China
Africa
Explore chapters and articles related to this topic
Radionuclide Concentrations in Soils lution-Processed Organic Solar Cells
Published in Michael Pöschl, Leo M. L. Nollet, Radionuclide Concentrations in Food and the Environment, 2006
Figure 5.6 shows a photograph of an untreated gray lowland sample (P38) of surface water after staining with SYBR Gold (a nucleic acid fluorescence dye). Most of the SYBR Gold-positive particles were characterized as spheres and rods; a few inorganic particles were also observed. Particles other than microbiological cells could be discriminated from cell particles by their shapes and fluorescence color. In addition to the inorganic particles, the numbers of protozoa and algae were also negligible. The microscopic observations indicated that the microbial components in the surface water sample were mainly fungi and bacteria. It was not possible to distinguish between fungi and bacteria by microscope observations, but the presence of fungal cells was confirmed by the formation of characteristic fungal colonies on an agar plate (data not shown).
Polypod-like structured guanine-rich oligonucleotide aptamer as a selective and cytotoxic nanostructured DNA to cancer cells
Published in Journal of Drug Targeting, 2021
Kohta Mohri, Emi Hayashi, Manato Nishino, Nao Matsushita, Sohei Tanishita, Makiya Nishikawa, Shinji Sakuma
As previously described, GRO and CRO assemblies were prepared by mixing equimolar amounts of corresponding ODNs [17]. Briefly, ODNs were dissolved in a buffer comprising 10 mM Tris-HCl, 1 mM ethylenediaminetetraacetic acid (EDTA) (TE buffer; pH 8) to obtain stock solutions of each ODN at a concentration of 0.1 mM. Then, ODNs were mixed and the concentrations of potassium chloride, sodium chloride and magnesium chloride were adjusted to 150, 150 and 50 mM, respectively. The mixtures were then incubated at 95 °C for 5 min, 65 °C for 2 min and 62 °C for 1 min and were then slowly cooled to 4 °C using a thermal cycler. The products were analysed by 8% or 20% polyacrylamide gel electrophoresis (PAGE) at 200 V for 40 min or 90 min, respectively. The gel was stained with SYBR Gold (Thermo Fisher Scientific, Inc., Waltham, MA) or ethidium bromide (EtBr) (Nippon gene, Tokyo, Japan) and observed using the GelDoc XR System with Quantity One 1-D analysis software (Bio-Rad Laboratories, Tokyo, Japan).
Human Amniotic Epithelial Cells Promote the Proliferation of Human Corneal Endothelial Cells by Regulating Telomerase Activity via the Wnt/β-catenin Pathway
Published in Current Eye Research, 2021
Jiayan Liu, Ye Wen, Wei Luo, Yingying Liu, Xiangyin Sha
Telomerase activity was detected using a TeloTAGGG Telomerase PCR ELISA Kit (Roche, Germany) as described previously.15 After 5 days of culture, HCEnCs were lysed for 30 min at 4°C. A 25 µl protein lysis solution and 25 µl of 2× TeloTAGGG reaction mix were mixed and incubated at room temperature for 20 min, which was followed by denaturation at 94°C for 5 min and then 33 cycles of the following: 94°C for 30 s, 50°C for 30 s, and 72°C for 90 s. Final elongation was carried out for 10 min at 72°C. Equal volumes of lysis solution, nuclease-free water and heat-inactivated 0.1% DMSO-treated samples (94°C for 5 min) were used as negative controls. PCR products (5 µl or 2.5 µl) were used for ELISAs according to the manufacturer’s protocol. Thirty microliters of the PCR products were loaded with 1× loading buffer on a TBE/12.5% polyacrylamide gel; then, electrophoresis was performed at 50 V for 30 min and a subsequent 180 V for 5–6 h. The gel was stained with 1× SYBR gold/TBE solution for 20 min and was visualised under UV light.
Bersaldegenin-1,3,5-orthoacetate induces caspase-independent cell death, DNA damage and cell cycle arrest in human cervical cancer HeLa cells
Published in Pharmaceutical Biology, 2021
Justyna Stefanowicz-Hajduk, Magdalena Gucwa, Barbara Moniuszko-Szajwaj, Anna Stochmal, Anna Kawiak, J. Renata Ochocka
The evaluation of DNA damage was analysed with the alkaline comet assay using the Reagent Kit for Single Cell Gel Electrophoresis Assay (Trevigen, Gaithersburg, MD, USA) according to the manufacturer’s instructions. HeLa cells were seeded at a density of 105 cells/well in 12-well plates. Cells were treated with bersaldegenin-1,3,5-orthoacetate at the concentrations of 1.0 and 5.0 µg/mL for 6 h and 24 h. Next cells were collected and combined with low melting agarose at a ratio of 1:10 and 50 µL of cell suspension was spread onto the CometSlides (Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA). Slides were immersed in the lysis solution for 1 h at 4 °C, after which they were placed in the alkaline unwinding solution for 1 h at 4 °C. Slides were next placed in the electrophoresis slide tray and immersed in the alkaline electrophoresis solution in the CometAssay ES unit (Trevigen). Slides were submitted to electrophoresis at 1 V/cm for 30 min. Following electrophoresis, slides were washed and then stained with Sybr Gold (Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA) for 30 min at RT. Slides were analysed under a fluorescence microscope (Nikon PCM-2000) and images of at least 50 comets per slide were analysed with the TriTek CometScore 2.0 software. Two regions of the comet were selected, the whole cellular DNA and the region of DNA within the comet head. The densities of the regions were defined and the results are presented as tail moment expressed as the percentage of DNA in the tail multiplied by the tail length. Each treatment was normalized to that of the control.