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1.
Cell Rep Med ; 2(1): 100164, 2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33521696

RESUMO

Convalescent plasma (CP) is widely used to treat COVID-19, but without formal evidence of efficacy. Here, we report the beneficial effects of CP in a severely ill COVID-19 patient with prolonged pneumonia and advanced chronic lymphocytic leukemia (CLL), who was unable to generate an antiviral antibody response of her own. On day 33 after becoming symptomatic, the patient received CP containing high-titer (ID50 > 5,000) neutralizing antibodies (NAbs), defervesced, and improved clinically within 48 h and was discharged on day 37. Hence, when present in sufficient quantities, NAbs to SARS-CoV-2 have clinical benefit even if administered relatively late in the disease course. However, analysis of additional CP units revealed widely varying NAb titers, with many recipients exhibiting endogenous NAb responses far exceeding those of the administered units. To obtain the full therapeutic benefits of CP immunotherapy, it will thus be important to determine the neutralizing activity in both CP units and transfusion candidates.


Assuntos
COVID-19/terapia , Idoso , Anticorpos Neutralizantes/administração & dosagem , COVID-19/complicações , COVID-19/patologia , COVID-19/virologia , Feminino , Humanos , Imunização Passiva , Hospedeiro Imunocomprometido , Leucemia Linfocítica Crônica de Células B/complicações , Leucemia Linfocítica Crônica de Células B/patologia , Pulmão/diagnóstico por imagem , SARS-CoV-2/isolamento & purificação , Índice de Gravidade de Doença , Tomografia Computadorizada por Raios X , Soroterapia para COVID-19
2.
Curr Med Chem ; 25(28): 3374-3412, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29446723

RESUMO

Inflammatory Bowel Diseases (IBD) comprised of two disorders of idiopathic chronic intestinal inflammation that affect about three million people worldwide: Crohn's disease and ulcerative colitis. Nowadays, the first-line of treatment for patients with mild to moderate symptoms of IBD is comprised of corticosteroids, immunosuppressants, antibiotics, and biological agents. Unfortunately, none of these drugs are curative, and their long-term use may cause severe side effects and complications. Almost 40% of IBD patients use alternative therapies to complement the conventional one, and flavonoids are gaining attention for this purpose. The biological properties of flavonoids are well documented and their antioxidant and anti-inflammatory activities have been arousing attention in the scientific community. Flavonoids are the most widely distributed polyphenols in plants and fruits, making part of the human diet. Taking into account that all ingested flavonoids are expected to exert biological actions at the gastrointestinal level, research on the modulatory effect of these compounds in IBD is of paramount importance. This review intends to summarize, in an integrated and comprehensive form, the effect of flavonoids, both in vitro and in vivo, in the different phases of the characteristic IBD inflammatory network.


Assuntos
Flavonoides/uso terapêutico , Doenças Inflamatórias Intestinais/tratamento farmacológico , Animais , Anti-Inflamatórios/uso terapêutico , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/metabolismo , Colite Ulcerativa/patologia , Doença de Crohn/tratamento farmacológico , Doença de Crohn/metabolismo , Doença de Crohn/patologia , Humanos , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Espécies Reativas de Oxigênio/metabolismo , Fatores de Transcrição/metabolismo
3.
J R Soc Interface ; 14(133)2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28855387

RESUMO

Daunorubicin is extensively used in chemotherapy for diverse types of cancer. Over the years, evidence has suggested that the mechanisms by which daunorubicin causes cytotoxic effects are also associated with interactions at the membrane level. The aim of the present work was to study the interplay between daunorubicin and mimetic membrane models composed of different ratios of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), sphingomyelin (SM) and cholesterol (Chol). Several biophysical parameters were assessed using liposomes as mimetic model membranes. Thereby, the ability of daunorubicin to partition into lipid bilayers, its apparent location within the membrane and its effect on membrane fluidity were investigated. The results showed that daunorubicin has higher affinity for lipid bilayers composed of DMPC, followed by DMPC : SM, DMPC : Chol and lastly by DMPC : SM : Chol. The addition of SM or Chol into DMPC membranes not only increases the complexity of the model membrane but also decreases its fluidity, which, in turn, reduces the amount of anticancer drug that can partition into these mimetic models. Fluorescence quenching studies suggest a broad distribution of the drug across the bilayer thickness, with a preferential location in the phospholipid tails. The gathered data support that daunorubicin permeates all types of membranes to different degrees, interacts with phospholipids through electrostatic and hydrophobic bonds and causes alterations in the biophysical properties of the bilayers, namely in membrane fluidity. In fact, a decrease in membrane fluidity can be observed in the acyl region of the phospholipids. Ultimately, such outcomes can be correlated with daunorubicin's biological action, where membrane structure and lipid composition have an important role. In fact, the results indicate that the intercalation of daunorubicin between the phospholipids can also take place in rigid domains, such as rafts that are known to be involved in different receptor processes, which are important for cellular function.


