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Essential Oils in Cancer Therapy
Published in K. Hüsnü Can Başer, Gerhard Buchbauer, Handbook of Essential Oils, 2020
Carmen Trummer, Gerhard Buchbauer
Tridax procumbens L. (Asteraceae) is a long-used traditional medical plant (Manjamalai et al., 2012). The following study showed that EO of T. procumbens L. is capable of suppressing lung metastasis by the B16F-10 cell line in C57BL/6 mice (20–25 g). In comparison with the untreated mice, the group which was treated with the EO showed an inhibition of tumor nodule formation of 71.7%. It was also found, that the EO was able to inhibit the formation of tumor directed new blood vessels (39.5%). The authors also studied apoptosis using the TUNEL assay. In comparison with cancer alone, the treated cells showed an increase of apoptotic cells (Manjamalai et al., 2012).
Role of Sperm DNA Damage in Male Infertility Assessment
Published in Botros Rizk, Ashok Agarwal, Edmund S. Sabanegh, Male Infertility in Reproductive Medicine, 2019
Saradha Baskaran, Chak-Lam Cho, Ashok Agarwal
TUNEL assay quantifies free DNA 3’-OH ends or nicks in the spermatozoa using fluorescent nucleotides [42]. Terminal deoxynucleotidyl transferase (TdT) incorporates fluorescent dUTP into 3’-OH groups of ssDNA or dsDNA breaks, which are quantified by fluorescence microscopy or flow cytometry. In the slide-based method, the sperm are categorized as TUNEL positive or negative, and results are expressed as percentage of total sperm in the population. In flow cytometry, the fluorescent signals are measured, which increases proportionally with the number of DNA strand breaks. The TUNEL assay is a simple, accurate, and reliable test for assessing DNA damage with low interobserver variation [43]. It measures both ss and dsDNA breaks. Recent study indicates that TUNEL coupled with flow cytometry increases the reproducibility and reliability of the test results and a cut-off value of 16.8% to discriminate among infertile and fertile men has been reported [44]. In addition, comparison of the data from identical semen samples across two reference laboratories (Basel, Switzerland, and Ohio, United States) revealed TUNEL as a robust test for measuring SDF in a multicenter setting [45].
Nerve Agent–Induced Seizures and Status Epilepticus: Neuroprotective Strategies
Published in Brian J. Lukey, James A. Romano, Salem Harry, Chemical Warfare Agents, 2019
Frederic Doreu, Karine Thibault, Nina Dupuis
Microglial cells can be detected using Griffonia simplicifolia (GSA) lectin staining (e.g., Pernot et al., 2011), but in some cases (several days after soman poisoning), we had difficulties using this technique due to very intense background staining, suggesting tissue changes (unpublished observations). The detection of microglia could then also be performed by immunohistochemistry of the ionized calcium binding adaptor molecule 1 (Iba1), a microglia/macrophage-specific calcium-binding protein (e.g., see Flannery et al., 2016). Astrocytes are usually located using immunohistochemistry of the glial fibrillary acidic protein (GFAP) (e.g. Baille-Le Crom et al., 1995; Flannery et al., 2016). Programmed cell death and apoptosis can also be studied by looking at specific proteins, such as caspases, p53, or Bax, or DNA damage (with the terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] assay, a method for detecting DNA fragmentation). It is worth mentioning that the TUNEL assay is not specific to apoptosis, and indeed, TUNEL-positive cells were showing other morphotypes of damage (Baille et al., 2005).
A novel polymerase β inhibitor from phage displayed peptide library augments the anti-tumour effects of temozolomide on colorectal cancer
Published in Journal of Chemotherapy, 2022
Lihong Qin, Mao Huiwen, Jianguo Wang, Yuanyaun Wang, Salman A. Khan, Ying Zhang, Hong Qiu, Longwei Jiang, Lingfeng He, Yan Zhang, Shaochang Jia
Moreover, we performed a terminal deoxytransferase-mediated dUTP biotin nick-end labelling (TUNEL) assay to examine cell apoptosis. Accumulation of unrepaired DNA damage will consequently activate the intrinsic pathway of apoptosis, which can be detected by TUNEL (TdT-mediated dUTP Nick-End Labeling) assay. TUNEL assay uses terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of deoxynucleotides at the free 3′-hydroxyl ends of fragmented DNA. The deoxynucleotides are then labelled for detection of the degree of DNA fragmentation. As shown in Figure 4C, the ratio of apoptotic cells were 2-fold higher in combination group than that of single-treated group. The representative micrograph of γH2AX foci detection and TUNEL assay were shown in Figure 4D,E respectively. The synergistic effect of TMZ and 10 D was analyzed with the isobologram method, as shown in Figure S3. These above results demonstrated that 10 D in combination with TMZ efficiently induce cytotoxicity in SW480 cancer cells.
Sperm DNA fragmentation on the day of fertilisation is not associated with assisted reproductive technique outcome independently of gamete quality
Published in Human Fertility, 2022
Jorge Ten, Jaime Guerrero, Ángel Linares, Adoración Rodríguez-Arnedo, Ruth Morales, Belén Lledó, Joaquín Llácer, Rafael Bernabeu
In relation to the sperm DNA fragmentation test employed, the SCSA and TUNEL methods are the most frequently used in published infertility studies (Cui et al., 2015). Nowadays, TUNEL assay is a routine diagnostic tool that measures DNA breaks by introducing fluorochrome labelled molecules. We decided to use TUNEL assay because this test measures DNA damage directly, without a denaturation step, such as the SCSA test (use of severely acid conditions) or Comet assay (use of alkaline conditions), and it is preferentially recommended (Panner-Selvam & Agarwal, 2018), showing the real fragmentation present in the DNA. In fact, the full nature of how these pH-exposures affect the sperm chromatin is not completely known at present (Barratt et al., 2010). Unfortunately, due to economic and technical reasons, flow cytometer was unavailable in our study and it was impossible to undertake with the TUNEL method.
Pristimerin inhibits neuronal inflammation and protects cognitive function in mice with sepsis-induced brain injuries by regulating PI3K/Akt signalling
Published in Pharmaceutical Biology, 2021
Weimin Xue, Yaqiang Li, Mei Zhang
The TUNEL assay was used to detect neuronal apoptosis. Paraffin-cleared brain slices were sealed with 0.1% (v/v) Triton X-100 at 37 °C for 15 min. After slices had been washed with phosphate-buffered saline, they were incubated with the TUNEL solution at 37 °C for 1 h in the dark. After slices had been washed with phosphate-buffered saline, they were incubated with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, St. Louis, MO) at 37 °C for 15 min. The slices were again washed with phosphate-buffered saline, then cover slipped prior to observation under an Olympus FV1000 confocal microscope (Olympus, Tokyo, Japan). TUNEL-positive neurons were counted in five random hippocampal regions. The apoptosis index was: (number of TUNEL-positive neurons/total number of DAPI-positive neurons)×100%.