Temporal and Spatial Variations of Ions, Isotopes and Agricultural Contaminants in Surface Waters and Groundwater of Nebraska's Rainwater Basin Wetland Region

The wetlands of south-central Nebraska’s Rainwater Basin region are considered of international importance as a habitat for millions of migratory birds, but are being endangered by agricultural practices. The Rainwater Basin extends across 17 counties and covers 4,000 square miles. The purpose of this study was to assemble baseline chemical data for several representative wetlands across the Rainwater Basin region, and determine the use of these chemical data for investigating groundwater recharge. Eight representative wetlands were chosen across the Rainwater Basin to monitor surface and groundwater chemistry. At each site, a shallow well and deep well were installed and sampled once in the summer of 2009 and again in the spring of 2010. Wetland surface water was sampled monthly from April, 2009 to May, 2010. Waters were analyzed for major ions, nutrients, pesticides and oxygen-18 and deuterium isotopes at the University of Nebraska Water Sciences Laboratory. Geochemical analysis of surface waters presents a range of temporal and spatial variations. Wetlands had variable water volumes, isotopic compositions, ion chemistries and agricultural contaminant levels throughout the year and, except for a few trends, theses variations cannot be predicted with certainty year-to-year or wetland-to-wetland. Isotopic compositions showed evaporation was a contributor to water loss, and thus, did impact water chemistry. Surface water nitrate concentrations ranged from <0.10 to 4.04 mg/L. The nitrate levels are much higher in the groundwater, ranging from <0.10 to 18.4 mg/L, and are of concern because they are found above the maximum contaminant level (MCL) of 10 mg/L. Atrazine concentrations in surface waters ranged from <0.05 to 10.3 ppb. Groundwater atrazine concentrations ranged from <0.05 to 0.28 ppb. The high atrazine concentrations in surface waters are of concern as they are above the MCL of 3 ppb, and the highest levels occur during the spring bird migration. Most sampled groundwaters had detectable tritium indicating a mix of modern (<5 to 10 years old) and submodern (older than 1950s) recharge. The groundwater also had differences in chemical and isotope composition, and in some cases, increased nitrate concentrations, between the two sampling periods. Modern groundwater tritium ages and changes in groundwater chemical and isotopic compositions may indicate connections with surface waters in the Rainwater Basin.

Isotopic, trace element and nutrient characterization of coastal waters from Ubatuba inner shelf area, south-eastern Brazil

Stable isotopes, tritium, radium isotopes, radon, trace elements and nutrients data were collected during two sampling campaigns in the Ubatuba coastal area (south-eastern Brazil) with the aim of investigating submarine groundwater discharge (SGD) in the region. The isotopic composition (delta D, delta(18)O, (3)H) of submarine waters was characterised by significant variability and heavy isotope enrichment. The stable isotopes and tritium data showed good separation of groundwater and seawater groups. The contribution of groundwater in submarine waters varied from a few % to 17%. Spatial distribution of (222)Rn activity concentration in surface seawater revealed changes between 50 and 200 Bq m(-3) which were in opposite relationship with observed salinities. Time series measurements of (222)Rn activity concentration in Flamengo Bay (from 1 to 5 kBq m(-3)), obtained by in situ underwater gamma-spectrometry showed a negative correlation between the (222)Rn activity concentration and tide/salinity. This may be caused by sea level changes as tide effects induce variations of hydraulic gradients, which increase (222)Rn concentration during lower sea level, and opposite, during high tides where the (222)Rn activity concentration is smaller. The estimated SGD fluxes varied during 22-26 November between 8 and 40 cm d(-1), with an average value of 21 cm d(-1) (the unit is cm(3)/cm(2) per day). The radium isotopes and nutrient data showed scattered distributions with offshore distance and salinity. which implies that in a complex coast with many small bays and islands, the area has been influenced by local currents and groundwater-seawater mixing. SGD in the Ubatuba area is fed by coastal contaminated groundwater and re-circulated seawater (with small admixtures of groundwater). which claims for potential environmental concern with implications on the management of freshwater resources in the region. (C) 2007 Elsevier Ltd. All rights reserved.

