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1.
Luminescence ; 38(3): 341-349, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36760184

RESUMO

Chemiluminescence (CL) reactions are widely used for the detection and quantification of many types of analytes. Laccase has previously been proposed in CL reactions; however, its light emission behaviour has not been characterized. This study was conducted to characterize the laccase-luminol system, determine its kinetic parameters, and analyze the effects of protein and OH- concentration on the CL signal. Laccase from Coriolopsis gallica was combined with different concentrations of luminol (125 nM to 4 mM), and the enzyme kinetics were evaluated using diverse kinetic models. The laccase-luminol system was able to produce CL without an intermediate molecule, but it exhibited substrate-inhibition behaviour. A two-site random model was used and suggested that when the first luminol molecule was bound to the active site, laccase affinity for the second luminol molecule was increased. This inhibition effect could be avoided using a low luminol concentration. At 5 µM luminol concentration, 1 mg/ml (0.13 U) laccase is needed to achieve nearly 90% of the maximum CL signal, suggesting that the available luminol could not bind to all active sites. Furthermore, the concentration of NaOH negatively affected the CL signal. The laccase-luminol system represents an alternative to existing CL systems, with potential uses in molecular detection and quantification.


Assuntos
Lacase , Luminol , Luminol/química , Lacase/química , Luminescência , Medições Luminescentes
2.
Drug Chem Toxicol ; 45(6): 2653-2663, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34595971

RESUMO

This study evaluated the in vitro antimicrobial and immunomodulatory action of crude extracts from Anacardium occidentale L. (cashew tree) leaves and bark, and to determine their toxicity to peripheral-blood mononuclear cells (PBMCs) and to zebrafish embryos and larvae. Chemical analysis of extracts was performed by proton nuclear magnetic resonance (1H-NMR). The antibacterial activity was evaluated against selected bacteria strains by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Cytotoxicity of the extracts was assessed using resazurin method, while the effect on production of ROS by PMN leukocytes was measured by luminol. Embryotoxicity to zebrafish was assessed using the fish embryo acute toxicity test (FET) and quantification of toxicity marker enzymes (AChE, LDH, and GST). 1H-NMR results showed anacardic acid as the main component of the extracts. All bacterial species tested were sensitive to the extracts, with MICs ranging from 312.5 to 10,000 µg/mL. Streptococcus mutans and Escherichia coli were the most susceptible species. The extracts promoted cell viability above 75% at concentrations from 1.25 to 80 µg/mL. Both extracts reduced zymosan-induced ROS (p < 0.05) at concentrations of 1, 8, and 80 µg/mL compared to the control. In vivo, there were embryotoxic effects in zebrafish embryos exposed to both extracts through the presence of lethal and sublethal endpoints. The samples also acted by inhibiting the activities of biomarker enzymes. The A. occidentale L. bark and leaf extracts showed antimicrobial potential and modulated ROS production in vitro, but these also showed embryotoxic effects to zebrafish.


Assuntos
Anacardium , Animais , Anacardium/química , Peixe-Zebra , Luminol , Zimosan , Prótons , Espécies Reativas de Oxigênio , Extratos Vegetais/toxicidade , Extratos Vegetais/química , Antibacterianos/toxicidade , Antibacterianos/química , Bactérias , Anti-Inflamatórios , Leucócitos
3.
Photochem Photobiol ; 97(5): 963-970, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33657673

RESUMO

Singlet oxygen (1 O2 ) is the "active principle" in photodynamic therapy. Taurine chloramine (Tau-NHCl) and hydrogen peroxide (H2 O2 ) are well-tolerated and widely used antiseptics. Due to its mild oxidizing features and stability, Tau-NHCl can be directly used to treat skin diseases. We found that a diluted aqueous mixture of Tau-NHCl and H2 O2 acts as a slow and long-lasting potential source of 1 O2 . The reactions were studied by luminol-enhanced chemiluminescence. Evidence of the formation of 1 O2 was obtained using deuterium oxide, sodium azide and 9,10-Anthracenediyl-bis(methylene)dimalonic acid, a chemical trap of 1 O2 . The reaction was optimized, and a mechanism was proposed, including theoretical calculations at B3LYP/6-311++G(3df,2p) level of theory, adding D3Bj empirical dispersion and SMD (Water) solvent effects. Chloramines produced by the reactions between HOCl and L-alanine, 3-amino-1-propanesulfonic acid and gamma-aminobutyric acid were also prepared, and their reactivity and stability were compared with Tau-NHCl. We found that Tau-NHCl is more stable and adequate for the production of 1 O2 . In conclusion, we propose applying these drugs combination as a potential source of 1 O2 with applications for skin diseases treatment.


