Content uploaded by Felisa Parmar
Author content
All content in this area was uploaded by Felisa Parmar on Apr 28, 2021
Content may be subject to copyright.
IN VITRO ANTIOXIDANT AND ANTICANCER ACTIVITY OF MIMOSA PUDICA LINN EXTRACT
AND L-MIMOSINE ON LYMPHOMA DAUDI CELLS
Original Article
FELISA PARMAR, NISHA KUSHAWAHA, HYACINTH HIGHLAND, LINZ-BUOY GEORGE
Department of Zoology, Biomedical Technology and Human Genetics, University School of Sciences, Gujarat University, Ahmedabad
380009, Gujarat, India
Email: felisaparmar@yahoo.com
Received: 06 Aug 2015 Revised and Accepted: 27 Oct 2015
ABSTRACT
Objective: The present study is an attempt to investigate the antioxidant and anticancer potential of hydroalcoholic extract of Mimosa pudica Linn
(Mimosaceae) and L-Mimosine on Daudi cell line.
Methods: The analysis of the standard compound L-Mimosine was ascertained by High Performance Thin Layer Chromatography (HPTLC). Free
radical scavenging activity of M. pudica extract and L-Mimosine was also compared using 2, 2-diphenyl–1-picrylhydrazylradical scavenging assay
(DPPH). Cell viability and cytotoxicity on Daudi cells were evaluated by trypan blue and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium
bromide (MTT) assays in a dose and time-dependent manner respectively.
Results: HPTLC analysis showed the presence of amino acids, amines, lipids in the hydroalcoholic extract of M. pudica. Crude hydroalcoholic extract
of M. pudica showed antioxidant activity (IC50=103.88 µg/ml) whereas L-Mimosine showed antioxidant activity (IC50=233.06 µM). The crude
extract and compound inhibited the proliferation and growth of the Daudi cells through induced cell death. The IC50
Conclusion: The results indicated the presence of L-Mimosine in the hydroalcoholic extract of M. pudica. The hydroalcoholic extract and pure
compound proved potent inhibitors of cell proliferation, thus manifesting significant antioxidant and anticancer activity.
value for anticancer activity
was found to be 201.65 µg/ml and 86.61 µM at 72 h for M. pudica extract and L-Mimosine respectively.
Keywords: Anticancer, Antioxidant, Daudi, Mimosa pudica, L-Mimosine, HPTLC.
INTRODUCTION
Cancer has been one of the most dreaded diseases of the 20th century
and is increasing rampantly with greater intensity in the 21st
Over the last few decades, there has been increased interest by
pharmaceutical industries to discover new drugs from the
ethnobotanicals to provide new and alternative drugs to synthetic
drugs for the treatment of dreadful diseases. Only a few plants have
been significantly explored regarding their medicinal uses.
century. Cancer in adolescent and young adults is 2.7 times more
common than cancer occurring during the first 15 y of life. In
females aged between 15 to 29 y, malignancies of the genital tract
are the most frequent type of cancer (18%), followed by lymphoma
(17%) [1].
Antitumor drug resistance and side effects of antitumor compounds
are the most common hurdles in medicine [2]. Plants are rich
sources of anticancer agents and their derivatives are very useful for
the treatment or prevention of cancer. These plant compounds such
as vinblastine have been demonstrated as therapeutic for Hodgkin’s
lymphoma and vincristine for non-Hodgkin’s lymphoma while
Camptotheca acuminate, topotecan and irinotecan have been used
for lung and ovarian cancer [3]. Anticancer activity is the effect of
natural and synthetic or biological and chemical agents to reverse,
suppress or prevent carcinogenic progression. Several synthetic
agents are used to cure the disease but they have their cytotoxicity
and hence the research is now focused towards investigating plant
derived chemotherapeutic agents.
Mimosa pudica known as “Chue Mue” belongs to taxonomic group
Magnoliopsida and family Mimosaceae, is a stout straggling prostrate
shrubby plant, with compound leaves that get sensitive on touching,
pinous stipules and globose pinkish flower heads; it grows as a weed
in almost all parts of the country. Leaves and stems of the plant have
been reported to contain an amino acid Mimosine. The leaves also
contain mucilage and roots contain tannins [4]. M. pudica leaves are
used for its antihyperglycaemic, antiulcer, antidiarrheal,
anticonvulsant, cytotoxic and hepatoprotective properties. Moreover,
Paranjpe (1989) has reported that in Ayurvedic and Unani medicine,
M. pudica root is used to treat bilious fever, piles, jaundice, leprosy,
dysentery, vaginal and uterine complaints, inflammations and burning
sensation, fatigue, asthma, leucoderma and blood diseases [5].
