunology N2 - BackgroundWheat is one of the most common food allergens in early childhood. In contrast to other food allergies, wheat-specific IgE correlates badly with clinical symptoms and relevant components have been identified mostly for wheat-depended exercise-induced anaphylaxis. Moreover, a high percentage of patients present with immediate type symptoms but wheat-specific IgE cannot be detected with commercial available systems. ObjectiveWe addressed the question whether the IgE recognition pattern between wheat allergic (WA) and clinically tolerant (WT) children differs in order to identify individual proteins useful for component-resolved diagnostics. MethodsSera of 106 children with suspected wheat allergy, of whom 44 children had clinical relevant wheat allergy and 62 were tolerant upon oral food challenge, were analyzed for wheat-specific IgE using the ImmunoCap system as well as immunoblots against water and salt soluble, and water-insoluble protein fractions. 40 randomly selected sera were analyzed for specific IgE to 5-gliadin. ResultsSixty-three percent of the WT and 86% of the WA children were sensitized to wheat with >0.35 kU(A)/l in ImmunoCAP analysis. We could confirm the role of -, ss-, -, and -gliadins, and LMW glutenin subunits as major allergens and found also IgE binding to a broad spectrum of water- and salt-soluble protein bands. It is of great importance that wheat allergic and tolerant patients showed IgE binding to the same protein bands. WT and WA did not significantly differ in levels of 5-gliadin-specific IgE. Conclusions & Clinical RelevanceChildren with challenge proven clinical relevant food allergy and tolerant ones had a similar spectrum of IgE binding to the same protein bands. These findings imply that component-resolved diagnostics might not be helpful in the diagnostic work-up of wheat allergy. KW - wheat KW - IgE KW - 5-gliadin KW - protein pattern KW - immunoblot Y1 - 2016 U6 - https://doi.org/10.1111/pai.12502 SN - 0905-6157 SN - 1399-3038 VL - 27 SP - 147 EP - 155 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Schmiedchen, Bettina A1 - Longardt, Ann Carolin A1 - Loui, Andrea A1 - Buehrer, Christoph A1 - Raila, Jens A1 - Schweigert, Florian J. T1 - Effect of vitamin A supplementation on the urinary retinol excretion in very low birth weight infants JF - European journal of pediatrics : official organ of the Belgian Pediatric Association N2 - Despite high-dose vitamin A supplementation of very low birth weight infants (VLBW, <1500 g), their vitamin A status does not improve substantially. Unknown is the impact of urinary retinol excretion on the serum retinol concentration in these infants. Therefore, the effect of high-dose vitamin A supplementation on the urinary vitamin A excretion in VLBW infants was investigated. Sixty-three VLBW infants were treated with vitamin A (5000 IU intramuscular, 3 times/week for 4 weeks); 38 untreated infants were classified as control group. On days 3 and 28 of life, retinol, retinol-binding protein 4 (RBP4), glomerular filtration rate, proteinuria, and Tamm-Horsfall protein were quantified in urine. On day 3 of life, substantial retinol and RBP4 losses were found in both groups, which significantly decreased until day 28. Notwithstanding, the retinol excretion was higher (P<0.01) under vitamin A supplementation as compared to infants of the control group. On day 28 of life, the urinary retinol concentrations were predictive for serum retinol concentrations in the vitamin A treated (P<0.01), but not in the control group (P=0.570). Conclusion: High urinary retinol excretion may limit the vitamin A supplementation efficacy in VLBW infants. Advanced age and thus postnatal kidney maturation seems to be an important contributor in the prevention of urinary retinol losses. KW - Vitamin A supplementation KW - RBP4 KW - Very low birth weight infant KW - Urine excretion Y1 - 2016 U6 - https://doi.org/10.1007/s00431-015-2647-9 SN - 0340-6199 SN - 1432-1076 VL - 175 SP - 365 EP - 372 PB - Springer CY - New York ER - TY - JOUR A1 - Rosenkranz, Eva A1 - Maywald, Martina A1 - Hilgers, Ralf-Dieter A1 - Brieger, Anne A1 - Clarner, Tim A1 - Kipp, Markus A1 - Pluemaekers, Birgit A1 - Meyer, Sören A1 - Schwerdtle, Tanja A1 - Rink, Lothar T1 - Induction of regulatory T cells in Th1-/Th17-driven experimental autoimmune encephalomyelitis by zinc administration JF - The journal of nutritional biochemistry N2 - The essential trace element zinc is indispensable for proper immune function as zinc deficiency accompanies immune defects and dysregulations like allergies, autoimmunity and an increased presence of transplant rejection. This point to the importance of the physiological and dietary control of zinc levels for a functioning immune system. This study investigates the capacity of zinc to induce immune tolerance. The beneficial impact of physiological zinc supplementation of 6 mu g/day (0.3 mg/kg body weight) or 30 mu g/day (1.5 mg/kg body weight) on murine experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis with a Th1/Th17 (Th, T helper) cell-dominated immunopathogenesis, was analyzed. Zinc administration diminished EAE scores in C57BL/6 mice in vivo (P<.05), reduced Th17 ROR gamma T+ cells (P<.05) and significantly increased inducible iTreg cells (P<.05). While Th17 cells decreased systemically, iTreg cells accumulated in the central nervous system. Cumulatively, zinc supplementation seems to be capable to induce tolerance in unwanted immune reactions by increasing iTreg cells. This makes zinc a promising future tool for treating autoimmune diseases without suppressing the immune system. (C) 2015 Elsevier Inc. All rights reserved. KW - Zinc KW - Regulatory T cells (Treg) KW - Foxp3 KW - Mixed lymphocyte culture (MLC) KW - Experimental autoimmune encephalomyelitis (EAE) KW - Th17 Y1 - 2016 U6 - https://doi.org/10.1016/j.jnutbio.2015.11.010 SN - 0955-2863 SN - 1873-4847 VL - 29 SP - 116 EP - 123 PB - Elsevier CY - New York ER - TY - JOUR A1 - Passlack, Nadine A1 - Schmiedchen, Bettina A1 - Raila, Jens A1 - Schweigert, Florian J. A1 - Stumpff, Friederike A1 - Kohn, Barbara A1 - Neumann, Konrad A1 - Zentek, Juergen T1 - Impact of Increasing Dietary Calcium Levels on Calcium Excretion and Vitamin D Metabolites in the Blood of Healthy Adult Cats JF - PLoS one N2 - Background Dietary calcium (Ca) concentrations might affect regulatory pathways within the Ca and vitamin D metabolism and consequently excretory mechanisms. Considering large variations in Ca concentrations of feline diets, the physiological impact on Ca homeostasis has not been evaluated to date. In the present study, diets with increasing concentrations of dicalcium phosphate were offered to ten healthy adult cats (Ca/phosphorus (P): 6.23/6.02, 7.77/7.56, 15.0/12.7, 19.0/17.3, 22.2/19.9, 24.3/21.6 g/kg dry matter). Each feeding period was divided into a 10-day adaptation and an 8-day sampling period in order to collect urine and faeces. On the last day of each feeding period, blood samples were taken. Results Urinary Ca concentrations remained unaffected, but faecal Ca concentrations increased (P < 0.001) with increasing dietary Ca levels. No effect on whole and intact parathyroid hormone levels, fibroblast growth factor 23 and calcitriol concentrations in the blood of the cats were observed. However, the calcitriol precursors 25(OH)D-2 and 25(OH)D-3, which are considered the most useful indicators for the vitamin D status, decreased with higher dietary Ca levels (P = 0.013 and P = 0.033). Increasing dietary levels of dicalcium phosphate revealed an acidifying effect on urinary fasting pH (6.02) and postprandial pH (6.01) (P < 0.001), possibly mediated by an increase of urinary phosphorus (P) concentrations (P < 0.001). Conclusions In conclusion, calcitriol precursors were linearly affected by increasing dietary Ca concentrations. The increase in