Measurement of cellular stimulation through monitoring pH changes by bead injection fluorescence microscopy

This work presents a method for extracellular and intracellular pH measurements in live cells based on a combination of the bead injection (BI) technique and fluorescence microscopy. For extracellular pH measurement, cells are grown on fluorescent beads, packed into a small column by a sequential injection instrument, and fluorescence intensity from the beads stained by the indicator is recorded by a fluorescence microscope. The method is applied to quantifying carbachol stimulation of Chinese hamster ovary (CHO) cells transfected with the m1 muscarinic receptor and is verified by a glucose depletion experiment. The results yield an EC(50) value of 1 muM for carbachol, which is in reasonable agreement with the literature value 3 muM determined by an existing potentiometric technique for measuring acid release. The intracellular measurement utilizes CHO M1 cells growing on non-fluorescent beads. For this method the cells rather than the beads are stained by incubating them in a solution of the fluorescent pH indicator BCECF. The cells are also stimulated with carbachol and the intracellular pH dependent fluorescence from the cells is recorded. The results show dependence between intracellular pH changes and carbachol concentration and yield an EC(50) value of 4 muM.     

An Earth-based equivalent low stretch apparatus for material flammability assessment in microgravity and extraterrestrial environments

The standard oxygen consumption (cone) calorimeter (described in ASTM E 1354 and NASA STD 6001 Test 2) is modified to provide a bench-scale test environment that simulates the low velocity buoyant or ventilation flow generated by or around a burning surface in a spacecraft or extraterrestrial gravity level. The equivalent low stretch apparatus (ELSA) uses an inverted cone geometry with the sample burning in a ceiling fire (stagnation flow) configuration. For a fixed radiant flux, ignition delay times for characterization material PMMA are shown to decrease by a factor of 3 at low stretch, demonstrating that ignition delay times determined from normal cone tests significantly underestimate the risk in microgravity. The critical heat flux for ignition is found to be lowered at low stretch as the convective cooling is reduced. At the limit of no stretch, any heat flux that exceeds the surface radiative loss at the surface ignition temperature is sufficient for ignition. Regression rates for PMMA increase with heat flux and stretch rate, but regression rates are much more sensitive to heat flux at the low stretch rates, where a modest increase in heat flux of 25 kW/m² increases the burning rates by an order of magnitude. The global equivalence ratio of these flames is very fuel rich, and the quantity of CO produced in this configuration is significantly higher than standard cone tests. These results demonstrate that the ELSA apparatus allows us to conduct normal gravity experiments that accurately and quantifiably evaluate a material’s flammability characteristics in the real-use environment of spacecraft or extraterrestrial gravitational acceleration. These results also demonstrate that current NASA STD 6001 Test 2 (standard cone) is not conservative since it evaluates a material’s flammability with a much higher inherent buoyant convective flow.     

Zur Bestimmung der Aufnahmef?higkeit der Ackererde an Wasser

Ohne Zusammenfassung     

Brouwer and Euclid

We explore the relationship between Brouwer’s intuitionistic mathematics and Euclidean geometry. Brouwer wrote a paper in 1949 called The contradictority of elementary geometry. In that paper, he showed that a certain classical consequence of the parallel postulate implies Markov’s principle, which he found intuitionistically unacceptable. But Euclid’s geometry, having served as a beacon of clear and correct reasoning for two millennia, is not so easily discarded.Brouwer started from a “theorem” that is not in Euclid, and requires Markov’s principle for its proof. That means that Brouwer’s paper did not address the question whether Euclid’s Elements really requires Markov’s principle. In this paper we show that there is a coherent theory of “non-Markovian Euclidean geometry”. We show in some detail that our theory is an adequate formal rendering of (at least) Euclid’s Book I, and suffices to define geometric arithmetic, thus refining the author’s previous investigations (which include Markov’s principle as an axiom).Philosophically, Brouwer’s proof that his version of the parallel postulate implies Markov’s principle could be read just as well as geometric evidence for the truth of Markov’s principle, if one thinks the geometrical “intersection theorem” with which Brouwer started is geometrically evident.     

Enantioselective Linchpin Catalysis by SOMO Catalysis: An Approach to the Asymmetric α‐Chlorination of Aldehydes and Terminal Epoxide Formation

Time for SOme MOre : For the first time SOMO (singly occupied molecular orbital) activation has been exploited to allow a new approach to the α‐chlorination of aldehydes. This transformation can be readily implemented as part of a linchpin catalysis approach to the enantioselective production of terminal epoxides.

     

Structural requirements for efficient photoinduced electron transfer (PET) quenching in 9-aminoalkylanthracenes

The structure-quenching relationship in 9-aminoalkylanthracenes is examined by the synthesis and fluorescence evaluation of five derivatives; it is observed that benzylic 2° and 3° amines lead to 95% quenching of anthracene fluorescence in water, while other amines afford substantially lower quenching efficiencies.     

Particle Filters with Nudging in Multiscale Chaotic Systems: With Application to the Lorenz ’96 Atmospheric Model

This paper presents reduced-order nonlinear filtering schemes based on a theoretical framework that combines stochastic dimensional reduction and nonlinear filtering. Here, dimensional reduction is achieved for estimating the slow-scale process in a multiscale environment by constructing a filter using stochastic averaging results. The nonlinear filter is approximated numerically using the ensemble Kalman filter and particle filter. The particle filter is further adapted to the complexities of inherently chaotic signals. In particle filters, an ensemble of particles is used to represent the distribution of the state of the hidden signal. The ensemble is updated using observation data to obtain the best representation of the conditional density of the true state variables given observations. Particle methods suffer from the “curse of dimensionality,” an issue of particle degeneracy within a sample, which increases exponentially with system dimension. Hence, particle filtering in high dimensions can benefit from some form of dimensional reduction. A control is superimposed on particle dynamics to drive particles to locations most representative of observations, in other words, to construct a better prior density. The control is determined by solving a classical stochastic optimization problem and implemented in the particle filter using importance sampling techniques.