RESUMEN
BACKGROUND: Oral health promotion initiatives must be evidence-based and consistent with broader health messaging. The Oral Health Messages for the Australian Public were first produced in 2009 and sought to enable a focused, and strategic approach to oral health promotion in Australia. As the evidence base and needs of the Australian population have since changed, this consensus statement was updated in 2022-2023. METHODS: The process of updating the messages consisted of 3 phases (preparatory phase, the Delphi technique, final revision phase). The preparatory phase included public and expert consultation, an umbrella review of published scientific literature and review of available recommendations, policies and guidelines. The Delphi technique used in this study was guided by Guidance on Conducting and REporting DElphi Studies (CREDES) and included 2 voting rounds. There were 70 experts in round 1 and 60 experts in round 2. Delphi participants comprised of experts from a variety of fields to ensure diversity and inclusion, balance expertise and maximize stakeholder representation. Consensus was defined as 75% agreement. RESULTS: A total of 11 messages were included in the 2022 update of Oral Health Messages for Australia. CONCLUSION: The updated oral health messages will support oral health promotion policy and activity at both individual and population level to improve the oral health of Australians.
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Promoción de la Salud , Salud Bucal , Humanos , Australia , Promoción de la Salud/métodos , Consenso , Técnica DelphiRESUMEN
BACKGROUND: High temperature is a major abiotic stress that limits wheat (Triticum aestivum L.) productivity. Variation in levels of a wide range of lipids, including stress-related molecular species, oxidative damage, cellular organization and ultrastructural changes were analyzed to provide an integrated view of the factors that underlie decreased photosynthetic rate under high temperature stress. Wheat plants of cultivar Chinese Spring were grown at optimum temperatures (25/15 °C, maximum/minimum) until the onset of the booting stage. Thereafter, plants were exposed to high temperature (35/25 °C) for 16 d. RESULTS: Compared with optimum temperature, a lower photosynthetic rate was observed at high temperature which is an interplay between thylakoid membrane damage, thylakoid membrane lipid composition, oxidative damage of cell organelle, and stomatal and non-stomatal limitations. Triacylglycerol levels were higher under high temperature stress. Polar lipid fatty acyl unsaturation was lower at high temperature, while triacylglycerol unsaturation was the same at high temperature and optimum temperature. The changes in lipid species indicates increases in activities of desaturating, oxidizing, glycosylating and acylating enzymes under high temperature stress. Cumulative effect of high temperature stress led to generation of reactive oxygen species, cell organelle and membrane damage, and reduced antioxidant enzyme activity, and imbalance between reactive oxygen species and antioxidant defense system. CONCLUSIONS: Taken together with recent findings demonstrating that reactive oxygen species are formed from and are removed by thylakoid lipids, the data suggest that reactive oxygen species production, reactive oxygen species removal, and changes in lipid metabolism contribute to decreased photosynthetic rate under high temperature stress.
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Fotosíntesis/fisiología , Triticum/metabolismo , Calor , Oxidación-Reducción , Fotosíntesis/genética , Especies Reactivas de Oxígeno/metabolismo , Temperatura , Triglicéridos/metabolismo , Triticum/genéticaRESUMEN
High temperature (HT) decreases seed set percentage in sorghum (Sorghum bicolor [L.] Moench). The relative sensitivity of pollen and particularly pistil and the mechanistic response that induces tolerance or susceptibility to HT are not well known and hence are the major objectives of this research. The male sterile (ATx399) and fertile (RTx430) lines were exposed to 30/20 °C (optimum temperature), 36/26 °C (HT1 ), and 39/29 °C (HT2 ) from the start of booting to seed set in a controlled environment. Similarly, in the field, HT stress was imposed using heat tents. HT stress decreased pollen germination. Relatively high levels of reactive oxygen species and decreased antioxidant enzyme activity and phospholipid unsaturation were observed in pollen compared to pistil under HT. The severe cell organelle damage was observed in pollen and pistil at 36/26 and 39/29 °C, respectively. The seed set percentage was higher in HT-stressed pistil pollinated with optimum-temperature pollen. Direct and reciprocal crosses indicate that pollen was more sensitive with larger decreases in seed set percentage than pistil under HT stress. The negative impact was greater in pollen than pistil at lower temperatures. Overall, pollen was more sensitive than pistil to HT stress because it is more susceptible to oxidative damage than pistil.
