Regional phospholipid analysis of porcine lens membranes by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

The applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) to the qualitative and quantitative analysis of most mammalian phospholipid (PL) classes was demonstrated in a crude extract of porcine lens membranes. When 2,5-dihydroxybenzoic acid (DHB) was used as the matrix, positive-ion spectra allowed the accurate quantification of phosphatidylcholines (PCs) and sphingomyelins (SMs). Other PLs such as phosphatidylethanolamines (PEs), phosphatidylethanolamine plasmalogens (PEps), phosphatidylethanolamine ethers (PEes) and phosphatidylserines (PSs), could also be detected, but their lower ionization efficiency led to negative errors in their quantification. Despite this limitation, it was possible to determine relative changes among PLs extracted from cortical and nuclear regions. Negative-ion spectra were acquired with the use of p-nitroaniline (PNA) as the matrix. Because neither PCs nor SMs produce negative ions, other PL classes can be analyzed selectively. The absolute quantification of the various PL classes detectable in negative-ion spectra was also affected by differences in ionization efficiencies. However, the trends in compositional changes between cortical and nuclear-fiber PLs were in agreement with those obtained by (31)P NMR spectroscopy. MALDI-TOFMS also offers the possibility of studying variations in the acyl-chain distribution of the various species comprising each PL class. For porcine lenses, PCs, PEs and phosphatidylinositols (PIs) exhibited the greatest depletions in going from cortical to nuclear membranes. Among their individual species, those with two or more sites of unsaturation suffered the most significant reduction.     

Inhibition of peroxyoxalate chemiluminescence by intercalation of fluorescent acceptors between DNA bases

We have examined the ability of different fluorescent DNA dyes to become chemically excited by the peroxyoxalate chemiluminescent reaction. The intercalating dyes ethidium bromide and propidium iodide, and the bis-intercalating dyes ethidium homodimer-1, benzoxazolium-4-pyridinium dimer-1 and benzoxazolium-4-quinolinium dimer-1, exhibit an intense chemiluminescence when they are excited by the bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H2O2 reaction in the absence of DNA. However, the chemiluminescence of these dyes is very low when they are bound to double-stranded DNA (dsDNA). In contrast, the minor groove-binding dye Hoechst 33258 excited by the TCPO-H2O2 reaction shows approximately the same chemiluminescence intensity when it is free in solution or complexed with dsDNA. Structural alterations or partial dissociation of dsDNA-bis-intercalating dye complexes produced by the addition of acetone, NaCl, MgCl2 or the cationic surfactant cetyltrimethylammonium bromide increases the chemiluminescence intensity. A moderate chemiluminescence intensity is observed when bis-intercalating dyes are complexed with single-stranded DNA. Our results indicate that the energy from the intermediates produced in the peroxyoxalate chemiluminescent reaction cannot be efficiently transferred to fluorescent dyes complexed with DNA; chemiexcitation is almost completely inhibited when dyes are buried in the dsDNA structure by intercalation between the base pairs.     

Micellar-stabilized room-temperature phosphorimetric determination of the fungicide thiabendazole in canned pineapple samples

A method for the determination of the fungicide thiabendazole (TBZ) by micellar-stabilized room-temperature phosphorescence is described. It does not require any separation step and allows the direct determination of the fungicide in canned pineapple samples. The effect of various experimental conditions is discussed in detail. The analytical curve of thiabendazole gives a linear dynamic range of 23.8–500.0 ng mL^–1 and a detection limit of 23.8 ng mL^–1. Recoveries of 103.9 and 89.2% for syrup and canned pineapple pulp, respectively, were obtained for 250 ng mL^–1 thiabendazole. Received: 30 April 1997 / Revised: 18 July 1997 / Accepted: 23 July 1997     

Modeling diamagnetic and magnetooptic properties of organic compounds with the TOSS-MODE approach

The topological substructural molecular design (TOSS-MODE) approach is used to describe the diamagnetic susceptibility of organic compounds. Two data sets composed of 233 aliphatic and 85 aromatic compounds were studied for which good linear correlations were found. The contributions of many different structural fragments and atomic groups were computed by the current approach. The predictive ability of the models developed was tested by using external prediction sets of compounds of different classes than those used in training. A quantitative model based on the current approach was developed to compute the diamagnetic susceptibility exaltation of aromatic compounds, which is exemplified by the study of polycyclic aromatic hydrocarbons. The rotatory power of organic compounds in a magnetic field was also described by the TOSS-MODE approach. Good linear correlations were obtained for this property in aliphatic and aromatic compounds. The predictive abilities of the models found were tested by external prediction sets for which good correlations between calculated and experimental values are found.     

