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Journal articles on the topic 'In vitro cytotoxic assays'
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1
Jurisic, Vladimir, and Vladimir Bumbasirevic. "In vitro assays for cell death determination." Archive of Oncology 16, no. 3-4 (2008): 49–54. http://dx.doi.org/10.2298/aoo0804049j.
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In this paper, we focused on commonly used in vitro assays for estimation of cell death: morphological analyses of cell death, cytotoxic assays based on enzymes activity determination, flow cytometry, and western blot techniques. We discussed advantages and disadvantages of several assays used in the modern research for estimation of cell death.
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Akca, Cetin, Ozgur Vatan, Dilek Yilmaz, Huzeyfe HURIYET, Nilüfer Cinkilic, and Tolga Cavas. "In vitro cytotoxic and genotoxic effects of donkey milk on lung cancer and normal cells lines." Czech Journal of Food Sciences 37, No. 1 (March 6, 2019): 29–35. http://dx.doi.org/10.17221/221/2018-cjfs.
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In vitro cytotoxic and genotoxic effects of donkey milk on cancer (A549) and normal (BEAS-2B) lung cell lines were investigated. The XTT and WST-1 tests as well as clonogenic assays were used to evaluate cytotoxicity. The comet assay and micronucleus test were used as genotoxicity endpoints. Donkey milk showed lower cytotoxic effects against normal lung cell line BEAS-2B in comparison to the tumor cell line A549. Genotoxicity experiments revealed dose dependent increases in the frequencies of micronuclei and single stranded DNA breaks in A549 cells whereas no significant damage was observed in BEAS-2B cells. The results indicate that donkey milk has anti-proliferative and genotoxic effects on lung cancer cells at concentrations which are non-toxic to normal lung cells.
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Lazaro, J. Enrico, and Frederick Gay. "Plasmodium falciparum: In Vitro Cytotoxicity Testing Using MTT." Journal of Biomolecular Screening 3, no. 1 (February 1998): 49–53. http://dx.doi.org/10.1177/108705719800300107.
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The microculture tetrazolium assay using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) was used to estimate the 50% inhibitory concentration of chloroquine, quinine, artemisinin, and atovaquone using a Plasmodium falciparum in vitro culture system. The MTT assay was compared to the standard tritiated hypoxan-thine assay and to a previously described method, the 2,2′-di-p-nitrophenyl-5,5′-diphenyl-3,3′-[3,3′-dimethoxy-4,4′-diphenylenel-ditetrazolium chloride (NBT) assay. In general, the results show that the three assays generate comparative results. The results of this study suggest that the MTT method is able to give a profile of cytotoxic dose response effects over a wide range of concentrations of a drug. The method may be used in work that does not require extreme pre-cision and sensitivity, for instance, as a portable rapid screen to assay natural products for in vitro cytotoxic ac-tivity against Plasmodium falciparum.
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Vencioneck Dutra, Jean, Jean Moisés Ferreira, Paula Costalonga Pereira, Judá Ben-Hur de Oliveira, Suiany Vitorino Gervásio, Mirieli Bernardes Xavier, Mainã Mantovanelli da Mota, et al. "Cereus jamacaru D.C. Hydroalcoholic Extract Promotes Anti-Cytotoxic and Antitumor Activity." Pharmaceuticals 11, no. 4 (November 23, 2018): 130. http://dx.doi.org/10.3390/ph11040130.
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Cereus jamacaru D.C. (mandacaru) is a cactus used as food and in the traditional medicine. In the present study, hydroalcoholic extract of C. jamacaru was evaluated for its chemical composition, antioxidant activity, cytotoxic and anti-cytotoxic effects in human lymphocytes and sarcoma 180 cells in vitro by MTT assay and antitumoral, mutagenic and cytotoxic effects on mice sarcoma-induced in vivo. Phytochemical characterization showed positive reactions for coumarin, flavanol and tyramine and total flavonoid content of 0.51 µg/mL. C. jamacaru showed antioxidant activity following DPPH (EC50 = 427.74 µg/mL), ABTS (EC50 = 270.57 µg/mL) and Fe2+ chelating ions assays (EC50 = 41.18 µg/mL). C. jamacaru induced significant decrease of sarcoma 180 viability at 24 h and 48 h of treatment, did not induce cytotoxicity in human lymphocytes and inhibits the cytotoxicity of cisplatin in vitro. Following in vivo assays, C. jamacaru promoted tumor reduction (86.07% of tumor inhibition), without inducing mutagenic or cytotoxic damage on mice blood cells. We propose that phenolic and alkaloid compounds in the extract are related to antioxidant activity, increasing its ability in metal chelating activity and promoting anti-cytotoxic activity against cisplatin, as well as these compounds may act on the cell cycle of the tumor cells in vitro and in vivo, leading to anticancer effects and tumor reduction.
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Ben Mrid, Reda, Najat Bouchmaa, Youssef Bouargalne, Btissam Ramdan, Khalid Karrouchi, Imad Kabach, Miloud El Karbane, Abderrazak Idir, Abdelmajid Zyad, and Mohamed Nhiri. "Phytochemical Characterization, Antioxidant and In Vitro Cytotoxic Activity Evaluation of Juniperus oxycedrus Subsp. oxycedrus Needles and Berries." Molecules 24, no. 3 (January 30, 2019): 502. http://dx.doi.org/10.3390/molecules24030502.
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In order to evaluate the antioxidant properties of aqueous and methanol extracts of needles and berries of Juniperus oxycedrus subsp. oxycedrus (Joo) species, various antioxidant capacity assessment tests (free radical scavenging assays (DPPH• and ABTS•+ tests), ferrous ions (Fe2+) chelating activity and reducing power assay (FRAP) were conducted. In all of the tests, the extracts exhibited strong antioxidant activity. Furthermore, in-vitro cytotoxic activity assays of the methanolic extracts showed potent cytotoxic effects against two breast cancer cell lines (MDA-MB-468 and MCF-7), with no cytotoxicity towards normal cells (PBMCs). Reactive oxygen species generation was presumed to be a potential reason for the observed cytotoxic effects. According to all the above, and considering its appropriate composition of mineral elements and phenolic compounds, Joo could offer a beneficial and natural source of bioactive compounds that can be either used on the preventive side as it could potentially be used in the clinic without toxicity.
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Güner, Adem. "In vitro risk assessment of Padina pavonica (Linnaeus) (Brown algae)." Food and Health 7, no. 1 (2021): 31–38. http://dx.doi.org/10.3153/fh21004.
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Padina pavonica (Linnaeus) Thivy 1960 is a brown algae that is antioxidant, antimicrobial, and anticancer effects and is generally used in soup, salad, and other dishes. However, no studies have been reported on safe consumption in humans to date. For this purpose, this study was conducted to determine the cytotoxic and genotoxic effects of P. pavonica on lymphocytes cultured from human blood. The water extract of P. pavonica was added into culture tubes at various concentrations (0.5-1000 μg/mL). Cytotoxic effects were determined by MTT assay. Antioxidant/oxidant status was evaluated by total antioxidant capacity (TAC) and total oxidative status (TOS) assays. Genotoxic effects were investigated by sister chromatid exchanges and micronucleus assays. Our results showed that P. pavonica had no genotoxic effects, even at higher concentrations. 1000 μg/mL concentration of P. pavonica caused an increase (P<0.05) TOS levels while significantly reducing cell viability. However, low concentrations (50 and 100 μg/mL) significantly increased (P<0.05) TAC levels. In conclusion, P. pavonica can be safely consumed with its non-genotoxic and antioxidant properties in a manner dose-dependent.
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Raajshree.r, Khoushika, and Brindha Durairaj. "IN VITRO ANTICANCER POTENTIAL OF BIOSYNTHESIZED ZINC OXIDE NANOPARTICLES FROM THE SEAWEED TURBINARIA CONOIDES." Asian Journal of Pharmaceutical and Clinical Research 11, no. 5 (May 1, 2018): 127. http://dx.doi.org/10.22159/ajpcr.2018.v11i5.22224.
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Objective: The objective of this study is to investigate the anticancer potential of zinc oxide nanoparticles (ZnO-NPs) and hydroethanolic extract of Turbinaria conoides (HETC) against Dalton’s lymphoma ascites (DLA) cell line. Methods: Nanoparticles were synthesized from the HETC. An ultraviolet-visible spectrophotometric analysis was performed to confirm the formation of ZnO-NPs. Size, morphology, and elemental composition of ZnO-NPs were also analyzed using scanning electron microscope-energy dispersive X-ray diffraction. The cytotoxic activity of ZnO-NPs and HETC was evaluated by 3-(4,5-dimethythiazol-. 2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue dye exclusion assays on DLA cells. The apoptosis inducing the effect was observed through acridine orange staining method (AO and EB) and DNA ladder assay. Results: The results of in vitro cytotoxic studies by MTT and trypan blue dye exclusion assays on DLA cell line in the presence of ZnO-NPs showed an IC50 value of 23.13 and 25.81 μg/ml, respectively. The DNA ladder assay and AO/EB staining clearly demonstrated that the ZnO-NPs at 50 μg/ml concentration induced a maximum apoptosis in DLA cells when compared with HETC. Conclusion: In the present study, the cytotoxic and apoptotic inducing effect of the synthesized ZnO-NPs and HETC were assessed, and it was found that ZnO-NPs possessed potent anticancer effect against DLA cells.
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Fukushima, Toshiro, Hitomi Tanaka, and Takeshi Yamamoto. "Comparative Study of Cigarette Smoke Cytotoxicity Using Two In Vitro Assay Systems." Beiträge zur Tabakforschung International/Contributions to Tobacco Research 26, no. 3 (September 1, 2014): 98–108. http://dx.doi.org/10.2478/cttr-2014-0013.
