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Glenski, T. Shah, N. AlJasser, R. Golla, A. Bjurlin, M. Viscusi, W. Park, S. Oldham, M. Pulvers, K. Bekki, K. Dewhirst, T. Bricknell, R. Metzen, D. Yoong, S. Jordt, S. Kang, H. Lin, S. Park, H. Shah, S. Miglio, F. Wright, C. Hair, E. Zhou, Y. Luo, J. Huh, J. Leas, E. Fucito, L. Gaiha, S. Jutla, S. Boccio, C. Sanchez, R.

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Molino, A. Selya, A. Henderson, B. Swann, A. Garrett, P. Wu, N. Lanza, H. Sharma, E. Vlahos, R. Vena, A. Vargas-Rivera, M. Lempert, L. Churchill, V. Maloney, S. Erythropel, H. Sussman, S.

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Cao, Y. Mallock, N. Lee, Y. Hammond, D. Kaplan, B. Loffredo, L. Pacek, L. Mostafa, M. Airagnes, G. Liautaud, M. Buckner, J.

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Morel Espinosa, M. Ali, F. Smith, D. Doxbeck, C. Helen, G. Cox, S. Pacula, R. Hall, W. Wong, E. Levy, D. Klawinski, D. Eversole, A. Camara-Medeiros, A. Seidenberg, A. Kwon, E. Fu, R. Fahey, M. Kennedy, P. Rudasingwa, G. Coleman, S. Pincus, J.

Williams, M. Amalia, B. Watson, C. Sun, Y. Borgini, A. Martheswaran, T. Kreslake, J. Reddy, K. Cuomo, R. Pappas, R. Cobb, C. Escobar, Y. Majmundar, A. Li, L. Massey, M. Durazo, A. Yan, B. Moustafa, A. Hassoun, A. Stroup, A. Schwarzmeier, L. Alqahtani, M. Xu, G. Ma, C. Rouabhia, M. Lee, S. Rycroft, N. Villanti, A. Sokolovsky, A. Shin, S. Huang, J. Knapp, A. Ronchetti, J. Wu, J. Rogers, S. Algiers, O. Schillo, B. Sahota, A.

Gwon, S. Ikegwuonu, T. Holliday, R. Tattan-Birch, H. Prochaska, J. Olonoff, M. Hamad, S. Felicione, N. Alpert, J. Wackowski, O. Czekala, L. Sarles, S. Wang, T. Shi, R. Kim, D. Verma S. Allami, R. Khadka, S. Miller, C. Shamout, M. Culbreth, R. Yang, B. Xian, S. Liu, B. Powers, J. Keith, R. Syamlal, G. Godby, S. Gugala, E. Driller, G. Yan, R. Cohn, A. Kozlovich, S. Samuolis, J. Bracken-Clarke, D. McClelland, M.

Kwak, S. Tercyak, K. Kosterman, R. Cooper, S. Keller-Hamilton, B. Phillips-Waller, A. Kim, Y. Siripongvutikorn, Y. Strombotne, K. Shahab, L. Lozano, A. Abadi, M. De Martin, S. Choi, K. Baker, A. El-Merhie, N. Ishii, K. El-Mahdy, M. Snell, M. Anic, G.

Fried, N. Lucero, A. Imura, Y. Stevens, E. Duan, Z. Tam, J. Szparaga, M. Hershberger, A. Rigotti, N. Jin, L. Zhu, S. Bares, C. Hernandez, M. Sussman, R. Perez, M. Bold, K. Tai, H. Chen, H. Hasan, K. Bandara, N. Patten, S. Schilling, L. Khan, A. L JL. PT JL. BASIA jl. RANDY NUR M. KONCI POLTEKES JL. BIDA Hi. AMIN M. P JL. IPI NO. PD JL. MG JL. ST JL. M JL. SI JL. FT P JL. AG JL. MA JL. KEP JL. KES JL. MD JL. LB 48 M. ALWI 87 M. ARI M. NUR JL. JAYA M. HUT JL. B P JL. T JL.

PD KEL. FAR JL. PD JL.. MK JL. PD jl. Mamboro,27 Februari Dokter Keluarga. Tenrisa'na R Nip. Alergi x x. Samlia Kurase 55th Hipertensi x x. A x x 19 Fera 19th Sus. Tanggal 21 April 34 Jodjo djoni 70th R. A x x 35 Limin jaleha 59th konjunctivitis x rjk 36 Novitasari 35th Sus. Alergi x x 38 Rese Ilyas 53th Hipertensi x x 39 M.

Hymenocrater longiflorus is a dense bush with a strong characteristic aroma and green leaves. Hymenocrater longiflorus is utilized as a medicinal herb in local and traditional medicine especially in Kurdistan. The aerial parts of the herb, in crude or baked form, are utilized in local folk medicine as an anti-inflammatory, sedative, and anti-allergy treatment for skin diseases and insect bites.

Recently, the antibacterial, antifungal, and antioxidant activities of H. However, no report exists regarding the cytotoxic and antitumor properties of H. For this reason, the present study was designed to investigate the in vitro anti-tumor activities of the methanolic extract of H.