L-Mimosine or leucenol [C8H10 N2O4
The cell line used in this study was Daudi cells which were cultured
from a line of lymphoblastoid cells derived from a human Burkitt
(non-Hodgkin) lymphoma which is cancer of the lymphatic system.
We have studied the effect of hydroalcoholic extract of M. pudica and
L-Mimosine on Daudi cells in vitro.
, β-N-(3-hydroxy-4-pyridone)-
amino propionic acid] is an amino acid present in plants of genus
Mimosa and Leucaena [6]. This rare amino acid exhibits a wide
range of effects, including inhibition of folate metabolism [7],
inhibition of deoxyribonucleotide metabolism [8], induction of
apoptosis [9, 10], inhibition of different cell line proliferation and in
vivo inhibition of tumor growth [11, 12]. It has shown an inhibitory
effect on different mammalian enzymes such as tyrosinase,
dopamine β-hydroxylase and deoxyhypusine hydroxylase [13, 14].
The aim of the current study is therefore to identify natural
compounds that can be used for prevention or treatment of cancer.
MATERIALS AND METHODS
Chemicals
L-Mimosine was purchased from MP Biomedicals (France) whereas all
other chemicals were purchased from Hi-Media Laboratories (India).
Preparation of plant extracts
The plant of M. pudica was collected from Gujarat University campus
and authenticated by Dr. H. A. Solanki, Professor, Botany
Department, Gujarat University, Ahmedabad. The whole plant viz.,
stems, leaves, roots and flower buds of M. pudica were collected,
washed and shade dried under ambient temperature. After complete
drying plants were powdered and defatted with petroleum ether
(40-60 °C) and kept for 24 h at room temperature with constant
shaking. 50 gm of the defatted powdered material was capsulated in
International Journal of Pharmacy and Pharmaceutical Sciences
ISSN- 0975-1491 Vol 7, Issue 12, 2015
Innovare
Academic Sciences
Parmar et al.
Int J Pharm Pharm Sci, Vol 7, Issue 12, 100-104
101
filter paper and kept in the thimble, 500 ml solvent (water: ethanol)
(70:30) was added into the flask and continuous extraction was
carried out in the Soxhlet apparatus for 72-74 h at 60 °C, (till the
colour in the siphon became colourless). The crude solvent collected
in the flask was dried at reduced pressure and kept at 4 °C until
further use.
DPPH radical scavenging activity
The antioxidant activity of plant extract and L-Mimosine was
measured in vitro using DPPH, a stable free radical. The reaction
mixture contained 0.1 ml of 0.1 mM DPPH and 0.1 ml of M. pudica
extract (31.25-250 µg/ml) or L-Mimosine (31.25-250 µM). The
solution was mixed rapidly and allowed to stand for 30 min in the
dark. The scavenging activity was measured by noting the decrease
in absorbance at 520 nm as compared to DPPH control. The analysis
was done in triplicate. The assay procedure was followed from those
described by Blois (1958) and Yamasaki et., al (1994)[15,16].
Inhibition of free radical DPPH in percent (I %) (or) the DPPH free
radical scavenging activity (%) was calculated from the absorption
according to the following equation:
I (%) (Or) DPPH Scavenged (%)
=Absorbance of control - Absorbance of sample
Absorbance of control ×100
Dose response curve was plotted between % inhibition and
concentrations. IC50
HPTLC
values were found out for plant extract as well
as for compound.
The extract was first run along with the standard compound on Thin
Layer Chromatography (TLC) plate through Phenol-Ethanol-Water
(3:1:1) and spots were developed by ninhydrin spraying reagent
(3% in acetone) [17]. This was then followed by scanning,
determination of retention factor (Rf) value and λ Max profile [18]
by Camag-5 HPTLC system WinCATS evaluation software (version
1.4.6.8121).
Cancer cell culture
For cancer cell culture, Daudi cell line was obtained from the
National Centre for Cell Science (NCCS), Pune. Cells were cultured in
RPMI 1640 media with 10% fetal bovine serum (FBS) and antibiotic-
antimycotic solution. Cell cultures were maintained in a CO2
incubator at 5% CO2
Cell viability assays by trypan blue dye exclusion technique
and 37 °C.
