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Sexually Transmitted Diseases
Published in Victor A. Bernstam, Pocket Guide to GENE LEVEL DIAGNOSTICS in Clinical Practice, 2019
Fixation in Bouin solution markedly reduces the rate of detection of HPV DNA by ISH compared to that in tissue fixed with buffered formalin. Fixation and processing of tissues may impede the penetration of probes to the viral DNA.
Intercellular Communication in Three-Dimensional Culture
Published in Rolf Bjerkvig, Spheroid Culture in Cancer Research, 2017
Spheroids, heart fragments, and confrontation cultures were washed in PBS and fixed in Bouin solution for 2 h at room temperature. Cell cultures were then dehydrated in 70% ethanol for 2 to 3 h, 80 and 96% isopropanol for 30 min at room temperature, and in 100% isopropanol (replaced for three times) for 90 min at 45°C. After impregnation overnight at 57°C in a mixture of equal parts isopropanol and the embedding medium Paraplast Plus (Polyscience, Warrington, PA), cell cultures were infiltrated for 2 × 7 h at 57°C with pure Paraplast Plus and finally embedded in Paraplast Plus. For routine histology all specimens were cut to 5-µm-thick sections and stained with hematoxylin/erythrosine (Chroma, Köngen, FRG).
Atypical enteropathogenic E. coli are associated with disease activity in ulcerative colitis
Published in Gut Microbes, 2022
Maximilian Baumgartner, Rebecca Zirnbauer, Sabine Schlager, Daniel Mertens, Nikolaus Gasche, Barbara Sladek, Craig Herbold, Olga Bochkareva, Vera Emelianenko, Harald Vogelsang, Michaela Lang, Anton Klotz, Birgit Moik, Athanasios Makristathis, David Berry, Stefanie Dabsch, Vineeta Khare, Christoph Gasche
Fifty-seven E. coli isolates were grown on MacConkey agar for 24 hours under aerobic or anaerobic conditions, using Anaerobox and AnaeroGen sachets (Thermo Scientific, Oxoid). Single colonies were inoculated in 5 ml brain heart infusion (BHI, 37 g/L) medium with supplements (5 g/L yeast extract, 1 g/L NaHCO3, 1 g/L L-cysteine, 1 mg/L vitamin K1, 5 mg/L hemin) or in LB medium and grown under aerobic or anaerobic conditions for 6 hours at 37°C. Bacterial cells were diluted to an OD600 = 0.05. 100 µl of cell suspension was transferred to the U-bottom polystyrene 96-well plates (Costar) in four technical replicates. Plates were incubated at 37°C for 48 hours under aerobic or anaerobic conditions. Supernatants were removed, bacterial biofilms were fixed with 150 μL BOUIN solution (0.9% picric acid, 9% formaldehyde and 5% acetic acid) for 15 min and washed three times with 190 μL PBS. For staining, 150 μL 0.1% crystal violet solution was added for 10 min and washed three times with H20. For biofilm quantification, crystal violet in dried plates was dissolved in 190 μL 30% acetic acid and the plate was placed on a shaker for 1 h. Absorbance of 1:5 dilutions was measured on an Anthos 2010 microplate reader at 595 nm and 405 nm reference wavelength. Pairwise comparison was performed with the Mann–Whitney U test and ANOVA with Dunn’s multiple comparison test for comparing multiple groups.
Histological and biochemical investigation of the renoprotective effects of metformin in diabetic and prostate cancer model
Published in Toxicology Mechanisms and Methods, 2021
Pınar Koroglu-Aydın, Bertan Boran Bayrak, Ilknur Bugan, Omur Karabulut-Bulan, Refiye Yanardag
For the purpose of light microscope examinations, the kidney tissues were fixed for 22 hours in Bouin solution for routine histological procedure, processed routinely and embedded in paraffin. A 4 μm-thick sections taken at the microtome were stained using hematoxylin and eosin (HE). Semi-quantitative histological evaluation was made using an alteration of criteria defined by Fernandez et al. (2001). Each histological variable received a score between 0 (normal) and 3 (maximum damage). The scoring was done as none (−) 0, mild (+) 1, moderate (++) 2 and severe (+++) 3. These criteria were: observation of cell debris in the lumen of proximal and distal tubules, decrease in glomerular volume, presence of pycnotic nuclei, hyperemia, necrotic areas, hypertrophy and enlargement of the capsular area. Preparation samples were photographed with an Olympus CX41 microscope equipped with an Olympus DP71 digital camera DP7, Tokyo, Japan. A minimum of five area for each slide were analyzed and scored semi quantitatively for the seriousness of the changes by a blind observer.
Lobularia maritima leave extract, a nutraceutical agent with antioxidant activity, protects against CCl4-induced liver injury in mice
Published in Drug and Chemical Toxicology, 2022
Anis Ben Hsouna, Sabah Dhibi, Wissal Dhifi, Rania Ben Saad, Faical Brini, Najla Hfaidh, Jackson Roberto Guedes da Silva Almeida, Wissem Mnif
After fixation in Bouin solution, pieces of fixed tissue were embedded into paraffin, cut into 5 μm slices and colored with hematoxylin-eosin to examine tissue constitution (Gabe 1968). Six slices were prepared from each liver. All sections were evaluated semi-quantitatively for the degree of liver injury. The steatohepatitis calculation system was applied to evaluate necrosis, inflammation, and ballooning. Sirius red staining was also used to calculate the fibrotic area. In brief, the liver sections were incubated with picrosirius red for 2 h at room temperature and then washed with acetic acid and water. The percentage of the fibrotic area was calculated in 3 randomly selected fields per slide.