Rheumatoide Arthritis und Arteriosklerose : Bedeutung der zu Grunde liegenden chronisch entzündlichen Prozesse für die endotheliale Dysfunktion ; Wirkung der antiinflammatorischen Substanz Galiellalacton in murinen Arteriosklerosemodellen

Erhöhte arteriosklerotische und thrombotische Vorfälle sind ein Hauptgrund für die gesteigerten Zahlen kardiovaskulärer Todesfälle von Patienten mit chronisch entzündlichen Erkrankungen wie der rheumatoiden Arthritis (RA). Diese erhöhte Mortalität ist nicht auf die traditionellen Risikofaktoren, wie Alter, Geschlecht, Bluthochdruck oder Diabetes zurückzuführen. Man nimmt an, dass die systemische Entzündung einen nicht-traditionellen Risikofaktor für die erhöhten kardiovaskulären Todesfälle von RA-Patienten darstellt. Da die derzeitige Behandlung der RA zum Teil schwere Nebenwirkungen zur Folge haben kann, war es das Ziel dieser Doktorarbeit, die Zusammenhänge zwischen RA und Arteriosklerose (AS) näher zu untersuchen, sowie die neue antiinflammatorische Substanz Galiellalacton (Gal) für die Behandlung der AS zu charakterisieren.rnIn dem chronisch inflammatorischen Tiermodell der TTP-defizienten Mäuse, dessenrnPhänotyp dem einer humanen RA-Erkrankung ähnelt, konnte eine verschlechterternEndothelfunktion, die als ein erstes Symptom einer erworbenen AS gilt, nachgewiesen werden. Dies konnte auf eine erhöhte Stabilität der Nox2-mRNA zurückgeführt werden, die unabhängig von der erhöhten Expression des Entzündungsmarkers TNFα war. Diese gesteigerte Nox2-Menge führte wiederum zu einer erhöhten Bildung von reaktiven Sauerstoff- und Stickstoffspezies und somit zu einer verringerten Menge an bioaktivem Stickstoffmonoxid, welches die endotheliale Dysfunktion (eDF) bedingte.rnAls ein traditioneller Risikofaktor für das Auftreten von kardiovaskulären Ereignissen gilt unter anderem eine Diabeteserkrankung. Durch die Ausbildung einer Nitrattoleranz bei der Therapie mit organischen Nitraten wie NTG, ISMN oder ISDN kommt es zu der Entwicklung einer eDF. PETN, ein weiteres organisches Nitrat zeigt diese Nebenwirkung nicht. PETN, vermittelt seinen antioxidativen Effekt über die Nrf2-abhängige Induktion der HO-1-Promotoraktivität.rnDie Behandlung von arteriosklerotischen Mäusen (ApoE-/-- und ApoE-/-TFPI+/--Mäuse) mit dem antiinflammatorischen Pilzsekundärmetaboliten Gal zeigte eine verringerte mRNA-Expression von arteriosklerotischen und inflammatorischen Mediatoren, sowie eine reduzierte Thrombenbildung durch eine verringerte Plättchenadhäsion.rnZusammenfassend konnte gezeigt werden, dass inflammationsabhängiger oxidativerrnStress ein Hauptgrund für die entzündungsgetriebene Artheriogenese ist und Galrneine neue Leitsubstanz für die Behandlung dieser Erkrankung ist.

Implementation of enhanced biological phosphorus removal (ebpr) wastewater treatment processes enriched with different microbial communities

The EBPR (Enhanced Biological Phosphorus Removal) is a type of secondary treatment in WWTPs (WasteWater Treatment Plants), quite largely used in full-scale plants worldwide. The phosphorus occurring in aquatic systems in high amounts can cause eutrophication and consequently the death of fauna and flora. A specific biomass is used in order to remove the phosphorus, the so-called PAOs (Polyphosphate Accumulating Organisms) that accumulate the phosphorus in form of polyphosphate in their cells. Some of these organisms, the so-called DPAO (Denitrifying Polyphosphate Accumulating Organisms) use as electron acceptor the nitrate or nitrite, contributing in this way also to the removal of these compounds from the wastewater, but there could be side reactions leading to the formation of nitrous oxides. The aim of this project was to simulate in laboratory scale a EBPR, acclimatizing and enriching the specialized biomass. Two bioreactors were operated as Sequencing Batch Reactors, one enriched in Accumulibacter, the other in Tetrasphaera (both PAOs): Tetrasphaera microorganisms are able to uptake aminoacids as carbon source, Accumulibacter uptake organic carbon (volatile fatty acids, VFA). In order to measure the removal of COD, phosphorus and nitrogen-derivate compounds, different analysis were performed: spectrophotometric measure of phosphorus, nitrate, nitrite and ammonia concentrations, TOC (Total Organic Carbon, measuring the carbon consumption), VFA via HPLC (High Performance Liquid Chromatography), total and volatile suspended solids following standard methods APHA, qualitative microorganism population via FISH (Fluorescence In Situ Hybridization). Batch test were also performed to monitor the NOx production. Both specialized populations accumulated as a result of SBR operations; however, Accumulibacter were found to uptake phosphates at higher extents. Both populations were able to remove efficiently nitrates and organic compounds occurring in the feeding. The experimental work was carried out at FCT of Universidade Nova de Lisboa (FCT-UNL) from February to July 2014.

