Influence of Temperature on Stability of Multilamellar Liposomes in Wool Dyeing

Liposomes are lipid vesicles that are composed of amphiphile molecules and can carry hydrophobic and hydrophilic materials. In this research work liposomes used as carrier for transfer of dye molecules into wool fibers. The preparation and production of multilamellar liposomes (MLV) from Soya lecithin were carried out and the behavior of liposomes at different temperature was studied. The effect of different concentration of liposomes in the dye exhaustion profile of two dyes (Namely, Irgalan Blue FBL and Lanaset Blue 2R) at two different temperatures of 85°C and 95°C on the wool fabric was investigated. The results showed that presence of liposomes in the dye-bath helps to increase the dye absorption on the wool fabric before 80°C. Dyeing at higher temperature and longer time leads to a decrease in the final exhaustion along with increase in the liposomes concentration. Liposomes at high temperature converted to the disperse phospholipids unimers that may deposited on the fabric surface and may produce a hydrophobic barrier against absorption of dye. The presence of 1% o.w.f. (on weight of fabric) of liposomes at 85°C improved the dye exhaustion of Irgalan Blue FBL on the wool fabric. The wash fastness properties of samples which dyed in the dye-bath containing liposomes also improved.     

Spectral changes in membrane fragments and artificial liposomes of Anacystis induced by chilling.

Isolated membrane fragments from Anacystis nidulans grown at 39 °C undergo visible spectral changes on chilling, suggesting a carotenoid component is altered. No such changes are seen when cells are grown at 25 °C. The magnitude of the decreased absorbance is a function of the chilling temperature and the media in which membrane fragments are suspended. The spectral decrease following chilling develops relatively slowly and is a function of the cooling rate and final temperature. The absorbance change is reversed if the fragments are heated to near 50 °C subsequent to chilling. Liposomes prepared from a total lipid extract of Anacystis undergo a spectral change on chilling which closely resembles that occurring in whole cells or isolated membrane fragments. Liposomes prepared from an extract of cells grown at 25 °C show only about 30% as great a spectral change as those from cells grown at 39 °C. The spectral bleaching is freely reversible when the liposomes are reheated, but shows a pronounced hysteresis. It is suggested that specific phase changes occur in Anacystis membranes and artificial liposomes on cooling which alter the environment of carotenoid. These changes may relate to previous observations that cells grown at 39 °C cannot survive a cold shock while those grown at 25 °C do.     

Effect of streptolysin S on liposomes Influence of membrane lipid composition on toxin action

The effect of the bacterial cytolytic toxin, streptolysin S, on liposomes composed of various phospholipids was investigated. Large unilamellar vesicles containing [¹⁴C]sucrose were prepared by reverse-phase evaporation, and membrane damage produced by the toxin was measured by following the release of labeled marker. The net charge of the liposomes had little or no effect on their susceptibility to steptolysin S and the toxin was about equally effective on liposomes composed of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol. Experiments with liposomes composed of synthetic phospholipids showed that the ability of the toxin to produce membrane damage depended on the degree of unsaturation of the fatty acyl chains. The order of sensitivity was C18 : 2 phosphatidylcholine > C18 : 1 phosphatidylcholine > C18 : 0 phosphatidylcholine = C16 : 0 phosphatidylcholine. Liposomes containing the latter two phospholipids were virtually unaffected by streptolysin S, and experiments with C18 : 0 phosphatidylcholine suggested that toxin activity does not bind to liposomes composed of phospholipids with saturated fatty acyl chains. The inclusion of 40 mol% cholesterol in C16 : 0 phosphatidylcholine and C18 : 0 phosphatidylcholine liposomes made these vesicles sensitive to streptolysin S. Egg phosphatidylcholine liposomes, which were unaffected at 0°C and 4°C became susceptible to the toxin at these temperatures when cholesterol was included. Liposomes composed of C14 : 0 phosphatidylcholine were unaffected by streptolysin S at temperatures below the chain-melting transition temperature (23°C) of this phospholipid, but became increasingly susceptible above this temperature. The results suggest that the fluidity of the phospholipid hydrocarbon chains in the membrane is important in streptolysin S action.     

Characterization of liposomes prepared using a microemulsifier

A new type of device can prepare liposomes continuously, in large quantities and with excellent aqueous space capture efficiency. At initial lipid concentration of 300 μmol/ml these liposomes capture approx. 75% of cytosine arabinoside used as an aqueous space marker. Liposome size can be reduced by increasing the number of times the preparations are recycled through the microemulsifier. Liposomes less than 0.1 μm in diameter, as shown by electron microscopy, can be made easily. Liposomes prepared at 300 μmol/ml, composed of phosphatidylglycerol/phosphatidylcholine/cholesterol in a 0.1:0.4:0.5 molar ratio leaked less than 1% of entrapped cytosine arabinoside (Ara-C) at 4°C, and less than 10% Ara-C at 37°C plus serum, over a 48 h period. These liposomes could be useful for a number of applications including diagnostics, therapeutics and model membrane studies.     

