Liquidus Tracking: Controlled Rate Vitrification for the Cryopreservation of Larger Volumes and Tissues

BACKGROUND: Vitrification of cells or tissue at controlled cooling rates suitable for larger volumes, and with reduced cryoprotectant toxicity. OBJECTIVE: To set out the current understanding of the LiquidusTracking (LT) vitrification technique, and to discuss the challenges and benefits of translating the method into laboratory protocols more generally applicable to meet requirements of large volume and 3-D cryo-banking in the era of regenerative medicine. METHODS: By adding small amounts of cryoprotectants at each step and subsequently cooling the sample just above its freezing point before further increasing CPA concentration, cryoprotectant toxicity is minimized. RESULT: CPA toxicity can be reduced by lowering the temperature. Different manual approaches to LT were evaluated and further improved. CONCLUSIONS: Manual liquidus tracking is complicated and exhibits potential high variability. Nevertheless, this approach offers the possibility of testing several conditions simultaneously and could be used to pre-test conditions prior to automatic LT development

It’s all about information? The Following Behaviour of Professors and PhD Students on Twitter

In this paper we investigate the role of the academic status in the following behaviour of computer scientists on Twitter. Based on a uses and gratifications perspective, we focus on the activity of a Twitter account and the reciprocity of following relationships. We propose that the account activity addresses the users' information motive only, whereas the user's academic status relates to both the information motive and community development (as in peer networking or career planning). Variables were extracted from Twitter user data. We applied a biographical approach to correctly identify the academic status (professor versus PhD student). We calculated a $2\times 2$ MANOVA on the influence of the activity of the account and the academic status (on different groups of followers) to differentiate the influence of the information motive versus the motive for community development. Results suggest that for computer scientists Twitter is mainly an information network. However, we found significant effects in the sense of career planning, that is, the accounts of professors had even in the case of low activity a relatively high number of researcher followers -- both PhD followers as well as professor followers. Additionally, there was also some weak evidence for community development gratifications in the sense of peer-networking of professors. Overall, we conclude that the academic use of Twitter is not only about information, but also about career planning and networking

Analyse der räumlichen Variabilität von Makroporenstrukturen in Lössböden anhand geostatistischer Auswertungen von 3D-Röntgen-CT Bildern

Eigenschaften von Porennetzwerken (z.B. Konnektivität und Tortuosität) und deren Funktionen (z.B. Transport von Sauerstoff und Wasser) sind von großer Bedeutung für das Pflanzenwachstum. Die unterschiedliche und sich überlagernde Genese von Porenräumen in Böden, z.B. durch Wurzeln und Regenwürmer entstandene Bioporen oder Quellungs- und Schrumpfungsrisse, führen zu äußerst komplexen und räumlich heterogenen dreidimensionalen Porenraumstrukturen, die mit einfachen Parametern wie Porositäten oder Porengrößenverteilungen nur unzureichend beschrieben sind. Vor dem Hintergrund, dass moderne bildgebende Technologien wie die Röntgen-Computertomographie zunehmend zur Analyse und Quantifizierung von Porenraumeigenschaften und ihrer Beziehung zu Bodenfunktionen zum Einsatz kommen, ergeben sich vielfältige Möglichkeiten der bildanalytischen Auswertung der generierten Datensätze. Dabei ist eine genauere Beschreibung der räumlichen Heterogenität und die getrennte Auswertung von unterschiedlich entstandenen Porenräumen (abiotisch versus biotisch) von Interesse, um deren relativen Beitrag zu porenskaligen Prozessen (Austauschvorgänge an Poren/Matrix-Grenzflächen, Transportfunktionen, etc.) besser quantifizieren zu können.  In dieser Studie wurden Bodenmonolithe (Ø 20 cm, 70 cm Höhe) aus einem Feldversuch in der Nähe von Bonn (NRW, Deutschland) verwendet. Die Bodenmonolithe wurden aus dem Unterboden (45 - 105 cm) eines Versuches mit unterschiedlicher Wurzelausprägung durch Vorfrüchte entnommen und mit einem industriellen Röntgen-CT gescannt. Mittels 3D-Bildanalyse und geostatistischer Methoden wurden quantitative und qualitative Parameter des Porenraums untersucht. Dabei konnten Unterschiede in der Bodenstruktur, in Abhängigkeit der Wurzelsysteme der Vorfrüchte, festgestellt werden. Insbesondere konnte die Häufigkeit, Größenverteilung und Raumverteilung (randomisiert, gruppiert, etc.) der Porenstrukturen festgestellt werden. Weiter konnten   Anisotropieeigenschaften und Hauptorientierungsrichtungen der Makroporen ermittelt werden. Anhand dieser morphologischen Information war es möglich die Makroporen in Bioporen (hauptsächlich Wurzelpfade und Regenwurmgänge) und abiotische Strukturporen (hauptsächlich durch quellen und schrumpfen) zu unterteilen. Aufgrund der unterschiedlichen Wurzelausprägung in den untersuchten Proben, zeigten sich Unterschiede in diesen Porenkategorien. Inwieweit sich diese Porenkategorien  funktionell unterscheiden und welchen Einfluss sie jeweils auf den Wasser-, Gas- und Nährstoffhaushalt von Böden haben soll diskutiert werden