Thermoelektrische und strukturelle Eigenschaften gefüllter Skutterudite, sowie Verbesserung ihres Gütefaktors durch Nanostrukturierung

Gefüllte Skutterudite mit der Summenformel MxCo4Sb12 sind vielversprechende thermoelektrische Materialien. Die Standardsynthese führt jedoch oft zur Bildung von MSbx, Sb, CoSb oder CoSb2 als Nebenphasen. In dieser Arbeit wird eine neue zweistufige Synthese vorgestellt, bei der die Bildung des Kieftits (CoSb3) getrennt von dem topotaktischen Füllen mit dem Metallatom M erfolgt. Dieser Ansatz erlaubt eine Durchführung der Reaktion bei niedrigeren Temperaturen mit kürzeren Reaktionszeiten. Ein geringer Antimon-Unterschuss im so erhaltenen Kieftit erhöht die Anzahl der Ladungsträger und unterdrückt die Bildung von Verunreinigungsphasen. Zunächst wurden Skutteruditproben mit der nominellen Zusammensetzung InxCo4Sb12 mit x = 0,12; 0,15; 0,18 und 0,20 in hoher Reinheit hergestellt und mit Spark Plasma Sintering (SPS) kompaktiert. Messaufnahmen mit Potential- und Seebeck-Mikrosonde und Rasterelektronenmikroskop zeigten eine hohe Probenhomogenität. Produkte waren nahezu phasenrein, was eine Untersuchung der Transporteigenschaften ohne Verfälschung durch Nebenphasen ermöglichte. Die quantitative Phasenanalyse mittels Synchrotron-Beugungsdaten zeigte < 0,1 % InSb bei In0,18Co4Sb12 und In0,20Co4Sb12, sowie eine lineare Korrelation zwischen dem wahren Füllgrad und der Gitterkonstante. Die Bindung von < 0,1 % InSb verringerte den Füllgrad der nominellen In0,20Co4Sb12-Probe auf x = 0,144. Die nominelle In0,18Co4Sb12-Probe mit dem wahren Gehalt x = 0,160 hatte den höchsten zT-Wert nahe eins bei 420 °C. Es konnte anschließend die Anwendbarkeit der Synthesemethode für Barium- und mehrfach gefüllte (Na+In) Skutterudite gezeigt werden. Die Na-gefüllte Probe war gegenüber der thermischen Behandlung in der SPS oder der Charakterisierung instabil. Alle Verbindungen wurden gesintert und ihre Transporteigenschaften wurden charakterisiert. Des weiterem wurde der Einfluss der Konzentration der Korngrenzen bei den Mischungen von zu Nanomaßstab vermahlenem In0,18Co4Sb12 (Partikelgrößen zwischen 20 und 100 nm) mit dem ursprünglichen Bulk untersucht. Proben mit verschiedenen Anteilen von Nanopulver wurden gesintert, ihre thermoelektrischen und strukturellen Eigenschaften wurden untersucht. Die Gütezahl zT von 1,39 bei 375 °C wurde bei der Probe mit gleichen Anteilen des Nano- und des unbehandelten Pulvers erreicht. Die Komposite mit Anteilen <10 % oder >75 % des Nanopulvers zeigten keine Verbesserung gegenüber der unbehandelten Verbindung.rn

The role of DNA damage in the pathogenesis of nitrate tolerance - nitrosative versus oxidative DNA damage