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SUMMARYThe aim of this study was to compare the results obtained from two in vitro cytotoxicity assays that depend upon different mechanisms/modes of action. The Neutral Red Uptake (NRU) assay is based on endocytotic activity whereas the Water Soluble Tetrazolium Salts (WST-1) assay is based on mitochondrial dehydrogenase activity. Both were investigated in light of their wide use and documented validation. The total particulate matter (TPM) and gas vapor phase (GVP) of main stream smoke derived from Kentucky reference cigarettes 3R4F and 10 test cigarettes made of 100% flue-cured or 100% Burley tobacco were individually applied to the two assays using CHO-K1 cells. In addition, cigarette smoke constituents and known cytotoxic agents, documented to affect specific endpoints, were evaluated within both assays. Although the NRU assay was primarily more sensitive than the WST-1 assay, both assays provided comparable results in terms of the rank order for the cytotoxicity of cigarette smoke samples. In addressing the cytotoxicity of constituents in cigarette smoke, acrolein, hydroquinone and catechol gave clear dose-related decreases in cell viability (an end point common in both assays). Moreover, enzyme inhibitors of the mitochondrial respiratory chain and chemicals causing membrane disruption also showed similar responses regardless of the specific endpoint addressed within the cytotoxicity assay. In conclusion, results from the NRU and WST-1 assay are comparable therefore indicating results were independent of the different assay detection mechanisms/modes of action. [Beitr. Tabakforsch. Int. 26 (2014) 98-108]
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Rodolfo, Monica, and Giorgio Parmiani. "Growth Inhibition of Murine Colonic Adenocarcinoma by Tumor Immune but not by IL-2-Activated or Alloactivated Lymphocytes." Tumori Journal 73, no. 1 (February 1987): 1–9. http://dx.doi.org/10.1177/030089168707300101.
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The antigenic profile of C-26 and C-51 BALB/c colonic adenocarcinomas was examined by in vivo and in vitro assays. Mice immunized with irradiated C-26 or C-51 tumor cells from freshly excised tumor nodules or from in vitro-growing cell lines were able to reject a challenge of both tumors. Spleen lymphocytes of immune but not of normal mice were effective in cross-inhibiting tumor growth in vivo in a Winn assay. Tissue-associated antigens common to C-26 and C-51 and to their metastases but not to other syngeneic neoplasms were detected in vitro by cytotoxic T lymphocytes obtained after 5 days of a secondary culture of immune lymphocytes and irradiated tumor cells. Activated lymphocytes were obtained by exposure of spleen cells to interleukin 2 or by allostimulation. Such lymphocytes, although cytotoxic in vitro on C-26 and C-51 carcinomas, were unable to significantly reduce in vivo tumor growth in the Winn assay.
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Levis, Mark, Patrick Brown, B. Douglas Smith, Adam Stine, Rosalyn Pham, Richard Stone, Daniel DeAngelo, et al. "Plasma inhibitory activity (PIA): a pharmacodynamic assay reveals insights into the basis for cytotoxic response to FLT3 inhibitors." Blood 108, no. 10 (November 15, 2006): 3477–83. http://dx.doi.org/10.1182/blood-2006-04-015743.
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Abstract We have developed a useful surrogate assay for monitoring the efficacy of FLT3 inhibition in patients treated with oral FLT3 inhibitors. The plasma inhibitory activity (PIA) for FLT3 correlates with clinical activity in patients treated with CEP-701 and PKC412. Using the PIA assay, along with in vitro phosphorylation and cytotoxicity assays in leukemia cells, we compared PKC412 and its metabolite, CGP52421, with CEP-701. While both drugs could effectively inhibit FLT3 in vitro, CEP-701 was more cytotoxic to primary samples at comparable levels of FLT3 inhibition. PKC412 appears to be more selective than CEP-701 and therefore less effective at inducing cytotoxicity in primary acute myeloid leukemia (AML) samples in vitro. However, the PKC412 metabolite CGP52421 is less selective than its parent compound, PKC412, and is more cytotoxic against primary blast samples at comparable levels of FLT3 inhibition. The plasma inhibitory activity assay represents a useful correlative tool in the development of small-molecule inhibitors. Our application of this assay has revealed that the metabolite CGP52421 may contribute a significant portion of the antileukemia activity observed in patients receiving oral PKC412. Additionally, our results suggest that nonselectivity may constitute an important component of the cytotoxic effect of FLT3 inhibitors in FLT3-mutant AML.
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Alqahtani, Ali S., Rashed N. Herqash, Omar M. Noman, Fahd A. Nasr, Nouf Alyhya, Shamsa H. Anazi, Muhammad Farooq, and Riaz Ullah. "In Vitro Antioxidant, Cytotoxic Activities, and Phenolic Profile of Senecio glaucus from Saudi Arabia." Evidence-Based Complementary and Alternative Medicine 2020 (October 24, 2020): 1–9. http://dx.doi.org/10.1155/2020/8875430.
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Current treatments for complex diseases have remarkable side effects that negatively impact patients’ quality of life. Thus, natural compounds with fewer side effects represent a promising source for safe drugs. The genus Senecio is widely used in folk medicine due to its various pharmacological properties. In the present study, the total phenolic content of Senecio glaucus, which is grown in Saudi Arabia, was assessed using the Folin-Ciocalteau colorimetric method. Scavenging DPPH and ABTS assays were utilized to determine the antioxidant properties of S. glaucus fractions, and MTT assay was used to screen the cytotoxic activity of S. glaucus against various cancer cells. In addition, HPLC-UV was utilized to detect the presence of two phenolic acids, namely, vanillic acid (VA) and gallic acid (GA). Among all fractions tested, S. glaucus chloroform fraction (SGCF) yielded the highest value (125.3 mg·GA/g) in terms of total phenolic content. SGCF also exhibited the highest scavenging activities (76.7 and 74.1%) on both DPPH and ABTS assays, respectively. Similarly, SGCF also possessed the most potent cytotoxic activity against the MCF-7 cell line, with an IC50 value of 41.8 μg/ml. The validated HPLC method confirmed the presence of VA (4.8 μg/mg DW) and GA (3.9 μg/mg DW) in SGCF. Overall, our data show that S. glaucus had antioxidant and cytotoxic properties. A developed validated HPLC method which could be helpful for quantifying phenolic compounds in S. glaucus was established.
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Bolla, Patricia Araceli, Paula Carasi, María de los Angeles Serradell, and Graciela Liliana De Antoni. "Kefir-isolatedLactococcus lactissubsp.lactisinhibits the cytotoxic effect ofClostridium difficile in vitro." Journal of Dairy Research 80, no. 1 (December 10, 2012): 96–102. http://dx.doi.org/10.1017/s0022029912000623.
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Kefir is a dairy product obtained by fermentation of milk with a complex microbial population and several health-promoting properties have been attributed to its consumption. In this work, we tested the ability of different kefir-isolated bacterial and yeast strains (Lactobacillus kefir, Lb. plantarum, Lactococcus lactissubps.lactis, Saccharomyces cerevisiaeandKluyveromyces marxianus) or a mixture of them (MM) to antagonise the cytopathic effect of toxins fromClostridium difficile(TcdA and TcdB). Cell detachment assays and F-actin network staining using Vero cell line were performed. Although incubation with microbial cells did not reduce the damage induced byC. difficilespent culture supernatant (SCS),Lc. lactisCIDCA 8221 and MM supernatants were able to inhibit the cytotoxicity of SCS to Vero cells. Fraction ofLc. lactisCIDCA 8221 supernatant containing components higher than 10 kDa were responsible for the inhibitory activity and heating of this fraction for 15 min at 100 °C completely abrogated this ability. By dot-blot assay with anti-TcdA or anti-TcdB antibodies, concentration of both toxins seems to be reduced in SCS treated withLc. lactisCIDCA 8221 supernatant. However, protective effect was not affected by treatment with proteases or proteases-inhibitors tested. In conclusion, we demonstrated that kefir-isolatedLc. lactisCIDCA 8221 secreted heat-sensitive products able to protect eukaryotic cells from cytopathic effect ofC. difficiletoxinsin vitro. Our findings provide new insights into the probiotic action of microorganisms isolated from kefir against virulence factors from intestinal pathogens.
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Silva, Claudia R. da, Marcia B. N. Oliveira, Ellen S. Motta, Gabriella S. de Almeida, Leandro L. Varanda, Marcelo de Pádula, Alvaro C. Leitão, and Adriano Caldeira-de-Araújo. "Genotoxic and Cytotoxic Safety Evaluation of Papain (Carica papayaL.) Using In Vitro Assays." Journal of Biomedicine and Biotechnology 2010 (2010): 1–8. http://dx.doi.org/10.1155/2010/197898.
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Papain, a phytotherapeutic agent, has been used in the treatment of eschars and as a debriding chemical agent to remove damaged or necrotic tissue of pressure ulcers and gangrene. Its benefits in these treatments are deemed effective, since more than 5000 patients, at the public university hospital at Rio de Janeiro, Brazil, have undergone papain treatment and presented satisfactory results. Despite its extensive use, there is little information about toxic and mutagenic properties of papain. This work evaluated the toxic and mutagenic potential of papain and its potential antioxidant activity against induced-H2O2oxidative stress inEscherichia colistrains. Cytotoxicity assay, Growth inhibition test, WP2-Mutoxitest and Plasmid-DNA treatment, and agarose gel electrophoresis were used to investigate if papain would present any toxic or mutagenic potential as well as if papain would display antioxidant properties. Papain exhibited negative results for all tests. This agent presented an activity protecting cells againstH2O2-induced mutagenesis.
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Hart, C. P., S. Ammons, J. X. Duan, D. Jung, J. Wang, H. Jiao, F. Meng, L. Lan, J. W. Evans, and M. Matteucci. "Discovery of TH-302: An achiral hypoxia-activated cytotoxic prodrug." Journal of Clinical Oncology 25, no. 18_suppl (June 20, 2007): 3515. http://dx.doi.org/10.1200/jco.2007.25.18_suppl.3515.
