Detection of carbon nanotubes in bovine raw milk through Fourier transform Raman spectroscopy.

The potential use of carbon-based methodologies for drug delivery and reproductive biology in cows raises concerns about residues in milk and food safety. This study aimed to assess the potential of Fourier transform Raman spectroscopy and discriminant analysis using partial least squares (PLS-DA) to detect functionalized multiwalled carbon nanotubes (MWCNT) in bovine raw milk. Oxidized MWCNT were diluted in milk at different concentrations from 25.00 to 0.01 µg/mL. Raman spectroscopy measurements and PLS-DA were performed to identify low concentrations of MWCNT in milk samples. The PLS-DA model was characterized by the analysis of the variable importance in projection (VIP) scores. All the training samples were correctly classified by the model, resulting in no false-positive or false-negative classifications. For test samples, only one false-negative result was observed, for 0.01 µg/mL MWCNT dilution. The association between Raman spectroscopy and PLS-DA was able to identify MWCNT diluted in milk samples up to 0.1 µg/mL. The PLS-DA model was built and validated using a set of test samples and spectrally interpreted based on the highest VIP scores. This allowed the identification of the vibrational modes associated with the D and G bands of MWCNT, as well as the milk bands, which were the most important variables in this analysis.

Unraveling surface-enhanced Raman spectroscopy results through chemometrics and machine learning: principles, progress, and trends.

Surface-enhanced Raman spectroscopy (SERS) has gained increasing attention because it provides rich chemical information and high sensitivity, being applicable in many scientific fields including medical diagnosis, forensic analysis, food control, and microbiology. Although SERS is often limited by the lack of selectivity in the analysis of samples with complex matrices, the use of multivariate statistics and mathematical tools has been demonstrated to be an efficient strategy to circumvent this issue. Importantly, since the rapid development of artificial intelligence has been promoting the implementation of a wide variety of advanced multivariate methods in SERS, a discussion about the extent of their synergy and possible standardization becomes necessary. This critical review comprises the principles, advantages, and limitations of coupling SERS with chemometrics and machine learning for both qualitative and quantitative analytical applications. Recent advances and trends in combining SERS with uncommonly used but powerful data analysis tools are also discussed. Finally, a section on benchmarking and tips for selecting the suitable chemometric/machine learning method is included. We believe this will help to move SERS from an alternative detection strategy to a general analytical technique for real-life applications.

Determination of amylose content in starch using Raman spectroscopy and multivariate calibration analysis.

Fourier transform Raman spectroscopy and chemometric tools have been used for exploratory analysis of pure corn and cassava starch samples and mixtures of both starches, as well as for the quantification of amylose content in corn and cassava starch samples. The exploratory analysis using principal component analysis shows that two natural groups of similar samples can be obtained, according to the amylose content, and consequently the botanical origins. The Raman band at 480 cm(-1), assigned to the ring vibration of starches, has the major contribution to the separation of the corn and cassava starch samples. This region was used as a marker to identify the presence of starch in different samples, as well as to characterize amylose and amylopectin. Two calibration models were developed based on partial least squares regression involving pure corn and cassava, and a third model with both starch samples was also built; the results were compared with the results of the standard colorimetric method. The samples were separated into two groups of calibration and validation by employing the Kennard-Stone algorithm and the optimum number of latent variables was chosen by the root mean square error of cross-validation obtained from the calibration set by internal validation (leave one out). The performance of each model was evaluated by the root mean square errors of calibration and prediction, and the results obtained indicate that Fourier transform Raman spectroscopy can be used for rapid determination of apparent amylose in starch samples with prediction errors similar to those of the standard method.

Spectroscopic and theoretical study of the "azo"-dye E124 in condensate phase: evidence of a dominant hydrazo form.

Spectroscopic techniques, including Raman, IR, UV/vis, and NMR were used to characterize the samples of the azo dye Ponceau 4R (also known as E124, New Coccine; Cochineal Red; C.I. no. 16255; Food Red No. 102), which is 1,3-naphthalenedisulfonic acid, 7-hydroxy-8-[(4-sulfo-1-naphthalenyl) azo] trisodium salt in aqueous solution and solid state. In addition, first principle calculations were carried out for the azo (OH) and hydrazo (NH) tautomers in order to assist in the assignment of the experimental data. The two intense bands observed in the UV/vis spectrum, centered at 332 and 507 nm, can be compared to the calculated values at 296 and 474 nm for azo and 315 and 500 nm for hydrazo isomer, with the latter in closer agreement to the experiment. The Raman spectrum is quite sensitive to tautomeric equilibrium; in solid state and aqueous solution, three bands were observed around 1574, 1515, and 1364 cm(-1), assigned to mixed modes including deltaNH + betaCH + nuCC, deltaNH + nuC horizontal lineO + nuC horizontal lineN + betaCH and nuCC vibrations, respectively. These assignments are predicted only for the NH species centered at 1606, 1554, and 1375 cm(-1). The calculated Raman spectrum for the azo (OH) tautomer showed two strong bands at 1468 (nuN = N + deltaOH) and 1324 cm(-1) (nuCC + nuC-N), which were not obtained experimentally. The (13)C NMR spectrum showed a very characteristic peak at 192 ppm assigned to the carbon bound to oxygen in the naphthol ring; the predicted values were 165 ppm for OH and 187 for NH isomer, supporting once again the predominance of NH species in solution. Therefore, all of the experimental and theoretical results strongly suggest the food dye Ponceau 4R or E124 has a major contribution of the hydrazo structure instead of the azo form as the most abundant in condensate phase.

