DCFH-DA

Top PDF DCFH-DA:

Journal of Applied Pharmaceutical Science

Journal of Applied Pharmaceutical Science

Iso-caryophyllene, a sesquiterpene, is present in several essential oils from vegetable species. In previous work, iso-caryophyllene was found cytotoxic against in vitro culture cell lines but its mechanism of action is still unknown. Reactive oxygen species (ROS) and lipid oxidation induced by iso-caryophyllene were assessed using DCFH-DA and BODIPY-C11, respectively. The results show that iso-caryophyllene induces significant overproduction of ROS by about 187 % at 100 µM and lipid oxidation, which are both partially inhibited by - tocopherol. The effect of isocaryophyllene on membrane permeabilization was evaluated using calcein-AM assay that show that iso-caryophyllene causes membrane permeabilization and cell shrinking. -Tocopherol significantly prevents membrane permeabilization, cell shrinking and cell death, suggesting that lipid oxidation is in part implied in the cytotoxicity. Electrochemical experiments indicate that the superoxide anion reacts with iso-caryophyllene and possibly form oxidized derivatives which could initiate lipid oxidation. Interestingly, superoxide anion reacts more readily with -tocopherol in comparison with iso-caryophyllene which could explain its protective effect on cellular membrane.
Show more

7 Read more

Protective effects of γ-aminobutyric acid against H2O2-induced oxidative stress in RIN-m5F pancreatic cells

Protective effects of γ-aminobutyric acid against H2O2-induced oxidative stress in RIN-m5F pancreatic cells

Viable cells were harvested, and dichloro-dihydro-fluorescein diacetate (DCFH-DA) was used to detect reactive oxygen species (ROS) level; cellular redox status and insulin secretion were[r]

9 Read more

Meripilus giganteus ethanolic extract exhibits pro apoptotic and anti proliferative effects in leukemic cell lines

Meripilus giganteus ethanolic extract exhibits pro apoptotic and anti proliferative effects in leukemic cell lines

After 24 h treatment Jurkat and HL-60 cells (1.00 × 10 6 cells/mL), were washed twice in HBSS, and incubated with 5 μM 2′-7′-dichlorodihydrofluorescin diacetate (DCFH-DA) (Sigma-Aldrich, Saint Louis, MO, USA), for 20 min at 37 °C. When inside the cell, DCFH-DA is dea- cetylated and can be oxidized by ROS to the highly fluorescent 2′,7′-dichlorofluorescein (DCF). DCF fluor- escence was measured using a multi-well plate reader (Wallac Victor2, PerkinElmer) at excitation and emission wavelengths of 485 and 535 nm, respectively [27]. Fluor- escence values were reported as percentage of intracellu- lar ROS with respect to control.
Show more

14 Read more

Metal-based NanoEnhancers for Future Radiotherapy: Radiosensitizing and Synergistic Effects on Tumor Cells

Metal-based NanoEnhancers for Future Radiotherapy: Radiosensitizing and Synergistic Effects on Tumor Cells

3-methyladenine; 53BP1: p53-binding protein 1; AFM: atomic force microscopy; AgNPs: silver nanoparticles; AGuIX: Activation and Guidance of Irradiation by X-ray; AuNCs: gold nanoclusters; AuNPs: gold nanoparticles; Bax: Bcl-2-associated X protein; Bcl-2: B-cell lymphoma-2; BiNPs: bismuth nanoparticles; BRCA1: breast cancer 1; BSA: bovine serum albumin; C18PMH: poly(maleic anhydride-alt-1-octadecene); c-Abl: cellular Abelson; CAT: catalase; CDK: cyclin-dependent kinases; CF: conventional fractionation; CFP: cyan fluorescent protein; CLIONs: cross-linked dextran-coated IONs; CQ: chloroquine; CR: Čerenkov radiation; CT: computed tomography; CTCs: circulating tumor cells; DCFH-DA: 2’,7’-dichlorofluorescein diacetate; DDR: DNA damage-response; DDS: drug delivery systems; DEF: radiation dose enhancement factor; DMPK: drug metabolism and pharmacokinetic; DMSO: dimethyl sulfoxide; DOTA: 1,4,7,10-tetraazacyclododecane- N,N',N'',N'''-tetraacetic acid; DOTAGA: 1,4,7,10- tetraazacyclododecane-1-glutaric anhydride-4,7,10- triacetic acid; DQ-BSA: derivative-quenched bovine serum albumin; DSBs: double-strand breaks; DTDTPA: dithiolated derivative of the diethylenetriaminepentacetic acid; DTPA: diethylenetriaminepentaacetic acid; DTX: docetaxel; EPR: enhanced permeability and retention; ER: endoplasmic reticulum; ESCC: esophageal squamous cell carcinoma; FTIR: Fourier transform infrared; Gd-DTPA: gadopentetic acid; Gd-NCT: gadolinium neutron-capture therapy; GdNPs: gadolinium-based nanoparticles; Gd-tex 2+ : Gd (III) texaphyrin; GeNPs:
Show more

