Raman Scattering:From Structural Biology to Medical Applications

This is a review of relevant Raman spectroscopy (RS) techniques and their use in structural biology, biophysics, cells, and tissues imaging towards development of various medical diagnostic tools, drug design, and other medical applications. Classical and contemporary structural studies of different water-soluble and membrane proteins, DNA, RNA, and their interactions and behavior in different systems were analyzed in terms of applicability of RS techniques and their complementarity to other corresponding methods. We show that RS is a powerful method that links the fundamental structural biology and its medical applications in cancer, cardiovascular, neurodegenerative, atherosclerotic, and other diseases. In particular, the key roles of RS in modern technologies of structure-based drug design are the detection and imaging of membrane protein microcrystals with the help of coherent anti-Stokes Raman scattering (CARS), which would help to further the development of protein structural crystallography and would result in a number of novel high-resolution structures of membrane proteins—drug targets; and, structural studies of photoactive membrane proteins (rhodopsins, photoreceptors, etc.) for the development of new optogenetic tools. Physical background and biomedical applications of spontaneous, stimulated, resonant, and surface- and tip-enhanced RS are also discussed. All of these techniques have been extensively developed during recent several decades. A number of interesting applications of CARS, resonant, and surface-enhanced Raman spectroscopy methods are also discussed

Gentamicin sulphate permeation through porcine intestinal epithelial cell monolayer

Gentamicin is an aminoglycoside antibiotic widely used in combination with dimethyl sulphoxide (DMSO) in topical drug formulations. It is not known, however, whether DMSO can enhance the permeation of gentamicin through biological membranes, leading to oto- and nephrotoxic side effects. A simple and reliable high-performance liquid chromatographic (HPLC) method was applied for the quantitative determination of gentamicin collected from the apical and basolateral compartments of the porcine intestinal epithelial cell line IPEC-J2 cell monolayer using fluorometric derivatisation of the analyte with fluorenylmethyloxycarbonyl chloride (FMOC) prior to chromatographic run in the presence and absence of 1% DMSO. The lack of change in transepithelial electrical resistance (TER) demonstrated that gentamicin and 1% DMSO did not affect IPEC-J2 cell monolayer integrity via the disruption of cell membranes. Chromatographic data also ascertained that gentamicin penetration across the cell monolayer even in the presence of 1% DMSO was negligible at 6 h after the beginning of apical gentamicin administration. This study further indicates that the addition of this organic solvent does not increase the incidence of toxic effects related to gentamicin permeation

Strength of thermal undulations of phospholipid membranes

The temperature dependence of intermembrane interactions in freely suspended multilamellar membranes of dimiristoylphosphatidylcholine in D2O was studied using small-angle neutron scattering (SANS) and high-resolution x-ray diffraction (HRXRD). The study reveals that the Helfrich's undulation force is the dominating repulsion force at temperatures above 48.6 degrees C and intermembrane distances larger than 20.5 A. At similar to 77 degrees C the onset of the unbinding transition in the multilamellar membranes is observed. This transition has a continuous behavior in agreement with theoretical predictions and proceeds in accordance with a two-state model. Complimentary analysis of SANS and HRXRD data permits accurate calculation of the fundamental undulation force constant c(fl). The obtained value of c(fl)=0.111 +/- 0.005 is in good agreement with theoretical calculations. The results of this work demonstrate a key role of Helfrich's undulations in the balance of intermembrane interactions of lipid membranes under physiological temperatures and suggest that thermal undulations play an important part in the interactions of biological membranes. The agreement of the predictions with the experimental data confirms that lipid membranes can be considered as random fluctuating surfaces that can be described well by current theoretical models and that they can serve as a powerful tool for studying behavior of such surfaces