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Cherry (Prunus Cerasus) is still one of the most popular preserve in Turkish cuisine. Cherry has been traditionally used for the treatment of inflammatory-related symptoms. Recent researches have proved that cherry is a valuable natural source of some important bioactive compounds in human health preservation. Evidence suggests that, cherry consumption may decrease the risk of chronic diseases and cancer. The aim of the present study was to determine the effects of cherry on breast cancer cells lines, asymmetric dimethylarginine (ADMA) level and certain multidrug-resistant bacteria. The cancer cell proliferation activity and analysis of apoptotic-necrotic cells was evaluated by using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and scoring of apoptotic cell nuclei. Measurement of ADMA and the minimum inhibitory concentration was accomplished by HPLC and the micro dilution broth method. The results showed that, extracts of cherry exhibit anti-proliferative activity in mammary adenocarcinoma (MCF-7) & mouse mammary tumor cell (4T1) breast cancer cells lines as well as induction of apoptosis, lower ADMA concentrations in cell cultures treated with cherry extract and antibacterial effects against certain multidrug-resistant bacteria in vitro. These findings may open new horizons for traditional anti-inflammatory product as prophylactic-therapeutic agent from cancer, cardiovascular diseases and multidrug-resistant infections.
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Pak. J. Pharm. Sci., Vol.27, No.3, May 2014, pp.587-592 587
REPORT
Investigation of anti-cancer effects of cherry in vitro
Recai Ogur1*, Hakan Istanbulluoglu2, Ahmet Korkmaz3, Asli Barla4,
Omer Faruk Tekbas1 and Emin Oztas5
1Department of Public Health, Division of Environmental Health, Gulhane Military Medicine Faculty, Etlik, Ankara, Turkey
2Department of Public Health, Gulhane Military Medicine Faculty, Etlik, Ankara, Turkey
3Department of Physiology, Gulhane Military Medicine Faculty, Etlik, Ankara, Turkey
4Extraction Department Responsible, AROMSA Co., Istanbul, Turkey
5Department of Histology and Embriology, Gulhane Medical Faculty, Ankara, Turkey
Abstract: Cherry (Prunus Cerasus) is still one of the most popular preserve in Turkish cuisine. Cherry has been
traditionally used for the treatment of inflammatory-related symptoms. Recent researches have proved that cherry is a
valuable natural source of some important bioactive compounds in human health preservation. Evidence suggests that,
cherry consumption may decrease the risk of chronic diseases and cancer. The aim of the present study was to determine
the effects of cherry on breast cancer cells lines, asymmetric dimethylarginine (ADMA) level and certain multidrug-
resistant bacteria. The cancer cell proliferation activity and analysis of apoptotic-necrotic cells was evaluated by using
the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and scoring of apoptotic cell nuclei.
Measurement of ADMA and the minimum inhibitory concentration was accomplished by HPLC and the micro dilution
broth method. The results showed that, extracts of cherry exhibit anti-proliferative activity in mammary adenocarcinoma
(MCF-7) & mouse mammary tumor cell (4T1) breast cancer cells lines as well as induction of apoptosis, lower ADMA
concentrations in cell cultures treated with cherry extract and antibacterial effects against certain multidrug-resistant
bacteria in vitro. These findings may open new horizons for traditional anti-inflammatory product as prophylactic-
therapeutic agent from cancer, cardiovascular diseases and multidrug-resistant infections.
Keywords: Cherry, Breast Cancer Cells, Anticancer Effect, Minimum Inhibitory Concentration, ADMA.
INTRODUCTION
Cherry (Prunus cerasus) is a sour fruit strain that is like
berry from the rosaceae family. The region between the
Caspian Sea and North Anatolia is regarded as the
possible homeland of cherry in most of the resources.
Cherry especially is quite rich in terms of mineral matter.
