Live/dead staining

  • 文章类型: Journal Article
    Sakazakii是婴儿配方奶粉(PIF)中的机会性食源性病原体,与几次食源性暴发有关。在PIF生产工厂中,他的生物膜可能是污染源,但是关于这些生物膜治疗的信息是有限的。本研究调查了在三个食品接触表面(不锈钢,硅胶,和PVC)使用高压大气冷等离子体(HVACP)在10:90空气:氦气改性的气氛中用作介质阻挡放电。在40kV的冷等离子体暴露90s后,与没有冷等离子体处理的情况相比,Sakazaki的CFU/优惠券减少了~3log。HVACP处理导致细胞收缩,细胞膜的碎片,和细胞质渗漏。通过活/死染色确认表面上细胞的失活。上述这些结果表明,HVACP对各种食品接触表面上的萨卡氏杆菌分离株的抗生物膜功效。
    Cronobacter sakazakii is an opportunistic foodborne pathogen in powdered infant formula (PIF) associated with several foodborne outbreaks. Biofilms of C. sakazakii in the PIF manufacturing plant may be a source of contamination, but information on the treatment of these biofilms is limited. This study investigated the inactivation of C. sakazakii biofilms on three food contact surfaces (stainless steel, silicone, and PVC) using high voltage atmospheric cold plasma (HVACP) applied as a dielectric barrier discharge in a 10:90 air: helium modified atmosphere. After the 90 s  of cold plasma exposure at 40 kV, C. sakazakii was reduced by ∼3 log CFU/coupon comparing to without cold plasma treatments. HVACP treatment caused cell shrinkage, fragmentation of the cell membrane, and leakage of cytoplasm. The inactivation of the cells on the surface was confirmed by live/dead staining. These above-mentioned results indicate the antibiofilm efficacy of HVACP on C. sakazakii isolates on various food contact surfaces.
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  • 文章类型: Journal Article
    研究的目的是研究实验改性牙科树脂复合材料表面初始生物膜的细菌活力。本研究包括25名口腔卫生良好的健康个体。在裂口设计中,他们收到了带有五个实验性复合树脂标本的丙烯酸夹板。其中四个用新型聚合物空心微珠递送系统或甲基丙烯酸酯化可聚合Irgasan(抗菌B)进行了修饰,而一个样本用作未修饰的对照(ST)。基于Poly-Pore®的递送系统装载有活性剂之一:Tego®Protect5000(抗粘连剂A),聚二甲基硅氧烷(防粘剂B),或Irgasan(抗菌A)。所有研究对象在研究期间避免刷牙。8小时后将样品从夹板上分离,并在荧光显微镜检查前给予活/死染色。应用Friedman检验和事后Nemenyi检验,显著性水平为p<0.05。总之,与ST相比,除抗菌B外的所有材料均显示出显着的抗菌作用。结果表明材料化学在细胞粘附的优势中的作用。总之,含有聚孔活性剂的牙科树脂复合材料显示出原位抗菌效果。
    The purpose of the study was to investigate the bacterial viability of the initial biofilm on the surface of experimental modified dental resin composites. Twenty-five healthy individuals with good oral hygiene were included in this study. In a split-mouth design, they received acrylic splints with five experimental composite resin specimens. Four of them were modified with either a novel polymeric hollow-bead delivery system or methacrylated polymerizable Irgasan (Antibacterial B), while one specimen served as an unmodified control (ST). A delivery system based on Poly-Pore® was loaded with one of the active agents: Tego® Protect 5000 (Antiadhesive A), Dimethicone (Antiadhesive B), or Irgasan (Antibacterial A). All study subjects refrained from toothbrushing during the study period. Specimens were detached from the splints after 8 h and given a live/dead staining before fluorescence microscopy. A Friedman test and a post hoc Nemenyi test were applied with a significance level at p < 0.05. In summary, all materials but Antibacterial B showed a significant antibacterial effect compared to ST. The results suggested the role of the materials\' chemistry in the dominance of cell adhesion. In conclusion, dental resin composites with Poly-Pore-loaded active agents showed antibacterial effectiveness in situ.
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  • 文章类型: Journal Article
    Studies on islet of Langerhans physiology are crucial to understand the role of the endocrine pancreas in diabetes pathogenesis and the development of new therapeutic approaches. However, so far most research addressing islet of Langerhans biology relies on islets obtained via enzymatic isolation from the pancreas, which is known to cause mechanical and chemical stress, thus having a major impact on islet cell physiology. To circumvent the limitations of islet isolation, we have pioneered a platform for the study of islet physiology using the pancreas tissue slice technique. This approach allows to explore the detailed three-dimensional morphology of intact pancreatic tissue at a cellular level and to investigate islet cell function under near-physiological conditions. The described procedure is less damaging and faster than alternative approaches and particularly advantageous for studying infiltrated and structurally damaged islets. Furthermore, pancreas tissue slices have proven valuable for acute studies of endocrine as well as exocrine cell physiology in their conserved natural environment. We here provide a detailed protocol for the preparation of mouse pancreas tissue slices, the assessment of slice viability, and the study of pancreas cell physiology by hormone secretion and immunofluorescence staining.
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