Assuntos
Permeabilidade da Membrana Celular , Membrana Celular/química , Daunorrubicina/química , Membranas Artificiais , Modelos Químicos , Animais , Membrana Celular/metabolismo , Daunorrubicina/farmacocinética , Humanos , Lipídeos de Membrana/química , Lipídeos de Membrana/metabolismo
4.
Inflammation ; 40(4): 1155-1165, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28405852

RESUMO

Flavonoids are known to react with neutrophil-generated hypochlorous acid (HOCl) at inflammation loci to form stable mono- and dichlorinated products. Some of these products have been shown to retain or even enhance their inflammatory potential, but further information is required in a broader approach to inflammatory mechanisms. In that sense, we performed an integrated evaluation on the anti-inflammatory potential of a panel of novel chlorinated flavonoids and their parent compounds, in several steps of the complex inflammatory cascade, namely, in the activity of cyclooxygenase (COX)-1 and COX-2, and in the production of cytokines [interleukin (IL)-6, IL-1ß, tumor necrosis factor (TNF)], and the chemokine, IL-8, as well as in the production of reactive species, using human whole blood as a representative in vitro model, establishing, whenever possible, a structure-activity relationship. Although luteolin was the most active compound, chlorinated flavonoids demonstrated a remarkable pattern of activity for the resolution of the inflammatory processes. Our results demonstrated that 6-chloro-3',4',5,7-tetrahydroxyflavone deserves scientific attention due to its ability to modulate the reactive species and cytokines/chemokine production. In this regard, the therapeutic potential of flavonoids' metabolites, and in this particular case the chlorinated flavonoids, should not be neglected.


Assuntos
Flavonoides/farmacologia , Ácido Hipocloroso/química , Mediadores da Inflamação/farmacologia , Inflamação/sangue , Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Citocinas/biossíntese , Citocinas/metabolismo , Flavonoides/química , Humanos , Hidrocarbonetos Clorados , Mediadores da Inflamação/química , Interleucina-8/metabolismo , Relação Estrutura-Atividade
5.
Biochim Biophys Acta Biomembr ; 1859(5): 941-948, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28153496

RESUMO

The present work aimed to study the interactions between the anticancer drug daunorubicin and lipid membrane mimetic models of cancer cells composed by their most representative classes of phospholipids, with different degrees of complexity. Regarding these anticancer drug-membrane interactions, several biophysical parameters were assessed using liposomes (LUVs) composed of different molar ratios of DMPC, DOPC, DPPS, DOPE and Chol. In this context, daunorubicin's membrane concentration was determined by calculating its partition coefficient (Kp) between liposomes and water using derivative UV/vis spectrophotometry at 37°C and pH6.3, a typical tumoral microenvironment. Characterization of the zeta potential of such model membranes, in both the absence and presence of the compound, was accomplished through Electrophoretic Light Scattering (ELS). Fluorescence quenching studies, which determine the location of the drug within the bilayer, were carried out using liposomes labelled with DPH and TMA-DPH, fluorescent probes with known membrane position. Temperature dependent steady-state anisotropy assays were also performed to measure the daunorubicin effect on the membranes' microviscosity. The overall results support that daunorubicin permeation depends on the phospholipid membrane composition and causes alterations in the biophysical properties of the bilayers, namely in the membrane fluidity. The interaction of daunorubicin with the studied phospholipids is mainly driven by electrostatic and hydrophobic interactions. These insights demonstrated that not only membranes can affect daunorubicin accumulation in cells but the compound can alter the properties of membranes. The changes produced by daunorubicin on the lipid structure may constitute an additional mechanism of action, which might lead to modifications in the location and, consequently, the activity of membrane signaling proteins.


