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L M Crowe

Publications and source records attributed to L M Crowe.

69 records · Page 4Linked to original sources
PubMed1985

Degradation of functional integrity during long-term storage of a freeze-dried biological membrane.

Trehalose, and to some extent a few other carbohydrates, is capable of stabilizing the structure and function of isolated biological membranes during lyophilization. In this paper the results of investigations into the long-term stability of the lyophilized membrane-carbohydrate mixtures were reported. The effects of varying water content, oxygen level, and light on the rates of oxidation, browning, and degradation of biological activity were reported. The efficiency with which three carbohydrates stabilized membrane structure was also reported, with glucose shown to be less efficient than maltose or trehalose. Increased water content accelerated loss of biological activity, possibly because, under the same conditions, nonenzymatic browning and photooxidation were accelerated also. Glucose-containing samples were especially unstable at elevated humidities. Efficiency of preservation could be maximized by storage under conditions of low oxygen, low humidity, and dark, and by the inclusion of high levels of trehalose.

Animals↗
PubMed1984

Preservation of functional integrity during long term storage of a biological membrane.

Sarcoplasmic reticulum vesicles freeze-dried in the presence of trehalose retain most of their original biological activity for short periods. When the dry vesicles are stored for longer periods in air, Ca2+-transport becomes uncoupled from ATPase activity within a few days. However, when they are stored under vacuum, ATPase activity, Ca2+ transport, and coupling between Ca2+ transport and ATP utilization are maintained essentially intact for at least 110 days.

Adenosine Triphosphatases↗
PubMed1984

Effects of carbohydrates on membrane stability at low water activities.

The relative effectiveness of a variety of carbohydrates in preserving the structural and functional integrity of membranes at low water activities was studied, using Ca-transporting microsomes from muscle as a model membrane. The order of effectiveness (greatest to lowest) was: trehalose, lactose, maltose, cellobiose, sucrose, glucose, fructose, sorbitol, raffinose, myo-inositol, glycerol. At the highest concentrations of the most effective sugars tested, microsomes were obtained upon rehydration that were similar structurally and functionally to fresh membranes. The least effective carbohydrates, alcohol sugars, all appear to be fusogenic. A structural explanation for relative effectiveness of the sugars was sought, but no clear relationship was found, except that effectiveness does not appear to be related to the number of position of hydroxyl groups available for hydrogen bonding.

Animals↗
PubMed1984

Interactions of phospholipid monolayers with carbohydrates.

Surface pressure studies of phospholipid monomolecular films of dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) formed at an air/water interface have been made and the effects on the films studied when various carbohydrates are present in the subphase. The results obtained show that at a given temperature, the area per molecule of DPPC increases with increasing concentration of the carbohydrate in the subphase. The carbohydrate which has the greatest expanding effect on the phospholipid monolayer is glycerol, followed in turn by trehalose, sucrose, glucose, raffinose, and inositol. The mechanism of monolayer expansion by glycerol is different from that observed in other carbohydrates, as the following experiments demonstrate. Below the phase transition temperature of DPPC, the area per molecule of DPPC at a pressure of 12.5 dyn/cm is the same with and without glycerol in the subphase. However, when the monolayer is heated to a temperature above the phase transition temperature for DPPC, the area/molecule on glycerol is considerably greater than the area/molecule on water at the same surface pressure. Cooling the monolayer back to the lower temperature produces an area/molecule of DPPC which is identical on both water and glycerol subphases. Glycerol therefore has no effect on the low-temperature (condensed) monolayers but causes expansion of the high-temperature (expanded) monolayers. By contrast with glycerol, both trehalose and sucrose interact with the DPPC monolayer producing an increased area/molecule over that observed on water, both with low-temperature (condensed) monolayers and with the high-temperature (expanded) monolayers. The efficiency of these carbohydrates at expanding the monolayer films (with the exception of glycerol) shows a strong correlation with their ability to stabilize membrane structure and function at low water contents.

Carbohydrates↗
PubMed1984

Infrared spectroscopic studies on interactions of water and carbohydrates with a biological membrane.

Infrared spectroscopy was used to investigate the changes in bands assigned to phospholipids and proteins in dehydrated and rehydrated sarcoplasmic reticulum. The changes in CH2 and CH3 stretching bands, amide bands, and phosphate stretching bands are similar to shifts in frequency seen for those bands in phospholipid and protein preparations during thermotropic phase transitions and hydration. IR studies on dry trehalose-sarcoplasmic reticulum mixtures show similar results; with increasing trehalose concentration in the dry mixtures, amide and phosphate bands shift to frequencies characteristic of hydrated samples. Changes in bands assigned to OH deformations in the trehalose suggest that the interaction between the carbohydrate and membrane is by means of hydrogen bonding between these -OH groups and membrane components.

