Monitoring cellular responses upon fatty acid exposure by Fourier transform infrared spectroscopy and Raman spectroscopy

We investigated the applicability of FTIR-spectroscopy as a high throughput screening method for detection of biochemical changes in intact liver cells in bulk upon fatty acid exposure. HepG2 cells adapted to serum free (HepG2-SF) growth were exposed to four different fatty acids, three octadecenoic acids, differing in cis/trans-configuration or double bond position (oleic acid, elaidic acid and vaccenic acid) as well as palmitic acid in three days. High throughput FTIR spectroscopic measurements on dried films of intact cells showed spectra with high signal-to-noise ratio and great reproducibility. When applying principal component analysis (PCA) a clear discrimination between fatty acid exposures was observed. Higher levels of triacylglycerides were accumulated in cells exposed to elaidic acid than when exposed to the other fatty acids; the least accumulation appeared to be in cells exposed to palmitic acid. An increased absorption at ~966 cm(-1) corresponding to trans-double bond was detected upon elaidic acid exposure but not upon vaccenic acid exposure. Instead, upon vaccenic acid exposure two new absorption bands were observed at 981 and 946 cm(-1) due to the presence of double bond conjugation. Raman spectroscopy on single cells, with and without treatment by vaccenic acid, confirmed the presence of conjugation. By fatty acid composition analysis, the conjugation was further specified to be conjugated linoleic acid (CLA) isomers. Thus, instead of being preserved as a monounsaturated fatty acid, vaccenic acid was converted into CLA in HepG2 cells. The results demonstrate the applicability of high-throughput FTIR spectroscopy as an explorative method in in vitro systems from which biologically relevant hypotheses can be generated and further investigated.     

Experimental evaluation of X‐ray optics applied for microanalysis

The performance of x‐ray capillary lenses has been evaluated. The tests were carried out using an x‐ray tube set‐up. A single glass capillary with tapered inner channel, a monolithic glass polycapillary, and an in‐house manufactured single metallic capillary with parabolic inner channel were characterized in terms of gain, spatial resolution, and element detection limits. The spatial resolution of a confocal set‐up utilizing a monolithic glass polycapillary and a polycapillary conical collimator has also been measured. The highest gain of about 2500 was observed for the glass polycapillary. The maximum gain achieved with the single glass capillary was equal to about 25, and the gain of the metallic capillary was only slightly greater than 1. For the glass capillary and polycapillary lenses, significant filtering of the higher‐energy photons (energy > 8 keV) was observed. The lowest relative detection limits were obtained with an ordinary cylindrical collimator and the polycapillary lens. Similar absolute detection limits were achieved with the use of the polycapillary and single capillary lenses. A relation between the ratios of the detection limits of elements achieved with different x‐ray lenses and the lens parameters (spatial resolution and gain) has been proposed and was verified experimentally. The monolithic polycapillary lens was found to be an optimum focusing device for an x‐ray tube‐based scanning spectrometer. This type of x‐ray lens can be coupled with a polycapillary conical collimator or a polycapillary half‐lens to make a confocal x‐ray microscope capable of depth profiling with a spatial resolution equal to about 30 micrometers. Copyright © 2008 John Wiley & Sons, Ltd.     

Meissner effect in superconducting microtraps

We report on the realization and characterization of a magnetic microtrap for ultracold atoms near a straight superconducting Nb wire with circular cross section. The trapped atoms are used to probe the magnetic field outside the superconducting wire. The Meissner effect shortens the distance between the trap and the wire, reduces the radial magnetic-field gradients, and lowers the trap depth. Measurements of the trap position reveal a complete exclusion of the magnetic field from the superconducting wire for temperatures lower than 6 K. As the temperature is further increased, the magnetic field partially penetrates the superconducting wire; hence the microtrap position is shifted towards the position expected for a normal-conducting wire.     

Disorder and decision cost in spatial networks

We introduce the concept of decision cost of a spatial graph, which measures the disorder of a given network taking into account not only the connections between nodes but their position in a two-dimensional map. The influence of the network size is evaluated and we show that normalization of the decision cost allows us to compare the degree of disorder of networks of different sizes. Under this framework, we measure the disorder of the connections between airports of two different countries and obtain some conclusions about which of them is more disordered. The introduced concepts (decision cost and disorder of spatial networks) can easily be extended to Euclidean networks of higher dimensions, and also to networks whose nodes have a certain fitness property (i.e., one-dimensional).     

The Role of Entropy in Construct Specification Equations (CSE) to Improve the Validity of Memory Tests: Extension to Word Lists

Metrological methods for word learning list tests can be developed with an information theoretical approach extending earlier simple syntax studies. A classic Brillouin entropy expression is applied to the analysis of the Rey’s Auditory Verbal Learning Test RAVLT (immediate recall), where more ordered tasks—with less entropy—are easier to perform. The findings from three case studies are described, including 225 assessments of the NeuroMET2 cohort of persons spanning a cognitive spectrum from healthy older adults to patients with dementia. In the first study, ordinality in the raw scores is compensated for, and item and person attributes are separated with the Rasch model. In the second, the RAVLT IR task difficulty, including serial position effects (SPE), particularly Primacy and Recency, is adequately explained (Pearson’s correlation R=0.80) with construct specification equations (CSE). The third study suggests multidimensionality is introduced by SPE, as revealed through goodness-of-fit statistics of the Rasch analyses. Loading factors common to two kinds of principal component analyses (PCA) for CSE formulation and goodness-of-fit logistic regressions are identified. More consistent ways of defining and analysing memory task difficulties, including SPE, can maintain the unique metrological properties of the Rasch model and improve the estimates and understanding of a person’s memory abilities on the path towards better-targeted and more fit-for-purpose diagnostics.     

Theoretical study of the mechanism of the Wittig reaction: Ab initio and MNDO-PM3 treatment of the reaction of unstabilized,semistabilized, and stabilized ylides with acetaldehyde

In this study, we describe the results of ab initio (HF and MP2) and MNDO-PM3 calculations on the model reactions of unstabilized (Me₃P=CH–CH₃), semi-stabilized (Me₃P=CH–C≡CH), and stabilized (Me₃P=CH–C≡N) ylides with acetaldehyde to form their respective Z and E olefins and trimethylphosphine oxide. These reactions occur in three stages: oxaphosphetane formation, oxaphosphetane pseudorotation, and oxaphosphetane decomposition. The calculated barriers for these processes vary considerably depending on the level of theory employed (ab initio vs. MNDO-PM3 or HF vs. MP2 at the ab initio level). However, self-consistent geometries of reactants, intermediates, transition states and products are obtained at all levels. Oxaphosphetane formation is best described as very asynchronous cycloaddition (borderline two-step mechanisms). The geometries of the transition states are near planar with respect to P, C, C, and O atoms. Analysis of the bond indices of these reactions shows that the C–C bonds are between 44% (unstabilized case) and 60% (stabilized case) formed, whereas the corresponding P–O bonds have not been formed to any significant degree. Oxaphosphetane decomposition can be described as a very asynchronous retrocycloaddition where P–C bond breakage runs ahead of C–O bond breakage. These results are compared with experimental findings for the Wittig reaction, and its relevance to the overall mechanism of the olefination is discussed. © 1997 John Wiley & Sons, Inc. Heteroatom Chem 8: 557–569, 1997