Cell-selective intracellular targeting is a key element of more specific and safe enzyme, toxin, and gene therapies. Endothelium poorly internalizes certain candidate carriers for vascular immunotargeting, such as antibodies to platelet endothelial cell adhesion molecule 1 (PECAM-1). Conjugation of poorly internalizable antibodies with streptavidin (SA) facilitates the intracellular uptake. Although both small and large (100-nm versus 1000-nm diameter) anti-PECAM/SA-beta galactosidase (SA-beta-gal) conjugates bound selectively to PECAM-expressing cells, only small conjugates showed intracellular accumulation of active P-gal. To study whether size of the conjugates controls the uptake, a series of anti-PECAM/SA and anti-PECAM/bead conjugates ranging from 80 nm to 5 mum in diameter were produced. Human umbilical vein endothelial cells and PECAM-transfected mesothelioma cells internalized 80- to 350-nm anti-PECAM conjugates, but not conjugates larger than 500 nm. Further, size controls intracellular targeting of active therapeutic cargoes in vitro and in vivo. Small anti-PECAM/DNA conjugates transfected target cells in culture 5-fold more effectively than their large counterpart (350- versus 4200-nm diameter). To evaluate the practical significance of the size-controlled subcellular addressing, we coupled glucose oxidase (GOX) to anti-PECAM and antithrombomodulin. Both types of conjugates had equally high pulmonary uptake after intravenous injection in mice, yet only small (200- to 250-nm), not large (600- to 700-nm), GOX conjugates caused profound oxidative vascular injury in the lungs, presumably owing to intracellular generation of H2O2. Thus, engineering of affinity carriers of specific size permits intracellular delivery of active cargoes to endothelium in vitro and in vivo, a paradigm useful for the targeting of drugs, genes, and toxins. (C) 2002 by The American Society of Hematology.

A new type of hybrid material is prepared through sol-gel processing by the polycondensation of (MeO)(3)Si-R-Si(OMe)(3) units containing a rigid organic anisotropic group R. The hybrid covalently bounded organic-inorganic gel shows an unexpected structural birefringence Deltan(Deltan = 2 x 10(-3)). This birefringence is induced by a strain field anisotropy during gel aging as revealed by simultaneous dynamic light scattering and birefringence measurements. To better address the role of the strain field, we use a free interface sol-gel/air to control strain anisotropy and to measure the gel optical axis and its birefringence. We find that the birefringence is associated to a long-range orientational order of organic moieties induced by the strain anisotropy during the gel aging.

Velocity fluctuations in sedimentation are studied to investigate the origin of a hypothesized universal scale [P. N. Segre, E. Herbolzheimer, and P. M. Chaikin, Phys. Rev. Lett. 79, 2574 (1997)]. Our experiments show that fluctuations decay continuously in time for sufficiently thick cells, never reaching steady state. Simulations and scaling arguments suggest that the decay arises from increasing vertical stratification of particle concentration due to spreading of the sediment front. The results suggest that the velocity fluctuations in sedimentation depend sensitively on cell geometry.