High-Level Expression of Various Apolipoprotein (a) Isoforms by "Transferrinfection". The Role of Kringle IV Sequences in the Extracellular Association with Low-Density Lipoprotein

Characterization of the assembly of lipoprotein(a) [Lp(a)] is of fundamental importance to understanding the biosynthesis and metabolism of this atherogenic lipoprotein. Since no established cell lines exist that express Lp(a) or apolipoprotein(a) [apo(a)], a "transferrinfection" system for apo(a) was developed utilizing adenovirus receptor- and transferrin receptor-mediated DNA uptake into cells. Using this method, different apo(a) cDNA constructions of variable length, due to the presence of 3, 5, 7, 9, 15, or 18 internal kringle IV sequences, were expressed in cos-7 cells or CHO cells. All constructions contained kringle IV-36, which includes the only unpaired cysteine residue (Cys-4057) in apo(a). r-Apo(a) was synthesized as a precursor and secreted as mature apolipoprotein into the medium. When medium containing r-apo(a) with 9, 15, or 18 kringle IV repeats was mixed with normal human plasma LDL, stable complexes formed that had a bouyant density typical of Lp(a). Association was substantially decreased if Cys-4057 on r-apo(a) was replaced by Arg by site-directed mutagenesis or if Cys-4057 was chemically modified. Lack of association was also observed with r-apo(a) containing only 3, 5, or 7 kringle IV repeats without "unique kringle IV sequences", although Cys-4057 was present in all of these constructions. Synthesis and secretion of r-apo(a) was not dependent on its sialic acid content. r-Apo(a) was expressed even more efficiently in sialylation-defective CHO cells than in wild-type CHO cells. In transfected CHO cells defective in the addition of N-acetylglucosamine, apo(a) secretion was found to be decreased by 50%. Extracellular association with LDL was not affected by the carbohydrate moiety of r-apo(a), indicating a protein-protein interaction between r-apo(a) and apoB. These results show that, besides kringle IV-36, other kringle IV sequences are necessary for the extracellular association of r-apo(a) with LDL. Changes in the carbohydrate moiety of apo(a), however, do not affect complex formation

Studies on the heterogeneity of human serum Lp lipoproteins and on the occurrence of double Lp lipoprotein variants

Lp lipoproteins have been prepared by a mild method from the serum of a large number of individuals. Approximately 25% of the individuals tested showed the presence of a double Lp peak in analytical ultracentrifuge diagrams. These double peaks were designated Lp(a)-1 and Lp(a)-2 to distinguish them from the single Lp(a) peak. The mean viscosity-corrected sedimentation coefficient, S 1.004, 20 C and density of the single Lp(a) peak were 15.8±1.8 s ( n =32) and 1.076±0.01 g/ml, of the Lp(a)-1 peak were 13.5±1.1 s ( n =14) and 1.064±0.007 g/ml, and of the Lp(a)-2 peak were 16.8±1.7 s ( n =14) and 1.074±0.009 g/ml. Absorption tests using a double and single Lp preparation showed that both Lp peaks in the double variants possess Lp(a) specificity. Evidence is lacking as yet for individual specificities for either Lp(a)-1 or Lp(a)-2. Interand intra-individual heterogeneity among Lp lipoproteins is discussed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/44186/1/10528_2004_Article_BF00485737.pd

Spectral Statistics in Chaotic Systems with Two Identical Connected Cells

Chaotic systems that decompose into two cells connected only by a narrow channel exhibit characteristic deviations of their quantum spectral statistics from the canonical random-matrix ensembles. The equilibration between the cells introduces an additional classical time scale that is manifest also in the spectral form factor. If the two cells are related by a spatial symmetry, the spectrum shows doublets, reflected in the form factor as a positive peak around the Heisenberg time. We combine a semiclassical analysis with an independent random-matrix approach to the doublet splittings to obtain the form factor on all time (energy) scales. Its only free parameter is the characteristic time of exchange between the cells in units of the Heisenberg time.Comment: 37 pages, 15 figures, changed content, additional autho

Comparative Microbiome and Metabolome Analyses of the Marine Tunicate Ciona intestinalis from Native and Invaded Habitats