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3515 Background: Selective targeting of the hypoxic compartment of the tumor microenvironment is both a challenge and an opportunity for discovering and developing new cancer therapeutics. Chronic and transient hypoxia underlie resistance to chemotherapeutics and radiation. We have taken a modular design approach to synthesize hypoxia-activated prodrugs. Different nitro-containing heterocyclic triggers were linked to bis alkylating phosphorous mustard toxins and profiled in in vitro and in vivo assays predictive of compound efficacy and pharmacokinetics (PK). Methods: Drug candidates were synthesized and characterized using standard synthetic methods. In vitro efficacy was assessed with cancer cell line cytotoxicity and clonogenic assays. In vivo efficacy was assessed with xenograft models in monotherapy and combination therapy modes. Results: Screening trigger-toxin combinations in H460 lung cancer cell proliferation assays under anoxic, hypoxic, and aerobic conditions yielded structure-activity relationships (SAR) and identified TH- 302, a 2-nitroimidazole triggered bromo analog of ifosfamide, as a potent and selective molecule. TH-302 exhibits in vitro proliferation IC50 values of 0.2, 5, and 90μM and clonogenic IC90 values of 0.1, 5, and 30μM under conditions of 0%, 0.6%, and 21% O2, respectively. In vitro profiling showed sufficient solubility, plasma and microsomal stability, resistance to drug efflux mechanisms, and PK. In vivo efficacy was assessed with the H460 ectopic xenograft model. TH-302 administered alone (100mpk, ip, Q3Dx5) or in combination with cisplatin (6mpk, iv, Q7Dx2) showed mean tumor volume growth delays to 500mm3 (TGD500) of 10 days for TH-302 alone, 4 days for cisplatin alone, and 20 days for the combination. These results have been confirmed and extended using different tumor models, dosing regimens, and combinations which included Taxol and 5-FU. TH-302 demonstrates dose-proportional PK in multiple species. Conclusions: TH-302, a novel hypoxia-activated cytotoxic prodrug, has promising preclinical in vitro and in vivo efficacy and selectivity and has entered IND-enabling studies. Acute and subchronic toxicology is underway in rats and dogs. We expect to file an IND for clinical studies of TH-302 this year. No significant financial relationships to disclose.
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Chakrabarti, Srijita, Danswrang Goyary, Sanjeev Karmakar, and Pronobesh Chattopadhyay. "Exploration of cytotoxic and genotoxic endpoints following sub-chronic oral exposure to titanium dioxide nanoparticles." Toxicology and Industrial Health 35, no. 9 (September 2019): 577–92. http://dx.doi.org/10.1177/0748233719879611.
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Health hazards of titanium dioxide nanoparticles (TiO2-NPs) have raised severe concerns because of the paucity of information regarding the toxic effects among the population. In the present research, the in vitro and in vivo cytotoxic potential of TiO2-NPs were evaluated using flow cytometric techniques. Further, in vitro and in vivo genotoxic endpoints were estimated by means of comet, micronucleus (MN), and chromosomal aberration (CA) assays. In vitro analysis was performed at the concentration range of 10–100 µg/mL using murine RAW 264.7 cells. In vivo experiments were conducted on Albino mice (M/F) by exposing them to 200 and 500 mg/kg TiO2-NPs for 90 days. Decreased percentage of cell viability with higher doses of TiO2-NPs was evident in both in vitro and in vivo flow cytometric analysis. Further, an impaired cell cycle (G0/G1, S, and G2/M) was reflected in the present investigation following the exposure to TiO2-NPs. Increased comet scores such as tail length, % DNA in tail, tail moment, and olive moment were also observed with the higher doses of TiO2-NPs in vitro and in vivo comet assays. Finally, the in vivo MN and CA assays revealed the formation of MN and chromosomal breakage following the exposure to TiO2-NPs.
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Djordjevic, Dragana, Jelena Milovanovic, Milena Jurisevic, Bojana Stojanovic, Olga Cvetkovic, Marija Pergal, Elizabeta Ristanovic, et al. "Antitumour Effect of a Mixture of N-Propyl Polysulfides In Vitro." Serbian Journal of Experimental and Clinical Research 20, no. 4 (December 31, 2019): 295–300. http://dx.doi.org/10.1515/sjecr-2017-0069.
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Abstract Copper serves as a limiting factor for multiple steps of tumour progression, including angiogenesis, growth and metastasis. High levels of copper have been found in a wide spectrum of human cancers. Antitumour activities of copper-chelating drugs have been reported in animal models. Organosulfur compounds (diallyl sulfide, DAS; diallyl disulfide, DADS; S-ethylcysteine, SEC; N-acetylcysteine, NAC) derived from garlic exhibit marked copper-chelating activity. We analysed a mixture of fifteen n-propyl polysulfides (DPPS) for potential antitumour activity against several murine tumour cell lines, including colon carcinoma (CT26), mammary carcinoma (4T1) and melanoma cell lines (B16F10), and compared the effects with the antiproliferative effect in highly proliferative murine mesenchymal stem cells (mMSCs). The effects of the mixture of n-propyl polysulfides (100%) on cell viability were determined using MTT assays. Cell apoptosis was analysed using Annexin V-FITC/PI assays. The results of the MTT assays indicate that this standardized mixture of n-propyl polysulfides has a strong, dose-dependent cytotoxic effect against all three of the tested tumour cell lines (CT26, 4T1, B16F10). The cytotoxic effect of the n-propyl polysulfide mixture against the CT26 and B16F10 cell lines was much stronger than that of cisplatin and was significantly weaker in mMSCs, which are non-cancerous and highly proliferative cells, than in cancer cells. Flow cytometric analysis of CT26 and 4T1 cells revealed that apoptosis was not the dominant mechanism of cell death induced by the n-propyl polysulfide mixture. The n-propyl polysulfide mixture exerted highly cytotoxic activity against murine colon carcinoma and melanoma cell lines, but its antiproliferative activity against mMSCs was significantly lower than that of cisplatin.
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Languon, Sylvester, Isaac Tuffour, Emmanuel Ekow Quayson, Regina Appiah-Opong, and Osbourne Quaye. "In Vitro Evaluation of Cytotoxic Activities of Marketed Herbal Products in Ghana." Journal of Evidence-Based Integrative Medicine 23 (January 1, 2018): 2515690X1879072. http://dx.doi.org/10.1177/2515690x18790723.
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There are numerous herbal products on the Ghanaian market that are purported to cure various ailments, including cancer. However, scientific investigations on efficacy and toxicity of most of these products are not done. The aim of the study was to assess the anticancer potentials of herbal products on the Ghanaian market. Antiproliferative effects of Kantinka BA (K-BA), Kantinka Herbaltics (K-HER), Centre of Awareness (COA), a stomach (STO) and multicancer (MUT) product were evaluated in vitro using liver (Hep G2), breast (MCF-7), prostate (PC-3 and LNCaP), and blood (Jurkat) cancer cell lines. Cytotoxicity of the medicinal products was assessed using tetrazolium-based colorimetric assay, and total phenolic content and antioxidant activity of the products were determined using Folin-Ciocalteau and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays, respectively. Phytochemical screening resulted in the detection of terpenoids and flavonoids in most of the products, and alkaloids were detected in only MUT. Tannins were absent from all the products. The highest and lowest concentrations of phenolics were recorded for MUT and K-BA, respectively. The highest and lowest antioxidant activities were measured for MUT and K-HER, respectively. Only 2 products (STO and MUT) were cytotoxic to Hep G2 cells; with MUT being the only product that was cytotoxic to MCF-7 cells. All but K-BA were cytotoxic to PC-3 cells, while all products except K-HER were cytotoxic to LNCaP and Jurkat cells. The study thus confirms that the herbal products have selective cytotoxic activities against the tested cancer cell lines. However, comprehensive toxicity studies must be conducted to establish their safety.
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Pellissari, Claudia Viviane Guimarães, Carlos Eduardo Vergani, Elson Longo, Ana Claudia Pavarina, Paula Volpato Sanitá, Walter Luiz Siqueira, and Janaina Habib Jorge. "In Vitro Toxic Effect of Biomaterials Coated with Silver Tungstate or Silver Molybdate Microcrystals." Journal of Nanomaterials 2020 (January 28, 2020): 1–9. http://dx.doi.org/10.1155/2020/2971827.
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Purpose. This study evaluated the cytotoxicity of antimicrobial silver tungstate (Ag2WO4) or silver molybdate (Ag2MoO4) microcrystals coating biomaterials. Materials and Methods. The coating procedure was performed onto titanium, zirconia, and acrylic resin specimens. Eluates of the coated specimens were obtained, which were used for cytotoxicity analyses, including Alamar Blue, MTT, and CytoTox-ONE tests. Data were analyzed using two-way ANOVA, followed by the Tukey test (α = 0.05). The results of each experimental group were also compared to those of the control of living cells, taken as 100% cell viability. Results. In general, it was observed that the percentage of living cells from all biomaterials coated with both microcrystals was statistically different compared to the ones from the uncoated sample groups, except for the results from MTT of specimens of Ti coated with α-Ag2MoO4. All uncoated biomaterials were classified as noncytotoxic by the three assays used in the present study. It was observed that the microcrystals in solution were strongly cytotoxic, with death of almost 100% of cells, from the analysis of the results of the Alamar Blue assay. Conclusion. The most biomaterials coated with both microcrystals showed some degree of cytotoxicity in the different assays. The results described herein should be seen as an alert to the use of microcrystals, which can expose patients to health risks.
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Jadhav, Gajanan Raosaheb, and Priyankar Paira. "Cytotoxic 2‐(2′‐Hydroxyphenyl)benzothiazolylquinoline Analogues and their In Vitro Screening through Developmental Assays †." ChemistrySelect 4, no. 12 (March 28, 2019): 3524–30. http://dx.doi.org/10.1002/slct.201803618.
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Miret, Silvia, Els M. De Groene, and Werner Klaffke. "Comparison of In Vitro Assays of Cellular Toxicity in the Human Hepatic Cell Line HepG2." Journal of Biomolecular Screening 11, no. 2 (December 16, 2005): 184–93. http://dx.doi.org/10.1177/1087057105283787.
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Cytotoxicity testing allows determining whether a compound or extract contains significant quantities of biologically harmful chemicals. Cytotoxicity test methods are useful for screening because they serve to separate toxic from nontoxic materials, providing predictive evidence of compound safety. However, a wide range of assays measuring different aspects of cell death is available in the market, but it is difficult to determine which one(s) to use when evaluating a selection of compounds. The objective of this study was to compare different commercially available in vitro assays for cytotoxicity in HepG2 cells according to its sensitivity, reproducibility, simplicity, cost, and speed. The assays evaluated included Alamar Blue for the measurement of mitochondrial activity, ATPlite and ViaLight for the determination of cellular adenosine triphosphate (ATP), ToxiLight as an indicator of cellular necrosis, and Caspase-3 Fluorometric Assay, Apo-ONE Caspase-3/7 Homogeneous Assay, and Caspase-Glo for the determination of caspase-3/7 activity. All assays were performed using 4 compounds of previously reported cytotoxic activity: DMSO, butyric acid, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), and camptothecine. Overall, it was concluded that the best way to evaluate the potential cytotoxicity of a compound is to employ a battery of assays that focus on different aspects of cell death. In this case, the focus has been on ATP levels, cell necrosis, and capsase-3/7 activation. Many other kits are commercially available in the market for these and other aspects of necrosis and/or apoptosis. However, the use of ViaLight Plus, ToxiLight, and Caspase-3 Fluorometric Assay resulted in the most useful combination when working with HepG2 cells.
