Plant fructans stabilize phosphatidylcholine liposomes during freeze-drying.

Fructans have been implicated as protective agents in the drought and freezing tolerance of many plant species. A direct proof of their ability to stabilize biological structures under stress conditions, however, is still lacking. Here we show that inulins (linear fructose polymers) isolated from chicory roots and dahlia tubers stabilize egg phosphatidylcholine large unilamellar vesicles during freeze-drying, while another polysaccharide, hydroxyethyl starch, was completely ineffective. Liposome stability was assessed after rehydration by measuring retention of the soluble fluorescent dye carboxyfluorescein and bilayer fusion. Inulin was an especially effective stabilizer in combination with glucose. Analysis by HPLC showed that the commercial inulin preparations used in our study contained no low molecular mass sugars that could be responsible for the observed stabilizing effect of the fructans. Fourier transform infrared spectroscopy showed a reduction of the gel to liquid-crystalline phase transition temperature of dry egg PtdCho by more than 20 degrees C in the presence of inulin. A direct interaction of inulin with the phospholipid in the dry state was also indicated by dramatic differences in the phosphate asymmetric stretch region of the infrared spectrum between samples with and without the polysaccharide.     

Trehalose and dry dipalmitoylphosphatidylcholine revisited

Dry mixtures of sonicated vesicles of DPPC and trehalose which contained a maximum of 0.2 mol water/mol lipid were examined by differential scanning calorimetry, Fourier transform infrared spectroscopy and freeze-fracture electron microscopy. Samples of dry DPPC and trehalose prepared from aqueous solution had a minimum T_m of 24°C for the gel to liquid-crystalline transition provided that the vesicles were dried with trehalose while the lipid was in liquid-crystalline phase. This low transition is compared to a transition of 105–112°C for dry pure DPPC and of 42°C for hydrated pure DPPC. The present work is an extension of earlier work from this laboratory using both other lipids and other methods of preparation.     

The response of chelonian muscle spindles to mechanical stimulation

Single unit recordings have been made from the muscle spindles of the extensor digitorum brevis I muscle of the chelonian emys orbicularis. The responses to ramp-type mechanical stretches (up to 5 mm s^?1 velocity and up to 1.6 mm extension) were compared to those of spindles from other groups. It is found that the spindles have lower rates of firing than those from the other groups with the exception of the snake spindles. Generally the spindles behaved like the secondary ending of mammalian spindles or the tonic type of snake spindle in terms of their response to the velocity of stretch. The results are consistent with the view that the tonic response arises from intrafusal muscle fibres in which the sensory region has a structure which is fairly uniform and similar to that of the polar regions and not interrupted by accumulations of nuclei.     

Inhibition of dehydration-induced fusion between liposomal membranes by carbohydrates as measured by fluorescence energy transfer

The relative abilities of a number of naturally occurring carbohydrates to inhibit dehydration-induced fusion between palmitoyloleoylphosphatidylcholine:phosphatidylserine (85:15) large unilamellar vesicles have been studied. Fusion events were quantified using a fluorescence resonance energy transfer technique. Trehalose was most effective at inhibiting fusion (0.4 g/trehalose/g lipid showed 30% probe intermixing), followed by maltose (60% intermixing), fructose (60%), sucrose (70%), glucose (80%), cellobiose, glycerol, raffinose, and myo-inositol (90%). The relative abilities of these carbohydrates to inhibit fusion correlate directly with their abilities to interact with phospholipids, maintain bilayer fluidity, and preserve biological membranes. The results are discussed in relation to the crystalline structure of the carbohydrates and their possible influence on level of interaction with phosphate head groups.