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Anthropogenic climate change alters the hydrological cycle. While certain areas experience more intense precipitation events, others will experience droughts and increased evaporation, affecting water storage in long-term reservoirs, groundwater, snow, and glaciers. High elevation environments are especially vulnerable to climate change, which will impact the water supply for people living downstream. The Himalaya has been identified as a particularly vulnerable system, with nearly one billion people depending on the runoff in this system as their main water resource. As such, a more refined understanding of spatial and temporal changes in the water cycle in high altitude systems is essential to assess variations in water budgets under different climate change scenarios.
However, not only anthropogenic influences have an impact on the hydrological cycle, but changes to the hydrological cycle can occur over geological timescales, which are connected to the interplay between orogenic uplift and climate change. However, their temporal evolution and causes are often difficult to constrain. Using proxies that reflect hydrological changes with an increase in elevation, we can unravel the history of orogenic uplift in mountain ranges and its effect on the climate.
In this thesis, stable isotope ratios (expressed as δ2H and δ18O values) of meteoric waters and organic material are combined as tracers of atmospheric and hydrologic processes with remote sensing products to better understand water sources in the Himalayas. In addition, the record of modern climatological conditions based on the compound specific stable isotopes of leaf waxes (δ2Hwax) and brGDGTs (branched Glycerol dialkyl glycerol tetraethers) in modern soils in four Himalayan river catchments was assessed as proxies of the paleoclimate and (paleo-) elevation. Ultimately, hydrological variations over geological timescales were examined using δ13C and δ18O values of soil carbonates and bulk organic matter originating from sedimentological sections from the pre-Siwalik and Siwalik groups to track the response of vegetation and monsoon intensity and seasonality on a timescale of 20 Myr.
I find that Rayleigh distillation, with an ISM moisture source, mainly controls the isotopic composition of surface waters in the studied Himalayan catchments. An increase in d-excess in the spring, verified by remote sensing data products, shows the significant impact of runoff from snow-covered and glaciated areas on the surface water isotopic values in the timeseries.
In addition, I show that biomarker records such as brGDGTs and δ2Hwax have the potential to record (paleo-) elevation by yielding a significant correlation with the temperature and surface water δ2H values, respectively, as well as with elevation. Comparing the elevation inferred from both brGDGT and δ2Hwax, large differences were found in arid sections of the elevation transects due to an additional effect of evapotranspiration on δ2Hwax. A combined study of these proxies can improve paleoelevation estimates and provide recommendations based on the results found in this study.
Ultimately, I infer that the expansion of C4 vegetation between 20 and 1 Myr was not solely dependent on atmospheric pCO2, but also on regional changes in aridity and seasonality from to the stable isotopic signature of the two sedimentary sections in the Himalaya (east and west).
This thesis shows that the stable isotope chemistry of surface waters can be applied as a tool to monitor the changing Himalayan water budget under projected increasing temperatures. Minimizing the uncertainties associated with the paleo-elevation reconstructions were assessed by the combination of organic proxies (δ2Hwax and brGDGTs) in Himalayan soil. Stable isotope ratios in bulk soil and soil carbonates showed the evolution of vegetation influenced by the monsoon during the late Miocene, proving that these proxies can be used to record monsoon intensity, seasonality, and the response of vegetation. In conclusion, the use of organic proxies and stable isotope chemistry in the Himalayas has proven to successfully record changes in climate with increasing elevation. The combination of δ2Hwax and brGDGTs as a new proxy provides a more refined understanding of (paleo-)elevation and the influence of climate.
Zinc is an essential trace element, making it crucial to have a reliable biomarker for evaluating an individual’s zinc status. The total serum zinc concentration, which is presently the most commonly used biomarker, is not ideal for this purpose, but a superior alternative is still missing. The free zinc concentration, which describes the fraction of zinc that is only loosely bound and easily exchangeable, has been proposed for this purpose, as it reflects the highly bioavailable part of serum zinc. This report presents a fluorescence-based method for determining the free zinc concentration in human serum samples, using the fluorescent probe Zinpyr-1. The assay has been applied on 154 commercially obtained human serum samples. Measured free zinc concentrations ranged from 0.09 to 0.42 nM with a mean of 0.22 ± 0.05 nM. It did not correlate with age or the total serum concentrations of zinc, manganese, iron or selenium. A negative correlation between the concentration of free zinc and total copper has been seen for sera from females. In addition, the free zinc concentration in sera from females (0.21 ± 0.05 nM) was significantly lower than in males (0.23 ± 0.06 nM). The assay uses a sample volume of less than 10 µL, is rapid and cost-effective and allows us to address questions regarding factors influencing the free serum zinc concentration, its connection with the body’s zinc status, and its suitability as a future biomarker for an individual’s zinc status.