WATER HOMEOSTASIS, FRAILTY AND COGNITIVE FUNCTION IN THE NURSING HOME

Objective: - To develop and test a practical clinical method to assess frailty in nursing homes; - To investigate the relationship between cognitive status of the elderly and the balance between water compartments of their body composition. Design and subjects: Cross-sectional study, conducted at two nursing homes in Boston-MA. Methods: Body mass and height (Ht) were evaluated to calculate BMI (body mass index, in Kg/m(2)). The cognitive decline was evaluated based on the scores obtained from the Mini-Mental State Examination (MMSE); The extracellular to total body water ratio (ECW/TBW) was calculated after the analysis of TBW from deuterium and tritium dilution and ECW from bromide dilution. Single-frequency BIA analysis data were investigated for resistance (R) and reactance (Xc), plotted in an R/Ht Xc/Ht graph (vectorial analysis-BIVA). The BIVA results of nursing home residents were compared against the data obtained from the NHANES Ill study. TBW and ECW values were compared with a group of free-living elderly volunteers. Results: The ECW/TBW was significantly higher in nursing home residents than in the free-living individuals. BIVA analysis showed significantly higher Xc/Ht values in the reference subjects. The MMSE did not present a significant correlation with ECW/TBW for either gender. Conclusion: We proposed the ECW/TBW ratio and BIVA as surrogate methods for the clinical assessment of frailty. We tested successfully both approaches with nursing home patients and free-living volunteers and compared them to a national data base. The advent of new, portable instruments will enable field tests to further validate our proposed "Frailty Factor" in future studies. We found no correlation between frailty and cognitive decline in the nursing home.

Mobilisation von Schwermetallen im Bereich rheinhessisches Hügelland als Folge anthropogen bedingten Nitrateintrages - Folgen für den Grundwasserschutz ?

Kurzfassung Zielsetzung der vorliegenden Arbeit war eine hydrogeologische Untersuchung zur Herkunftsbestimmung der Grundwässer im Raum Alzey/Rheinhessen. Ein erster Anlaß zu weiteren Untersuchungen war durch erhöhte Schwermetallgehalte in den Rohwässern gegeben. Im Alzeyer Raum können Kluftgrundwasserleiter (Rotliegendes und z.T. verkarstetes Kalktertiär) sowie Porengrundwasserleiter (Tertiäre Meeressande und Ablagerungen des Quartärs) unterschieden werden. Aufgrund der Lösungsinhalte ergibt sich eine Typisierung in erdalkalische Hydrogenkarbonatwässer bzw. Austauschwässer, bei denen Erdalkalien durch Alkalien ersetzt wurden. Mit verschiedenen Grundwassermodellierungen konnten mögliche 'Grundwasserneubildungs-Pfade' aufgezeigt werden. Die 1996 durchgeführte Herabsetzung des Arsengrenzwertes zwang viele Versorgungsunternehmen weitere Verfahrenstechniken zur Grundwasseraufbereitung anzuwenden. Es zeigten sich erhebliche Schwierigkeiten bei der Umsetzung der EU-Wasserrahmenrichtlinie bezogen auf das lokale Untersuchungsgebiet Alzey. Schwermetallmobilisationen im Untergrund können aufgrund anthropogen eingetragener Nitrate im sonst reduzierenden Milieu ermöglicht werden. Mittels Tritium- und FCKW-Analysen konnten Alter bzw. Verweilzeiten der Wässer bestimmt werden. Zusammen mit den Werten von Deuterium und Sauerstoff-18, Schwefel-34 und Sauerstoff-18 aus dem im Wasser gelösten Sulfat und Stickstoff-15 und Sauerstoff-18 aus dem im Wasser gelösten Nitrat, konnten Aussagen über Änderungen im Aquifermilieu getroffen werden. Es zeigte sich, daß selbst bei einem völligen Stop des Düngemitteleintrages, die Schwermetallmobilisationen in Grundwässern im Bereich Rheinhessisches Hügelland innerhalb der nächsten 30 Jahre nicht abnehmen werden.

