scholarly journals Evaluating the in vitro Cytotoxicity of Thymus vulgaris Essential Oil on MCF-7 and HeLa Cancer Cell Lines

2021 ◽  
pp. 2862-2871
Author(s):  
Alsafa N. Khalaf ◽  
Ibrahim J. Abed
Keyword(s):  
Essential Oil ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Thymus Vulgaris ◽  
Hela Cell Line ◽  
Significant Difference ◽  
Mcf 7

     Thyme essential oil (TEO) was extracted from dried leaves of Thymus vulgaris. The air-dried aerial parts of the plant produced 1.0% yield of TEO. The detection of this essential oil’s compounds was performed by GC-MASS. The cytotoxic activity of TEO was evaluated against two human cancer cell lines, namely HeLa (human epithelial cervical cancer) and MCF-7 (human breast carcinoma). Cells grown in 96 multi-well plates were treated with six concentrations of EO (6.25, 12.5, 25, 50, 100, 200 ppm) and incubated at 37 °C for 72 hrs. Cancer cell lines elicited various degrees of sensitivity to the cytotoxic effect of essential oil. The TEO exhibited significant differences (p≤ 0.01) between the effects of all concentrations against these two human cell lines. The results showed the highest toxicity of TEO on HeLa cell line (78.67%) and MCF-7 cell line (83.60%) at 200 ppm concentration. Also the values of half-maximal inhibitory concentration (IC50) of TEO against HeLa and MCF-7 cell lines were 34.63 and 27.66 ppm, respectively. Cells treated with the IC50 of TEO showed a significant difference (p ≤ 0.01) in p53 fold expression between HeLa cell line (4.33±0.41 folds) and MCF-7 cell line (5.10±0.32 folds). In general, a dose-dependent decrease the survival of the two cell lines was observed. In addition, MCF-7 cell line revealed higher sensitivity against TEO than HeLa cell line.

Effect of the Method of Preparation on the Composition and Cytotoxic Activity of the Essential Oil of Pituranthos tortuosus

2011 ◽  
Vol 66 (3-4) ◽  
pp. 143-148 ◽  
Author(s):  
Hossam M. Abdallah ◽  
Shahira M. Ezzat
Keyword(s):  
Breast Cancer ◽  
Essential Oil ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Human Cancer ◽  
Cancer Cell Line ◽  
Cancer Cell Lines ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines

The aerial parts of Pituranthos tortuosus (Desf.) Benth and Hook (Apiaceae), growing wild in Egypt, yielded 0.8%, 0.6%, and 1.5% (v/w) of essential oil when prepared by hydrodistillation (HD), simultaneous hydrodistillation-solvent (n-pentane) extraction (Lickens- Nickerson, DE), and conventional volatile solvent extraction (preparation of the “absolute”, SE), respectively. GC-MS analysis showed that the major components in the HD sample were β-myrcene (18.81%), sabinene (18.49%), trans-iso-elemicin (12.90%), and terpinen- 4-ol (8.09%); those predominent in the DE sample were terpinen-4-ol (29.65%), sabinene (7.38%), γ-terpinene (7.27%), and β-myrcene (5.53%); while the prominent ones in the SE sample were terpinen-4-ol (15.40%), dill apiol (7.90%), and allo-ocimene (4E,6Z) (6.00%). The oil prepared in each case was tested for its cytotoxic activity on three human cancer cell lines, i.e. liver cancer cell line (HEPG2), colon cancer cell line (HCT116), and breast cancer cell line (MCF7). The DE sample showed the most potent activity against the three human cancer cell lines (with IC50 values of 1.67, 1.34, and 3.38 μg/ml against the liver, colon, and breast cancer cell lines, respectively). Terpinen-4-ol, sabinene, γ-terpinene, and β-myrcene were isolated from the DE sample and subjected to a similar evaluation of cytotoxic potency; signifi cant activity was observed

scholarly journals The Effect of Okra (Abelmoschus esculentus (L.) Moench) Seed Extract on Human Cancer Cell Lines Delivered in Its Native Form and Loaded in Polymeric Micelles