Background
High blood glucose and diabetes are amongst the conditions causing the greatest losses in years of healthy life worldwide. Therefore, numerous studies aim to identify reliable risk markers for development of impaired glucose metabolism and type 2 diabetes. However, the molecular basis of impaired glucose metabolism is so far insufficiently understood. The development of so called 'omics' approaches in the recent years promises to identify molecular markers and to further understand the molecular basis of impaired glucose metabolism and type 2 diabetes. Although univariate statistical approaches are often applied, we demonstrate here that the application of multivariate statistical approaches is highly recommended to fully capture the complexity of data gained using high-throughput methods.
Methods
We took blood plasma samples from 172 subjects who participated in the prospective Metabolic Syndrome Berlin Potsdam follow-up study (MESY-BEPO Follow-up). We analysed these samples using Gas Chromatography coupled with Mass Spectrometry (GC-MS), and measured 286 metabolites. Furthermore, fasting glucose levels were measured using standard methods at baseline, and after an average of six years. We did correlation analysis and built linear regression models as well as Random Forest regression models to identify metabolites that predict the development of fasting glucose in our cohort.
Results
We found a metabolic pattern consisting of nine metabolites that predicted fasting glucose development with an accuracy of 0.47 in tenfold cross-validation using Random Forest regression. We also showed that adding established risk markers did not improve the model accuracy. However, external validation is eventually desirable. Although not all metabolites belonging to the final pattern are identified yet, the pattern directs attention to amino acid metabolism, energy metabolism and redox homeostasis.
Conclusions
We demonstrate that metabolites identified using a high-throughput method (GC-MS) perform well in predicting the development of fasting plasma glucose over several years. Notably, not single, but a complex pattern of metabolites propels the prediction and therefore reflects the complexity of the underlying molecular mechanisms. This result could only be captured by application of multivariate statistical approaches. Therefore, we highly recommend the usage of statistical methods that seize the complexity of the information given by high-throughput methods.
Frailty and sarcopenia share some underlying characteristics like loss of muscle mass, low muscle strength, and low physical performance. Imaging parameters and functional examinations mainly assess frailty and sarcopenia criteria; however, these measures can have limitations in clinical settings. Therefore, finding suitable biomarkers that reflect a catabolic muscle state e.g. an elevated muscle protein turnover as suggested in frailty, are becoming more relevant concerning frailty diagnosis and risk assessment.
3-Methylhistidine (3-MH) and its ratios 3-MH-to-creatinine (3-MH/Crea) and 3 MH-to-estimated glomerular filtration rate (3-MH/eGFR) are under discussion as possible biomarkers for muscle protein turnover and might support the diagnosis of frailty. However, there is some skepticism about the reliability of 3-MH measures since confounders such as meat and fish intake might influence 3-MH plasma concentrations. Therefore, the influence of dietary habits and an intervention with white meat on plasma 3-MH was determined in young and healthy individuals. In another study, the cross-sectional associations of plasma 3-MH, 3-MH/Crea and 3-MH/eGFR with the frailty status (robust, pre-frail and frail) were investigated.
Oxidative stress (OS) is a possible contributor to frailty development, and high OS levels as well as low micronutrient levels are associated with the frailty syndrome. However, data on simultaneous measures of OS biomarkers together with micronutrients are lacking in studies including frail, pre-frail and robust individuals. Therefore, cross-sectional associations of protein carbonyls (PrCarb), 3-nitrotyrosine (3-NT) and several micronutrients with the frailty status were determined.
A validated UPLC-MS/MS (ultra-performance liquid chromatography tandem mass spectrometry) method for the simultaneous quantification of 3-MH and 1-MH (1 methylhistidine, as marker for meat and fish consumption) was presented and used for further analyses. Omnivores showed higher plasma 3-MH and 1-MH concentrations than vegetarians and a white meat intervention resulted in an increase in plasma 3-MH, 3 MH/Crea, 1-MH and 1-MH/Crea in omnivores. Elevated 3-MH and 3-MH/Crea levels declined significantly within 24 hours after this white meat intervention. Thus, 3-MH and 3-MH/Crea might be used as biomarker for muscle protein turnover when subjects did not consume meat 24 hours prior to blood samplings.
Plasma 3-MH, 3-MH/Crea and 3-MH/eGFR were higher in frail individuals than in robust individuals. Additionally, these biomarkers were positively associated with frailty in linear regression models, and higher odds to be frail were found for every increase in 3 MH and 3-MH/eGFR quintile in multivariable logistic regression models adjusted for several confounders. This was the first study using 3-MH/eGFR and it is concluded that plasma 3-MH, 3-MH/Crea and 3-MH/eGFR might be used to identify frail individuals or individuals at higher risk to be frail, and that there might be threshold concentrations or ratios to support these diagnoses.
Higher vitamin D3, lutein/zeaxanthin, γ-tocopherol, α-carotene, β-carotene, lycopene and β-cryptoxanthin concentrations and additionally lower PrCarb concentrations were found in robust compared to frail individuals in multivariate linear models. Frail subjects had higher odds to be in the lowest than in the highest tertile for vitamin D3 α-tocopherol, α-carotene, β-carotene, lycopene, lutein/zeaxanthin, and β cryptoxanthin, and had higher odds to be in the highest than in the lowest tertile for PrCarb than robust individuals in multivariate logistic regression models. Thus, a low micronutrient together with a high PrCarb status is associated with pre-frailty and frailty.