6. Summary Despite the lack of direct evidence from large clinical trials for mutagenic and genotoxic effects of GTN therapy, the present study show s the induction of pre-mutagenic lesions, such as 8- oxo - G and O 6 - me - G by GTN t reatment as well as increased formation of DNA strand breaks. These results were obtained in an in vitro (EA.hy 926 – human endothelial cell line) and in vivo (Wistar rats and C57BL/6 mice) setting. However, GTN - induced DNA damage had no effect on the degr ee of nitrate tolerance but only on other pathological side effects such as oxidative stress, as confirmed by studies in MGMT knockout mice. Of clinical importance , this study establishes potent apoptotic properties of organic nitrates, which has been demo nstrated by the levels of the novel apoptotic marker and caspase - 3 substrate, fractin, as well as levels of cleaved caspase - 3 , the activated form of this pro - apoptotic enzyme . The p rotein analy tical data ha ve been confirmed by an independent assay for the apoptosis , Cell death detection assay (TUNEL) . First, these GTN - mediated apoptotic effects may account for the previously reported anti - cancer effects of GTN therapy (probably based on induction of apoptosis in tumor cells). Second, these GTN - mediated apop totic effects may account for the increased mortality rates observed in the group of organic nitrate - treated patients as reported by two independent meta - analysis (probably due to induction of apoptosis in highly beneficial endothelial progenitor cells as well as in cardiomyocytes during wound healing and cardiac remodeling) . Summary of the current investigations can be seen in Figure 18.

GSR deposition along the bullet path in contact shots to composite models

In contact shots, all the materials emerging from the muzzle (combustion gases, soot, powder grains, and metals from the primer) will be driven into the depth of the entrance wound and the following sections of the bullet track. The so-called "pocket" ("powder cavity") under the skin containing soot and gunpowder particles is regarded as a significant indicator of a contact entrance wound since one would expect that the quantity of GSR deposited along the bullet's path rapidly declines towards the exit hole. Nevertheless, experience has shown that soot, powder particles, and carboxyhemoglobin may be found not only in the initial part of the wound channel, but also far away from the entrance and even at the exit. In order to investigate the propagation of GSRs under standardized conditions, contact test shots were fired against composite models of pig skin and 25-cm-long gelatin blocks using 9-mm Luger pistol cartridges with two different primers (Sinoxid® and Sintox®). Subsequently, 1-cm-thick layers of the gelatin blocks were examined as to their primer element contents (lead, barium, and antimony as discharge residues of Sinoxid® as well as zinc and titanium from Sintox®) by means of X-ray fluorescence spectroscopy. As expected, the highest element concentrations were found in the initial parts of the bullet tracks, but also the distal sections contained detectable amounts of the respective primer elements. The same was true for amorphous soot and unburned/partly burned powder particles, which could be demonstrated even at the exit site. With the help of a high-speed motion camera it was shown that for a short time the temporary cavitation extends from the entrance to the exit thus facilitating the unlimited spread of discharge residues along the whole bullet path.

Geochemical Modeling and Analysis of the Frac Water Used in the Hydraulic Fracturing of the Marcellus Formation, Pennsylvania

The hydraulic fracturing of the Marcellus Formation creates a byproduct known as frac water. Five frac water samples were collected in Bradford County, PA. Inorganic chemical analysis, field parameters analysis, alkalinity titrations, total dissolved solids(TDS), total suspended solids (TSS), biological oxygen demand (BOD), and chemical oxygen demand (COD) were conducted on each sample to characterize frac water. A database of frac water chemistry results from across the state of Pennsylvania from multiple sources was compiled in order to provide the public and research communitywith an accurate characterization of frac water. Four geochemical models were created to model the reactions between frac water and the Marcellus Formation, Purcell Limestone, and the oil field brines presumed present in the formations. The average concentrations of chloride and TDS in the five frac water samples were 1.1 �± 0.5 x 105 mg/L (5.5X average seawater) and 140,000 mg/L (4X average seawater). BOD values for frac water immediately upon flow back were over 10X greater than the BOD of typical wastewater, but decreased into the range of typical wastewater after a short period of time. The COD of frac water decreases dramatically with an increase in elapsed time from flow back, but remain considerably higher than typicalwastewater. Different alkalinity calculation methods produced a range of alkalinity values for frac water: this result is most likely due to high concentrations of aliphatic acid anions present in the samples. Laboratory analyses indicate that the frac watercomposition is quite variable depending on the companies from which the water was collected, the geology of the local area, and number of fracturing jobs in which the frac water was used, but will require more treatment than typical wastewater regardless of theprecise composition of each sample. The geochemical models created suggest that the presence of organic complexes in an oil field brine and Marcellus Formation aid in the dissolution of ions such as bariumand strontium into the solution. Although equilibration reactions between the Marcellus Formation and the slickwater account for some of the final frac water composition, the predominant control of frac water composition appears to be the ratio of the mixture between the oil field brine and slickwater. The high concentration of barium in the frac water is likely due to the abundance of barite nodules in the Purcell Limestone, and the lack of sulfate in the frac water samples is due to the reducing, anoxic conditions in the earth's subsurface that allow for the degassing of H2S(g).

