Circular Dichroism Spectral Similarity Plots to Extend Validation and Correction to All Measured Wavelengths

Interlaboratory comparisons of circular dichroism (CD) spectra are useful for developing confidence in the measurements associated with optically active molecules. These measurements also help define the higher-order (secondary and tertiary) structure of biopolymers. Unfortunately, the extent of the validity of these measurements has been unclear. In this work, a method is described to extend CD validation over the entire observed wavelength range using what will be called spectral similarity plots. The method involves plotting, wavelength by wavelength, all measured spectral intensities of a sample at one concentration against the intensity values of the same material at a different concentration or pathlength. These spectral similarity plots validate the instrument in terms of spectral shape and whether the shape is shifted in intensity and/or in wavelength. This comparison tests the linearity of instrument’s signal, the balance of its left and right polarizations, its wavelengths, and its spectral intensity scales. When the process is applied to materials with accepted and archived intensity values, the method can be linked to older single-wavelength and double-wavelength calibration techniques. Further, spectral similarity testing of CD spectra from samples with different concentrations run in different labs suggests that improved interlaboratory validation of CD data is possible. Since a database of archival CD measurements is available online, spectral similarity comparisons could possibly provide the ability to compare linearity, polarization balance, wavelength, and spectral intensity between all current CD instruments. If the preliminary results published here prove robust and transferable, then comparisons of full-wavelength range spectra to archived data using spectral similarity plots should become part of the standard process to validate and calibrate the performance of CD instruments.

PATH-45. APOLLO: RAMAN-BASED PATHOLOGY OF MALIGNANT GLIOMA

BACKGROUND DNA methylation is an essential component for integrative diagnosis in glioma. Methylation subtype prediction of gliomas is currently done via sample extraction of high-quality of reasonable amount of DNA (~1ug), methylome profiling, followed by probe identification, curation and subsequent analysis via different random forest classifiers. However, the DNA methylation classification is not always available for all the samples. METHODS Raman Spectroscopy performed of the regions of interest using 1mm2 FFPE tissue spots from 45 patient samples with LGm1 to LGm6 methylation subtypes. Spectral information was then used to train a convolutional neural network (CNN) and develop a prediction algorithm. 70 % of dataset - model training while the remaining 30% for validation. Supervised wrapper methods and random forests were used to identify the top 109 most discriminatory Raman frequencies out of 1738. RESULTS We identified the most discriminatory features from these analyses and demonstrated that these frequencies show differential spectral intensities for these frequencies depending upon the glioma subtypes across the larger areas of the tissue. We compared the results of the Ward linkage clustering with the separation induced by the “frequency criterion”, an empirical observation that Raman spectra of tumor spots are characterized by intensities higher than 5000 on some of the frequencies from 1463 to 1473. For each of the 45 samples we ran Ward linkage clustering with a variable number of clusters (from 2 to 7), with the majority cluster corresponding to tumor spots and the others corresponding to (various types of) non-tumor spots. We found that the majority cluster matches very well the tumor spots characterized by the frequency criterion, The average accuracy over all samples was 90:3%, the average precision was 99:6% and the average recall was 90:2%. For most samples, two clusters were sufficient to distinguish between tumor and non-tumor spots with accuracy.

A Ratiometric Optical Dual Sensor for the Simultaneous Detection of Oxygen and Carbon Dioxide

Simultaneous detection of carbon dioxide (CO2) and oxygen (O2) has attracted considerable interest since CO2 and O2 play key roles in various industrial and domestic applications. In this study, a new approach based on a fluorescence ratiometric referencing method was reported to develop an optical dual sensor where platinum (II) meso-tetrakis(pentafluorophenyl)porphyrin (PtTFPP) complex used as the O2-sensitive dye, CdSe/ZnS quantum dots (QDs) combined with phenol red used as the CO2-sensitive dye, and CdSe/ZnS QDs used as the reference dye for the simultaneous detection of O2 and CO2. All the dyes were immobilized in a gas-permeable matrix poly (isobutyl methacrylate) (PolyIBM) and subjected to excitation using a 380 nm LED. The as-obtained distinct fluorescence spectral intensities were alternately exposed to analyte gases to observe changes in the fluorescence intensity. In the presence of O2, the fluorescence intensity of the Pt (II) complex was considerably quenched, while in the presence of CO2, the fluorescence intensity of QDs was increased. The corresponding ratiometric sensitivities of the optical dual sensor for O2 and CO2 were approximately 13 and 144, respectively. In addition, the response and recovery for O2 and CO2 were calculated to be 10 s/35 s and 20 s/60 s, respectively. Thus, a ratiometric optical dual gas sensor for the simultaneous detection of O2 and CO2 was successfully developed. Effects of spurious fluctuations in the intensity of external and excitation sources were suppressed by the ratiometric sensing approach.