Primary progressive multiple sclerosis (PPMS) shows a highly variable disease progression with poor prognosis and a characteristic accumulation of disabilities in patients. These hallmarks of PPMS make it difficult to diagnose and currently impossible to efficiently treat. This study aimed to identify plasma metabolite profiles that allow diagnosis of PPMS and its differentiation from the relapsing remitting subtype (RRMS), primary neurodegenerative disease (Parkinson’s disease, PD), and healthy controls (HCs) and that significantly change during the disease course and could serve as surrogate markers of multiple sclerosis (MS)-associated neurodegeneration over time. We applied untargeted high-resolution metabolomics to plasma samples to identify PPMS-specific signatures, validated our findings in independent sex- and age-matched PPMS and HC cohorts and built discriminatory models by partial least square discriminant analysis (PLS-DA). This signature was compared to sex- and age-matched RRMS patients, to patients with PD and HC. Finally, we investigated these metabolites in a longitudinal cohort of PPMS patients over a 24-month period. PLS-DA yielded predictive models for classification along with a set of 20 PPMS-specific informative metabolite markers. These metabolites suggest disease-specific alterations in glycerophospholipid and linoleic acid pathways. Notably, the glycerophospholipid LysoPC(20:0) significantly decreased during the observation period. These findings show potential for diagnosis and disease course monitoring, and might serve as biomarkers to assess treatment efficacy in future clinical trials for neuroprotective MS therapies.
In dieser Arbeit steht die Entwicklung einer Sensorplattform für biochemische Anwendungen, welche auf einem optischen Detektionsprinzips beruht, im Vordergrund. Während der Entwicklung wurden zwei komplementäre Konzeptideen behandelt, zum einen ein Sensor, der auf photonischen Kristallen und Wellenleiterstrukturen basiert und zum anderen einen faserbasierten Sensor, der chemisch modifizierte Faser-Bragg-Gitter enthält. Das optische Detektionsprinzip in beiden Sensorideen ist die resultierende Brechungsindexänderung als messbare physikochemische Kenngröße.
Das aus der Natur bekannte Phänomen der photonischen Kristalle, das u. a. bei Opalen und bei Schmetterlingen zu finden ist, wurde bereits 1887 von Lord Rayleigh beschrieben. Er beschrieb die optischen Eigenschaften von periodischen mehrschichtigen Filmen, welche als vereinfachtes Modell eines eindimensionalen photonischen Kristalls verstanden werden können. Die Periodizität der Brechungsindexänderung resultiert in einem optischen Filter für Frequenzen in einem bestimmten spektralen Bereich, weshalb dann dort keine Lichtausbreitung mehr möglich ist. Wird dieses System aber durch eine Defektstelle in der Brechungsindexperiodizität gestört, sodass daraus zwei perfekt periodische Systeme entstehen, ist die Lichtausbreitung für eine bestimmte Frequenz dennoch möglich. In der Folge resultiert daraus ein schmalbandiges Signal im Transmissionsspektrum. Die erlaubte Frequenz ist dabei u. a. abhängig vom Brechungsindexunterschied des periodischen Systems, d.h. Veränderung des Brechungsindexes einer Schicht führt zu einer spektralen Verschiebung der erlaubten Frequenz, dadurch kann dieses Sensorkonzept für biochemische Sensorik ausgenutzt werden [1]. Diese Entwicklung des auf photonischen Kristallen basierenden Sensors war eine Kooperation mit dem Industriepartner „Nanoplus GmbH“. In der Doktorarbeit wurden Simulationen und praktischen Arbeiten zur Designentwicklung des Sensors und die Arbeiten an einem ersten Modellaufbau für die biochemischen Anwendungen durchgeführt.
Für den faserbasierten Sensor wurden Faser-Bragg-Gitter in den Faserkern hineingeschrieben. Hill et al. entdeckten 1978, dass solche Gitterstrukturen genau wie photonische Kristalle als optische Filter fungieren [2]. Die Gitter bestehen dabei aus Änderungen des Brechungsindexes im Faserkern. Im Laufe der nächsten vierzig Jahren wurden verschiedene Einschreibetechniken und Gitterstrukturen entwickelt, weshalb die Eigenschaften der jeweiligen Gitterstrukturen variieren. Eine solche Gitterstruktur sind u. a. die Faser-Bragg-Gitter, deren Gitterperiode, d. h. die Abstände der Brechungsindexmodifikationen, sich im Nanometer- bis Mikrometerbereich befinden. Aufgrund der kleinen Gitterperiode wird eine rückwärtsführende Welle im Kern für eine bestimmte Frequenz bzw. Wellenlänge, der Bragg-Wellenlänge, erzeugt. Im Endeffekt resultiert daraus ein schmalbandiges Signal sowohl im Transmissionsspektrum, als auch im Reflexionsspektrum. Die Resonanzwellenlänge ist dabei proportional zu der Gitterperiode und dem effektiven Brechungsindex, welcher vom Brechungsindex des Kerns und des kernumgebenen Materials abhängig ist. Letztlich eignet sich diese Technik für physikochemische Sensorik. Im Rahmen dieser Arbeit wurden die Gitter mit Hilfe einer relativen neuen Herstellungsmethode in die Fasern geschrieben [3]. Anschließend stand die Entwicklung eines Biosensors im Vordergrund, wobei zunächst ein Protokoll zum Ätzen der Faser mit Flusssäure entwickelt worden ist, dass das System sensitiv zum umgebenen Brechungsindex macht. Am Ende wurde ein Modellaufbau realisiert, indem ein Modellsystem, hier die Detektion vom C-reaktiven Protein mittels spezifischen einzelsträngigen DNS-Aptameren, erfolgreich getestet und quantifiziert worden ist.