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Al-Ashaal, Hanan Abd Al-Hay. "Glycoalkaloids as medicinal agents from callus and regenerated plants of Solanum nigrum var. judaicum." Comunicata Scientiae 10, no. 3 (October 31, 2019): 325–37. http://dx.doi.org/10.14295/cs.v10i3.2840.
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The target of this study is production of glycoalkaloids from cultures of Solanum nigrum var. judaicum Besser. Further, to evaluate their therapeutic effects. S. nigrum var. judaicum leaves were implanted in MS media containing growth regulators for in vitro study. HPLC analyses were applied for qualitative and quantitative determination of glycoalkaloids. Cytotoxic effects against human carcinoma cell lines were evaluated. In addition, antiviral, antioxidant, anti-inflammatory and antiparasitic activities of the formed glycoalkaloids were estimated. HPLC data indicated the success of in vitro solasodine and solanidine glycosides production. Solasonine represented the highest concentration. Biological assays illustrated that obtained glycoalkaloids exhibited cytotoxic activity against human carcinoma cell lines that may be attributed to free radical scavenging activity (69.98%). Strong antiherps performance was observed (94%). In addition, the glycoalkaloids showed in vitro schistomicidal (IC50 76.4 ppm) and fasciolicidal (IC50 76.6 ppm) activities. In vivo anti-inflammatory assay revealed potent activity against carrageenan induced edema. Glycoalkaloids were formed 2-5 folds that of intact plant pointed to the efficiency of the cultures. The present findings referred to the pronounced biological performance of the produced glycoalkaloids including antiviral, cytotoxic anti-inflammatory and antiparasitic activities. Botanical derived medication from S. nigrum var. judaicum could be accomplished guided with the present data.
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Cannella, Vincenza, Roberta Altomare, Vincenza Leonardi, Laura Russotto, Santina Di Bella, Francesco Mira, and Annalisa Guercio. "In Vitro Biocompatibility Evaluation of Nine Dermal Fillers on L929 Cell Line." BioMed Research International 2020 (May 27, 2020): 1–6. http://dx.doi.org/10.1155/2020/8676343.
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Objective. Biomaterial research for soft tissue augmentation is an increasing topic in aesthetic medicine. Hyaluronic acid (HA) fillers are widely used for their low invasiveness and easy application to correct aesthetic defects or traumatic injuries. Some complications as acute or chronic inflammation can occur in patients following the injection. Biocompatibility assays are required for medical devices intended for human use, in order to prevent damages or injuries in the host. In this study, nine HA fillers were tested in order to evaluate their cytotoxicity and their effects on L929 cell line, according to the UNI EN ISO 10993 regulation. Methods. Extracts were prepared from nine HA fillers, and MTS viability assay was performed after 24 h, 48 h, and 72 h of exposure of cells to extracts. Cells cultured with HA filler extracts were monitored for up to 72 h, counted, and stained with haematoxylin/eosin in order to evaluate the cell proliferation rate and morphology. Results. None of the filler tested showed a cytotoxic effect. Two samples showed a higher vitality percentage and higher cell number while two samples showed a lower vitality percentage and lower cell number at 72 h. Conclusion. Data obtained suggest that although examined fillers are not cytotoxic, they show different effects on the in vitro cell proliferation rate. In vitro studies of medical devices could lead to important implications since these could aid to predict effects about their in vivo application. These easy and rapid assays could be useful to test new materials intended for human use avoiding animal tests.
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Wassmann, Torsten, Andrea Schubert, Felix Malinski, Martin Rosentritt, Sebastian Krohn, Kirsten Techmer, and Ralf Bürgers. "The antimicrobial and cytotoxic effects of a copper-loaded zinc oxide phosphate cement." Clinical Oral Investigations 24, no. 11 (March 20, 2020): 3899–909. http://dx.doi.org/10.1007/s00784-020-03257-w.
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Abstract Objectives Evidence about modifications of dental luting materials to minimize biological failure at the “marginal gap” between teeth and fixed prosthodontics is scarce. We compared a copper-modified (Co-ZOP) and a conventional zinc oxide phosphate cement (ZOP) in terms of antimicrobial and cytotoxic potentials in vitro and in vivo. Materials and methods Specimens of ZOP and Co-ZOP were characterized by the mean arithmetic roughness (Ra) and surface free energy (SFE). Powder components were examined using scanning electron microscopy (SEM). Energy-dispersive X-ray spectroscopy (EDX) showed elemental material compositions. In vitro microbial adhesion was shown using SEM, luminescence, and fluorescence assays. CCK-8 assays of mouse fibroblasts (L929) and human gingival fibroblasts (GF-1) were performed after 6, 24, and 48 h of specimen incubation. In vivo, ZOP and Co-ZOP specimens were applied intraorally for 12 h; biofilm accumulation was shown using SEM. Results Ra of ZOP and Co-ZOP showed no significant differences; SFE was significantly higher for Co-ZOP. EDX exhibited minor copper radiation for Co-ZOP, none for ZOP. In vitro fungal adhesion to Co-ZOP was significantly higher than to ZOP; in vitro streptococcal adhesion, cytotoxicity, and in vivo biofilm formation were not significantly different. Conclusions Co-ZOP showed low surface allocations of copper with no improved antimicrobial properties compared with conventional ZOP in vitro or in vivo. Clinical relevance Antimicrobial effects and low cytotoxicity of biomaterials are important for the clinical outcome. Based on our in vitro and in vivo results, no clinical recommendation can be given for the tested Co-ZOP.
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Ağalar, Hale Gamze, Gülşen Akalιn Çiftçi, Şafak Ulusoylar Yιldιrιm, Fatih Gögera, and Neşe Kιrιmera. "The LC/ESI-MSMS Profiles and Biological Potentials of Vitex agnus castus Extracts." Natural Product Communications 11, no. 11 (November 2016): 1934578X1601101. http://dx.doi.org/10.1177/1934578x1601101108.
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The chemical profile, cytotoxic and apoptotic effect, and antioxidant activity were determined of ethanolic extracts of Vitex agnus-castus L. (chaste tree). Ripened fruits and fruitless aerial parts were extracted with ethanol, and the chemical characterization of the extracts was determined by LC/ESI-MS-MS. Twelve compounds were tentatively identified in the extracts. The dose-dependent cytotoxic effects of the extracts were tested on C6, A549 and MCF-7 cells by using MTT assay; inhibition of DNA synthesis, and apoptotic and caspase-3 activation effects of the extracts were determined. The potential antioxidant activities of the extracts were evaluated by in vitro methods such as DPPH and ABTS scavenging activity, reducing power and β-carotene bleaching assays. The fruit extract showed noticeable cytotoxic activity against MCF-7 cells with an IC50 value of 88 μg/mL. Both extracts showed similar DPPH scavenging activity comparably with that of the standard.
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Omar, Nor Shamsuria, Thirumulu Ponnuraj Kannan, Abdul Rashid Ismail, Siti Fadilah Abdullah, Abdul Rani Samsudin, and Suzina Sheikh Abdul Hamid. "In Vitro Cytotoxic Evaluation of Processed Natural Coral in Human Osteoblasts." International Journal of Toxicology 30, no. 4 (May 3, 2011): 443–51. http://dx.doi.org/10.1177/1091581811399474.
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This study aimed to evaluate the in vitro cytotoxic effects of locally produced processed natural coral (PNC) using human osteoblasts (HOS). Cytotoxicity was not observed when HOS cells were cultured with PNC, as assessed by (3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide; MTT) and Neutral Red (NR) assays at concentration up 200 mg/mL for up to 72 hours. Flow cytometry (FCM) analysis showed that PNC (200 mg/mL) did not decrease viability of HOS cells after 48 and 72 hours of treatment. In a cell attachment study, the HOS cells attached to the edge of the PNC disc, and later grew into the pores of the PNC disc. All results from these studies indicate that locally produced PNC material is noncytotoxic and favors the growth of HOS cells.
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Rosa, S. S. Santa, F. O. Santos, H. G. Lima, I. M. A. Reis, D. S. A. Cassiano, I. J. C. Vieira, R. Braz-Filho, et al. "In vitro anthelmintic and cytotoxic activities of extracts of Persea willdenovii Kosterm (Lauraceae)." Journal of Helminthology 92, no. 6 (October 25, 2017): 674–80. http://dx.doi.org/10.1017/s0022149x17000979.
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AbstractThis study describes the effects of extracts and fractions of Persea willdenovii leaves against goat gastrointestinal nematodes and their cytotoxicity on Vero cells. The in vitro ovicidal and larvicidal activities of the crude ethanolic, hexane, ethyl acetate (EAE), butanolic and residual hydroethanolic extracts were assessed through the inhibition of egg hatching and larval motility assays. The most active extract (EAE) was then fractionated by chromatography in an open column containing silica gel, to furnish six fractions (Fr1–Fr6), which were also tested. The cytotoxicity of active extracts and fractions was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and trypan blue exclusion assay. The EAE and two fractions (Fr1 and Fr2) showed inhibitory activity in the egg hatching of gastrointestinal nematodes of goats in a concentration-dependent manner. The effective concentrations for 50% inhibition (EC50) of egg hatching were 2.3, 0.12 and 2.94 mg/ml for EAE, Fr1 and Fr2, respectively. All extracts and fractions were not effective in inhibiting 50% of motility of infective larvae. EAE and Fr2 had IC50 values (50% inhibitory concentration) of 4.95 and 2.66 mg/ml, respectively. Fr1 showed a slight cytotoxic effect (cellular inviability <30%) only after 48 h of treatment (MTT test). Gas chromatography–mass spectrometry (GC–MS) analysis showed the presence of six fatty acid ethyl esters, a fatty acid methyl ester and a long-chain ketone in the most active fraction. These constituents identified in P. willdenovii can be related to the high ovicidal activity and relatively non-toxic effect of the extracts.