Ground Water Impact Assessment in Rural Areas of North-Eastern Part of Ukrain

Protecting ground water from pollution in the rural areas of Ukraine is a priority, particularly because the largely unconfined aquifers are used by local people for potable water supply. The most important aim of this project was to characterise the scale and impact of pollution sources on groundwater quality according to the nature of groundwater vulnerability and of the particular sources of pollution. Analysis of 50 different maps of Ukraine according to geochemical, natural and landscape features, together with field studies made it possible to identify areas with different pollution risk factors. The most harmful sources of groundwater contamination providing nitrates, petrol, heavy metals and other pollutants were identified and an integral method of groundwater quality assessment was worked out. The main sources of groundwater contamination in rural areas of Ukraine were identified and listed in order of importance. Magmedov also puts forward recommendations for improving monitoring of groundwater quality as an essential part of sustainable development in rural areas of Ukraine.

A dedicated animal model for mechanical thrombectomy in acute stroke

BACKGROUND: Recent studies have focused on mechanical thrombectomy as a means to reduce the time required for revascularization and increase the revascularization rate in acute stroke. To date no systematic evaluation has been made of the different mechanical devices in this novel and fast-developing field of endovascular interventions. To facilitate such evaluations, we developed a specific in vivo model for mechanical thrombectomy that allows visualization of dislocation or fragmentation of the thrombus during angiographic manipulation. METHODS: Angiography and embolization with a preformed thrombus was performed in 8 swine. The thrombus was generated by mixing 25 IU bovine thrombin and 10 mL autologous blood. For visualization during angiography, 1 g barium sulfate was added. RESULTS: The preformed thrombus exhibited mechanical stability, reproducibility, and high radiographic absorption, providing excellent visibility during angiography. The setting allowed selective embolization of targeted vessels without thrombus fragmentation. Despite the application of barium sulfate no local or systemic reaction occurred. Histologic evaluation revealed no intimal damage caused by the thrombus or contrast agent washout. CONCLUSION: The model presented here allows selective and reliable thromboembolization of vessels that reproduce the anatomic and hemodynamic situation in acute cerebrovascular stroke. It permits visualization of the thrombus during angiography and intervention, providing unique insight into the behavior of both thrombus and device, which is potentially useful in the development and evaluation of mechanical clot retrieval in acute cerebrovascular stroke.

Electroanatomic reconstruction of the left atrium, pulmonary veins, and esophagus compared with the "true anatomy" on multislice computed tomography in patients undergoing catheter ablation of atrial fibrillation

BACKGROUND: Current concepts of catheter ablation for atrial fibrillation (AF) commonly use three-dimensional (3D) reconstructions of the left atrium (LA) for orientation, catheter navigation, and ablation line placement. OBJECTIVES: The purpose of this study was to compare the 3D electroanatomic reconstruction (Carto) of the LA, pulmonary veins (PVs), and esophagus with the true anatomy displayed on multislice computed tomography (CT). METHODS: In this prospective study, 100 patients undergoing AF catheter ablation underwent contrast-enhanced spiral CT scan with barium swallow and subsequent multiplanar and 3D reconstructions. Using Carto, circumferential plus linear LA lesions were placed. The esophagus was tagged and integrated into the Carto map. RESULTS: Compared with the true anatomy on CT, the electroanatomic reconstruction accurately displayed the true distance between the lower PVs; the distances between left upper PV, left lower PV, right lower PV, and center of the esophagus; the longitudinal diameter of the encircling line around the funnel of the left PVs; and the length of the mitral isthmus line. Only the distances between the upper PVs, the distance between the right upper PV and esophagus, and the diameter of the right encircling line were significantly shorter on the electroanatomic reconstructions. Furthermore, electroanatomic tagging of the esophagus reliably visualized the true anatomic relationship to the LA. On multiple tagging and repeated CT scans, the LA and esophagus showed a stable anatomic relationship, without relevant sideward shifting of the esophagus. CONCLUSION: Electroanatomic reconstruction can display with high accuracy the true 3D anatomy of the LA and PVs in most of the regions of interest for AF catheter ablation. In addition, Carto was able to visualize the true anatomic relationship between the esophagus and LA. Both structures showed a stable anatomic relationship on Carto and CT without relevant sideward shifting of the esophagus.