Animals↗
PubMed1983

Preservation of structural and functional activity in lyophilized sarcoplasmic reticulum.

Ca-transporting microsomes isolated from abdominal muscle of lobster were lyophilized in the presence and absence of trehalose. The dry membranes appeared with freeze fracture to collapse into cup-shaped structures embedded in a matrix of trehalose when lyophilized in the presence of trehalose. Upon rehydration, the dry membranes yielded vesicles that were morphologically indistinguishable from freshly prepared ones. These rehydrated vesicles also showed ATPase activity and Ca transport only slightly different from those activities in freshly prepared vesicles. The concentration of trehalose required to achieve this degree of stabilization is about 0.3 g trehalose/g membrane. When the membranes were dried at lower trehalose concentrations extensive fusion occurred between vesicles, along with lateral phase separations of membrane proteins and lipids. The rehydrated vesicles showed poor Ca uptake and coupling between ATPase activity and Ca uptake. The membranes may also be stabilized in the dry state by lyophilizing them in sucrose, but about three times as much sucrose is required as trehalose.

Animals↗
PubMed1981

Activation of the contractile system in crustacean muscle: ultrastructural evidence for the role of the T system.

Freeze-fracture and thin sections of lobster abdominal fast flexor muscle were used to study the morphology of the sarcoplasmic reticulum (SR) and T system of crustacean muscle. Tannic acid mordanting, which can result in a dense black deposit in the T system lumen, was used to distinguish T system from SR membranes. Ferritin was also used as an extracellular tracer to confirm the tannic acid method. The T system consists of an extensive network of flattened sacs which fills most of the space between the myofibrils and is in close contact with them. The SR also appears as flattened sacs, sometimes with fenestrations. There is extensive junctional contact between the SR and T system. Quantitative estimates of the volume and surface area of the membranes show that the T system has about 50% more surface area than the SR. The intramembrane particle (IMP) density of the PF face of the T system is about 1100/micron 2 membrane, while the IMP density of the PF face of SR is about 4800/micron 2 membrane. In morphology, extent, and IMP density, the T system of lobster abdominal fast flexor muscle appears (AFF) adapted to provide at least part of the Ca2+ for muscle activation and the transport system for relaxation.

Animals↗
PubMed1979

Stereologic analysis of dystrophic chicken muscle.

Stereologic methods have been used to estimate the volume and surface densities of sarcoplasmic reticulum (SR) and T tubules of normal and dystrophic chicken pectoralis muscle fibers. The surface and volume densities of the T system in dystrophic muscle fibers showed large increases compared with normal muscle fibers; the surface and volume densities of the SR showed large decreases. In addition, the SR and T system in dystrophic fibers undergo changes in shape. The tubules of the free SR become much narrower; the T system becomes dilated and vesiculated. Dystrophic fibers, on the average, are much larger than normal but maintain the same sarcolemmal surface/fiber volume ratio as normal fibers. Alterations in the surface and volume densities of the dystrophic sarcotubular system may account for some of the altered contractile properties of these muscles.

Animals↗
PubMedDate not supplied

Physical properties of membrane fractions isolated from human platelets: implications for chilling induced platelet activation.

In previous studies, it has been suggested that chilling induced activation of human platelets is related to a lipid phase transition seen in membrane lipids. Those studies showed a single, surprisingly cooperative transition in human platelets, as determined by Fourier transform infrared (FTIR) spectroscopy, findings that are confirmed here with calorimetric measurements. Such transitions have now been studied in membrane fractions obtained from the platelets and it is reported that all fractions and purified phospholipids show similar transitions. In order to obtain these data it was necessary to develop means for separating these fractions. Therefore, a novel method for isolation and separation of dense tubular system (DTS) and plasma membranes in human platelets is described here. Lipid analysis showed that phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were the dominant phospholipids in both fractions, whereas cholesterol and sphingomyelin (SM) were predominantly located in the plasma membranes. Thermotropic phase transitions in the two membrane fractions, determined by differential scanning calorimetry (DSC) and FTIR spectroscopy were found to occur at about 15 degrees C, similar to the Tm of intact human platelets. These data are discussed in relation to the role of the DTS and plasma membranes in the cold-induced activation of human platelets.

Blood Platelets↗