Funktionelle Analyse der Plasmamembran-Ca 2+-ATPase 2 -PMCA2- in Modellen der Neurodegeneration

Calcium (Ca2+) ist ein ubiquitär vorkommendes Signalmolekül, das an der Regulation zahlreicher zellulärer Prozesse, von der Proliferation bis zum programmierten Zelltod, beteiligt ist. Daher müssen die intrazellulären Ca2+-Spiegel streng kontrolliert werden. Veränderungen der Ca2+-Homöostase während der altersassoziierten Neurodegeneration können dazu beitragen, dass Neuronen vulnerabler sind. So wurden erhöhte Ca2+-Konzentrationen in gealterten Neuronen, begleitet von einer erhöhten Vulnerabilität, beobachtet (Hajieva et al., 2009a). Weiterhin wird angenommen, dass der selektive Untergang von dopaminergen Neuronen bei der Parkinson Erkrankung auf eine erhöhte Ca2+-Last zurückzuführen sein könnte, da diese Neuronen einem ständigen Ca2+-Influx,rnaufgrund einer besonderen Isoform (CaV 1.3) spannungsgesteuerter Ca2+-Kanäle des L-Typs, ausgesetzt sind (Chan et al., 2007). Bislang wurden die molekularen Mechanismen, die einem Ca2+-Anstieg zu Grunde liegen und dessen Auswirkung jedoch nicht vollständig aufgeklärt und daher in der vorliegenden Arbeit untersucht. Um Veränderungen der Ca2+-Homöostase während der altersassoziiertenrnNeurodegeneration zu analysieren wurden primäre Mittelhirnzellen aus Rattenembryonen und SH-SY5Y-Neuroblastomazellen mit dem Neurotoxin 1-Methyl-4-Phenyl-Pyridin (MPP+), das bei der Etablierung von Modellen der Parkinson-Erkrankung breite Anwendung findet, behandelt. Veränderungen der intrazellulären Ca2+-Konzentration wurden mit einem auf dem grün fluoreszierenden Protein (GFP)-basierten Ca2+-Indikator,rn„Cameleon cpYC 3.6“ (Nagai et al., 2004), ermittelt. Dabei wurde in dieser Arbeit gezeigt, dass MPP+ die Abregulation der neuronenspezifischen ATP-abhängigen Ca2+-Pumpe der Plasmamembran (PMCA2) induziert, die mit der Ca2+-ATPase des endoplasmatischen Retikulums (SERCA) und dem Na+/Ca2+-Austauscher (NCX) das zelluläre Ca2+-Effluxsystem bildet, was zu einer erhöhten zytosolischen Ca2+-Konzentration führt. Die PMCA2-Abnahme wurde sowohl auf Transkriptionsebene als auch auf Proteinebene demonstriert, während keine signifikanten Veränderungen der SERCA- und NCX-Proteinmengen festgestellt wurden. Als Ursache der Reduktion der PMCA2-Expression wurde eine Abnahme des Transkriptionsfaktors Phospho-CREB ermittelt, dessen Phosphorylierungsstatus abhängig von der Proteinkinase A (PKA) war. Dieser Mechanismus wurde einerseits unter MPP+-Einfluss und andererseits vermittelt durch endogene molekulare Modulatoren gezeigt. Interessanterweise konnten die durch MPP+ induzierte PMCA2-Abregulation und der zytosolische Ca2+-Anstieg durch die Aktivierung der PKA verhindert werden. Parallel dazu wurde eine MPP+-abhängige verringerte mitochondriale Ca2+-Konzentration nachgewiesen, welche mit einer Abnahme des mitochondrialen Membranpotentials korrelierte. Darüber hinaus kam es als Folge der PMCA2-Abnahme zu einem verminderten neuronalen Überleben.rnVeränderungen der Ca2+-Homöostase wurden auch während der normalen Alterung inrnprimären Fibroblasten und bei Mäusen nachgewiesen. Dabei wurden verringerte PMCA und SERCA-Proteinmengen in gealterten Fibroblasten, einhergehend mit einem Anstieg der zytosolischen Ca2+-Konzentration demonstriert. Weiterhin wurden verringerte PMCA2-Proteinmengen im Mittelhirn von gealterten Mäusen (C57B/6) detektiert.rnDer zelluläre Ca2+-Efflux ist somit sowohl im Zuge der physiologischen Alterung als auch in einem altersbezogenen Krankheitsmodell beeinträchtigt, was das neuronale Überleben beeinflussen kann. In zukünftige Studien soll aufgeklärt werden, welche Auswirkungen einer PMCA2-Reduktion genau zu dem Verlust von Neuronen führen bzw. ob durch eine PMCA2-Überexpression neurodegenerative Prozesse verhindert werden können.