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Watcharaphong Chaemsawang ◽  
Weerapong Prasongchean ◽  
Konstantinos I. Papadopoulos ◽  
Garnpimol Ritthidej ◽  
Suchada Sukrong ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Polymeric Micelles ◽  
Cancer Cell Lines ◽  
Seed Extract ◽  
Abelmoschus Esculentus ◽  
Native Form ◽  
Flavonoid Compounds ◽  
Mcf 7

Cancer is a noncommunicable disease with a high worldwide incidence and mortality rate. The National Cancer Institute of Thailand reports increasing cumulative incidence of breast, colorectal, liver, lung, and cervical cancers, accounting for more than 60% of all cancers in the kingdom. In this current work, we attempt to elucidate the phytochemical composition of the okra (Abelmoschus esculentus (L.) Moench) seed extract (OSE) and study its anticancer activity, delivered in its native form as well as in the form of polymeric micelles with enhanced solubility, in three carcinoma cell lines (MCF-7, HeLa, and HepG2). The presence of flavonoid compounds in the OSE was successfully confirmed, and direct delivery had the highest cytotoxic effect on the breast cancer cell line (MCF-7), followed by the hepatocellular carcinoma (HepG2) and cervical carcinoma (HeLa) cell lines in that order, while its delivery in polymeric micelles further increased this effect only in the HepG2 cell line. The OSE’s observed cytotoxic effects on cancer cell lines demonstrated a dose and time-dependent cell proliferation and migration inhibition plausibly due to VEGF production inhibition, leading to apoptosis and cell death, conceivably due to the four flavonoid compounds noted in the current study, one of which was isoquercitrin. However, in view of the latter compound’s isolated effects being inferior to those observed by the OSE, we hypothesize that either isoquercitrin requires the biological synergy of any one or all of the observed flavonoids or any of the three in isolation or all in concert are responsible. Further studies are required to elucidate the nature of the three unknown compounds. Furthermore, as we encountered significant problems in dissolving the okra seed extract and creating the polymeric micelles, further studies are needed to devise a clinically beneficial delivery and targeting system.

PRELIMINARY CYTOTOXIC EVALUATION OF Andrographis paniculata IN BREAST CANCER CELL LINES

2016 ◽  
Vol 2 (1) ◽  
pp. 34
Author(s):  
Tarwadi . ◽  
Churiyah . ◽  
Olivia Bunga Pongtuluran ◽  
Fifit Juniarti ◽  
Fery Azis Wijaya
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Cancer Cell Lines ◽  
Andrographis Paniculata ◽  
Normal Fibroblast ◽  
Breast Cancer Cell Lines ◽  
Mcf 7