Assuntos
Peróxido de Hidrogênio , Oxigênio Singlete , Cloraminas , Luminol , Oxigênio , Taurina/análogos & derivados
4.
PLoS Negl Trop Dis ; 14(7): e0008500, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32730339

RESUMO

Schistosomiasis remains a serious public health problem in tropical regions, affecting more than 250 million people. Sensitive diagnostic methods represent key tools for disease elimination, in particular in areas with low endemicity. Advances in the use of luminol-based chemiluminescent techniques have enabled greater sensitivity and speed in obtaining results in different diagnostic settings. In this study, we developed a luminol-H2O2 chemiluminescence (CL) method to detect Schistosoma mansoni eggs in human fecal sediments processed by the Helmintex (HTX) method. After S. mansoni eggs were incubated with a solution of luminol-H2O2 the light emission was detected and measured by spectrophotometry at 431 nm for 5 min, using detection and counts of eggs by bright field optical microscopy as a reference. CL intensity was found to correlate with different sources and numbers of eggs. Furthermore, our results showed that the CL method can distinguish positive from negative samples with 100% sensitivity and 71% specificity. To our knowledge, this is the first study to report the use of CL for the diagnosis of helminths from fecal samples. The combination of the HTX method with CL represents an important advance in providing a reference method with the highest standards of sensitivity.


Assuntos
Fezes/parasitologia , Peróxido de Hidrogênio/química , Luminol/química , Óvulo , Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/diagnóstico , Animais , Humanos , Medições Luminescentes , Camundongos , Esquistossomose mansoni/parasitologia
5.
Photochem Photobiol ; 95(1): 430-438, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30357853

RESUMO

Inflammatory cells, most especially neutrophils, can be a necessary component of the antitumor activity occurring after administration of photodynamic therapy. Generation of neutrophil responses has been suggested to be particularly important in instances when the delivered photodynamic therapy (PDT) dose is insufficient. In these cases, the release of neutrophil granules and engagement of antitumor immunity may play an important role in eliminating residual disease. Herein, we utilize in vivo imaging of luminol chemiluminescence to noninvasively monitor neutrophil activation after PDT administration. Studies were performed in the AB12 murine model of mesothelioma, treated with Photofrin-PDT. Luminol-generated chemiluminescence increased transiently 1 h after PDT, followed by a subsequent decrease at 4 h after PDT. The production of luminol signal was not associated with the influx of Ly6G+ cells, but was related to oxidative burst, as an indicator of neutrophil function. Most importantly, greater levels of luminol chemiluminescence 1 h after PDT were prognostic of a complete response at 90 days after PDT. Taken together, this research supports an important role for early activity by Ly6G+ cells in the generation of long-term PDT responses in mesothelioma, and it points to luminol chemiluminescence as a potentially useful approach for preclinical monitoring of neutrophil activation by PDT.


Assuntos
Luminol/química , Mesotelioma/tratamento farmacológico , Neutrófilos/efeitos dos fármacos , Fotoquimioterapia , Animais , Biomarcadores/metabolismo , Éter de Diematoporfirina/uso terapêutico , Luminescência , Mesotelioma/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ativação de Neutrófilo/efeitos dos fármacos , Neutrófilos/imunologia , Fármacos Fotossensibilizantes/uso terapêutico , Prognóstico
6.
Chemosphere ; 196: 361-367, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29310073