Untersuchung systematischer Effekte und erste Tritiummessungen mit dem verbesserten Mainzer Neutrinomassenexperiment

Das Mainzer Neutrinomassenexperiment untersucht dasBetaspektrum von Tritium in der Naehe des Endpunktes undbestimmt daraus die Ruhemasse des Elektronantineutrinos. DieMessungen werden mit einem integrierenden, elektrostatischenGegenfeldspektrometer durchgefuehrt, bei dem das Prinzip dermagnetischen adiabatischen Kollimation angewandt wird, umgleichzeitig hohe Raumwinkelakzeptanz (bis maximal 2Pi) undeine sehr gute Energieaufloesung (E/dE=4000) zu erreichen.Als Tritiumquellen werden schockkondensierte Tritiumfilmeeingesetzt.Der Schwerpunkt dieser Arbeit bestand darin, das MainzerExperiment in entscheidenden Punkten zu verbessern. Das Zielwar, die Sensitivitaet auf die Neutrinomasse zu erhoehen unddie von der Troitskgruppe berichtete Anomalie imTritium-Betaspektrum zu ueberpruefen. Diese beschreibt einemonoenergetische Line variabler Amplitude einige eVunterhalb des Betaendpunktes, deren Positioin mit einerHalbjahresperiode variiert.Nach dem Umbau 1995-1997 wurde der Endpunktsbereich desTritium-Betaspektrums bis Ende 1998 in 4 Messphasen ueberjeweils 1 Monat vermessen. Die erreichte Sensitivitaet derMainzer Daten war ausreichend, um die Troitsk Hypothese zuueberpruefen. Eine der 3 im Jahr 1998 durchgefuehrtenMessungen unterstuetzt die Hypothese, die Daten einer 6Monate spaeter durchgefuehrten Messung dagegen nicht. Somitstehen die Ergebnisse im Widerspruch zur postuliertenHalbjahresperiode.Die neuen Messungen zeigen bei Vergroesserung desAuswerteintervalls keinen Trend zu unphysikalisch negativenMassenquadraten. Es kann eine neue Neutrinomassenobergerenzevon 3.0 eV/c/c abgeleitet werden.

Sviluppo delle tecnologie del circuito a metallo liquido del modulo di blanket HCLL del reattore a fusione ITER e DEMO

Lo scopo della presente tesi di dottorato è di illustrare il lavoro svolto nella progettazione del circuito a metallo liquido del Test Blanket System (TBS) Helium Cooled Lithium Lead (HCLL), uno dei sistemi fondamentali del reattore sperimentale ITER che dovrà dimostrare la fattibilità di produrre industrialmente energia elettrica da processi di fusione nucleare. Il blanket HCLL costituisce una delle sei configurazioni che verranno testate in ITER, sulla base degli esperimenti condotti nei 10 dieci anni di vita del reattore verrà selezionata la configurazione che determinerà la costituzione del primo reattore dimostrativo per la produzione di un surplus di energia elettrica venti volte superiore all’energia consumata, DEMO. Il circuito ausiliario del blanket HCLL è finalizzato, in DEMO all’estrazione del trizio generato mediante il TES; ed in ITER alla dimostrazione della fattibilità di estrarre il trizio generato e di poter gestire il ciclo del trizio. Lo sviluppo dei componenti, svolto in questa tesi, è accentrato su tale dispositivo, il TES. In tale ambito si inseriscono le attività che sono descritte nei capitoli della seguente tesi di dottorato: selezione e progettazione preliminare del sistema di estrazione del trizio dalla lega eutettica Pb15.7Li del circuito a metallo liquido del TBM HCLL; la progettazione, realizzazione e qualifica dei sensori a permeazione per la misura della concentrazione di trizio nella lega eutettica Pb15.7Li; la qualificazione sperimentale all’interno dell’impianto TRIEX (TRItium EXtarction) della tecnologia selezionata per l’estrazione del trizio dalla lega; la progettazione della diagnostica di misura e controllo del circuito ausiliario del TBM HCLL.

Synthese, Tritierung, 99m Tc-Markierung und Evaluierung von Benzamid-Derivaten zur Visualisierung der D 2, D 3-Rezeptoren