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Antioxidantes/metabolismo , Flores/fisiología , Especies Reactivas de Oxígeno/metabolismo , Sorghum/fisiología , Flores/ultraestructura , Calor , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Fosfolípidos/metabolismo , Polen/fisiología , Polen/ultraestructura , Polinización , Sorghum/ultraestructura , Estrés FisiológicoRESUMEN
Although serum albumin is the major protein component of the aqueous humor and vitreous humor, its possible physiological role(s) in lens metabolism has not yet been determined. BODIPY fatty acid, a fluorescent analogue of a C(12) long chain fatty acid, was bound to serum albumin, then incubated with lenses in culture. After various times of incubation, the lenses were homogenized, lipids extracted, and the extracted lipids resolved by thin layer chromatography. Fluorescence analyses demonstrated that the BODIPY fatty acid--albumin complex was translocated into the lens, where the BODIPY fatty acid was incorporated in a time dependent manner into numerous lipids, including phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin. Together, the results strongly suggest that serum albumin in the aqueous humor and/or vitreous humor facilitates the translocation of long chain fatty acids into the lens, where they are used for the biosynthesis of lens lipids.
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Ácidos Grasos/metabolismo , Cristalino/metabolismo , Lípidos/biosíntesis , Albúmina Sérica/metabolismo , Animales , Compuestos de Boro/metabolismo , Células Cultivadas , Colorantes Fluorescentes/metabolismo , Masculino , Microscopía Confocal/métodos , Técnicas de Cultivo de Órganos/métodos , Fosfatidilcolinas/biosíntesis , Fosfatidiletanolaminas/biosíntesis , Ratas , Esfingomielinas/biosíntesisRESUMEN
The common plant phospholipase D (PLD), PLDalpha, has been proposed to be involved in wound-induced production of jasmonic acid. To better understand the role(s) of PLDalpha in the wound response, detailed lipid analysis was carried out to determine the in vivo substrates and the contribution of PLDalpha in wound-induced lipid metabolism in Arabidopsis thaliana. Mechanical wounding of Arabidopsis leaves resulted in significantly less hydrolysis of phosphatidylcholine (PC) in PLDalpha-deficient than in wild-type plants. Hydrolysis of phosphatidylethanolamine, phosphatidylglycerol (PG), and phosphatidylinositol within 30 min of wounding was not significantly different in PLDalpha-deficient and wild-type leaves. Phosphatidic acid (PA) levels increased rapidly in wild-type and, to a lesser extent, in PLDalpha-deficient plants. The acyl composition of the PA generated by wounding suggests that the major in vivo substrate of PLD in wild-type leaves was PC, and that PG hydrolysis accounted for 10-15% of the wound-induced PA in wild-type leaves. Comparison of the acyl compositions of the wound-induced PA of wild-type and PLDalpha-deficient leaves indicated that PLDalpha hydrolyzed PG more readily than other PLD isoforms did. Wounding produced substantial increases in free linoleic and linolenic acids in wild-type plants, whereas PLDalpha-deficient plants showed only a slight increase in linoleic acid and no significant increase in linolenic acid. These results demonstrate that PLDalpha and at least one other PLD isoform, as well as other hydrolytic enzymes, are active in mechanically wounded Arabidopsis leaves, and PLDalpha is involved in wound-induced metabolism of polyunsaturated fatty acids.
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Arabidopsis/metabolismo , Fosfolipasa D/metabolismo , Fosfolípidos/metabolismo , Arabidopsis/genética , Diglicéridos/metabolismo , Ácidos Fosfatidicos , Fosfatidilcolinas/metabolismo , Fosfolipasa D/deficiencia , Especificidad por Sustrato , Ácido alfa-Linolénico/metabolismoRESUMEN
Multiple forms of phospholipase D (PLD) were activated in response to wounding, and the expressions of PLDalpha, PLDbeta, and PLDgamma differed in wounded Arabidopsis leaves. Antisense abrogation of the common plant PLD, PLDalpha, decreased the wound induction of phosphatidic acid, jasmonic acid (JA), and a JA-regulated gene for vegetative storage protein. Examination of the genes involved in the initial steps of oxylipin synthesis revealed that abrogation of the PLDalpha attenuated the wound-induced expression of lipoxygenase 2 (LOX2) but had no effect on allene oxide synthase (AOS) or hydroperoxide lyase in wounded leaves. The systemic induction of LOX2, AOS, and vegetative storage protein was lower in the PLDalpha-suppressed plants than in wild-type plants, with AOS exhibiting a distinct pattern. These results indicate that activation of PLD mediates wound induction of JA and that LOX2 is probably a downstream target through which PLD promotes the production of JA.