Frost heave testing of norwegian materials for road infrastructure

In areas of seasonal frost, frost susceptibility composed by frost heaving during the winter and thaw softening during the spring is one of the most dangerous phenomenon for transportation, road and railway infrastructure. Therefore, the need for frost protection layer becomes imperative. The purpose of frost protection layer is to prevent frost from penetrating down through the pavement and into the sub-soils. Frost susceptible soils under the road can be cause damages on the roads or other structures due to frost heave or reduced capacity characteristics thaw period. "Frost heave" is the term given to the upwards displacement of the ground surface caused by the formation of ice within soils or aggregates (Rempel et al., 2004). Nowadays in Scandinavia the most common material used in frost protection layer in the pavement structure of roads and in the ballast of the railway tracks are coarse-grain crushed rocks aggregates. Based on the capillary rise, the mechanics of frost heave phenomenon is based on the interaction between aggregates and water, as suggested by Konrad and Lemieux in 2005 that said that the fraction of material below the 0.063 mm sieve for coarse-grained soils must be controlled so as to reduce the sensitivity to frost heave. The study conducted in this thesis project is divided in two parts: - the analysis of the coarse grained aggregates used in frost protection layer in Norway; - the analysis of the frost heave phenomenon in the laboratory under known boundary conditions, through the use of the most widely used method, the frost heave test, in” closed system” (without access of water).

Studio di flussi granulari secchi riprodotti in laboratorio a media scala: Elaborazione di un modello applicabile a casi naturali.

Questa tesi di laurea si riferisce allo studio cinematico e morfologico di flussi granulari composti di materiale vulcanico naturale tramite la strumentazione sperimentale GRANFLOW-sim presso il Dipartimento di Scienze Geologiche dell’Università Autonoma di San Luis Potosì in Messico. E’ stato adottato un approccio sperimentale finalizzato a riprodurre le condizioni fisico-meccaniche di flussi granulari secchi in piccola scala riconducibili a differenti ambienti deposizionali naturali e artificiali. La strumentazione è composta da una canaletta di alimentazione ad inclinazione fissa di 40° ed un piano di deposizione a inclinazioni variabile da 0° a 20° al fine di simulare diverse condizioni deposizionali. Specifici sensori in posizione fissa sullo strumento hanno consentito di raccogliere dati cinematici del flusso e morfologici del deposito corrispondente, rielaborati in profili di velocità e modelli morfologici digitali per ognuno degli esperimenti effettuati. E’ stata osservata una tripartizione del fronte del flusso granulare caratterizzata da i) clasti balistici isolati, ii) un fronte disperso costituito da clasti saltellanti che precede l’arrivo del iii) fronte compatto del flusso costituito dalla parte principale del materiale trasportato. La normalizzazione dei dati cinematici e morfologici ha reso possibile l’elaborazione di relazioni empiriche per lunghezza e larghezza del deposito e velocità massima del flusso in funzione di variabili condizioni di slope-ratio (rapporto tra l’inclinazione del piano di deposizione e della canaletta) e ΔH (differenza di quota tra l’area sorgente del flusso ed il fronte compatto del deposito). Queste relazioni sono state confrontate con i valori ottenuti da Zanchetta et al. (2004) per i flussi granulari naturali nell’area di Sarno, unico caso di studio reale confrontabile con le condizioni sperimentali adottate in questa tesi. Tale confronto ha mostrato una buona corrispondenza tra dati sperimentali e situazione reale nei casi in cui l’effetto di canalizzazione dei flussi granulari era trascurabile. Ciò evidenzia la possibilità di calibrare la strumentazione GRANFLOW-sim al fine di incorporare la valutazione dell’effetto di canalizzazione, ampliando il raggio di utilizzo delle relazioni empiriche ad un numero maggiore di casi di studio di flussi granulari naturali.