Die Diagnose und Therapie von neurodegenerativen Krankheiten, wie beispielsweise Morbus Parkinson besitzt in der heutigen Gesellschaft eine immer größere Bedeutung. Über moderne, bild¬gebende nuklear¬medizinische Verfahren wie SPECT und PET ist es mit geeigneten Radioliganden möglich, Morbus Parkinson vor dem Auftreten von Symptomen zu diagnostizieren. Ein wichtiger Ansatzpunkt zur Diagnose von Morbus Parkinson ist die Visualisierung der postsynaptischen Dopamin-Rezeptoren über radioaktiv (11C, 18F, 123I) markierte Benz¬amid-Derivate. Auf Grundlage der (S)-Pyrolidin-2,3-dimethoxy-Benzamid-Struktur des 18F-Liganden Fallypride wurden verschiedene 99mTc-markierte Benzamid-Derivate als potentielle Radio¬liganden zur Parkinson-Diagnostik entwickelt. Um das Potential von Metall-konjugierten Benzamiden abschätzen zu können, wurden zunächst einfache Vergleichssubstanzen entwickelt. Diese sollten die Einführung eines Chelators simulieren und wurden hierfür hinsichtlich ihrer in vitro-Bindungsaffinitäten zu den Dopamin-, Serotonin- und adrenergen Rezeptoren evaluiert. Die zunächst entwickelten Derivate mit unterschiedlichen Kettenlängen zur Kopplung des Chelators zeigten für die Propylkette Affinitäten im nanomolaren Bereich. Im Anschluss sollten basierend auf diesen Ergebnissen vier verschiedene Chelatoren (Carbony-Cyclopentadienyl, Amido-Cyclopentadienyl, 2-Pyridyl-Imin und N2S2) über eine Propylkette an die 5-Position der Benzamidgrundstruktur gekoppelt werden. Die geplante Synthese des Carbonyl-Cyclopentadienyl-Derivates gelang jedoch nicht. Für die weiteren Chelatoren (Amido-Cyclopentadieny, 2-Pyridyl-Imin und N2S2) konnten die jeweiligen Markierungs¬vorläufer und Rhenium-Komplexe dargestellt werden, die ebenfalls hinsichtlich ihrer Bindungs¬affinitäten evaluiert wurden. Die erzielten Affinitäten zeigten, dass eine Über¬tragung der Affinitäten der einfachen Vergleichssubstanzen auf die komplexeren Metall-Benzamide nicht möglich war. Insbesondere der N2S2-Rhenium-Komplex besitzt nur noch geringe Affinität (490 - 900 nM) zu den D2- und D3-Rezeptoren. Die mittel-affinen 2-Pyridyl-Imin- und Amdio¬cyclopentadien-Komplexe wurden mit 99mTc markiert und die Markierungsausbeute hinsichtlich Reaktionstemperatur, Markierungs-vorläuferkonzentration und Heizmethoden optimiert. Dabei konnte der Imin-Komplex quantitativ mittels fac-[99mTc(CO)3(H2O)3]+ in 30 Minuten bei 45°C markiert werden. Der Amido-Cyclopentadien-Komplex konnte über die Umsetzung des Ferrocen-Markierungsvorläufer mit Mn(CO)5Br und [99mTcO4]- in Ausbeuten von bis zu 60 % markiert werden. Im Anschluss an die Markierungen wurden die 99mTc-Komplexe über HPLC isoliert und in in vitro-Auto¬radiographien von Rattenhirnschnitten weiter evaluiert. Die erhaltenen Ergebnisse bestätigten die für die Rhenium-Komplexe erzielten Affinitäten und zeigten keine spezifische Anreicherung in bestimmten Hirnarealen. Aus diesen Ergebnissen kann ge¬schlossen werden, dass die dargestellten 99mTc-Benzamide aufgrund mangelnder Affinitäten und einer hohen unspezifischen Bindung keine geeigneten Liganden zur Darstellung der D2- und D3- Rezeptoren sind. Um die dargestellten 99mTc-Benzamide mit [18F]Fallypride vergleichen zu können, wurde zusätzlich [3H]Fallypride dargestellt. Hierfür wurde zunächst der Nor-Markierungsvor¬läufer synthetisiert und die Markierungsausbeute optimiert. Die finale Umsetzung mit [3H]Methylnosylat ergab nach HPLC-Aufreinigung 15 mCi [3H]Fallypride mit einer radio¬chemischen Reinheit von >99,5 %. Erste Autoradiographien zeigten eine hohe Anreicherung des Liganden im Striatum, verbunden mit einer sehr niedrigen unspezifischen Bindung.

