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PubMed2004

Liposome-stabilized oil-in-water emulsions as adjuvants: increased emulsion stability promotes induction of cytotoxic T lymphocytes against an HIV envelope antigen.

Protective or therapeutic immunity against HIV infection is currently believed to require both antibody and CTL responses against the envelope protein. In the present study, the adjuvant activity of a unique oil-in-water emulsion, in which liposomes containing lipid A (LA) and encapsulated antigen served as the emulsifying agent, was examined in mice using oligomeric gp140 (ogp140) derived from the HIV-1 envelope as the antigen. Emulsions rendered either highly stable or unstable by altering the ratio of liposomes to oil were used to examine the effect of stability of the emulsion on adjuvant activity. Stable and unstable emulsions had similar potencies for inducing both IgG antibodies to ogp140 and antigen-specific T-lymphocyte proliferation. Stable emulsions, but not unstable emulsions, induced antigen-specific CTL responses, possibly because of the depot effect of the stable emulsions. Furthermore, stable emulsions induced lower IgG2a/IgG1 ratios than the unstable emulsions. We conclude that stable liposomal oil-in-water emulsions provide an effective means of obtaining both antibody and CTL responses against an HIV envelope antigen.

AIDS Vaccines↗
PubMed1985

Total intravenous anaesthesia in the horse with propofol.

The use of propofol, solubilised in a non-ionic emulsifying agent, for the induction and maintenance of anaesthesia in experimental ponies was assessed. Pilot studies revealed that premedication with xylazine (0.5 mg/kg bodyweight [bwt]) intravenously (iv) followed by propofol (2.0 mg/kg bwt) iv provided a satisfactory smooth induction. Two infusion rates (0.15 mg/kg bwt/min and 0.2 mg/kg bwt/min) were compared for maintenance of anaesthesia. An infusion rate of 0.2 mg/kg/min produced adequate anaesthesia in these ponies. Cardiovascular changes included a decrease in arterial pressure and cardiac output during maintenance. Respiratory depression was manifested by a decrease in rate and an increase in arterial carbon dioxide tension. Recovery after 1 h anaesthesia was rapid and smooth. In conclusion, induction and maintenance of anaesthesia with propofol in premedicated ponies proved a satisfactory technique.

Anesthesia, Intravenous↗
PubMed1988

The mannoprotein of Saccharomyces cerevisiae is an effective bioemulsifier.

The mannoprotein which is a major component of the cell wall of Saccharomyces cerevisiae is an effective bioemulsifier. Mannoprotein emulsifier was extracted in a high yield from whole cells of fresh bakers' yeast by two methods, by autoclaving in neutral citrate buffer and by digestion with Zymolase (Miles Laboratories; Toronto, Ontario, Canada), a beta-1,3-glucanase. Heat-extracted emulsifier was purified by ultrafiltration and contained approximately 44% carbohydrate (mannose) and 17% protein. Treatment of the emulsifier with protease eliminated emulsification. Kerosene-in-water emulsions were stabilized over a broad range of conditions, from pH 2 to 11, with up to 5% sodium chloride or up to 50% ethanol in the aqueous phase. In the presence of a low concentration of various solutes, emulsions were stable to three cycles of freezing and thawing. An emulsifying agent was extracted from each species or strain of yeast tested, including 13 species of genera other than Saccharomyces. Spent yeast from the manufacture of beer and wine was demonstrated to be a possible source for the large-scale production of this bioemulsifier.

Emulsions↗
PubMed2006

Stable concentrated emulsions of the 1-monoglyceride of capric acid (monocaprin) with microbicidal activities against the food-borne bacteria Campylobacter jejuni, Salmonella spp., and Escherichia coli.