Assuntos
Antibióticos Antineoplásicos/farmacologia , Daunorrubicina/farmacologia , Bicamadas Lipídicas/química , Anisotropia , Biofísica , Daunorrubicina/farmacocinética , Corantes Fluorescentes , Concentração de Íons de Hidrogênio , Luz , Fluidez de Membrana/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Espalhamento de Radiação
6.
Food Funct ; 7(5): 2249-57, 2016 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-27142444

RESUMO

Solanum diploconos is an unexploited Brazilian native fruit that belongs to the same genus of important food crops, such as tomato (Solanum lycorpersicum) and potato (Solanum tuberosum). In this study, we determined, for the first time, the profile of bioactive compounds (phenolic compounds, carotenoids, ascorbic acid and tocopherols) of the freeze-dried pulp and peel of Solanum diploconos fruits, as well as of an extract obtained from the whole fruit. Additionally, the antioxidant potential of the whole fruit extract was evaluated in vitro, against reactive oxygen species (ROS) and reactive nitrogen species (RNS). Eighteen phenolic compounds were identified in the peel and pulp and 6 compounds were found in the whole fruit extract. Coumaric, ferulic and caffeic acid derivatives were revealed to be the major phenolic constituents. All-trans-ß-carotene was the major carotenoid (17-38 µg g(-1), dry basis), but all-trans-lutein and 9-cis-ß-carotene were also identified. The peel and pulp presented <2 µg per mL of tocopherols, and ascorbic acid was not detected. The whole fruit extract exhibited scavenging capacity against all tested ROS and RNS (IC50 = 14-461 µg mL(-1)) with high antioxidant efficiency against HOCl. Thus, Solanum diploconos fruits may be seen as a promising source of bioactive compounds with high antioxidant potential against the most physiologically relevant ROS and RNS.


Assuntos
Antioxidantes/análise , Frutas/química , Extratos Vegetais/química , Solanum/química , Antioxidantes/química , Ácido Ascórbico/análise , Ácido Ascórbico/genética , Brasil , Ácidos Cafeicos/análise , Ácidos Cafeicos/química , Carotenoides/análise , Carotenoides/química , Cromatografia Líquida de Alta Pressão , Ácidos Cumáricos/análise , Ácidos Cumáricos/química , Sequestradores de Radicais Livres/análise , Sequestradores de Radicais Livres/química , Peróxido de Hidrogênio , Ácido Hipocloroso , Luteína/análise , Luteína/química , Óxido Nítrico , Ácido Peroxinitroso , Fenóis/análise , Fenóis/química , Espécies Reativas de Nitrogênio/química , Espécies Reativas de Oxigênio/química , Oxigênio Singlete , Superóxidos , Tocoferóis/análise , Tocoferóis/química
7.
Med Res Rev ; 35(5): 877-936, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25926332

RESUMO

Inflammation is a natural, carefully orchestrated response of the organism to tissue damage, involving various signaling systems and the recruitment of inflammatory cells. These cells are stimulated to release a myriad of mediators that amplify the inflammatory response and recruit additional cells. These mediators present numerous redundancies of functions, allowing a broad and effective inflammatory response, but simultaneously make the understanding of inflammation pathways much difficult. The extent of the inflammatory response is usually self-limited, although it depends on the balance between the pro- and anti-inflammatory signals. When that equilibrium is dislocated, a more widespread inflammatory response may take place. Flavonoids have been shown to be possible alternatives to the traditionally molecules used as anti-inflammatory agents. In fact, the biological activities of flavonoids include the modulation of the diverse phases of inflammatory processes, from the gene transcription and expression to the inhibition of the enzymatic activities and the scavenging of the reactive species. In the present review, the inflammatory network is widely revised and the flavonoids' broad spectrum of action in many of the analyzed inflammatory pathways is revised. This kind of integrated revision is original in the field, providing the reader the simultaneous comprehension of the inflammatory process and the potential beneficial activities of flavonoids.