faecal Ca excretion indicates that Ca homeostasis of cats is mainly regulated in the intestine and not by the kidneys. Long-term studies should investigate the physiological relevance of the acidifying effect observed when feeding diets high in Ca and P. Y1 - 2016 U6 - https://doi.org/10.1371/journal.pone.0149190 SN - 1932-6203 VL - 11 SP - 47 EP - 67 PB - PLoS CY - San Fransisco ER - TY - JOUR A1 - Sandmann, Michael A1 - Garz, Andreas A1 - Menzel, Ralf T1 - Physiological response of two different Chlamydomonas reinhardtii strains to light-dark rhythms JF - Botany N2 - Cells of a cell-wall deficient line (cw15-type) of Chlamydomonas reinhardtii and of the corresponding wild type were grown during repetitive light-dark cycles. In a direct comparison, both lines showed approximately the same relative biomass increase during light phase but the cw-line produced significantly more, and smaller, daughter cells. Throughout the light period the average cellular starch content, the cellular chlorophyll content, the cellular rate of dark respiration, and the cellular rate of photosynthesis of the cw-line was lower. Despite this, several non-cell volume related parameters like the development of starch content per cell volume were clearly different over time between the strains. Additionally, the chlorophyll-based photosynthesis rates were 2-fold higher in the mutant than in the wild-type cells, and the ratio of chlorophyll a to chlorophyll b as well as the light-saturation index were also consistently higher in the mutant cells. Differences in the starch content were also confirmed by single cell analyses using a sensitive SHG-based microscopy approach. In summary, the cw15-type mutant deviates from its genetic background in the entire cell physiology. Both lines should be used in further studies in comparative systems biology with focus on the detailed relation between cell volume increase, photosynthesis, starch metabolism, and daughter cell productivity. KW - cell wall deficient mutant KW - diurnal rhythm KW - nonlinear microscopy KW - photosynthesis KW - single-cell analysis Y1 - 2016 U6 - https://doi.org/10.1139/cjb-2015-0144 SN - 1916-2790 SN - 1916-2804 VL - 94 SP - 53 EP - 64 PB - NRC Research Press CY - Ottawa ER - TY - JOUR A1 - Li, Jian A1 - Lu, Yong Ping A1 - Reichetzeder, Christoph A1 - Kalk, Philipp A1 - Kleuser, Burkhard A1 - Adamski, Jerzy A1 - Hocher, Berthold T1 - Maternal PCaaC38:6 is Associated With Preterm Birth - a Risk Factor for Early and Late Adverse Outcome of the Offspring JF - Journal of European public policy N2 - Background/Aims: Preterm birth (PTB) and low birth weight (LBW) significantly influence mortality and morbidity of the offspring in early life and also have long-term consequences in later life. A better understanding of the molecular mechanisms of preterm birth could provide new insights regarding putative preventive strategies. Metabolomics provides a powerful analytic tool to readout complex interactions between genetics, environment and health and may serve to identify relevant biomarkers. In this study, the association between 163 targeted maternal blood metabolites and gestational age was investigated in order to find candidate biomarkers for PTB. Methods: Five hundred twenty-three women were included into this observational study. Maternal blood was obtained before delivery. The concentration of 163 maternal serum metabolites was measured by flow injection tandem mass spectrometry. To find putative biomarkers for preterm birth, a three-step analysis was designed: bivariate correlation analysis followed by multivariable regression analysis and a comparison of mean values among gestational age groups. Results: Bivariate correlation analysis showed that 2 acylcarnitines (C16:2, C2), 1 amino acids (xLeu), 8 diacyl-PCs (PCaaC36:4, PCaaC38:4, PCaaC38:5, PCaaC38:6, PCaaC40:4, PCaaC40:5, PCaaC40:6, PCaaC42:4), and 1 Acylalkyl-PCs (PCaeC40:5) were inversely correlated with gestational age. Multivariable regression analysis confounded for PTB history, maternal body mass index (BMI) before pregnancy, systolic blood pressure at the third trimester, and maternal body weight at the third trimester, showed that the diacyl-PC PCaaC38:6 was the only metabolite inversely correlated with gestational age. Conclusions: Maternal blood concentrations of PCaaC38:6 are independently associated with gestational age. (C) 2016 The Author(s) Published by S. Karger AG, Basel KW - Metabolomics KW - PCaaC38:6 KW - Biomarker KW - Preterm birth Y1 - 2016 U6 - https://doi.org/10.1159/000443428 SN - 1420-4096 SN - 1423-0143 VL - 41 SP - 250 EP - 257 PB - Karger CY - Basel ER - TY - JOUR A1 - Ebert, Franziska A1 - Meyer, Sören A1 - Leffers, Larissa A1 - Raber, Georg A1 - Francesconi, Kevin A. A1 - Schwerdtle, Tanja T1 - Toxicological characterisation of a thio-arsenosugar-glycerol in human cells JF - Journal of trace elements in medicine and biology N2 - Arsenosugars are water-soluble arsenic species predominant in marine algae and other seafood including mussels and oysters. They typically occur at levels ranging from 2 to 50 mg arsenic/kg dry weight. Most of the arsenosugars contain arsenic as a dimethylarsinoyl group (Me2As(O)-), commonly referred to as the oxo forms, but thio analogues have also been identified in marine organisms and as metabolic products of oxo-arsenosugars. So far, no data regarding toxicity and toxicokinetics of thio-arsenosugars are available. This in vitro-based study indicates that thio-dimethylarsenosugar-glycerol exerts neither pronounced cytotoxicity nor genotoxicity even though this arsenical was bioavailable to human hepatic (HepG2) and urothelial (UROtsa) cells. Experiments with the Caco-2 intestinal barrier model mimicking human absorption indicate for the thio-arsenosugar-glycerol higher intestinal bioavailability as compared to the oxo-arsenosugars. Nevertheless, absorption estimates were much lower in comparison to other arsenicals including arsenite and arsenic-containing hydrocarbons. Arsenic speciation in cell lysates revealed that HepG2 cells are able to metabolise the thio-arsenosugar-glycerol to some extent to dimethylarsinic acid (DMA). These first in vitro data cannot fully exclude risks to human health related to the presence of thio-arsenosugars in food. (C) 2016 Elsevier GmbH. All rights reserved. KW - Arsenic KW - Thio-arsenosugar-glycerol KW - Toxicity KW - Toxicokinetics KW - Genotoxicity KW - Metabolism Y1 - 2016 U6 - https://doi.org/10.1016/j.jtemb.2016.04.013 SN - 0946-672X VL - 38 SP - 150 EP - 156 PB - Springer Publishing Company CY - Jena ER - TY - JOUR A1 - Ebert, Franziska A1 - Thomann, Marlies A1 - Witt, Barbara A1 - Müller, Sandra Marie A1 - Meyer, Sören A1 - Weber, Till A1 - Christmann, Markus A1 - Schwerdtle, Tanja T1 - Evaluating long-term cellular effects of the arsenic species thio-DMA(V): qPCR-based gene expression as screening tool JF - Journal of trace elements in medicine and biology N2 - Thio-dimethylarsinic acid (thio-DMA(V)) is a human urinary metabolite of the class 1 human carcinogen inorganic arsenic as well as of arsenosugars. Thio-DMA(V) exerts strong cellular toxicity, whereas its toxic modes of action are not fully understood. For the first time, this study characterises the impact of a long-term (21 days) in vitro incubation of thio-DMA(V) on the expression of selected genes related to cell death, stress response, epigenetics and DNA repair. The observed upregulation of DNMT1 might be a cellular compensation to counterregulate the in a very recent study observed massive global DNA hypomethylation after chronic thio-DMAv incubation. Moreover, our data suggest that chronic exposure towards subcytotoxic, pico- to nanomolar concentrations of thio-DMA(V) causes a stress response in human urothelial cells. The upregulation of genes encoding for proteins of DNA repair (Apex1,Lig1, XRCC1,DDB2, XPG, ATR) as