New hierarchy for the Liouville equation,irreversibility and Fokker‐Planck‐like structures

The issue of irreversibility is revisited for a closed system formed by N classical non‐relativistic particles inside a volume Ω, interacting through two‐body potentials, for large N and Ω. The classical phase‐space distribution function f, multiplied by suitable Hermite polynomials and integrated over all momenta, yields new moments. The Liouville equation and the initial distribution f_in imply a new non‐equilibrium linear infinite hierarchy for the moments. That hierarchy differs from the BBGKY one for distribution functions and displays some suggestive Fokker‐Planck‐like structures. A physically motivated ansatz for f_in (which introduces statistical assumptions), used by previous authors, is chosen. All moments of order n ≥ n₀ are expressed in terms of those of order n₀ — 1 and of f_in. The properties of the Fokker‐Planck‐like structures (hermiticity, non‐negative eigenvalues) allow for implementing a natural long‐time approximation in the hierarchy, so as to introduce relaxation to equilibrium and irreversibility, consistently with the hydrodynamical balance equations. Further (more restrictive) assumptions and approximations lead to new irreversible models, generalizing non‐trivially the Fokker‐Planck equation. They are described through a truncated hierarchy of linear equations for moments of order n ≤ n₀ — 1 (n₀ being finite). The connections with Brownian particle dynamics and Fluid Dynamics are analyzed, for consistency.     

Protein Backbone N Relaxation Rates as a Tool for the Diagnosis of Structure Quality

In the work reported herein we define a structure validation factor that depends on protein backbone ¹⁵N relaxation rates. This is an alternative method to the previously defined quality factors derived from anisotropic chemical shifts or residual dipolar couplings. We have used the structure dependence of ¹⁵N relaxation rates of anisotropically tumbling proteins to calculate this structure diagnosis factor and have used it to demonstrate the improvement of protein structures refined with residual dipolar couplings.     

Measurement of Signal‐to‐Noise Ratio In Graphene‐based Passive Microelectrode Arrays

This work aims to investigate the influence of various electrode materials on the signal‐to‐noise ratio (SNR) of passive microelectrode arrays (MEAs) intended for use in neural interfaces. Noise reduction substantially improves the performance of systems which electrically interface with extracellular solutions. The MEAs are fabricated using gold, indium tin oxide (ITO), inkjet printed (IJP) graphene, and chemical vapor deposited (CVD) graphene. 3D‐printed Nylon reservoirs are adhered to glass substrates with identical MEA patterns and filled with neuronal cell culture media. To precisely control the electrode area and minimize the parasitic coupling of metal interconnects and solution, SU‐8 photoresist is patterned to expose only the area of the electrode to solution and cap the remainder of the sample. Voltage signals with varying amplitude and frequencies are applied to the solution using glass micropipettes, and the response is measured on an oscilloscope from a microprobe placed on the contact pad external to the reservoir. The time domain response signal is transformed into a frequency spectrum, and SNR is calculated. As the magnitude or the frequency of the input signal gets larger, a significantly increased signal‐to‐noise ratio was observed in CVD graphene MEAs compared to others. This result indicates that 2‐dimensional nanomaterials such as graphene can provide better signal integrity and potentially lead to improved performance in hybrid neural interface systems.     

Yttrium Geothermometry Applied to Garnets from Different Metamorphic Grades Analysed by EPMA and µ-PIXE Techniques

We present a multi-analytical and multi-instrumental approach to a petrologic study of garnets from metapelites characterized by different metamorphic grades. The comparison between major, minor and trace element (Y) distribution in garnets crystallized at different temperatures was performed by electron microprobe (EDS and WDS) and the Proton Induced X-ray Emission microprobe (μ-PIXE). Two garnet samples were selected from metapelite rocks at different metamorphic grades from the tectonic unit of Stilo, Calabrian–Peloritanian Arc, Southern Italy. Quantitative spot analysis profiles and compositional X-ray maps of major elements and yttrium are reported. The major element zoning of garnets is mostly characterized by a smoothed and gradual concentric variation of all elements from core to rim. The Y concentration reveals a marked zoning with its distribution decreasing from about 7000 ppm to about 100 ppm from core to rim, respectively. The precise determination of the Y content by μ-PIXE technique allowed us to better define the rock thermal history, by applying the Y geothermometer. In particular, a common starting temperature of about 450 °C was calculated for the beginning of the garnet growth from all over the unit, but different temperatures in the range from 520 °C to 670 °C were estimated for the peak event in garnets from different areas, in agreement with the P–T values given by the main rock mineral assemblages, reflecting a different metamorphic grade.