Sambiloto (Andrographis paniculata) banyak digunakan untuk mengobati berbagai penyakit di Indonesia dan negara-negara Asia lainnya. Dalam studi ini, ekstrak metanol dan etanol sambiloto yang diperoleh dari B2PTO Tawangmangu telah diuji terhadap sel lini kanker payudara T47D dan MCF-7 dan sel lini normal fibroblast HFL-1 menggunakan reaksi enzimatik 3-(4,5-dimethylthiazoyl-2-yl) 2,5-diphenyltetrazoliumbromide (MTT). Uji in vitro terhadap sel lini normal fibroblast HFL-1 menunjukkan bahwa 50 ppm ekstrak metanol sambiloto tidak menghambat pertumbuhan sel. Tetapi, ekstrak metanol dan etanolnya menghasilkan IC50 yang relatif rendah pada sel lini kanker payudara, yaitu 111 ppm dan 122 ppm pada sel lini MCF-7 dan 70 ppm dan 197 ppm pada sel lini T47D. Selain itu, campuran ekstrak sambiloto yang mengandung 25% ekstrak Thyponium divaricatum dan Anredera cordifolia memberikan daya hambat pertumbuhan pada sel kanker payudara MCF-7 yang lebih besar, dengan nilai IC50 masing-masing adalah 68 ppm dan 34 ppm. Kesimpulannya, total ekstrak metanol atau etanol sambiloto yang diperoleh dari Tawangmangu memiliki potensi sebagai sumber senyawa anti-kanker serta perlu kajian lebih lanjut.Kata kunci: Ekstrak Andrographis paniculata, MTT, sel lini normal, sel lini kanker, aktivitas anti kanker ABSTRACTSambiloto (Andrographis paniculata) is widely used as medicine to treat various diseases in Indonesia and other Asian countries. In this study, methanolic and ethanolic extracts of sambiloto collected from B2PTO Tawangmangu have been tested againts breast cancer cell lines of T47D and MCF-7 and normal fibroblast cell line of HFL-1 using enzymatic reaction of 3-(4,5-dimethylthiazoyl-2-yl) 2,5-diphenyltetrazoliumbromide (MTT). In vitro assay performed on normal fibroblast of HFL-1 cell line showed that 50 ppm of methanolic extract of sambiloto did not inhibit cell growth. However, methanolic and ethanolic extracts of sambiloto gave relatively low of IC50 on breast cancer cell lines which were 111 ppm and 122 ppm on the MCF-7 cell lines and 70 ppm and 197 ppm on the T47D cell lines, respectively. In addition, the mixture of sambiloto extract containing 25% of Thyponium divaricatum and Anredera cordifolia extracts confered greater growth inhibition on breast cancer cell line of MCF-7, where IC50 values were 68 ppm and 34 ppm, respectively. In conclusion, the total methanolic or ethanolic extract of sambiloto collected from Tawangmangu has potency as a source of anti-cancer compounds and needs further study.Key words: Andrographis paniculata extract, MTT, normal cell line, cancer cell lines, anti-cancer activity

scholarly journals Synthesis and Biological Activity Studies of Methyl-5-(Hydroxymethyl)-2-Furan Carboxylate and Derivatives

2019 ◽  
Vol 35 (3) ◽  
pp. 080-1085 ◽  
Author(s):  
Weerachai Phutdhawong ◽  
Siwaporn Inpang ◽  
Thongchai Taechowisan ◽  
Waya S. Phutdhawong
Keyword(s):  
Biological Activity ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Furfuryl Alcohol ◽  
Methyl Acetate ◽  
Biological Activities ◽  
Hela Cell Line ◽  
Amine Derivative

Methyl-5-(hydroxymethyl)-2-furan carboxylate and derivatives were prepared from furfuryl alcohol and their biological activities were studied for cytotoxicity against cancer cell lines HeLa, HepG2 and Vero, and Gram (+) and Gram (-) bacteria. The amine derivative, (5-(((2-(1H-indol-3-yl)ethyl)amino)methyl) furan-2-yl)methyl acetate, was found to have the most potent biological activity with IC50 62.37 µg/mL against the HeLa cell line and MIC 250 µg/mL against the photogenic bacteria.

scholarly journals Antiproliferative, antimicrobial, and antioxidant activity of Lavandula angustifolia Mill. essential oil

2017 ◽  
Vol 7 (1) ◽  
pp. 35-43 ◽  
Author(s):  
Haris Niksic ◽  
Elvira Kovac-Besovic ◽  
Elma Omeragic ◽  
Samija Muratovic ◽  
Jasna Kusturica ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Essential Oil ◽  
Cell Lines ◽  
Cancer Cell ◽  
Antiproliferative Activity ◽  
Human Cancer ◽  
Cancer Cell Lines ◽  
Gas Chromatography Mass Spectrometry ◽  
Lavandula Angustifolia ◽  
Mcf 7

