Cytofectins are positively charged lipophilic molecules that readily form complexes with DNA and other anionic polynucleotides. Normally, cytofectins are combined with an activity-augmenting phospholipid such as dioleoylphosphatidylethanolamine (DOPE), and a film of dried, mixed lipid is prepared and hydrated to form cationic liposomes. The liposome solution is then mixed with a plasmid DNA solution to afford cytofectin-DNA complexes which, when presented to living cells, are internalized and the transgene is expressed. One of the most potent cytofectins, dimyristoyl Rosenthal inhibitor ether (DMRIE), is presently being used to deliver transcriptionally active DNA into human tumor tissues. Here we report the remarkable consequences of replacing the alcohol moiety of DMRIE with a primary amine. The resulting cytofectin, called beta-aminoethyl-DMRIE (betaAE-DMRIE), promoted high level transfection over a broad range of DNA and cationic lipid concentrations. A comparison of in vitro transfection activity between DMRIE and betaAE-DMRIE in 10 cell types revealed that betaAE-DMRIE was more active than DMRIE, and that betaAE-DMRIE, unlike DMRIE, was maximally effective in the absence of colipid. The consequences of the alcohol-to-amine conversion on the structure of the cytofectin/DNA complex was also examined by Atomic Force Microscopy. Strikingly dissimilar images were found for plasmid DNA alone and for the plasmid complexes of betaAE-DMRIE and DMRIE/DOPE.

Summary:We have investigated the effects of tip-sample forces and relative humidity when using a scanning force microscope (SFM) to image DNA molecules adsorbed on fresh mica. As the force between the tip and the sample increases, the apparent height of the DNA molecules decreases. After being imaged with high forces, the DNA molecules recover partially in their apparent height, indicating that a plastic deformation of the DNA has been induced by the scanning tip. At low humidities, DNA molecules can be imaged with a force up to 150 nN during the scanning without obvious damages. At higher humidities, however, the DNA molecules can be dissected or swept away by the tip even at a tip-sample force of 30 nN. The net force between the tip and the molecules is the vector sum of several forces, the dominant components of which are the elastic force due to the cantilever bending and the capillary force resulting from the water meniscus formed between the tip and the sample surface. When the relative humidity of the imaging environment is increased, the capillary force becomes stronger.

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A fully automated experimental setup is described that allows the determination of the heat capacity and thermal diffusivity on the same piece of a nonconducting thin solid sample. Both properties are measured using ac techniques. Measurements have been made on microscope glass cover slides for temperatures between 25 and 300 K. The values obtained for the specific heat are within 5% of those reported in the literature and the values of the thermal diffusivity are within 10% of reported values.