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Gu, Qiang, Elvis Cuevas, Syed F. Ali, Merle G. Paule, Victor Krauthamer, Yvonne Jones, and Yongbin Zhang. "An Alternative In Vitro Method for Examining Nanoparticle-Induced Cytotoxicity." International Journal of Toxicology 38, no. 5 (June 24, 2019): 385–94. http://dx.doi.org/10.1177/1091581819859267.
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Conventional in vitro assays are often used as initial screens to identify potential toxic effects of nanoparticles (NPs). However, many NPs have shown interference with conventional in vitro assays, resulting in either false-positive or -negative outcomes. Here, we report an alternative method for the in vitro assessment of NP-induced cytotoxicity utilizing Fluoro-Jade C (FJ-C). To provide proof of concept and initial validation data, Ag-NPs and Au-NPs were tested in 3 different cell cultures including rat brain microvessel endothelial cells, mouse neural stem cells, and the human SH-SY5Y cell line. Conventional 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and lactate dehydrogenase (LDH) assays were run in parallel with the new method and served as references. The results demonstrate for the first time that FJ-C labeling can be a useful tool for assessing NP-induced cytotoxicity in vitro. Using these approaches, it was also demonstrated that removal of Ag-NPs—while keeping the Ag-ions that were released from the Ag-NPs in culture media—abolished the measured cytotoxicity, indicating that Ag-NPs rather than Ag-ions in solution contributed to the observed cytotoxic effects. Further, co-treatment of Ag-NPs with N-acetyl cysteine (NAC) prevented the observed cytotoxicity, suggesting a protective role of NAC in Ag-NP-induced cytotoxicity. Thus, this alternative in vitro assay is well suited for identify potential cytotoxicity associated with exposure to NPs.
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Otoguro, Kazuhiko, Kanki Komiyama, Satoshi Ωmura, and Charles A. Tyson. "An In Vitro Cytotoxicity Assay Using Rat Hepatocytes and MTT and Coomassie Blue Dye as Indicators." Alternatives to Laboratory Animals 19, no. 3 (July 1991): 352–60. http://dx.doi.org/10.1177/026119299101900309.
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Isolated hepatocytes from male Sprague-Dawley rats suspended in culture medium supplemented with either 0.2 or 2% bovine serum albumin (BSA) were allowed to attach to collagen coated 96-well dishes. Ten test chemicals from the MEIC list and salicylic acid were added individually to the dishes, and at the end of 24 and 48 hours, cytotoxicity was determined by measuring MTT (tetrazolium salt) reduction (mitochondrial integrity) and total cellular protein using Coomassie blue dye (reflecting cell number). Total cellular lactate dehydrogenase activity was also determined in some experiments, as an indicator of plasma membrane integrity. The relative toxicities of the test chemicals were quantified by the estimation of EC10, EC20 and EC50 values for each parameter. Except for one chemical, digoxin, in the MTT assay, cytotoxic potency increased with incubation time. The hepatocytes tended to be more sensitive to the chemicals in medium containing 0.2% BSA than in medium containing 2% BSA. Simple linear regression analyses of the log transformed data from the MTT assay versus log oral LD50 in rats for the test chemicals gave the best results using EC10 at 24 hours (r2 = 0.86). With protein as the cytotoxic indicator, the best results were obtained with EC values in the medium containing 2% BSA, again at 24 hours (r2 = 0.83). These results suggest that the MTT and Coomassie blue dye assays could be useful indicators for testing the cytotoxic potential of chemicals in rat hepatocyte cultures.
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Santos, P. A. S. R., G. B. Avanço, S. B. Nerilo, R. I. A. Marcelino, V. Janeiro, M. C. Valadares, and Miguel Machinski. "Assessment of Cytotoxic Activity of Rosemary (Rosmarinus officinalisL.), Turmeric (Curcuma longaL.), and Ginger (Zingiber officinaleR.) Essential Oils in Cervical Cancer Cells (HeLa)." Scientific World Journal 2016 (2016): 1–8. http://dx.doi.org/10.1155/2016/9273078.
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The objective of this study was to evaluate the cytotoxic activity of rosemary (REO,Rosmarinus officinalisL.), turmeric (CEO,Curcuma longaL.), and ginger (GEO,Zingiber officinaleR.) essential oils in HeLa cells. Cytotoxicity tests were performedin vitro, using tetrazolium (MTT) and neutral red assays for evaluation of antiproliferative activity by different mechanisms, trypan blue assay to assess cell viability and evaluation of cell morphology for Giemsa to observe the cell damage, and Annexin V to evaluate cell death by apoptosis. CEO and GEO exhibited potent cytotoxic activity against HeLa cells. IC50obtained was 36.6 μg/mL for CEO and 129.9 μg/mL for GEO. The morphology of HeLa cells showed condensation of chromatin, loss of cell membrane integrity with protrusions (blebs), and cell content leakage for cells treated with CEO and GEO, from the lowest concentrations studied, 32.81 μg/mL of CEO and 32.12 μg/mL of GEO. The Annexin V assay revealed a profile of cell death by apoptosis for both CEO and GEO. The results indicate cytotoxic activityin vitrofor CEO and GEO, suggesting potential use as anticancer agents for cervical cancer cells.
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Efstratiou, Androulla, Kathryn H. Engler, Charlotte S. Dawes, and Dorothea Sesardic. "Comparison of Phenotypic and Genotypic Methods for Detection of Diphtheria Toxin among Isolates of Pathogenic Corynebacteria." Journal of Clinical Microbiology 36, no. 11 (1998): 3173–77. http://dx.doi.org/10.1128/jcm.36.11.3173-3177.1998.
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We have compared molecular, immunochemical, and cytotoxic assays for the detection of diphtheria toxin from 55 isolates ofCorynebacterium diphtheriae and Corynebacterium ulcerans originally isolated in five different countries. The suitabilities and accuracies of these assays for the laboratory diagnosis of diphtheria were compared and evaluated against the “gold standard” in vivo methods. The in vivo and Vero cell cytotoxicity assays were accurate in their abilities to detect diphtheria toxin but were time-consuming; however, the cytotoxicity assay is a suitable in vitro alternative to the in vivo virulence test. There was complete concordance between all the phenotypic methods. Genotypic tests based upon PCR were rapid; however, PCR must be used with caution because some isolates of C. diphtheriae possessed toxin genes but failed to express a biologically active toxin. Therefore, phenotypic confirmation of toxigenicity for the microbiological diagnosis of diphtheria is recommended.
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Bahuguna, Ashutosh, Imran Khan, Vivek K. Bajpai, and Sun Chul Kang. "MTT assay to evaluate the cytotoxic potential of a drug." Bangladesh Journal of Pharmacology 12, no. 2 (April 8, 2017): 8. http://dx.doi.org/10.3329/bjp.v12i2.30892.
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<p>Quantification of cell viability and proliferation form the fundamental for numerous <em>in vitro</em> assays in response to external factors. An MTT assay is a colorimetric assay based on assessing the cell metabolic activity. A549 Lung adenocarcinoma cell line was used to see the cytotoxic potential of a new drug for initial screening of apoptosis or necrosis. The biochemical mechanism behind the MTT assay involves NAD(P)H-dependent cellular oxidoreductase enzyme that converts the yellow tetrazolium MTT [3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] into insoluble (E,Z)-5-(4,5-dimethylthiazol-2-yl)-1,3-diphenylformazan (formazan). The formed formazan can be dissolved with dimethyl sulfoxide (DMSO) to give a purple color with characteristic absorption at 540 nm. Intensity of purple color is directly proportional to the cell number and thus indicating the cell viability.</p><p><strong>Video Clip of Methodology:</strong> 3 min 56 sec <a href="https://www.youtube.com//v/eqFxzDVunt8">Full screen</a> <a href="https://www.youtube.com/watch?v=eqFxzDVunt8">If Failed</a></p>
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Mahmoud, Abudayyak, Öztaş Ezgi, Arici Merve, and Gül Özhan. "In Vitro Toxicological Assessment of Magnesium Oxide Nanoparticle Exposure in Several Mammalian Cell Types." International Journal of Toxicology 35, no. 4 (May 13, 2016): 429–37. http://dx.doi.org/10.1177/1091581816648624.
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Worldwide researchers have rising concerns about magnesium-based materials, especially magnesium oxide (MgO) nanaoparticles, due to increasing usage as promising structural materials in various fields including cancer treatment. However, there is a serious lack of information about their toxicity at the cellular and molecular levels. In this study, the toxic potentials of MgO nanoparticles were investigated on liver (HepG2), kidney (NRK-52E), intestine (Caco-2), and lung (A549) cell lines. For the toxicological assessment, the following assays were used: the particle characterization by transmission electron microscopy, the determination of cellular uptake by inductively coupled plasma-mass spectrometry, MTT and neutral red uptake assays for cytotoxicity, comet assay for genotoxicity, and the determination of malondialdehyde (MDA), 8-hydroxydeoxyguanosine, protein carbonyl, and glutathione levels by enzyme-linked immune sorbent assays for the potential of oxidative damage and annexin V-fluorescein isothiocyanate (FITC) apoptosis detection assay with propidium iodide (PI) for apoptosis. Magnesium oxide nanoparticles were taken up by the cells depending on their concentration and agglomeration/aggregation potentials. Magnesium oxide nanoparticles induced DNA (≤14.27 fold) and oxidative damage. At a concentration of ≥323.39 µg/mL, MgO nanoparticles caused 50% inhibition in cell viability by 2 different cytotoxicity assays. The cell sensitivity to cytotoxic and genotoxic damage induced by MgO nanoparticles was ranked as HepG2 < A549 < Caco-2 < NRK-52E. Although it was observed that MgO nanoparticles induced apoptotic effects on the cells, apoptosis was not the main cell death. DNA damage, cell death, and oxidative damage effects of MgO nanoparticles should raise concern about the safety associated with their applications in consumer products.
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Karaaslan, Cigdem, Hande Gurer-Orhan, Sibel Suzen, Luciano Saso, Omidreza Firuzi, Marjan Tavakkoli, and Elif Ince. "Behaviour of 9-Ethyl-9H-carbazole Hydrazone Derivatives Against Oxidant Systems." Croatica chemica acta 92, no. 1 (2019): 87–94. http://dx.doi.org/10.5562/cca3481.