Introduction: We studied the chemical composition and antimicrobial, antioxidant, and antiproliferative activities of essential oils from flowers of Lavandula angustifolia grown in Southern Bosnia and Herzegovina. Methods: The chemical profile of essential oil was evaluated by means of gas chromatography-mass spectrometry. Antimicrobial activity was tested against six bacterial strains. The antioxidant activity by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) test and the antiproliferative activity against three human cancer cell lines, MCF-7, NCI-H460, and MOLT-4, were investigated using 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide tests. Results: In L. angustifolia essential oil, monoterpene alcohols were the most represented class of volatiles (51.8%), including linalool, lavandulol, and terpinen-4-ol, α-terpineol as the major components, followed by monoterpene esters (22.6%). The most important antibacterial activity of essential oil was expressed on Gram-negative strains. Investigated essential oil was able to reduce DPPH radicals into the neutral DPPH-H form (inhibitory concentration 50% [IC50] = 0.421 mg/ml), and this activity was dose dependent. The essential oil showed significant antiproliferative activity against three cancer cell lines, MOLT-4, MCF-7, and NCI-H460 cells, with IC50 values of 17, 94, and 97 µg/ml, respectively. The result of the antiproliferative assay indicates that MOLT-4 cell line was the most sensitive to investigated essential oil. Conclusion: The results revealed that L. angustifolia essential oil may be important growth inhibitor against the microbes studied. It also possesses significant antioxidant activity and demonstrated excellent antiproliferative activity against MOLT-4 cells.

scholarly journals Cytotoxicological Investigation of the Essential Oil and the Extracts of Cotula cinerea and Salvia verbenaca from Morocco

2018 ◽  
Vol 2018 ◽  
pp. 1-5 ◽  
Author(s):  
Fatima-Ezzahrae Guaouguaou ◽  
Mohamed Alien Ahl Bebaha ◽  
Khalid Taghzouti ◽  
Abdelhakim Bouyahya ◽  
Youssef Bakri ◽  
...  
Keyword(s):  
Essential Oil ◽  
Ethyl Acetate ◽  
Vero Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Antitumor Agents ◽  
Cancer Cell Lines ◽  
Hexane Extract ◽  
Cotula Cinerea

The objective of this work was to investigate the cytotoxicological effect of the extracts (hexane, ethyl acetate, and n-butanol) of Cotula cinerea and Salvia verbenaca in addition to the essential oil of Cotula cinerea. These plants are widely used in the Moroccan traditional folk medicine. The cytotoxic effect was explored against two cancer cell lines, Vero and RD, using the colorimetric MTT assay. The obtained results showed that the cytotoxicity differed according to the used extract with an efficient effect of Cotula cinerea extracts compared to Salvia verbenaca. A potent cytotoxicity was thus observed for the Cotula cinerea hexane extract which inhibited the growth of RD cell line at the lowest IC50 value (57.21±3.43 µg/mL). This was followed by the ethyl acetate extract and the essential oil with moderate effects against RD cell line and showed IC50 values of 187.52±6.27 µg/mL and 173.05±4.46 µg/mL, respectively. On the other hand, different results were obtained and Cotula cinerea essential oil was the most cytotoxic with the lowest IC50 value (72.72±2.18 µg/mL) against Vero cell line. In the same conditions, higher concentrations were needed in the case of Salvia verbenaca extracts. The results of this study showed thus that Cotula cinerea essential oil and hexane extract showed significant cytotoxic effects against RD and Vero cell lines, respectively, and could be considered as novel source of antitumor agents. This study is expected to be beneficial for clinical and traditional applications for Cotula cinerea as a remedy against cancer and opens new perspectives for further investigations on other types of cancer cell lines.

scholarly journals Diorganotin Complexes of a Thiosemicarbazone, Synthesis: Properties, X-Ray Crystal Structure, and Antiproliferative Activity of Diorganotin Complexes

2010 ◽  
Vol 2010 ◽  
pp. 1-9 ◽  
Author(s):  
Joanna Wiecek ◽  
Dimitra Kovala-Demertzi ◽  
Zbigniew Ciunik ◽  
Maria Zervou ◽  
Mavroudis A. Demertzis
Keyword(s):  
Hydrogen Bonds ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Cancer Cell Lines ◽  
Bladder Cancer Cell Line ◽  
Intermolecular Hydrogen Bonds ◽  
X Ray ◽  
Mcf 7