1 Mandal, S.; Erickson, D. Nanoscale Optofluidic Sensor Arrays. Opt. Express 2008, 16 (3), 1623–1631.
2 Hill, K. O.; Fujii, Y.; Johnson, D. C.; Kawasaki, B. S. Photosensitivity in Optical Fiber Waveguides: Application to Reflection Filter Fabrication. Appl. Phys. Lett. 1978, 32 (10), 647–649.
3 Martínez, A.; Dubov, M.; Khrushchev, I.; Bennion, I. Direct Writing of Fibre Bragg Gratings by Femtosecond Laser. Electron. Lett. 2004, 40 (19), 1170.
Global climate change is one of the greatest challenges of the 21st century, with influence on the environment, societies, politics and economies. The (semi-)arid areas of Southern Africa already suffer from water scarcity. There is a great variety of ongoing research related to global climate history but important questions on regional differences still exist.
In southern African regions terrestrial climate archives are rare, which makes paleoclimate studies challenging. Based on the assumption that continental pans (sabkhas) represent a suitable geo-archive for the climate history, two different pans were studied in the southern and western Kalahari Desert. A combined approach of molecular biological and biogeochemical analyses is utilized to investigate the diversity and abundance of microorganisms and to trace temporal and spatial changes in paleoprecipitation in arid environments. The present PhD thesis demonstrates the applicability of pan sediments as a late Quaternary geo-archive based on microbial signature lipid biomarkers, such as archaeol, branched and isoprenoid glycerol dialkyl glycerol tetraethers (GDGTs) as well as phospholipid fatty acids (PLFA). The microbial signatures contained in the sediment provide information on the current or past microbial community from the Last Glacial Maximum to the recent epoch, the Holocene. The results are discussed in the context of regional climate evolution in southwestern Africa. The seasonal shift of the Innertropical Convergence Zone (ITCZ) along the equator influences the distribution of precipitation- and climate zones. The different expansion of the winter- and summer rainfall zones in southern Africa was confirmed by the frequency of certain microbial biomarkers. A period of increased precipitation in the south-western Kalahari could be described as a result of the extension of the winter rainfall zone during the last glacial maximum (21 ± 2 ka). Instead a period of increased paleoprecipitation in the western Kalahari was indicated during the Late Glacial to Holocene transition. This was possibly caused by a southwestern shift in the position of the summer rainfall zone associated to the southward movement of the ITCZ.
Furthermore, for the first time this study characterizes the bacterial and archaeal life based on 16S rRNA gene high-throughput sequencing in continental pan sediments and provides an insight into the recent microbial community structure. Near-surface processes play an important role for the modern microbial ecosystem in the pans. Water availability as well as salinity might determine the abundance and composition of the microbial communities. The microbial community of pan sediments is dominated by halophilic and dry-adapted archaea and bacteria. Frequently occurring microorganisms such as, Halobacteriaceae, Bacillus and Gemmatimonadetes are described in more detail in this study.
The lakes in the Kenyan Rift Valley offer the unique opportunity to study a wide range of hydrochemical environmental conditions, ranging from freshwater to highly saline and alkaline lakes. Because little is known about the hydro- and biogeochemical conditions in the underlying lake sediments, it was the aim of this study to extend the already existing data sets with data from porewater and biomarker analyses. Additionally, reduced sulphur compounds and sulphate reduction rates in the sediment were determined. The new data was used to examine the anthropogenic and microbial influence on the lakes sediments as well as the influence of the water chemistry on the degradation and preservation of organic matter in the sediment column. The lakes discussed in this study are: Logipi, Eight (a small crater lake in the region of Kangirinyang), Baringo, Bogoria, Naivasha, Oloiden, and Sonachi.