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Arabidopsis/metabolismo , Ciclopentanos/metabolismo , Fosfolipasa D/metabolismo , Enfermedades de las Plantas , Arabidopsis/enzimología , OxilipinasRESUMEN
We report the complete sequence of an extreme halophile, Halobacterium sp. NRC-1, harboring a dynamic 2,571,010-bp genome containing 91 insertion sequences representing 12 families and organized into a large chromosome and 2 related minichromosomes. The Halobacterium NRC-1 genome codes for 2,630 predicted proteins, 36% of which are unrelated to any previously reported. Analysis of the genome sequence shows the presence of pathways for uptake and utilization of amino acids, active sodium-proton antiporter and potassium uptake systems, sophisticated photosensory and signal transduction pathways, and DNA replication, transcription, and translation systems resembling more complex eukaryotic organisms. Whole proteome comparisons show the definite archaeal nature of this halophile with additional similarities to the Gram-positive Bacillus subtilis and other bacteria. The ease of culturing Halobacterium and the availability of methods for its genetic manipulation in the laboratory, including construction of gene knockouts and replacements, indicate this halophile can serve as an excellent model system among the archaea.
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Genoma Bacteriano , Halobacterium/genética , Evolución Biológica , Membrana Celular/metabolismo , Reparación del ADN , Replicación del ADN , Metabolismo Energético , Halobacterium/metabolismo , Membrana Dobles de Lípidos , Datos de Secuencia Molecular , Biosíntesis de Proteínas , Recombinación Genética , Transducción de Señal , Transcripción GenéticaRESUMEN
The twisting ability of a series of 1,1'-binaphthalene compounds used as dopants in nematic solvents has been related to the dihedral angle theta between the two naphthalene moieties. While in the case of the more flexible compounds the sign and value of the helical twisting power is affected by several structural features that prevent a simple assignment of the conformation, in the presence of a covalent bridge that restricts the rotation around the C(1)-C(1') bond a reliable estimate of the conformational helicity could be obtained. This technique is complementary to CD spectroscopy that, for the investigated molecules, presents the same exciton patterns irrespective of the actual theta value.
RESUMEN
The dansyl fluors of 1-oleoyl-2-[4-(dansyl)amino]butyl]-sn-glycero-3-phosphocholine (18:1,4-dansyl PC) and 1-palmitoyl-2-[1-[(dansyl)amino)]undecanoyl]-sn-glycero-3-phosphoc holine (16:0,11-dansyl PC) were shown to reside at similar depths in phosphatidylcholine vesicles at pH 7. Analysis of fluorescence emission maxima showed that the dansyl groups of both 18:1,4-dansyl PC and 16:0,11-dansyl PC in phosphatidylcholine vesicles experienced environments more polar than methanol, suggesting that the dansyl group attached to the terminus of the undecanoyl chain must fold back toward the bilayer surface. Fluorescence polarization measurements in solid/fluid lipid mixtures show that both probes partition strongly into fluid phase lipid. The very low polarization values of 16:0,11-dansyl PC in lipid vesicles are consistent with the notion that the dansyl fluor of 16:0,11-dansyl PC existed in an environment allowing a high degree of motional freedom due to folding back of the dansyl group attached to the undecanoyl chain.