Small-scale swirl events in the quiet Sun chromosphere

Recent progress in instrumentation enables solar observations with high resolution simultaneously in the spatial, temporal, and spectral domains. We use such high-resolution observations to study small-scale structures and dynamics in the chromosphere of the quiet Sun. We analyze time series of spectral scans through the Ca II 854.2nm spectral line obtained with the CRISP instrument at the Swedish 1-m Solar Telescope. The targets are quiet Sun regions inside coronal holes close to disc-centre. The line core maps exhibit relatively few fibrils compared to what is normally observed in quiet Sun regions outside coronal holes. The time series show a chaotic and dynamic scene that includes spatially confined "swirl" events. These events feature dark and bright rotating patches, which can consist of arcs, spiral arms, rings or ring fragments. The width of the fragments typically appears to be on the order of only 0.2", which is close to the effective spatial resolution. They exhibit Doppler shifts of -2 to -4 km/s but sometimes up to -7 km/s, indicating fast upflows. The diameter of a swirl is usually of the order of 2". At the location of these swirls, the line wing and wide-band maps show close groups of photospheric bright points that move with respect to each other. A likely explanation is that the relative motion of the bright points twists the associated magnetic field in the chromosphere above. Plasma or propagating waves may then spiral upwards guided by the magnetic flux structure, thereby producing the observed intensity signature of Doppler-shifted ring fragments.Comment: 4 pages, 3 figures, A&A Letter, accepted (final version

Trimerisation of carbon suboxide at a di-titanium centre to form a pyrone ring system

The reaction of the syn-bimetallic bis(pentalene)dititanium complex Ti2(μ:η5,η5-Pn†)2 (Pn† = C8H4(1,4-SiiPr3)2) 1 with carbon suboxide (O[double bond, length as m-dash]C[double bond, length as m-dash]C[double bond, length as m-dash]C[double bond, length as m-dash]O, C3O2) results in trimerisation of the latter and formation of the structurally characterised complex [{Ti2(μ:η5,η5-Pn†)2}{μ-C9O6}]. The trimeric bridging C9O6 unit in the latter contains a 4-pyrone core, a key feature of both the hexamer and octamer of carbon suboxide which are formed in the body from trace amounts of C3O2 and are, for example, potent inhibitors of Na+/K+-ATP-ase. The mechanism of this reaction has been studied in detail by DFT computational studies, which also suggest that the reaction proceeds via the initial formation of a mono-adduct of 1 with C3O2. Indeed, the carefully controlled reaction of 1 with C3O2 affords [Ti2(μ:η5,η5-Pn†)2 (η2-C3O2)], as the first structurally authenticated complex of carbon suboxide

Liquidus Tracking: Large scale preservation of encapsulated 3-D cell cultures using a vitrification machine