well as damage response (GADD45A, GADD45G, Trp53) indicate a potential genotoxic risk emanating from thio-DMA(V) after long-term incubation. (C) 2016 Elsevier GmbH. All rights reserved. KW - Thio-dimethylarsinic acid KW - Long-term cellular toxicity KW - qPCR-based gene expression screening KW - GADD45A and GADD45G KW - DNMT1 KW - Cellular damage response Y1 - 2016 U6 - https://doi.org/10.1016/j.jtemb.2016.06.004 SN - 0946-672X VL - 37 SP - 78 EP - 84 PB - Yokohama Publishers CY - Jena ER - TY - JOUR A1 - Forssmann, Wolf-Georg A1 - Tillmann, Hanns-Christian A1 - Hock, Dieter A1 - Forssmann, Kristin A1 - Bernasconi, Corrado A1 - Forssmann, Ulf A1 - Richter, Rudolf A1 - Hocher, Berthold A1 - Pfuetzner, Andreas T1 - Pharmacokinetic and Pharmacodynamic Characteristics of Subcutaneously Applied PTH-1-37 JF - German politics N2 - Background/Aims: Parathyroid hormone (PTH) derivatives exert pronounced renal and osteoanabolic properties when given intermittently. The current study was performed to assess the pharmacokinetic and pharmacodynamic properties as well as safety of subcutaneously applied PTH-1-37 after repeated dosing in healthy subjects. Methods: This randomized, double-blind, dose-escalating, placebo and active comparator controlled study was conducted in 33 healthy postmenopausal women. Subjects were allocated to one of five treatment options: 10, 20, or 40 mu g PTH-1-37, 20 mu g PTH-1-34 or placebo, administered as once daily subcutaneous doses for three days. Plasma drug concentrations and serum levels of endogenous PTH-1-84, and calcium as markers of biological activity were monitored during the treatment. Results: PTH was absorbed rapidly from the subcutaneous tissue with a median t(max) of 30 minutes for 20 and 40 mu g of PTH-1-37. t(max) was 45 minutes for 20 mu g PTH-1-34. Elimination half-lives were estimated as 76 +/- 34 min and 70 +/- 13 min for 20 mu g and 40 mu g PTH-1-37 (mean +/- SD), and 78 +/- 34 for 20 mu g PTH-1-34. Both PTH fragments (PTH-1-37 and PTH-1-34) increased serum calcium. For PTH-1-37 the effect on serum calcium was dose-dependent. Suppression of endogenous PTH-1-84 was seen after the application of both PTH-1-37 and PTH-1-34. During the study period, the subjects experienced no unexpected or serious adverse events. Conclusions: PTH-1-37 is rapidly absorbed after s.c. injection, has a short plasma elimination half-life, and does not accumulate during multiple dosing. Biological activity was demonstrated by rising serum calcium and decreasing endogenous PTH-1-84 in blood plasma. The study drugs were well tolerated and safe. Our investigation presents data that PTH-1-37 is an excellent drug candidate for intervening with syndromes of dysregulation of calcium metabolism. (C) 2016 The Author(s) Published by S. Karger AG, Basel KW - Pharmacokinetics Y1 - 2016 U6 - https://doi.org/10.1159/000443453 SN - 1420-4096 SN - 1423-0143 VL - 41 SP - 507 EP - 518 PB - Karger CY - Basel ER - TY - JOUR A1 - Reichetzeder, Christoph A1 - Putra, Sulistyo Emantoko Dwi A1 - Li, Jian A1 - Hocher, Berthold T1 - Developmental Origins of Disease - Crisis Precipitates Change JF - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry and pharmacology N2 - The concept of developmental origins of diseases has gained a huge interest in recent years and is a constantly emerging scientific field. First observations hereof originated from epidemiological studies, linking impaired birth outcomes to adult chronic, noncommunicable disease. By now there is a considerable amount of both epidemiological and experimental evidence highlighting the impact of early life events on later life disease susceptibility. Albeit far from being completely understood, more recent studies managed to elucidate underlying mechanisms, with epigenetics having become almost synonymous with developmental programming. The aim of this review was to give a comprehensive overview of various aspects and mechanisms of developmental origins of diseases. Starting from initial research foci mainly centered on a nutritionally impaired intrauterine environment, more recent findings such as postnatal nutrition, preterm birth, paternal programming and putative interventional approaches are summarized. The review outlines general underlying mechanisms and particularly discusses mechanistic explanations for sexual dimorphism in developmental programming. Furthermore, novel hypotheses are presented emphasizing a non-mendelian impact of parental genes on the offspring's phenotype. KW - Nutrition KW - Thrifty phenotype KW - Developmental programming KW - Paternal, maternal, sex differences KW - Epigenetics Y1 - 2016 U6 - https://doi.org/10.1159/000447801 SN - 1015-8987 SN - 1421-9778 VL - 39 SP - 919 EP - 938 PB - Karger CY - Basel ER - TY - JOUR A1 - Ahlberg, Sebastian A1 - Rancan, Fiorenza A1 - Epple, Matthias A1 - Loza, Kateryna A1 - Höppe, David A1 - Lademann, Jürgen A1 - Vogt, Annika A1 - Kleuser, Burkhard A1 - Gerecke, Christian A1 - Meinke, Martina C. T1 - Comparison of different methods to study effects of silver nanoparticles on the pro- and antioxidant status of human keratinocytes and fibroblasts JF - Methods : focusing on rapidly developing techniques KW - Oxidative stress KW - Dichlorofluorescein assay KW - Electron paramagnetic resonance spectroscopy KW - HaCaT cells KW - Glutathione KW - Free radicals Y1 - 2016 U6 - https://doi.org/10.1016/j.ymeth.2016.05.015 SN - 1046-2023 SN - 1095-9130 VL - 109 SP - 55 EP - 63 PB - Elsevier CY - San Diego ER - TY - GEN A1 - Schernthaner, G. A1 - Groop, P. A1 - Cooper, M. A1 - Perkovic, V A1 - Hocher, Berthold A1 - Kanasaki, K. A1 - Sharma, K. A1 - Stanton, R. A1 - Toto, R. A1 - Cescutti, Jessica A1 - Gordat, M. A1 - Meinicke, T. A1 - Koitka-Weber, A. A1 - Woerle, H. A1 - Eynatten, M. T1 - EFFECTS OF LINAGLIPTIN ON GLYCAEMIC CONTROL AND ALBUMINURIA IN TYPE 2 DIABETES - THE MARLINA-T2D (TM) TRIAL T2 - Nephrology Y1 - 2016 U6 - https://doi.org/10.1111/nep.12887 SN - 1320-5358 SN - 1440-1797 VL - 21 SP - 60 EP - 60 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Huschek, Gerd A1 - Boenick, Josephine A1 - Loewenstein, Yvonne A1 - Sievers, Steven A1 - Rawel, Harshadrai Manilal T1 - Quantification of allergenic plant traces in baked products by targeted proteomics using isotope marked peptides JF - LWT - food science and technology : an official journal of the Swiss Society of Food Science and Technology (SGLWT/SOSSTA) and the International Union of Food Science and Technology (IUFoST) N2 - The right choice of analytical methods for plant allergen quantification is a deciding factor for the correct assessment and labeling of allergens in processed food in view of consumer protection. The aim of the present study was to develop a validated target peptide multi-method by LC/MS/MS providing high specificity and sensitivity for plant allergen protein detection, plant identification in vegan or vegetarian products using peptide markers for quantification. The methodical concept considers the selection of target peptides of thermostable allergenic plant proteins (Gly m6 soy, Ses i6 sesame and (beta-conglutin from white lupine) by data base research, BLAST and in silico digestion using Skyline software. Different allergenic concentration levels of these proteins were integrated into our own reference bakery products and quantified with. synthesized isotopically labeled peptides after in-solution digestion using LC/MS/MS. Recovery rates within the range of 70-113% and LOQ of 10 ppm-50 ppm (mg allergenic food/kg) could be determined. The results are independent of thermal processing applied during baking and of epitope binding site for the tested allergens. (C) 2016 Elsevier Ltd. All rights reserved. KW - Allergenic food KW - Plant allergen (soy, sesame, lupine) KW - LC/MS/MS; Quantification of allergenic plant traces KW - Food labeling Y1 - 2016 U6 - https://doi.org/10.1016/j.lwt.2016.07.057 