Assuntos
Flavonoides/química , Inflamação/fisiopatologia , Animais , Basófilos/citologia , Plaquetas/citologia , Células Dendríticas/citologia , Eicosanoides/química , Células Endoteliais/citologia , Eosinófilos/citologia , Células Epiteliais/citologia , Fibroblastos/citologia , Granulócitos/citologia , Humanos , Lipídeos/química , Linfócitos/citologia , Macrófagos/citologia , Mastócitos/citologia , Monócitos/citologia , Neutrófilos/citologia , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo
8.
Med Chem ; 11(6): 560-6, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25665653

RESUMO

Nuclear factor kappa B (NF-kB) is one of the most important transcription factors whose modulation triggers a cascade of signaling events, namely the expression of many cytokines, enzymes, chemokines, and adhesion molecules, some of which being potential key targets for intervention in the treatment of inflammatory conditions. The 2-styrylchromones (2-SC) designation represents a well-recognized group of natural and synthetic chromones, vinylogues of flavones (2-phenylchromones). Several 2-SC were recently tested for their anti-inflammatory potential, regarding the arachidonic acid metabolic cascade, showing some motivating results. In addition, several flavones with structural similarities to 2-SC have shown NF-kB inhibitory properties. Hence, the aim of the present work was to continue the investigation on the interference of 2-SC in inflammatory pathways. Herein we report their effects on lipopolysaccharide (LPS)-induced NF-kB activation and consequent production of proinflammatory cytokines/chemokine, using a human monocytic cell line (THP-1). From the twelve 2-SC tested, three of them were able to significantly inhibit the NF-kB activation and to reduce the production of the proinflammatory cytokines/chemokine. The compound 3',4',5-trihydroxy-2- styrylchromone stood up as the most active in both assays, being a promising candidate for an anti-inflammatory drug.


Assuntos
Cromonas/farmacologia , Citocinas/biossíntese , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , NF-kappa B/antagonistas & inibidores , Sobrevivência Celular , Cromonas/síntese química , Cromonas/química , Relação Dose-Resposta a Droga , Humanos , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Estrutura Molecular , NF-kappa B/metabolismo , Relação Estrutura-Atividade , Células Tumorais Cultivadas
9.
Biochim Biophys Acta ; 1828(2): 896-905, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23022131

RESUMO

This work focuses on the interaction of mycolic acids (MAs) and two antimycobacterial compounds (Rifabutin and N'-acetyl-Rifabutin) at the pulmonary membrane level to convey a biophysical perspective of their role in disease. For this purpose, accurate biophysical techniques (Langmuir isotherms, Brewster angle microscopy, and polarization-modulation infrared reflection spectroscopy) and lipid model systems were used to mimic biomembranes: MAs mimic bacterial lipids of the Mycobacterium tuberculosis (MTb) membrane, whereas Curosurf® was used as the human pulmonary surfactant (PS) membrane model. The results obtained show that high quantities of MAs are responsible for significant changes on PS biophysical properties. At the dynamic inspiratory surface tension, high amounts of MAs decrease the order of the lipid monolayer, which appears to be a concentration dependent effect. These results suggest that the amount of MAs might play a critical role in the initial access of the bacteria to their targets. Both molecules also interact with the PS monolayer at the dynamic inspiratory surface. However, in the presence of higher amounts of MAs, both compounds improve the phospholipid packing and, therefore, the order of the lipid surfactant monolayer. In summary, this work discloses the putative protective effects of antimycobacterial compounds against the MAs induced biophysical impairment of PS lipid monolayers. These protective effects are most of the times overlooked, but can constitute an additional therapeutic value in the treatment of pulmonary tuberculosis (Tb) and may provide significant insights for the design of new and more efficient anti-Tb drugs based on their behavior as membrane ordering agents.