RESUMO

A soft material formed by multiwall carbon nanotubes and 1-butyl-3-methyl imidazolium chloride was used as sorbent material to perform the chromium speciation in natural waters. This soft material was not yet used for the speciation of metals as chromium. Thus, a multicommutated flow system containing a minicolumn packed with the soft material was designed. The procedure was based on the capacity of the sorbent to retain Cr(VI) as Cr2O7= and allow to pass Cr(III) through the column. Then, a fully automated flow-batch analysis system was developed to quantify both species using chemiluminescence detection. Thus, Cr(III) was determined as catalyst of the luminol and hydrogen peroxide reaction and Cr(VI) as oxidant of luminol reaction. This represents a new approach because the oxidation of luminol using Cr2O7= has not been reported in literature. The variables of the two systems were optimized. The limits of detection were 1.4 µg L-1 for Cr(VI) and 4.0 µg L-1 for Cr(III). The precision of the method was 3.8% and 7.0% for Cr(VI) and Cr(III), respectively. The present method was applied to real water samples with recoveries between 95% and 107%. Besides, these results were in accordance with those obtained using inductive coupled plasma-optical emission spectrometry technique.


Assuntos
Cromo/química , Luminescência , Automação , Cromo/análise , Desenho de Equipamento , Peróxido de Hidrogênio/química , Concentração de Íons de Hidrogênio , Luminol , Nanotubos de Carbono , Oxirredução , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/química
7.
Rev. odontol. UNESP (Online) ; 46(4): 227-231, July-Aug. 2017. tab, ilus
Artigo em Inglês | LILACS, BBO - Odontologia | ID: biblio-902656

RESUMO

Introduction: Blood is a biological material with high potential of infectious transmission in dental environments, including herpes simplex, hepatitis and AIDS. Aim: To investigate the efficacy of luminol in detecting blood in endodontic files before and after the sterilization process. Material and method: Luminol was used to investigate the presence or absence of traces of blood tissue in 50 endodontic files, visible to naked eye or not, after performing endodontic treatment and after the cleaning/sterilization process. The results obtained were tabulated and statistically analyzed by using the Friedman's test at a significance level of 5% (p<0.05). Result: By naked eye, it was found that 31/50 files showed no trace of blood, 8/50 showed a slight presence of blood and 11/50 showed a considerable presence of blood after endodontic treatment. After the use of luminol, however, 16/50 endodontic files showed no trace of blood, 19/50 showed a slight presence of blood and 15/50 showed a considerable presence of blood. After the cleaning and sterilization process, no blood was detected in the files. Conclusion: It was concluded that the luminol solution is effective in detecting blood tissue in endodontic files as well as in validating the cleaning/sterilization process.


Introdução: Sangue é um material biológico com alto potencial de transmissão de infecção em ambientes odontológicos, incluindo herpes simples, hepatites e AIDS. Objetivo: Investigar a eficácia do luminol em detector sangue em limas endodônticas antes e após o processo de esterilização. Material e método: Luminol foi utilizado para investigar a presença ou ausência de vestígios tecido sanguíneo em 50 limas endodônticas, visíveis ou não à olho nu, após a realização do tratamento endodôntico e após o processo de limpeza/esterilização. Os resultados obtidos foram tabulados e analisados estatisticamente utilizando o teste de Friedman com nível de significância de 5% (p<0,05). Resultado: A olho nú, foi observado que 31/50 limas não apresentaram vestígios de sangue, 8/50 apresentaram uma leve presença de sangue e 11/50 apresentaram uma presença considerável de sangue após o tratamento endodôntico. Após a utilização do luminol, entretanto, 16/50 limas endodônticas não apresentaram vestígios de sangue, 19/50 apresentaram uma leve presença de sangue e 15/50 apresentaram uma presença considerável de sangue. Após o processo de limpeza e esterilização não foi detectado sangue nas limas endodônticas. Conclusão: A solução de luminol é efetiva na detecção de tecido sanguíneo em limas endodônticas, validando o processo de limpeza/esterilização.


Assuntos
Sangue , Esterilização , Controle de Infecções , Clínicas Odontológicas , Endodontia/instrumentação , Luminol , Terapêutica , Síndrome da Imunodeficiência Adquirida , Hepatite , Herpes Zoster
8.
Talanta ; 135: 27-33, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25640122