Any compound, which is cytotoxic to cells, inhibits the cell growth
proliferation and kills the cells. Trypan blue is a supravital dye, used
to estimate the number of cells present in the population [19]. It has
the ability to penetrate dead cells and give it a blue colour. This
method gives a score of dead and viable cells [20].
Cellular cytotoxicity induced by the M. pudica extract and L-
Mimosine was measured with trypan blue exclusion assay. Sterility
was maintained throughout the procedure. Briefly, 2×106
% Viability =Live cell count
Total cell count×100
cells were
seeded into 24-well plates and treated with or without (as control)
crude hydroalcoholic extract of M. pudica (12.5-400 µg/ml) and L-
Mimosine (12.5-400 µM) for 24, 48 and 72 h. After the incubation
period, the cultures were harvested and washed twice with
Phosphate Buffered Saline (PBS). The cell pellet was then
resuspended with 0.5 ml PBS. Then, 20 μL of cells were mixed with
equal volume of 0.4% trypan blue and was counted using a
Neubauer hemocytometer by clear field microscopy. Viable and non-
viable cells were counted. The percentage cytotoxicity was
calculated using the equation shown below:
In vitro cytotoxicity determination by MTT assay
The ability of cells to survive a toxic insult is the basis of most
cytotoxic assays. This assay is based on the assumption that dead
cells or their products do not reduce tetrazolium. It is described by
the modified method of Mosmann, (1983) and Wilson et al., (2000)
[21, 22].
The assay detects the reduction of MTT by mitochondrial
dehydrogenase to blue formazan product, which reflects the normal
function of mitochondria. 2×106 viable cells/ml were plated into the
96-well cell culture plate. The crude extract was added with the
concentrations (12.5-400 µg/ml) and compound with (12.5-400 µM)
respectively for 24, 48 and 72 h and incubated at 37 °C. After
incubation, the supernatants were removed and incubated with MTT
(0.5 v/v) in RPMI 1640 without FBS for 4 h in a humidified
atmosphere at 37 °C and 5% CO2
The absorbance (A) of the coloured solution was quantified at 540 nm
wavelengths by an enzyme-linked immunoabsorbent assay reader
(ELISA READER, MERCK MIOS mini). Each extract, compound and
control was assayed in triplicate in three independent experiments.
Percent growth inhibition of cells exposed to treatments was
calculated as follows:
% Inhibition
=100 Corrected mean Absorbance of sample
Corrected mean Absorbance of control
×100
incubator.
Statistical analysis
Each parameter was performed in triplicate and the results were
expressed as mean±standard error. The data was statistically
analyzed by Student’s ‘t’ test and the values of p<0.05 were
considered statistically significant.
RESULTS
Antioxidant effect M. pudica and L-Mimosine
The DPPH radical scavenging activity of extract and L-Mimosine is
shown in the fig. 1. It was observed that DPPH free radical scavenging
activity was concentration dependent in both the cases and reaches a
maximum at a concentration of 250 µg/ml for plant extract and 250
µM for the compound. Our result indicated that M. pudica extract
exhibited high antioxidative activity compared to L-Mimosine.
Fig. 1: DPPH scavenging activity of Mimosa pudica with IC50 value
of 103.88 µg/ml andL-Mimosine with IC50
Values are mean±S.E. for three individual experiments.
value of 233.06 µM
HPTLC
The peak heights area of the respective L-Mimosine standard and
the hydroalcoholic plant extract are shown in fig. 2 and 3. The
hydroalcoholic plant extract showed ten prominent spots on the TLC
plate with a retention factor of 0.09, 0.12, 0.21, 0.29, 0.45, 0.52, 0.62,
0.67, 0.84 and 0.94 (fig.3). HPTLC determination showed the
presence of amino acids, amines, lipids in the hydroalcoholic extract
of M. pudica and it was confirmed from the chromatogram after
derivatization with Ninhydrin spraying reagent (fig. 4).
Parmar et al.
Int J Pharm Pharm Sci, Vol 7, Issue 12, 100-104
102
Fig. 2: HPTLC chromatogram area and peaks of the L-
Mimosine using Win CATS evaluation software (version
1.4.6.8121)
Fig. 3: HPTLC chromatogram area and peaks of the Mimosa
pudica extract using Win CATS evaluation software (version
1.4.6.8121)
Fig. 4: Thin layer chromatogram of Mimosa pudica and L-
Mimosine using the solvent system Phenol: Ethanol: Water
(3:1:1 v/v/v) under 254 nm (A) and visible light (B)
Cell viability assay
The cell viability assay conducted by trypan blue dye exclusion
method showed that there was a highly significant (p<0.001)
decrease in viability with an increase in time and concentration in
both the extract as well as pure compound treated Daudi cells as
compared to untreated controlled cells.(fig. 5 and 6).