26 Read more

IN-VITRO NEPHROPROTECTIVE ROLE OF ETHANOLIC ROOT EXTRACT OF Boerhaavia diffusa AGAINST CISPLATIN-INDUCED NEPHROTOXICITY

IN-VITRO NEPHROPROTECTIVE ROLE OF ETHANOLIC ROOT EXTRACT OF Boerhaavia diffusa AGAINST CISPLATIN-INDUCED NEPHROTOXICITY

Graph 6: Effect of ERE on ROS induced by cisplatin. LLC-Pk1 was incubated with cisplatin with or without ERE for 24 hours. To measure the intracellular peroxide formation DCFH-DA was used as a probe. This compound reacts with the ROS produced in the cells, oxidized and forms the fluorescent product DCF, this fluorescence product was measured as mentioned in the materials & methods. Experiments were performed in triplicates and repeated twice.

8 Read more

Verbesina encelioides: cytotoxicity, cell cycle arrest, and oxidative DNA damage in human liver cancer (HepG2) cell line

Verbesina encelioides: cytotoxicity, cell cycle arrest, and oxidative DNA damage in human liver cancer (HepG2) cell line

Abbreviations %: percentage; °C: degree celsius; A-549: human lung cancer cell line; CO2: carbon dioxide; DCFH-DA: dichloro-dihydro-fluorescein diacetate; DMEM: Dulbecco’s modified eagle[r]

10 Read more

Conditioned media from adipose stromal cells limit lipopolysaccharide induced lung injury, endothelial hyperpermeability and apoptosis

Conditioned media from adipose stromal cells limit lipopolysaccharide induced lung injury, endothelial hyperpermeability and apoptosis

BALF from saline lungs, neutrophils were the dominant cell type in BALF from LPS-challenged lungs. Although ASC or ASC-CM treatment did not shift the overall neutro- phil/macrophage balance (Figure 5A,B), a significant sup- pression of total WBC count was evident in LPS mice receiving ASC injection (Figure 5A). A similar trend was observed in mice receiving ASC-CM, but did not reach significance at 48 h post-injection (Figure 5B). To further characterize possible mechanisms of ASC-CM-mediated lung injury limitation, we compared the ability of BALF WBC, obtained from LPS- or LPS/ASC-CM-receiving mice, to generate ROS. The method for the measurement of neutrophils oxidative activity was based on ROS- dependent oxidation of DCFH-DA (non-fluorescent) to highly fluorescent DCF. Isolated BALF WBC were tested for their abilities to oxidize DCFH-DA in response to LPS
Show more

15 Read more

Jingfukang induces anti cancer activity through oxidative stress mediated DNA damage in circulating human lung cancer cells

Jingfukang induces anti cancer activity through oxidative stress mediated DNA damage in circulating human lung cancer cells

