In Vitro and In Vivo Evaluation of Photocontrolled Biologically Active Compounds - Potential Drug Candidates for Cancer Photopharmacology.

Photocontrolled, biologically active compounds are an emerging class of "smart" drug candidates. They provide additional safety in systemic chemotherapy due to their precise spatiotemporal activation by directing a benign, non-ionizable light to a specific location within the patient's body. This paper presents a set of methods to evaluate the in vitro potency and ex vivo efficiency of the photoactivation of photocontrolled, biologically active compounds as well as the in vivo efficacy at early stages of drug development. The methodology is applied to anticancer cytotoxic peptides, namely, the diarylethene-containing analogs of a known antibiotic, gramicidin S. The experiments are performed using 2D (adherent cells) and 3D (spheroids) cell cultures of a cancer cell line (Lewis lung carcinoma, LLC), live tissue surrogates (pork meat mince), and an allograft cancer model (subcutaneous LLC) in immunocompetent mice. The selection of the most effective compounds and estimation of realistic phototherapeutic windows are performed via automated fluorescence microscopy. The photoactivation efficiency at varying illumination regimens is determined at different depths in a model tissue, and the optimal light dosage is applied in the final therapeutic in vivo experiment.

Adaptive wavefront sensor based on the Talbot phenomenon.

A new adaptive method of wavefront sensing is proposed and demonstrated. The method is based on the Talbot self-imaging effect, which is observed in an illuminating light beam with strong second-order aberration. Compensation of defocus and astigmatism is achieved with an appropriate choice of size of the rectangular unit cell of the diffraction grating, which is performed iteratively. A liquid-crystal spatial light modulator is used for this purpose. Self-imaging of rectangular grating in the astigmatic light beam is demonstrated experimentally. High-order aberrations are detected with respect to the compensated second-order aberration. The comparative results of wavefront sensing with a Shack-Hartmann sensor and the proposed sensor are adduced.

Bottlenecks of the wavefront sensor based on the Talbot effect.

Physical constraints and peculiarities of the wavefront sensing technique, based on the Talbot effect, are discussed. The limitation on the curvature of the measurable wavefront is derived. The requirements to the Fourier spectrum of the periodic mask are formulated. Two kinds of masks are studied for their performance in the wavefront sensor. It is shown that the boundary part of the mask aperture does not contribute to the initial data for wavefront restoration. It is verified by experiment and computer simulation that the performance of the Talbot sensor, which meets established conditions, is similar to that of the Shack-Hartmann sensor.