RESUMO

A new chemiluminescence (CL) flow method for persulfate determination was developed based on luminol oxidation by in-line generated radicals. Reactive oxygen species (ROS) generated by CdTe quantum dots (QDs) under a low energetic radiation (visible light emitted by LEDs) promoted the decomposition of persulfate ion (S2O8(2-)) into sulfate radical (SO4(∙-)), leading to subsequent radical chain reactions that yield the emission of light. Due to the inherent radical short lifetimes and the transient behavior of CL phenomena an automated multi-pumping flow system (MPFS) was proposed to improve sample manipulation and reaction zone implementation ensuring reproducible analysis time and high sampling rate. The developed approach allowed up to 60 determinations per hour and determine S2O8(2-) concentrations between 0.1 and 1 mmol with good linearity (R=0.9999). The method has shown good repeatability with relative standard deviations below 2.5% (n=3) for different persulfate concentrations (0.1 and 0.625 mmol L(-1)). Limits of detection (3σ) and quantification (10σ) were 2.7 and 9.1 µmol L(-1), respectively. The MPFS system was applied to persulfate determination in bench scale UV/S2O8(2-) drug degradation processes of model samples showing good versatility and providing real time information on the persulfate consumption in photo-chemical degradation methodologies.


Assuntos
Ácido 3-Mercaptopropiônico/efeitos da radiação , Compostos de Cádmio/efeitos da radiação , Luminol/química , Compostos de Potássio/efeitos da radiação , Pontos Quânticos/efeitos da radiação , Espécies Reativas de Oxigênio/química , Sulfatos/efeitos da radiação , Telúrio/efeitos da radiação , Ácido 3-Mercaptopropiônico/química , Compostos de Cádmio/química , Luz , Nanopartículas/química , Oxirredução , Compostos de Potássio/química , Pontos Quânticos/química , Sulfatos/química , Telúrio/química
9.
Luminescence ; 29(7): 901-7, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24585556

RESUMO

An automated multi-pumping flow system is proposed for the chemiluminometric determination of ascorbic acid in pharmaceutical formulations, relying on the ability of semiconductor nanocrystals to generate short-lived reactive species upon photo-irradiation. A photo-unit based on visible-light-emitting diodes is used to photo-excite cadmium telluride (CdTe) quantum dots capped with glutathione, leading to the generation of radicals that react with luminol under alkaline conditions, yielding the chemiluminescence. Ascorbic acid acts as a radical scavenger, preventing the oxidation of luminol, thus ensuring a concentration-dependent chemiluminescence quenching. After system optimization, a linear working range of 5.0 × 10(-7) to 5.0 × 10(-6) mol/L ascorbic acid (r = 0.9967, n = 5) was attained, with a detection limit of 3.05 × 10(-7) mol/L and a sampling rate of 200/h. The flow system was applied to the analysis of pharmaceutical formulations and the results were in good agreement with those obtained by the reference titrimetric procedure (RD < ± 4.3%, n = 7).


Assuntos
Ácido Ascórbico/análise , Compostos de Cádmio/química , Glutationa/química , Pontos Quânticos , Telúrio/química , Química Farmacêutica , Medições Luminescentes , Luminol/química , Estrutura Molecular , Processos Fotoquímicos , Espécies Reativas de Oxigênio
10.
J Microencapsul ; 28(4): 258-67, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21545317

RESUMO

Considering that antioxidant flavonols have been reported to be beneficial to human health, but that their low water solubility and bioavailability limit their administration through systemic route, the development of suitable flavonol-carriers is of great importance for clinical therapeutics. The aim of this study was to prepare liposomes containing flavonols or not and evaluate their antioxidant activity. Vesicles were obtained by ethanol injection method and characterized in terms of entrapment efficiency, size and zeta potential. Inhibitory activity of liposomal flavonols on reactive oxygen species generation was assessed in vitro using luminol-H(2)O(2)-horseradish peroxidase technique. Antioxidant activity of liposomal flavonols is dependent on concentration and chemical structure of active compound. Quercetin and myricetin are the most active flavonols (IC(50) = 0.6-0.9 µmol/L), followed by kaempferol (IC(50) = 3.0-4.5 µmol/L) and galangin (IC(50) = 4.0-7.0 µmol/L). Our results suggest that antioxidant-loaded liposomes may be promising tools for therapy of diseases where oxidative stress is involved.