The biomarker compositions were similar in all studied lake sediments; nevertheless, there were some differences between the saline and freshwater lakes. One of those differences is the occurrence of a molecule related to β-carotene, which was only found in the saline lakes. This molecule most likely originates from cyanobacteria, single-celled organisms which are commonly found in saline lakes. In the two freshwater lakes, stigmasterol, a sterol characteristic for freshwater algae, was found. In this study, it was shown that Lakes Bogoria and Sonachi can be used for environmental reconstructions with biomarkers, because the absence of oxygen at the lake bottoms slowed the degradation process. Other lakes, like for example Lake Naivasha, cannot be used for such reconstructions, because of the large anthropogenic influence. But the biomarkers proved to be a useful tool to study those anthropogenic influences. Additionally, it was observed that horizons with a high concentration of elemental sulphur can be used as temporal markers. Those horizons were deposited during times when the lake levels were very low. The sulphur was deposited by microorganisms which are capable of anoxygenic photosynthesis or sulphide oxidation.
Die Qualität von Nutzpflanzen ist von zahlreichen Einflussfaktoren wie beispielsweise Lagerbedingungen und Sorteneigenschaften abhängig. Um Qualitätsmängel zu minimieren und Absatzchancen von Nutzpflanzen zu steigern sind umfangreiche Analysen hinsichtlich ihrer stofflichen Zusammensetzung notwendig. Chromatographische Techniken gekoppelt an ein Massenspektrometer und die Kernspinresonanzspektroskopie wurden dafür bislang verwendet. In der vorliegenden Arbeit wurde ein Gaschromatograph an ein Flugzeitmassenspektrometer (GC-TOF-MS) gekoppelt, um physiologische Prozesse bzw. Eigenschaften (die Schwarzfleckigkeit, die Chipsbräunung, das Physiologische Alter und die Keimhemmung) von Nutzpflanzen aufzuklären. Als Pflanzenmodell wurde dafür die Kartoffelknolle verwendet. Dazu wurden neue analytische Lösungsansätze entwickelt, die eine zielgerichtete Auswertung einer Vielzahl von Proben, die Etablierung einer umfangreichen Referenzspektrenbibliothek und die sichere Archivierung aller experimentellen Daten umfassen. Das Verfahren der Probenvorbereitung wurde soweit modifiziert, dass gering konzentrierte Substanzen mittels GC-TOF-MS analysiert werden können. Dadurch wurde das durch die Probenvorbereitung limitierte Substanzspektrum erweitert. Anhand dieser Lösungsansätze wurden physiologisch relevante Stoffwechselprodukte identifiziert, welche indikativ (klassifizierend) bzw. prädiktiv (vorhersagend) für die physiologischen Prozesse sind. Für die Schwarzfleckigkeitsneigung und die Chipseignung wurde jeweils ein biochemisches Modell zur Vorhersage dieser Prozesse aufgestellt und auf eine Züchtungspopulation übertragen. Ferner wurden für die Schwarzfleckigkeit Stoffwechselprodukte des Respirationsstoffwechsels identifiziert sowie Aminosäuren, Glycerollipide und Phenylpropanoide für das Physiologische Alter als relevant erachtet. Das physiologische Altern konnte durch die Anwendung höherer Temperaturen beschleunigt werden. Durch Anwendung von Keimhemmern (Kümmelöl, Chlorpropham) wurde eine Verzögerung des physiologischen Alterns beobachtet. Die Applikation von Kümmelöl erwies sich dabei als besonders vorteilhaft. Kümmelöl behandelte Knollen wiesen im Vergleich zu unbehandelten Knollen nur Veränderungen im Aminosäure-, Zucker- und Sekundärstoffwechsel auf. Chlorpropham behandelte Knollen wiesen einen ähnlichen Stoffwechsel wie die unbehandelten Knollen auf. Für die bislang noch nicht identifizierten Stoffwechselprodukte wurden im Rahmen dieser Arbeit das Verfahren der „gezielten An-/Abreicherung“, der „gepaarten NMR/GC-TOF-MS Analyse“ und das „Entscheidungsbaumverfahren“ entwickelt. Diese ermöglichen eine Klassifizierung von GC-MS Signalen im Hinblick auf ihre chemische Funktionalität. Das Verfahren der gekoppelten NMR/GC-TOF-MS Analyse erwies sich dabei als besonders erfolgversprechend, da es eine Aufklärung bislang unbekannter gaschromatographischer Signale ermöglicht. In der vorliegenden Arbeit wurden neue Stoffwechselprodukte in der Kartoffelknolle identifiziert, wodurch ein wertvoller Beitrag zur Analytik der Metabolomik geleistet wurde.