By the reason of fruit juice’s efficiency’s (70-75%) and
total acidity’s (3%) being high cherry is very convenient
to be processed as fruit juice (Turkey Fruit Juice Industry
2011; Krishkov 2009). In Turkey the production of cherry
that is 24.000 ton in 1965 reached 85.000 ton in 1985 and
120.000 ton in 1991. In this aspect Turkey ranks as the
fifth country in cherry production. Differently from other
countries, in our country the mostly consumed fruit juices
are peach and cherry. The consumption of fruit juice that
is 0,4 liter per person in the beginnings of 1970s
increasing 10 times reached 3,9 liter per person in 1996
and today the rate is around 4,5 liter (Turkey Fruit Juice
Industry 2011).
According to the researches, antioxidant capacity of the
cherry is quite high and can be defined as nutriceuticals. It
has been denoted that anthocyanins in cherry have the
effects of decreasing rheumatism, the risk of colon cancer
and inhibiting fever, inflammation and tumor progression.
Cherry being rich in terms of melatonin that is a powerful
antioxidant and well sleeping regulator produced in the
brain backs up growth and cell renewal (Jafari et al.,
2008; Kim et al., 2005; Burkhardt et al., 2001; Wang et
al., 1999).
Nitric oxide (NO) is the most important endothelium
borne vasodilator. In addition to NO’s vascular smooth
muscle proliferation’s positive effects on thrombocyte
aggregation and vascular superoxide productions, it has
anti-atherosclerotic features. ADMA is the major inhibitor
of NO biosynthesis in human body. It is thought that the
increase of ADMA leading endothelial dysfunction has a
significant role in atherogenesis and cardiovascular
diseases (Vallance et al., 1992; Boger 2004).
Some bacteria like methicillin-resistant Staphylococcus
aureus, carbapenem-resistant Pseudomonas aeruginosa
and Acinetobacter baumannii developed resistance ways
against drugs’ effects. Along with surveying resistance
mechanisms and new drugs, these resistant
microorganisms, pose serious problems for health care
workers especially nosocomial infection at the top (Jeong
et al., 2006; Chastre and Trouillet, 2000).
*Corresponding author: e-mail: rogur@gata.edu.tr
Investigation of anti-cancer effects of cherry in vitro
Pak. J. Pharm. Sci., Vol.27, No.3, May 2014, pp.587-592
588
The purpose of this study is to investigate the potential
anticancer effects of cherry on human breast mammary
adenocarcinoma (MCF-7) and mouse mammary tumor
cell (4T1). To evaluate ADMA concentrations in cell
cultures treated with cherry extract and to examine
whether cherry has an antibacterial effect against certain
multidrug-resistant bacteria in vitro.
MATERIALS AND METHODS
Reagents
Maltodextrin Non-GMO was purchased from Tate & Lyle
Corporate and Investor Relations, Company, Slovakia.
Chemical and reagents; Folin-Ciocalteu reagent 2N,
DPPH (1,1-diphenyl-2-picrylhydrazyl), gallic acid, alpha
tocopherol, BHA (3-tert-butyl-4-hydroxyanisole) were
purchased from Sigma-Aldrich (St. Louis, MO) and
sodium carbonate was from Alfa Aesar GmbH & Co, KG.
All other chemicals were of food grade.
Plant material and preparation of extracts
Extracts of cherry pulps were used in the study. Cherry
pulp concentrations were provided by an international
aroma producer (Aromsa, Turkey). Cherry concentrate
was macerated with water and ethanol (1:1 w/w) for 4
times at room temperature. Evaporation of the solvent
under vacuum yielded 40 BX cherry extract, which used
in spray dry. Another concentrates from cherry
concentrates were used as a concentrate at 65 BX in all
tests.
Preparation of microcapsules by spray drying
Microencapsulation of flavors in carrier matrices can
provide protection prevent loss of volatile flavors and
enhance stability of the flavor core materials. The most
common and economical way to apply microencap-
sulation is spray drying. There are many papers were
published about the encapsulation of flavors in liquid by
spray drying. In this study, microencap-sulation was
performed by Aromsa Co. Inc and maltodextrin Non-
GMO used as a wall material. The total concentration of
cherry extract was 40% (w/w) and that was blended of
maltodextrin 30% (w/w; MD:Water). The mixture was
emulsified in a Sylverson, (Sylverson, Chesham,
England) for 5 min at 4000 rpm until complete dispersion
of the extract. The resulting slurry was spray dried in
APV-Anhydro, Denmark. The microcapsules prepared
were collected from the collecting chamber and filled in
air tight self-sealable polyethylene pouches.