Assuntos
Antibacterianos/farmacologia , Biofísica/métodos , Membrana Celular/química , Ácidos Micólicos/química , Surfactantes Pulmonares/química , 1,2-Dipalmitoilfosfatidilcolina/química , Antituberculosos/farmacologia , Produtos Biológicos/farmacologia , Células Epiteliais/citologia , Humanos , Lipídeos/química , Modelos Químicos , Mycobacterium tuberculosis/metabolismo , Fosfatidilserinas/química , Fosfolipídeos/farmacologia , Alvéolos Pulmonares/citologia , Rifabutina/farmacologia , Espectrofotometria/métodos , Propriedades de Superfície , Tensoativos/química
10.
Talanta ; 84(5): 1314-7, 2011 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-21641444

RESUMO

A fully automated multipumping flow system (MPFS) using water-soluble CdTe quantum dots (QD) as sensitizers is proposed for the chemiluminometric determination of the anti-diabetic drugs gliclazide and glipizide in pharmaceutical formulations. The nanocrystals acted as enhancers of the weak CL emission produced upon oxidation of sulphite by Ce(IV) in acidic medium, thus improving sensitivity and expanding the dynamical analytical concentration range. By interacting with the QD, the two analytes prevented their sensitizing effect yielding a chemiluminescence quenching of the Ce(IV)-SO(3)(2-)CdTe QD system. The pulsed flow inherent to MPFS assured a fast and efficient mixing of all solutions inside the flow cell, circumventing the need for a reaction coil and facilitating the monitoring of the short-lived generated chemiluminescent species. QD crystal size, concentration and spectral region for measurement were investigated.

11.
Luminescence ; 26(6): 571-8, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21308949

RESUMO

In this work, an automated flow-based procedure for the screening of the effect of the different phenolic compounds on the chemiluminescence (CL) luminol-hydrogen peroxide-horseradish peroxidase (HRP) system is presented. This procedure involves the combination of multisyringe flow injection analysis (MFSIA) and sequential injection analysis (SIA) techniques and exploits the ability of the different subgroups of phenols, such as cholorophenols, nitrophenols, methylphenols and polyphenols, to enhance or inhibit the described CL system. The implementation of this reaction in the SIA-MSFIA system enabled favourable and precise conditions to evaluate the effect of phenolic compounds, as it involves an in-line reaction between the phenolic derivative, hydrogen peroxide and peroxidase and subsequent oxidized HRP intermediates generation prior to the fast reaction with the chemiluminogenic reagent. Several studies were then performed with the aim of establishing the appropriate flow system configuration and reaction conditions. It was shown that phenol and chlorophenols produce an enhanced CL response and nitrophenols, methylphenols and polyphenols are inhibitors within the range of concentrations studied (1-100 mg/L). Based on these studies, the developed method was applied to the determination of total polyphenol and phenol content in wine/grape seeds and water samples, respectively, and the results obtained showed good agreement with those furnished by the corresponding Folin-Ciocalteu and 4-aminoantipyrine reference methods. The developed approach is further pursued by designing an automated generic tool for performing studies of peroxidase-catalysed CL reactions of luminol focused on the detection of compounds that will affect the rate of those reactions.


Assuntos
Peroxidase do Rábano Silvestre/química , Peróxido de Hidrogênio/química , Luminol/química , Fenóis/química , Luminescência
12.
Anal Bioanal Chem ; 397(7): 3005-14, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20563794

RESUMO

In the present work, a fluorimetric automatic method based on multisyringe flow injection analysis (MSFIA) was developed for in vitro evaluation of scavenging capacity against nitric oxide (NO) using 4,5-diaminofluorescein (DAF-2) as an NO-selective fluorogenic probe. The MSFIA manifold was assembled to perform the in-line generation of NO and the competitive reaction of putative scavenger molecules and DAF-2 with NO at conditions close to those found in vivo regarding temperature (37 degrees C), pH (7.4), and concentration of NO (less than 1 microM). This approach allowed the evaluation of scavenging capacity against NO by endogenous antioxidant molecules, pharmaceutical compounds, and human plasma. IC(50) values were calculated for rutin (1.30 +/- 0.02 microM, positive control), cysteine (321 +/- 8 microM), reduced glutathione (1106 +/- 93 microM), uric acid (134 +/- 12 microM), dipyrone (1.36 +/- 0.06 microM), and captopril (363 +/- 28 microM). A high degree of automation was attained as the successive dilution of antioxidant standard solutions required for IC(50) assessment was performed automatically, in a dilution chamber placed in the flow system. A determination throughput of 16 h(-1) and a good precision were attained (relative standard deviation between 1.6 and 9.0%), fostering the application of the proposed method to routine/screening analysis of scavenging capacity against NO.