Jinfukang oral liquid freeze-dried powder was prepared and detected the fingerprint (Additional file 2: Table S2) by Pro- fessor Yu Jin of East China University of Science and Tech- nology (Shanghai, China). A cell counting kit-8 (CCK-8) was obtained from Dojindo. The Annexin V-FITC apoptosis de- tection kit and propidium iodide (PI) were purchased from BD Pharmingen. 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA) and the antioxidant NAC were purchased from Sigma. A caspase-3 detection kit was purchased from Biovi- sion. The anti-γ-H2AX, anti-p-ATM, anti-p-ATR, anti- PARP1, goat anti-mouse IgG-HRP and donkey anti-rabbit IgG-HRP antibodies were purchased from Cell Signaling Technology. The anti-p53, anti-p21, anti-CDK4, anti-Cyclin D, anti-Cyclin E, anti-Fas, anti-Survivin, and anti-β-actin antibodies were purchased from Proteintech.
Show more

8 Read more

Red ginseng abrogates oxidative stress via mitochondria protection mediated by LKB1 AMPK pathway

Red ginseng abrogates oxidative stress via mitochondria protection mediated by LKB1 AMPK pathway

RGE was provided by Korea Tobacco & Ginseng Corporation (Daejeon, Korea) [22]. AA and compound C were purchased from Calbiochem (San Diego, CA). Anti-procaspase-3, anti- phospho-acetyl-CoA carboxylase (ACC), anti-PARP, anti- phospho-LKB1 and anti-phospho-AMPK antibodies were obtained from Cell Signaling Technology (Beverly, MA). Anti-AMPK, anti-ACC and anti-LKB1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase-conjugated goat anti-rabbit, rabbit anti-goat, and goat anti-mouse IgGs were obtained from Zymed Laboratories (San Francisco, CA). Ferric ni- trate, nitrilotriacetic acid [9], 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide (MTT), rhodamine 123, 2 0 ,7 0 -Dichlorofluorescein diacetate (DCFH-DA), anti-β-actin antibody, and other reagents were purchased from Sigma (St. Louis, MO). The solution of iron-NTA complex was prepared as described previously [7].
Show more

9 Read more

<p>In vitro mitochondrial-targeted antioxidant peptide induces apoptosis in cancer cells</p>

<p>In vitro mitochondrial-targeted antioxidant peptide induces apoptosis in cancer cells</p>

Figure 3 The levels of intracellular and mitochondrial ROS in different cell lines after treatment with KRA and the KRSH peptide for 24 h were detected using DCFH-DA and MitoSOX TM.. , a[r]

10 Read more

Intracellular free radical production by peripheral blood T lymphocytes from patients with systemic sclerosis: role of NADPH oxidase and ERK1/2

Intracellular free radical production by peripheral blood T lymphocytes from patients with systemic sclerosis: role of NADPH oxidase and ERK1/2

P <0.05 compared to normal PBL. (B) ROS production by PBL from one healthy control (black line) and one SSc patient (grey line) was analyzed by FACS analysis. A representative histogram of three independent experiments is shown. (C and D) SSc CD14- (C) and SSc CD19- cells (D) were purified from PBL using CD14 and CD19 microbeads. The total fractions of cells (PBL, white bars) and the collected depleted fractions (black bars) were stained with 20 μ M DCFH-DA for 20 minutes, and fluorescence was measured on a plate reader fluorimeter. Data are means ± SD of three independent experiments with cells from three distinct subjects. DCFH-DA, 2 ′ , 7 ′ -dichlorodihydrofluorescin diacetate; PBL, peripheral blood lymphocytes; ROS, reactive oxygen species; SSc, systemic sclerosis.
Show more

12 Read more

Original Article Emodin induces apoptosis of human osteosarcoma cells via mitochondria- and endoplasmic reticulum stress-related pathways

Original Article Emodin induces apoptosis of human osteosarcoma cells via mitochondria- and endoplasmic reticulum stress-related pathways