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Antioxidants are helpful in prevention of several diseases related with oxidative stress including neurodegenerative disorders. In recent studies, carbazoles were given proof of promising antioxidant activities. In this article, 9-ethyl-9H-carbazole hydrazone derivatives were synthesized, characterized and their in vitro antioxidant activity and possible cytotoxic effects were investigated. Furthermore, protective effect of the synthesized derivatives against amyloid β-induced damage in PC12 neuronal cells was examined by using MTT assay. The newly synthesized carbazoles were found to have radical scavenging activity with a varying potency both in cell-free and cell-based in vitro assays. Several compounds, especially such as 3d and 3e, 3m and 3n bearing two halogen groups on the phenyl ring, were found to have cytotoxic activity. However, their cytotoxic activities were not higher than that of melatonin. Several compounds also significantly protected neuronal PC12 cells against amyloid β-induced damage, which can be defined as neuroprotective agents. (4-(2-((9-Ethyl-9H-carbazol-3-yl)methylene)hydrazinyl)benzonitrile) 3r was found as the most active compound with both radical scavenging activity and neuroprotective effects against amyloid β-induced damage. These findings might provide an alternative strategy for developing novel carbazole derivatives for management of neurodegenerative diseases, such as Alzheimer's disease.
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Friedman, Daphne R., Yvonne Mowery, Margaret Kennedy, Karen M. Bond, J. Brice Weinberg, and George J. Cianciolo. "The Anti-Inflammatory Investigational Agent LMP-420 Demonstrates in Vitro Cytotoxic Activity against Chronic Lymphocytic Leukemia Cells." Blood 114, no. 22 (November 20, 2009): 2358. http://dx.doi.org/10.1182/blood.v114.22.2358.2358.
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Abstract Abstract 2358 Poster Board II-335 Background: Chronic Lymphocytic Leukemia (CLL) is a common incurable hematologic malignancy. When therapy is required, maximizing durable responses is often at the risk of increasing toxicity. Thus, developing novel therapeutic agents that have minimal overlapping toxicity with currently used chemotherapy would be advantageous. To this end, we investigated LMP-420, a boronic acid containing purine nucleoside analogue, that potently inhibits tumor necrosis factor alpha (TNF) transcription in stimulated peripheral blood mononuclear cells (PBMCs) without affecting cell viability. Since TNF has been implicated in promoting CLL cell viability and can be produced by CLL cells themselves, we hypothesized that LMP-420 would be cytotoxic for CLL lymphocytes, either alone or in combination with fludarabine. Methods: To test the activity of LMP-420, we negatively selected circulating CLL cells from blood collected from patients using the RosetteSep B-cell enrichment cocktail (StemCell Technologies) and a Ficoll-Hypaque gradient. This method yields greater than 95% purity of malignant lymphocytes, determined by immunophenotyping CD5+CD19+ cells. Prognostic markers such as IgVH mutation status, CD38 and ZAP70 expression, and interphase cytogenetics were determined as previously described. We assessed the fractional toxicity and 50% effective dose (ED50) of LMP-420, fludarabine, or the combination, with the MTS colorimetric cytotoxicity assay, in which CLL cells were incubated for three days in Hybridoma media + 10% fetal bovine serum with serial dilutions of drug alone or in combination. Apoptosis was measured via annexin V flow cytometry methods and caspase 3/7 activity assays. We determined the effect of LMP-420 compared to fludarabine on the normal hematopoietic system by testing serial dilutions of both agents in killing normal PBMCs and in suppressing erythroid and myeloid colony formation. Results: The median ED50 of LMP-420 for CLL cells was 423 nM (range 0.01 to 2224 nM, n = 21). Two patients had high-risk cytogenetics (17p or 11q deletions), and their ED50 values for LMP-420 were 691 and 90 nM, respectively. The cytotoxic effect of fludarabine was potentiated on average 80 or 261 fold with the addition of LMP-420 at concentrations of 62 or 250 nM, respectively (ranges 1.14 – 947 and 1.19 – 2754). This agent killed malignant lymphocytes by apoptotic mechanisms in a dose-responsive fashion, as demonstrated by both Annexin V staining and caspase 3/7 activity assays. While LMP-420 has potent anti-CLL activity, it has minimal effects on normal hematologic cells. For example, fludarabine suppresses erythroid and myeloid colony formation by greater than 50% at a concentration of 1 uM, while this level of inhibition is seen for LMP-420 at a concentration of 90 uM. The average cytotoxic ED50 of LMP-420 on normal PBMCs using the MTS assay was greater than 90 uM, whereas for fludarabine, it was 5.3 uM. This finding was confirmed with apoptosis assays. Conclusions: The results of these experiments demonstrate that LMP-420, a novel inhibitor of TNF expression, has cytotoxic activity against CLL cells, including those with high-risk features. LMP-420 appears to increase the cytotoxic effect of the chemotherapy agent fludarabine, while imparting minimal increase in hematologic toxicity. Thus, LMP-420 is promising new therapeutic agent in CLL. Disclosures: No relevant conflicts of interest to declare.
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Takemoto, Jody K., Connie M. Remsberg, and Neal M. Davies. "Pharmacologic Activities¬ of 3’-Hydroxypterostilbene: Cytotoxic, Anti-Oxidant, Anti-Adipogenic, Anti-Inflammatory, Histone Deacetylase and Sirtuin 1 Inhibitory Activity." Journal of Pharmacy & Pharmaceutical Sciences 18, no. 4 (November 10, 2015): 713. http://dx.doi.org/10.18433/j33w4c.
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Purpose: Delineate the selected pharmacodynamics of a naturally occurring stilbene 3’-Hydroxypterostilbene. Objective: Characterize for the first time the pharmacodynamics bioactivity in several in-vitro assays with relevant roles in heart disease, inflammation, cancer, and diabetes etiology and pathophysiology. Methods: 3’-Hydroxypterostilbene was studied in in-vitro assays to identify possible bioactivity. Results: 3’-Hydroxypterostilbene demonstrated anti-oxidant, anti-inflammatory, cytotoxic, anti-adipogenic, histone deacetylase, and sirtuin-1 inhibitory activity. Conclusions: The importance of understanding individual stilbene pharmacologic activities were delineated. Small changes in chemical structure of stilbene compounds result in significant pharmacodynamic differences. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.
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Choudhury, Aniruddha, James L. Gajewski, Jan C. Liang, Uday Popat, David F. Claxton, Kay-Oliver Kliche, Michael Andreeff, and Richard E. Champlin. "Use of Leukemic Dendritic Cells for the Generation of Antileukemic Cellular Cytotoxicity Against Philadelphia Chromosome-Positive Chronic Myelogenous Leukemia." Blood 89, no. 4 (February 15, 1997): 1133–42. http://dx.doi.org/10.1182/blood.v89.4.1133.
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Abstract The success of adoptive immunotherapy for the treatment of leukemia depends on the generation of T cells that can specifically react with malignant cells. Dendritic cells (DCs) are important antigen-presenting cells in the development of antileukemic T-cell responses. In this study, we generated DCs from peripheral blood cells of patients with chronic myelogenous leukemia (CML). CML cells incubated concurrently with granulocyte-macrophage colony-stimulating factor, interleukin-4, and tumor necrosis factor-α in vitro developed morphologic and phenotypic characteristics of DCs. Fluorescence in situ hybridization showed the presence of t(9; 22) in the nuclei of these cells, indicating that they were leukemic in origin. These cells were potent stimulators of lymphocyte proliferation in specific in vitro assays for DC function. Autologous T cells stimulated with in vitro-generated, leukemic DCs displayed vigorous cytotoxic activity against CML cells but low reactivity to major histocompatability complex-matched normal bone marrow cells. Cytotoxic activity against CML targets was fourfold to sixfold higher using DC-stimulated autologous T cells than with autologous T cells expanded by culture with interleukin-2 alone. DC-stimulated T cells also inhibited growth of CML clonogenic precursors in colony-forming assays in vitro. These results suggest that cytokine-driven in vitro differentiation of CML cells results in generation of DCs with potent T-cell stimulatory function. In vitro-generated DCs can be effectively used as antigen-presenting cells for the ex vivo expansion of antileukemic T cells.
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Seroka, Barbara, Zenon Łotowski, Agnieszka Hryniewicka, Lucie Rárová, Rafal R. Sicinski, Aneta M. Tomkiel, and Jacek W. Morzycki. "Synthesis of New Cisplatin Derivatives from Bile Acids." Molecules 25, no. 3 (February 4, 2020): 655. http://dx.doi.org/10.3390/molecules25030655.
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A series of bile acid derived 1,2- and 1,3-diamines as well as their platinum(II) complexes were designed and synthesized in hope to get a highly cytotoxic compound by the combination of two bioactive moieties. All complexes obtained were subjected to cytotoxicity assays in vitro and some hybrid molecules showed an expected activity.
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Hunt, Stephen M., Christina Chrzanowska, Christopher R. Barnett, Helen N. Brand, and John K. Fawell. "A Comparison of In Vitro Cytotoxicity Assays and Their Application to Water Samples." Alternatives to Laboratory Animals 15, no. 1 (September 1987): 20–29. http://dx.doi.org/10.1177/026119298701500104.
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A group of 13 compounds were tested for in vitro cytotoxicity in four test systems; MIT-24 test, inhibition of cell growth (protein method), inhibition of cell growth (vital dye method) and cloning efficiency. In general, all four assays tended to rank compounds in a similar order for toxicity. The length of the exposure period appeared to be important for some compounds. The cytotoxicity of a variety of water samples was examined in two tests; inhibition of cell growth (vital dye method) and cloning efficiency. Under the conditions in which the assays were carried out, the latter proved to be the more sensitive test. River water samples gave little or no indication of cytotoxicity, samples of domestic sewage effluent gave some evidence of cytotoxicity, while an industrial effluent was markedly cytotoxic.
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Das, Swagat K., Sagarika Dash, Hrudayanath Thatoi, and Jayanta K. Patra. "In vitro α-amylase and α-glucosidase Inhibition, Antioxidant, Anti- Inflammatory Activity and GC-MS Profiling of Avicennia alba Blume." Combinatorial Chemistry & High Throughput Screening 23, no. 9 (December 22, 2020): 945–54. http://dx.doi.org/10.2174/1386207323666200428081748.