Assuntos
Análise de Injeção de Fluxo/métodos , Sequestradores de Radicais Livres/análise , Óxido Nítrico/análise , Automação , Análise de Injeção de Fluxo/instrumentação , Sequestradores de Radicais Livres/sangue , Humanos
13.
Talanta ; 81(4-5): 1840-6, 2010 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-20441983

RESUMO

In the present work, an automatic method based on multisyringe flow injection analysis (MSFIA) was developed for determination of scavenging capacity against H(2)O(2) using a non-enzymatic fluorimetric assay for H(2)O(2) detection based on the formation of europium-tetracycline-H(2)O(2) complex. The MSFIA-fluorimetric methodology was developed to perform in-line the scavenging reaction of H(2)O(2) prior to reaction of the remaining H(2)O(2) with the fluorescent probe, using conditions close to those found in vivo regarding pH (6.9), temperature (37 degrees C) and H(2)O(2) concentration (25microM). This approach allowed the evaluation of scavenging capacity against H(2)O(2) in a non-competitive (antioxidant+H(2)O(2)) or a competitive (antioxidant+H(2)O(2)+biological target) scheme. Using the first strategy, IC(50) values determined for the antioxidant compounds glutathione reduced (1191+/-46microM) and pyruvate (446+/-49microM) were lower than those obtained for biological targets such as cysteine (2616+/-182microM), taurine (359+/-38mM) and adenine (2224+/-214microM), indicating that reactivity towards H(2)O(2) was higher for antioxidant compounds than for biological targets. However, when a competitive scheme was applied, the scavenging effectiveness against H(2)O(2) depended on the biological molecule present, showing that antioxidant assessment should also take into consideration the concomitant reactivity of biological molecules or structures that are prone to oxidative damage.


Assuntos
Antioxidantes/química , Fluorometria/métodos , Peróxido de Hidrogênio/química , Adenina/química , Técnicas de Química Analítica , Cisteína/química , Desenho de Equipamento , Európio/química , Concentração Inibidora 50 , Limite de Detecção , Estresse Oxidativo , Software , Taurina/química , Tetraciclina/química
14.
Mini Rev Med Chem ; 10(1): 1-7, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20380638

RESUMO

2-Styrylchromones (2-SC) are chromone derivatives characterized by the attachment of a styryl group to the 2-position of the chromone structure. The 2-styrylchromone structure has been demonstrated to bear important biological activities such as antiallergic, antitumor, affinity and selectivity for A(3) adenosine receptors, antiviral, antioxidant and anti-inflammatory. The aim of the present paper was to review the available information on this field.


Assuntos
Cromonas/farmacologia , Estirenos/farmacologia , Animais , Antialérgicos/química , Antialérgicos/metabolismo , Antialérgicos/farmacologia , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/metabolismo , Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Antioxidantes/química , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Antivirais/química , Antivirais/metabolismo , Antivirais/farmacologia , Cromonas/química , Cromonas/metabolismo , Humanos , Receptor A3 de Adenosina/metabolismo , Rhinovirus/efeitos dos fármacos , Relação Estrutura-Atividade , Estirenos/química , Estirenos/metabolismo
15.
Methods Mol Biol ; 606: 167-88, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20013397