The synthesis and spectral characterization of novel diorganotin complexes with 3-hydroxypyridine-2-carbaldehyde thiosemicarbazone,(1), [(L)](2), [(L)](3), and [(L)](4)are reported. The single-crystal X-ray structure of complex [(L)(DMSO)](5)shows that the ligand is doubly deprotonated and is coordinated as tridentate ligand. The six coordination number is completed by two carbon atoms of phenyl groups. There are two similar monomers5a(Sn1) and5b(Sn51) in the asymmetric unit. The monomers5aand5bare linked through intermolecular hydrogen bonds of N–H–O and C–H–S type. , intermolecular interactions, intra- and intermolecular hydrogen bonds stabilize this structure and leads to aggregation and a supramolecular assembly. The IR and NMR (, and ) spectroscopic data of the complexes are reported. The in vitro cytotoxic activity has been evaluated against the cells of three human cancer cell lines: MCF-7 (human breast cancer cell line), T-24 (bladder cancer cell line), A-549 (nonsmall cell lung carcinoma) and a mouse L-929 (a fibroblast-like cell line cloned from strain L). Compounds1,3, and4were found active against all four cell lines. Selectivity was observed for complexes3and4which were found especially active against MCF-7 and T-24 cancer cell lines.

scholarly journals Ethanolic extract of Nymphaea lotus L. (Nymphaeaceae) leaves exhibits in vitro antioxidant, in vivo anti-inflammatory and cytotoxic activities on Jurkat and MCF-7 cancer cell lines

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Benoit Banga N’guessan ◽  
Adwoa Dede Asiamah ◽  
Nana Kwame Arthur ◽  
Samuel Frimpong-Manso ◽  
Patrick Amoateng ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Ethanolic Extract ◽  
Cancer Cell Lines ◽  
Paw Oedema ◽  
Anti Inflammatory ◽  
Chang Liver ◽  
Mcf 7

Abstract Background Nymphaea lotus L. (N. lotus) is an aquatic plant with anecdotal reports suggesting its use in the traditional management of cancer. However, there is a paucity of data on the antioxidant, anti-inflammatory and cytotoxic properties of N. lotus in relation to its phytochemical and elemental contents. This study aimed at determining the antioxidant, anti-inflammatory and cytotoxic properties of the hydro-ethanolic extract of N. lotus leaves (NLE), and its phenolic, flavonoid and elemental constituents. Methods The antioxidant property of NLE was determined using total phenolic and flavonoid, DPPH radical scavenging, lipid peroxidation and reducing power assays. The anti-inflammatory activity of NLE (100–250-500 mg/kg), diclofenac and hydrocortisone (positive controls) were determined by paw oedema and skin prick tests in Sprague Dawley rats. Also, the erythrocyte sedimentation rate (ESR) was determined by Westergren method. The macro/micro-elements content was determined by the XRF method. The cytotoxic property of NLE was determined by the MTT assay, on two cancer cell lines (MCF-7 and Jurkat) and compared to a normal cell line (Chang liver). Inhibitory concentrations were determined as IC50 values (±SEM). Results The extract had appreciable levels of phenolic and flavonoids compounds and was two-fold more potent in scavenging DPPH radicals than Butylated hydroxytoluene (BHT). However, NLE was three- and six-fold less potent than ascorbic acid and BHT, respectively, in reducing Fe3+ to Fe2+. The extract was six-fold more potent than gallic acid in inhibiting lipid peroxidation. The extract caused a dose-dependent decrease in rat paw oedema sizes, comparable to diclofenac, and a significant decrease in wheel diameters and ESR. The elemental analysis revealed relevant concentrations of Mg2+, P2+, S2+, K2+, Mn+, Fe+, Cu+, Zn+ and Cd+. The extract exhibited cytotoxic activity on both MCF-7 (IC50 = 155.00 μg/ml) and Jurkat (IC50 = 87.29 μg/ml), with higher selectivity for Jurkat cell line. Interestingly, the extract showed low cytotoxicity to the normal Chang liver cell line (IC50 = 204.20 μg/ml). Conclusion N. lotus leaves extract exhibited high antioxidant, anti-inflammatory and cancer-cell-specific cytotoxic properties. These aforementioned activities could be attributed to its phenolic, flavonoid and elemental constituents. Graphical abstract