Raman spectroscopic investigation of the interaction of gramicidin A with dipalmitoyl phosphatidylcholine liposomes

The interaction of gramicidin A with dipalmitoyl phosphatidylcholine liposomes is investigated by Laser-Raman spectroscopy. As revealed by the methylene CH stretching mode the phase transition of the hydrocarbon chains near 40°C is eliminated in the presence of gramicidin A. Liposomes prepared from a mixture of lecithin and cholesterol seem to be unaffected by gramicidin A and show only the normal broadened phase transition.     

Liposomes with Metronidazole for Topical Use: The Choice of Preparation Method and Vehicle

Abstract

Liposomes containing metronidazole were prepared for the treatment of skin disorder Rosacea. To optimize the composition and size of liposomes, natural and synthetic lipids were used in three different preparation methods. Optimal liposomal preparation was incorporated into five dermal vehicles (three O/W emulsion creams and two gels) and vehicles tested for the stability (at 20 and 40 °C) during a storage period of 4 weeks. The evaluation of the vehicles was based on the comparison between the original size distribution of liposomes and liposomes in vehicles (after 4 weeks) and on the rheological behaviour. The best vehicle appears to be the Carbopol? gel, in which the liposomal size stayed unchanged even at 40 °C during the 4 weeks period.     

Liposomes from the thermophilic eubacterium Thermus SP. fluorescence polarization and permeability properties

• 1.1. The physical properties of liposomes prepared from total and polar lipid extracts of Thermus SPS 11 and SPS 17 grown at 50°C and at the optimal growth temperature (73–75°C) have been studied using 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence polarization (P) and correlated to the permeability properties of the bilayers.
• 2.2. P data show the presence of a broad phase transition from a gel to a liquid-crystal phase for all liposomes studied. Liposomes from lipid fractions of Thermus SPS 11 and SPS 17 grown at 50°C were in a less fluid state than those of bacteria grown at the optimal growth temperature.
• 3.3. Comparing the permeabilities to ethyleneglycol, glycerol and erythritol of liposomes from lipid extracts of Thermus SPS 11 and SPS 17 grown at the two temperatures already mentioned, liposomes from lipid extracts of bacteria grown at 50°C were, in general, more permeable to ethyleneglycol and glycerol and less permeable to erythritol.
• 4.4. Liposomes from lipid fractions with higher carotenoid (CA) content were in a more rigid state within the whole range of temperatures studied, and their permeabilities to the three polyols were significantly lower than in the low-CA samples.

     

Resorcinolic lipids improve the properties of sphingomyelin–cholesterol liposomes

Alkylresorcinolic lipids isolated from cereal grains and their semi-synthetic myristoyl-sulphonyl derivatives (MSAR) were used to modify small long-circulating sphingomyelin–cholesterol liposomes. Those SM:Chol vesicles modified with 10–30 mol% resorcinolic lipids had stable size and low membrane permeability in vitro at 4 °C and 37 °C. Liposomes containing 30 mol% MSAR showed very fast solute release in the presence of human plasma at 37 °C, which was drastically diminished in heat-inactivated plasma. In vivo studies showed that unmodified SM:Chol liposomes and those modified with alkylresorcinols were eliminated from the circulation more slowly than liposomes with the highest concentration of MSAR in membrane and were located mostly in the liver and spleen.     

Thermolabile Liposomes with a High Fusion Efficacy at 42°C can be Made of Dipalmitoylphosphatidylcholine/Fatty Alcohol Mixtures in the Molar Ratio of 1/2

