Satoru Naito

Protein Secondary Structure Imaging with Ultrabroadband Multiplex Coherent Anti-Stokes Raman Scattering (CARS) Microspectroscopy

Protein secondary structures in human hair have been studied with ultrabroadband multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopy. The CARS peak-shift mapping method has been developed and applied to hair samples with and without treatments by chemical reduction and mechanical extension. It clearly visualizes the treatment induced changes in protein secondary structures and their spatial distributions. Using the new imaging technique, we found a multilayered structure in the human hair cortex.

In vivo measurement of human dermis by 1064 nm-excited fiber Raman spectroscopy

Background/aims: Although chemical information on the dermis in vivo is highly important in skin research, an efficient method for gathering this information is yet to be developed. Here, we demonstrate that newly developed near‐infrared (1064 nm) excited Raman spectroscopy is a powerful method for chemical analysis of human skin in vivo.

MICROWAVE DIELECTRIC ANALYSIS OF HUMAN STRATUM CORNEUM IN VIVO

The dielectric properties of the human skin stratum corneum (SC) in the frequency range higher than 107 Hz are not well understood because of the difficulty in selective scanning of the SC area in vivo. The present study was carried out to make clear factors responsible for the dielectric properties using a measuring system specially developed for the study of SC [S. Naito, M. Hoshi, S. Mashimo, Anal. Biochem. 251 (1997) 163-172]. We found that the dielectric properties of SC can be expressed by the linear combination of two relaxation processes and d.c. conduction. The faster relaxation is that of free water. The slower relaxation and d. c. conduction were analyzed using a model assuming interfacial polarization between dissimilar materials. We concluded that the polarization is the origin of the slower relaxation process because the experimental data could be well interpreted according to the above mechanism. We also concluded that the polarization of swelled SC locates at the interface between SC cells and the intercellular lipid layer, or at the interface between the lipophilic and the hydrophilic part of the lamellar structured intercellular lipid layer.

Microwave Dielectric Study on Water Structure and Physical Properties of Aqueous Systems Using Time Domain Reflectometry with Flat-End Cells

Microwave dielectric measurements were performed in the frequency range from 1 mHz up to 30 GHz using a time domain reflectometry (TDR) method for emulsions and gels. Flat-end sample cells have been used in the TDR measurement to contact a small spot of the surface of those viscoelastic and solid samples without any destruction. Relaxation processes due to various water structures were observed for these aqueous systems. Relaxation parameters thus obtained offer information about these water structures and amounts. The relaxation strength obtained from the high frequency process due to free water can be an adequate measure of water content in spite of some ambiguities for different water structures in some materials. Comparisons of actual water contents in emulsion with those estimated from the relaxation strength indicate that water structure is affected by the interaction between water and micelle. Unfreezable water observed in DNA gel under the freezing point consists of bound water and a fraction of free water. Bound water molecules are still unfreezable to keep the double helical structure of DNA, when the fraction of free water is frozen at lower temperatures. These water structures determine physical properties of moist materials. TDR measuring technique with the flat-end cell is effective to investigate water structures in viscoelastic moist materials and to evaluate physical properties and structures of complex molecular systems.

Hydrating effect of potassium lactate is caused by increasing the interaction between water molecules and the serine residue of the stratum corneum protein

Abstract:  Natural moisturizing factors (NMFs) play an important role in maintaining the physical properties of the stratum corneum (SC). The relationship between SC water content and NMFs has long been investigated. Recently, we demonstrated that potassium lactate as an NMF increased SC water content more than sodium lactate did. The details of the moisturizing mechanism of NMFs, however, were not revealed. We, therefore, investigated the cause of the SC moisturizing effect of potassium lactate in comparison with sodium lactate. Using differential scanning calorimetry, we found that potassium lactate increased the bound water content of plantar SC more than what sodium lactate did. We also found, however, that the bound water content of the potassium lactate solution was less than that of the sodium lactate solution, suggesting that potassium lactate increased the water molecules interacting with SC components. Moreover, potassium lactate increased the ratio of hydrogen/deuterium exchange at 1340/cm, which represents the OH bending mode, of plantar SC spectra obtained by the attenuated total reflectance infrared spectroscopy. We assign this band to the OH group of the serine residue. These results suggest that potassium lactate increases the water‐holding capacity of the SC by increasing interaction between water molecules and the OH group of serine in SC keratin.

A method of measuring surface permittivity by microwave dielectric analysis

A method for controlling skin depth by microwave dielectric analysis using an open end cell is presented here. The correlation between the measured permittivity, the electric length of the cell, and the permittivity depth profile has been shown from time domain reflectometry measurements on a sample with known depth profile of permittivity. This skin depth controlling method is exceedingly useful for dielectric analysis of water in living organisms and related food products.

A study of the behavior and mechanism of thermal conduction in the skin under moist and dry heat conditions

We analyzed skin heat conduction under moist and dry heat conditions to confirm the influence of moist heat on the skin and subcutaneous region.

Surfactant Uptake Dynamics in Mammalian Cells Elucidated with Quantitative Coherent Anti-Stokes Raman Scattering Microspectroscopy

The mechanism of surfactant-induced cell lysis has been studied with quantitative coherent anti-Stokes Raman scattering (CARS) microspectroscopy. The dynamics of surfactant molecules as well as intracellular biomolecules in living Chinese Hamster Lung (CHL) cells has been examined for a low surfactant concentration (0.01 w%). By using an isotope labeled surfactant having CD bonds, surfactant uptake dynamics in living cells has been traced in detail. The simultaneous CARS imaging of the cell itself and the internalized surfactant has shown that the surfactant molecules is first accumulated inside a CHL cell followed by a sudden leak of cytosolic components such as proteins to the outside of the cell. This finding indicates that surfactant uptake occurs prior to the cell lysis, contrary to what has been believed: surface adsorption of surfactant molecules has been thought to occur first with subsequent disruption of cell membranes. Quantitative CARS microspectroscopy enables us to determine the molecular concentration of the surfactant molecules accumulated in a cell. We have also investigated the effect of a drug, nocodazole, on the surfactant uptake dynamics. As a result of the inhibition of tubulin polymerization by nocodazole, the surfactant uptake rate is significantly lowered. This fact suggests that intracellular membrane trafficking contributes to the surfactant uptake mechanism.

In vivo evaluation of lateral lipid chain packing in human stratum corneum

The matrix of intercellular lipids (ICL) of stratum corneum (SC) plays an important role in the barrier function of SC. It is important to understand the structure of the ICL matrix for dermatology and cosmetic science. Several methods exist for the analysis of the structure; however, it is difficult to conduct these analyses noninvasively.