A Microplate-Based Colorimetric Assay of the Total Peroxyl Radical Trapping Capability of Human Plasma

Lussignoli, S.; Fraccaroli, Marta; Andrioli, Giuseppe; Brocco, Giorgio; Bellavite, Paolo

doi:10.1006/abio.1999.4010

Cited by 72 publications

(64 citation statements)

References 34 publications

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“…Crocin was isolated from saffron as previously described method (Lussignoli et al, 1999). Briefly, 500 mg saffron was washed thrice with 20 ml ethyl ether, and the residual ether was evaporated in air.…”

Section: Isolation and Characterization Of Crocin From C Sativusmentioning

confidence: 99%

Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation

Bakshi¹,

Hakkim²,

Sam³

2016

Asian Pacific Journal of Cancer Prevention

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Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

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Section: Isolation and Characterization Of Crocin From C Sativusmentioning

confidence: 99%

Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation

Bakshi¹,

Hakkim²,

Sam³

2016

Asian Pacific Journal of Cancer Prevention

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“…The superna tant was frozen at -40°C for later use. The TAC of the saliva was determined by colorimetry, in accordance with the technique described by Lussignoli et al 9 The antioxidant power of the sam ple was characterised as the dose of the substance (or the volume of saliva) which caused a 50% inhibition of crocin bleaching (IC50). With the test sample we found it useful to express the TAC as the reciprocal of IC50 (TAC = 1/IC50) because in this way the TAC value is directly proportional (and not inversely proportional as in the case when IC50 is used) to the actual antioxidant power of the test sample.…”

Section: Evaluation Of Total Antioxidant Capacity (Tac)mentioning

confidence: 99%

Influence of the antioxidant content of saliva on dental caries in an at-risk community

et al. 2008

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• The amount of caries in deciduous teeth is in direct proportion to the observed antioxidant capacity of saliva.• The total antioxidant capacity of saliva is greater among children that have caries.• The presencce of caries in deciduous teeth is associated with caries in permanent teeth. I N B R I E F RESEARCHObjective This study aims to evaluate the relationship between the total antioxidant capacity of saliva and the presence of dental caries in deciduous and permanent teeth, in a group of Saharan children. Methods The dental examination was carried out in accordance with the recommendations of the World Health Organization (WHO). The total antioxidant capacity of the saliva was determined by colorimetry. Results The total antioxidant capacity (TAC) of the saliva of patients with caries in deciduous teeth was 2.89 1/IC50 greater than among those without. We observed a statistically signifi cant linear regression between the number of deciduous teeth affected by caries and the total antioxidant capacity of the saliva: y = 0.24 + 0.53 × TAC saliva (t = 2.93; p = 0.004) (95% CI of b: 0.018-0.088). Conclusions Our results show that the amount of caries in deciduous teeth is in direct proportion to the observed TAC of saliva, and that the presence of caries in deciduous teeth is associated with caries in permanent teeth.

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“…In this case, we could not monitor decreasing Abs under the photoirradiation. Instead, we evaluated the antioxidant activity as percent of inhibition relative to a control (%Inh) 33) using the ÁAbs value (eq. 5 and Table 2).…”

Section: )mentioning

confidence: 99%

Antioxidant Activity of Supramolecular Water-Soluble Fullerenes Evaluated by β-Carotene Bleaching Assay

Takada¹,

Kokubo²,

Matsubayashi³

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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A Microplate-Based Colorimetric Assay of the Total Peroxyl Radical Trapping Capability of Human Plasma

Cited by 72 publications

References 34 publications

Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation

Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation

Influence of the antioxidant content of saliva on dental caries in an at-risk community

Antioxidant Activity of Supramolecular Water-Soluble Fullerenes Evaluated by β-Carotene Bleaching Assay

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