The Tibetan Plateau is the largest elevated landmass in the world and profoundly influences atmospheric circulation patterns such as the Asian monsoon system. Therefore this area has been increasingly in focus of palaeoenvironmental studies. This thesis evaluates the applicability of organic biomarkers for palaeolimnological purposes on the Tibetan Plateau with a focus on aquatic macrophyte-derived biomarkers. Submerged aquatic macrophytes have to be considered to significantly influence the sediment organic matter due to their high abundance in many Tibetan lakes. They can show highly 13C-enriched biomass because of their carbon metabolism and it is therefore crucial for the interpretation of δ13C values in sediment cores to understand to which extent aquatic macrophytes contribute to the isotopic signal of the sediments in Tibetan lakes and in which way variations can be explained in a palaeolimnological context. Additionally, the high abundance of macrophytes makes them interesting as potential recorders of lake water δD. Hydrogen isotope analysis of biomarkers is a rapidly evolving field to reconstruct past hydrological conditions and therefore of special relevance on the Tibetan Plateau due to the direct linkage between variations of monsoon intensity and changes in regional precipitation / evaporation balances. A set of surface sediment and aquatic macrophyte samples from the central and eastern Tibetan Plateau was analysed for composition as well as carbon and hydrogen isotopes of n-alkanes. It was shown how variable δ13C values of bulk organic matter and leaf lipids can be in submerged macrophytes even of a single species and how strongly these parameters are affected by them in corresponding sediments. The estimated contribution of the macrophytes by means of a binary isotopic model was calculated to be up to 60% (mean: 40%) to total organic carbon and up to 100% (mean: 66%) to mid-chain n-alkanes. Hydrogen isotopes of n-alkanes turned out to record δD of meteoric water of the summer precipitation. The apparent enrichment factor between water and n-alkanes was in range of previously reported ones (≈-130‰) at the most humid sites, but smaller (average: -86‰) at sites with a negative moisture budget. This indicates an influence of evaporation and evapotranspiration on δD of source water for aquatic and terrestrial plants. The offset between δD of mid- and long-chain n-alkanes was close to zero in most of the samples, suggesting that lake water as well as soil and leaf water are affected to a similar extent by those effects. To apply biomarkers in a palaeolimnological context, the aliphatic biomarker fraction of a sediment core from Lake Koucha (34.0° N; 97.2° E; eastern Tibetan Plateau) was analysed for concentrations, δ13C and δD values of compounds. Before ca. 8 cal ka BP, the lake was dominated by aquatic macrophyte-derived mid-chain n-alkanes, while after 6 cal ka BP high concentrations of a C20 highly branched isoprenoid compound indicate a predominance of phytoplankton. Those two principally different states of the lake were linked by a transition period with high abundances of microbial biomarkers. δ13C values were relatively constant for long-chain n-alkanes, while mid-chain n-alkanes showed variations between -23.5 to -12.6‰. Highest values were observed for the assumed period of maximum macrophyte growth during the late glacial and for the phytoplankton maximum during the middle and late Holocene. Therefore, the enriched values were interpreted to be caused by carbon limitation which in turn was induced by high macrophyte and primary productivity, respectively. Hydrogen isotope signatures of mid-chain n-alkanes have been shown to be able to track a previously deduced episode of reduced moisture availability between ca. 10 and 7 cal ka BP, indicated by a 20‰ shift towards higher δD values. Indications for cooler episodes at 6.0, 3.1 and 1.8 cal ka BP were gained from drops of biomarker concentrations, especially microbial-derived hopanoids, and from coincidental shifts towards lower δ13C values. Those episodes correspond well with cool events reported from other locations on the Tibetan Plateau as well as in the Northern Hemisphere. To conclude, the study of recent sediments and plants improved the understanding of factors affecting the composition and isotopic signatures of aliphatic biomarkers in sediments. Concentrations and isotopic signatures of the biomarkers in Lake Koucha could be interpreted in a palaeolimnological context and contribute to the knowledge about the history of the lake. Aquatic macrophyte-derived mid-chain n-alkanes were especially useful, due to their high abundance in many Tibetan Lakes and their ability to record major changes of lake productivity and palaeo-hydrological conditions. Therefore, they have the potential to contribute to a fuller understanding of past climate variability in this key region for atmospheric circulation systems.