Studied activity
Total phenolic content was measured by using the Folin-
Ciocalteu’s reagent. Results are expressed as microgram
of gallic acid equivalents per mg of extract. Free radical
scavenging activity was determined by 1,1-diphenyl-2-
picryl-hydrazil (DPPH) assay. The data on all antioxidant
activity tests were triplicated.
Cancer cell lines
Dulbecco's Modified Eagle Medium (DMEM) and fetal
bovine serum (FBS) was purchased from Invitrogen
Corporation (Carlsbad, CA 92008 USA). Dimethyl
sulphoxide (DMSO), phosphate buffer saline (PBS
solution) and MTT were purchased from Sigma-Aldrich
Chemical Co. (St. Louis, MO). MCF-7 and 4T1 cell lines
were used, in order to determine antitumoral activity,.
Cancer Cell proliferation inhibitory assay
The cancer cell proliferation activity of the cherry
extraction was tested by MTT colorimetric assay. Cancer
cell cultures (MCF-7 & 4T1) were maintained at the
Public Health Laboratory at Gulhane Medicine Faculty
Ankara, Turkey. MCF-7 & 4T1 were cultured in 10%
FBS, 2 µM L-glutamine, 100 µg/ml streptomycin and 100
µg/ml penicillin were added in DMEM medium in a
humidified incubator at 37°C under 5% CO2. MCF-7 &
4T1 cell lines were harvested, counted and moved into
96-well plates and incubated for 24 hours before
treatment. By melting away the cherry extracts in DMSO
the samples were made ready. Samples at 50 and
100µg/mL, were supplemented to DMEM medium. The
ending concentration of DMSO in the assay was 0.1%.
Samples at 50 and 100µg/mL concentrations were
supplemented to the wells. Into each well 25/µL MTT
solution was added after 48 hours incubation of samples
with cancer cells. Plates were incubated for 3 h at 37°C.
For dissolving the formed formazan crystals, content of
each well removed and 200/µL DMSO was added to each
well, and then the plates were shaken. Plates were
incubated for 5 min at room temperature, and finally the
optical density (OD) of the wells was ascertained using a
micro-plate reader at 570 nm. Samples' cell viability at
each concentration was calculated related to solvent
control. At each concentration, cell viability was
evaluated by dividing the optical density of samples with
the optical density of solvent control.
Analysis of apoptotic and necrotic cells
After treating cell lines with different amounts cherry
extract for 72 hours, all of the cells (attached and
detached cells) in the flask were brought together. After
this step, cells were washed with PBS for three times and
stained with Hoechst dye (2mg/ml), propodium iodide
(PI) (1mg/ml) and DNAse free-RNAse (100mg/ml) at the
25°C for 15 minutes and analyzed under a confocal
fluorescence microscope (LSM-GB200: Olympus, Tokyo,
Japan). The nuclei of apoptotic cells were marked blue by
Hoechst dye where necrotic cells were marked red by PI.
Ten areas in microscope, chosen by chance, were counted
and the result presented as a ratio of apoptotic and
necrotic to normal cells.
Measurement of ADMA
Measurement of ADMA was performed by HPLC (high
pressure liquid chromatography), refer to the method
described by Chen et al., (Chen et al., 1997). In short, 1
Recai Ogur et al.
Pak. J. Pharm. Sci., Vol.27, No.3, May 2014, pp.587-592 589
ml cell culture medium and 20/mg 5-sulfosalicylic acid
was mixed and the mixture was kept waiting in an ice
bath for 10 min. After centrifugation for 10 min at 2000
rpm the precipitated protein was removed. 10/µl of the
supernatant that was filtered and mixed with 100/µl of
extraction reagent (10/mg o-phthaldialdehyde in 0.5/ml of
methanol and 2/ml of 0.4/m borate buffer) then injected
into HPLC.
