Surface‐enhanced Raman spectroscopy (SERS) is a unique technique to study submembrane hemoglobin (Hbsm) in erythrocytes. We report the detailed design of SERS experiments on living erythrocytes to estimate dependence of the enhancemen t factor for main Raman bands of Hbsm on silver nanoparticle (AgNP) properties. We demonstrate that the enhancement factor for ν 4/A1g, ν 10/B1g and A2g Raman bands of Hbsm varies from 105 to 107 under proposed experimental conditions with 473 nm laser excitation. For the first time we show that the enhancement of Raman scattering increases with the increase in the relative amount of small NPs in colloids, with the decrease in AgNP size and with plasmon resonance shift to the shorter wavelength region. Obtained results can be explained by the ability of smaller AgNPs to get deeper into nano‐invaginations of the plasma membrane than larger AgNPs. This shortens the distance between small AgNPs and Hbsm and, consequently, leads to the higher enhancement of Raman scattering of Hbsm. The enhancement of higher wavenumber bands ν 10/B1g and A2g is more sensitive to AgNPs’ size and the relative amount of small AgNPs than the enhancement of the lower wavenumber band ν 4/A1g. This can be used for AgNP‐controlled enhancement of the desired Raman bands and should be taken into account in biomedical SERS experiments. Copyright © 2013 John Wiley & Sons, Ltd.
Nowadays, there is an interest in biomedical and nanobiotechnological studies, such as studies on carotenoids as antioxidants and studies on molecular markers for cardiovascular, endocrine, and oncological diseases. Moreover, interest in industrial production of microalgal biomass for biofuels and bioproducts has stimulated studies on microalgal physiology and mechanisms of synthesis and accumulation of valuable biomolecules in algal cells. Biomolecules such as neutral lipids and carotenoids are being actively explored by the biotechnology community. Raman spectroscopy (RS) has become an important tool for researchers to understand biological processes at the cellular level in medicine and biotechnology. This review provides a brief analysis of existing studies on the application of RS for investigation of biological, medical, analytical, photosynthetic, and algal research, particularly to understand how the technique can be used for lipids, carotenoids, and cellular research. First, the review article shows the main applications of the modified Raman spectroscopy in medicine and biotechnology. Research works in the field of medicine and biotechnology are analysed in terms of showing the common connections of some studies as caretenoids and lipids. Second, this article summarises some of the recent advances in Raman microspectroscopy applications in areas related to microalgal detection. Strategies based on Raman spectroscopy provide potential for biochemical-composition analysis and imaging of living microalgal cells, in situ and in vivo. Finally, current approaches used in the papers presented show the advantages, perspectives, and other essential specifics of the method applied to plants and other species/objects.
The neuroprotective effect of the natural antioxidant taxifolin (TAX) is well known for ischemic pathologies. However, the limitations of taxifolin application are described—poor solubility, low ability to penetrate the blood–brain barrier, and side effects from high doses for stroke therapy. We proposed the problem of targeted delivery of taxifolin and achievement effective concentrations could be solved by developing a nanocomplex of selenium nanoparticles (SeNPs) with taxifolin (Se–TAX). In this study, we developed a selenium–taxifolin nanocomplex based on selenium nanoparticles with a 100 nm size. It was shown that TAX, SeNPs, and Se–TAX were all able to suppress the production of ROS in neurons and astrocytes under exposure to exogenous H2O2 and ischemia-like conditions. However, the Se–TAX nanocomplex appeared to be the most effective, displaying a lower working concentration range and negligible pro-oxidant effect compared with pure SeNPs. The mechanism of Se–TAX beneficial effects involved the activation of some antioxidant enzymes and the suppression of ROS-generating systems during OGD/reoxygenation, while TAX and “naked” SeNPs were less effective in regulating the cellular redox status. Naked SeNPs inhibited a global increase in Ca2+ ions in cytosol, but not OGD-induced hyperexcitation of the neuroglial network, while Se–TAX suppressed both [Ca2+]i rise and hyperexcitation. The effect of TAX at similar doses appeared exclusively in inhibiting OGD-induced hyperexcitation. Analysis of necrosis and apoptosis after OGD/reoxygenation revealed the highest efficiency of the Se–TAX nanocomplex as well. Se–TAX suppressed the expression of proinflammatory and proapoptotic proteins with simultaneous activation of protective genes. We conclude that the Se–TAX nanocomplex combines the antioxidative features taxifolin and the antiapoptotic effect of nanoselenium, involving the regulation of Ca2+ dynamics.
Despite the fact that sorafenib is recommended for the treatment of oncological diseases of the liver, kidneys, and thyroid gland, and recently it has been used for combination therapy of brain cancer of various genesis, there are still significant problems for its widespread and effective use. Among these problems, the presence of the blood–brain barrier of the brain and the need to use high doses of sorafenib, the existence of mechanisms for the redistribution of sorafenib and its release in the brain tissue, as well as the high resistance of gliomas and glioblastomas to therapy should be considered the main ones. Therefore, there is a need to create new methods for delivering sorafenib to brain tumors, enhancing the therapeutic potential of sorafenib and reducing the cytotoxic effects of active compounds on the healthy environment of tumors, and ideally, increasing the survival of healthy cells during therapy. Using vitality tests, fluorescence microscopy, and molecular biology methods, we showed that the selenium-sorafenib (SeSo) nanocomplex, at relatively low concentrations, is able to bypass the mechanisms of glioblastoma cell chemoresistance and to induce apoptosis through Ca2+-dependent induction of endoplasmic reticulum stress, changes in the expression of selenoproteins and selenium-containing proteins, as well as key kinases-regulators of oncogenicity and cell death. Selenium nanoparticles (SeNPs) also have a high anticancer efficacy in glioblastomas, but are less selective, since SeSo in cortical astrocytes causes a more pronounced activation of the cytoprotective pathways.
Neonicotinoid insecticides are used against the wide range of pests to protect plants. The influence of neonicotinoids on target and non-target insects is well understood. Hence, there are controversial opinions about the effect of neonicotinoids on the plants. We investigated pigments and photosynthetic primary reactions in two maize genotypes (the inbred line zppl 225 and hybrid zp 341) under thiamethoxam (TMX) treatment by root irrigation. It was found that the effect of TMX depended on pesticide application techniques and selection of maize genotype. TMX was added to the soil by root irrigation on the 4th and 8th days after planting, and photosynthetic characteristics monitored on the 10th and 12th days after planting. The primary photochemical reactions in PSII (Fv/Fm) of both maize genotypes were not affected under two variants of TMX treatment during all growing period. The hybrid zp341 was shown to be more susceptible to both TMX treatments, demonstrating a decrease in photosynthetic characteristics (JIP-test parameters) as well as changes in the content of pigments and in the conformation of the carotenoid molecule. Our findings suggest that the combination of fluorescence method and Raman spectroscopy is a perspective tool for monitoring plant state under pesticide application.
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