Physics, chemistry, and Hirshfeld surface analyses of gamma-irradiated thalidomide to evaluate behavior under sterilization doses.

Thalidomide was indicated as a sedative and antiemetic and prescribed for pregnant women. Its tragic teratogenic effects culminated in withdrawal from the market. Since the discovery of its anti-angiogenic and anti-inflammatory actions, thalidomide has been used in the treatment of leprosy and multiple myeloma, which justify studies of its stability. We investigated the effects of irradiation of thalidomide up to 100 kGy (fourfold the usual sterilizing dose for pharmaceutics). The ß polymorph of thalidomide was obtained in an isothermal experiment at 270 °C. All samples underwent gamma irradiation for specific times. At different doses, decomposition of the pharmaceutical was not observed up to 100 kGy. The observed effect was angle turning between the phthalimide and glutarimide rings modulated by repulsion towards the carbonyl group, leading to a stable energetic configuration, as measured by the equilibrium in the torsion angle after irradiation. The thalidomide molecule has a center of symmetry, so a full turn starting from 57.3° will lead to an identical molecule. Further irradiation will start the process again. Samples irradiated at 30 and 100 kGy have more compact unit cells and a lower volume, which leads to an increase in the intermolecular hydrogen interaction within the unit cell, resulting in higher thermal stability for polymorph α.

Natural Schistosoma mansoni Infection in the Wild Reservoir Nectomys squamipes Leads to Excessive Lipid Droplet Accumulation in Hepatocytes in the Absence of Liver Functional Impairment.

Schistosomiasis is a neglected tropical disease of a significant public health impact. The water rat Nectomys squamipes is one of the most important non-human hosts in the schistosomiasis mansoni transmission in Brazil, being considered a wild reservoir. Cellular mechanisms that contribute to the physiological adaptation of this rodent to the Schistosoma mansoni parasite are poorly understood. Here we identified, for the first time, that a hepatic steatosis, a condition characterized by excessive lipid accumulation with formation of lipid droplets (LDs) within hepatocytes, occurs in response to the natural S. mansoni infection of N. squamipes, captured in an endemic region. Significant increases of LD area in the hepatic tissue and LD numbers/hepatocyte, detected by quantitative histopathological and ultrastructural analyses, were paralleled by increased serum profile (total cholesterol and triglycerides) in infected compared to uninfected animals. Raman spectroscopy showed high content of polyunsaturated fatty acids (PUFAs) in the liver of both groups. MALDI-TOFF mass spectroscopy revealed an amplified pool of omega-6 PUFA arachidonic acid in the liver of infected animals. Assessment of liver functional activity by the levels of hepatic transaminases (ALT and AST) did not detect any alteration during the natural infection. In summary, this work demonstrates that the natural infection of the wild reservoir N. squamipes with S. mansoni elicits hepatic steatosis in the absence of liver functional harm and that accumulation of lipids, markedly PUFAs, coexists with low occurrence of inflammatory granulomatous processes, suggesting that lipid stores may be acting as a protective mechanism for dealing with the infection.

Lipid Body Organelles within the Parasite Trypanosoma cruzi: A Role for Intracellular Arachidonic Acid Metabolism.

Most eukaryotic cells contain varying amounts of cytosolic lipidic inclusions termed lipid bodies (LBs) or lipid droplets (LDs). In mammalian cells, such as macrophages, these lipid-rich organelles are formed in response to host-pathogen interaction during infectious diseases and are sites for biosynthesis of arachidonic acid (AA)-derived inflammatory mediators (eicosanoids). Less clear are the functions of LBs in pathogenic lower eukaryotes. In this study, we demonstrated that LBs, visualized by light microscopy with different probes and transmission electron microscopy (TEM), are produced in trypomastigote forms of the parasite Trypanosoma cruzi, the causal agent of Chagas' disease, after both host interaction and exogenous AA stimulation. Quantitative TEM revealed that LBs from amastigotes, the intracellular forms of the parasite, growing in vivo have increased size and electron-density compared to LBs from amastigotes living in vitro. AA-stimulated trypomastigotes released high amounts of prostaglandin E2 (PGE2) and showed PGE2 synthase expression. Raman spectroscopy demonstrated increased unsaturated lipid content and AA incorporation in stimulated parasites. Moreover, both Raman and MALDI mass spectroscopy revealed increased AA content in LBs purified from AA-stimulated parasites compared to LBs from unstimulated group. By using a specific technique for eicosanoid detection, we immunolocalized PGE2 within LBs from AA-stimulated trypomastigotes. Altogether, our findings demonstrate that LBs from the parasite Trypanosoma cruzi are not just lipid storage inclusions but dynamic organelles, able to respond to host interaction and inflammatory events and involved in the AA metabolism. Acting as sources of PGE2, a potent immunomodulatory lipid mediator that inhibits many aspects of innate and adaptive immunity, newly-formed parasite LBs may be implicated with the pathogen survival in its host.