The minimum inhibitory concentration
Antibacterial characteristic of cherry evaluated on two
standard (Staphylococcus aureus ATCC 29123,
Pseudomonas aeruginosa ATCC 27853) and three clinical
(MRSA, Carbapenem-resistant Acinetobacter baumannii,
Carbapenem-resistant Pseudomonas aeruginosa) strains
of bacteria. The micro dilution broth method was used for
carrying out the minimum inhibitory concentration
(MIC). By using Mueller Hinton Broth sequential two
times dilutions of cherry extract were done in sterile 96-
well micro-plates with concentrations between ½ and
1/1024. Bacterial suspensions were regulated to 0.5 Mc
Farland standards. From an agar plate culture, at least
three well-separated colonies were chosen and they were
incubated at 37°C for 48 hr. Bacterial growth was
analyzed by 625 nm. absorbance measurement after
incubation. DMSO was used as a control. Ascertain the
late antimicrobial activity of cherry all assays were also
incubated for 48 hr at 37°C aerobically. Bacterial growth
was analyzed by 625 nm. absorbance measurement after
incubation. Lowest concentration of cherry which limited
bacterial growth to a <0.05 level at 625 nm was accepted
as MIC.
STATISTICAL ANALYSIS
Statistical analysis of variance was determinate by
ANOVA, significant differences between means were
determined by Student’s-t test, P<0.05 were noted as
significant.
RESULTS
Activity result
The cherry extract encapsulated was found to be the most
active extract in tests. Previously, the cherry extract
encapsulated was found to be rich in phenolic
compounds. Total phenolic content of cherry
encapsulated extracts were varied between 93.7 mg and
115.6 mg (mg GEs/100 g extract). Total phenolic content
of the other cherry concentrates were varied between 54.3
mg and 79.0/mg (mg GEs/100 g extract). As far as DPPH
assay, the cherry extract encapsulated was showed higher
antioxidant activity than Ascorbic acid and all tested
extracts of cherry (table 1). Konya and Tokat Cherry
concentrate were showed similar activity in DPPH assay
and all other extracts showed higher activity of Ascorbic
acid (fig. 1).
Results of MTT
The cell proliferation inhibitory effects of encapsulated
cherry extracts that have 50 and 100ug/ml concentration
respectively has been examined in the MCF-7 and 4T1
cancerous cell series. It has been concluded that the
extracts that are added to cell culture not only decreased
the cell proliferation in both MCF-7 and 4T1 cells but
also decreased in the concentration. It has been stated that
above mentioned extracts the one that has 50 ug/ml
concentrations inhibited cell growth 37% in MCF-7 cell
series and 48% in 4T1 series. Similarly, it has been stated
that the extract that has 100 ug/ml concentration inhibited
cell proliferation 54% in MCF-7 series and 53% in 4T1
series. It has been specified that the decrease ascertained
in the proliferation in cell series were significance
statistically (fig. 2).
DPPH ASSAY RESULTS
0,00
10,00
20,00
30,00
40,00
50,00
60,00
70,00
80,00
90,00
100,00
BHA
Ascorbic acid
Sour cherry extract Encapsulated
Kütahya Sour cherry Concentrate
Konya Sour cherry Concentrate
Tokat Sour cherry Concentrate
Afyon Sour cherry Concentrate
Sour Cherry Extracts and Standarts
% INHIBITION (IC5 0
)
Series1
Fig. 1: DPPH assay results of cherry extracts.
0
20
40
60
80
100
120
050100
Conc. (µg/mL)
% Viability
MCF-7
4T1
Fig. 2: In vitro cell proliferation inhibitory results of
cherry against cancer cell lines
Results of apoptotic and necrotic cells analysis
The effects of encapsulated cherry extracts on cell
development and its survival have been analyzed by
staining the MCF-7 and 4T1 cells that were treated with
cherry extract with Hoechst and Propodium Iodid. In
consequence of microscopic examination, it has been
stated that the greater part of examined cells cannot be
stained by PI and they are at late apoptosis phase that is
characterized by cells whose nucleuses are separated into
smaller organisms in cytoplasmic membrane. In addition
to these analyzed fields, the existence of red painted early
Investigation of anti-cancer effects of cherry in vitro
Pak. J. Pharm. Sci., Vol.27, No.3, May 2014, pp.587-592
590
and late necrotic cells have been specified. In the light of
findings above, it has been resulted that cherry extracts
induced anticancer activity through apoptosis (fig. 3).