Bringing results to fruition through publication: An analysis of a peer-reviewed, open access and context-focused journal's editorial practice

When it comes to helping to shape sustainable development, research is most useful when it bridges the science–implementation/management gap and when it brings development specialists and researchers into a dialogue (Hurni et al. 2004); can a peer-reviewed journal contribute to this aim? In the classical system for validation and dissemination of scientific knowledge, journals focus on knowledge exchange within the academic community and do not specifically address a ‘life-world audience’. Within a North-South context, another knowledge divide is added: the peer review process excludes a large proportion of scientists from the South from participating in the production of scientific knowledge (Karlsson et al. 2007). Mountain Research and Development (MRD) is a journal whose mission is based on an editorial strategy to build the bridge between research and development and ensure that authors from the global South have access to knowledge production, ultimately with a view to supporting sustainable development in mountains. In doing so, MRD faces a number of challenges that we would like to discuss with the td-net community, after having presented our experience and strategy as editors of this journal. MRD was launched in 1981 by mountain researchers who wanted mountains to be included in the 1992 Rio process. In the late 1990s, MRD realized that the journal needed to go beyond addressing only the scientific community. It therefore launched a new section addressing a broader audience in 2000, with the aim of disseminating insights into, and recommendations for, the implementation of sustainable development in mountains. In 2006, we conducted a survey among MRD’s authors, reviewers, and readers (Wymann et al. 2007): respondents confirmed that MRD had succeeded in bridging the gap between research and development. But we realized that MRD could become an even more efficient tool for sustainability if development knowledge were validated: in 2009, we began submitting ‘development’ papers (‘transformation knowledge’) to external peer review of a kind different from the scientific-only peer review (for ‘systems knowledge’). At the same time, the journal became open access in order to increase the permeability between science and society, and ensure greater access for readers and authors in the South. We are currently rethinking our review process for development papers, with a view to creating more space for communication between science and society, and enhancing the co-production of knowledge (Roux 2008). Hopefully, these efforts will also contribute to the urgent debate on the ‘publication culture’ needed in transdisciplinary research (Kueffer et al. 2007).

A Faster Circular Binary Segmentation Algorithm for the Analysis of Array CGH Data

Motivation: Array CGH technologies enable the simultaneous measurement of DNA copy number for thousands of sites on a genome. We developed the circular binary segmentation (CBS) algorithm to divide the genome into regions of equal copy number (Olshen {\it et~al}, 2004). The algorithm tests for change-points using a maximal $t$-statistic with a permutation reference distribution to obtain the corresponding $p$-value. The number of computations required for the maximal test statistic is $O(N^2),$ where $N$ is the number of markers. This makes the full permutation approach computationally prohibitive for the newer arrays that contain tens of thousands markers and highlights the need for a faster. algorithm. Results: We present a hybrid approach to obtain the $p$-value of the test statistic in linear time. We also introduce a rule for stopping early when there is strong evidence for the presence of a change. We show through simulations that the hybrid approach provides a substantial gain in speed with only a negligible loss in accuracy and that the stopping rule further increases speed. We also present the analysis of array CGH data from a breast cancer cell line to show the impact of the new approaches on the analysis of real data. Availability: An R (R Development Core Team, 2006) version of the CBS algorithm has been implemented in the ``DNAcopy'' package of the Bioconductor project (Gentleman {\it et~al}, 2004). The proposed hybrid method for the $p$-value is available in version 1.2.1 or higher and the stopping rule for declaring a change early is available in version 1.5.1 or higher.