Assuntos
Antioxidantes/química , Flavonóis/química , Peróxido de Hidrogênio/química , Luminol/química , Avaliação Pré-Clínica de Medicamentos , Peroxidase do Rábano Silvestre/química , Humanos , Lipossomos
11.
Forensic Sci Int ; 206(1-3): 58-61, 2011 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-20643520

RESUMO

Bloodstains often constitute the major physical evidence in crime investigation. Diluted blood invisible to the naked eye can be detected through presumptive tests however such tests can damage samples and prevent further processing such as DNA analysis. In this study, we compared the effects of luminol (prepared according to Weber [15]), Luminol 16(®), Bluestar(®) and benzidine for inhibition in the human antiglobulin test and the human hemoglobin immunochromatographic test and on the total human DNA concentration up to 120 days after sample treatment. Treatment with both luminol solutions and Bluestar(®) still allowed positive results for the immunologic tests, indicating non-interference with human blood confirmatory tests. However, samples treated with benzidine could not be further analyzed by serological tests. Also, DNA quantification showed that 48h after benzidine treatment, but not luminol or Bluestar solution application, sample DNA was degraded. Luminol 16(®) caused DNA degradation already at 30 days post-application. At 120 days post treatment, all samples treated with any of the agents but not untreated samples had DNA degradation.


Assuntos
Degradação Necrótica do DNA/efeitos dos fármacos , DNA/análise , Reação em Cadeia da Polimerase/métodos , Testes Sorológicos , Benzidinas/química , Manchas de Sangue , Humanos , Indicadores e Reagentes/química , Luminol/química , Manejo de Espécimes , Fatores de Tempo
12.
Biocell ; Biocell;34(3): 121-132, Dec. 2010. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-595013

RESUMO

Wilms tumor gene 1 (wt-1), a key regulator of mesenchymal-epithelial transformation, is downregulated during congenital obstructive nephropathy, leading to apoptosis. There is a functional interaction between WT-1 and inducible nitric oxide synthase (iNOS). In this regard, we reported that after neonatal unilateral ureteral obstruction, rosuvastatin prevents apoptosis through an increase in nitric oxide bioavailability, which in turn is linked to higher Hsp70 expression. Hence, the goal of this study was to determine whether a nitric oxide/Hsp70 interaction is involved in changes in WT-1 mRNA expression after ureteral obstruction. Neonatal rats submitted to experimental ureteral obstruction were treated with either vehicle or rosuvastatin for 14 days. Decreased nitric oxide and iNOS/Hsp70 expression associated wit h WT-1 low expression was shown in obstructed kidneys. Apoptosis was induced and it was associated with an increased Bax/BcL2 ratio. Conversely, iNOS/Hsp70 upregulation and an increased WT-1 mRNA expression, without an apoptotic response, were observed in the cortex of obstructed kidneys of rosuvastatin-treated rats. Nitric oxide also modulated Hsp70 and WT-1 mRNA expression in MDCK cells. Finally, in vivo experiments with nitric oxide modulators support our hypothesis that WT-1 mRNA expression is associated with nitric oxide level. Results suggest that rosuvastatin may modulate WT-1 mRNA expression through renal nitric oxide bioavailability, preventing neonatal obstruction-induced apoptosis associated with Hsp70 interaction.


Assuntos
Masculino , Animais , Feminino , Recém-Nascido , Cães , Ratos , Apoptose , Apoptose/fisiologia , Linhagem Celular , Células Epiteliais/citologia , Células Epiteliais , Células Epiteliais/fisiologia , Modelos Animais de Doenças , Inibidores Enzimáticos/farmacologia , Luminol/análogos & derivados , Luminol/farmacologia , Fluorbenzenos/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , /genética , /metabolismo , Rim/citologia
13.
Biocell ; Biocell;34(3): 121-132, Dec. 2010. ilus, graf, tab
Artigo em Inglês | BINACIS | ID: bin-127242

