order Sulindac sulfide The significance of loop mediated iso

The significance of loop-mediated isothermal amplification (LAMP) method in microbial detection has been drawn increasing attentions recently [8]. It was developed by Notomi et al. in 2000 [9]. It amplifies nucleic acids within tens of minutes by self-circulation of strand displacement order Sulindac sulfide under isothermal conditions. With the advantages of high specificity and sensitivity, simplicity, rapidity and cheapness, it has been widely applied in many areas [10]. Research shows that invE gene exits in all Salmonella serotypes but not in non-Salmonella bacteria [11], while fliC, lygD and STM4495 specifically exists in S. Choleraesuis, S. Enteritidis and S. Typhimurium, respectively [12–14]. Therefore, the specific sequences in invE, fliC, lygD and STM4495 genes were selected as the target sequences for LAMP assay.
Materials and methods
Results and discussion
Conclusion In this study, we demonstrated that LAMP amplification for invE gene could successfully detected S. Enteritidis, S. Choleraesuis and S. Typhimurium with invE in their genome, while could not detect non-Salmonella strains. And LAMP amplifications for fliC, lygD and STM4495 genes could detect their corresponding Salmonella serotypes respectively, while did not cross-reacted with the irrelevant Salmonella serotypes or non-Salmonella strains. This indicated that the invE LAMP primers and the sequences in invE, fliC, lygD and STM4495 genes were highly specific for Salmonella. The detection limit of Salmonella was 2.0×101CFU/mL when LAMP assay was exploited for invE gene, which was 2.33×103CFU/mL by PCR for invE. The results indicated that LAMP assay was more sensitive than PCR. The LAMP reactions can be accomplished at a constant temperature of 63–65°C within 45–70min and terminated by inactivate the enzyme at 80°C for 10min. Just a thermostat water bath will work. Sample consumption is less and DNA template preparation is simple. Furthermore, identification methods of LAMP products are simple and various, such as observing precipitation by naked eyes, adding SYBR Green and agarose gel electrophoresis. In conclusion, the LAMP detection method for pathogenic Salmonella is specific, sensitive, simple, rapid and cost-effective. It has wide application prospect in the rapid detection of S. Choleraesuis, S. Enteritidis, S. Typhimurium and other pathogenic Salmonella in food.
Acknowledgments This work was supported by Wuhan Science and Technology Planning Project (201070934341), Wuhan Agricultural Technology Innovation Project (201120637175), and Wuhan Polytechnic University Major Incubation Planning (2011z01).
Introduction The link between oxidative stress, endothelial dysfunction and cardiovascular diseases (CVDs) has been well established [1]. Clinical studies have shown that there is increased production of reactive oxygen species in CVD patients as demonstrated by clinical studies [2]. The increase in reactive oxygen species induces oxidative stress, which then initiates atherosclerosis at the endothelium. Endothelial dysfunction is the first step in artherosclerosis and a major signal of cardiovascular complications [3]. Lee et al. [4] reported that oxidative stress is not only linked to endothelial dysfunction, but to all the processes involved in atherogenesis up to myocardial infarctions. Furthermore, hypercholesterolemia has been shown to induce oxidative stress [5] thereby causing tissue damage [6]. Lowering cholesterol levels has been shown to reduce the risk of myocardial infarctions and also increased endothelium dependent vasodilation [7]. A vasoconstrictor that has been linked to cardiovascular complications is angiotensin II, which is produced by the renin–angiotensin system and has also been linked to increased oxidative stress [8]. Therefore, the therapeutic targets for the management of cardiovascular complications include angiotensin-I converting enzyme (catalysing the conversion of angiotensin-I to angiotensin-II), 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (catalysing the conversion of HMG-CoA to mevalonate, which is the rate-limiting step in the synthesis of cholesterol and other isoprenoids) and the use of radical scavengers. The burden of cardiovascular diseases, which is a leading cause of death and disability in the world [9] is further increased by the cost of therapy which is beyond the reach of many in developing countries and the search for affordable therapy has led to natural alternatives. Grapefruits, like other citrus fruits have a smaller edible portion compared to the amount of non-edible portions such as peels and seeds. However, it has been shown that there are more bioactive compounds in the peels of citrus fruits than in the juices [10]. Grapefruit peels are domestically processed into candies and consumed by some Americans homes, while the peels are taken as infusion drinks in Asia. The bioactive compounds present in the peels, especially the phenolic compounds have been shown to have some medicinal properties [11]. However, these bioactive compounds in the peels have been of little benefit to humans as they are grossly under-utilized in many countries. This study sought to investigate the effect of phenolic extracts from grapefruit peels on key enzymes relevant to the management of cardiovascular diseases and also the cellular antioxidant activities of the extracts in endothelial cells.