EXPRESS: Laser-Induced Breakdown Spectroscopy (LIBS) Combined with Temporal Plasma Analysis of C2 Molecular Emission for Carbon Analysis in Coal

A benchtop Laser-Induced Breakdown Spectroscopy (LIBS) is demonstrated to determine the elemental carbon content present in raw coal used for combustion in power plants. The spectral intensities of molecular CN and C2 emission are measured together with the atomic carbon (C) and other inorganic elements (Si, Fe, Mg, Al, Ca, Na, and K) in the LIBS spectrum of coal. The emission persistence time of C2 molecule emission is measured from the coal plasma generated by a nanosecond laser ablation with a wavelength of 266 nm in the Ar atmosphere. The emission persistence time of molecular C2 emission along with the spectral intensities of major ash elements (Fe, Si, Al, and Ca) and carbon emissions (atomic C, molecular CN, and C2) shows a better relationship with the carbon wt% of different coal samples. The calibration model to measure elemental carbon (wt%) is developed by combining the spectral characteristics (Spectral intensity) and the temporal characteristics (Emission persistence time of C2 molecule emission). The temporal characteristic studies combined with the spectroscopic data in the PLSR (Partial Least Square Regression) model has resulted in an improvement in the root mean square error of validation (RMSEV), and the relative standard deviation (RSD) is reduced from 10.86% to 4.12% and from 11.32% to 6.04%, respectively.

Спектроскопия барьерного разряда низкого давления. Послесвечение с ионами Ne-=SUP=-2+-=/SUP=-, Ne-=SUP=-+-=/SUP=- и Ne-=SUP=-2+-=/SUP=-

We present the results of modeling the radiation of a decaying plasma, formed by the processes of electron-ion recombination with the participation of three neon ions: the molecular ion Ne2+ and atomic ions Ne+ and Ne2+. Such a combination of ions, simultaneously participating in the formation of the plasma spectrum, was first discovered in the afterglow of a pulsed barrier discharge of a cylindrical geometry at neon pressures less than 1 Torr and an electron density[e] ≤ 4 × 1010 cm-3. The main attention is paid to the comparative analysis of the mechanisms of impact-radiation recombination of Ne+ and Ne2+ ions based on the numerical solution of the system of differential equations for the densities of ions and long-lived excited atoms in the afterglow, taking into account the main elementary processes in decaying plasma with pulsed "heating" of electrons. The regularities of electron temperature relaxation from discharge values of several electron volts to 300 K in the late afterglow are considered in particular details. Comparison of the model solutions with the spectral intensities measured by the multichannel photon counting method shows that, given their good agreement in the case of singly charged ions, an adequate description of the evolution of ionic lines requires expanding the available information on the recombination of Ne2+ ions.

Chemometrics-Enabled Raman Spectrometric Qualitative Determination and Assessment of Biochemical Alterations during Early Prostate Cancer Proliferation in Model Tissue

The use of Raman spectroscopy combined with multivariate chemometrics for disease diagnosis has attracted great attention from researchers in recent years. This is because it is a noninvasive and nondestructive detection approach with enhanced sensitivity. However, a major challenge when analyzing spectra from biological samples has been the detection of subtle biochemical alterations buried in background and fluorescence noise. This work reports a qualitative chemometrics-assisted investigation of subtle biochemical alterations associated with prostate malignancy in model biological tissue (metastatic androgen insensitive (PC3) and immortalized normal (PNT1a) prostate cell lines). Raman spectra were acquired from PC3 and PNT1a cells at various stages of growth, and their biochemical alterations were determined from difference spectra between the two cell lines (for prominent alterations) and principal component analysis (PCA) (for subtle alterations). The Raman difference spectra were computed by subtracting the normalized mean spectral intensities of PNT1a cells from the normalized mean spectral intensities of PC3 cells. These difference spectra revealed prominent biochemical alterations associated with the malignant PC3 cells at 566 ± 0.70 cm−1, 630 cm−1, 1370 ± 0.86 cm−1, and 1618 ± 1.73 cm−1 bands. The band intensity ratios at 566 ± 0.70 cm−1 and 630 cm−1 suggested that prostate malignancy can be associated with an increase in relative amounts of nucleic acids and lipids, respectively, whereas those at 1370 ± 0.86 cm−1 and 1618 ± 1.73 cm−1 suggested that prostate malignancy can be associated with a decrease in relative amounts of saccharides and tryptophan, respectively. In the analysis using PCA, intermediate-order and high-order principal components (PCs) were used to extract the subtle biochemical fingerprints associated with the cell lines. This revealed subtle biochemical differences at 1076 cm−1, (1232, 1234 cm−1), (1276, 1278 cm−1), (1330, 1333 cm−1), (1434, 1442 cm−1), and (1471, 1479 cm−1). The band intensity ratios at 1076 cm−1 and 1232 cm−1 suggested that prostate malignancy can be associated with an increase in subtle amounts of nucleic acids and amide III components, respectively. The method reported here has demonstrated that subtle biochemical alterations can be extracted from Raman spectra of normal and malignant cell lines. The identified subtle bands could play an important role in quantitative monitoring of early biomarker alterations associated with prostate cancer proliferation.