Glukose-konjugierte Hemmstoffe als Strategie der gezielten Hemmung des DNA-Reparaturproteins O 6 -Methylguanin-DNA-Methyltransferase in Tumorzellen und der Einfluss von ATP-binding cassette Transportern

Das DNA-Reparaturprotein O6-Methylguanin-DNA-Methyltransferase [MGMT] ist der Hauptresistenzfaktor gegenüber der zytotoxischen Wirkung von SN1-alkylierenden Zytostatika in der Tumortherapie. Die Verwendung der MGMT-Hemmstoffe O6-Benzylguanin [O6BG] und O6-(4-Bromothenyl)guanin [O6BTG] führte zu einer Sensibilisierung des Normalgewebes, was eine Dosis-Reduktion der Zytostatika erforderlich machte und die erhoffte Therapieverbesserung verhinderte. Aus diesem Grund ist eine Strategie der selektiven Hemmung des MGMT-Proteins (Targeting-Strategie) erforderlich, um die systemische Toxizität in der Kombinationsbehandlung zu reduzieren. In dieser Arbeit wurde die Anwendbarkeit der Glukose-Konjugation als Targeting-Strategie untersucht, da Tumorzellen einen erhöhten Glukoseverbrauch aufweisen und demzufolge Glukosetransporter überexprimieren. Die Glukose-Konjugate O6BG-Glu und O6BTG-Glu inhibierten MGMT in Tumorzellen und sensibilisierten die Zellen gegenüber den alkylierenden Agenzien Temozolomid [TMZ] und Lomustin [CCNU]. Des Weiteren inaktivierten die Glukose-Konjugate die MGMT-Aktivität im Tumor eines Xenograft-Mausmodells und reduzierten das Tumorwachstum nach einer TMZ-Behandlung im gleichen Ausmass wie die Inhibitoren O6BG und O6BTG. Trotzdem war auch mit den Glukose-Konjugaten keine Steigerung der Zytostatika-Dosis im Mausmodell möglich. Die Untersuchungen der Aufnahme von O6BG-Glu und O6BTG-Glu wiederlegten eine Involvierung der Glukosetransporter. Der Einsatz von spezifischen Glukosetransporter-Inhibitoren und Kompetitions-Experimenten führte zu keiner Verminderung der MGMT-Hemmung oder Aufnahme vom radioaktiven H3-O6BTG-Glu in die Zelle. Dies legt nahe, dass die Glukose-Konjugate über einen unspezifischen Mechanismus (aktiv) in die Zellen gelangen. Der Grund für eine mögliche unselektive Aufnahme könnte im hydrophoben Alkyllinker, der für die Konjugation des Glukosemoleküls verwendet wurde, begründet sein. Dies führt zur Generierung von amphipathischen Konjugaten, die eine initiale Bindung an die Plasmamembran aufweisen und eine Aufnahme über den Flip-Flop-Mechanismus (transbilayer transport) wahrscheinlich machen. Die amphipathische Molekülstruktur der Glukose-Konjugate führte zu einer Partikelbildung in wässrigen Lösungen, die eine Reduktion der Menge an aktiven Monomeren von O6BG-Glu und O6BTG-Glu bewirken, die zur Hemmung von MGMT zur Verfügung stehen. Der zweite Teil der Arbeit befasste sich mit der Rolle von ABC-Transportern hinsichtlich einer Targeting-Strategie von MGMT-Hemmstoffen. Obwohl eine hohe Expression dieser ABC-Transporter in Tumoren zur Resistenzentwicklung gegenüber Zytostatika führt, wurde ihr Einfluss auf MGMT-Hemmstoffe oder einer MGMT-Targeting-Strategie niemals untersucht. In dieser Arbeit wurde zum ersten Mal ein aktiver Efflux von MGMT-Hemmstoffen durch ABC-Transporter nachgewiesen. Die Inhibition von ABC-Transportern bewirkte eine schnellere Inaktivierung von MGMT durch die Glukose-Konjugate. Des Weiteren zeigten Kompetitions-Experimente mit den MGMT-Hemmstoffen eine verminderte Efflux-Rate von Fluoreszenzfarbstoffen, die spezifisch von ABC-Transportern exportiert werden. ABC-Transporter reduzieren die wirksame Konzentration des Hemmstoffes in der Zelle und beeinträchtigen somit die Effektivität der MGMT-Inhibition. Eine simultane Hemmung der ABC-Transporter P-glycoprotein (P-gp), multi resistance protein 1 (MRP1) and breast cancer resistance protein (BCRP) erhöhte die Effektivität der MGMT-Hemmstoffe (O6BG, O6BTG, O6BG-Glu, O6BTG-Glu) und verstärkte auf diese Weise die TMZ-induzierte Toxizität in Tumorzelllinien. Die Involvierung von ABC-Transportern in der intrazellulären Speicherung von MGMT-Hemmstoffen ist wahrscheinlich die Ursache für die beobachteten Unterschiede in der Sensibilisierung verschiedener Tumorzelllinien gegenüber Zytostatika durch das Glukose-Konjugat O6BG-Glu. Eine Strategie, den Einfluss von ABC-Transportern zu reduzieren und zukünftliche MGMT-Targeting-Strategien effizienter umzusetzen, ist die Verwendung von O6BTG als Ausgangssubstanz. Die höhere Inhibitionsfähigkeit der Bromthiophenmoleküle vermindert die erforderliche intrazelluläre Konzentration für eine vollständige MGMT-Hemmung und reduziert auf diese Weise den Einfluss von ABC-Transportern.