SN - 0023-6438 SN - 1096-1127 VL - 74 SP - 286 EP - 293 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Hönzke, Stefan A1 - Gerecke, Christian A1 - Elpelt, Anja A1 - Zhang, Nan A1 - Unbehauen, Michael A1 - Kral, Vivian A1 - Fleige, Emanuel A1 - Paulus, Florian A1 - Haag, Rainer A1 - Schäfer-Korting, Monika A1 - Kleuser, Burkhard A1 - Hedtrich, Sarah T1 - Tailored dendritic core-multishell nanocarriers for efficient dermal drug delivery: A systematic top-down approach from synthesis to preclinical testing JF - Journal of controlled release N2 - Drug loaded dendritic core-multishell (CMS) nanocarriers are of especial interest for the treatment of skin diseases, owing to their striking dermal delivery efficiencies following topical applications. CMS nanocarriers are composed of a polyglycerol core, connected by amide-bonds to an inner alkyl shell and an outer methoxy poly(ethylene glycol) shell. Since topically applied nanocarriers are subjected to biodegradation, the application of conventional amide-based CMS nanocarriers (10-A-18-350) has been limited by the potential production of toxic polyglycerol amines. To circumvent this issue, three tailored ester-based CMS nanocarriers (10-E-12-350, 10-E-15-350, 10-E-18-350) of varying inner alkyl chain length were synthesized and comprehensively characterized in terms of particle size, drug loading, biodegradation and dermal drug delivery efficiency. Dexamethasone (DXM), a potent drug widely used for the treatment of inflammatory skin diseases, was chosen as a therapeutically relevant test compound for the present study. Ester-and amide-based CMS nanocarriers delivered DXM more efficiently into human skin than a commercially available DXM cream. Subsequent in vitro and in vivo toxicity studies identified CMS (10-E-15-350) as the most biocompatible carrier system. The anti-inflammatory potency of DXM-loaded CMS (10-E-15-350) nanocarriers was assessed in TNF alpha supplemented skin models, where a significant reduction of the pro-inflammatory cytokine IL-8 was seen, with markedly greater efficacy than commercial DXM cream. In summary, we report the rational design and characterization of tailored, biodegradable, ester-based CMS nanocarriers, and their subsequent stepwise screening for biocompatibility, dermal delivery efficiency and therapeutic efficacy in a top-down approach yielding the best carrier system for topical applications. (C) 2016 Elsevier B.V. All rights reserved. KW - Dendritic core-multishell nanocarriers KW - Biocompatibility KW - Dexamethasone KW - Inflammatory skin disease KW - Dermal drug delivery KW - Skin model Y1 - 2016 U6 - https://doi.org/10.1016/j.jconrel.2016.06.030 SN - 0168-3659 SN - 1873-4995 VL - 242 SP - 50 EP - 63 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Sahle, Fitsum Feleke A1 - Balzus, Benjamin A1 - Gerecke, Christian A1 - Kleuser, Burkhard A1 - Bodmeier, Roland T1 - Formulation and in vitro evaluation of polymeric enteric nanoparticles as dermal carriers with pH-dependent targeting potential JF - European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences, EUFEPS N2 - pH-sensitive nanoparticles which release in a controlled fashion on the skin or dissolve in the hair follicle could significantly improve treatment effectiveness and make transfollicular drug delivery a success. Dexamethasone-loaded Eudragit L 100 nanoparticles were prepared by nanoprecipitation from an organic drug-polymer solution. Their toxicity potential was assessed using isolated human fibroblasts. pH-dependent swelling and erosion kinetics of the nanoparticles were investigated by dynamic light scattering and viscosity measurements and its effect on drug release was assessed in vitro with Franz diffusion cells. Stable, 100-550 nm-sized dexamethasone-loaded Eudragit L 100 nanoparticles with drug loading capacity and entrapment efficiency as high as 83% and 85%, respectively, were obtained by using polyvinyl alcohol as a stabilizer and ethanol as organic solvent The nanoparticles showed little or no toxicity on isolated normal human fibroblasts. Dexamethasone existed in the nanoparticles as solid solution or in amorphous form. The nanoparticles underwent extensive swelling and slow drug release in media with a low buffer capacity (as low as 10 mM) and a higher pH or at a pH close to the dissolution pH of the polymer (pH 6) and a higher buffer capacity. In 40 mM buffer and above pH 6.8, the nanoparticles eroded fast or dissolved completely and thus released the drug rapidly. pH-sensitive nanoparticles which potentially release in a controlled manner on the stratum corneum but dissolve in the hair follicle could be prepared. (C) 2016 Elsevier B.V. All rights reserved. KW - Dexamethasone KW - Enteric polymer KW - Eudragit L 100 KW - pH-sensitive nanoparticles KW - Skin nanocarrier KW - Erosion kinetics Y1 - 2016 U6 - https://doi.org/10.1016/j.ejps.2016.07.004 SN - 0928-0987 SN - 1879-0720 VL - 92 SP - 98 EP - 109 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Islam, Khan Shaiful A1 - Khalil, Mahmoud A1 - Männer, K. A1 - Raila, Jens A1 - Rawel, Harshadrai Manilal A1 - Zentek, J. A1 - Schweigert, Florian J. T1 - Effect of dietary alpha-tocopherol on the bioavailability of lutein in laying hen JF - Journal of animal physiology and animal nutrition N2 - Lutein and its isomer zeaxanthin have gained considerable interest as possible nutritional ingredient in the prevention of age-related macular degeneration (AMD) in humans. Egg yolk is a rich source of these carotenoids. As an oxidative sensitive component, antioxidants such as -tocopherol (T) might contribute to an improved accumulation in egg yolk. To test this, chickens were fed lutein esters (LE) with and without -tocopherol as an antioxidant. After depletion on a wheat-soya bean-based lutein-poor diet for 21days, laying hens (n=42) were equally divided into three groups and fed the following diets for 21days: control (basal diet), a LE group (40mg LE/kg feed) and LE+T group (40mg LE plus 100mg T/kg feed). Eggs and blood were collected periodically. Carotenoids and -tocopherol in yolk and blood plasma were determined by HPLC. Egg yolk was also analysed for total carotenoids using a one-step spectrophotometric method (iCheck(())). Lutein, zeaxanthin, -tocopherol and total carotenoids in egg yolk were highest after 14days of feeding and decreased slightly afterwards. At the end of the trial, eggs of LE+T group contained higher amount of lutein (13.72), zeaxanthin (0.65), -tocopherol (297.40) and total carotenoids (21.6) compared to the LE group (10.96, 0.55, 205.20 and 18.0mg/kg, respectively, p<0.05). Blood plasma values of LE+T group contain higher lutein (1.3), zeaxanthin (0.06) and tocopherol (20.1) compared to LE group (1.02, 0.04 and 14.90mg/l, respectively, p<0.05). In conclusion, dietary -tocopherol enhances bioavailability of lutein reflecting higher content in egg yolk and blood plasma. Improved bioavailability might be due to increased absorption of lutein in the presence of tocopherol and/or a greater stability of lutein/zeaxanthin due to the presence of -tocopherol as an antioxidant. KW - carotenoids KW - tocopherol KW - egg yolk KW - bioavailability KW - HPLC KW - iCheck Y1 - 2016 U6 - https://doi.org/10.1111/jpn.12464 SN - 0931-2439 SN - 1439-0396 VL - 100 SP - 868 EP - 875 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Niehoff, Ann-Christin A1 - Schulz, Jacqueline A1 - Soltwisch, Jens A1 - Meyer, Soren A1 - Kettling, Hans A1 - Sperling, Michael A1 - Jeibmann, Astrid A1 - Dreisewerd, Klaus A1 - Francesconi, Kevin A. A1 - Schwerdtle, Tanja A1 - Karst, Uwe T1 - Imaging by Elemental and Molecular Mass Spectrometry Reveals the Uptake of an Arsenolipid in the Brain of Drosophila melanogaster JF - Analytical chemistry N2 - Arsenic-containing lipids (arsenolipids) are natural products of marine organisms such as fish, invertebrates, and