Abstract: Aim: Emodin showed anti-cancer activity against multiple human malignant tumors by inducing apoptosis. However, the apoptotic inducing effect against human osteosarcoma and related mechanism are still not studied. This study was aimed to investigate them. Methods: Emodin was used to incubate human OS cell U2OS cells at seri- ally diluted concentrations. Hoechst staining was used to evaluate apoptosis; flow cytometry was applied to assess the collapse of mitochondrial membrane potential (MMP); intracellular ROS generation was detected by DCFH-DA staining; endoplasmic reticulum stress activation was examined by western blotting. Results: Cell apoptosis of U2OS cells was induced by emodin incubation in a concentration-dependent manner; MMP collapse and ROS generation were identified at starting concentration of 80 μmol/L of emodin in a concentration-dependent manner. ER stress activation was found at beginning concentration of 40 μmol/L of emodin. The MMP collapse was inhibited while the ER stress was not inhibited by NAC administration. Conclusions: Emodin induces death of human osteosarcoma cells by initiating ROS-dependent mitochondria-induced and ROS-independent ER stress-induced apoptosis.
Show more

8 Read more

“Neuroprotective Effects of Rutin against Hydrogen Peroxide Induced Oxidative Stress in Cerebral Glioma Cell Line” by Gaurav Kumar, Jayanand, Quaisar Perween, Rihan Ansari, Durg Vijay Rai, India.

“Neuroprotective Effects of Rutin against Hydrogen Peroxide Induced Oxidative Stress in Cerebral Glioma Cell Line” by Gaurav Kumar, Jayanand, Quaisar Perween, Rihan Ansari, Durg Vijay Rai, India.

The cellular ROS (reactive oxygen species) accumulation was measured with the help of fluorescent probe DCFH- DA (2, 7- dichloro dihydrofluorescein diacetate). 17 DCFH- DA is a nonfluorescent compound, and it can be enzymatically converted to highly fluorescent compound, DCF, in the presence of ROS. After exposure to 500 µM hydrogen peroxide, with or without rutin, the cells (10 6 /ml) were incubated in 2ml of 140 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 5.6 mM glucose, 1.5 mM CaCl 2 , and 20

5 Read more

Reforma da Administração Pública: Antes e Depois da Democracia

Reforma da Administração Pública: Antes e Depois da Democracia

É assim que, além desses possíveis sinónimos, por sua vez, a “reforma” presta-se a múltiplas entradas. Assim, são várias as dimensões pelas quais ou contra às quais se aprecia as reformas. Entre reformas de constitucionalidade de direitos, ou não, ou da organização político-representativa do Estado; reformas vindas ou provocadas do exterior ou por acção própria; com um efusivo entusiasmo inicial ou não; improvisada ou não; dentro ou fora do prazo que se tenha fixada; respostas conjunturais a “crises” ou problemas que requerem uma solução, ou não; essenciais ou descortinadas como acabando por não serem essenciais ou relacionadas ao cumprimento do que o que tenha sido proferido como causa que a tenha justificado; com ou sem a fobia ou a impossibilidade de que todas as alterações não tenham sido contempladas; mais racional ou emotivamente; dependentes somente de um ou formuladas em conjunto; reformas internas (não em conceito de finanças, mas o conceito de planeamento e organização absorve-as, e antecedida de um “re”ou não, a forma, ou o Estado na sua profundidade, compreensão: PDCA) e externas (necessidades da sociedade; SBU do Estado ou o Estado na sua extensão, mais que os deveres do Estado); orgânicas (sistémicas) e mecânicas (organização fechada; a tarefa, o procedimento); maiores ou menores; por acrescentos ou reduções; revolucionárias ou faseadas; sem ou com experiência(s) piloto; globais e parciais, respeitando as parciais a algum processo/produto explorado pela AdP, sector funcional, territorial ou nível da organização política do sistema vigente de organização da sociedade.
Show more

