Colloidal particles for the delivery of steroid glycosides.

Water insoluble bioactive molecules with very high melting temperature and low solubility in water are difficult to formulate in food products. We demonstrate the synthesis of nanoscale particles from steroid glycosides using a facile liquid antisolvent precipitation method in the presence of various food grade stabilizers. Colloidal particles with sizes well below 200 nm are prepared from steroid glycosides containing extracts, as well as mixtures with phytosterol. In the mixtures, the formation of the typical for the phytosterol rod-like particles is suppressed. Particle size and structure are investigated by electron microscopy and dynamic light scattering. Due to the presence of surface charge and steric stabilization, colloidal particles do not display aggregation and are stable for a period of longer than one year. The results of this study are important for the formulation and delivery of steroid glycoside and phytosterol bioactive molecules in the fields of food, nutraceuticals, and medical applications.

Label-free imaging of biomolecules in food products using stimulated Raman microscopy.

The development of methods that allow microscale studies of complex biomaterials based on their molecular composition is of great interest to a wide range of research fields. We show that stimulated Raman scattering (SRS) microscopy is an excellent analytical tool to study distributions of different biomolecules in multiphasic systems. SRS combines the label-free molecular specificity of vibrational spectroscopy with an enhanced sensitivity due to coherent excitation of molecular vibrations. Compared to previous imaging studies using coherent anti-Stokes Raman scattering microscopy, the main advantage of SRS microscopy is the absence of the unwanted nonresonant background, which translates into a superior sensitivity and undistorted vibrational spectra. We compare spectra of complex materials obtained with stimulated Raman scattering and spontaneous Raman scattering in the crowded fingerprint region.We find that, as expected, there is excellent correspondence and that the SRS spectra are free from interference from background fluorescence. In addition, we show high-resolution imaging of the distributions of selected biomolecules, such as lipids and proteins, in food products with SRS microscopy.

Label-free imaging of biomolecules in food products using stimulated Raman microscopy.

The development of methods that allow microscale studies of complex biomaterials based on their molecular composition is of great interest to a wide range of research fields. We show that stimulated Raman scattering (SRS) microscopy is an excellent analytical tool to study distributions of different biomolecules in multiphasic systems. SRS combines the label-free molecular specificity of vibrational spectroscopy with an enhanced sensitivity due to coherent excitation of molecular vibrations. Compared to previous imaging studies using coherent anti-Stokes Raman scattering microscopy, the main advantage of SRS microscopy is the absence of the unwanted nonresonant background, which translates into a superior sensitivity and undistorted vibrational spectra. We compare spectra of complex materials obtained with stimulated Raman scattering and spontaneous Raman scattering in the crowded fingerprint region. We find that, as expected, there is excellent correspondence and that the SRS spectra are free from interference from background fluorescence. In addition, we show high-resolution imaging of the distributions of selected biomolecules, such as lipids and proteins, in food products with SRS microscopy.

The cochlear targets of cisplatin: an electrophysiological and morphological time-sequence study.

Cisplatin ototoxicity has at least three major targets in the cochlea: the stria vascularis, the organ of Corti, and the spiral ganglion. This study aims to differentiate between these three targets. In particular, we address the question of whether the effects at the level of the organ of Corti and spiral ganglion are mutually dependent or whether they develop in parallel. This question was approached by studying the ototoxic effects while they develop electrophysiologically and comparing these to earlier presented histological data [Van Ruijven et al., 2004. Hear. Res. 197, 44-54]. Guinea pigs were treated with intraperitoneal injections of cisplatin at a dose of 2 mg/kg/day for either 4, 6, or 8 consecutive days. This time sequence has not revealed any evidence of one ototoxic process triggering another. Therefore, we have to stay with the conclusion of Van Ruijven et al. (2004) that both processes run in parallel.

Immunohistochemical detection of platinated DNA in the cochlea of cisplatin-treated guinea pigs.

Cisplatin-induced ototoxicity is correlated with functional and morphological changes in the organ of Corti, the stria vascularis and the spiral ganglion. However, the cochlear sites of cisplatin uptake and accumulation have not been properly identified. Therefore, we have developed an immunohistochemical method to, indirectly, detect cisplatin in semithin cryosections of the guinea pig cochlea (basal turn) using an antiserum containing antibodies against cisplatin-DNA adducts. Platinated DNA was present in the nuclei of most cells in the organ of Corti and the lateral wall after cisplatin administration. Nuclear immunostaining was most pronounced in the outer hair cells, the marginal cells and the spiral ligament fibrocytes. This study is the first to demonstrate the presence of cisplatin in histological sections of the cochlea.

Time sequence of degeneration pattern in the guinea pig cochlea during cisplatin administration. A quantitative histological study.

We investigated the key tissues that are implicated in cisplatin ototoxicity within the time window during which degeneration starts. Guinea pigs were treated with cisplatin at a dose of 2 mg/kg/day for either 4, 6, or 8 consecutive days. Histological changes in the organ of Corti, the stria vascularis and the spiral ganglion were quantified at the light microscopical level. Outer hair cell (OHC) loss started between 4 and 6 days of cisplatin administration, but is only significantly different from the non-treated group after 8 days of treatment. Midmodiolar OHC counts were comparable to the cytocochleogram data. The cross-sectional area of the stria vascularis did not differ from the non-treated group, nor did an endolymphatic hydrops develop during the course of treatment. Spiral ganglion cell (SGC) densities did not decrease. After 6 days, however, detachment of the myelin sheath of the type-I SGCs was seen in the lower basal turn, whereas after 8 days it was also present in the more apically located turns. Myelin sheath detachment is the result of perikaryal shrinkage and swelling of the myelin sheath. The present study confirms that cisplatin at a daily dose of 2 mg/kg has a detrimental effect on the OHCs as well as on the type-I SGCs. These intracochlear effects occur simultaneously; OHC loss and SGC shrinkage start between the fourth and sixth day of cisplatin administration and appear to develop in parallel. At this dose, no histological effect on the stria vascularis could be observed, although previous electrophysiological experiments demonstrated a clear effect on the endocochlear potential