Fig. 3: A general view of apoptotic and necrotic cells by
fluorescence microscopy. Blue points represent the
apoptotic cells, red points represent necrotic cells.
Table 1: Minimal inhibitory concentration (µL/mL)
results of cherry against clinical and standards strains of
bacteria.
24 hr of
incubation
(µL/mL)
48 hr of
incubation
(µL/mL)
Clinical isolates 200 100
Methicillin-resistant
Staphylococcus aureus
100 12.25
Carbapenem-resistant
A. baumannii
100 12.25
Carbapenem-resistant
Ps.aeruginosa
100 12.25
Standard strains
S. aureus ATCC 29123 200 100
Ps. aeruginosa ATCC 27853 100 12.25
Results of ADMA
ADMA levels of cell cultures supplemented with cherry
extract were significantly lower than the Ø control and
control cell culture values. ADMA levels of cell cultures
supplemented with cherry extract, Ø control and control
cell culture are 1.21, 2.54 and 2.79µmol/l respectively.
Minimal inhibitory concentration (µL/mL)
The results obtained from the MIC examination show that
cherry extract is effective on three clinical (MRSA,
Carbapenem-resistant Acinetobacter baumannii,
Carbapenem-resistant Pseudomonas aeruginosa) and two
standard (Staphylococcus aureus ATCC 29123,
Pseudomonas Aeruginosa ATCC 27853) examined strains
of bacteria. Following 24 hours incubation, MIC
concentration has been found 200ug/ml for gram-positive
microorganisms and 100ug/ml for gram-negative
microorganisms. Similarly, following 48 hours incubation
it has been stated that MIC rate fell 100ug/mg for gram-
positive microorganism and 12.25ug/ml for gram-
negative microorganism. At the same period there has
been obtained increase in the number of microorganisms
at the control wells.
DMSO, used as negative control, its effect on bacteria
proliferation has been researched. At the ½ diluted well of
Carbapenem-resistant Pseudomonas aeruginosa except
detected inhibition at the end of 48 hours incubation there
aren’t detected any inhibition in other wells. Including
positive controls, all of the results obtained from MIC
examination are in the border of CLSI quality control
parameter.
DISCUSSION
This is the first study in which the effect of cherry on
MCF-7 and 4T1 human and mouse breast cancer cell lines
was surveyed. The results obtained from this study is in
accordance with the results of a study in which the effects
of cyanine, giving the bright color of cherry and classified
as subgroup of anthocyanins, on human colon cancer was
surveyed. Furthermore, it has been stated that cherry
inhibits the formation and development of intestinal
cancer on ApcMin mice that are developed as human
cancer model. In the stated study, the number and size of
adenoma have been stated respectively less and small in
the mice whose diet cherry is added compared to other
mice (Kamei et al., 1998; Kang et al., 2003).
NAG-1 has been ascertained to be a gene that shows anti-
tumorigenic activity by triggering the apoptosis in the cell
lines. Studies proved that NAG-1 is a significant
promising target gene in inhibiting the cancer
development. The “wild-cherry’’ extract sometimes called
as “black-cherry’’ has been stated to increase NAG-1
expression and inhibit cell reproduction in the cancer cell
lines by increasing apoptosis as well (Baek et al., 2001;
Liu et al., 2003).
The surveys conducted revealed that wild-cherry extract
respectively suppresses B-catenin/TCF signal. It has been
stated that in the colorectal adenokarsinoms cyclin D1
protein expression increases. Similarly, the results of wild
cherry treatment made in the colorectal adenokarsinom
cell lines verify the decrease of cyclin D1 protein
expression with treatment. In the light of this information
it has been assessed that wild cherry suppresses B-catenin
signal in cancer cells and in consequence of this
suppression cyclin D1 protein expression decreases and
cancer cell development regresses (Yamaguchi et al.,
2006; Fu et al., 2004).
Genetic surveys states that genetic features of different
cherry kinds resemble each other very much. NAG-1
Recai Ogur et al.