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Compuestos de Dansilo/química , Membrana Dobles de Lípidos/química , Fosfatidilcolinas/química , Compuestos de Dansilo/síntesis química , Polarización de Fluorescencia , Colorantes Fluorescentes , Conformación Molecular , Fosfatidilcolinas/síntesis química , Marcadores de SpinRESUMEN
To investigate the role of iron, ascorbate, and fructose on copper depletion and the effect of copper depletion on neonatal lung collagen, elastin, and surfactant, female rabbits were fed a control diet [10 parts per million (ppm) copper], a basal marginal copper diet (1.5 ppm), or a basal diet containing a high concentration of iron (1,750 ppm), ascorbic acid (1%, wt/wt), or fructose (20% of carbohydrates, wt/wt) or a combination of iron, ascorbic acid, and fructose throughout gestation. Whereas 10% of neonates in the control group died in the first 24 h, 27-67% of the offspring of rabbits fed the marginal copper diet died. Birth weight was also lower for the pups of the females fed the marginal copper diets. Lungs of neonates born to females fed iron or ascorbate and marginal copper diets had low levels of copper, high proportions of acid-extractable, high-molecular-weight collagen, and low lysyl-oxidase activities, consistent with incomplete maturation of collagen. The bronchoalveolar lavage fluids of newborns whose mothers were fed marginal copper diets alone or in combination with iron and/or ascorbate had lower levels of total surfactant phospholipids than the fluids from lungs of control newborns. The lower surfactant phospholipid content of these groups could be attributed mainly to lower phosphatidylcholine and, in particular, dipalmitoylphosphatidylcholine levels. These results suggest that high maternal intakes of iron, ascorbate, or their combination in pregnancy deplete biologically available copper, which in turn induces neonatal lung abnormalities.
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Animales Recién Nacidos/metabolismo , Colágeno/biosíntesis , Cobre/deficiencia , Elastina/biosíntesis , Pulmón/metabolismo , Preñez/metabolismo , Surfactantes Pulmonares/biosíntesis , Animales , Ácido Ascórbico/farmacología , Peso Corporal , Colágeno/análisis , Cobre/análisis , Elastina/análisis , Femenino , Hierro/análisis , Hierro/farmacología , Embarazo , Proteína-Lisina 6-Oxidasa/metabolismo , Surfactantes Pulmonares/análisis , ConejosRESUMEN
Lipid domains that occur within biological of model membranes encompass a variety of structures with very different lifetimes. The separation of membrane lipids into compositional domains can be due to lateral phase separation, immiscibility within a single phase, or interaction of lipids with integral or peripheral proteins. Lipid domains can affect the extent and rate of reactions in the membrane and provide sites for the activity of specialized proteins. Domains are likely to be involved in the process of lipid sorting to various cellular membranes, as well as in other processes which involve membrane budding or invagination.
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Lípidos de la Membrana/química , Lípidos de la Membrana/fisiología , Membranas Artificiales , Animales , Proteínas del Citoesqueleto/metabolismo , Fluidez de la Membrana , Modelos Biológicos , Conformación Molecular , Esteroles/química , Esteroles/metabolismoRESUMEN
Dimethylsuberimidate cross-linking has been used to determine the arrangement of phosphatidylethanolamine (PE) molecular species in Escherichia coli membranes. No large deviations from random mixing were found in wild-type strain AB1623, either in whole cells or in extracted lipids which were reconstituted into multilamellar vesicles. These results suggest, first, that there is little difference in the PE molecular species composition of the three lipid monolayers (the inner and outer monolayers of the inner membrane and the inner monolayer of the outer membrane) which contain significant amounts of PE. Secondly, the results suggest that the molecular species within each monolayer and in the extracted lipids are arranged close to randomly with no tendency for like molecular species to cluster. E. coli strain L8-2, which has a defect in beta-oxidation and a temperature-sensitive mutation in total fatty acid synthesis, was grown on cis-vaccenate (cis-11,12- octadecenate) to enrich the cells in divaccenoyl PE. Again, in whole cells or in lipids extracted from whole cells and reconstituted into multilamellar vesicles, the species were close to randomly arranged. However, a consistent, slight tendency of divaccenoyl species to pair with like species as compared to pairing with the second most common species, vaccenoyl, palmitoleoyl PE, was noted in both extracted lipids and in whole cells.