Abstract

Liposomes made of dipalmitoylphosphatidylcholine (DPPC²), dipalmitoyl-phosphatidylglycerol (DPPG), and different long-chain fatty alcohols were investigated with respect to their colloidal stability, chain-melting phase transition temperature, and temperature dependent inter-vesicle fusion. In particular, the practical usefulness of the stoichiometric 1/2 (mol/mol) mixtures of the phospholipids and fatty alcohols, mainly elaidoyl alcohol (EL-OH) were studied. The mole fraction of DPPG in the bilayers of such vesicles affects crucially the colloidal stability of the resulting lipid suspensions; at least 15 mol-% of DPPG (relative to DPPC) must be incorporated into the bilayers in order to make the liposome suspension colloidally sufficiently stable at room temperature. The corresponding DPPC/DPPG/EL-OH (0.85/0.15/2) mixed lipid vesicles undergo a lamellar-gel to inverted hexagonal (H_IT) phase transition at 52.7°C, however, and then fuse and aggregate massively. The related phase transition temperature of the DPPC/DPPG/palmitelaidoyl alcohol (0.85/0.15/2) mixture is 48.4°C. This indicates that the chain-melting phase transition temperature of the investigated lipid mixtures is rather sensitive to the alcohol chain-length. This transition temperature is independent, however, of the bulk proton concentration in the pH region between 4.9 and 7.2. Stoichiometric 1/2 mixtures of phospholipids and EL-OH have a high propensity for the inter-vesicle fusion at 42°C and neutral pH. The reason for such fusion 10°C below the lamellar-to-nonlamellar phase transition temperature are the defects that are generated during the chain-melting of the (partly segregated) phospholipid component at 42°C; the proximity of the lamellar to non-lamellar phase transition temperature of the phospholipid/fatty alcohol (1/2) complex at 52°C also plays an important role.     

The Design of a Pharmaceuttcally Acceptable Liposomal Formulation of Recombinant Interleukin-2 (Ril-2) for Locoregional Anticancer Immunotherapy

Abstract

Liposomes composed of egg-phosphatidylcholine and egg-phosphati-dylglycerol containing recombinant interleukin-2 (rIL-2) were prepared and characterised for use in locoregional anticancer immunotherapy. During the encapsulation studies it was observed that the protein precipitated. Therefore, the impact of the precipitation phenomenon on the characteristics and in vivo performance of rIL-2 liposomes was studied. Recombinant IL-2 was diluted in various aqueous media and the amount of precipitated protein determined. Also, the in vitro bioactivity, chemical stability, and in vivo antitumor efficacy of liposomes prepared with precipitated rIL-2 or non-precipitated rIL-2 were assessed.

Massive protein precipitation (60 up ± 95% of total protein) was observed upon dilution of rIL-2 in salt-containing media, but not upon dilution in 5% glucose or water. Liposomes prepared with precipitated rIL-2 were shown to release 50% of the entrapped rIL-2 over a three-day period at 37° in protein-containing media. Loss of rIL-2 bioactivity and chemical integrity was observed during storage at 4° over a 4-week period. Locoregional administration of precipitated rIL-2 in the guinea pig Line 10 tumor model resulted in significantly more tumor growth inhibition than administration of non-precipitated rIL-2. Liposomes containing non-precipitated rIL-2 were found to elicit similar antitumor effects as precipitated rIL-2. The results point to the importance of proper characterisation of new rIL-2 formulations as the physical properties of formulated rIL-2 may strongly influence its bioactivity.     

Stability of small unilamellar liposomes in serum and clearance from the circulation: The effect of the phospholipid and cholesterol components

Small unilamellar liposomes containing carboxyfluorescein (CF) and composed of various unsaturated and saturated phospholipids with or without cholesterol were incubated in the presence of mouse serum at 37°C. Liposomes composed of egg L-α-phosphatidylcholine (PC), L-α-dioleoylphosphatidylcholine (DOPC) or sphingomyelin (SM) became rapidly permeable to entrapped CF but incorporation of cholesterol into such liposomes reduced CF leakage. Under similar conditions, CF leakage from cholesterol-free liposomes composed of saturated phospholipids of increasing fatty acid chain length was dependant on the liquid-crystalline phase transition temperature (Tc) of the phospholipid component. Thus, L-α-dilaureoylphos-phatidylcholine (DLPC), L-α-dimyristoyl phosphatidylcholine (DMPC) and L-α-dipalmitoylphosphatidylcholine (DPPC) with Tc's below or near the temperature of the incubation (37°C) released CF rapidly whereas L-α-diheptedecanoyl phosphatidylcholine (DHPC), L-α-distearoylphosphatidylcholine (DSPC) and hydrogenated egg PC (HPC) liposomes with Tc's above 37°C retained the dye quantitatively. After incorporation of cholesterol into liposomes composed of saturated phospholipids, CF release was reduced for DLPC and DMPC and increased for DPPC, DSPC, DHPC and HPC vesicles. Liposomes with or without cholesterol exhibiting greatest stability (in terms of CF retention) in the presence of serum were injected intravenously into mice and rates of clearance of quenched CF from the circulation measured. Observed clearance rates were linear and, when liposomes contained tritiated phospholipid, identical to those of the radiolabel suggesting retention of liposomal integrity in the intravascular space. However, half-lifes of liposomes ranging from 0.1 to 16 h did not correlate with the physical characteristics of their phospholipid component. After intraperitoneal injection, there was quantitative entry of quenched CF (stable liposomes) into the blood from which it was eliminated at rates corresponding to those observed after intravenous injection. These results suggest that solute retention by liposomes and their half-life in the circulation can be controlled by the appropriate manipulation of liposomal membrane fluidity and composition.     