Dysphagia due to triple A syndrome: successful treatment of achalasia by balloon dilatation

Triple A syndrome is a rare autosomal recessive inherited disorder which is characterized by alacrima, adrenal insufficiency, and achalasia. We report on a 14-year old girl with dysphagia, regurgitation, and vomiting since 5 years. At the age of five years an Addison crisis was diagnosed and cortisone substitution was initiated. In addition, the patient had episodes of conjunctivitis. Severe esophagitis and candida infection were diagnosed by esophago-gastro-duodenoscopy and treated with omeprazole and fluconazole. The esophageal barium swallow was typical for achalasia. Medical treatment of achalasia with oral nifedipine resulted only in a partial and temporal improvement. But after seven balloon dilatations dysphagia and nocturnal coughing improved clearly and a remarkable gain of weight could be seen. Direct sequencing showed a homozygous nonsense mutation in exon 11 of the AAAS gene leading to truncation at position 342 of the 546 amino acid protein. CONCLUSION: Triple A syndrome has to be considered in patients with dysphagia. In our patient, the absence of tears since birth followed by adrenal insufficiency were early signs of the triple A syndrome. Balloon dilatation of the esophago-gastric junction is an effective treatment, which can avoid surgical interventions.

Synthesis of PDMS-metal oxide hybrid nanocomposites using an in situ sol-gel route

Organic-inorganic hybrid nanocomposites are widely studied and applied in broad areas because of their ability to combine the flexibility, low density of the organic materials with the hardness, strength, thermal stability, good optical and electronic properties of the inorganic materials. Polydimethylsiloxane (PDMS) due to its excellent elasticity, transparency, and biocompatibility has been extensively employed as the organic host matrix for nanocomposites. For the inorganic component, titanium dioxide and barium titanate are broadly explored as they possess outstanding physical, optical and electronic properties. In our experiment, PDMS-TiO2 and PDMS-BaTiO3 hybrid nanocomposites were fabricated based on in-situ sol-gel technique. By changing the amount of metal precursors, transparent and homogeneous PDMS-TiO2 and PDMS-BaTiO3 hybrid films with various compositions were obtained. Two structural models of these two types of hybrids were stated and verified by the results of characterization. The structures of the hybrid films were examined by a conjunction of FTIR and FTRaman. The morphologies of the cross-sectional areas of the films were characterized by FESEM. An Ellipsometer and an automatic capacitance meter were utilized to evaluate the refractive index and dielectric constant of these composites respectively. A simultaneous DSC/TGA instrument was applied to measure the thermal properties. For PDMS-TiO2 hybrids, the higher the ratio of titanium precursor added, the higher the refractive index and the dielectric constant of the composites are. The highest values achieved of refractive index and dielectric constant were 1.74 and 15.5 respectively for sample PDMS-TiO2 (1-6). However, when the ratio of titanium precursor to PDMS was as high as 20 to 1, phase separation occurred as evidenced by SEM images, refractive index and dielectric constant decreased. For PDMS-BaTiO3 hybrids, with the increase of barium and titanium precursors in the system, the refractive index and dielectric constant of the composites increased. The highest value was attained in sample PDMS-BaTiO3 (1-6) with a refractive index of 1.6 and a dielectric constant of 12.2. However, phase separation appeared in SEM images for sample PDMS-BaTiO3 (1-8), the refractive index and dielectric constant reduced to lower values. Different compositions of PDMS-TiO2 and PDMS-BaTiO3 hybrid films were annealed at 60 °C and 100 °C, the influences on the refractive index, dielectric constant, and thermal properties were investigated.

Wetting and Reactive Air Brazing of BSCF for Oxygen Separation Devices

The goals of this project are to develop a Reactive Air Brazing (RAB) alloy and process for joining Barium strontium cobalt ferrite (BSCF), and to develop a fundamental understanding of the wettability and microstructral development due to reaction kinetics in BSCF/Ag-MexOy systems.

A Study of the Action of Various Energizers in the Carburization of Steel

The object of casehardening is to produce articles of steel having a tough or ductile interior and a hard sur­face. Quenching produces a surface somewhat harder than the interior, but in order to obtain a high surface hard­ness, the percentage of alloying elements in the steel must be raised to such an extent that the core or central por­tion becomes hard and brittle also.

