Acridine Orange Propidium Iodide – Apoptosis of vascular smooth muscle cells induced by in vitro stimulation with interferon γ, tumor necrosis factor–α, and interleukin 1β, Comparing trypan blue & ao/pi staining methods, A family of basic amino acid transporters of the vacuolar membrane from saccharomyces cerevisiae*, Improved multi slice fourier ptychographic microscopy technique for high accuracy three dimensional tomography under oblique illuminations, View image, Anticancer activity of a monobenzyltin complex c1 against mda mb 231 cells through induction of apoptosis and inhibition of breast cancer stem cells
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Acridine Orange Propidium Iodide
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Potential Synergistic Activity Of Quercetin With Antibiotics Against Multidrug Resistant Clinical Strains Of Pseudomonas Aeruginosa
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Author: Landa Zeenelabdin Ali Salim 1, Syam Mohan 1, *, Rozana Othman 1, Siddig Ibrahim Abdelwahab 2, Behnam Kamalidehghan 1, Bassem Y. Sheikh 3 and Mohamed Yousif Ibrahim 1
There is a growing interest in naturally occurring compounds from traditional medicine that have anti-cancer potential. Nigella sativa (black seed) is one of the most studied plants. This annual herb grows in countries on the borders of the Mediterranean Sea and India. Timoquinone (TQ) is an active ingredient isolated from chicken sativa. The anti-cancer effect of TQ through the induction of apoptosis resulting from mitochondrial dysfunction was evaluated by IC in an acute lymphocyte leukemia cell line (CEMss).
1.5 ug / ml. A significant increase in chromatin condensation in the cell nucleus was observed by fluorescence analysis. Apoptosis was then confirmed by Annexin V, and an increasing number of cellular DNA fractures were observed in cells treated as a DNA ladder. Treatment of CEMss cells with TQ promoted apoptosis by signals that transmit cell death with low Bcl-2 regulation and high Bach regulation. In addition, significant cellular ROS, HSP70 generation, and activation of caspases 3 and 8 were observed in treated cells. Mitochondrial apoptosis was clearly associated with S-phase cell cycle arrest. Finally, the results of the current study showed that TQ may be a promising means of treating leukemia.
Antitumor‑ And Apoptosis‑inducing Effects Of Pomolic Acid Against Sk‑mel‑2 Human Malignant Melanoma Cells Are Mediated Via Inhibition Of Cell Migration And Sub‑g1 Cell Cycle Arrest Retraction In /10.3892/mmr.2022.12120
Also known as black rasca (black seed), cheriva sativa is a perennial herb of the Ranunculacea family that grows in countries bordering the Mediterranean Sea and India. It has been used as a traditional herbal medicine for more than 2,000 years. In addition to the use of this free drug, it is used as a medicinal food to promote health and fight many diseases in Asia, the Middle East and Africa . N. sativa is one of the most studied plants . It is used worldwide as a dietary supplement and aroma [2, 5]. They are also used as a natural remedy for asthma, hypertension, diabetes, inflammation, cough, bronchitis, headaches, eczema, fever, dizziness and influenza [4, 6]. The seeds are known to be carminative, stimulating, diuretic, emenagenous and galactologic and are used to treat fever . The biological activity of N. sativa seeds is associated with their essential oil components . The main compounds are thymokinone (30% -48%), p-simene (7% -15%), carvacrol (6% -12%), 4-terpineol (2% -7%), t-anethole (1). % –4% and sesquiterpene longifolene (1% –8%) .
In Malaysia, N. sativa is popularly known as “el Habbatus el Sauda”. At present, black raspberry oil is widely commercially available and from time to time numerous research projects are being carried out to improve its quality due to its amazing healing power [10, 11]. Thymokinone (TQ, 2-isopropyl-5-methyl-1,4-benzokinone, Figure 1b) has anti-inflammatory, antioxidant and anticarcinoma effects of this seed . In recent years, TQ and its effects on various cancer cell lines have been extensively studied; These effects include inhibition of cancer cell viability. Inhibition in almost all pancreatic cancer cell lines was up to 70% . Antiproliferative and proapoptotic activities of TQ in both NSCLC and SCLC cell lines . Breast cancer cell lines  and liver cell lines  have been studied in some animal models, including .