This thesis presents investigations on sediments from two African lakes which have been recording changes in their surrounding environmental and climate conditions since more than 200,000 years. Focus of this work is the time of the last Glacial and the Holocene (the last ~100,000 years before present [in the following 100 kyr BP]). One important precondition for this kind of research is a good understanding of the present ecosystems in and around the lakes and of the sediment formation under modern climate conditions. Both studies therefore include investigations on the modern environment (including organisms, soils, rocks, lake water and sediments). A 90 m long sediment sequence was investigated from Lake Tswaing (north-eastern South Africa) using geochemical analyses. These investigations document alternating periods of high detrital input and low (especially autochthonous) organic matter content and periods of low detrital input, carbonatic or evaporitic sedimentation and high autochthonous organic matter content. These alternations are interpreted as changes between relatively humid and arid conditions, respectively. Before c. 75 kyr BP, they seem to follow changes in local insolation whereas afterwards they appear to be acyclic and are probably caused by changes in ocean circulation and/or in the mean position of the Inter-Tropical Convergence Zone (ITCZ). Today, these factors have main influence on precipitation in this area where rainfall occurs almost exclusively during austral summer. All modern organisms were analysed for their biomarker and bulk organic and compound-specific stable carbon isotope composition. The same investigations on sediments from the modern lake floor document the mixed input of the investigated individual organisms and reveal additional influences by methanotrophic bacteria. A comparison of modern sediment characteristics with those of sediments covering the time 14 to 2 kyr BP shows changes in the productivity of the lake and the surrounding vegetation which are best explained by changes in hydrology. More humid conditions are indicated for times older than 10 kyr BP and younger than 7.5 kyr BP, whereas arid conditions prevailed in between. These observations agree with the results from sediment composition and indications from other climate archives nearby. The second lake study deals with Lake Challa, a small, deep crater lake on the foot of Mount Kilimanjaro. In this lake form mm-scale laminated sediments which were analyses with micro-XRF scanning for changes in the element composition. By comparing these results with investigations on thin sections, results from ongoing sediment trap studies, meteorological data, and investigations on the surrounding rocks and soils, I develop a model for seasonal variability in the limnology and sedimentation of Lake Challa. The lake appears to be stratified during the warm rain seasons (October – December and March – May) during which detrital material is delivered to the lake and carbonates precipitate. On the lake floor forms a dark lamina with high contents of Fe and Ti and high Ca/Al and low Mn/Fe ratios. Diatoms bloom during the cool and windy season (June – September) when mixing down to c. 60 m depth provides easily bio-available nutrients. Contemporaneously, Fe and Mn-oxides are precipitating which cause high Mn/Fe ratios in the light diatom-rich laminae of the sediments. Trends in the Mn/Fe ratio of the sediments are interpreted to reflect changes in the intensity or duration of seasonal mixing in Lake Challa. This interpretation is supported by parallel changes in the organic matter and biogenic silica content observed in the 22 m long profile recovered from Lake Challa. This covers the time of the last 25 kyr BP. It documents a transition around 16 kyr BP from relatively well-mixed conditions with high detrital input during glacial times to stronger stratified conditions which are probably related to increasing lake levels in Challa and generally more humid conditions in East Africa. Intensified mixing is recorded for the time of the Younger Dryas and the period between 11.4 and 10.7 kyr BP. For these periods, reduced intensity of the SW monsoon and intensified NE monsoon are reported from archives of the Indian-Asian Monsoon region, arguing for the latter as a probable source for wind mixing in Lake Challa. This connection is probably also responsible for contemporaneous events in the Mn/Fe ratios of the Lake Challa sediments and in other records of northern hemisphere monsoon intensity during the Holocene and underlines the close interaction of global low latitude atmospheric circulation.