Massive fouling by the invasive ascidian Ciona intestinalis in Prince Edward Island (PEI, Canada) has been causing devastating losses to the local blue mussel farms. In order to gain first insights into so far unexplored factors that may contribute to the invasiveness of C. intestinalis in PEI, we undertook comparative microbiome and metabolome studies on specific tissues from C. intestinalis populations collected in invaded (PEI) and native regions (Helgoland and Kiel, Germany). Microbial community analyses and untargeted metabolomics revealed clear location- and tissue-specific patterns showing that biogeography and the sampled tissue shape the microbiome and metabolome of C. intestinalis. Moreover, we observed higher microbial and chemical diversity in C. intestinalis from PEI than in the native populations. Bacterial OTUs specific to C. intestinalis from PEI included Cyanobacteria (e.g., Leptolyngbya sp.) and Rhodobacteraceae (e.g., Roseobacter sp.), while populations from native sampling sites showed higher abundances of e.g., Firmicutes (Helgoland) and Epsilonproteobacteria (Kiel). Altogether 121 abundant metabolites were putatively annotated in the global ascidian metabolome, of which 18 were only detected in the invasive PEI population (e.g., polyketides and terpenoids), while six (e.g., sphingolipids) or none were exclusive to the native specimens from Helgoland and Kiel, respectively. Some identified bacteria and metabolites reportedly possess bioactive properties (e.g., antifouling and antibiotic) that may contribute to the overall fitness of C. intestinalis. Hence, this first study provides a basis for future studies on factors underlying the global invasiveness of Ciona species

Mobilität von CeO2-Nanomaterialien in Böden

Der zunehmende Einsatz von Cerdioxid-Nanomaterialien (CeO2 NM) führt zu ihrer Freisetzung in die aquatische und terrestrische Umwelt. Aufgrund der ungeklärten human- und umwelttoxischen Wirkung von CeO2 NM muss deren Verbleib in der Umwelt untersucht werden. Es wurden zwei verschiedene CeO2 NM (OECD-Standard CeO2 NM-212, Europium-gedopte CeO2 NM (5 % Eu)) zu elf luftgetrockneten Ackerböden pulverförmig zugegeben und mit künstlichem Regenwasser dispergiert (Batchversuche), um Nachweismethoden für Bodenfestphase und Bodenlösung zu entwickeln. Darüber hinaus sollen diese Batchversuche erste Informationen über die Mobilität der NM in Böden liefern. Zusätzlich wurden Säulenversuche mit einer schluffigen Parabraunerde durchgeführt, um die Wiederfindung und mögliche Remobilisierung des Eu-gedopten CeO2 NM in Böden bzw. Bodenlösungen mit hohen Ce-Hintergrundgehalten zu untersuchen. Nach der pulverförmigen Applikation des gedopten NM wurde die Parabraunerde jeweils für 3 und 92 Tage inkubiert, der Wassergehalt wurde wöchentlich auf 55 % der maximalen Wasserhaltekapazität eingestellt. Anschließend erfolgte die Elution unter gesättigten Bedingungen mit künstlichem Regenwasser unter Austausch von maximal zwölf Porenvolumina. Die Bodenlösungen der Batchversuche und die Säuleneluate wurden nach der Fraktionierung (Filtration, Zentrifugation und Ultrazentrifugation) mit HNO3 und H2O2 aufgeschlossen. Die Messung der Ce- und Eu-Gehalte erfolgte mit ICP-OES und ICP-MS. Durch das Eu-Doping wird weder das Kristallgitter noch die Größe der Primärpartikel verändert. Die Bestimmungsgrenzen der applizierten NM in Böden und Bodenlösungen werden im Vergleich zum ungedopten Material, insbesondere bei hohen Ce-Hintergrundgehalten, deutlich erniedrigt. Dadurch können umweltrelevante Konzentrationen des NM getestet werden. Batchversuche sind für die Untersuchung der Mobilität von NM stark limitiert, dennoch deutet die variierende Freisetzung der NM aus den verschiedenen Bodenproben darauf hin, dass die unterschiedlichen Bodencharakteristika die NM Mobilität steuern. Die CeO2 NM Freisetzung aus den Säulenversuchen lag auf einem sehr niedrigen Niveau, war nach der Langzeitinkubation jedoch minimal erhöht. Aus den Ergebnissen lässt sich folgern, dass Böden wahrscheinlich eine deutliche Senke für die getesteten NM darstellen

