In vivo study of dermal collagen of striae distensae by confocal Raman spectroscopy.

This research work mainly deals with studying qualitatively the changes in the dermal collagen of two forms of striae distensae (SD) namely striae rubrae (SR) and striae albae (SA) when compared to normal skin (NS) using confocal Raman spectroscopy. The methodology includes an in vivo human skin study for the comparison of confocal Raman spectra of dermis region of SR, SA, and NS by supervised multivariate analysis using partial least squares discriminant analysis (PLS-DA) to determine qualitatively the changes in dermal collagen. These groups are further analyzed for the extent of hydration of dermal collagen by studying the changes in the water content bound to it. PLS-DA score plot showed good separation of the confocal Raman spectra of dermis region into SR, SA, and NS data groups. Further analysis using loading plot and S-plot indicated the participation of various components of dermal collagen in the separation of these groups. Bound water content analysis showed that the extent of hydration of collagen is more in SD when compared to NS. Based on the results obtained, this study confirms the active involvement of dermal collagen in the formation of SD. It also emphasizes the need to study quantitatively the role of these various biochemical changes in the dermal collagen responsible for the variance between SR, SA, and NS.

In Vivo Human Skin Penetration Study of Sunscreens by Confocal Raman Spectroscopy.

This research work mainly deals with the application of confocal Raman spectroscopic technique to study in vivo human skin penetration of sunscreen products, as there are a lot of controversies associated with their skin penetration. Healthy human volunteers were tested for penetration of two commercial sunscreen products into their volar forearm skin for a period of 2 h. Measurements were taken before and after application of these sunscreen products. All the confocal Raman spectra were pre-processed and then subjected to multivariate two-dimensional principal component analysis and classical least squares analysis to determine the skin penetration of these sunscreens in comparison to the "sunscreen product spectrum" which was considered as the control. Score plots of principal component analysis of confocal Raman spectra indicated clear separation between the spectra before and after application of sunscreen products. Loading plots showed the maximum differences in the spectral region from 1590 to 1626 cm-1 where the characteristic peak of the pure sunscreen products was observed. Classical least squares analysis has shown a significant penetration to a depth of 10 µm in the volar forearm skin of healthy human volunteers for both these sunscreen products. The results confirm that the penetration of these tested sunscreen products was restricted to stratum corneum and also prove that confocal Raman spectroscopy is a simple, fast, nondestructive, and noninvasive semi-quantitative analytical technique for these studies.

Ketamine can be produced by Pochonia chlamydosporia: an old molecule and a new anthelmintic?

BACKGROUND: Infection by nematodes is a problem for human health, livestock, and agriculture, as it causes deficits in host health, increases production costs, and incurs a reduced food supply. The control of these parasites is usually done using anthelmintics, which, in most cases, have not been fully effective. Therefore, the search for new molecules with anthelmintic potential is necessary. METHODS: In the present study, we isolated and characterized molecules from the nematophagous fungus Pochonia chlamydosporia and tested these compounds on three nematodes: Caenorhabditis elegans; Ancylostoma ceylanicum; and Ascaris suum. RESULTS: The ethyl acetate extract showed nematicidal activity on the nematode model C. elegans. We identified the major substance present in two sub-fractions of this extract as ketamine. Then, we tested this compound on C. elegans and the parasites A. ceylanicum and A. suum using hamsters and mice as hosts, respectively. We did not find a difference between the animal groups when considering the number of worms recovered from the intestines of animals treated with ketamine (6 mg) and albendazole (P > 0.05). The parasite burden of larvae recovered from the lungs of mice treated with ketamine was similar to those treated with ivermectin. CONCLUSIONS: The results presented here demonstrate the nematicidal activity of ketamine in vitro and in vivo, thus confirming the nematicidal potential of the molecule present in the fungus P. chlamydosporia may consist of a new method of controlling parasites.

Discrimination between conventional and omega-3 fatty acids enriched eggs by FT-Raman spectroscopy and chemometric tools.