Das seit 1957 als essentiell klassifizierte Spurenelement Selen vermittelt seine Funktion hauptsächlich durch seinen Einbau in Selenoproteine in Form der 21. proteinogenen Aminosäure Selenocystein. Insgesamt wurden 25 humane Gene für Selenoproteine identifiziert, deren genaue Funktion häufig noch nicht bekannt ist. Selen ist das einzige Mitglied aus der Gruppe der Mikronährstoffe, für das nach wie vor eine antikanzerogene Funktion vor allem in Bezug auf Darmkrebs postuliert wird. Die Grundlage dafür liefert eine Interventionsstudie, bei der 1.312 Probanden für 4,5 Jahre mit 200 μg Selen/Tag supplementiert wurden. Dies resultierte in einer Senkung der Gesamtkrebsmortalität um 50 %. Die Fragen einer optimalen Selenzufuhr, die nicht nur den Bedarf deckt, sondern auch die Entfaltung der antikanzerogenen Wirkung von Selen gewährleistet und die zugrunde liegenden molekularen Mechanismen sind noch ungeklärt. Zudem liegt die Selenzufuhr bei einem Großteil der europäischen Bevölkerung unter den Empfehlungen. Deshalb wurden in der vorliegenden Arbeit vier Wochen alte Mäuse für sechs Wochen marginal defizient (0,086 mg/kg Futter) bzw. selenadäquat (0,15 mg/kg Futter) gefüttert. Dieser geringe Unterschied im Selengehalt resultierte in einer Senkung des Plasmaselenspiegels der selenarmen Tiere auf 13 % und der GPx-Aktivität in der Leber auf 35 %. Zunächst wurde der Einfluss von Selen auf die globale Genexpression im murinen Colon mittels Microarray untersucht. Von den im Colon exprimierten Selenoproteinen reagierte die mRNA von SelW, SelH, GPx1 und SelM im Selenmangel besonders deutlich mit Expressionsverlust. Da diese Selenoproteine nicht nur im Colon, sondern auch in Leukozyten reguliert waren, sind sie auch als humane Biomarker für die in dieser Studie gewählte Schwankung des Selengehalts geeignet. Des Weiteren wurde auf Basis der Microarraydaten eine Signalweganalyse durchgeführt, die der Identifizierung krebsrelevanter Signalwege diente, um mögliche molekularbiologische Erklärungsansätze für die Rolle von Selen im Krebsgeschehen zu finden. Es zeigte sich, dass die mRNA von Schlüsselgenen des Wnt-Signalwegs wie β-Catenin, Gsk3β, Dvl2, Tle2, Lef1 und c-Myc auf Schwankungen des Selengehalts reagiert. Vor allem die Induktion von c-Myc, einem Zielgen des Wnt-Signalwegs, deutet darauf hin, dass dieser im Selenmangel tatsächlich aktiver ist als bei selenadäquater Versorgung. Ein weiterer möglicher Erklärungsansatz für die postulierte präventive Funktion von Selen gegenüber Darmkrebs ist die gastrointestinale Glutathionperoxidase (GPx2), die physiologisch in den proliferierenden Zellen des Kryptengrunds exprimiert wird. Die Regulation dieses Enzyms durch den Wnt-Signalweg, der ebenfalls in proliferierenden Zellen aktiv ist, konnte mittels Reportergenanalyse und endogen auf mRNA- und Proteinebene in Zellkultur gezeigt werden. Die Aktivierung verkürzter Promotorkonstrukte und die Mutation eines potentiellen Bindeelements identifizierten den für die Bindung von TCF und β-Catenin verantwortlichen Bereich. Als Zielgen des Wnt-Signalwegs scheint GPx2 zu den an Proliferationsprozessen beteiligten Genen zu gehören, was unter physiologischen Bedingungen die Aufrechterhaltung des intestinalen Epithels gewährleistet. Bei der Entstehung intestinaler Tumore, die in der Initiationsphase zu über 90 % mit einer konstitutiven Aktivierung des Wnt-Signalwegs einhergeht, wirkt GPx2 möglicherweise prokanzerogen. Die genaue Funktion von GPx2 während der Kanzerogenese bleibt weiter zu untersuchen.
Mercaptursäure und Nukleosidaddukt im Harn als Biomarker in 1-Hydroxymethylpyren-exponierten Ratten
(2002)
1-Methylpyren (MP) ist hepatokanzerogen in neugeborenen männlichen Mäusen. Durch Hydroxylierung an der benzylischen Stelle und anschließende Sulfonierung wird MP zu DNA-reaktivem 1-Sulfooxymethylpyren (SMP) aktiviert. In der Ratte führt die Exposition des benzylischen Alkohols, 1-Hydroxymethylpyren (HMP), zur DNA-Adduktbildung in verschiedenen Geweben. Eventuelle Konsequenz der Toxifizierung ist die Ausscheidung entsprechender Mercaptursäure und Nukleosidaddukt im Harn, welche aufgrund ihrer Herkunft als Biomarker eignen könnten. In dieser Arbeit wird die Ausscheidung der Mercaptursäure und des N2-Desoxyguanosinadduktes in HMP-exponierten Ratten untersucht. Nach der Applikation von HMP bzw. MP wurden weniger als 1 % der Dosis als MPMA über Urin und Faeces ausgeschieden (0 - 48 h). Die Ausscheidung erfolgt hauptsächlich in den ersten 24 h nach der Applikation. MPdG konnte weder in Urin noch in Faeces der HMP-behandelten Tieren identifiziert werden. Nach direkter SMP-Applikation wurde MPdG nur in sehr geringe Menge (weniger als 0,9 ppm in 12 h) im Urin gefunden. Aufgrund der geringen Menge eignet sich MPdG nicht als Biomarker. MPMA dagegen, lässt sich analytisch gut erfassen. Es sollte daher untersucht werden, ob MPMA die Toxifizierung des HMP wiederspiegelt. Die Voraussetzung dafür ist die Kenntnisse über das Metabolismusmuster von HMP. Es wurde daher umfassende Untersuchungen zum Metabolismus des HMP durchgeführt. Die Ergebnisse zeigten, dass mehr als 80 % der Metaboiten in ihrer oxidierten Form (PCS, deren Glucuronsäure-Konjugate sowie phenolische Sulfatester der PCS) ausgeschieden wurden. Demnach spielt die Oxidation des HMP zu PCS eine sehr wichtige Rolle bei der Detoxifizierung und Ausscheidung von HMP. Ferne konnte nachgewiesen werden, dass die Enzyme Alkohol- und Aldehyd-Dehydrogenase an der Oxidation von HMP beteiligt waren. Die Inhibitoren Disulfiram und Ethanol der o. g. Enzyme wurde daher zur Modulation der Detoxifizierung in vivo eingesetzt. Die Veränderungen in der Toxifizierung von HMP zu SMP wurden durch die SMP-Konzentration im Plasma, die DNA-Addukthäufigkeit und die MPMA-Ausscheidung erfasst. Die Vorbehandlung von Disulfiram und Ethanol führte zu tendentielle Erhöhung der SMP-Konzentration im Plasma, DNA-Addukthäufigkeit in der Leber und die MPMA-Ausscheidung. Bemerkenswert ist jedoch, dass bereits eine Dosis von 0,2 g Ethanol/kg Körpermasse bereits zu statistisch signifikanten Erhöhungen der MPMA-Ausscheidung bei weiblichen Ratten.