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1.
《Fungal biology》2020,124(2):83-90
Latterly, the upsurge in use of antifungal drugs has brought about the emergence of several drug-resistance strains, making it skeptical to continue relying on current therapeutic regime. In the necessity of resistance-free antifungal agent, flavonoids presented possibilities of replacing existing drugs, displaying antifungal activity against pathogenic fungi. Among them, quercetin, one of the most representative flavonoids, exhibited antifungal activity against Candida albicans. To inspect the further understanding regarding quercetin, the antifungal mode of action of quercetin was investigated. In the initial step, the apoptosis was monitored after quercetin treatment. Moreover, intracellular levels of Mg2+ was assessed and was determined that Mg2+ increase occurred under the influence of quercetin. In addition, several features of mitochondrial dysfunction were monitored. Mitochondrial dysfunction triggers decrease in mitochondrial redox levels and leads to disruption in mitochondrial antioxidant system. Increased intracellular ROS and decreased intracellular redox levels were also displayed, indicating the occurrence of overall disruption in antioxidant systems. Sequentially, DNA fragmentation was observed and this DNA damage in turn induces apoptosis. In analyses, hexaamminecobalt(III) chloride (Cohex) was applied to inhibit Mg2+ transport between cytosol and mitochondria. Cohex attenuated the effects induced by quercetin, which demonstrates that the presence of Mg2+ is essential in quercetin-induced apoptosis. 相似文献
2.
Viability measurements of hybridoma cells in suspension cultures 总被引:1,自引:0,他引:1
José M. Coco-Martin Jan W. Oberink Tiny A. M. van der Velden-de Groot E. Coen Beuvery 《Cytotechnology》1992,8(1):57-64
Several methods were applied to determine the viability of hybridoma cells in suspension. These methods include dye inclusion
and exclusion assays such as the classical trypan blue exclusion assay, the propidium iodide (PI) exclusion assay and the
fluorescein diacetate (FDA) inclusion assay. Furthermore, the relation was studied between release of lactate dehydrogenase
(LDH) by hybridoma cells and their viability. Also the ATP content of the cells and cellular heterogeneity as measured with
a flow cytometer were determined in relation to cellular viability.
The dye inclusion and exclusion assays using trypan blue, FDA, PI were shown to be useful methods to determine cellular viability.
With the FDA and PI methods it was possible to obtain additional information about cells which are in a transition state between
viable and non-viable. The viability according to the scatter properties of the cells appears to reflect the overall condition
of the cells, although interpretation of the results is difficult. Measurement of LDH release in the culture fluid or the
cytoplasmic ATP content could not be used as parameters for cell viability. 相似文献
3.
Scott L. Nyberg Russell A. Shatford William D. Payne Wei-Shou Hu Frank B. Cerra 《Biotechnic & histochemistry》1993,68(1):56-63
To establish the importance of fluorescein diacetate (FDA) as a viability stain for cultured hepatocytes. we hypothesized that FDA staining would correlate positively with hepatocyte viability and function. Mixtures of live and dead cells were stained with FDA and scanned by flow cytometry. A close correlation was observed between the live cell fraction and percent viability as determined by FDA staining (R2 = 0.962). Hepatocytes were also sorted into low fluorescence and high fluorescence groups. Both albumin production and lidocaine metabolism (P-450 activity) were significantly increased in the high fluorescence group compared to the low fluorescence group. An automated, fluorescence-activated assay was useful for rapid assessment of hepatocyte viability. In addition. the intensity of green fluorescence following staining with FDA correlated well with two specific measures of hepatocyte function. 相似文献
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肠道菌群在功能和代谢方面的研究日益成熟,但针对食品添加剂双乙酸钠对人体肠道菌群作用的研究目前仍鲜少有报道。为了探究其对肠道菌群及人体健康的潜在益处及危害,以小鼠为模型,以双乙酸钠为干预物质,干预剂量为0.3 g/(kg·d),干预1周后,收集实验小鼠和空白对照组小鼠的粪便并用菌群16S rDNA高通量测序进行肠道菌群的物种鉴定和丰度检测。对所得数据进行分析,得到小鼠肠道菌群的多样性与丰度比例等指标。结果显示,实验剂量下的双乙酸钠干预后,小鼠的肠道菌群无论是菌群种类还是丰度都有了显著改变,属水平新增447种,减少142种。根据丰度比例变化最显著的9种菌,如拟杆菌属(Bacteroides)、AKK(Akkermansia)等,并结合菌群代谢功能预测,脂肪酸合成显著上调,碳水化合物代谢、氨基酸代谢、脂代谢等明显下调,推测摄入高剂量的双乙酸钠可能有增加肥胖、过敏、慢性炎症和肠胃炎的风险。 相似文献
8.