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Dimetil Suberimidato/química , Escherichia coli/química , Lípidos/química , Fosfatidiletanolaminas/química , Membrana Celular/química , Escherichia coli/genética , Concentración de Iones de HidrógenoRESUMEN
Membrane lipid compositions of Cryptosporidium parvum and Madin-Darby bovine kidney cells, an epithelial-like cell line commonly used to study coccidia in vitro, were analyzed using both thin-layer chromatography and gas-liquid chromatography. Phosphatidylcholine was the predominant lipid in both C. parvum and Madin-Darby bovine kidney cells, comprising 65% and 41% of the total phospholipids, respectively. Phospholipids of C. parvum contained twice the level of 16:0 and twenty-fold more 18:2 than the Madin-Darby bovine kidney cell line. We suggest that the parasite may be capable of sequestering specific complex membrane lipids at concentrations greater than those in the host cells. This study constitutes the first report of the lipid composition of C. parvum.
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Cryptosporidium parvum/química , Riñón/química , Lípidos/análisis , Animales , Bovinos , Línea Celular , Ácidos Grasos/análisis , Cabras , Riñón/citología , Lípidos/clasificación , Lípidos de la Membrana/análisis , Fosfolípidos/análisisRESUMEN
A specific plasma membrane glycosphingolipid, known as ganglioside GM3, can regulate the intrinsic tyrosyl kinase activity of the epidermal growth factor (EGF) receptor; this modulation is not associated with alterations in hormone binding to the receptor. GM3 inhibits EGF receptor tyrosyl kinase activity in detergent micelles, in plasma membrane vesicles, and in whole cells. In addition, immunoaffinity-purified EGF receptor preparations contain ganglioside GM3 (Hanai et al. (1988) J. Biol. Chem. 263, 10915-10921), implying that the glycosphingolipid is intimately associated with the receptor kinase in cell membranes. Both the nature of this association and the molecular mechanism of kinase inhibition remain to be elucidated. In this report, we describe the synthesis of a fluorescent analog of ganglioside GM3, in which the native fatty acid was replaced with trans-parinaric acid. This glycosphingolipid inhibited the receptor kinase activity in a manner similar to that of the native ganglioside. A modified fluorescent glycosphingolipid, N-trans-parinaroyl de-N-acetyl ganglioside GM3, was also prepared. This analog, like the nonfluorescent de-N-acetyl ganglioside GM3, had no effect on receptor kinase activity. Results from tryptophan fluorescence quenching and steady-state anisotropy measurements in membranes containing these fluorescent probes and the human EGF receptor were consistent with the notion that GM3, but not de-N-acetyl GM3, interacts specifically with the receptor in intact membranes.
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Receptores ErbB/metabolismo , Ácidos Grasos Insaturados/farmacología , Gangliósido G(M3)/análogos & derivados , Gangliósido G(M3)/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Células 3T3 , Animales , Secuencia de Carbohidratos , Línea Celular , Membrana Celular/metabolismo , Receptores ErbB/efectos de los fármacos , Ácidos Grasos/análisis , Ácidos Grasos Insaturados/metabolismo , Polarización de Fluorescencia , Gangliósido G(M3)/síntesis química , Gangliósido G(M3)/farmacología , Humanos , Indicadores y Reactivos , Cinética , Liposomas , Ratones , Micelas , Datos de Secuencia Molecular , Espectrometría de Fluorescencia , TermodinámicaRESUMEN
1-Octadecanoyl, 2-decanoylphosphatidylethanolamine (C(18:0)C(10:0)PE) has been reported to exhibit mixed interdigitated gel-phase packing of the phospholipid acyl chains (Mason, J.T. and Stephenson, F.A. (1990) Biochemistry 29, 590-598). In contrast, ditetradecanoylphosphatidylethanolamine (C(14:0)C(14:0)PE) packs without significant interdigitation of the phospholipid acyl chains across the bilayer center. In this report, the gel-fluid transition temperatures of C(18:0)C(10:0)PE and C(14:0)C(14:0)PE in multilamellar dispersions were determined by fluorescence anisotropy of cis-parinaric acid and trans-parinaric acid with a descending temperature scan rate of 0.67 degrees C/min. The transition mid-points detected for C(18:0)C(10:0)PE with cis-parinaric acid were 19 degrees C in water, 18 degrees C at pH 8.1, and 14 degrees C at pH 10. The phase diagram for C(14:0)C(14:0)PE and C(18:0)C(10:0)PE at pH 10 suggests complete mixing in the fluid phase and considerable immiscibility in the gel phase. Cross-linking of equimolar mixtures of C(14:0)C(14:0)PE and C(18:0)C(10:0)PE with dimethylsuberimidate at pH 10 revealed a random arrangement of the two species in the fluid phase, confirming the notion that C(18:0)C(10:0)PE and C(14:0)C(14:0)PE are miscible in the fluid phase, as determined from the phase diagram. In contrast, cross-linking of the equimolar mixture of C(18:0)C(10:0)PE and C(14:0)C(14:0)PE in the gel phase at 0 degrees C revealed a non-random arrangement, demonstrating and confirming immiscibility in the gel phase.