Long-term cell-mediated protein release from calcium phosphate ceramics

Efficient delivery of growth factors from carrier biomaterials depends critically on the release kinetics of the proteins that constitute the carrier. Immobilizing growth factors to calcium phosphate ceramics has been attempted by direct adsorption and usually resulted in a rapid and passive release of the superficially adherent proteins. The insufficient retention of growth factors limited their bioavailability and their efficacy in the treatment of bone regeneration. In this study, a coprecipitation technique of proteins and calcium phosphate was employed to modify the delivery of proteins from biphasic calcium phosphate (BCP) ceramics. To this end, tritium-labeled bovine serum albumin ([(3)H]BSA) was utilized as a model protein to analyze the coprecipitation efficacy and the release kinetics of the protein from the carrier material. Conventional adsorption of [(3)H]BSA resulted in a rapid and passive release of the protein from BCP ceramics, whereas the coprecipitation technique effectively prevented the burst release of [(3)H]BSA. Further analysis of the in vitro kinetics demonstrated a sustained, cell-mediated release of coprecipitated [(3)H]BSA from BCP ceramics induced by resorbing osteoclasts. The coprecipitation technique described herein, achieved a physiologic-like protein release, by incorporating [(3)H]BSA into its respective carriers, rendering it a promising tool in growth factor delivery for bone healing.

Penetration of ASM 981 in canine skin: a comparative study

ASM 981 has been developed for topical treatment of inflammatory skin diseases. It specifically inhibits the production and release of pro-inflammatory cytokines. We measured the skin penetration of ASM 981 in canine skin and compared penetration in living and frozen skin. To make penetration of ASM 981 visible in dog skin, tritium labelled ASM 981 was applied to a living dog and to defrosted skin of the same dog. Using qualitative autoradiography the radioactive molecules were detected in the lumen of the hair follicles until the infundibulum, around the superficial parts of the hair follicles and into a depth of the dermis of 200 to 500 microm. Activity could not be found in deeper parts of the hair follicles, the dermis or in the sebaceous glands. Penetration of ASM 981 is low in canine skin and is only equally spread in the upper third of the dermis 24 hours after application. Penetration in frozen skin takes even longer than in living canine skin but shows the same distribution.

A gamma-glutamyl peptide isolated from onion (Allium cepa L.) by bioassay-guided fractionation inhibits resorption activity of osteoclasts

One gram of onion added to the food of rats inhibits significantly (p < 0.05) bone resorption as assessed by the urinary excretion of tritium released from bone of 9-week-old rats prelabeled with tritiated tetracycline from weeks 1 to 6. To isolate and identify the bone resorption inhibiting compound from onion, onion powder was extracted and the extract fractionated by column chromatography and medium-pressure liquid chromatography. A single active peak was finally obtained by semipreparative high-performance liquid chromatography. The biological activity of the various fractions was tested in vitro on the activity of osteoclasts to form resorption pits on a mineralized substrate. Medium, containing the various fractions or the pure compound, was added to osteoclasts of new-born rats settled on ivory slices. After 24 h of incubation, the tartrate-resistant acid phosphatase positive multinucleated cells, that is, osteoclasts, were counted. Subsequently, the number of resorption pits was determined. Activity was calculated as the ratio of resorption pits/osteoclasts and was compared to a negative control, that is, medium containing 10% fetal bovine serum only and to calcitonin (10(-12) M) as a positive control. Finally, a single peak inhibited osteoclast activity significantly (p < 0.05). The structure of this compound was elucidated with high-performance liquid chromatography-electrospray ionization-mass spectrometry, time-of-flight electrospray ionization mass spectrometry, and nuclear magnetic resonance spectroscopy. The single peak was identified as gamma-L-glutamyl-trans-S-1-propenyl-L-cysteine sulfoxide (GPCS). It has a molecular mass of 306 Da and inhibits dose-dependently the resorption activity of osteoclasts, the minimal effective dose being approximately 2 mM. As no other peak displayed inhibitory activity, it likely is responsible for the effect of onion on bone resorption.