Wenxiu Ye Mohammad Anowar Hossain Shintaro Munemasa Yoshimasa Nakamura Izumi C. Mori Yoshiyuki Murata 《Journal of plant physiology》2013
We recently demonstrated that endogenous abscisic acid (ABA) is involved in methyl jasmonate (MeJA)-induced stomatal closure in Arabidopsis thaliana. In this study, we investigated whether endogenous ABA is involved in MeJA-induced reactive oxygen species (ROS) and nitric oxide (NO) production and cytosolic alkalization in guard cells using an ABA-deficient Arabidopsis mutant, aba2-2, and an inhibitor of ABA biosynthesis, fluridon (FLU). The aba2-2 mutation impaired MeJA-induced ROS and NO production. FLU inhibited MeJA-induced ROS production in wild-type guard cells. Pretreatment with 0.1 μM ABA, which does not induce stomatal closure in the wild type, complemented the insensitivity to MeJA of the aba2-2 mutant. However, MeJA induced cytosolic alkalization in both wild-type and aba2-2 guard cells. These results suggest that endogenous ABA is involved in MeJA-induced ROS and NO production but not in MeJA-induced cytosolic alkalization in Arabidopsis guard cells. 相似文献
9.
Alleviation of chromium toxicity in rice seedlings by applying exogenous glutathione 总被引:1,自引:0,他引:1
Boyin Qiu Fanrong Zeng Shengguan Cai Xiaojian Wu Shamsi Imran Haider Feibo Wu Guoping Zhang 《Journal of plant physiology》2013
The effect of exogenous reduced glutathione (GSH) on alleviation of hexavalent chromium (Cr6+) toxicity to rice seedlings and its physiological mechanisms were comprehensively investigated in a series of experiments. Our results showed that growth and nutrient uptake of rice seedlings were dramatically reduced under 100 μM Cr6+ stress, and the reduction was significantly alleviated by exogenous GSH. Cr6+ stress also reduced cell viability in root tips and damaged ultrastructure of both chloroplasts and root cells, while the addition of GSH alleviates those negative effects. Cr-induced toxicity and GSH-caused Cr alleviation differed significantly between Cr-tolerant Line 117 (L117) and Cr-sensitive Line 41 (L41). Under Cr6+ stress, cystine content was increased and GSH content was decreased in rice plants, exogenous GSH, however, mitigated the Cr-toxicity by reversing the Cr-induced changes of the two compounds. The types of Cr-induced secretion of organic acids varied between the genotypes, where reduction in the contents of acetic and lactic acids and tartaric and malic acids were observed in L117 and L41, respectively. The addition of GSH alleviated the reduction of secretion of these organic acids. Exogenous GSH also altered the forms of Cr ions in the rhizosphere and the fraction of distribution at subcellular level in both shoots and roots. It may be concluded that the alleviation of Cr6+ toxicity by exogenous GSH is directly attributed to its regulation on forms of Cr ions in rhizosphere and their distribution at subcellular levels. 相似文献
10.
The free radical theory of aging emphasizes cumulative oxidative damage in the genome and intracellular proteins due to reactive oxygen species (ROS), which is a major cause for aging. Caloric restriction (CR) has been known as a representative treatment that prevents aging; however, its mechanism of action remains elusive. Here, we show that CR extends the chronological lifespan (CLS) of budding yeast by maintaining cellular energy levels. CR reduced the generation of total ROS and mitochondrial superoxide; however, CR did not reduce the oxidative damage in proteins and DNA. Subsequently, calorie-restricted yeast had higher mitochondrial membrane potential (MMP), and it sustained consistent ATP levels during the process of chronological aging. Our results suggest that CR extends the survival of the chronologically aged cells by improving the efficiency of energy metabolism for the maintenance of the ATP level rather than reducing the global oxidative damage of proteins and DNA. 相似文献