Changes in molecular structure of protein and carbohydrate in soybean products with different processing methods and their effects on nutrient degradation characteristics of the products

In this study, four types of soybean products with different processing methods (soybean meal 1 and soybean meal 2, extruded soybean meal, fermented soybean meal and extruded soybean) were used to examine the effect of fermentation and extrusion on molecular structures of protein and carbohydrate. Extrusion and fermentation significantly decreased (P < 0.05) the values of related protein spectral intensities (height and area of amide and secondary structure) and the biggest reduction was found in extruded soybean compared to soybean meal 1 and soybean meal 2. Compared with extruded soybean meal, the area ratio of amide I to amide II and the height ratio of α-helix to β sheet in extruded soybean were significantly reduced (P < 0.05), and there was no difference in these spectral values between extruded and fermented soybean. Extrusion and fermentation significantly decreased (P < 0.05) the values of carbohydrate spectral intensities, including structural carbohydrate (STCHO) and cellulosic compounds (CELC) and total carbohydrate (CHO), compared to soybean meal 2. The ratio of α-helix to β-sheet was positively related to the DM of soybean degradability in the rumen (P < 0.05, r = 0.590), so was A-CELC to A-STCHO (P < 0.05, r = 0.747). A positive relationship was found between CP degradability in the rumen and the area ratios of amide I and amide II, CELC to CHO, and STCHO to CHO. Spectral intensity of CHO area was negatively associated with neutral detergent fibre (NDF) and acid detergent fibre (ADF) degradability in the rumen. The study indicated that extrusion and fermentation could alter the molecular structure of protein and carbohydrate and the degradation characteristics of soybean products in the rumen.

FULL-SCALE STUDIES OF FLUORESCENCE FEATURES OF SURFACTANT FILMS ON EUTROPHIC WATER WAVY SURFACE

The current paper presents the results of a full-scale study of the fluorescence features of pollutants ( diesel fuel, Urals oil), as well as an alcoholic solution of oleic acid, which imitates oil by visco-elastic characteristics. The experiment was carried out on the Gorky Reservoir under conditions of wind waves. Local areas of artificial slicks were created for multiple intersections at different angles to the wind and waves with continuous lidar sounding of the upper water layer. Signals were recorded by the ninth generation UVL-9 laser ultraviolet fluorescent lidar developed by the Institute of Oceanology of the Russian Academy of Sciences. At signal analysis, spectral intensities and fluorescence features of the studied substances are determined, possibilities of slick detecting as well as film thickness estimation are made, examples of surfactant film detecting behind the oleic slick where visual observation of pollutants does not give a result are showed. The obtained results are useful for the further development of fluorescent methods and devices for operational remote sensing of surface pollution on wave water surface.

Spurious Absorption Frequency Appearance Due to Frequency Conversion Processes in Pulsed THz TDS Problems

The appearance of the spurious absorption frequencies caused by the frequency conversion process at the broadband THz pulse propagation in a medium is theoretically and experimentally discussed. The spurious absorption frequencies appear due to both the frequency doubling and generation of waves with sum or difference frequency. Such generation might occur because of the nonlinear response of a medium or its non-instantaneous response. This phenomenon is confirmed by the results of a few physical experiments provided with the THz CW signals and broadband THz pulses that are transmitted through the ordinary or dangerous substances. A high correlation between the time-dependent spectral intensities for the basic frequency and generated frequencies is demonstrated while using the computer simulation results. This feature of the frequency conversion might be used for the detection and identification of a substance.

Spectroscopic Examination, DFT Calculations, Electronic and Optical Properties of α-santonin

In this work, the FTIR and FT-Raman spectra of α-santonin have been recorded and vibrational analysis of the compound were carried out from the observed data. The molecular parameters such as bond length, bond angle, frequencies and spectral intensities were estimated with the help of density functional theory (DFT/B3LYP) method. The calculated and experimental spectra were compared and the stability of the molecule has been studied by the natural bond orbital analysis. The electronic properties like HOMO-LUMO have also been analyzed. The nonlinear optical properties of the molecule have been analyzed by calculating the first hyperpolarisability and comparing with urea.