Fig. 5: Effect of Mimosa pudica extract on percentage viability on
Daudi cells. Values are mean±SE for three individual
experiments
Fig. 6: Effect of L-Mimosine on percentage viability on daudi cells.
Values are mean±SE for three individual experiments. Except for
treatment with 12.5 µM and 25 µM, there is a highly significant
(p<0.001) decrease of cell viability in all treated cells
Fig. 7: Effect of Mimosa pudica extract on the percentage
decrease of growth proliferation in Daudi cells. Values are
mean±SE for three individual experiments. Highly significant
(p<0.001) increase in growth inhibition was observed in all the
treated samples
MTT assay
Daudi cells were grown in 96 well plates for 24, 48 and 72 h with
different concentration (12.5-400 µg/ml) of the crude plant extract
and (12.5-400 µM) L-Mimosine compound. The formazan crystals
were formed, following the reduction of MTT by metabolically active
Parmar et al.
Int J Pharm Pharm Sci, Vol 7, Issue 12, 100-104
103
(viable) cells. The percentage decrease of proliferation after
treatment with the M. pudica and L-Mimosine is given in Fig.7 and 8
respectively compared to control. The IC50
value was calculated by
plotting the graph in Microsoft Excel and was found to be 201.65
µg/ml and 86.61 µM at 72 h for M. pudica extract and L-Mimosine
respectively. There was a significant increase in the percentage of
inhibition of growth proliferation with increased dose and time
duration as compared to untreated control cells (p<0.001).
Fig. 8: Effect of L-Mimosine extract on the percentage decrease of
growth proliferation in Daudi cells. Values are mean±SE for three
individual experiments. Highly significant (p<0.001) increase in
growth inhibi tion was observed in all the treated samples
DISCUSSION
The greatest challenge for phytochemical and pharmacological
studies involves the identification of the specific compounds that are
responsible for the beneficial effects and their modes of action,
thereby delineating their useful functions as therapeutic drugs.
Mimosa pudica Linn is a well-known herbal medicine throughout the
world. Many studies have reported the pharmacological efficacies
and benefits of M. pudica. Jose et., al. (2014) have shown potential
anticancer activity with isolated flavonoids of M. pudica against
MCF-7, human breast cancer cell line. Cytotoxic activity of M. pudica
was evaluated using brine shrimp lethality bioassay which had
shown little cytotoxic effect and results suggest that the plant can be
used as a promising source of anticancer compounds [24]. There is
very little information available on its mechanism of action on
cancer cells. On the other hand, Lalande and Hanauske-Abel (1990)
have shown that Mimosine induces cell-arrest (reversibly) late in the
G1 phase of the cell cycle.
Moreover, Mimosine is known to be a tyrosine analog that contains a
metal chelating domain and hence it is able to chelate transition
metals, such as Fe3+
DPPH has been used to evaluate the free radical scavenging activity
of the natural antioxidant. DPPH which is a radical itself with a
yellow color, changes into a stable compound antioxidant and the
extent of the reaction depends on the hydrogen donating ability of
the antioxidant [34]. Both extract and compound showed potent free
radical scavenging activity with IC
(Linn et al. 1996). The possible structure of the
Fe (III)-Mimosine chelation complex has also been proposed by Tsai
and Ling (1973). Furthermore, there is evidence that Mimosine
inhibits various mammalian enzymes in vitro, such as tyrosinase,
dopamine hydroxylase [13], deoxyhypusyl hydroxylase (DOHH)
[28], and H1 kinase [29]. These results strongly suggest that
Mimosine may inhibit the intense mitotic activity of cancer cells. In
fact, in vitro studies have demonstrated that Mimosine represses
uterine cancer cell growth [30] and blocks DNA replication in both
breast cancer and Chinese hamster ovary cells [31]. It has been
shown that Mimosine acts as an iron chelator [32] and the resulting
iron deficiency could alter folate metabolism in mammals and
interfere with tumor cell growth [33]. The DPPH assay is based on
the reduction of stable radical DPPH to yellow coloured diphenyl
picryl hydrazine. Thus, the ability of the tested products to quench
this radical is a measure of its antioxidative ability. Many previous
studies have reported strong to the moderate free radical activity of
crude extracts of the plants belonging to Mimosaceae. Normal cells
have been studied previously in the same experiment and no
significant effect has been found on normal cells.