algae, many of which are important seafoods. A major group of arsenolipids, namely, the arsenic-containing hydrocarbons (AsHC), have recently been shown to be cytotoxic to human liver and bladder cells, a result that has stimulated interest in the chemistry and toxicology of these compounds. In this study, elemental laser ablation-inductively coupled plasma mass spectrometry (LA-ICPMS) and molecular matrix-assisted laser desorption/ionization (MALDI-)MS were used to image and quantify the uptake of an AsHC in the model organism Drosophila melanogaster. Using these two complementary methods, both an enrichment of arsenic and the presence of the AsHC in the brain were revealed, indicating that the intact arsenolipid had crossed the blood-brain barrier. Simultaneous acquisition of quantitative elemental concentrations and molecular distributions could allow new insight into organ-specific enrichment and possible transportation processes of arsenic-containing bioactive compounds in living organisms. Y1 - 2016 U6 - https://doi.org/10.1021/acs.analchem.6b00333 SN - 0003-2700 SN - 1520-6882 VL - 88 SP - 5258 EP - 5263 PB - American Chemical Society CY - Washington ER - TY - JOUR A1 - Tsuprykov, Oleg A1 - Ando, Ryotaro A1 - Reichetzeder, Christoph A1 - von Websky, Karoline A1 - Antonenko, Viktoriia A1 - Sharkovska, Yuliya A1 - Chaykovska, Lyubov A1 - Rahnenfuehrer, Jan A1 - Hasan, Ahmed Abdallah Abdalrahman Mohamed A1 - Tammen, Harald A1 - Alter, Markus L. A1 - Klein, Thomas A1 - Ueda, Seiji A1 - Yamagishi, Sho-ichi A1 - Okuda, Seiya A1 - Hocher, Berthold T1 - The dipeptidyl peptidase inhibitor linagliptin and the angiotensin II receptor blocker telmisartan show renal benefit by different pathways in rats with 5/6 nephrectomy JF - Kidney international : official journal of the International Society of Nephrology N2 - Dipeptidyl peptidase (DPP)-4 inhibitors delay chronic kidney disease (CKD) progression in experimental diabetic nephropathy in a glucose-independent manner. Here we compared the effects of the DPP-4 inhibitor linagliptin versus telmisartan in preventing CKD progression in non-diabetic rats with 5/6 nephrectomy. Animals were allocated to 1 of 4 groups: sham operated plus placebo; 5/6 nephrectomy plus placebo; 5/6 nephrectomy plus linagliptin; and 5/6 nephrectomy plus telmisartan. Interstitial fibrosis was significantly decreased by 48% with linagliptin but a non-significant 24% with telmisartan versus placebo. The urine albumin-to-creatinine ratio was significantly decreased by 66% with linagliptin and 92% with telmisartan versus placebo. Blood pressure was significantly lowered by telmisartan, but it was not affected by linagliptin. As shown by mass spectrometry, the number of altered peptide signals for linagliptin in plasma was 552 and 320 in the kidney. For telmisartan, there were 108 peptide changes in plasma and 363 in the kidney versus placebo. Linagliptin up-regulated peptides derived from collagen type I, apolipoprotein C1, and heterogeneous nuclear ribonucleoproteins A2/B1, a potential downstream target of atrial natriuretic peptide, whereas telmisartan up-regulated angiotensin II. A second study was conducted to confirm these findings in 5/6 nephrectomy wild-type and genetically deficient DPP-4 rats treated with linagliptin or placebo. Linagliptin therapy in wild-type rats was as effective as DPP-4 genetic deficiency in terms of albuminuria reduction. Thus, linagliptin showed comparable efficacy to telmisartan in preventing CKD progression in non-diabetic rats with 5/6 nephrectomy. However, the underlying pathways seem to be different. Copyright (C) 2016, International Society of Nephrology. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). KW - albuminuria KW - angiotensin receptor blockers KW - chronic kidney disease KW - DPP-4 inhibition KW - proteinuria KW - proteomic analysis Y1 - 2016 U6 - https://doi.org/10.1016/j.kint.2016.01.016 SN - 0085-2538 SN - 1523-1755 VL - 89 SP - 1049 EP - 1061 PB - Nature Publ. Group CY - New York ER - TY - JOUR A1 - Chaykovska, Lyubov A1 - Heunisch, Fabian A1 - von Einem, Gina A1 - Alter, Markus L. A1 - Hocher, Carl-Friedrich A1 - Tsuprykov, Oleg A1 - Dschietzig, Thomas A1 - Kretschmer, Axel A1 - Hocher, Berthold T1 - Urinary Vitamin D Binding Protein and KIM-1 Are Potent New Biomarkers of Major Adverse Renal Events in Patients Undergoing Coronary Angiography JF - PLoS one N2 - Background Vitamin-D-binding protein (VDBP) is a low molecular weight protein that is filtered through the glomerulus as a 25-(OH) vitamin D 3/VDBP complex. In the normal kidney VDBP is reabsorbed and catabolized by proximal tubule epithelial cells reducing the urinary excretion to trace amounts. Acute tubular injury is expected to result in urinary VDBP loss. The purpose of our study was to explore the potential role of urinary VDBP as a biomarker of an acute renal damage. Method We included 314 patients with diabetes mellitus or mild renal impairment undergoing coronary angiography and collected blood and urine before and 24 hours after the CM application. Patients were followed for 90 days for the composite endpoint major adverse renal events (MARE: need for dialysis, doubling of serum creatinine after 90 days, unplanned emergency rehospitalization or death). Results Increased urine VDBP concentration 24 hours after contrast media exposure was predictive for dialysis need (no dialysis: 113.06 +/- 299.61ng/ml, n = 303; need for dialysis: 613.07 +/- 700.45 ng/ml, n = 11, Mean +/- SD, p < 0.001), death (no death during follow-up: 121.41 +/- 324.45 ng/ml, n = 306; death during follow-up: 522.01 +/- 521.86 ng/ml, n = 8; Mean +/- SD, p < 0.003) and MARE (no MARE: 112.08 +/- 302.00ng/ml, n = 298; MARE: 506.16 +/- 624.61 ng/ml, n = 16, Mean +/- SD, p < 0.001) during the follow-up of 90 days after contrast media exposure. Correction of urine VDBP concentrations for creatinine excretion confirmed its predictive value and was consistent with increased levels of urinary Kidney Injury Molecule1 (KIM-1) and baseline plasma creatinine in patients with above mentioned complications. The impact of urinary VDBP and KIM-1 on MARE was independent of known CIN risk factors such as anemia, preexisting renal failure, preexisting heart failure, and diabetes. Conclusions Urinary VDBP is a promising novel biomarker of major contrast induced nephropathy-associated events 90 days after contrast media exposure. Y1 - 2016 U6 - https://doi.org/10.1371/journal.pone.0145723 SN - 1932-6203 VL - 11 PB - PLoS CY - San Fransisco ER - TY - JOUR A1 - Swidsinski, Alexander A1 - Loening-Baucke, Vera A1 - Schulz, Stefan A1 - Manowsky, Julia A1 - Verstraelen, Hans A1 - Swidsinski, Sonja T1 - Functional anatomy of the colonic bioreactor: Impact of antibiotics and Saccharomyces boulardii on bacterial composition in human fecal cylinders JF - Systematic and Applied Microbiology N2 - Sections of fecal cylinders were analyzed using fluorescence in situ hybridization targeting 180 bacterial groups. Samples were collected from three groups of women (N = 20 each) treated for bacterial vaginosis with ciprofloxacin + metronidazole. Group A only received the combined antibiotic regimen, whereas the A/Sb group received concomitant Saccharomyces boulardii CNCM I-745 treatment, and the A.Sb group received S. boulardii prophylaxis following the 14-day antibiotic course. The number of stool cylinders analyzed was 188 out of 228 in group A, 170 out of 228 in group A/Sb, and 172 out of 216 in group Ash. The colonic biomass was organized into a separate mucus layer with no bacteria, a 10-30 mu m broad unstirred transitional layer enriched with bacteria, and a patchy fermentative area that mixed digestive leftovers with bacteria. The antibiotics suppressed bacteria mainly in the fermentative area, whereas abundant bacterial clades retreated to the transitional mucus and survived. As a result, the total concentration of bacteria decreased only by one order. These effects were lasting, since the overall recovery of the microbial mass, bacterial diversity and concentrations were still below pre-antibiotic values 4 months after the end of antibiotic treatment. Sb-prophylaxis markedly reduced antibiotic effects and improved the recovery rates. Since the colon is a sophisticated bioreactor, the study indicated that the spatial anatomy of its biomass was crucial for its function. (C) 2015 The Authors. Published by Elsevier GmbH. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). KW - FISH KW - Colonic microbiota KW - Antibiotics KW - Bioreactor KW - Saccharomyces boulardii KW - Clostridium difficile Y1 - 2016 U6 - https://doi.org/10.1016/j.syapm.2015.11.002 SN - 0723-2020 VL - 39 SP - 67 EP - 75 PB - Nature Publ. Group CY - Jena ER - TY - JOUR A1 - Lu, Yong Ping A1 - Tsuprykov, Oleg A1 - Vignon-Zellweger, Nicolas A1 - Heiden, Susi A1 - Hocher, Berthold T1 - Global Overexpression of ET-1 Decreases Blood Pressure - A Systematic Review and Meta-Analysis of ET-1 Transgenic Mice JF - Naunyn-Schmiedeberg's archives of pharmacology N2 - Background/Aims: ET-1 has independent effects on blood pressure regulation in vivo, it is involved in tubular water and salt excretion, promotes constriction of smooth muscle cells, modulates sympathetic nerve activity, and activates the liberation of nitric oxide. To determine the net effect of these partially counteracting mechanisms on blood pressure, a systematic meta-analysis was performed. Methods: Based on the principles of Cochrane systematic reviews, we searched in major literature databases - MEDLINE (PubMed), Embase, Google Scholar, and the China Biological Medicine Database (CBM-disc) - for articles relevant to the topic of the blood pressure phenotype of endothelin-1 transgenic (ET-1+/+) mice from January 1, 1988 to March 31, 2016. Review Manager Version 5.0 (Rev-Man 5.0) software was applied for statistical analysis. In total thirteen studies reported blood pressure data. Results: The meta-analysis of blood pressure data showed that homozygous ET-1 transgenic mice (ET-1+/+ mice) had a significantly lower blood pressure as compared to WT mice (mean difference:-2.57 mmHg, 95% CI: -4.98 similar to -0.16, P = 0.04), with minimal heterogeneity (P = 0.86). A subgroup analysis of mice older than 6 months revealed that the blood pressure difference between ET-1+/+ mice and WT mice was even more pronounced (mean difference: -6.19 mmHg, 95% CI: -10.76 similar to -1.62, P = 0.008), with minimal heterogeneity (P = 0.91). Conclusion: This meta-analysis provides robust evidence that global ET-1 overexpression in mice lowers blood pressure in an age-dependent manner. Older ET-1+/+ mice have a somewhat more pronounced reduction of blood pressure. (C) 2016 The Author(s) Published by S. Karger AG, Basel KW - Endothelin-1 transgenic mice KW - Blood pressure KW - Kidney weight KW - Heart weight Y1 - 2016 U6 - https://doi.org/10.1159/000450567 SN - 1420-4096 SN - 1423-0143 VL - 41 SP - 770 EP - 780 PB - Karger CY - Basel ER - TY - JOUR A1 - Uhr, Linda A1 - Wieland, Phillis A1 - Homann, Thomas A1 - Huschek, Gerd A1 - Rawel, Harshadrai Manilal T1 - Identification and LC-MS/MS-based analyses of technical enzymes in wheat flour and baked products JF - European food research and technology : official organ of the EuCheMS, Division of Food Chemistry N2 - The use of technical enzymes in bakery industry is necessary for a consistent and good quality of baked products. Since the cultivation of cereals leads to low amounts of endogenous enzymes being present, a need of their commercial alternatives is becoming a routine process in order to meet the consumer quality demands. Targeted quantification proteomics-based methods are necessary for their detection to meet the regulatory criteria. Here, we initially report on the identification of Lipase FE-01, a lipase from fungus Thermomyces lanuginosus, as analyzed by SDS-PAGE, in-Gel digestion, and MALDI-TOF-MS. In further experiments, the focus of the study was directed toward an extensive use and optimization of in-solution enzymatic digestion in combination with LC-MS/MS techniques in identification of specific peptide markers and finally in utilization of the latter in delivering reproducible quantification data for several different technical enzymes (alpha-amylases, xylanase, and lipases from microbial origin) in complex matrices such as baked bread and wheat flour. Two digestion protocols (a fast option using thermocycler program and the well-established overnight method) were tested, and both of these can be successfully applied. The application of isotopically labeled analogs of the MRM targeted peptides as internal standards and the addition of an internal protein standard during the extraction/digestion experiment were compared to determine the optimal quantification algorithm of the recovered enzyme concentrations. Thus, a standardized sensitive LC-MS/MS method could be developed to determine technical enzymes as forthcoming ingredients in the prefabricated food formulations in concentrations lower than 10 ppm. KW - Technical enzymes KW - Amylase KW - Xylanase KW - Lipase KW - Baked products KW - Mass spectrometry Y1 - 2016 U6 - https://doi.org/10.1007/s00217-015-2536-5 SN - 1438-2377 SN - 1438-2385 VL - 242 SP - 247 EP - 257 PB - Springer CY - New York ER - TY - THES A1 - Weber, Florian Till T1 - In vitro Toxizität von Elektrolytlösungen der Lithium-Ionen Batterie Y1 - 2016 ER - TY - THES A1 - Markova, Mariya T1 - Metabolic and molecular effects of two different isocaloric high protein diets in subjects with type 2 diabetes T1 - Metabolische und molekulare Effekte zweier isokalorischer Hochproteindiäten unterschiedlicher Herkunft bei Probanden mit Typ-2-Diabetes N2 - Ernährung stellt ein wichtiger Faktor in der Prävention und Therapie von Typ-2-Diabetes dar. Frühere Studien haben gezeigt, dass Hochproteindiäten sowohl positive als auch negative Effekte auf den Metabolismus hervorrufen. Jedoch ist unklar, ob die Herkunft des Proteins dabei eine Rolle spielt. In der LeguAN-Studie wurden die Effekte von zwei unterschiedlichen Hochproteindiäten, entweder tierischer oder pflanzlicher Herkunft, bei Typ-2-Diabetes Patienten untersucht. Beide Diäten enthielten 30 EN% Proteine, 40 EN% Kohlenhydrate und 30 EN% Fette. Der Anteil an Ballaststoffen, der glykämischer Index und die Fettkomposition waren in beiden Diäten ähnlich. Die Proteinaufnahme war höher, während die Fettaufnahme niedriger im Vergleich zu den früheren Ernährungsgewohnheiten der Probanden war. Insgesamt führten beide Diätinterventionen zu einer Verbesserung der glykämischen Kontrolle, der Insulinsensitivität, des Leberfettgehalts und kardiovaskulärer Risikomarkern ohne wesentliche Unterschiede zwischen den Proteintypen. In beiden Interventionsgruppen wurden die nüchternen Glukosewerte zusammen mit Indizes von Insulinresistenz in einem unterschiedlichen Ausmaß, jedoch ohne signifikante Unterschiede zwischen beiden Diäten verbessert. Die Reduktion von HbA1c war ausgeprägter in der pflanzlichen Gruppe, während sich die Insulinsensitivität mehr in der tierischen Gruppe erhöhte. Die Hochproteindiäten hatten nur einen geringfügigen Einfluss auf den postprandialen Metabolismus. Dies zeigte sich durch eine leichte Verbesserung der Indizes für Insulinsekretion, -sensitivität und –degradation sowie der Werte der freien Fettsäuren. Mit Ausnahme des Einflusses auf die GIP-Sekretion riefen die tierische und die pflanzliche Testmahlzeit ähnliche metabolische und hormonelle Antworten, trotz unterschiedlicher Aminosäurenzusammensetzung. Die tierische Hochproteindiät führte zu einer selektiven Zunahme der fettfreien Masse und Abnahme der Fettmasse, was nicht signifikant unterschiedlich von der pflanzlichen Gruppe war. Darüber hinaus reduzierten die Hochproteindiäten den Leberfettgehalt