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Microbiology
Published in Karl H. Pang, Nadir I. Osman, James W.F. Catto, Christopher R. Chapple, Basic Urological Sciences, 2021
Katherine Belfield, Roger Bayston
Bacteria are divided into two groups based on gram stain (Hans Christian Gram method).Gram-positive bacteria: blue-black on staining (crystal violet stain).Gram-negative bacteria: red (safranin counterstain).
Cell Adhesion Molecules in Mast Cell Adhesion and Migration
Published in Bruce S. Bochner, Adhesion Molecules in Allergic Disease, 2020
Harissios Vliagoftis, Dean D. Metcalfe
Murine IL-3 dependent BMCMC are safranin-negative. When attached to fibroblasts through SCF for 2 weeks, they differentiate into safranin-positive cells. These cells also now synthesize heparin (6). If IL-3 is still present, these cells proliferate and produce more histamine than cells proliferating in the absence of adhesion to fibroblasts.
Animal Models of Osteoarthritis
Published in Yuehuei H. An, Richard J. Friedman, Animal Models in Orthopaedic Research, 2020
Theodore R. Oegema, Denise Visco
Most investigators use histopathology as an end point in their studies. However, one major limitation is an adequate validated grading scheme. Many grading schemes have been developed and used to describe the changes that occur in the articular cartilage observed in osteoarthritis,38,,108,109 and some have even been semi-automated.110 The most widely adopted grading method for evaluating microscopically hyaline cartilage changes was first described by Mankin et al. in 1971.109 However, the Mankin scheme has been described by one group to be reproducible,111 and by others to be inadequate.38,112 Furthermore, the use of the histochemical stain Safranin O has also been questioned by some researchers for its lack of reproducibility.113 Thus the need for a reproducible, standardized and validated grading scheme still exists and may involve scoring of area and severity of the surface fibrillation. For small animals, if no other analyses are planned, it may be possible to do this by scanning electron microscopy.14,19
Potential oral probiotic Lactobacillus pentosus MJM60383 inhibits Streptococcus mutans biofilm formation by inhibiting sucrose decomposition
Published in Journal of Oral Microbiology, 2023
Mingkun Gu, Joo-Hyung Cho, Joo-Won Suh, Jinhua Cheng
Biofilms were determined using safranin red assay [23]. For the co-culture model, both the overnight S. mutans and LAB culture were adjusted to OD600 of 1 and then diluted 100-fold with BHI containing 1% sucrose. For the treatment group, 20 µL of S. mutans and 20 µL of live LAB culture or heat-killed LAB culture were added to 160 µL of BHI medium containing 1% sucrose and incubated for 24 h at 37°C. After incubation. The biofilms in the 96-well plates were washed three times with distilled water and air dried. The biofilms were stained with 0.2% safranin red for 35 min at room temperature. After incubation, the dye was decanted from the wells and the biofilms were washed three times with distilled water without agitation, and then 125 μL of 33% acetic acid was added to extract the dye. The plates were shaken for 30 s before quantification of absorbance at 492 nm by a microtiter plate reader (Tecan, infinite M200PRO, Austria). For the control group, 20 µL of S. mutans and 20 µL of MRS broth were added to 160 µL of BHI medium containing 1% sucrose and incubated for 24 h at 37°C. This experiment was performed in triplicate.
Mitochondrial dysfunction: maternal protein restriction as a trigger of reactive species overproduction and brainstem energy failure in male offspring brainstem
Published in Nutritional Neuroscience, 2019
D. J. S. Ferreira, A. A. Pedroza, G. R. F. Braz, M. P. Fernandes, C. J. Lagranha
Several methods can be used to evaluate the membrane potential, herein we used the safranin-o uptake to estimate the electron difference between the mitochondrial spaces. In the assay, we evaluated the fluorescence decay, which has been correlated with the safranin attachment to internal mitochondria membrane in response to the increase in membrane potential. The animals that suffered protein restriction presented lower safranin-o uptake when compared to NP animals (NP= 35.61 ± 4.8 vs. LP= 19.32 ± 2.43% safranin uptake, P = 0.02) (Fig. 2a). Because ΔΨm can be regulated by UCP, which consumes proton motive force (pmf) without ATP formation, we evaluated the mRNA of UCP2. Our results demonstrated that UCP2 expression was expressed at higher levels in the brainstems of LP animals than in NP (NP = 0.063 ± 0.003 vs. LP = 0.204 ± 0.043, P = 0.03) (Fig. 2b).
An omentum-cultured 3D-printed artificial trachea: in vivo bioreactor
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2018
Hae Sang Park, Ji Seung Lee, Harry Jung, Do Yeon Kim, Sang Wook Kim, Md. Tipu Sultan, Chan Hum Park
The results of the histologic examination demonstrated that the tracheal graft was successfully integrated with the adjacent normal trachea in both groups (Figure 10). In the PCL-OC group, regenerated tissue on the luminal surface of the scaffold showed no definite inflammation and evidenced a highly organized structure composed of a ciliated respiratory epithelial layer and submucosal layer with numerous blood vessels at 4, 6 and 8 weeks postoperatively (Figure 10). The regenerated respiratory epithelial layer was composed of pseudocolumnar ciliated cells and goblet cells. Immunohistochemical staining showed a uniformly high expression of cytokeratin in the regenerated respiratory epithelial layer (Figure 11). This indicates mature epithelium lining the inner scaffold surface in the PCL-OC group. In contrast, regenerated tissue in the PCL group demonstrated severe inflammation and an unorganized structure (Figure 10). The PCL group showed partially regenerated ciliated cells at 4 weeks postoperatively, and the submucosal layer showed numerous inflammatory cells and fibrosis. Immunohistochemical staining against cytokeratin showed no definite positive cells (Figure 11). Figure 12 shows safranin O staining results in the PCL-OC group at 8 weeks. We could identify a small amount of neo-cartilage formation on the outer layer of the regenerated respiratory epithelium and submucosal layer.