RESUMO

Cellular membranes, which contain abundant phospholipids, such as phosphatidylcholine, are major targets subjected to the damage caused by free radicals. Cellular damage due to lipid oxidation is strongly associated with ageing, carcinogenesis and other diseases. In addition, lipid oxidation is an important deteriorative reaction in the processing and storage of lipid-containing foods. Liposomes have been used extensively as biological models for in vitro lipid oxidation studies. The resemblance between the liposomal and membrane bilayer core makes liposomes a very useful tool to investigate the significance of the antioxidant-membrane interactions for antioxidant activity. The antioxidant activity of a compound is strongly influenced by numerous factors including the nature of the lipid substrate, the hydrophilic-lipophilic balance of the antioxidant, the physical and chemical environments of the lipids, and various other interfacial interactions. Thus, compounds that are effective antioxidants in one model system or food matrix may be unsuitable in other systems.This chapter describes fluorescent probes-based methods commonly used for testing antioxidant activity in liposomes and stresses the need to combine antioxidant assays and drug-membrane interaction studies to get a better description of the antioxidants' profile considering their location in lipid bilayer and their effect on membrane fluidity and consequently provide additional information to that obtained currently from assays performed in aqueous buffer media.


Assuntos
Antioxidantes/metabolismo , Antioxidantes/farmacologia , Corantes Fluorescentes/análise , Lipossomos/metabolismo , Antioxidantes/análise , Radicais Livres/metabolismo , Bicamadas Lipídicas/metabolismo , Fluidez de Membrana/efeitos dos fármacos
16.
Talanta ; 79(4): 1169-76, 2009 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-19615527

RESUMO

The determination of hydrogen peroxide (H(2)O(2)) and the evaluation of scavenging capacity against this species were performed using five colorimetric reactions, which were adapted to flow injection analysis. The reactions chosen were based on the oxidation of iodide (I(-) method), on the formation of titanium-peroxide complex (TiP method), on the formation of titanium-xylenol orange-peroxide complex (TiXoP method), on the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB method) and on the co-oxidation of phenol-4-sulfonic acid and 4-aminoantipyrine (PSA/4-AAP method). The operational conditions were studied in order to improve the sensitivity of each method. Concerning to the method sensitivity, the ranking order was TMB method>I(-) method>TiXoP method approximately PSA/4-AAP method>TiP method. All methods showed an excellent repeatability (RSD<2%) and, except for I(-) method, relative deviations from the reference method were <1.9%. The FIA manifolds were adapted to perform the determination of scavenging capacity against H(2)O(2) and glutathione (GSH) was applied as model compound. TiP and TiXoP methods were not suitable as no inhibition or an increase of analytical signal was attained. PSA/4-AAP method was chosen for further application to dietary phenolics and pharmaceutical compounds, providing IC(50) values for those compounds that are fast reacting antioxidants.

17.
J Med Food ; 12(1): 175-83, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19298212

RESUMO

Eucalyptus globulus Labill. (Family Myrtaceae) is a plant of Australian origin, with a reported therapeutic use in airway inflammatory diseases. Considering that reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been implicated in the pathogenesis of airway inflammatory diseases such as asthma and chronic obstructive pulmonary disease, an effective scavenging activity against these reactive species may contribute for the therapeutic effect of this plant. In the present study, a water extract of E. globulus leaves was evaluated for its putative in vitro scavenging effects on ROS (HO(*), O(2)(*-), ROO(*), and H(2)O(2)) and RNS ((*)NO and ONOO(-)) and on 2,2-diphenyl-1-picrylhydrazyl radical (DPPH). Qualitative and quantitative analyses of the extract's phenolic composition were also performed. The Eucalyptus leaf water extract presented a remarkable capacity to scavenge all the reactive species tested, with all the 50% inhibitory concentrations being found at the mug/mL level. Phytochemical analysis showed the presence of polyphenols such as flavonoids (rutin and quercitrin) and phenolic acids (chlorogenic acid and ellagic acid), which may be partially responsible for the observed antioxidant activity. These observations provide further support, beyond the well-known antibacterial and antiviral activities of the Eucalyptus plant, for its reported use in traditional medicine such as in the treatment of airway inflammatory diseases, considering the important role of ROS and RNS in the inflammatory process, although further studies are needed to prove the bioavailability of the antioxidants/antibacterial compounds of the extract as well as the ability of the active compounds to reach specific tissues and to act in them.