RESUMO

Wilms tumor gene 1 (wt-1), a key regulator of mesenchymal-epithelial transformation, is downregulated during congenital obstructive nephropathy, leading to apoptosis. There is a functional interaction between WT-1 and inducible nitric oxide synthase (iNOS). In this regard, we reported that after neonatal unilateral ureteral obstruction, rosuvastatin prevents apoptosis through an increase in nitric oxide bioavailability, which in turn is linked to higher Hsp70 expression. Hence, the goal of this study was to determine whether a nitric oxide/Hsp70 interaction is involved in changes in WT-1 mRNA expression after ureteral obstruction. Neonatal rats submitted to experimental ureteral obstruction were treated with either vehicle or rosuvastatin for 14 days. Decreased nitric oxide and iNOS/Hsp70 expression associated wit h WT-1 low expression was shown in obstructed kidneys. Apoptosis was induced and it was associated with an increased Bax/BcL2 ratio. Conversely, iNOS/Hsp70 upregulation and an increased WT-1 mRNA expression, without an apoptotic response, were observed in the cortex of obstructed kidneys of rosuvastatin-treated rats. Nitric oxide also modulated Hsp70 and WT-1 mRNA expression in MDCK cells. Finally, in vivo experiments with nitric oxide modulators support our hypothesis that WT-1 mRNA expression is associated with nitric oxide level. Results suggest that rosuvastatin may modulate WT-1 mRNA expression through renal nitric oxide bioavailability, preventing neonatal obstruction-induced apoptosis associated with Hsp70 interaction.(AU)


Assuntos
Masculino , Animais , Feminino , Recém-Nascido , Cães , Ratos , Apoptose , Apoptose/fisiologia , Linhagem Celular , Modelos Animais de Doenças , Inibidores Enzimáticos/farmacologia , Células Epiteliais/citologia , Células Epiteliais , Células Epiteliais/fisiologia , Luminol/análogos & derivados , Luminol/farmacologia , Fluorbenzenos/farmacologia , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Rim/citologia
14.
Biocell ; 34(3): 121-32, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21443142

RESUMO

Wilms tumor gene 1 (wt-1), a key regulator of mesenchymal-epithelial transformation, is downregulated during congenital obstructive nephropathy, leading to apoptosis. There is a functional interaction between WT-1 and inducible nitric oxide synthase (iNOS). In this regard, we reported that after neonatal unilateral ureteral obstruction, rosuvastatin prevents apoptosis through an increase in nitric oxide bioavailability, which in turn is linked to higher Hsp70 expression. Hence, the goal of this study was to determine whether a nitric oxide/Hsp70 interaction is involved in changes in WT-1 mRNA expression after ureteral obstruction. Neonatal rats submitted to experimental ureteral obstruction were treated with either vehicle or rosuvastatin for 14 days. Decreased nitric oxide and iNOS/Hsp70 expression associated with WT-1 low expression was shown in obstructed kidneys. Apoptosis was induced and it was associated with an increased Bax/BcL2 ratio. Conversely, iNOS/Hsp70 upregulation and an increased WT-1 mRNA expression, without an apoptotic response, were observed in the cortex of obstructed kidneys of rosuvastatin-treated rats. Nitric oxide also modulated Hsp70 and WT-1 mRNA expression in MDCK cells. Finally, in vivo experiments with nitric oxide modulators support our hypothesis that WT-1 mRNA expression is associated with nitric oxide level. Results suggest that rosuvastatin may modulate WT-1 mRNA expression through renal nitric oxide bioavailability, preventing neonatal obstruction-induced apoptosis associated with Hsp70 interaction.


Assuntos
Proteínas de Choque Térmico HSP70/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico/metabolismo , Obstrução Ureteral/fisiopatologia , Proteínas WT1/genética , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Linhagem Celular , Modelos Animais de Doenças , Cães , Inibidores Enzimáticos/farmacologia , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Feminino , Fluorbenzenos/farmacologia , Proteínas de Choque Térmico HSP70/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Rim/citologia , Luminol/análogos & derivados , Luminol/farmacologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Oligopeptídeos/farmacologia , Pirimidinas/farmacologia , Ratos , Ratos Endogâmicos WKY , Rosuvastatina Cálcica , Sulfonamidas/farmacologia , Proteína Supressora de Tumor p53/genética , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia , Proteínas WT1/metabolismo
15.
Anal Biochem ; 385(1): 107-14, 2009 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-19027710

RESUMO

The total reactive antioxidant potential (TRAP) is one of the methods most employed to estimate the antioxidant capacity of samples in vitro. This method is based on the quenching of luminol-enhanced chemiluminescence derived from the thermolysis of 2,2'-azo-bis(2-amidinopropane)dihydrochloride (AAPH) as the free radical source. However, this method can present limitations when the sample does not present a lag phase. In addition, there are no studies regarding TRAP assay validation. In this context, the aim of this work was to optimize and validate this method and to propose another evaluation method using the area under the curve (AUC). The main condition established was the need for the stabilization of the system, at 7000s, before the addition of the antioxidant to be tested. Both evaluation methods were validated using Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) as a calibrator in the range of 50 to 250nM, and all parameters showed satisfactory results: specificity, linearity (r>0.99), precision (intra and interassay relative standard deviations <5%), robustness, and the limits of detection and quantitation (low and similar for both methods). The main advantage of the use of AUC is to evaluate the antioxidant potential of samples that do not present lag phase.


