Amiodarone interactions with membrane lipids and with growth of Bacillus stearothermophilus used as a model

Abstract The thermophilic eubacterium Bacillus stearothermophilus was used as a model to study the effects of amiodarone (2-butyl-3-[3',5'diido-4'a-diethyl-aminoethoxybenzoyl]-benzofuran) in lipid organization and in bacterial growth. Effects on the structural order of lipids were assessed by fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH), probing the bilayer core, and of the propionic acid derivative 3-[p-(6-phenyl)-1,3,5-hexatrienyl] phenylpropionic acid (DPH-PA), probing the outer regions of the bilayer. Amiodarone fluidizes bacterial polar lipid bilayers for temperatures below the phase transition midpoint, and orders the fluid phase of the bacterial polar lipids, as evaluated by DPH and DPH-PA. The ordering and disordering effects, which are concentration dependent, are more extensive when detected by DPH relative to DPH-PA. Growth studies performed in parallel revealed that amiodarone inhibits bacterial growth as a function of concentration. Amiodarone concentrations in the range from 1 to 2.5 ?M increased the lag time, decreased the specific growth rate, and decreased the final cell density. Furthermore, 3 ?M amiodarone completely inhibited growth. These in vivo effects of amiodarone can be related to its ability to perturb the phospholipid bilayer structure, whose integrity is essential for cell function, viability, and growth

Toxicity assessment of tamoxifen by means of a bacterial model

Abstract A strain of Bacillus stearothermophilus was used as a model to study physical perturbations induced in the membrane by the cytostatic tamoxifen (TAM). This study was carried out using two lines of criteria: (1) bacterial growth, and temperature growth range, with determination of growth parameters as a function of TAM concentration; and (2) biophysical studies by differential scanning calorimetry (DSC) and by means of two fluorescent probes to evaluate perturbations promoted by the drug on the structural order of bacterial lipid membranes. The inhibition of growth induced by TAM, the structural bilayer disordering, and the shift in the phase transition temperature to a lower range were also determined in the presence of Ca2+, i.e., a natural membrane stabilizer, to elucidate further perturbing effects of TAM on membranes with putative implications in cell toxicity. Growth inhibition promoted by TAM is potentiated by an increase in growth temperature above the optimal range, but attenuated or relieved by the addition of 2.5 mM Ca2+ to the culture medium. Consistently, fluorescence polarization and DSC studies showed that Ca2+ ions (2.5 mM) effectively compensated for the destabilizing effects promoted by TAM in bacterial lipid membranes

Mitochondrial bioenergetics is affected by the herbicide paraquat

AbstractThe potential toxicity of the herbicide paraquat (1,1-dimethyl-4,4′-bipyridylium dichloride) was tested in bioenergetic functions of isolated rat liver mitochondria. Paraquat increases the rate of State 4 respiration, doubling at 10 mM, indicating uncoupling effects. Additionally, State 3 respiration is depressed by about 15%, at 10 mM paraquat, whereas uncoupled respiration in the presence of CCCP is depressed by about 30%. Furthermore, paraquat partially inhibits the ATPase activity through a direct effect on this enzyme complex. However, at high concentrations (5–10 mM), the ATPase activity is stimulated, probably as consequence of the described uncoupling effect. Depression of respiratory activity is mediated through partial inhibitions of mitochondrial complexes III and IV. Paraquat depresses Δψ as a function of herbicide concentration. In addition, the depolarization induced by ADP is decreased and repolarization is biphasic suggesting a double effect. Repolarization resumes at a level consistently higher than the initial level before ADP addition, for paraquat concentrations up to 10 mM. This particular effect is clear at 1 mM paraquat and tends to fade out with increasing concentrations of the herbicide

Na oficina do dramaturgo : edição crítica de uma obra inédita de Marcelino Mesquita

Esta dissertação de mestrado está dividida em duas partes principais, pelas quais se pretendeu valorizar um dramaturgo português de finais do século XIX: Marcelino Mesquita. A primeira parte deste trabalho insere-o no contexto histórico-literário, faz um esboço de reconstituição da sua fortuna crítica, fala-nos um pouco da sua vida e essencialmente da obra, revelando as opiniões dos poucos que acham ser ele merecedor de alguma referência, elogiosa ou menos abonatória. Da segunda parte, a principal, consta o texto crítico de um inédito seu sem título e não datado, encontrado no seu espólio por um estudioso do autor, cuja divulgação consideramos de grande premência literária. Com efeito, impõem-na a qualidade da peça, a sua modernidade e até uma certa atipicidade em relação à marca distintiva dos seus textos dramáticos. /ABSTRACT - This dissertation is divided into two main parts, and its purpose is to stress the value of a portuguese playwright of the end of the 19th century: Marcelino Mesquita. The first part of this essay introduces the author in his historical and literary time and gives some information about his life and, essentially, about his work. This part also reveals the opinions of the few who consider him worth of some critical reference, both in flattering terms and not so flattering ones... The second part, the most important one, includes the critical text of an unpublished play, with no title nor date. This play was found in the remains of the author by a specialist of his writings and we considered very important, in literary terms, to make it public. In truth, the quality of the play, its modernity and the fact of being atypical deserve this acknowledgement

Changes induced by malathion, methylparathion and parathion on membrane lipid physicochemical properties correlate with their toxicity