Effect of temperature on the development of the immature stages ofBembidion lampros [Coleoptera: Carabidae]

The effects of temperature on survival and development of immature stages ofBembidion lampros were examined under controlled conditions in the laboratory. The duration of development was examined at 5°C, 12°C, 17°C, 19°C, 22°C, 25°C, 30°C and 32°C and found to be inversely related to temperature. Between 12°C and 30°C there was a significant difference in duration of egg development, but no significant difference in percentage of eggs hatched. The upper and lower lethal limits for egg development were estimated (c 31°C and 4°C respectively). There are 3 larval instars and one pupal stage. Development of larva plus pupa required a minimum of 20.9 days at 30°C. Between 22°C and 30°C survival from larva to the adult stage was 80%.      

The development of Puccinia hordei on barley cv. Zephyr

Germination of uredospores of Puccinia hordei was similar on cover-slips and on the first leaves of barley seedlings (cv. Zephyr) at 100 % r.h. over the range 5–25 °C, being greatest at 20 °C. At 15, 20 and 25 °C maximum germination was attained in 6 h. No uredospores germinated on coverslips in humidities below saturation. The numbers of pustules which subsequently developed on plants incubated at 5, 10, 15 or 18 °C and 100 % r.h. for varying periods up to 24 h, were directly related to rise in temperature and length of incubation. The time from inoculation to eruption of pustules (generation time) was 6 days at 25 °C, 8 days at 20 °C, 10 days at 15 °C, 15 days at 10 °C and 60 days at 5 °C. Pustule production on inoculated plants which had been kept at 5 °C was rapidly accelerated when they were transferred to 20 °C. Data obtained at constant temperatures were used to predict generation times of the fungus in the field. The productivity of pustules, determined as weight of uredospores, was examined at 10, 15 and 20 °C. Significantly more spores were produced at 15 than at 10 °C and most were produced at 20 °C. The results are discussed in relation to those obtained by other workers and to the development of brown rust in the field.     

Interactive effects of photoperiod and temperature on diapause induction and termination in the yellow-spotted longicorn beetle, Psacothea hilaris

Abstract. The interactive effects of temperature (20 °C or 25 °C) and photoperiod (LD 12 : 12 h or LD 15 : 9 h) on diapause induction and termination are investigated in the west‐Japan type yellow‐spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae). Larval diapause of P. hilaris is induced under three diapause‐inducing conditions (20 °C–SD, 20 °C–LD and 25 °C–SD), and the diapause larvae are transferred to one of four conditions (20 °C–SD, 20 °C–LD, 25 °C–SD or 25 °C–LD) for observation of pupation, which indicates termination of diapause. The intensity of diapause induced under the three conditions increases in the order 20 °C–SD < 25 °C–SD < 20 °C–LD, when assessed by the time course of pupation after the transfer. On the other hand, the effectiveness of the temperature–photoperiod combinations to terminate diapause is in the order 25 °C–SD (ineffective) < < 20 °C–LD < 25 °C–LD < 20 °C–SD. Among the temperatures (5, 10, 15 and 20 °C) examined, 15 °C is the most effective in terminating diapause under the short day; diapause in most larvae appears to have been completed in 15 days.     

Effect of environmental temperature on development, fecundity, survival and predation of the snail-predatorSarcophaga misera (Dipt., Sarcophagidae)

Effect of temperature (10°–45°C) on development, fecundity, survival, population parameters, and rate of predation on snail by the flySarcophaga (Parasarcophaga) misera has been studied under controlled laboratory conditions. Temperatures ranging from 25°–30°C effected higher rate of development, fecundity, and predation on the snailIndoplanorbis exustus, than at 15°, 20°, 35° & 40°C. Longevity was maximum at 20°C. Implications of the role of temperature in influencing the biology and predation by this fly are discussed.     

The effect of starvation on size and survival of the terrestrial planarian Artioposthia triangulata (Dendy) (Tricladida: Terricola)

Ten Artioposthia triangulata were kept without food at 5°C, 10°C, 15°C, 20°C and 23°C. Survival and weights were recorded weekly. All planarians died within one week at 23°C and within three weeks at 20°C. One individual survived for 60 wk at 5°C. A negative exponential equation was fitted to average survival times for planarians kept at 5°C, 10°C, 15°C and 20°C. Negative exponential models could also describe the pattern of degrowth by planarians deprived of food. The results are discussed in relation to possible survival of the planarian outdoors in Northern Ireland and the implications for earthworm populations.     