Noble gas, CFC and other geochemical evidence for the age and origin of the Bath thermal waters, UK

The city of Bath is a World Heritage site and its thermal waters, the Roman Baths and new spa development rely on undisturbed flow of the springs (45 °C). The current investigations provide an improved understanding of the residence times and flow regime as basis for the source protection. Trace gas indicators including the noble gases (helium, neon, argon, krypton and xenon) and chlorofluorocarbons (CFCs), together with a more comprehensive examination of chemical and stable isotope tracers are used to characterise the sources of the thermal water and any modern components. It is shown conclusively by the use of 39Ar that the bulk of the thermal water has been in circulation within the Carboniferous Limestone for at least 1000 years. Other stable isotope and noble gas measurements confirm previous findings and strongly suggest recharge within the Holocene time period (i.e. the last 12 kyr). Measurements of dissolved 85Kr and chlorofluorocarbons constrain previous indications from tritium that a small proportion (<5%) of the thermal water originates from modern leakage into the spring pipe passing through Mesozoic valley fill underlying Bath. This introduces small amounts of O2 into the system, resulting in the Fe precipitation seen in the King’s Spring. Silica geothermometry indicates that the water is likely to have reached a maximum temperature of between 69–99 °C, indicating a most probable maximum circulation depth of ∼3 km, which is in line with recent geological models. The rise to the surface of the water is sufficiently indirect that a temperature loss of >20 °C is incurred. There is overwhelming evidence that the water has evolved within the Carboniferous Limestone formation, although the chemistry alone cannot pinpoint the geometry of the recharge area or circulation route. For a likely residence time of 1–12 kyr, volumetric calculations imply a large storage volume and circulation pathway if typical porosities of the limestone at depth are used, indicating that much of the Bath-Bristol basin must be involved in the water storage.

Identifiability of diffusion and sorption parameters from in situ diffusion experiments by using simultaneously tracer dilution and claystone data

In situ diffusion experiments are performed in geological formations at underground research laboratories to overcome the limitations of laboratory diffusion experiments and investigate scale effects. Tracer concentrations are monitored at the injection interval during the experiment (dilution data) and measured from host rock samples around the injection interval at the end of the experiment (overcoring data). Diffusion and sorption parameters are derived from the inverse numerical modeling of the measured tracer data. The identifiability and the uncertainties of tritium and Na-22(+) diffusion and sorption parameters are studied here by synthetic experiments having the same characteristics as the in situ diffusion and retention (DR) experiment performed on Opalinus Clay. Contrary to previous identifiability analyses of in situ diffusion experiments, which used either dilution or overcoring data at approximate locations, our analysis of the parameter identifiability relies simultaneously on dilution and overcoring data, accounts for the actual position of the overcoring samples in the claystone, uses realistic values of the standard deviation of the measurement errors, relies on model identification criteria to select the most appropriate hypothesis about the existence of a borehole disturbed zone and addresses the effect of errors in the location of the sampling profiles. The simultaneous use of dilution and overcoring data provides accurate parameter estimates in the presence of measurement errors, allows the identification of the right hypothesis about the borehole disturbed zone and diminishes other model uncertainties such as those caused by errors in the volume of the circulation system and the effective diffusion coefficient of the filter. The proper interpretation of the experiment requires the right hypothesis about the borehole disturbed zone. A wrong assumption leads to large estimation errors. The use of model identification criteria helps in the selection of the best model. Small errors in the depth of the overcoring samples lead to large parameter estimation errors. Therefore, attention should be paid to minimize the errors in positioning the depth of the samples. The results of the identifiability analysis do not depend on the particular realization of random numbers. (C) 2012 Elsevier B.V. All rights reserved.