scholarly journals Regio- and Stereoselective Synthesis of a New Series of Spirooxindole Pyrrolidine Grafted Thiochromene Scaffolds as Potential Anticancer Agents

Symmetry ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1426
Author(s):  
Assem Barakat ◽  
Mohammad Islam ◽  
M. Ali ◽  
Abdullah Al-Majid ◽  
Saeed Alshahrani ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Anticancer Agents ◽  
Stereoselective Synthesis ◽  
Cancer Cell Lines ◽  
Azomethine Ylides ◽  
Breast Cancer Cell Lines ◽  
Active Member ◽  
Mcf 7

A series of new spiro-heterocycles engrafted spirooxindole/pyrrolidine/thiochromene scaffolds was synthesized by the three-component 1,3-dipolar cycloaddition reactions in a fully controlled regio- and stereo-selective fashion. Condensation of several substituted isatin derivatives with L-proline generated the azomethine ylides which subsequently reacted with chalcones based thiochromene scaffold, and finally afforded the target spiro-compounds. This simple protocol furnished a structurally complex, biologically relevant spiro-heterocycles in good yields through a one-pot process. All synthesized chalcone-based thiochromene, along with the spirooxindole/pyrrolidine/thiochromene scaffolds, were tested for their anticancer activity against four cancer cell lines (PC3, HeLa, MCF-7, and MDA-MB231). Toxicity of these compounds was also evaluated against human fibroblast BJ cell line, and they appeared to be not cytotoxic. For the prostate cancer (PC3) cell line, the most active hybrid, among synthesized series, was compound (7f,IC50 = 8.7 ± 0.7 µM). The most potent spirooxindole/pyrrolidine/thiochromene hybrid against cervical (HeLa) cancer cells was compound (7k, IC50 = 8.4 ± 0.5 µM) having chlorine and p-trifluoromethyl substituents attached to phenyl rings. Finally, against the MCF-7 and MDA-MB231 breast cancer cell lines, compound (7d) was the most active member of this series (IC50 = 7.36 ± 0.37, and 9.44 ± 0.32 µM, respectively).

scholarly journals Synthesis and Biological Evaluation of Phaeosphaeride A Derivatives as Antitumor Agents

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 3043 ◽  
Author(s):  
Victoria Abzianidze ◽  
Petr Beltyukov ◽  
Sofya Zakharenkova ◽  
Natalia Moiseeva ◽  
Jennifer Mejia ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Antitumor Agents ◽  
Cancer Cell Lines ◽  
Biological Evaluation ◽  
Tumor Cell Lines ◽  
Hct 116 ◽  
Mcf 7

New derivatives of phaeosphaeride A (PPA) were synthesized and characterized. Anti-tumor activity studies were carried out on the HCT-116, PC3, MCF-7, A549, К562, NCI-Н929, Jurkat, THP-1, RPMI8228 tumor cell lines, and on the HEF cell line. All of the compounds synthesized were found to have better efficacy than PPA towards the tumor cell lines mentioned. Compound 6 was potent against six cancer cell lines, HCT-116, PC-3, K562, NCI-H929, Jurkat, and RPMI8226, showing a 47, 13.5, 16, 4, 1.5, and 7-fold increase in anticancer activity comparative to those of etoposide, respectively. Compound 1 possessed selectivity toward the NCI-H929 cell line (IC50 = 1.35 ± 0.69 μM), while product 7 was selective against three cancer cell lines, HCT-116, MCF-7, and NCI-H929, each having IC50 values of 1.65 μM, 1.80 μM and 2.00 μM, respectively.

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