50
In this study, hydroalcoholic extract of M. pudica and L-Mimosine
were used to evaluate their possible anticancer activity. Both M.
pudica hydroalcoholic extract and L-Mimosine treatment caused a
significant loss of viability of cells as measured by this assay in a
dose and time-dependent manner respectively.
value of 103.88 µg/ml and
233.06 µM respectively. The ability of the extract of M. pudica and L-
Mimosine to scavenge DPPH radicals suggests that it can react with
free radicals to convert them to more stable products and terminate
radical chain reaction.
The cytotoxic activity was monitored by the standard MTT assay.
The ability of cells to survive a toxic insult is the basis of most
repeated cytotoxic assays. This assay depends on both on the
mitochondrial activity per cell and number of cells present. The
cleavage of MTT to a blue formazon derivative by living cells is
clearly a very effective principle on which the assay is based
conforming the decrease in survival of the cultured cells.
The loss of viability and anticancer effect of L-Mimosine on Daudi cell
line is might be due to inhibition of cellular enzymes in vitro [30].
CONCLUSION
The present study reported L-Mimosine from Mimosa pudica.
Present data indicate that L-Mimosine treatment exhibited less
antioxidant activity. Treatment with both hydroalcoholic extracts of
M. pudica/ L-Mimosine showed potent anticarcinogenic effects
against Daudi Lymphoma cell line leading to anti proliferation and
loss of cell viability. To conclude data suggested that L-Mimosine
treatment exhibited much higher anticarcinogenic activity compared
to the extract of M. pudica.
CONFLICT OF INTERESTS
The authors have declared no conflict of interests.
REFERENCES
1. Bleyer A, Viny A, Barr R. Cancer in 15-to 29-year-olds by
primary site. Oncologist 2006;11:590-601.
2. Holohan C, Schaeybroeck SV, Long ley DB, Johnston PG. Cancer
drug resistance: an evolving paradigm. Nat Rev Cancer
2013;13:714-26.
3. Cragg GM, Newman DJ. “Plants as a source of anticancer
agents,”. J Ethnopharmacol 2005;100:72-9.
4. Ghani A. Medicinal plants of Bangladesh with chemical
constituents and uses. ed 2. Dhaka, Bangladesh; 2003. p. 302-3.
5. Paranjpe P. Indian medicinal Plants: Forgotten healers. Delhi,
India; 1989. p. 155-6.
6. Thompson JF, Morris CJ, Smith IK. New naturally occurring
amino acids. Annu Rev Biochem 1969;38:137-58.
7. Oppenheim EW, Nasrallah IM, Mastri MG, Stover PJ. Mimosine
is a cell-specific antagonist of folate metabolism. J Biol Chem
2000;275:19268-74.
8. Gilbert DM, Neilson A, Miyazawa H, De Pamphilis ML, Burhans
WC. Mimosine arrests DNA synthesis at replication forks by
inhibiting deoxyribonucleotide metabolism. J Biol Chem
1995;270:9597-606.
9. Reno F, Tontini A, Burattini S, Papa S, Falcieri E, Tarzia G.
Mimosine induces apoptosis in the HL60 human tumor cell line.
Apoptosis 1999;4:469-77.
10. Zalatnai A, Bocsi J. Mimosine, a plant-derived amino acid
induces apoptosis in human pancreatic cancer xenografts.
Anticancer Res 2003;23:4007-9.
11. Chang HC, Weng CF, Yen MH, Chuang LY, Hung WC. Modulation
of cell cycle regulatory protein expression and suppression of
tumor growth by mimosine in nude mice. Int J Oncol
2000;17:659-65.
12. Ju H, Hao J, Zhao S, Dixon IM. Antiproliferative and antifibrotic
effects of mimosine on adult cardiac fibroblasts. Biochim
Biophys Acta 1998;1448:51-60.
13. Hashiguchi H, Takahashi H. Inhibition of two copper-containing
enzymes tyrosinase and dopamine beta hydroxylase, by L-
mimosine. Mol Pharmacol 1977;13:362-7.
Parmar et al.