um durchschnittlich 42%. Die Reduktion des Leberfettgehaltes ging mit einer Verminderung der Lipogenese, der Lipolyse und des freien Fettsäure Flux einher. Beide Interventionen induzierten einen moderaten Abfall von Leberenzymen im Blut. Die Reduktion des Leberfetts war mit einer verbesserten Glukosehomöostase und Insulinsensitivität assoziiert. Blutlipide sanken in allen Probanden, was eventuell auf die niedrigere Fettaufnahme zurückzuführen war. Weiterhin waren die Spiegel an Harnsäure und Inflammationsmarkern erniedrigt unabhängig von der Proteinquelle. Die Werte des systolischen und diastolischen Blutdrucks sanken nur in der pflanzlichen Gruppe, was auf eine potentielle Rolle von Arginin hinweist. Es wurden keine Hinweise auf eine beeinträchtigte Nierenfunktion durch die 6-wöchige Hochproteindiäten beobachtet unabhängig von der Herkunft der Proteine. Serumkreatinin war nur in der pflanzlichen Gruppe signifikant reduziert, was eventuell an dem geringen Kreatingehalt der pflanzlichen Nahrungsmittel liegen könnte. Jedoch sind längere Studien nötig, um die Sicherheit von Hochproteindiäten vollkommen aufklären zu können. Des Weiteren verursachte keine der Diäten eine Induktion des mTOR Signalwegs weder im Fettgewebe noch in Blutzellen. Die Verbesserung der Ganzkörper-Insulinsensitivität deutete auch auf keine Aktivierung von mTOR und keine Verschlechterung der Insulinsensitivität im Skeletmuskel hin. Ein nennenswerter Befund war die erhebliche Reduktion von FGF21, einem wichtigen Regulator metabolischer Prozesse, um ungefähr 50% bei beiden Proteinarten. Ob hepatischer ER-Stress, Ammoniumniveau oder die Makronährstoffpräferenz hinter dem paradoxen Ergebnis stehen, sollte weiter im Detail untersucht werden. Entgegen der anfänglichen Erwartung und der bisherigen Studienlage zeigte die pflanzlich-betonte Hochproteindiät keine klaren Vorteile gegenüber der tierischen Diät. Der ausgeprägte günstige Effekt des tierischen Proteins auf Insulinhomöostase trotz des hohen BCAA-Gehaltes war sicherlich unerwartet und deutet darauf hin, dass bei dem längeren Verzehr andere komplexe metabolische Adaptationen stattfinden. Einen weiteren Aspekt stellt der niedrigere Fettverzehr dar, der eventuell auch zu den Verbesserungen in beiden Gruppen beigetragen hat. Zusammenfassend lässt sich sagen, dass eine 6-wöchige Diät mit 30 EN% Proteinen (entweder pflanzlich oder tierisch), 40 EN% Kohlenhydraten und 30 EN% Fetten mit weniger gesättigten Fetten zu metabolischen Verbesserungen bei Typ-2-Diabetes Patienten unabhängig von Proteinherkunft führt. N2 - Dietary approaches contribute to the prevention and treatment of type 2 diabetes. High protein diets were shown to exert beneficial as well as adverse effects on metabolism. However, it is unclear whether the protein origin plays a role in these effects. The LeguAN study investigated in detail the effects of two high protein diets, either from plant or animal origin, in type 2 diabetic patients. Both diets contained 30 EN% protein, 40 EN% carbohydrates, and 30 EN% fat. Fiber content, glycemic index, and composition of dietary fats were similar in both diets. In comparison to previous dietary habits, the fat content was exchanged for protein, while the carbohydrate intake was not modified. Overall, both high protein diets led to improvements of glycemic control, insulin sensitivity, liver fat, and cardiovascular risk markers without remarkable differences between the protein types. Fasting glucose together with indices of insulin resistance were ameliorated by both interventions to varying extents but without significant differences between protein types. The decline of HbA1c was more pronounced in the plant protein group, whereby the improvement of insulin sensitivity in the animal protein group. The high protein intake had only slight influence on postprandial metabolism seen for free fatty acids and indices of insulin secretion, sensitivity and degradation. Except for GIP release, ingestion of animal and plant meals did not provoke differential metabolic and hormonal responses despite diverse circulating amino acid levels. The animal protein diets led to a selective increase of fat-free mass and decrease of total fat mass, which was not significantly different from the plant protein diet. Moreover, the high protein diets potently decreased liver fat content by 42% on average which was linked to significantly diminished lipogenesis, free fatty acids flux and lipolysis in adipose tissue. Moderate decline of circulating liver enzymes was induced by both interventions. The liver fat reduction was associated with improved glucose homeostasis and insulin sensitivity which underlines the protective effect of the diets. Blood lipid profile improved in all subjects and was probably related to the lower fat intake. Reductions in uric acid and markers of inflammation further argued for metabolic benefits of both high protein diets. Systolic and diastolic blood pressure declined only in the PP group pointing a possible role of arginine. Kidney function was not altered by high protein consumption over 6 weeks. The rapid decrease of serum creatinine in the PP group was noteworthy and should be further investigated. Protein type did not seem to play a role but long-term studies are warranted to fully elucidate safety of high protein regimen. Varying the source of dietary proteins did not affect the mTOR pathway in adipose tissue and blood cells under neither acute nor chronic settings. Enhancement of whole-body insulin sensitivity suggested also no alteration of mTOR and no impairment of insulin sensitivity in skeletal muscle. A remarkable outcome was the extensive reduction of FGF21, critical regulator of metabolic processes, by approximately 50% independently of protein type. Whether hepatic ER-stress, ammonia flux or rather macronutrient preferences is behind this paradoxical finding remains to be investigated in detail. Unlike initial expectations and previous reports plant protein based diet had no clear advantage over animal proteins. The pronounced beneficial effect of animal protein on insulin homeostasis despite high BCAA and methionine intake was certainly unexpected assuming more complex metabolic adaptations occurring upon prolonged consumption. In addition, the reduced fat intake may have also contributed to the overall improvements in both groups. Taking into account the above observed study results, a short-term diet containing 30 EN% protein (either from plant or animal origin), 40 EN% carbohydrates, and 30 EN% fat with lower SFA amount leads to metabolic improvements in diabetic patients, regardless of protein source. KW - type 2 diabetes KW - nutrition KW - protein KW - Typ-2-Diabetes KW - Ernährung KW - high protein diet Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-394310 ER - TY - THES A1 - Ambrosi, Thomas H. T1 - The Role of Bone-residing Adipocyte Progenitors in Age-related Stem Cell Dysfunction and Regenerative Processes Y1 - 2016 ER - TY - THES A1 - Hallahan, Nicole T1 - Identification and characterization of a T2D QTL arising from an NZO.DBA mouse cross Y1 - 2016 ER - TY - JOUR A1 - Bzymek, Robert A1 - Horsthemke, Markus A1 - Isfort, Katrin A1 - Mohr, Simon A1 - Tjaden, Kerstin A1 - Mueller-Tidow, Carsten A1 - Thomann, Marlies A1 - Schwerdtle, Tanja A1 - Baehler, Martin A1 - Schwab, Albrecht A1 - Hanley, Peter J. T1 - Real-time two- and three-dimensional imaging of monocyte motility and navigation on planar surfaces and in collagen matrices: roles of Rho JF - Scientific reports N2 - We recently found that macrophages from RhoA/RhoB double knockout mice had increased motility of the cell body, but severely impaired retraction of the tail and membrane extensions, whereas RhoA-or RhoB-deficient cells exhibited mild phenotypes. Here we extended this work and investigated the roles of Rho signaling in primary human blood monocytes migrating