1,25-(OH)2-16ene-23yne-D3 reduces secondary hyperparathyroidism in uremic rats with little calcemic effect

To compare the effects of vitamin D analogs versus calcitriol on serum levels of Ca, P and parathyroid hormone (PTH). A compound better than calcitriol should increase the Ca x P product less than calcitriol for an equivalent decrease in PTH levels.

Regulation of the Telecommunications in Switzerland: A Network Approach to Assess the Regulatory Agencies' Independence

The liberalization process of the Swiss telecommunications sector follows a logic of ‘autonomous adaptation’ to the regulations of the European Union (EU). Switzerland, which is not a Member State of the EU, voluntarily adapts to the European policy without being for- mally required to do so (Sciarini et al., 2004). This process went hand in hand with the partial privatization of the legal statute and assets of the former monopolist and with the re-regulation of the liberalized telecommunications sector.

North Pacific Carbon Cycle Response to Climate Variability on Seasonal to Decadal Timescales

Climate variability drives significant changes in the physical state of the North Pacific, and there may be important impacts of this variability on the upper ocean carbon balance across the basin. We address this issue by considering the response of seven biogeochemical ocean models to climate variability in the North Pacific. The models' upper ocean pCO(2) and air-sea CO(2) flux respond similarly to climate variability on seasonal to decadal timescales. Modeled seasonal cycles of pCO(2) and its temperature- and non-temperature-driven components at three contrasting oceanographic sites capture the basic features found in observations (Takahashi et al., 2002, 2006; Keeling et al., 2004; Brix et al., 2004). However, particularly in the Western Subarctic Gyre, the models have difficulty representing the temporal structure of the total pCO(2) seasonal cycle because it results from the difference of these two large and opposing components. In all but one model, the air-sea CO(2) flux interannual variability (1 sigma) in the North Pacific is smaller ( ranges across models from 0.03 to 0.11 PgC/yr) than in the Tropical Pacific ( ranges across models from 0.08 to 0.19 PgC/yr), and the time series of the first or second EOF of the air-sea CO(2) flux has a significant correlation with the Pacific Decadal Oscillation (PDO). Though air-sea CO(2) flux anomalies are correlated with the PDO, their magnitudes are small ( up to +/- 0.025 PgC/yr ( 1 sigma)). Flux anomalies are damped because anomalies in the key drivers of pCO(2) ( temperature, dissolved inorganic carbon (DIC), and alkalinity) are all of similar magnitude and have strongly opposing effects that damp total pCO(2) anomalies.

Risk factors for developing complications while awaiting a cholecystectomy

Background. Laparoscopic Cholecystectomy is the gold standard for patients who are diagnosed with biliary colic (NIH, 1993). It has been demonstrated that individuals who wait a longer time between diagnosis and treatment are at increased risk of having complications (Rutledge et al., 2000; Contini et al., 2004; Eldar et al., 1999). County hospitals, such as Ben Taub General Hospital (BTGH), have a particularly high population of uninsured patients and consequently long surgical wait periods due to limited resources. This study evaluates patients the risk factors involved in their progression to complications from gallstones in a county hospital environment. ^ Methods. A case-control study using medical records was performed on all patients who underwent a cholecystectomy for gallstone disease at BTGH during the year of 2005 (n=414). The risk factors included in the study are obesity, gender, age, race, diabetes, and amount of time from diagnosis to surgery. Multivariate analysis and logistical regression were used to assess factors that potentially lead to the development of complications. ^ Results. There were a total of 414 patients at BTGH who underwent a cholecystectomy for gallstone disease during 2005. The majority of patients were female, 84.3% (n=349) and Hispanic, 79.7% (n=330). The median wait time from diagnosis to surgery was 1.43 weeks (range: 0-184.71). The majority of patients presented with complications 72.5% (n=112). The two factors that impacted development of complications in our study population were Hispanic race (OR=1.81; CI 1.02, 3.23; p=0.04) and time from diagnosis to surgery (OR=0.98; CI 0.97, 0.99; p<0.01). Obesity, gender, age, and diabetes were not predictive of development of complications. ^ Conclusions. An individual's socioeconomic status potentially influences all aspects of their health and subsequent health care. The patient population of BTGH is largely uninsured and therefore less likely to seek care at an early stage in their disease process. In order to decrease the rate of complications, there needs to be a system that increases patient access to primary care clinics. Until the problem of access to care is solved, those who are uninsured will likely suffer more severe complications and society will bear the cost. ^