Currently, cryo-banking of multicellular structures such as organoids, especially in large volumes at clinical scale >1 L, remains elusive for reasons such as insufficient dehydration and cryoprotectant additive (CPA(1)) penetration, slow cooling and warming rates and devitrification processes. Here we introduce the concept of Liquidus Tracking (LT) using a semi-automated process for liquid volumes of up to 450 ml including 130 ml of alginate encapsulated liver cells (AELC) that archived controlled and reversible vitrification with minimized toxicity. First a CPA solution with optimal properties for LT was developed by employing different small scale test systems. Combining sugars such as glucose and raffinose with Me2SO improved post-exposure (at +0.5 °C) viabilities from 6% ±3.6 for Me2SO alone up to 58% ±6.1 and 65% ±14.2 respectively (p < 0.01). Other permeating CPAs (e.g. ethylene glycol, propylene glycol, methanol) were investigated as partial replacements for Me2SO. A mixture of Me2SO, ethylene glycol and glucose (ratio 4:2:1- termed LTdeg) supported glass-forming tendencies with appropriate low viscosities and toxicities required for LT. When running the full LT process, using Me2SO alone, no viable cells were recovered; using LTdeg, viable recoveries were improved to 40% ±8 (p<0.001%). Further refinements of improved mixing technique further improved recovery after LT. Recoveries of specific liver cell functions such as synthesis of albumin and alpha-fetoprotein (AFP) were retained in post thaw cultures. In summary: By developing a low-toxicity CPA solution of low viscosity (LTdeg) suitable for LT and by improving the stirring system, post-warming viability of AELC of up to 90% and a AFP secretion of 89% were reached. Results show that it may be possible to develop LT as a suitable cryogenic preservation process for different cell therapy products at large scale

3D photospheric velocity field of a Supergranular cell

We investigate the plasma flow properties inside a Supergranular (SG) cell, in particular its interaction with small scale magnetic field structures. The SG cell has been identified using the magnetic network (CaII wing brightness) as proxy, applying the Two-Level Structure Tracking (TST) to high spatial, spectral and temporal resolution observations obtained by IBIS. The full 3D velocity vector field for the SG has been reconstructed at two different photospheric heights. In order to strengthen our findings, we also computed the mean radial flow of the SG by means of cork tracing. We also studied the behaviour of the horizontal and Line of Sight plasma flow cospatial with cluster of bright CaII structures of magnetic origin to better understand the interaction between photospheric convection and small scale magnetic features. The SG cell we investigated seems to be organized with an almost radial flow from its centre to the border. The large scale divergence structure is probably created by a compact region of constant up-flow close to the cell centre. On the edge of the SG, isolated regions of strong convergent flow are nearby or cospatial with extended clusters of bright CaII wing features forming the knots of the magnetic network.Comment: 7 pages, submitted to A&A, referee's comments include

How Digital Are the Digital Humanities? An Analysis of Two Scholarly Blogging Platforms

In this paper we compare two academic networking platforms, HASTAC and Hypotheses, to show the distinct ways in which they serve specific communities in the Digital Humanities (DH) in different national and disciplinary contexts. After providing background information on both platforms, we apply co-word analysis and topic modeling to show thematic similarities and differences between the two sites, focusing particularly on how they frame DH as a new paradigm in humanities research. We encounter a much higher ratio of posts using humanities-related terms compared to their digital counterparts, suggesting a one-way dependency of digital humanities-related terms on the corresponding unprefixed labels. The results also show that the terms digital archive, digital literacy, and digital pedagogy are relatively independent from the respective unprefixed terms, and that digital publishing, digital libraries, and digital media show considerable cross-pollination between the specialization and the general noun. The topic modeling reproduces these findings and reveals further differences between the two platforms. Our findings also indicate local differences in how the emerging field of DH is conceptualized and show dynamic topical shifts inside these respective contexts

Ein neuer Ansatz für die Trockenseparierung von Mikroaggregaten mit unterschiedlicher Textur zur Messung der mechanischen Belastbarkeit und 3D-Porenstruktur