Of 11 fatty acids and monoglycerides tested against Campylobacter jejuni, the 1-monoglyceride of capric acid (monocaprin) was the most active in killing the bacterium. Various monocaprin-in-water emulsions were prepared which were stable after storage at room temperature for many months and which retained their microbicidal activity. A procedure was developed to manufacture up to 500 ml of 200 mM preconcentrated emulsions of monocaprin in tap water. The concentrates were clear and remained stable for at least 12 months. They were active against C. jejuni upon 160- to 200-fold dilution in tap water and caused a >6- to 7-log(10) reduction in viable bacterial count in 1 min at room temperature. The addition of 0.8% Tween 40 to the concentrates as an emulsifying agent did not change the microbicidal activity. Emulsions of monocaprin killed a variety of Campylobacter isolates from humans and poultry and also killed strains of Campylobacter coli and Campylobacter lari, indicating a broad anticampylobacter activity. Emulsions of 1.25 mM monocaprin in citrate-lactate buffer at pH 4 to 5 caused a >6- to 7-log(10) reduction in viable bacterial counts of Salmonella spp. and Escherichia coli in 10 min. C. jejuni was also more susceptible to monocaprin emulsions at low pH. The addition of 5 and 10 mM monocaprin emulsions to Campylobacter-spiked chicken feed significantly reduced the bacterial contamination. These results are discussed in view of the possible utilization of monocaprin emulsions in controlling the spread of food-borne bacteria from poultry to humans.

Animal Feed↗
PubMed1983

Inhibition of bacterial adherence to hydrocarbons and epithelial cells by emulsan.

Acinetobacter calcoaceticus RAG-1 and BD413, as well as Streptococcus pyogenes M-5, adhered to octane. Adherence was inhibited by emulsan (100 micrograms/ml), the polymeric emulsifying agent produced by A. calcoaceticus RAG-1. Emulsan also inhibited adherence of S. pyogenes and RAG-1 to buccal epithelial cells. The mean values of bound S. pyogenes per epithelial cell were 57.2 and 20.7 for the control and emulsan-containing suspensions, respectively; mean values of bound RAG-1 per epithelial cell were 221 for the control and 40 for the suspension containing 100 micrograms of emulsan per ml. Desorption of previously bound RAG-1 from epithelial cells by emulsan was concentration dependent: a maximum of 80% desorption was obtained with 200 micrograms of emulsan per ml. The data showing that emulsan desorbed 70% of the indigenous bacterial flora from buccal epithelial cells suggest that hydrophobic interactions mediate not only the in vitro adherence of laboratory strains to epithelial cells, but actually govern the adherence of the majority of the bacteria that colonize this surface. The advantages of using emulsan as an antiadherence agent include its chemical purity, stability, and polymeric nature.

Acinetobacter↗
PubMed1994

Maternal transmission of immunity to Eimeria maxima: enzyme-linked immunosorbent assay analysis of protective antibodies induced by infection.

Vaccination of broiler chickens against Eimeria infection is problematic because of the need to ensure that birds are protected from the time of hatching. We have therefore investigated the feasibility of protecting hatchling broilers via maternal transfer of protective antibodies from hens to their offspring. Oral infection of broiler breeder hens with 20,000 sporulated Eimeria maxima oocysts caused production of antibodies which were passed into the egg yolk and subsequently to hatchlings. The level of specific antibodies in the yolks to unsporulated oocysts, sporulated oocysts, merozoites, and gametocytes was assessed by enzyme-linked immunosorbent assays. The levels in yolks of antibodies to all developmental stages peaked 3 to 4 weeks after infection of the hens. Groups of 10 hatchlings were challenged at 3 days of age by oral infection with 100 sporulated E. maxima oocysts. In the first experiment, the mean 4-day (days 6 to 9 post-infection) total number of oocysts excreted in the feces of chicks from eggs collected 3 weeks after infection of the hens was (0.6 +/- 0.4) x 10(6) (mean +/- standard error) compared with (9.9 +/- 1.4) x 10(6) for the progeny of uninfected hens, which represents a greater than 90% reduction. However, oocyst excretion by chicks from eggs collected 7 or 8 weeks after infection of the hens was only 47 or 68% lower than control values, reflecting declining levels of protective antibodies. In a second experiment, in which the hens were somewhat older and pretreated by intramuscular injection of saline in the emulsifying agent, Arlacel A, the period for which protective antibodies were transferred to hatchlings was prolonged. Thus, oocyst excretion by challenged hatchlings from eggs collected for an 8-week period after infection of the hens was more than 90% lower than oocyst excretion by control chicks, and even hatchlings of eggs collected 19 weeks after infection of the hens showed a 60% reduction in oocyst output. In both experiments, the levels of immunoglobulin G (IgG) antibodies to all developmental stages in yolks or hatchling sera were very strongly correlated with maternally derived immunity to E. maxima. In contrast, parasite-specific IgM or IgA was not detectable, either in egg yolk or egg white. These results demonstrate the ability of IgG antibodies to protect against E. maxima in poultry, thus raising the possibility of using protective maternally derived IgG antibodies to identify potentially protective parasite antigens and indicating the feasibility of using maternal immunization as a means for parasite control.