in chemotactic gradients and in various settings. Monocyte velocity, but not chemotactic navigation, was modestly dependent on Rho-ROCK-myosin II signaling on a 2D substrate or in a loose collagen type I matrix. Viewed by time-lapse epi-fluorescence microscopy, monocytes appeared to flutter rather than crawl, such that the 3D surface topology of individual cells was difficult to predict. Spinning disk confocal microscopy and 3D reconstruction revealed that cells move on planar surfaces and in a loose collagen matrix using prominent, curved planar protrusions, which are rapidly remodeled and reoriented, as well as resorbed. In a dense collagen type I matrix, there is insufficient space for this mode and cells adopt a highly Rho-dependent, lobular mode of motility. Thus, in addition to its role in tail retraction on 2D surfaces, Rho is critical for movement in confined spaces, but is largely redundant for motility and chemotaxis in loose matrices. Y1 - 2016 U6 - https://doi.org/10.1038/srep25016 SN - 2045-2322 VL - 6 PB - Nature Publ. Group CY - London ER - TY - GEN A1 - Walter, Tim A1 - Collenburg, Lena A1 - Japtok, Lukasz A1 - Kleuser, Burkhard A1 - Schneider-Schaulies, Sibylle A1 - Müller, Nora A1 - Becam, Jerome A1 - Schubert-Unkmeir, Alexandra A1 - Kong, Ji Na A1 - Bieberich, Erhard A1 - Seibel, Jürgen T1 - Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells N2 - The synthesis and biological evaluation of azido-N-oleoyl serinol is reported. It mimicks biofunctional lipid ceramides and has shown to be capable of click reactions for cell membrane imaging in Jurkat and human brain microvascular endothelial cells. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 324 Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-394960 ER - TY - JOUR A1 - Collenburg, Lena A1 - Walter, Tim A1 - Burgert, Anne A1 - Mueller, Nora A1 - Seibel, Juergen A1 - Japtok, Lukasz A1 - Kleuser, Burkhard A1 - Sauer, Markus A1 - Schneider-Schaulies, Sibylle T1 - A Functionalized Sphingolipid Analogue for Studying Redistribution during Activation in Living T Cells JF - The journal of immunology N2 - Sphingolipids are major components of the plasma membrane. In particular, ceramide serves as an essential building hub for complex sphingolipids, but also as an organizer of membrane domains segregating receptors and signalosomes. Sphingomyelin breakdown as a result of sphingomyelinase activation after ligation of a variety of receptors is the predominant source of ceramides released at the plasma membrane. This especially applies to T lymphocytes where formation of ceramide-enriched membrane microdomains modulates TCR signaling. Because ceramide release and redistribution occur very rapidly in response to receptor ligation, novel tools to further study these processes in living T cells are urgently needed. To meet this demand, we synthesized nontoxic, azido-functionalized ceramides allowing for bio-orthogonal click-reactions to fluorescently label incorporated ceramides, and thus investigate formation of ceramide-enriched domains. Azido-functionalized C-6-ceramides were incorporated into and localized within plasma membrane microdomains and proximal vesicles in T cells. They segregated into clusters after TCR, and especially CD28 ligation, indicating efficient sorting into plasma membrane domains associated with T cell activation; this was abolished upon sphingomyelinase inhibition. Importantly, T cell activation was not abrogated upon incorporation of the compound, which was efficiently excluded from the immune synapse center as has previously been seen in Ab-based studies using fixed cells. Therefore, the functionalized ceramides are novel, highly potent tools to study the subcellular redistribution of ceramides in the course of T cell activation. Moreover, they will certainly also be generally applicable to studies addressing rapid stimulation-mediated ceramide release in living cells. Y1 - 2016 U6 - https://doi.org/10.4049/jimmunol.1502447 SN - 0022-1767 SN - 1550-6606 VL - 196 SP - 3951 EP - 3962 PB - American Assoc. of Immunologists CY - Bethesda ER - TY - JOUR A1 - Walter, T. A1 - Collenburg, Lena A1 - Japtok, Lukasz A1 - Kleuser, Burkhard A1 - Schneider-Schaulies, Sibylle A1 - Mueller, N. A1 - Becam, Jerome A1 - Schubert-Unkmeir, A. A1 - Kong, J. N. A1 - Bieberich, Erhard A1 - Seibel, J. T1 - Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells JF - Chemical communications N2 - The synthesis and biological evaluation of azido-N-oleoyl serinol is reported. It mimicks biofunctional lipid ceramides and has shown to be capable of click reactions for cell membrane imaging in Jurkat and human brain microvascular endothelial cells. Y1 - 2016 U6 - https://doi.org/10.1039/c6cc02879a SN - 1359-7345 SN - 1364-548X VL - 52 SP - 8612 EP - 8614 PB - Royal Society of Chemistry CY - Cambridge ER - TY - THES A1 - Errard, Audrey T1 - Multiple-pest infestations: impact on tomato Solanum lycopersicum biochemistry in the presence/absence of predator, and on pest biology Y1 - 2016 ER - TY - GEN A1 - Errard, Audrey A1 - Ulrichs, Christian A1 - Kühne, Stefan A1 - Mewis, Inga A1 - Mishig, Narantuya A1 - Maul, Ronald A1 - Drungowski, Mario A1 - Parolin, Pia A1 - Schreiner, Monika A1 - Baldermann, Susanne T1 - Metabolite profiling reveals a specific response in tomato to predaceous Chrysoperla carnea larvae and herbivore(s)-predator interactions with the generalist pests Tetranychus urticae and Myzus persicae T2 - Frontiers in plant science N2 - The spider mite Tetranychus urticae Koch and the aphid Myzus persicae (Sulzer) both infest a number of economically significant crops, including tomato (Solanurn lycopersicum). Although used for decades to control pests, the impact of green lacewing larvae Chrysoperla carnea (Stephens) on plant biochemistry was not investigated. Here, we used profiling methods and targeted analyses to explore the impact of the predator and herbivore(s)-predator interactions on tomato biochemistry. Each pest and pest -predator combination induced a characteristic metabolite signature in the leaf and the fruit thus, the plant exhibited a systemic response. The treatments had a stronger impact on non-volatile metabolites including abscisic acid and amino acids in the leaves in comparison with the fruits. In contrast, the various biotic factors had a greater impact on the carotenoids in the fruits. We identified volatiles such as myrcene and alpha-terpinene which were induced by pest -predator interactions but not by single species, and we demonstrated the involvement of the phytohormone abscisic acid in tritrophic interactions for the first time. More importantly, C. carnea larvae alone impacted the plant metabolome, but the predator did not appear to elicit particular defense pathways on its own. Since the presence of both C. carnea larvae and pest individuals elicited volatiles which were shown to contribute to plant defense, C. carnea larvae could therefore contribute to the reduction of pest infestation, not only by its preying activity, but also by priming responses to generalist herbivores such as T urticae and M. persicae. On the other hand, the use of C. carnea larvae alone did not impact carotenoids thus, was not prejudicial to the fruit quality. The present piece of research highlights the specific impact of predator and tritrophic interactions with green lacewing larvae, spider mites, and aphids on different components of the tomato primary and secondary metabolism for the first time, and provides cues for further in-depth studies aiming to integrate entomological approaches and plant biochemistry. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 454 KW - carotenoids KW - plant volatiles KW - Chrysopidae KW - Solanaceae KW - multiple-pest infestation KW - tritrophic system KW - Twister TM KW - biological pest control Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-407913 ER -