17 Read more

Transtorno do espectro da esquizofrenia: um olhar da família

Transtorno do espectro da esquizofrenia: um olhar da família

2012; Raposo & Pedroso, 2014; Souza Filho, Sousa, Parente, & Martins, 2010; Sanches, 2012; Noto & Nunes, 2012). Assim sendo, a convivência com a esquizofrenia exige dos familiares a transposição das suas necessidades e desejos para um plano secundário e a reorganização das suas vidas de acordo com as necessidades do paciente (Zaneti & Galera, 2007), tendo em vista que o transcorrer do transtorno é marcado por períodos de controle e diminuição dos sintomas, alternados com crises, recaídas e reinternação hospitalar, configurando um cenário de desafios diários (Lopes, 2013) tecidos por dúvidas em relação à situações das mais práticas, até as mais complexas e angustiantes em torno da expectativa de cura. De maneira geral, o desgaste da família ao cuidar de uma pessoa com esquizofrenia deriva, em parte, pelo fato de que as redes de assistência não conseguem atingir essas famílias (Araújo, Coutinho, Viveiros, Leite, & Corrêa, 2015), levando-as à sobrecargas nas rotinas familiares, bem como dificuldades emocionais, físicas e financeiras (Giacon & Galera, 2013), pois, as relações estabelecidas e as práticas de cuidados desenvolvidas no contexto familiar, sofrem influência direta da maneira como o transtorno é percebido pelos famíliares (Behenck, et al., 2011). Pautando-se nisso, os serviços de saúde mental devem promover a articulação entre múltiplas formas assistenciais, múltiplos saberes, para que dessa forma o paciente junto com a família, sejam assistidos integralmente. A priori, a demanda inicial de cuidado é voltada para a pessoa com sofrimento psíquico, no entanto, para a integralidade do cuidado efetivamente, precisa da inclusão da família não somente incentivando o usuário a se envolver no projeto terapêutico, como também participando diretamente das atividades do serviço, tanto internas como nos projetos de trabalho e ações comunitárias de integração social (Brasil, 2004). Partindo dessa perspectiva, de que as famílias também precisam ser alvo de investigação, orientação e cuidado, objetivou-se conhecer como as famílias de pacientes com esquizofrenia compreendem o transtorno. Sendo de suma importância na elaboração de estratégias de cuidado eficazes.
Show more

6 Read more

Análise espacial da centralidade e da dispersão da riqueza gaúcha de 1970 a 2000: notas preliminares

Análise espacial da centralidade e da dispersão da riqueza gaúcha de 1970 a 2000: notas preliminares

Em relação às irregularidades, pode-se frisar o comportamento errático do deslocamento dos centros médios do PIB. Essa situação, em última instância, proporciona uma representação do comportamento geral dessa variável no território gaúcho. Na primeira década (1970-80), o deslocamento aponta na direção de Porto Alegre e Região Metropolitana; entretanto, na década seguinte (1980- -90), o deslocamento é para o norte do Estado, parecendo apontar para a região de Caxias do Sul (uma hipótese); e, na década final (1990-00), o deslocamento muda, novamente, nesse momento direcionado para leste-nordeste, o que pode indicar, relacionado à hipótese anterior, o fortalecimento do crescimento relativo da região de Caxias do Sul. Interessante, também, é observar os deslocamentos dos centros médios da renda mensal per capita: entre 1970 e 1980, o deslocamento vai em direção ao nordeste do Estado; e, na década seguinte (1980-91), sofre uma inflexão em direção ao sul (mais precisamente, leste- -sudeste); e, na década 1991-00, retoma a tendência da década inicial, na qual o centro médio toma o sentido norte-nordeste, o que pode estar indicando o fortalecimento da renda na região da Serra (Caxias do Sul e seu entorno).
Show more

37 Read more

Comparison of fluorescence-based techniques for the quantification of particle-induced hydroxyl radicals

Comparison of fluorescence-based techniques for the quantification of particle-induced hydroxyl radicals

The fluorometer was calibrated with water as zero fluores- cence and a reaction of H 2 O 2 /HRP/glutathione & phenol (for tempo-9-c)/DMSO (for proxyl fluorescamine) as a fluorescence value of 1000. Compared to the fluorescence of only DCFH, APF, and amplex ultrared in a pH buffered water solution, the addition of H 2 O 2 & HRP results in much greater fluorescence intensity. When the fluorome- ter is calibrated with this large difference in fluorescence intensities, subsequent measurements have a greater accu- racy. Both tempo-9-ac and proxyl fluorescamine do not respond with as great of an increase in fluorescence when incubated with H 2 O 2 and HRP. Therefore, the sensitivity of tempo-9-ac and proxyl fluorescamine are lower than the sensitivities of DCFH, APF, and amplex ultrared. The background fluorescence for proxyl fluorescamine is only slightly less than the fluorescence of proxyl fluorescamine reacted with 1 µM H 2 O 2 and HRP (i.e., 890 compared to 1000). When proxyl fluorescamine is incubated with either Fe(II) (Fig. 2) or pyrite (Fig. 3), the larger fluores- cence intensities measured are calculated as significant concentrations of hydroxyl radicals. This problem may be resolved by using more than 1 µM H 2 O 2 in the calibration of proxyl fluorescamine, but that would defeat our objec- tive of comparing all of the probes uniformly.
Show more