Assuntos
Eucalyptus/química , Sequestradores de Radicais Livres/farmacologia , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Compostos de Bifenilo , Sequestradores de Radicais Livres/isolamento & purificação , Fenóis/análise , Fenóis/farmacologia , Picratos , Folhas de Planta
18.
Talanta ; 77(5): 1559-66, 2009 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-19159764

RESUMO

The role and importance of antioxidants in different fields, ranging from physiology to food technology, have become evident in the past years, requiring adequate analytical methodologies. Therefore, the determination of antioxidant capacity as a routine or screening analysis fosters its automation. In this context, several flow injection methods based on scavenging of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonate) radical cation (ABTS(+)) or 2,2-diphenyl-1-picrylhydrazyl radical (DDPH) or based on the determination of total reducing capacity have been proposed. The objective of the present review is to critically compare the different approaches, regarding their degree of automation, their performance vs. the respective batch procedure and its applicability to real samples.


Assuntos
Antioxidantes/análise , Análise de Injeção de Fluxo/métodos , Benzotiazóis , Sequestradores de Radicais Livres , Métodos , Fenetilaminas , Ácidos Sulfônicos
19.
Talanta ; 74(5): 1511-9, 2008 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-18371811

RESUMO

This work reports the development of a simple, robust, automated sequential injection analysis (SIA) system for the enzymatic determination of total (tGSH) and oxidized (GSSG) glutathione in human whole blood. The reduced (GSH) glutathione concentration is then obtained as the difference between the tGSH and GSSG concentrations. The determination was based on the DTNB-GSSG reductase recycling assay, which couples the specificity of the GSSG reductase (GR) with an amplification of the response to glutathione, followed by spectrophotometric detection of the 2-nitro-5-thiobenzoic acid (TNB) formed (lambda=412 nm). The implementation of this reaction in a SIA flow system with an in-line dilution strategy permitted the necessary distinct application ranges for tGSH and for GSSG. It also guaranteed the exact timing of fluidic manipulations and precise control of the reaction conditions. The influence of parameters such as reagents concentration, temperature, pH, flow rate of the carrier buffer solution, as well as reaction coil length, etc., on the sensitivity and performance of the SIA system were studied and the optimum reaction conditions subsequently selected. Linear calibration plots were obtained for GSH and GSSG concentrations up to 3.00 and 1.50 microM, with detection limits of 0.031 and 0.014 microM, respectively. The developed methodology showed good precision, with a relative standard deviation (R.S.D.)<5.0% (n=10) for determination of both glutathione forms. Statistical evaluation showed good compliance, for a 95% confidence level, between the results obtained with the SIA system and those furnished by the comparison batch procedure.


Assuntos
Análise de Injeção de Fluxo/métodos , Dissulfeto de Glutationa/sangue , Glutationa/sangue , Análise de Injeção de Fluxo/normas , Humanos , Nitrobenzoatos , Reprodutibilidade dos Testes , Espectrofotometria/métodos , Espectrofotometria/normas , Compostos de Sulfidrila
20.
J Pharm Sci ; 97(8): 3195-206, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17990311

RESUMO

In this work, fluorescence measurements were performed using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH) to evaluate the effects of the interaction of nonsteroidal anti-inflammatory drugs--NSAIDs (meloxicam, lornoxicam, and nimesulide) with several membrane systems (liposomes with and without cholesterol, mouse splenocytes, mouse macrophages cell line--J774, human leukemia monocyte cell line--THP-1, and human granulocytes and mononuclear cells). DPH fluorescence quenching studies revealed that the NSAIDs studied were able to efficiently quench the probe located in membrane hydrocarbon region. Fluorescence anisotropy measurements were also made to investigate the effects on membrane fluidity resulting from the interaction between the drugs and membrane systems. All the anti-inflammatory drugs studied show an increase in the membrane fluidity in a concentration dependent manner. Results obtained provide an insight into NSAIDs capacity to be inserted in lipid bilayers and alter the lipid dynamics. The induced changes in lipid dynamics may modulate the activity of inflammatory enzymes or may be related with deleterious topical action of NSAIDs on gastric phospholipid fluidity.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Fluidez de Membrana/efeitos dos fármacos , Animais , Linhagem Celular , Polarização de Fluorescência , Corantes Fluorescentes , Humanos , Bicamadas Lipídicas , Lipossomos , Camundongos , Camundongos Endogâmicos BALB C , Sulfonamidas/farmacologia
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