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Geles , Fosfatidiletanolaminas/química , Fenómenos Químicos , Química Física , Ácidos Grasos Insaturados/química , Polarización de Fluorescencia , Concentración de Iones de Hidrógeno , Estructura Molecular , EstereoisomerismoRESUMEN
Previously it was reported (Bremer, E.G., Schlessinger, J., and Hakomori, S.-I. (1986) J. Biol. Chem. 261, 2434-2440) that ganglioside GM3 inhibited epidermal growth factor (EGF)-stimulated phosphorylation of the EGF receptor in Triton X-100-treated preparations of human epidermoid carcinoma (A431) cell membranes. In addition, these authors reported that GM3 inhibited the growth of A431 cells. In contrast, a modified ganglioside, de-N-acetyl GM3, enhanced the EGF-dependent tyrosine kinase activity of the EGF receptor. In this work and in subsequent studies (Hanai, N., Dohi, T., Nores, G. A., and Hakomori, S.-I. (1988) J. Biol. Chem. 263, 6296-6301), the tyrosine kinase activity of the receptor from A431 cell membranes was assayed in the presence of Triton X-100. In this report, we confirm that GM3 inhibited and de-N-acetyl GM3 stimulated EGF receptor autophosphorylation in the presence of Triton X-100. However, in the absence of detergents, ganglioside GM3 inhibited EGF-stimulated receptor autophosphorylation, whereas de-N-acetyl GM3 had no effect on EGF-stimulated receptor autophosphorylation. The effects of these gangliosides on receptor autophosphorylation were measured in both A431 cell plasma membranes and in 3T3 cell membranes permeabilized to [32P]ATP by a freeze-thaw procedure, in intact A431 cells permeabilized with alamethicin, and in intact A431 cells grown in the presence of [32P]orthophosphate. Thus, the inhibitory effect of GM3 on receptor autophosphorylation was demonstrated in the presence and in the absence of detergent; the stimulatory effect of de-N-acetyl GM3 was observed only in the presence of detergent. We also demonstrate that ganglioside GM3 inhibited EGF-stimulated growth of transfected murine fibroblasts (3T3) that express the gene for human EGF receptor (Velu, T. J., Beguinot, L., Vass, W. C., Zhang, K., Pastan, I., and Lowy, D. R. (1989) J. Cell. Biochem. 39, 153-166). De-N-acetyl ganglioside GM3 had no effect on the growth of these cells. Growth of control fibroblasts, which lack endogenous EGF receptors (Pruss, R. M., and Herschman, H. R. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 3918-3921), was not affected by the presence of either ganglioside. Similarly, ganglioside GM3, but not de-N-acetyl ganglioside GM3, inhibited the EGF-dependent incorporation of [3H]thymidine into DNA by transfected fibroblasts. Incorporation of labeled thymidine into DNA of control fibroblasts was not affected by the presence of either ganglioside. These studies indicate that ganglioside GM3, but not its deacetylated analogue, can affect EGF receptor kinase activity in intact membranes.(ABSTRACT TRUNCATED AT 400 WORDS)
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División Celular/efectos de los fármacos , Factor de Crecimiento Epidérmico/metabolismo , Receptores ErbB/efectos de los fármacos , Gangliósido G(M3)/análogos & derivados , Gangliósido G(M3)/farmacología , Proteínas Tirosina Quinasas/metabolismo , Animales , Bovinos , ADN/biosíntesis , Receptores ErbB/metabolismo , Humanos , Ratones , Octoxinol , Fosforilación , Polietilenglicoles , Tensoactivos , Transfección , Células Tumorales CultivadasRESUMEN
Binary mixtures of fluid phase phosphatidylethanolamines at pH 10 were treated with the bifunctional cross-linking reagent dimethylsuberimidate. Analysis of the dimeric species formed demonstrated that the phospholipid species in dimyristoylphosphatidylethanolamine/dielaidoylphosphatidylethanolamine mixtures at 52 degrees C and dielaidoylphosphatidylethanolamine/dilauroylphosphatidylethanolamine mixtures at 41 degrees C were randomly arranged. Analysis of the dimeric species formed in dipalmitoylphosphatidylethanolamine/dioleoylphosphatidylethanolamine mixtures at 68 degrees C showed that this mixture was very close to being randomly arranged, with just a slight propensity of like phospholipid species to cluster.