Detection of explosives on the surface of banknotes by Raman hyperspectral imaging and independent component analysis.

The aim of this study was to develop a methodology using Raman hyperspectral imaging and chemometric methods for identification of pre- and post-blast explosive residues on banknote surfaces. The explosives studied were of military, commercial and propellant uses. After the acquisition of the hyperspectral imaging, independent component analysis (ICA) was applied to extract the pure spectra and the distribution of the corresponding image constituents. The performance of the methodology was evaluated by the explained variance and the lack of fit of the models, by comparing the ICA recovered spectra with the reference spectra using correlation coefficients and by the presence of rotational ambiguity in the ICA solutions. The methodology was applied to forensic samples to solve an automated teller machine explosion case. Independent component analysis proved to be a suitable method of resolving curves, achieving equivalent performance with the multivariate curve resolution with alternating least squares (MCR-ALS) method. At low concentrations, MCR-ALS presents some limitations, as it did not provide the correct solution. The detection limit of the methodology presented in this study was 50 µg cm(-2).

Increasing teachers' knowledge about ADHD and learning disorders: an investigation on the role of a psychoeducational intervention.

OBJECTIVE: To investigate elementary school teachers' baseline knowledge about ADHD and learning disorders (LD) and the impact of a strategy to increase awareness of these disorders. METHOD: A total of 37 teachers were selected from four elementary schools in the catchment area of the University Hospital, in Porto Alegre, Brazil. To evaluate teachers' knowledge, two self-report questionnaires about ADHD and LD were applied before and after an awareness program on these disorders. RESULTS: The intervention significantly increased teachers' knowledge of both disorders, even after adjustment for confounding factors (p < .001). In the repeated measures ANCOVA, only teachers' previous knowledge of ADHD/LD (p < .001) was significant in predicting score change in knowledge before and after the intervention. CONCLUSION: Results suggest the efficacy of a brief psychoeducational intervention program for increasing teacher awareness and knowledge about ADHD and LD. Future studies are warranted to confirm the efficacy and evaluate the long-term impact of this intervention.

Classification of Amazonian rosewood essential oil by Raman spectroscopy and PLS-DA with reliability estimation.

The Amazon tree Aniba rosaeodora Ducke (rosewood) provides an essential oil valuable for the perfume industry, but after decades of predatory extraction it is at risk of extinction. The extraction of the essential oil from wood implies the cutting of the tree, and then the study of oil extracted from the leaves is important as a sustainable alternative. The goal of this study was to test the applicability of Raman spectroscopy and Partial Least Square Discriminant Analysis (PLS-DA) as means to classify the essential oil extracted from different parties (wood, leaves and branches) of the Brazilian tree A. rosaeodora. For the development of classification models, the Raman spectra were split into two sets: training and test. The value of the limit that separates the classes was calculated based on the distribution of samples of training. This value was calculated in a manner that the classes are divided with a lower probability of incorrect classification for future estimates. The best model presented sensitivity and specificity of 100%, predictive accuracy and efficiency of 100%. These results give an overall vision of the behavior of the model, but do not give information about individual samples; in this case, the confidence interval for each sample of classification was also calculated using the resampling bootstrap technique. The methodology developed have the potential to be an alternative for standard procedures used for oil analysis and it can be employed as screening method, since it is fast, non-destructive and robust.

Rapid assessment of chemical compounds from Phyllogorgia dilatata using Raman spectroscopy

Abstract The gorgonian Phyllogorgia dilatata is endemic to the Brazilian coast which is listed as threatened with extinction. This species is known to produce sterols, mono- to tetra-terpenes, conjugated polyenals and peptides. The main objective of this study is to present an alternative method for identification of different classes of compounds based upon a Raman mapping technique using FT-Raman spectroscopy. The Raman analysis performed directly on the tissues (in situ) revealed the occurrence of peridinin, diadinoxanthin, conjugated polyenal and linoleic acid, that were also confirmed by Raman analysis of partitioned crude extracts. We have demonstrated that the technique has potential for use in guiding chromatographic separations and in providing information with respect to the early stages of a tissue necrosis through “purpling”. It may become a valuable non-destructive technique for monitoring the accumulation or production of metabolites during a biological interaction.