Assuntos
Antioxidantes/análise , Técnicas de Química Analítica/métodos , Medições Luminescentes/métodos , Amidinas/química , Área Sob a Curva , Calibragem , Cromanos/química , Radicais Livres/química , Modelos Lineares , Luminol/química , Sensibilidade e Especificidade , Termodinâmica , Fatores de Tempo
16.
Luminescence ; 23(6): 410-6, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18814188

RESUMO

Liquid-core waveguides (LCWs), devices that constrain the emitted radiation minimizing losses during the transport, are an alternative to maximize the amount of detected radiation in luminescence. In this work, the performance of a LCW flow-cell was critically evaluated for chemiluminescence measurements, by using as model the oxidation of luminol by hydrogen peroxide or hypochlorite. An analytical procedure for hypochlorite determination was also developed, with linear response in the range 0.2-3.8 mg/L (2.7-51 micromol/L), a detection limit estimated as 8 microg/L (0.64 micromol/L) at the 99.7% confidence level and luminol consumption of 50 microg/determination. The coefficients of variation were 3.3% and 1.6% for 0.4 and 1.9 mg/L ClO(-), respectively, with a sampling rate of 164 determinations/h. The procedure was applied to the analysis of Dakin's solution samples, yielding results in agreement with those obtained by iodometric titration at the 95% confidence level.


Assuntos
Análise de Injeção de Fluxo/métodos , Peróxido de Hidrogênio/análise , Ácido Hipocloroso/análise , Medições Luminescentes/métodos , Luminol/química , Ferricianetos/química , Análise de Injeção de Fluxo/instrumentação , Luminescência , Medições Luminescentes/instrumentação , Oxirredução , Sensibilidade e Especificidade , Fatores de Tempo , Titulometria/métodos
17.
Pharmazie ; 63(1): 67-70, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18271307

RESUMO

The tissue damage found in some inflammatory and autoimmune diseases has been shown to be mediated by an increased activation of neutrophil effector functions. In this study, we investigated the inhibitory effect of the phenolic compound butylated hydroxytoluene (BHT) on reactive oxygen species (ROS) generation by opsonized zymosan-stimulated neutrophils, assessed by luminol- and lucigenin-enhanced chemiluminescence (CL-lum and CL-luc, respectively), and some aspects of its mechanism of action. BHT showed concentration-dependent: (a) inhibitory effect on CL-lum and CL-luc; (b) cytotoxic effect, expressed by increased lactate dehydrogenase leakage by the cells; (c) interaction with neutrophil membranes; (d) ROS scavenger activity. These biological effects were observed in the same range of concentrations (0-5 x 10(-5) mol/l). Taken together, the results suggest that inhibition of neutrophil chemiluminescence by BHT was a result of multiple mechanisms, especially a cytotoxic effect probably mediated by BHT interaction with neutrophils membranes, and the ROS scavenging effect.


Assuntos
Antioxidantes/farmacologia , Hidroxitolueno Butilado/farmacologia , Neutrófilos/efeitos dos fármacos , Acridinas/química , Análise de Variância , Animais , Membrana Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Técnicas In Vitro , L-Lactato Desidrogenase/metabolismo , Luminescência , Luminol/química , Fluidez de Membrana/efeitos dos fármacos , Neutrófilos/enzimologia , Coelhos , Espécies Reativas de Oxigênio/metabolismo
18.
Life Sci ; 81(4): 317-26, 2007 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-17610907