Animals↗
PubMed1981

Role of adherence in growth of Acinetobacter calcoaceticus RAG-1 on hexadecane.

The high affinity of Acinetobacter calcoaceticus RAG-1 for liquid hydrocarbons permitted the isolation of a spontaneous nonadherent mutant, MR-481. Strain MR-481 exhibited no significant affinity for three test hydrocarbons, yet resembled the wild type in many properties, including production of the extracellular emulsifying agent emulsan. To study the role of adherence in growth on hydrocarbons, RAG-1 and MR-481 were compared for growth on hexadecane under conditions of limited agitation and at low initial cell densities. Adherent RAG-1 cells were able to grow rapidly under these conditions, whereas nonadherent MR-481 cells failed to grow for at least 54 h. However, the addition of emulsan either initially or at various times after inoculation enabled the nonadherent MR-481 cells to grow on hexadecane. Growth was not the result of reversion of MR-481 from nonadherent to adherent cells. The data demonstrate that adherence is a crucial factor in the growth of A. calcoaceticus RAG-1 on hexadecane in the absence of extracellular emulsification of the substrate.

Acinetobacter↗
PubMed1984

Specific binding of a bacteriophage at a hydrocarbon-water interface.

Emulsan, the extracellular polyanionic emulsifying agent produced by Acinetobacter calcoaceticus RAG-1, has been implicated as a receptor for a specific virulent RAG-1 bacteriophage, ap3. Aqueous solutions of emulsan did not interfere with phage ap3 adsorption to RAG-1 cells. However, binding of phage ap3 occurred at the interfaces of hexadecane-in-water emulsions specifically stabilized by emulsan polymers. Binding of ap3 to emulsions was inhibited either in the presence of anti-emulsan antibodies or in the presence of a specific emulsan depolymerase. Moreover, when the phage was first bound to emulsan-stabilized emulsions and the emulsions subsequently treated with emulsan depolymerase, viable phage was released, indicating that phage ap3 DNA ejection was not triggered by binding. The results indicate that emulsan functions as the ap3 receptor and suggest that to function as a receptor, emulsan assumes a specific conformation conferred on it by its specific interaction with hydrophobic surfaces.

Acinetobacter↗
PubMed2002

Hydrophobicity development, alkane oxidation, and crude-oil emulsification in a Rhodococcus species.

The relationship between the phenomena alkane oxidation, extreme hydrophobicity of the cell surface, and crude-oil emulsification in Rhodococcus sp. strain 094 was investigated. Compounds that induce the emulsifying ability simultaneously induced the cytochrome P450-containing alkane oxidizing system and the transition from low to high cell-surface hydrophobicity. Exposed to inducers of crude-oil emulsification, the cells developed a strong hydrophobic character during exponential growth, which was rapidly lost when entering stationary phase. The loss in hydrophobicity coincided in time with the crude-oil emulsification, indicating that the components responsible for the formation of cell-surface hydrophobicity act as excellent emulsion stabilisers only after release from the cells. Rhodococcus sp. strain 094 possessed three distinct levels of cell-surface hydrophobicity. One level of low hydrophobicity was characteristic of cells in late stationary phase and was independent of growth substrate. A second and more hydrophobic level was observed for cells in exponential phase grown on water-soluble substrates, while a third level, characterised by extreme cell hydrophobicity, was observed for cells in exponential phase cultivated on hydrophobic substrates such as hexadecane. The production of the oil-emulsifying agents seems to require external sources of nitrogen and phosphate.

Alkanes↗
PubMed1982

Aerial spraying of fenitrothion in forest programs: some problems and some solutions.