Pak. J. Pharm. Sci., Vol.27, No.3, May 2014, pp.587-592 591
gene’s and cyclin D1 protein’s being covalent in the
analyzed kinds is possible. It has been thought that anti-
tumoral effect stated in this survey occurred by gene and
proteins in question (Mariette et al., 2010; Stockinge et
al., 1996).
When we consider the results of the survey, cherry is
specified to have shown antimicrobial action against the
gram-negative and gram positive bacteria examined in the
cope of survey. Examined bacteria’s causing nosocomial
infection and their being antibiotic resistant increases
importance of obtained results of the survey. Cherry along
with showing antimicrobial action on both gram-negative
and gram-positive bacteria, its effect on gram-negative
bacteria is stronger. In the previous studies cherry has
been mentioned to shown antibacterial action by
inhibiting the adhesion of bacteria. Mechanism of action
of the cherry is a subject that has not been sufficiently
researched yet. It is possible that the difference of the
action that cherry showed towards gram-negative and
gram-positive bacteria kind results from the difference of
the protein and lipid layer in the membrane structure of
these bacteria kinds. There is needed further study on this
subject (Klevens et al., 2007; Hebeler et al., 1973;
Rothfield et al., 1964).
In a recent study it is stated that much of fruit juices and
extracts especially those that are rich of anthocyanins and
acidic show antibacterial effect. It is ascertained that the
action in question disappears with the disappearance of
the acidic structure. However, in the same study it is also
ascertained that cherries and raspberries extracted with
water and blackcurrant extracted with methanol show
highly antibacterial effect. Effect showed in cherry,
raspberry and blackcurrant is stated to have been
independent from low acidic level and resulted from
anthocyanins and ellagitannin that are intensive in these
fruits (Galgóczy et al., 2009).
In this study ethanol was used in the preparation of cherry
extract. This situation made us think that the strong
antibacterial effect ascertained in cherry extracts, as
showed in some other studies, depends on both used
extract’s being acidic and pigments included in cherry’s
ingredient like anthocyanins and etc (Lee et al., 2003;
Harborne and Williams 2000).
This is the first study in which ADMA level increase
observed in MCF-7 and 4T1 breast cancer lines treated
with cherry extract. After showing its inhibiting NO
synthesis, in a short time lots of study was carried out
about ADMA’s pathophysiology (Valkonen et al., 2001;
Böger at al 2003). Increased ADMA levels by inhibiting
NO synthesis activity, inhibits NO formation and vascular
structure is preserved. In recent studies it is stated that
anthocyanins in cherry have positive effects on vascular
endothelium cells. It is expressed that mentioned effects
were controlled by plenty factors and enzymes partaking
in ADMA catabolism like Dimetilarginin dimetil amimo
hidrolaz are among these factors (Ding et al., 2000; Mc
Carty 2004).
The effects of cherry on cancer development, antibiotic-
resistant gram-positive and gram-negative pathogen
microorganisms and ADMA levels is of great concern to
human health because it is an ingredient in foods
consumed daily. Whereas, little is known about the
metabolic outcome of ingested cherry and its compounds
in humans in order to correlate its health benefits in vivo
as compared to in vitro studies.
ACKNOWLEDGEMENTS
This project was supported by the Aromsa Besin Aroma
ve Katki Maddeleri San ve Tic. AS and MEYED (Turkey
Fruit Juice Industry Association).
REFERENCES
Baek SJ, Kim KS, Nixon JB, Wilson LC and Eling TE
(2001). Cyclo-oxygenase inhibitors regulate the
expression of a TGF-beta super-family member that
has proapoptotic and antitumorigenic activities. Mol.
Pharmacol., 59: 901-908.
Boger RH (2004). Asymmetric dimethylarginine, an
endogenous inhibitor of nitric oxide synthase, explains
the ‘l-arginine paradox’ and acts as a novel
cardiovascular risk factor. J. Nutr., . 134(10): 2842-
2847.
Boeger RH, Lenzen H, Hanefeld C, Bartling A, Osterziel
KJ, Kusus M, Schmidt LC, Stroedter D, Berger J,
Goudeva L and Muegge A.. (all names of author are
required) (2003). Asymmetric dimethylarginine, an
endogenous inhibitor of NO synthase is a predictor of
the risk for coronary heart disease Results of the
multicenter CARDIAC study. Circulation, 108(4): 256.