The chemical composition and antioxidant activity of the extract were studied as well. Methanol, ethanol, and acetic acid were purchased from Carlo Erba Cornaredo, Italy. The anti-p53 monoclonal antibody Clone sc , anti-p21 polyclonal antibody produced in rabbit Clone sc , and anti-actinin monoclonal antibody Clone sc were purchased from Santa Cruz Biotechnology Dallas, TX, USA. The mouse anti-parp monoclonal antibody Clone sc and goat anti-actin monoclonal antibody Clone sc were purchased from Santa Cruz Biotechnology.

Hymenocrater longiflorus Benth Lamiaceae was collected from the Hawraman Mountain m above sea level above the Ballkha village in the Kurdistan Province of Iraq in May The plant material was identified by Dr. The fresh aerial parts of the plant were washed thoroughly with tap water at room temperature to remove dirt prior to drying. The washed aerial parts were dried in the shade at room temperature for 7 d and then crushed into a fine powder. After filtering through Whatman filter paper no.

For in vitro studies, the H. The H. The ion source was a desorption electrospray ionization Omni spray Prosolia, Indianapolis, IN, USA , which was used in nanoelectrospray mode for negative and positive ions. Samples were directly injected into the HPLC column, which was directly coupled to the ion source spray capillary by a liquid junction. The available reference standards were used apigenin, caffeic acid and acacetin for identification by direct comparison of the retention times and mass spectra acquired in the same analytical conditions.

For other peaks, the measured accurate mass or the most probable chemical formula that can be calculated from it was compared with reference compounds available online at different chemical database websites www. The free radical scavenging activities of the plant extract were measured with the DPPH assay [13]. The DPPH 2,2-diphenylpicrylhydrazyl radical has a deep violet color because of its unpaired electron; the radical scavenging capability can be determined spectrophotometrically by the absorbance loss at nm, when the pale yellow non-radical form is produced.

Equal volumes 0. The absorbance of the control DPPH solution was also read. The percentage of DPPH decoloration of the sample was calculated using the following formula:. After each incubation period, cells were counted, and the viability was assessed using a cell counter [14]. The MTT assay was used for confirmation.

Subsequently, MTT dissolved in phosphate buffered saline PBS at 5 mg mL —1 was added to each well and incubated for 60 min in the dark. After 5 min incubation, absorbance was measured at nm in a Biotex microplate reader.

The intensity of color produced is directly proportional to the number of viable cells. Each experiment was performed in triplicate [15]. Analysis of the cell cycle was performed according to the method of Erba et al. A negative control was run as well. The cells were suspended in PBS, mixed, and incubated for 30 min in an ice bath. The data were analyzed with CellQuest and ModFit 3.

Subsequently, the medium was discarded by aspiration. The cells were washed with filtered PBS and stained with sulforhadamineDAPI 4,6-diamidinephenylindole dihydrochloride solution for 30—60 min. After staining, the slides were rinsed with distilled water, air-dried, and covered with a cover slip.

Nuclei were observed with a fluorescence microscope by using a nm filter [18]. After treatment, the medium was removed, and the cells were washed with PBS two to three times. Subsequently, the cells were re-hydrated and permeabilized with 0. They were subsequently inspected under a fluorescence microscope TE Eclipse , Nikon. Images were acquired using Vico software. For immunoblotting, U2OS cells were grown under the same conditions described above and treated with H. The nitrocellulose membranes were immediately stained in 0.

This step was followed by 15 min washes with TBS-T prior to 1 and 0. Data were obtained by tabulation in the statistical program GraphPad Prism version 5. The most abundant compounds were cirsimaritin, rosmarinic acid, apigeninO-glucoside, genistein, and apigenin, followed by acacetin, carnosic acid, caffeic acid, ferulic acid, and isorhamnetin.

The free radical scavenging activity of the H. Such potent antiradical and antioxidant activity appears to be correlated with the total amount of polyphenolic compounds. Ahmadi et al. Chemical compounds identified in the H. DPPH free radical scavenging activity on different concentrations of the H.

Negative control absorbance, DPPH 0. Flavonoids are considered the most abundant and most effective antioxidant compounds in plants. Present findings suggest that apigenin and related flavonoids are potentially useful as antioxidant and may have therapeutic applications in various diseases.

Many natural phenols have been reported to scavenge reactive oxygen species ROS , which are implicated in DNA damage, cancer, and accelerated cell aging [20, 21], while others can induce differentiation of numerous cell types [22] and inhibit glutathione reductase, cytochrome P, and topoisomerase I-catalyzed DNA relegation [23].

FACS analysis revealed that the H. Many flavonoids have been shown to target the cell cycle by affecting cycle checkpoint pathways as well as the DNA damage response DDR pathway [24].

Mean of cell number of U2OS cells treated with different concentrations of the H. Results are from one experiment that is representative of three similar experiments. NT, Non treated cells. Cells were harvested 3 or 6 h after addition of the extract and assayed for DNA content by propidium iodide PI staining; flow cytometry was then performed for cell cycle distribution. Percentage of cell distribution data for each treatment is indicated. As seen in Figure 4 , drastic changes in cellular morphology, such as a notable shrinkage of the cells, as well as aberrant nuclei were observed in response to the H.

The observed antiproliferative effects Table 3 ; Figure 2 prompted us to investigate possible DNA damage in treated cells. The region flanking a DSB appears as a spot or focus when microscopically examined after antibody staining.