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Background: Avicennia alba Blume, is a well-known mangrove plant used in traditional medicinal practices for several human ailments. Objective: The study aimed at evaluation of antidiabetic, antioxidant, anti-inflammatory and cytotoxic activities of A. alba ethanolic leaf (AAL) and bark (AAB) extract along with phytochemical investigation. Methods: In vitro antidiabetic study was done by α-amylase, α-glucosidase enzyme inhibition assay; antioxidant study by DPPH, ABTS, superoxide, and metal chelating assays, antiinflammatory study by protein denaturation assay. The cytotoxicity study was done on TC1 murine cell line. Further, GC-MS analysis was carried out for AAL extracts. Results: AAL exhibited better antidiabetic activities with IC50 values of 1.18 and 0.87 mg/ml against α-amylase and α-glucosidase enzymes respectively. The AAL exhibited better ABTS, superoxide scavenging and metal chelating potential with IC50 values of 0.095, 0.127 and 0.444 mg/ml. However, AAB showed higher DPPH scavenging potential with IC50 value of 0.163 mg/ml. The AAL also exhibited higher protein denaturation potential with IC50 value of 0.370 mg/ml. The bark extract exhibited better cytotoxic activity as compared to leaf extracts on the TC1 murine cell line. The phytochemical study revealed higher total phenol (25.64 mg GAE/g), flavonoid (205.09 mg QE/g), and tannin content (251.17 mg GAE/g) in AAL. The GC-MS analysis revealed the presence of several compounds in AAL extract. Conclusion: The result of the present study highlights the antidiabetic, antioxidant and cytotoxic activities of mangrove plant Avicennia alba.
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Oukerrou, Moulay Ali, Mounir Tilaoui, Hassan Ait Mouse, Inass Leouifoudi, Abdeslam Jaafari, and Abdelmajid Zyad. "Chemical Composition and Cytotoxic and Antibacterial Activities of the Essential Oil of Aloysia citriodora Palau Grown in Morocco." Advances in Pharmacological Sciences 2017 (2017): 1–10. http://dx.doi.org/10.1155/2017/7801924.
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The aim of this work is to investigate the in vitro cytotoxic and antibacterial effects of the essential oils of Aloysia citriodora Palau, harvested in different regions of Morocco. The chemical profile was established using gas chromatography-mass spectrometry analysis. The cytotoxic activity against P815, MCF7, and VERO cell lines as well as the normal human peripheral blood mononuclear cells (PBMCs) was evaluated using the MTT assay. Standard, ATCC, strains of bacteria (Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa) were cultivated in Muller Hinton media. Then, agar disc diffusion, minimum inhibitory concentrations (MICs), and minimal bactericidal concentrations (MBCs) were determined using microdilution method. The essential oils obtained were predominantly composed of β-spathulenol (15.61%), Ar-curcumene (14.15%), trans-caryophyllene oxide (14.14%), and neral (10.02%). The results of the assays showed that the cytotoxic effect of the essential oil of A. citriodora was high on P815 and moderate on MCF7 and on VERO cell lines. However, no cytotoxic effect was observed on PBMCs. On the other hand, essential oils showed a significant antimicrobial activity against both Gram-negative and Gram-positive bacteria. MICs ranged between 2.84 and 8.37 mg/ml. Essential oil of A. citriodora leaves possesses significant antibacterial effect and cytotoxic activity against tumor cell lines.
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Peeri, Hadar, Nurit Shalev, Ajjampura C. Vinayaka, Rephael Nizar, Gila Kazimirsky, Dvora Namdar, Seegehalli M. Anil, Eduard Belausov, Chaya Brodie, and Hinanit Koltai. "Specific Compositions of Cannabis sativa Compounds Have Cytotoxic Activity and Inhibit Motility and Colony Formation of Human Glioblastoma Cells In Vitro." Cancers 13, no. 7 (April 5, 2021): 1720. http://dx.doi.org/10.3390/cancers13071720.
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Glioblastoma multiforme (GBM) is the most lethal subtype of glioma. Cannabis sativa is used for the treatment of various medical conditions. Around 150 phytocannabinoids have been identified in C. sativa, among them Δ-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) that trigger GBM cell death. However, the optimal combinations of cannabis molecules for anti-GBM activity are unknown. Chemical composition was determined using high-performance liquid chromatography (HPLC) and gas chromatography mass spectrometry (GC/MS). Cytotoxic activity was determined by XTT and lactate dehydrogenase (LDH) assays and apoptosis and cell cycle by fluorescence-activated cell sorting (FACS). F-actin structures were observed by confocal microscopy, gene expression by quantitative PCR, and cell migration and invasion by scratch and transwell assays, respectively. Fractions of a high-THC cannabis strain extract had significant cytotoxic activity against GBM cell lines and glioma stem cells derived from tumor specimens. A standard mix (SM) of the active fractions F4 and F5 induced apoptosis and expression of endoplasmic reticulum (ER)-stress associated-genes. F4 and F5 inhibited cell migration and invasion, altered cell cytoskeletons, and inhibited colony formation in 2 and 3-dimensional models. Combinations of cannabis compounds exert cytotoxic, anti-proliferative, and anti-migratory effects and should be examined for efficacy on GBM in pre-clinical studies and clinical trials.
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Sharma, Neha, Anket Sharma, Gaurav Bhatia, Marco Landi, Marian Brestic, Bikram Singh, Jatinder Singh, Satwinderjeet Kaur, and Renu Bhardwaj. "Isolation of Phytochemicals from Bauhinia variegata L. Bark and Their In Vitro Antioxidant and Cytotoxic Potential." Antioxidants 8, no. 10 (October 17, 2019): 492. http://dx.doi.org/10.3390/antiox8100492.
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Plants have been the basis of traditional medicine since the dawn of civilizations. Different plant parts possess various phytochemicals, playing important roles in preventing and curing diseases. Scientists, through extensive experimental studies, are playing an important part in establishing the use of phytochemicals in medicine. However, there are still a large number of medicinal plants which need to be studied for their phytochemical profile. In this study, the objective was to isolate phytochemicals from bark of Bauhinia variegata L. and to study them for their antioxidant and cytotoxic activities. The bark was extracted with methanol, followed by column chromatography and thus isolating kaempferol, stigmasterol, protocatechuic acid-methyl ester (PCA-ME) and protocatechuic acid (PCA). 2,2-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 2, 2’-diphenyl-1-picrylhydrazyl radical (DPPH) radical scavenging assays were utilized for assessment of antioxidant activity, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) dye reduction assay was used to determine cytotoxic activity against C-6 glioma rat brain, MCF-7 breast cancer, and HCT-15 colon cancer cell lines. The compounds were found to have significant antioxidant and cytotoxic activity. Since there is a considerable increase in characterizing novel chemical compounds from plant parts, the present study might be helpful for chemotaxonomic determinations, for understanding of medicinal properties as well as for the quality assessment of herbal supplements containing B. variegata bark, thus establishing its use in traditional medicine.
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Chintong, Sutasinee, Wipaporn Phatvej, Ubon Rerk-Am, Yaowapha Waiprib, and Wanwimol Klaypradit. "In Vitro Antioxidant, Antityrosinase, and Cytotoxic Activities of Astaxanthin from Shrimp Waste." Antioxidants 8, no. 5 (May 13, 2019): 128. http://dx.doi.org/10.3390/antiox8050128.
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Astaxanthin is a potent antioxidant compared with vitamins and other antioxidants. However, astaxanthin extract from shrimp processing waste has not yet been used in cosmetic products. This study aimed to explore the natural astaxanthin from shrimp shells for antioxidant and antityrosinase activities as well as potential toxicity. The antioxidant activities were performed with 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, β-carotene bleaching, and singlet oxygen quenching assays. The results revealed that astaxanthin extract demonstrated potent antioxidant activities against DPPH and ABTS radicals, and prevented the bleaching of β-carotene and quenching of singlet oxygen (EC50 17.5 ± 3.6, 7.7 ± 0.6, 15.1 ± 1.9 and 9.2 ± 0.5 μg/mL, respectively). Furthermore, the astaxanthin extract could inhibit tyrosinase activity (IC50 12.2 ± 1.5 μg/mL) and had no toxic effects on human dermal fibroblast cells. These results suggested that shrimp astaxanthin would be a promising dietary supplement for skin health applications.
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Karadeniz, Bulent, Zeynep Ulker, and Lokman Alpsoy. "Genotoxic and cytotoxic effects of storax in vitro." Toxicology and Industrial Health 29, no. 2 (December 8, 2011): 181–86. http://dx.doi.org/10.1177/0748233711428642.
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The aim of this study is to investigate the effects of the storax balsam, which is a kind of sweet gum obtained from the Liquidambar orientalis Mill trees, on cell viability, cytotoxicity and genotoxicity in human lymphocyte in vitro. We studied the genotoxic effects of the extract of storax balsam (SE) using sister chromatid exchange (SCE) test system. Also the cytotoxic and inhibitory effects on cell proliferation of SE were evaluated using lactate dehydrogenase (LDH) assay and cell proliferation (WST-1) assay. The SCE frequency was increased when the cells were treated with 1.6 and 4.0 µg/mL SE concentrations ( p < 0.05). Moreover, treatment of the cells with the same concentrations significantly depleted the cell number at 24th and 48th hours and elevated the LDH levels ( p < 0.05) at 48th hour. These results suggest that SE can be used as an alternative antibacterial and antipathogenic agent due to its cytotoxic and genotoxic effects.
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Shaikh, Muniza, Salman Siddiqui, Humaira Zafar, Uzma Naqeeb, Fakiha Subzwari, Rehan Imad, Khalid M. Khan, and Muhammad I. Choudhary. "Antiglycation Activity of Triazole Schiff’s Bases Against Fructosemediated Glycation: In Vitro and In Silico Study." Medicinal Chemistry 16, no. 4 (May 20, 2020): 575–91. http://dx.doi.org/10.2174/1573406415666190212105718.