Int J Pharm Pharm Sci, Vol 7, Issue 12, 100-104
104
14. Hanauske-Abel HM, Park MH, Hanauske AR, Popowicz AM,
Lalande M, Folk JE. Inhibition of the G1-S transition of the cell
cycle by inhibitors of deoxyhypusine hydroxylation. Biochim
Biophys Acta 1994;1221:115-24.
15. Blois MS, Zhao XY. Antioxidant determinations by the use of
stable free redicals. Nature 1985;181:1199-200.
16. Yamasaki K, Hashimoto A, Kokusenya Y, Miyamoto T, Sato T.
Electrochemical method for estimating the antioxidative effects
of methanol extracts of crude drugs. Chem Pharm Bull
1994;42:1663-5.
17. Spenser ID, Notation AD. A synthesis of mimosine. Can J Chem
1962;40:1374-79.
18. Sharma A, Patel VK. In vitro screening of the antibacterial
activity and identification of bioactive compounds from plants
against selected Vibrio spp. pathogens. Turk J Biol
2009;33:137-44.
19. Phillips HJ, Terryberry JE. Counting actively metabolizing tissue
cultured cells. Exp Cell Res 1957;13:341-7.
20. Kuttan R, Bhanumathly P, Nirmala K, George MC. Potential
anticancer activity of turmeric (Curcuma longa). Cancer Lett
1985;29:197-202.
21. Mosmann T. Rapid colorimetric assay for cellular growth and
survival: application to proliferation and Cytotoxicity assays. J
Immunol Methods 1983;65:55-63.
22. Wilson AP. Cytotoxicity and viability Assay in Animal Cell culture,
A Practical Approach. ed 3. New York, NY; 2000. p. 175-219.
23. Jose J, Sudheesh S, Sumesh Kumar TM, Sony J, Jayadevi Variyar E.
A comparative evaluation of anticancer activities of flavonoids
isolated from Mimosa pudica, Aloe vera, Phyllanthus niruri against
human breast carcinoma cell line (MCF-7) using MTT assay. Int J
Pharm Pharm Sci 2014;6:319-22.
24. Srikanta C, Dibyajyoti S, Swati P. In vitro cytotoxic activities of
methanolic extracts of Mimosa pudica. Bull Pharm Res
2012;2:42-5.
25. Lalande M, Hanauske-Abel HM. A new compound which
reversibly arrests T lymphocyte cell cycle near the G1/S
boundary. Exp Cell Res 1990;188:117-21.
26. Linn HB, Falchetto R, Mosca PJ, Shabanowitz J, Hunt DF, Hamlin
JL. Mimosine targets serine hydroxymethyltransferase. Biol
Chem 1996;271:2548-56.
27. Tsai WC, Ling KH. Study on the stability constant of some metal
ion chelatea of mimosine and 3,4-tihydroxypyridine. J Chin
Biochem Soc 1973;2:70-86.
28. Abbruzzese A, Hanauske AHM, Park MH, Henke S, Folk JE. The
active site of deoxyhypusyl hydroxylase: use of catechol
peptides and their component chelator and peptide moieties as
molecular probes. Biochim Biophys Acta 1991;1077:159-66.
29. Feldman ST, Schonthal A. Negative regulation of histone H1
kinase expression by mimosine, a plant amino acid. Cancer Res
1994;54:494-8.
30. Zalatnai A. P-glycoprotein expression is induced in human
pancreatic xenografts during treatment with a cell cyclo
regulator, mimosine. Pathol Oncol Res 2005;11:164-9.
31. Mosca PJ, Dijkwel PA, Hamlin JL. The plant amino acid
mimosine may inhibit initiation at origins of replication in
Chinese hamster cells. Mol Cell Biol 1992;12:4375-83.
32. Kulp KS, Vulliet PR. Mimosine blocks cell cycle progression by
chelating iron in asynchronous human breast cancer cells.
Toxicol Appl Pharmacol 1996;139:356–64.
33. Gilbert DM, Neilson A, Miyazawa H, Depamphilis ML, Gupta HK,
Altreja PP. Influence of ferric chloride treated Leucaena
leucocephala on metabolism of mimosine and 3-hydroxy-4
(1H)-pyridone in growing rabbits. Anim Feed Sci Technol
1998;74:45-55.
34. Chen CW, Ho CT. Antioxidant properties of po lyphenols
extracted from green and black teas. J Food Lipids
1995;2:35-46.