Plasma homocysteine levels and Alzheimer's Disease: A systematic review

Objective. To systematically review studies published in English on the relationship between plasma total homocysteine (Hcy) levels and the clinical and/or postmortem diagnosis of Alzheimer's disease (AD) in subjects who are over 60 years old.^ Method. Medline, PubMed, PsycINFO and Academic Search Premier, were searched by using the keywords "homocysteine", "Alzheimer disease" and "dementia", and "cognitive disorders". In addition, relevant articles in PubMed using the "related articles" link and by cross-referencing were identified. The study design, study setting and study population, sample size, the diagnostic criteria of the National Institute of Neurological and Communicative Disorders and Stroke (NINCDS) and the Alzheimer's Disease and Related Disorders Association (ADRDA), and description of how Hcy levels were measured or defined had to have been clearly stated. Empirical investigations reporting quantitative data on the epidemiology of the relationship between plasma total Hcy (exposure factor) and AD (outcome) were included in the systematic review.^ Results. A total of 7 studies, which included a total of 2,989 subjects, out of 388 potential articles met the inclusion criteria: four case control and three cohort studies were identified. All 7 studies had association statistics, such as the odds ratio (OR), the relative rates (RR), and the hazard ratio (HR) of AD, examined using multivariate and logistic regression analyses. Three case - comparison studies: Clarke et al. (1998) (OR: 4.5, 95% CI.: 2.2 - 9.2); McIlroy et al. (2002) (OR: 2.9, 95% CI.: 1.00–8.1); Quadri et al. (2004) (OR: 3.7, 95% CI.: 1.1 - 13.1), and two cohort studies: Seshadri et al. (2002) (RR: 1.8, 95% CI.: 1.3 - 2.5); Ravaglia et al. (2005) (HR: 2.1, 95% CI.: 1.7 - 3.8) found a significant association between serum total Hcy and AD. One case-comparison study, Miller et al. (2002) (OR: 2.2, 95% C.I.: 0.3 -16), and one cohort study, Luchsinger et al. (2004) (HR: 1.4, 95% C.I.: 0.7 - 2.3) failed to reject H0.^ Conclusions. The purpose of this review is to provide a thorough analysis of studies that examined the relationship between Hcy levels and AD. Five studies showed a positive statistically significant association between elevated total Hcy values and AD but the association was not statistically significant in two studies. Further research is needed in order to establish evidence of the strong, consistent association between serum total Hcy and AD as well as the presence of the appropriate temporal relationship. To answer these questions, it is important to conduct more prospective studies that examine the occurrence of AD in individuals with and without elevated Hcy values at baseline. In addition, the international standardization of measurements and cut-off points for plasma Hcy levels across laboratories is a critical issue to be addressed for the conduct of future studies on the topic.^