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Background: Advanced glycation end products (AGEs) are known to be involved in the pathophysiology of diabetic complications, neurodegenerative diseases, and aging. Preventing the formation of AGEs can be helpful in the management of these diseases. Objective: Two classes of previously synthesized traizole Schiff’s bases (4H-1,2,4-triazole-4- Schiff’s bases 1-14, and 4H-1,2,4-triazole-3-Schiff’s bases 15-23) were evaluated for their in vitro antiglycation activity. Methods: In vitro fructose-mediated human serum albumin (HSA) glycation assay was employed to assess the antiglycation activity of triazole Schiff’s bases. The active compounds were subjected to cytotoxicity analysis by MTT assay on mouse fibroblast (3T3) cell line. Molecular docking and simulation studies were carried out to evaluate the interactions and stability of compounds with HSA. Anti-hyperglycemic and antioxidant activities of selected non-cytotoxic compounds were evaluated by in vitro α-glucosidase inhibition, and DPPH free radical scavenging assays, respectively. Results: Compound 1 (IC50=47.30±0.38 µM) from 4H-1,2,4-triazole-4-Schiff’s bases has exhibited antiglycation activity comparable to standard rutin (IC50=54.5±0.05 µM) along with a stable RMSD profile in MD simulation studies. Compound 1 also exhibited a potent α-glucosidase inhibitory activity, and moderate antioxidant property. Other derivatives showed a weak antiglycation activity with IC50 values between 248.1-637.7 µM. Compounds with potential antiglycation profile were found to be non-cytotoxic in a cellular assay. Conclusion: The study identifies triazole Schiff’s bases active against fructose-mediated glycation of HSA, thus indicates their potential against late diabetic complications due to production of advancedend products (AGEs).
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MILTON, Nathaniel G. N. "Amyloid-β binds catalase with high affinity and inhibits hydrogen peroxide breakdown." Biochemical Journal 344, no. 2 (November 24, 1999): 293–96. http://dx.doi.org/10.1042/bj3440293.
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Amyloid-β (Aβ) specifically bound purified catalase with high affinity and inhibited catalase breakdown of H2O2. The Aβ-induced catalase inhibition involved formation of the inactive catalase Compound II and was reversible. Catalase ↔ Aβ interactions provide rapid functional assays for the cytotoxic domain of Aβ and suggest a mechanism for some of the observed actions of Aβ plus catalase in vitro.
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Magalhães, Hemerson I. F., Paulo M. P. Ferreira, Eraldo S. Moura, Márcia R. Torres, Ana P. N. N. Alves, Otília D. L. Pessoa, Letícia V. Costa-Lotufo, Manoel O. Moraes, and Cláudia Pessoa. "In vitro and in vivo antiproliferative activity of Calotropis procera stem extracts." Anais da Academia Brasileira de Ciências 82, no. 2 (June 2010): 407–16. http://dx.doi.org/10.1590/s0001-37652010000200017.
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The cytotoxic potential of stem organic extracts from Calotropis procera (Asclepiadaceae) was firstly evaluated against cancer cell lines by MTT assay. Subsequently, samples considered cytotoxic were tested for antimitotic activity on sea urchin egg development and for in vivo antiproliferative activity in mice bearing Sarcoma 180 tumor. Among the five extracts (hexane, dichloromethane, ethyl acetate, acetone and methanol), ethyl acetate and acetone extracts displayed higher cytotoxic potential against tumor cells, with IC50 ranging from 0.8 to 4.4 μg/mL, while methanolic extract was weakly cytotoxic. Cytotoxic extracts also exhibited cell division inhibition capacity by antimitotic assay, revealing IC50 values lower than 5 μg/mL. In the in vivo antitumor assessments, ethyl acetate- and acetone-treated animals showed tumor growth inhibition ratios of 64.3 and 53.1%, respectively, with reversible toxic effects on liver and kidneys. Further studies are in progress in order to identify C. procera cytotoxic compound(s) and to understand the mechanism of action responsible for this tumor-decreasing potential.
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Fan, Chao, Hui-zi Jin, Lehao Wu, Yu Zhang, Richard D. Ye, Weidong Zhang, and Yan Zhang. "An Exploration of Traditional Chinese Medicinal Plants with Anti-Inflammatory Activities." Evidence-Based Complementary and Alternative Medicine 2017 (2017): 1–10. http://dx.doi.org/10.1155/2017/1231820.
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In a continuing effort to discover more anti-inflammatory medicinal plants in China, the anti-inflammatory activities of 101 extracts from different parts of 84 traditional medicinal plants were evaluated by a panel of in vitro and in vivo assays. Nuclear factor-kappa B (NF-κB) inhibitory effects were determined by luciferase assay in stably transfected Hela cells. Cytotoxic activities were assessed using the MTT assay. Inhibitory effects on LPS-induced nitric oxide production and proinflammatory mediators were assessed by Griess reaction and Real-Time PCR analysis, respectively. In vivo anti-inflammatory activities were examined by xylene-induced mice ear edema model. In total, 22 extracts showed promising NF-κB inhibitory effects whereas 9 of them did not affect the cell viability. The 9 hit extracts were active in at least one of the subsequently performed in vitro pharmacological test systems. The extract from Hemerocallis minor (root) was selected to perform the in vivo study because it demonstrated significant suppressive effects in all the in vitro assays. Results showed that the extract of Hemerocallis minor (Root) was able to alleviate ear edema effectively in xylene-induced mice ear edema mode. Collectively, our study provides evidence for the potential anti-inflammatory effects of the medicinal plants traditionally used in China. Further phytochemical and pharmacological studies remain to be clarified.
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Lawler, Sean, Marilin Koch, Mikolay Zdioruk, Estuardo Aguilar-Cordova, Laura Aguilar, Brian Guzik, Ghazaleh Tabatabai, Michal Nowicki, and E. Antonio Chiocca. "468 Enhancers and repressors of immunotherapy: translational perspectives on gene-mediated cytotoxic immunotherapy in glioblastoma." Journal for ImmunoTherapy of Cancer 8, Suppl 3 (November 2020): A498. http://dx.doi.org/10.1136/jitc-2020-sitc2020.0468.
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AcknowledgementsThis was supported by NCI P01CA069246 (Chiocca)ConclusionsOur data suggest that dexamethasone may decrease the efficacy of immunotherapy for glioma through impaired T cell function: this emphasizes the need in identifying alternatives to dexamethasone to prevent attenuated responses in immunotherapies. The combination of GMCI with ATRi however points to additional therapeutic benefit through enhanced cytotoxic efficacy, improved immunogenicity in vitro and increased long-term survival in vivo, making it a promising future approach for the treatment of glioblastoma.ResultsCytotoxicity assays showed that dexamethasone has a slight impact on GMCI in vitro. In T-cell-functional assays, we observed a significantly impaired tumor cell killing. Immune cell response assays revealed a reduced immune cell proliferation after co-culture with supernatant from dexamethasone or combination treated glioblastoma cells. In vivo, while treatment with GMCI alone resulted in longer median symptom-free survival (39.5d) versus no treatment (23d), the combination of GMCI and dexamethasone resulted in the significant reduction of this effect (29d vs 39.5d ; p = 0.0184).The combination of ATRi with GMCI proved to be synergistic in cytotoxicity assays. Flow cytometry revealed a significant increase in DSB-associated H2AX foci as well as an improved immune profile by downregulation of GMCI-induced PD-L1 expression. In vivo, the combination with ATRi led to an increase in long-term surviving animals (66.7%) compared to GMCI (50%) and proved to be highly significant compared to the untreated control (p=0.0022).MethodsWe investigated the effects of ATR-inhibition and dexamethasone on GMCI in vitro using cytotoxicity, flow cytometry and T-cell-killing assays in glioblastoma cell lines. The impact of dexamethasone and ATRi in vivo was assessed in an orthotopic syngeneic murine glioblastoma model. Tumor immune infiltrates were analyzed with flow cytometry.BackgroundGene-mediated cytotoxic immunotherapy (GMCI) is a local tumor immunotherapy that uses aglatimagene besadenovec (a non-replicating serotype 5 adenovirus, expressing HSV1 thymidine kinase) with the prodrug ganciclovir to induce DNA double strand breaks (DSB), leading to immunogenic tumor cell death and intratumoral immune cell invasion. Here we investigate potential repressors and enhancers of GMCI’s effectiveness. GMCI is currently in clinical trials in combination with immune checkpoint blockade in glioblastoma. Thus we set out to identify potential areas to improve this approach for future application. Dexamethasone is used in symptomatic treatment of glioma patients, although it is known to cause immune suppression. However, the influence of dexamethasone on the efficacy of GMCI has not been explored. In contrast, DNA damage response inhibitors like the ATR inhibitor (ATRi) AZD6738 might not only amend the cytotoxic but also the immunogenic profile of GMCI, rendering it an attractive combination partner.
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Akolkar, Dadasaheb, Sanket Patil, Vishakha Mhase, Pradip Devhare, Pooja Fulmali, Revati Patil, Ajay Srinivasan, et al. "INNV-26. IN VITRO CHEMO RESISTANCE PROFILES OF CIRCULATING GLIAL CELLS REPLICATE CHEMO CHARACTERISTICS OF TUMOR TISSUE." Neuro-Oncology 21, Supplement_6 (November 2019): vi135. http://dx.doi.org/10.1093/neuonc/noz175.567.
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Abstract Survival of high-grade glioma patients remains dismal due to onset of resistance to even the limited systemic treatment option currently available. Except for indirect prediction of alkylating agent Temozolomide response through MGMT promotor methylation and NTRK fusions for larotrectinib, there are no biomarkers available for drug response prediction. Cell based, in vitro chemosensitivity assays can interrogate the efficacy of an array of cytotoxic drugs. However, the unavailability of live tumor cells for such assays pose challenges in clinical practice. Repeat biopsies are neither advisable nor feasible. Access to Circulating Glial Cells (CGCs) can provide real time insight into the chemo dynamics of the tumor. In this study, we show for the first time that CGCs can be harvested from peripheral blood of glioma patients for chemo response and resistance profiles (CRR) of cytotoxic drugs. CGCs were harvested from 15 ml of peripheral blood from high grade GBM patients (n=9) out of whom cells derived from surgically excised tumor tissue were also available for comparison in 2 patients. CellWizard™ process was adopted for enrichment of CGCs which is based upon epigenetically active media with paradoxical chemo-toxicity that selectively induces lethality in normal cells. This paradoxical cytotoxicity of the medium leads to selective elimination of most leukocytes thus facilitating a label free negative enrichment of CGCs. In vitro chemo sensitivity assay performed on live CGCs and cell death events were determined to evaluate response to different class of chemotherapy drugs. Evaluation of drug response showed very high concordance between tumor derived cells and CGCs in both patients where live tissue was available. In 7 patients where CGCs alone could be evaluated, the response showed replication between in vitro profile compared to treatment antecedents in 5 patients. 2 patients were treatment naïve and the response reflected high sensitivity to Temozolomide.