Osmotic Dehydration of Liposomal Dispersions: Influence of Particle Size and Electrostatic Deposition of Cold Water Fish Skin Gelatin

Liposomes are a promising delivery system for bioactives in food and nutraceuticals. Their practical application is limited by their physical and chemical instability caused by extrinsic factors. The physical stability of liposomes of three different sizes coated with cold water fish skin gelatin was assessed during osmotic dehydration at 2, 21 and 70 °C. Soy lecithin was used to prepare 1 % liposomal dispersions. The size distribution was controlled with high pressure homogenization (1500 bar) and extrusion through polycarbonate membrane (3 and 0.8 μm). Fish gelatin was adsorbed to the interface to make secondary liposomes. Liposomal dispersions were osmotically dehydrated while monitoring the relative weight, size and rheological properties. The primary liposomes had an initial mean volume diameter (d_4,3) of 0.09, 0.40 and 2.7 μm and a ζ-potential of ?55 mV. Secondary liposomes were 0.11, 0.45 and 3.4 μm with a ζ-potential of 25 mV. The size of liposomes influenced the stability of liposomes, with the smallest liposomes being stable for 30 min, corresponding to 80 % of the initial weight, while the larger liposomes were already aggregated. Secondary liposomes were stable to 120 min for the smaller liposomes and to 150 min for the largest liposomes corresponding to 40 % of the initial weight. Stability increased during dehydration at 2 °C. Coating the liposomes increased the physical stability of the liposomal dispersions at all temperatures. The results show that cold water fish skin gelatin is a viable option to coat liposomes of a wide size range.     

Effect of temperature change on the tolerance of isolated contracting cardiac muscle to hypoxia.

The effects of graded temperature changes on tolerance to hypoxia are evaluated in isometrically contracting isolated muscle preparations from the rat. After equilibration at 28 °C, the temperature was changed to either 21 ° or 35 °C. Mechanical performance was measured after temperature change, during 60 min of hypoxia at a given temperature and a subsequent 30 min-period of reoxygenation at 28 °C. Performance as measured by (1) developed tension, (2) contracture tension, and (3) recovery on reoxygenation was diminished by raising temperature to 35 °C during hypoxia while no change in tolerance was demonstrated by lowering temperature from 28 ° to 21 °C.     

Toxic effects of DMSO on cultured beating heart cells at temperatures above zero

Toxic influences of DMSO were studied on spontaneously beating rat heart cells in tissue culture. Injurious effects of DMSO on the contractility and on the individual cell survival were investigated at three different temperatures, +4 ° C, +24 ° C, and + 37 ° C. As indicated by the 50% levels of irreversible damage it may be concluded that osmotic stress during the addition and the removal of DMSO should be avoided carefully at +4 ° C.With slow introduction and slow removal of DMSO optimal survival may be obtained up to 4.7 m DMSO at +4 ° C and at +24 ° C. Although at +37 ° C osmotic stress becomes less damaging, optimal survival can only be obtained at lower concentrations (up to 3.5 m depending on the incubation program). At +37 ° C the more specific toxic properties of DMSO are the limiting factor.At +24 ° C slow removal of DMSO instead of sudden removal at the end of an incubation period improves the survival considerably.At +24 ° C a good compromise between damage by osmotic shock and by other more specific toxic properties can be obtained.     

Effect of Indian mustard on feeding,larval survival and development of Plutella xylostella at constant temperatures

Feeding behavior of Plutella xylostella under optional to non-optional conditions was studied at 10°C, 15°C, 20°C, and 25°C on Indian mustard, Brassica juncea. The study reveals that the variety Pusa Bahar was significantly less preferred by the larvae as compared with Pusa Bold and Varuna under optional to non-optional conditions. Larvae of P. xylostella consumed more food at 25°C than 20°C, 15°C and 10°C. Larval survival was found to be highest on cabbage (control) as compared with Indian mustard and was found to vary with host plants and temperature. The larval survival decreased to 11.29% on Pusa Bahar at 10°C. Increasing the temperature from 10°C to 20°C, larval mortality resulted more on Varuna than Pusa Bahar and Pusa Bold. Developmental period was prolonged on Pusa Bold at 10°C while it was shortest on cabbage at 25°C. A total of 536.47 degree days were required to complete the development by immature stages on Varuna at 25°C and 421.64 degree days on cabbage.