Mid-Pliocene palynomorph record of the North Atlantic

The mid-Pliocene was an episode of prolonged global warmth and strong North Atlantic thermohaline circulation, interrupted briefly at circa 3.30 Ma by a global cooling event corresponding to marine isotope stage (MIS) M2. Paleoceanographic changes in the eastern North Atlantic have been reconstructed between circa 3.35 and 3.24 Ma at Deep Sea Drilling Project Site 610 and Integrated Ocean Drilling Program Site 1308. Mg/Ca ratios and d18O from Globigerina bulloides are used to reconstruct the temperature and relative salinity of surface waters, and dinoflagellate cyst assemblages are used to assess variability in the North Atlantic Current (NAC). Our sea surface temperature data indicate warm waters at both sites before and after MIS M2 but a cooling of ~2-3°C during MIS M2. A dinoflagellate cyst assemblage overturn marked by a decline in Operculodinium centrocarpum reflects a southward shift or slowdown of the NAC between circa 3.330 and 3.283 Ma, reducing northward heat transport 23-35 ka before the global ice volume maximum of MIS M2. This will have established conditions that ultimately allowed the Greenland ice sheet to expand, leading to the global cooling event at MIS M2. Comparison with an ice-rafted debris record excludes fresh water input via icebergs in the northeast Atlantic as a cause of NAC decline. The mechanism causing the temporary disruption of the NAC may be related to a brief reopening of the Panamanian Gateway at about this time.

Stable oxygen isotope record and Sr/Ca ratios of corals from IODP Holes M0009D and M0023A

We present monthly resolved records of strontium/calcium (Sr/Ca) and oxygen isotope (d18O) ratios from well-preserved fossil corals drilled during the Integrated Ocean Drilling Program (IODP) Expedition 310 'Tahiti Sea Level' and reconstruct sea surface conditions in the central tropical South Pacific Ocean during two time windows of the last deglaciation. The two Tahiti corals examined here are uranium/thorium (U/Th)-dated at 12.4 and 14.2 ka, which correspond to the Younger Dryas (YD) cold reversal and the Bølling-Allerød (B-A) warming of the Northern Hemisphere, respectively. The coral Sr/Ca records indicate that annual average sea surface temperature (SST) was 2.6-3.1 °C lower at 12.4 ka and 1.0-1.6 °C lower at 14.2 ka relative to the present, with no significant changes in the amplitude of the seasonal SST cycle. These cooler conditions were accompanied by seawater d18O (d18Osw) values higher by ~0.8 per mill and ~0.6 per mill relative to the present at 12.4 and 14.2 ka, respectively, implying more saline conditions in the surface waters. Along with previously published coral Sr/Ca records from the island [Cohen and Hart (2004), Deglacial sea surface temperatures of the western tropical Pacific: A new look at old coral. Paleoceanography 19, PA4031, doi:10.1029/2004PA001084], our new Tahiti coral records suggest that a shift toward lower SST by ~1.5 °C occurred from 13.1 to 12.4 ka, which was probably associated with a shift toward higher d18Osw by ~0.2 per mill. Along with a previously published coral Sr/Ca record from Vanuatu [Corrège et al. (2004), Interdecadal variation in the extent of South Pacific tropical waters during the Younger Dyras event. Nature 428, 927-929], the Tahiti coral records provide new evidence for a pronounced cooling of the western to central tropical South Pacific during the Northern Hemisphere YD event.

Chronology for the EDML ice core from Dronning Maud Land, Antarctica, over the last 150 000 years

A chronology called EDML1 has been developed for the EPICA ice core from Dronning Maud Land (EDML). EDML1 is closely interlinked with EDC3, the new chronology for the EPICA ice core from Dome-C (EDC) through a stratigraphic match between EDML and EDC that consists of 322 volcanic match points over the last 128 ka. The EDC3 chronology comprises a glaciological model at EDC, which is constrained and later selectively tuned using primary dating information from EDC as well as from EDML, the latter being transferred using the tight stratigraphic link between the two cores. Finally, EDML1 was built by exporting EDC3 to EDML. For ages younger than 41 ka BP the new synchronized time scale EDML1/EDC3 is based on dated volcanic events and on a match to the Greenlandic ice core chronology GICC05 via 10Be and methane. The internal consistency between EDML1 and EDC3 is estimated to be typically ~6 years and always less than 450 years over the last 128 ka (always less than 130 years over the last 60 ka), which reflects an unprecedented synchrony of time scales. EDML1 ends at 150 ka BP (2417 m depth) because the match between EDML and EDC becomes ambiguous further down. This hints at a complex ice flow history for the deepest 350 m of the EDML ice core.