Groundwater age distributions at a public drinking water supply well field derived from multiple age tracers (⁸⁵Kr, ³H/³He, and ³⁹Ar)

Groundwater age is a key aspect of production well vulnerability. Public drinking water supply wells typically have long screens and are expected to produce a mixture of groundwater ages. The groundwater age distributions of seven production wells of the Holten well field (Netherlands) were estimated from tritium-helium (3H/3He), krypton-85 (85Kr), and argon-39 (39Ar), using a new application of a discrete age distribution model and existing mathematical models, by minimizing the uncertainty-weighted squared differences of modeled and measured tracer concentrations. The observed tracer concentrations fitted well to a 4-bin discrete age distribution model or a dispersion model with a fraction of old groundwater. Our results show that more than 75 of the water pumped by four shallow production wells has a groundwater age of less than 20 years and these wells are very vulnerable to recent surface contamination. More than 50 of the water pumped by three deep production wells is older than 60 years. 3H/3He samples from short screened monitoring wells surrounding the well field constrained the age stratification in the aquifer. The discrepancy between the age stratification with depth and the groundwater age distribution of the production wells showed that the well field preferentially pumps from the shallow part of the aquifer. The discrete groundwater age distribution model appears to be a suitable approach in settings where the shape of the age distribution cannot be assumed to follow a simple mathematical model, such as a production well field where wells compete for capture area.

AN INVESTIGATION INTO THE MOLECULAR MECHANISMS OF ANDROGEN ACTION IN THE SERTOLI CELL

Steroid hormones regulate target cell function via quantitative and qualitative changes in RNA and protein synthesis. In the testis, androgens are known to play an important role in the regulation of spermatogenesis. The Sertoli cell (SC), whose function is thought to be supportive to the developing germ cell, has been implicated as an androgen target cell. Although cytoplasmic androgen receptors and chromatin acceptor sites for androgen-receptor complexes have been found in SC, effects on RNA synthesis have not previously been demonstrated. In this study, SC RNA synthetic activity was characterized and the effect of testosterone on SC nuclear transcriptional activity in vitro assessed. SC exhibited two fold increases in RNA and ribonucleotide pool concentrations during sexual maturation. These changes appeared to correlate with a previously observed increase in protein concentration per cell over an age span of 15-60 days. Following incubation with ('3)H-uridine, SC from older animals incorporated more label into RNA than SC from younger animals. Since the relative concentration of cytidine nucleotides was higher in SC from older rats, the age-related increase in tritium incorporation may reflect an associated increase in incorporation of ('3)H-CMP into RNA. Alternatively, the enhanced labeling may be the result of either a change in the base composition of the RNA resulting in a higher proportion of CMP and UMP in the RNA, or compartmentalization of the nucleotide pools. The physiologic consequences of these maturational alterations of nucleotide pools remains to be elucidated. RNA polymerase activities were characterized in intact nuclei obtained from cultured rat SC. (alpha)-Amanitin resistant RNA polymerase I+III activity was identical when measured in low or high ionic strength (0.05 M or 0.25 M ammonium sulfate (AS)) in the presence of MnCl(,2) or MgCl(,2), with a divalent cation optimum of 1.6 mM. RNA polymerase II was most active in 0.25 M AS and 1.6 mM MnCl(,2). The apparent Km of RNA polymerase II for UTP was 0.016 mM in 0.05 M AS and 0.037 mM in 0.25 M AS. The apparent Km values for total polymerase activity was 0.008 mM and 0.036 mM at low and high ionic strenghts, respectively. These data indicate that Sertoli cell RNA polymerase activities have catalytic properties characteristic of eukaryotic polymerase activities in general. In the presence of 21 (mu)M testosterone, RNA polymerase II activity increased two fold at 15 minutes, then declined but was still elevated over control values six hours after androgen addition. Polymerase I+III activity was not greatly affected by testosterone. The stimulation of polymerase II measured at 15 minutes was dose-dependent, with a maximum at 0.53 nM and no further stimulation up to 10('-5) M (ED(,50) = 0.25 nM testosterone), and was androgen specific. The results of preliminary RNA isolation and characterization experiments suggested that the synthesis of several species of RNA was enhanced by testosterone administration. These findings have great potential importance since they represent the first demonstration of a direct effect of androgens on the transcriptional process in the Sertoli cell. Furthermore, the results of these studies constitute further evidence that the Sertoli cell is a target for androgen action in the testis. ^