Tamoxifen inhibits TRPV6 activity via estrogen receptor-independent pathways in TRPV6-expressing MCF-7 breast cancer cells

The epithelial calcium channel TRPV6 is upregulated in breast carcinoma compared with normal mammary gland tissue. The selective estrogen receptor modulator tamoxifen is widely used in breast cancer therapy. Previously, we showed that tamoxifen inhibits calcium uptake in TRPV6-transfected Xenopus oocytes. In this study, we examined the effect of tamoxifen on TRPV6 function and intracellular calcium homeostasis in MCF-7 breast cancer cells transiently transfected with EYFP-C1-TRPV6. TRPV6 activity was measured with fluorescence microscopy using Fura-2. The basal calcium level was higher in transfected cells compared with nontransfected cells in calcium-containing solution but not in nominally calcium-free buffer. Basal influxes of calcium and barium were also increased. In transfected cells, 10 mumol/L tamoxifen reduced the basal intracellular calcium concentration to the basal calcium level of nontransfected cells. Tamoxifen decreased the transport rates of calcium and barium in transfected cells by 50%. This inhibitory effect was not blocked by the estrogen receptor antagonist, ICI 182,720. Similarly, a tamoxifen-induced inhibitory effect was also observed in MDA-MB-231 estrogen receptor-negative cells. The effect of tamoxifen was completely blocked by activation of protein kinase C. Inhibiting protein kinase C with calphostin C decreased TRPV6 activity but did not alter the effect of tamoxifen. These findings illustrate how tamoxifen might be effective in estrogen receptor-negative breast carcinomas and suggest that the therapeutic effect of tamoxifen and protein kinase C inhibitors used in breast cancer therapy might involve TRPV6-mediated calcium entry. This study highlights a possible role of TRPV6 as therapeutic target in breast cancer therapy.

Dating groundwater in the Bohemian Cretaceous Basin: Understanding tracer variations in the subsurface

The northern section of the Bohemian Cretaceous Basin has been the site of intensive U exploitation with harmful impacts on groundwater quality. The understanding of groundwater flow and age distribution is crucial for the prediction of the future dispersion and impact of the contamination. State of the art tracer methods (3H, 3He, 4He, 85Kr, 39Ar and 14C) were, therefore, used to obtain insights to ageing and mixing processes of groundwater along a north–south flow line in the centre of the two most important aquifers of Cenomanian and middle Turonian age. Dating of groundwater is particularly complex in this area as: (i) groundwater in the Cenomanian aquifer is locally affected by fluxes of geogenic and biogenic gases (e.g. CO2, CH4, He) and by fossil brines in basement rocks rich in Cl and SO4; (ii) a thick unsaturated zone overlays the Turonian aquifer; (iii) a periglacial climate and permafrost conditions prevailed during the Last Glacial Maximum (LGM), and iv) the wells are mostly screened over large depth intervals. Large disagreements in 85Kr and 3H/3He ages indicate that processes other than ageing have affected the tracer data in the Turonian aquifer. Mixing with older waters (>50 a) was confirmed by 39Ar activities. An inverse modelling approach, which included time lags for tracer transport throughout the unsaturated zone and degassing of 3He, was used to estimate the age of groundwater. Best fits between model and field results were obtained for mean residence times varying from modern up to a few hundred years. The presence of modern water in this aquifer is correlated with the occurrence of elevated pollution (e.g. nitrates). An increase of reactive geochemical indicators (e.g. Na) and radiogenic 4He, and a decrease in 14C along the flow direction confirmed groundwater ageing in the deeper confined Cenomanian aquifer. Radiocarbon ages varied from a few hundred years to more than 20 ka. Initial 14C activity for radiocarbon dating was calibrated by means of 39Ar measurements. The 14C age of a sample recharged during the LGM was further confirmed by depleted stable isotope signatures and near freezing point noble gas temperature. Radiogenic 4He accumulated in groundwater with concentrations increasing linearly with 14C ages. This enabled the use of 4He to validate the dating range of 14C and extend it to other parts of this aquifer. In the proximity of faults, 39Ar in excess of modern concentrations and 14C dead CO2 sources, elevated 3He/4He ratios and volcanic activity in Oligocene to Quaternary demonstrate the influence of gas of deeper origin and impeded the application of 4He, 39Ar and 14C for groundwater dating.