RESUMO

Tissue damage in autoimmune diseases involves excessive production of reactive oxygen species (ROS) triggered by immune complexes (IC) and neutrophil (PMN) interactions via receptors for the Fc portion of IgG (FcgammaR) and complement receptors (CR). Modulation of both the effector potential of these receptors and ROS generation may be relevant to the maintenance of body homeostasis. In the present study, the modulatory effect of four flavonols (myricetin, quercetin, kaempferol, galangin) on rabbit PMN oxidative metabolism, specifically stimulated via FcgammaR, CR or both classes of receptors, was evaluated by luminol- and lucigenin-dependent chemiluminescence assays. Results showed that flavonol inhibitory effect was not dependent on the cell membrane receptor class stimulated but related to the lipophilicity of the compounds (their apparent partition coefficient values were obtained by high-performance liquid chromatography), and was also inversely related to the number of hydroxyl groups in the flavonol B ring and the ROS-scavenger activity (assessed by the luminol--H2O2--horseradish peroxidase reaction). Under the experimental conditions the flavonols tested were not toxic to PMNs (evaluated by lactate dehydrogenase release and trypan blue exclusion) and did not interfere with IC-induced phagocytosis (evaluated by transmission electron microscopy). Our results suggested that inhibition of IC-stimulated PMNs effector functions by the flavonols tested herein was the result of cooperation of different cellular mechanisms.


Assuntos
Derivados de Benzeno/farmacologia , Flavonóis/farmacologia , Fatores Imunológicos/química , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Receptores de Complemento/metabolismo , Receptores Fc/metabolismo , Acridinas/química , Animais , Complexo Antígeno-Anticorpo/imunologia , Complexo Antígeno-Anticorpo/metabolismo , Derivados de Benzeno/química , Derivados de Benzeno/metabolismo , Proteínas do Sistema Complemento/metabolismo , Flavonoides/química , Flavonoides/metabolismo , Flavonoides/farmacologia , Flavonóis/química , Flavonóis/metabolismo , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/metabolismo , Sequestradores de Radicais Livres/farmacologia , Interações Hidrofóbicas e Hidrofílicas , Hidroxilação , Doenças do Complexo Imune/tratamento farmacológico , Doenças do Complexo Imune/metabolismo , Fatores Imunológicos/metabolismo , Quempferóis/química , Quempferóis/metabolismo , Quempferóis/farmacologia , Medições Luminescentes , Luminol/química , Estrutura Molecular , Neutrófilos/imunologia , Oxirredução/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Fagocitose/imunologia , Quercetina/química , Quercetina/metabolismo , Quercetina/farmacologia , Coelhos , Receptores de Complemento/imunologia , Receptores Fc/imunologia , Relação Estrutura-Atividade
19.
Luminescence ; 22(2): 126-33, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17089344

RESUMO

The use of poly(N-vinyl-2-pyrrolidone) (PVP) hydrogel-supported luminol chemiluminescence (CL) for the automatic determination of hydrogen peroxide and the quantification of the antiradical capacity of Trolox is described. The hydrogel containing luminol and hemin is prepared directly on a 96-well microplate and can be stored for up to 3 months without significant decrease in CL quantum yields. Furthermore, this system can also be used as a secondary light standard for the calibration of microplate luminometers.


Assuntos
Hidrogéis/química , Peróxido de Hidrogênio/análise , Medições Luminescentes/instrumentação , Luminol/química , Povidona/química , Antioxidantes/química , Antioxidantes/farmacologia , Automação , Calibragem , Cromanos/química , Cromanos/farmacologia , Hemina/química , Medições Luminescentes/métodos , Estrutura Molecular
20.
Luminescence ; 22(2): 113-25, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17089370

RESUMO

The chemiluminescent oxidation of luminol by hydrogen peroxide in the presence of hemin is revisited in an UV-C cross-linked PVP hydrogel. Chemiluminescence properties such as initial light intensity (I(0)), area of emission (S) and observed rate constants (k(obs)) are studied, varying the concentration of all reactants using a multivariate factorial approach.


Assuntos
Hemina/química , Hidrogéis/síntese química , Luminescência , Luminol/química , Povidona/química , Reagentes de Ligações Cruzadas/química , Análise Fatorial , Hemina/efeitos da radiação , Hidrogéis/química , Peróxido de Hidrogênio/química , Concentração de Íons de Hidrogênio , Cinética , Luminol/efeitos da radiação , Estrutura Molecular , Oxirredução , Povidona/efeitos da radiação , Raios Ultravioleta
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