This work developed an analytical method to differentiate conventional and omega-3 fat acids enriched eggs by Raman spectroscopy and multivariate supervised classification with Partial Least Squares Discriminant Analysis (PLS-DA). Forty samples of enriched eggs and forty samples of different types of common eggs from different batches were used to build the model. Firstly, gas chromatography was employed to analyze fatty acid profiles in egg samples. Raman spectra of the yolk extracts were recorded in the range from 3100 to 990 cm-1. PLS-DA model was able to correctly classify samples with nearly 100% success rate. This model was validated estimating appropriate figures of merit. Predictions uncertainties were also estimated by bootstrap resampling. The most discriminant Raman modes were identified based on VIP (variables importance in projection) scores. This method has potential to assist food industries and regulatory agencies for food quality control, allowing detecting frauds and enabling faster and reliable analyzes.

In vivo confocal Raman spectroscopy for intrinsic aging and photoaging assessment.

BACKGROUND: In vivo confocal Raman spectroscopy is a non-invasive method to assess either the epidermis or the dermis composition. Few studies have focused on dermis collagen alterations through intrinsic aging and photoaging. OBJECTIVE: This study evaluated the in vivo Raman spectra from the dermis of a photoexposed site versus a non-photoexposed region in different age groups, and evaluated the correlation between peak intensities and age, photoaging score and the amount of collagen assessed with histology and high frequency ultrasound (HFUS). METHODS: Fifteen volunteers aged 28-82 years were divided into three groups according to forearm photoaging degree. In vivo Raman spectra from the dermis were collected on the dorsal forearm (chronically photoexposed skin) and on the proximal medial arm (non-photoexposed skin). Cross-sectional images of the skin were obtained using a 20MHz ultrasound unit exactly on the same sites, which were further submitted to punch biopsies for histologic study (collagen I immunohistochemistry, picrosirius red staining and Verhoeff). Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) were taken in the spectral region of 796cm-1-996cm-1 to determine its potential to discriminate between different groups. The Spearman rank correlation coefficient of individual peak intensities and ratios with age, clinical score and the amount of collagen assessed by ultrasound and histology were calculated. RESULTS: PCA of pairs of groups and OPLS-DA could discriminate the intrinsically from the photoaged skin and the young group from the elderly one, with important contribution of the 938cm-1 and 855cm-1 peaks intensities. The intensity of the peaks in 855cm-1 and/or 938cm-1 presented moderate correlation with age (rho=0.579, p=0.049) and moderate to high inverse correlation with HFUS echogenicity (rho=-0.710, p=0.010) and collagen I immunohistochemistry (rho=-0.833, p=0.005) in the non-photoexposed region. The I1275/I1450 intensities ratio presented moderate to high correlation coefficients with age (rho=-0.730, p=0.007), photoaging score (rho=-0.594, p=0.042), HFUS echogenicity (rho=0.760, p<0.001) and histology (collagen I immunohistochemistry (rho=0.643, p=0.024), picrosirius (rho=0.773, p=0.005) and Verhoeff (rho=-0.727, p=0.011)) in the photoexposed site. CONCLUSION: The wavenumber region between 798 and 994cm-1 is useful for the analysis of dermal collagen alterations through the intrinsic aging process, while photoaging is better assessed by the I1275/I1450 intensities ratio. This is the first skin aging study to show a correlation between Raman peaks and the amount of collagen assessed by HFUS and histology.

Capillary zone electrophoresis for fatty acids with chemometrics for the determination of milk adulteration by whey addition.

Adulteration of milk with whey is difficult to detect because these two have similar physical and chemical characteristics. The traditional methodologies to monitor this fraud are based on the analysis of caseinomacropeptide. The present study proposes a new approach to detect and quantify this fraud using the fatty acid profiles of milk and whey. Fatty acids C14:0, C16:0, C18:0, C18:1, C18:2 and C18:3 were selected by gas chromatography associated with discriminant analysis to differentiate milk and whey, as they are present in quite different amounts. These six fatty acids were quantified within a short time by capillary zone electrophoresis in a set of adulterated milk samples. The correlation coefficient between the true values of whey addition and the experimental values obtained by this technique was 0.973. The technique is thus useful for the evaluation of milk adulteration with whey, contributing to the quality control of milk in the dairy industry.