9 Read more

Neonatal Brachial Plexus Paralysis: A Discussion on the Light of Literature

Neonatal Brachial Plexus Paralysis: A Discussion on the Light of Literature

Para determinar a severidade da lesão, é importante distinguir dois tipos de lesão: a lesão pré-ganglionar e a pós-ganglionar. A pré-ganglionar ou por avulsão, localiza-se acima da raiz dorsal. A lesão que compromete de C8 a T1, é considerada grave por não ser passível de correção cirúrgica direta, havendo necessidade de estudo por ressonância magnética ou por mielografia para o planejamento da cirurgia. Já a lesão pós-ganglionar é localizada abaixo da raiz dorsal ganglionar, sendo descritos três tipos por Seddon e Sunderland (apud Abid, 2016): neuropraxia, anoxotmese e neurotmese. Na neuropraxia há um bloqueio temporário da condução por alongamento simples, os axônios mantém a integridade, levando a paralisia que involui espontaneamente em duas a três semanas. A axonotmese é caracterizada por ruptura neuronal parcial com integridade da bainha com preservação do perineuro e epineuro, podendo haver recuperação espontânea; no entanto pode ocorrer comutação. Finalmente, a neurotmese que corresponde a ruptura total do nervo periférico, com degeneração walleriana, que pode evoluir com formação de neuroma. É considerada a lesão neuronal periférica mais severa e, assim como nas avulsões, é incomum haver recuperação espontânea (Abid, 2016; Smith, 2018; Alvites, 2018).
Show more

16 Read more

Da Pena Privativa de Liberdade: Controle da Crimilalidade?

Da Pena Privativa de Liberdade: Controle da Crimilalidade?

Além de escravos criminosos condenados, as prisões, como as da ilha das Cobras e Santa Bárbara, detinham escravos enviados para “correção” por seus donos, devido a uma falta cometida por eles, com fugir ou contrair uma doença incurável. Se seus donos nunca assinassem a libertação, eles eram efetivamente abandonados para o resto de suas vidas [...] Esses escravos caíam numa armadilha, porque não podiam sair da prisão sem o consentimento de seus donos e o governo não podia perdoá-los porque não haviam sido condenados por um crime. Portanto, permaneciam na cadeia.
Show more

15 Read more

Representação do aleitamento materno para acadêmicos da área da saúde

Representação do aleitamento materno para acadêmicos da área da saúde

No decorrer das entrevistas, ficou evidente o quanto os acadêmicos enfantizam as vantagens oferecidas através do aleitamento materno. O ponto mais frisado foi sobre a fonte de nutrição, crescimento e desenvolvimento, além da importância por ser um alimento completo. No que tange, os elementos essenciais contidos no leite materno responsáveis pelo crescimento e desenvolvimento, físico e cognitivo do bebê apresentam-se as vitaminas, minerais, ferro, cálcio, magnésio e zinco, esses garantem ao lactente o fortalecimento do sistema imune, protegendo-o contra infecções respiratórias e gastrointestinais, minimizando os riscos de mortalidade infantil (BRASIL, 2015). O leite humano é de suma importância desde os primeiros anos de vida até a fase adulta, pois promove o bom crescimento e desenvolvimento de todo sistema biológico e psíquico, contribuindo assim nas boas condições para a saúde do bebê(SILVA,2017). Nesse contexto, o aleitamento exclusivo oferece uma gama de benefícios, sem qualquer necessidade de complementação com líquidos, fómulas ou alimentos antes dos seis meses de idade da criança
Show more

5 Read more

Show all 6081 documents...