Burkhardt S, Tan DX, Manchester LJ, Hardeland R and
Reiter RJ (2001). Detection and quantification of the
antioxidant melatonin in Montmorency and Balaton
tart cherries (Prunus cerasus). J. Agric. Food Chem.,
49(10): 4898-4902.
Chastre J and Trouillet JL (2000). Problem pathogens
(Pseudomonas aeruginosa and Acinetobacter). Semin.
Respir. Infect., 15: 287-298.
Chen BM, Xia LW and Zhao RQ (1997). Determination
of NGNG-dimethylarginine in human plasma by high-
performance liquid chromatography. J Chromatogr B
Biomed Sci Appl., 2: 467-471.
Ding Y, Vaziri ND, Coulson R, Kamanna VS and Roh DD
(2000). Effects of simulated hyperglycemia, insulin,
and glucagon on endothelial nitric oxide synthase
expression. Am. J. Physiol. Endocrinol. Metab., 279:
11-17.
Investigation of anti-cancer effects of cherry in vitro
Pak. J. Pharm. Sci., Vol.27, No.3, May 2014, pp.587-592
592
Fu M, Wang C, Li Z, Sakamaki T and Pestell RG (2004).
Minireview: cyclin D1: Normal and abnormal
functions. Endocrinology, 145: 5439-5447.
Galgóczy L, Hevér T, Orosz L, Krisch J, Vágvölgyi C,
Tölgyesi M and Papp T (2009). Growth inhibition
effect of fruit juices and pomace extracts on the enteric
pathogens Campylobacter jejuni and Salmonella ser.
Typhimurium. Internet J. Microbiol., 7(1): 7317.
Harborne JB and Williams CA (2000). Advances in
flavonoid research since 1992. Phytochemistry, 6: 481.
Hebeler BH, Chatterjee AN and Young FE (1973).
Regulation of the bacterial cell wall: Effect of
antibiotics on lipid biosynthesis. Antimicrob. Agents
Chemother., 4: 346-353.
Jafari AA, Hossein S, Karimi F and Pajouhi M (2008).
Effects of sour cherry juice on blood glucose and some
cardiovascular risk factors improvements in diabetic
women. Nutr. Food Sci., 38(4): 355-360.
Jeong SH, Bae IK, Park KO, An YJ, Sohn SG, Jang SJ,
Sung KH, Yang KS, Lee K, Young D and Lee SH
(2006). Outbreaks of imipenem resistant Acinetobacter
baumannii producing carbapenemases in Korea. J
Microbiol, 44: 423-431.
Kamei H, Hashimoto Y, Koide T, Kojima T, Hasegawa M.
(1998). Anti-tumor effect of methanol extracts from red
and white wines. Cancer Biother. Radiopharm., 13:
447-452.
Kang SY, Seeram NP, Nair MG and Bourquin LD (2003).
Tart cherry anthocyanins inhibit tumor development in
ApcMin mice and reduce proliferation of human colon
cancer cells. Cancer Lett., 194: 13-19.
Kim DO, Heo HJ, Kim YJ, Yang HS and Lee CY (2005).
Sweet and Sour Cherry Phenolics and Their Protective
Effects on Neuronal Cells. J. Agric. Food Chem.,
53(26): 9921-9927.
Klevens RM, Morrison MA, Nadle J, Petit S and
Gershman K et al. (2007). Invasive Methicillin-
Resistant Staphylococcus Aureus Infections in the
United States. JAMA, 298:1763-1771.
Krishkov EY (2007). Тrends in World Sour Cherry
Production and State of Sector in Bulgaria after
Transition Period (1997-2006). J. Agric Res., 45(4):
311-318.
Lee YL, Cesario T, Wang Y, Shanbrom E and Thrupp L
(2003). Antibacterial activity of vegetables and juices.
Nutrition, 19: 994-996.
Liu T, Bauskin AR, Zaunders J, Brown DA and Pankurst
S (2003). Macrophage inhibitory cytokine 1 reduces
cell adhesion and induces apoptosis in prostate cancer
cells. Cancer Res., 63: 5034-5040.