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Dimetil Suberimidato/química , Fluidez de la Membrana , Lípidos de la Membrana , Fosfatidiletanolaminas/química , Reactivos de Enlaces Cruzados , Concentración de Iones de Hidrógeno , TemperaturaRESUMEN
Partitioning of two parinaroyl phosphatidylethanolamines and two parinaroyl phosphatidylglycerols between solid and fluid phase phospholipids was examined. Fluorescence quantum yields and fluorescence polarization measurements were used to calculate Ks/fp, the solid to fluid partition coefficient of each probe (Sklar, L.A., Miljanich, G.P. and Dratz, E.A. (1979) Biochemistry 18, 1707-1716). In the immiscible mixture dipalmitoylphosphatidylcholine and dilinoleylphosphatidylcholine, both 1-palmitoyl-2-trans-parinaroylphosphatidylethanolamine and 1-palmitoyl-2-transparinaroylphosphatidylglycerol partitioned preferentially into solid phase lipid with mean Ks/fp values (calculated from quantum yields) of 3.4 +/- 1.5 and 2.1 +/- 0.7, respectively. In contrast, 1-oleoyl-2-cis-parinaroylphosphatidylethanolamine and 1-oleoyl-2-cis-parinaroylphosphatidylglycerol partitioned preferentially into fluid phase lipid in the same model system with mean Ks/fp values (calculated from quantum yields) of 0.44 +/- 0.26 and 0.16 +/- 0.07, respectively. Fluorescence polarization data on the same four parinaroyl phospholipids in mixtures of solid-phase dimyristoylphosphatidyl ethanolamine and fluid-phase dilinoleoylphosphatidylglycerol were similar to those obtained in the immiscible phosphatidylcholine system, demonstrating that the partitioning of these probes is not strongly dependent on head group. Knowledge of the partition properties of these fluorescent probes is relevant to use of these probes in investigation of the phase behavior of Escherichia coli inner membrane lipids, since phosphatidylethanolamine and phosphatidylglycerol species account for approximately 95% of these lipids.
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Lípidos de la Membrana/análisis , Fosfatidiletanolaminas/análisis , Fosfatidilgliceroles/análisis , Fosfolípidos/análisis , 1,2-Dipalmitoilfosfatidilcolina/análisis , Escherichia coli/análisis , Polarización de Fluorescencia , Lípidos/análisis , Matemática , Modelos Teóricos , Soluciones/análisis , Espectrometría de Fluorescencia , Espectrofotometría , TemperaturaRESUMEN
Dimethylsuberimidate was reacted with aqueous dispersions of dipalmitoylphosphatidylethanolamine, dimyristoylphosphatidylethanolamine, dilauroylphosphatidylethanolamine, and dielaidoylphosphatidylethanolamine at pH 10 and at pH 8. The amount of amidine dimer formation was about four times greater above the gel-to-fluid phase transition of each lipid than below the transition. The transition temperature of each phosphatidylethanolamine, measured by steady-state fluorescence anisotropy of cis-parinaric acid, was lower at pH 10 than at pH 8 or in water. The ability of dimethylsuberimidate to discriminate between phosphatidylethanolamines in the fluid and gel phases should allow use of this reagent to identify phosphatidylethanolamine species within the gel or fluid lipid phase.