Bose-Einstein condensates in a double well: mean-field chaos and multi-particle entanglement

A recent publication [Phys. Rev. Lett. 100, 140408 (2008)] shows that there is a relation between mean-field chaos and multi-particle entanglement for BECs in a periodically shaken double well. 'Schrodinger-cat' like mesoscopic superpositions in phase-space occur for conditions for which the system displays mean-field chaos. In the present manuscript, more general highly-entangled states are investigated. Mean-field chaos accelerates the emergence of multi-particle entanglement; the boundaries of stable regions are particularly suited for entanglement generation.Comment: 5 Pages, 5 jpg-figures, to be published in the proceedings of the LPHYS0

Effect of dissipation and measurement on a tunneling system

We consider a parametrically driven Kerr medium in which the pumping may be sinusoidally varied. It has been previously found that this system exhibits coherent tunneling between two fixed points which can be either enhanced or suppressed by altering the driving frequency and strength. We numerically investigate the dynamics when damping is included. This is done both by solving a master equation and using the quantum-trajectory method. In the latter case it is also possible to model the result of a continuous heterodyne measurement of the cavity output. The dissipation destroys the coherences which give rise to the tunneling, causing the sinusoidal oscillation of the mean to give way to a stochastic jumping between the fixed points, manifested as a random telegraph signal. In the quantum-trajectory picture we show that the coherences responsible for tunneling are an exponentially decreasing function of the signal-to-noise ratio for heterodyne measurements. However, evidence of both the bare tunneling rate and the driving modified tunneling rate are still apparent in the random telegraph signal

Dynamical Tunneling in Mixed Systems

We study quantum-mechanical tunneling in mixed dynamical systems between symmetry-related phase space tori separated by a chaotic layer. Considering e.g. the annular billiard we decompose tunneling-related energy splittings and shifts into sums over paths in phase space. We show that tunneling transport is dominated by chaos-assisted paths that tunnel into and out of the chaotic layer via the ``beach'' regions sandwiched between the regular islands and the chaotic sea. Level splittings are shown to fluctuate on two scales as functions of energy or an external parameter: they display a dense sequence of peaks due to resonances with states supported by the chaotic sea, overlaid on top of slow modulations arising from resonances with states supported by the ``beaches''. We obtain analytic expressions which enable us to assess the relative importance of tunneling amplitudes into the chaotic sea vs. its internal transport properties. Finally, we average over the statistics of the chaotic region, and derive the asymptotic tail of the splitting distribution function under rather general assumptions concerning the fluctuation properties of chaotic states.Comment: 28 pages, Latex, 16 EPS figure

Tunneling of quantum rotobreathers

We analyze the quantum properties of a system consisting of two nonlinearly coupled pendula. This non-integrable system exhibits two different symmetries: a permutational symmetry (permutation of the pendula) and another one related to the reversal of the total momentum of the system. Each of these symmetries is responsible for the existence of two kinds of quasi-degenerated states. At sufficiently high energy, pairs of symmetry-related states glue together to form quadruplets. We show that, starting from the anti-continuous limit, particular quadruplets allow us to construct quantum states whose properties are very similar to those of classical rotobreathers. By diagonalizing numerically the quantum Hamiltonian, we investigate their properties and show that such states are able to store the main part of the total energy on one of the pendula. Contrary to the classical situation, the coupling between pendula necessarily introduces a periodic exchange of energy between them with a frequency which is proportional to the energy splitting between quasi-degenerated states related to the permutation symmetry. This splitting may remain very small as the coupling strength increases and is a decreasing function of the pair energy. The energy may be therefore stored in one pendulum during a time period very long as compared to the inverse of the internal rotobreather frequency.Comment: 20 pages, 11 figures, REVTeX4 styl