Annually, large tracts of forest in eastern Canada are sprayed aerially with insecticides (fenitrothion, aminocarb) in attempts to control an epidemic infestation by an indigenous forest pest, the spruce budworm (Choristoneura fumiferana, Clemens). The massive size of the spraying programs, the anecdotal reports of human exposure, and the potential for hazard to human health have led one province. New Brunswick, to initiate and fund specific environmental and laboratory studies which will provide adequate data upon which the government can base realistic legislation to protect both the forests and the population. These studies have included some unique field analyses of aerial spray drift conducted by a research group from the National Research Council; comparative subchronic studies in rats of fenitrothion and a new formulation; a nose-only inhalation study of this formulation in rats; field testing of the formulation for drift characteristics. Ongoing research involves the subchronic testing of the emulsifying agents being used routinely in the new formulation and studies of a low-drift additive which will stabilize the particle size of the spray. On the basis of the results to date, the government has been able to modify spraying techniques and to establish realistic buffer zones around human habitation.

Aerosols↗
PubMed1999

Effects of a perfluorochemical emulsion on the fate of circulating Pseudomonas aeruginosa.

Because mononuclear phagocytes take up perfluorochemical emulsions (PFCE), we examined how prior treatment with PFCE affects the fate of circulating bacteria. Rats were preinjected with three daily intravenous injections of PFCE (2.0 ml/100 g) containing 12.5% (vol/vol) of a 4:1 mixture of F-dimethyl adamantane and F-trimethylbicyclo-nonane, 2.5% (wt/vol) Pluronic F-68 as the emulsifying agent, and 3% (wt/vol) hydroxyethyl starch as the oncotic agent. Pseudomonas aeruginosa or Staphylococcus aureus were injected 4 h after the third PFCE injection. PFCE pretreatment decreased the rate and extent of vascular clearance of P. aeruginosa, with decreased uptake by the liver. Importantly, there were significant decreases in killing of P. aeruginosa in the liver, lungs, spleen, and kidneys of PFCE animals. PFCE did not alter the clearance of S. aureus from the circulation. However, hepatic uptake was reduced, with concomitant increases in lung and kidney uptake. Ultrastructure of Kupffer cells revealed PFCE inclusions and extensive vacuolization. These experiments demonstrate that the clearance kinetics and organ distribution of circulating P. aeruginosa and their subsequent killing are altered by PFCE. Diminished hepatic phagocyte function leads to a decrease in vascular clearance of circulating bacteria, increased uptake in other reticuloendothelial organs, and decreased bactericidal activity versus P. aeruginosa.

Animals↗
PubMed1992

Increased plasma O2 solubility improves O2 uptake of in situ dog muscle working maximally.

A perfluorocarbon emulsion [formulation containing 90% wt/vol perflubron (perfluorooctylbromide); Alliance Pharmaceutical] was used to increase O2 solubility in the plasma compartment during hyperoxic low hemoglobin concentration ([Hb]) perfusion of a maximally working dog muscle in situ. Our hypothesis was that the increased plasma O2 solubility would increase the muscle O2 diffusing capacity (DO2) by augmenting the capillary surface area in contact with high [O2]. Oxygen uptake (VO2) was measured in isolated in situ canine gastrocnemius (n = 4) while working for 6 min at a maximal stimulation rate of 1 Hz (isometric tetanic contractions) on three to four separate occasions for each muscle. On each occasion, the last 4 min of the 6-min work period was split into 2 min of a control treatment (only emulsifying agent mixed into blood) and 2 min of perflubron treatment (6 g/kg body wt), reversing the order for each subsequent work bout. Before contractions, the [Hb] of the dog was decreased to 8-9 g/100 ml and arterial PO2 was increased to 500-600 Torr by having the dog breathe 100% O2 to maximize the effect of the perflubron. Muscle blood flow was held constant between the two experimental conditions. Plasma O2 solubility was almost doubled to 0.005 ml O2 x 100 ml blood-1 x Torr-1 by the addition of the perflubron. Muscle O2 delivery and maximal VO2 were significantly improved (at the same blood flow and [Hb]) by 11 and 12.6%, respectively (P < 0.05), during the perflubron treatment compared with the control. O2 extraction by the muscle remained the same between the two treatments, as did the estimate of DO2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

Differential contributions of major lipid components of atheroma to outcome of cerebral atheroembolism. A study in an animal model.

