Journal of Natural Products
Article
2
h after irradiation; isolated plant compounds were added at the same
time.
Micronucleus Analysis. The blood lymphocytes were set up in
Atriplexin III (3): yellow powder; [α]22 −22 (c 1 MeOH); UV λmax,
D
(MeOH) 240, 270, 346 nm, 270, 394 (NaOMe), 246, 270, 286, 384
(AlCl ); 246, 270, 286, 384 (AlCl + HCl), 260, 280, 348 (NaOAc),
3
3
RPMI-1640 medium, supplemented with calf serum and phytohemag6-
glutinin (Invitrogen−Gibco-BRL, Vienna, Austria). A level of 2 × 10
of human lymphocytes was chosen because it corresponds to 0.5 mL
of human blood. The concentration of each compound (six isolated
phenolic compounds and the positive and negative controls) added to
the cell cultures was adjusted to correspond to the total blood amount
of an average adult, estimated as approximately 5 L. The isolated
compounds at a concentration range of 0.8−7.4 μM were added to the
irradiated samples and were used as usual therapeutic doses. The
concentration of amifostine of 4.7 μM was determined to be optimal
in the in vitro experiment; higher concentrations were assumed to be
toxic.
260, 280, 348 (NaOAc + H BO ); IR ν 3428, 2936, 1680, 1621,
3 3 max
1565, 1514, 1478, 1392, 1346, 1256, 1216, 1130, 1101, 1072, 1020,
−
1
1
13
828 cm ; H NMR and C NMR data, see Table 1; HRESIMS m/z
−
651.1606 [M − H] (calcd for C H O −H 651.1567).
29
32 17
ASSOCIATED CONTENT
Supporting Information
■
*
S
Spectroscopic data of the isolated compounds (PDF)
One cell culture served as a control, not containing compounds
isolated from A. littoralis. The cell culture with added synthetic
radioprotectoramifostine WR-2721 (98%, S2[3-aminopropylami-
no]-ethylphosphothioic acid; obtained from Marlingen-Biosciences,
USA) at a concentration of 4.7 μMserved as the positive control for
AUTHOR INFORMATION
Corresponding Author
■
*
3
5
comparison with isolated plant phenolic compounds, as reported.
Because the radioprotective mechanisms of amifostine, including
interactions and effects on alkaline phosphatase activity, have been
reported, especially when radioprotectivity of amifostine was analyzed
in vivo and by the comet test, they were of no interest here and thus
were not analyzed. One cell culture included mitomycin C (MMC)
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
(
3.0 μM, in phosphate buffer), an alkylating agent that served as the
This study was financially supported by the Serbian Ministry of
Education and Science, Project No. 172053. The authors are
grateful for support of EU Commission project AREA, No.
316004. The authors thank N. Brierley, Professor at Brierley &
Co. Language Centre, Belgrade, as well as the ACS English
Editing Service for review and correction of English style and
grammar.
negative control.
All cultures were incubated at 37 °C. Treatment with flavonoids
lasted for 19 h. All cultures (including amifostine WR-2721 and
MMC) were rinsed with pure medium, transferred to fresh RPMI
1
640 medium, and incubated for a duration of 72 h.
DNA damage was measured using the cytokinesis-block micro-
nucleus assay, enabling determination of MN in cells that had
completed nuclear division and thus were not influenced by variations
36
in cell division kinetics.
The incidence of radiation-induced MN in control samples was
REFERENCES
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37
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D
(
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3
4
28, (AlCl + HCl), 278, 390, (NaOAc), 266, 274, 350; (NaOAc +
3
H BO ), 264, 290sh, 372; IR ν 3367, 2936, 1680, 1622, 1565, 1511,
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3
3
max
−1 1
1
478, 1395, 1346, 1257, 1217, 1128, 1101, 1072, 1019, 828 cm ; H
13
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H] (calcd for C H O −H 637.1410).
28
30 17
2
2
Atriplexin II (2): yellow powder; [α] −82 (c 1, MeOH); UV λmax,
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D
(
(
MeOH) 240, 270, 346 nm, 270, 392 (NaOMe), 244, 268, 286, 382
AlCl ), 244, 268, 286, 378 (AlCl + HCl), 260, 278, 346 (NaOAc),
3
3
2
1
8
60, 278, 348 (NaOAc + H BO ); IR ν 3428, 2936, 1681, 1621,
3 3 max
566, 1514, 1478, 1392, 1346, 1257, 1217, 1128, 1101, 1072, 1019,
−1
1
13
28, 799 cm ; H NMR and C NMR data, see Table 1; HRESIMS
−
m/z: 651.1615 [M − H] (calcd for C H O −H 651.1567).
29
32 17
F
J. Nat. Prod. XXXX, XXX, XXX−XXX