Perturbations induced by malathion, methylparathion and parathion on the physicochemical properties of dipalmitoylphosphatidylcholine (DPPC) were studied by fluorescence anisotropy of DPH and DPH-PA and by differential scanning calorimetry (DSC). Methylparathion and parathion (50 [mu]M) increased the fluorescence anisotropy evaluated by DPH-PA and DPH, either in gel or in the fluid phase of DPPC bilayers, but mainly in the fluid phase. Parathion is more effective than methylparathion. On the other hand, malathion had almost no effect. All the three xenobiotics displaced the phase transition midpoint to lower temperature values and broadened the phase transition profile of DPPC, the effectiveness following the sequence: parathion>methylparathion>>malathion. A shifting and broadening of the phase transition was also observed by DSC. Furthermore, at methylparathion/lipid molar ratio of 1/2 and at parathion/lipid molar ratio of 1/7, the DSC thermograms displayed a shoulder in the main peak, in the low temperature side, suggesting coexistence of phases. For higher ratios, the phase transition profile becomes sharp as the control transition, but the midpoint is shifted to the previous shoulder position. Conversely to methylparathion and parathion, malathion did not promote phase separation. The overall data from fluorescence anisotropy and calorimetry indicate that the degree of effect of the insecticides on the physicochemical membrane properties correlates with toxicity to mammals. Therefore, the in vivo effects of organophosphorus compounds may be in part related with their ability to perturb the phospholipid bilayer structure, whose integrity is essential for normal cell function.http://www.sciencedirect.com/science/article/B6T1T-42NY32W-K/1/9c5c8320a8dff42bbf122281b5056b8

Duas peças do espólio de Marcelino Mesquita

Esta obra edita pela primeira vez em volume duas peças pertencentes ao espólio de Marcelino Mesquita

Lipid composition and dynamics of cell membranes of Bacillus stearothermophilus adapted to amiodarone

Bacillus stearothermophilus, a useful model to evaluate membrane interactions of lipophilic drugs, adapts to the presence of amiodarone in the growth medium. Drug concentrations in the range of 1-2 [mu]M depress growth and 3 [mu]M completely suppresses growth. Adaptation to the presence of amiodarone is reflected in lipid composition changes either in the phospholipid classes or in the acyl chain moieties. Significant changes are observed at 2 [mu]M and expressed by a decrease of phosphatidylethanolamine (relative decrease of 23.3%) and phosphatidylglycerol (17.9%) and by the increase of phosphoglycolipid (162%). The changes in phospholipid acyl chains are expressed by a decrease of straight-chain saturated fatty acids (relative decrease of 12.2%) and anteiso-acids (22%) with a parallel increase of the iso-acids (9.8%). Consequently, the ratio straight-chain/branched iso-chain fatty acids decreases from 0.38 (control cultures) to 0.30 (cultures adapted to 2 [mu]M amiodarone). The physical consequences of the lipid composition changes induced by the drug were studied by fluorescence polarization of diphenylhexatriene and diphenylhexatriene-propionic acid, and by differential scanning calorimetry. The thermotropic profiles of polar lipid dispersions of amiodarone-adapted cells are more similar to control cultures (without amiodarone) than those resulting from a direct interaction of the drug with lipids, i.e., when amiodarone was added directly to liposome suspensions. It is suggested that lipid composition changes promoted by amiodarone occur as adaptations to drug tolerance, providing the membrane with physico-chemical properties compatible with membrane function, counteracting the effects of the drug.http://www.sciencedirect.com/science/article/B6VNN-419BF60-K/1/8f2d4fae7f9c131d26230cf4123da94

Connecting people and strengthening inter-institutional cooperation: the case of the Portuguese Sustainable Campus Network (RCS)

Trabalho apresentado em 29th International Sustainable Development Research Society Conference (ISDRS), 2023, Kuala Lumpur, MalasiaN/

Size dependence of the translational diffusion of large integral membrane proteins in liquid-crystalline phase lipid bilayers. A study using fluorescence recovery after photobleaching

The translational diffusion of bovine rhodopsin, the Caz+-activated adenosinetriphosphatase of rabbit muscle sarcoplasmic reticulum, and the acetylcholine receptor monomer of Torpedo marmorata has been examined at a high dilution (molar ratios of lipid/protein 1 3000/1) in liquidcrystalline phase phospholipid bilayer membranes by using the fluorescence recovery after photobleaching technique. These integral membrane proteins having molecular weights of about 37 000 for rhodopsin, about 100000 for the adenosinetriphosphatase, and about 250 000 for the acetylcholine receptor were reconstituted into membranes of dimyristoylphosphatidylcholine (rhodopsin and acetylcholine receptor), soybean lipids (acetylcholine receptor), and a total lipid extract of rabbit muscle sarcoplasmic reticulum (adenosinetriphosphatase). The translational diffusion coefficients of all the proteins at 310 K were found to be in the range (1-3) X cm2/s. In consideration of the sizes of the membranebound portions of these proteins, this result is in agreement with the weak dependence of the translational diffusion coefficient upon diffusing particle size predicted by continuum fluid hydrodynamic models for the diffusion in membranes [Saffman, P. G., & Delbriick, M. (1975) Proc. Natl. Acad. Sci. U.S.A. 72, 3 1 1 1-3 1 131. Lipid diffusion was also examined in the same lipid bilayers with the fluorescent lipid derivative N-(7-nitro-2,1,3-benzoxadiazol-4-yl)dimyristoylphosphatidylethanolamine. The translational diffusion coefficient for this lipid derivative was found to be in the range (9-14) X cm2/s at 310 K. In consideration of the dimensions of the lipid molecule, this value for the lipid diffusion coefficient is in agreement with the continuum fluid hydrodynamic model only if a near-complete slip boundary condition is assumed at the bilayer midplane. Alternatively, kinetic diffusion models [Trauble, H., & Sackmann, E. (1972) J. Am. Chem. SOC9. 4,4499-45101 may have to be invoked to explain the lipid diffusion behavior