Irreversible Heat Inactivation of DNase I without RNA Degradation

Wiame, Ilse; Remy, Serge; Swennen, Rony; Sági, László

doi:10.2144/00292bm11

Cited by 93 publications

(44 citation statements)

References 5 publications

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“…RNA molecules are usually found stable in a pH range of 5.0 to 6.0. High pH (>7.5) and presence of divalent cataions favored the hydrolysis of phosphodiester bonds present in RNA (Wiame et al 2000). In the present method, EDTA was used to sequester bivalent cataions like Mn +2 or Mg +2 .…”

Section: Resultsmentioning

confidence: 99%

Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies

Rai¹,

Ghosh²,

Dey³

2010

Electron. J. Biotechnol.

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RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient, cost effective and reproducible RNA isolation method from hard tissue of bamboo internode. This protocol includes an additional organic solvent refinement steps to remove endogenous phenolic compounds and acidic phenol (pH 4.2) to critically stabilize RNA in extraction buffer. In addition to these, two 2M Lithium chloride washing steps were introduced to eliminate DNA and polysaccharides contamination. The RNA isolated from the present protocol was found to be superior, when compared to total RNA extracted by other available protocols. The A260/A280 absorption ratio of the isolated RNA was found ranging between 1.89-1.97. The integrity of 28S and 18S rRNA was highly satisfactory when analyzed in agarose denaturing gel. RNA was further used for RT PCR, northern hybridization, cDNA library and subtractive hybridization without any further refinement.

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Section: Resultsmentioning

confidence: 99%

Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies

Rai¹,

Ghosh²,

Dey³

2010

Electron. J. Biotechnol.

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“…The primers GPx (GPx-F: 5′-CTCTCCGCGGTGGCACAGT-3′, GPx-R: 5′-CCACCACCGGGTCGGACATAC-3) (GenBank: M21210). The primer Cu-Zn SOD (Cu-Zn SOD-F: 5′-GCAGAAGGCAAGCGGTGAAC-3′, Cu-Zn SOD-R: 5′-TAGCAGGACAGCAGATGAGT-3′) (GenBank: X05634) genes were normalized on the bases of GAPDH GAPDH -F: 5′-CAAGGTCATCCATGACAACTTTG-3′, GAPDH-R: 5′-GTCCACCACCCTGTTGCTGTAG -3′ (Wiame et al, 2000).…”

Section: Reverse Transcription and Semi-quantitative Polymerase Chainmentioning

confidence: 99%

Modulation of hepatotoxicity, DNA fragmentation and gene expression of Solanum nigrum leaves extract in rats treated with silver nanoparticles

2017

J App Pharma Sci

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The current study aimed to determine the antioxidant compounds in Solanum nigrum (S. nigrum) leaves extract, to synthesize silver nanoparticles (AgNPs) and to evaluate the protective role of the extract against the hepatotoxicity and genotoxicity of AgNPs compared to CCl4 in rats. Eight groups of female Sprague-Dawley rats were treated orally for 3 weeks included the control group, CCl4-trared group (0.1 ml/kg b.w twice a week), AgNPs-treated group (50 mg/kg b.w/day), AgNPs plus CCl4-treated group, S. nigrum leaves extract-treated group (0.5 mg/kg b.w) and the groups treated with AgNPs and/or CCl4 plus the extract. The results indicated that the extract was rich in the total phenolic, flavoniods and β-carotene. The size of synthesized AgNPs was 30-50 nm. Administration of AgNPs and/or CCl4 resulted in severe hepatotoxicity and histological changes, increased DNA fragmentation and down regulation of antioxidant gene expression in liver. The extract was safe and succeeded to mitigate the hazards effect of AgNPs and/or CCl4. It could be concluded that AgNPs have toxic effects and caution should be taken when they use in food or medical application. S. nigrum extract succeeded to protect the liver due to its higher content of antioxidant compounds.

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“…Cells were seeded in 6-well plate at 1 × 10 6 /mL cells and then incubated with the phenolic extracts as mentioned above. RNA extraction was assayed using Thermo Scientific Gene JET RNA Purification kit (Fermentas) according to the method described by Chomczynski and Sacchi [21], Boom [22], then first strand cDNA synthesis was assayed using RevertAid first strand cDNA synthesis kit (Fermentas) according to the method described by Wiame et al [23]. The reverse transcription reaction product can be directly used in polymerase chain reaction (PCR) applications.…”

Section: Apoptosis Assaymentioning

confidence: 99%

Anti-tumor effects of polyphenolic compounds of Camellia sinensis, Nigella sativa, Matricaria chamomilla and Triticum aesativum against Ehrlich ascites carcinoma mouse model

Salem¹,

Khamis²,

Mostafa³

et al. 2017

J Invest Biochem

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Background: Conventional anticancer chemotherapy is limited due to the associated with toxicity. Exploring new agents which can lower toxicity and enhance the efficacy of anticancer drug is of a paramount significance. Aim: Evaluate the antitumor activities of extracts of medicinal plants including green tea, chamomile, black cumin seeds, and wheat bran, as well as their capability to lower the toxicity induced by the conventional chemotherapeutic drugs cisplatin (CIS). Materials and Methods: The antitumor effects of these extracts were evaluated in in vitro and in vivo assays using Ehrlich ascites carcinoma (EAC) cells. In vitro studies, the cell viability and the expression of apoptosis-related genes Bad, Bax, Bcl-2, and Bcl-xL were analyzed by polymerase chain reaction technique. EAC cells were injected into mice followed by treatment with injection of CIS and oral administration of each extract or in combination. Results: The content of polyphenolic compounds in green tea, chamomile, black cumin, and wheat bran were 1200, 315, 35, and 20 mg/100 g of dry weight, respectively. Their 50% of inhibitory concentration values were 13.5, 30, 350, and 1060 mg/mL, respectively. The polyphenolic compounds suppressed EAC viability in vitro associated with up-regulation of Bad and Bax and down-regulation of Bcl-2 and Bcl-xL. Treatment of EAC tumor-bearing mice with these extracts associated with increases in the mean survival time of mice as well as increases and decreases in the numbers of dead and live EAC cells, respectively. The antitumor effect of a combinatorial treatment of the extracts together or with CIS was higher than those of single treatment. The hepatic and renal glutathione peroxidase and antioxidant capacity were significantly increased after treatment with the extracts. Conclusion: The extracts of these medicinal plants have potential antitumor effects when used in combination and can ameliorate the toxic effect of chemotherapy through antioxidant effects while enhance its antitumor effect through apoptotic effects.

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Irreversible Heat Inactivation of DNase I without RNA Degradation

Cited by 93 publications

References 5 publications

Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies

Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies

Modulation of hepatotoxicity, DNA fragmentation and gene expression of Solanum nigrum leaves extract in rats treated with silver nanoparticles

Anti-tumor effects of polyphenolic compounds of Camellia sinensis, Nigella sativa, Matricaria chamomilla and Triticum aesativum against Ehrlich ascites carcinoma mouse model

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