Die Bodenstruktur als Ausdruck der räumlichen Anordnung mineralischer und organischer Bodenbestandteile ist eine zentrale Charakteristik des Bodens. Sie steuert viele wichtige biologische, physikalische und geochemische Prozesse, wie die Rolle des Bodens als Kohlenstoffspeicher oder die Ausbildung bzw. Verteilung von Habitaten für Mikroorganismen. Die Bodenstruktur, deren einfachste Einheit die Aggregate bilden, befindet sich als labile Bodeneigenschaft in einem Zustand ständiger Veränderung. Die Eigenschaften der Aggregate werden durch viele Einflussfaktoren wie Textur, Alter, Quellung und Schrumpfung, sowie die biologische Aktivität gesteuert. Eines der Hauptprobleme bei der Untersuchung der Eigenschaften von Mikroaggregaten im Boden ist deren Separierung. Viele Separierungs-Methoden üben Spannungszustände aus, die die realen Bedingungen im Boden nur sehr bedingt abbilden. So werden z. B. bei Nasssiebungsverfahren hydraulische Spannungen erzeugt, die unter natürlichen Bedingungen nicht auftreten. Hierin liegt ein Risiko, dass Artefakte in den gewonnenen Aggregatfraktionen entstehen (z. B. durch Reaggregierung bei anschließender Trocknung) und die weitere Analyse von Eigenschaften dieser Aggregatfraktionen, bzw. deren Interpretation beeinflussen. Übergeordnetes Ziel unserer Untersuchungen ist die Erforschung der Genese von Mikroaggregaten und deren (Poren‑)Eigenschaften in Abhängigkeit von Texturunterschieden, sowie des Zusammenhangs von mikroskaligen Deformationsprozessen auf die Entwicklung der Bodenstruktur. Hierfür haben wir mit einem Verfahren der Trockenseparierung in drei Aggregatgrößen-Unterklassen (250-53, 53-20 und &lt;20 µm) eine zuverlässige Methode zur Isolierung einzelner Mikroaggregate entwickelt, welche die Struktur der gewonnenen Aggregate selbst nicht beeinflusst. In einem nächsten Schritt wird die mechanische Belastbarkeit von Mikroaggregaten aus einer Toposequenz (mit unterschiedlichen Tongehalten) an einem Lastrahmen hochauflösend getestet, um die Hypothese zu überprüfen, dass die Stabilität mit abnehmender struktureller Entropie (d. h. zunehmendem Grad an Strukturierung) zunimmt. Des Weiteren wird die Geometrie des Porennetzwerkes der Mikroaggregate mit unterschiedlichen Tongehalten mittels hochauflösender Computertomographie untersucht, um diese später mit gemessenen Gas- und Wasserflüssen in Verbindung bringen zu können

The GREGOR Fabry-P\'erot Interferometer

The GREGOR Fabry-P\'erot Interferometer (GFPI) is one of three first-light instruments of the German 1.5-meter GREGOR solar telescope at the Observatorio del Teide, Tenerife, Spain. The GFPI uses two tunable etalons in collimated mounting. Thanks to its large-format, high-cadence CCD detectors with sophisticated computer hard- and software it is capable of scanning spectral lines with a cadence that is sufficient to capture the dynamic evolution of the solar atmosphere. The field-of-view (FOV) of 50" x 38" is well suited for quiet Sun and sunspot observations. However, in the vector spectropolarimetric mode the FOV reduces to 25" x 38". The spectral coverage in the spectroscopic mode extends from 530-860 nm with a theoretical spectral resolution R of about 250,000, whereas in the vector spectropolarimetric mode the wavelength range is at present limited to 580-660 nm. The combination of fast narrow-band imaging and post-factum image restoration has the potential for discovery science concerning the dynamic Sun and its magnetic field at spatial scales down to about 50 km on the solar surface.Comment: 14 pages, 17 figures, 4 tables; pre-print of AN 333, p.880-893, 2012 (AN special issue to GREGOR