Cerebral atheroembolism, in which mainly lipid emboli are released from rupturing atheromatous plaques, may occur without apparent effect, or result in cerebral ischemia and infarction. The reasons behind these unpredictable consequences were sought in the interaction, in vitro and in an animal model, between the main lipid components of advanced plaques. Pure preparations of representative lipids were each harmless when embolized into the cerebral circulation. In contrast, combinations in proportions similar to those in advanced human plaques caused infarction, whether these were synthetic mixtures or extracts from plaques of the entire lipid fraction. The most important physical interaction between the lipids was aggregation of crystals by oils. Between cholesterol and the mainly liquid esters, this created in vitro a range of glutinous aggregates. Triglyceride lowered the melting point of esters, increasing their oiliness, and reduced the cohesiveness of aggregates in the face of operative mechanical forces through a fall in viscosity. Phospholipid, acting principally as an emulsifying agent, promoted dispersion of the oil, secondarily freeing the crystals from its aggregating effect. In the plaque, the balance of these factors will determine the size and number of particles likely to embolize, and, therefore, the clinical outcome should the plaque rupture.

Animals↗
PubMedDate not supplied

Pharmacologic levels of heparin do not destabilize neonatal parenteral nutrition.

BACKGROUND: Calcium and heparin are known to destabilize the lipid emulsion of total parenteral nutrition (TPN). However, these observations were made over long periods of time, using 5 to 10 times the amount of heparin used in a neonatal intensive care unit. We investigated the effects of lower heparin concentrations with lipid-to-nutrient ratios normally administered to premature infants. METHODS: Lipid emulsion stability was assessed over 30 minutes by measuring viscosity at 10 mmol/L calcium after the addition of 0, 0.5, 1, and 5 IU heparin/mL. This was done at a range of lipid-to-nutrient ratios in clinical use. The effect of varying calcium concentration and different multivitamin preparations on emulsion stability also was observed. RESULTS: Heparin caused an immediate increase in viscosity of pure Intralipid 20% (Intralipid; Kabi Pharmacia AB, Stockholm, Sweden), which eventually separated into two phases. Although changes in viscosity were observed at 1:1 lipid-to-nutrient ratios, no effect was seen at a 1:9 ratio. With the 1:1 ratio, the multivitamin preparations, MVI Pediatric (Rhone-Poulec Rorer, Montreal, Canada) and Vitlipid (Kabi Pharmacia AB), reduced the increase in viscosity. CONCLUSIONS: Heparin and calcium destabilize Intralipid. This is unlikely to cause problems for most infants receiving TPN, provided low heparin concentrations are used. In all cases, especially where the lipid ratio is high, the risk of the lipid phase separating out can be further minimized by (1) the addition of multivitamin preparations containing detergent or an emulsifying agent and (2) by having the shortest possible delivery tube between the point of mixing the lipid and amino acid solutions of TPN and entry into the infant.

Calcium↗
PubMedDate not supplied

Cytotoxicity of amino alcohols to rat hepatoma-derived Fa32 cells.

Amino alcohols are used as emulsifying agents in dry-cleaning soaps, wax removers, cosmetics, paints and insecticides. The cytotoxicities of 12 amino alcohols, which differed in chain length, position of the amino and alcohol groups, and the presence of an additional phenyl group, were determined by the neutral red uptake inhibition assay with normally cultured, glutathione-depleted or antioxidant-enriched Fa32 rat hepatoma-derived cells. Glutathione depletion and antioxidant enrichment were achieved by including 50(M L-buthionine-S,R-sulphoximine (BSO) or 100(M (-tocopherol acetate (vitamin E) in the culture medium for 24 hours before and during the assay. The cytotoxicity of the amino alcohols observed after treatment for 24 hours was expressed as the concentration of compound needed to induce a 50% reduction in neutral red uptake (NI50). The observed NI50 values ranged from 3mM to 30mM. The individual stereoisomers and a racemic mixture of 1-amino-2-propanol exhibited similar cytotoxicities (with normally cultured Fa32 cells, and vitamin E- and BSO-treated cultures). Similar NI50 values for D-(+)-2-amino-1-propanol, 3-amino-1-propanol and the L-, D- or DL- forms of 1-amino-2-propanol, indicated that the position of the amino group had little influence on the cytotoxicities of the amino alcohols. In contrast, the position of the hydroxyl group appeared to play an important role for the toxicity of the compound, as indicated by the significantly different NI50 values for 4-amino-1-butanol and 4-amino-2-butanol. An additional phenyl group greatly increased the cytotoxicity of 2-amino-1,3-propanediol. For most of the compounds, cytotoxicity increased when GSH was depleted, and decreased when the cells were enriched with vitamin E. This indicated that most of the tested chemicals interact with GSH, either directly or indirectly, by processes which generate oxygen free-radicals. Decreased toxicity was found for most of the chemicals administered to vitamin E-enriched cells, indicating that reactive oxygen species could be involved in the toxicity of the amino alcohols.