The burden of cancer is growing in Malaysia, as in other countries around the world, as it is one of the major health problems resulting in death . Leukemia is one of the most common childhood cancers in the world, as well as in Malaysia. Uterine leukemia accounts for 4.7% of childhood cancers in Malaysia . It is treated with multidisciplinary efforts, including chemotherapy. One of the main problems with current chemotherapy in the treatment of cancer patients is the toxicity of the drugs used. Unfortunately, most available anticancer drugs also attack normal proliferating cells . Therefore, there is currently a great interest in health for the consumption of natural compounds, which are generally considered safe. In the present study, we evaluated the potential of TQ in T-acute lymphoblastic leukemia using CEMss cells as an in vitro model.
The effect of TQ on the viability of CEMss cells was measured by MTT analysis. Cell proliferation after 24 hours of TQ exposure showed significant inhibition in TQ-treated cells compared to untreated cells (control). As shown in Figure 1, IC
The Z Stack Image Analysis Of Propidium Iodide (pi) And Acridine…
After 24 hours of treatment, the TQ was 1.5 ± 0.04 μg / ml. As the concentration of TQ increased, the proliferation of TQ-treated cells decreased.
Figure 1. Effect of thymokine on cell viability in CEMs. (a) Cell viability after 24 hours of treatment. Each point is averaged ± S.D. consists of three independent experiences. (b) Chemical structure of thymokine.
Apoptotic, necrotic and live CEMss cells were evaluated under a fluorescence microscope. This includes control cells (untreated); 200 cells were calculated randomly and differential. The study showed that the morphological features of TQ associated with apoptosis are triggered over time (Figure 2). Early apoptosis was evident with AO intercalated into fragmented DNA. In several such cases, the fluorescent bright green color can only be seen in processed CEMss cells. In contrast, untreated cells were observed to have a green solid nuclear structure. Blisters and nuclear marginalization (moderate apoptosis) were observed 24 hours after TQ treatment. In addition, late stages of apoptosis (i.e., the presence of an orange color due to AO binding to denatured DNA) were observed after 48 and 72 hours of TQ treatment (Figure 2). Differential evaluation of CEMss cells (200 cell population) treated as shown in Figure 3 showed a statistically significant difference (p 0.05) was found in the number of necrotic cells at different times (24, 48 and 72 hours) during treatment.
. (A, B) After 72 hours, untreated cells showed a normal structure without significant apoptosis and necrosis. (C, D) After 24 hours, signs of early apoptosis were observed, representing an orange (light green) stinging inserted between the fragmented DNA, (E, F) bleeding, and orange, which is a sign of late apoptosis after 48 hours. treatment, (G), H) bright red secondary necrosis occurred after 72 hours. VI: living cells; BL: swelling of the cell membrane; LA: late apoptosis; SN: secondary necrosis. The pictures represent one of three similar experiences.
Isothiocyanate From Moringa Oleifera Seeds Mitigates Hydrogen Peroxide Induced Cytotoxicity And Preserved Morphological Features Of Human Neuronal Cells
Figure 3. Percentage of viable, early apoptosis, late apoptosis and secondary necrotic cells after thymokinone treatment. Through apoptosis, CEMs cell death increased significantly over time (* p 0.05) in the number of necrosis cells was observed. VI: living cell, EA: early apoptosis, LA: late apoptosis, SN: secondary necrosis.
The induced apoptotic effect of TQ was further confirmed by determining the percentage of apoptotic cells by flow cytometric analysis with AV / PI double staining. AV + / PI_ staining represents early apoptotic cells due to the strong affinity of AV-FITC for phosphatidylserine from the inner layer of the plasma membrane to the outer surface of the membrane during early apoptosis. On the other hand, AV_ / PI + staining presents necrotic cells because PI, which cannot cross the entire cell membrane, penetrates the damaged membrane of dead cells or late apoptotic cells and binds to nucleic acid. At the same time, live cells can be labeled AV_ / PI_ and AV + / PI + staining is an indication of late apoptotic cells. Representative scatter plots of cytometric analysis of apoptosis flux showed that based on a comparison between untreated cells (control)
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