Amino Alcohols↗
PubMed1985

Anaphylactoid reactions associated with parenteral cyclosporine use: possible role of Cremophor EL.

Acute anaphylactoid reactions occurred immediately after initiation of intravenous infusions of cyclosporine in three patients post-organ transplantation. Shortness of breath, flushing, tachypnea, chest pain, pruritus, or urticaria were noted; rapid recovery followed cessation of drug infusion. Subsequently, oral cyclosporine has been used in each patient without recurrence of the observed reaction. The presence of Cremophor EL as an emulsifying agent in the parenteral dosage formulation of cyclosporine is a likely etiology for this acute adverse reaction. Slowed rates of drug infusion and antihistamine premedication may permit continued intravenous cyclosporine use in affected patients.

Adult↗
PubMed1991

Octenylsuccinic aciduria in children fed protein-hydrolysate formulas containing modified cornstarch.

The excretion of 2-(2'-octenyl)succinic acid (OSA) and several metabolites of OSA was studied by gas chromatography/mass spectrometry in 17 infants and children fed one of three proprietary elemental or protein-hydrolysate formulas that use OSA-modified cornstarch as an emulsifying agent. Variable but often large amounts (up to 2500 mg/g creatinine) of the fatty acid-like OSA and its metabolites were found in the urine of these children, and levels of OSA in their blood ranged from 9.5 to 57.9 mumol/L. Apparently secondary abnormalities, such as increased urinary levels of glutaric acid and 2-ketoglutaric acid, were also found in more than half of the urine specimens. The molecular weight and mass fragmentation patterns of the nine compounds associated with the excretion of OSA are consistent with the proposal that OSA is metabolized in human infants and children by a combination of omega-, omega-1-, and beta-oxidation steps, similar to the metabolism of another branched-chain fatty acid, valproic acid. The urinary organic acid pattern of children fed elemental formulas containing OSA-modified starch often was dominated by OSA and its metabolites, and in several children the OSA-related changes were mistaken for a primary metabolic disease. Physicians and laboratories evaluating children for suspected metabolic diseases should be aware of the possibility of abnormal organic acid studies associated with OSA-containing formulas.

Child↗
PubMed1990

Intravascular behavior of a perfluorochemical emulsion.

The purpose of this study was to clarify the reason why two similar perfluorochemical (PFC) emulsions, namely a mixed PFC (perfluorodecalin: FDC and perfluorotripropylamine: FTPA) and an FDC emulsion, resulted in a very different survival time for the exchange-transfused rats. Supposing that some difference in the intravascular behavior of both emulsions would account for such a difference in efficacy, experiments on behavior of PFC emulsions were carried out focusing on the particle size. It was reconfirmed that larger PFC particles were eliminated from the blood stream much more rapidly than smaller particles with three FMIQ (perfluoro-N-methyldecahydroisoquinoline) emulsions. After the FDC + FTPA emulsion or the FDC emulsion were injected into rabbits, PFC particles in the blood tended to decrease in size. When each of the collected blood samples was incubated at 37 degrees C for 24 h, the FDC emulsion enlarged in size markedly, but the FDC + FTPA emulsion showed no change. The retention of PFC particles appeared to depend on the emulsion stability rather than simply on the emulsifying agent alone. These data showed that some differences were observed in intravascular persistence of the FDC + FTPA emulsion and the FDC emulsion, and suggested that the efficacy of PFC emulsions would reflect their behavior in the circulation.

Animals↗