Mariette S, Tavaud M, Arunyawat U, Capdeville G,
Millan M and Salin F (2010). Population structure and
genetic bottleneck in sweet cherry estimated with SSRs
and the gametophytic self-incompatibility locus. BMC
Genetics, 11: 77.
Mas T, Susperregui J, Berke B, Cheze C, Moreau S,
Nuhrich A and Vercauteren J (2000). DNA triplex
stabilization property of natural anthocyanins.
Phytochemistry, 53: 679-687.
Mc Carty MF (2004). Vascular endothelium is the organ
chiefly responsible for the catabolism of plasma
asymmetric dimethylarginine an explanation for the
elevation of plasma ADMA in disorders characterized
by endothelial dysfunction. Med. Hypotheses, 63: 699-
708.
Rothfield L and Horecker BL (1964). The role of cell-wall
lipid in the biosynthesis of bacterial
lipopolysaccharide. Proc Natl. Acad. Sci. USA., 52:
939-946.
Sarma AD and Sharma R (1999). Anthocyanin-DNA
copigmentation complex: mutual protection against
oxidative damage. Phytochemistry, 52(7): 1313-1318.
Stockinge EJ, Mulinix CA, Long CM, Brettin TS and
Lezzoni AF (1996). A Linkage Map of Sweet Cherry
Based on RAPD Analysis of a Microspore-Derived
Callus Culture Population. J. Hered 87(3): 214-218.
Turkey Fruit Juice Industry-Statistical Evaluation (2000-
2008).http://www.meyed.org.tr/content/files/istatistikle
r/2008.pdf Accessed date 15.02.2011 (Turkish).
Valkonen VP, Päivä H, Salonen JT, Lakka TA, Lehtimäki
T, Laakso J and Laaksonen R (2001). Risk of acute
coronary events and serum concentration of
asymmetrical dimethylarginine. Lancet, 358: 2127-
2128.
Vallance P, Leone A, Calver A, Collier J and Moncada S
(1992). Accumulation of an endogenous inhibitor of
nitric oxide synthesis in chronic renal failure. Lancet,
339: 572-575.
Vaughan J and Geissler C (2009). The New Oxford Book
of Food Plants Oxford University Press, New York,
p.74.
Wang H, Nair MG, Stasburg GM, Booren AM and Gray JI
(1999). Antioxidant polyphenols from tart cherries
(Prunus cerasus). J. Agric Food Chem., 47: 840-844.
Yamaguchi K, Liggett JL, Kim NC and Baek SJ (2006).
Anti-proliferative effect of horehound leaf and wild
cherry bark extracts on human colorectal cancer cells.
Oncol. Rep., 15(1): 275-281.
... 2=Middle East: includes interior of Asia minor, all of Transcaucasia, Iran, and the highlands of Turkmenistan. Middle East is not only the cradle of civilization for modern humans (Fernandes et al., 2012;Witas et al., 2013) and about 70% of world's petroleum reserves, but also center of origin for 83 species in crop (Vavilov, 1951;Ladizinsky,1998), some functional foods with anticancer including to hawthorn, apple, cherry, barley, oats, and pomegranate Zeng et al., 2013;Ogur et al., 2014); Up to now, Middle East is major production region for these functional crop. The anticancer effects of polyphenolic extract from hawthorn fruit in human MCF-7 breast cancer cells, which is an excellent source of natural chemopreventive agents in the treatment of breast cancer . ...
... Apple oligogalactan could potentiate the growth inhibitory effect of celecoxib on colorectal cancer through influencing the expression and function of cyclooxygenase-2 and phosphorylation of MAPKs, which suggests a new possible combinatorial strategy in colorectal cancer therapy . Cherry is popular preserve in Turkish cuisine, its consumption may decrease the risk of cancer and chronic diseases (Ogur et al., 2014). Protocatechualdehyde from the root of the herb S. miltiorrhiza and barley tea plants possesses antiproliferative and pro-apoptotic properties in human colorectal cancer cells (Choi et al., 2014). ...
... 4=China-Korea Center: is not only the cradle of civilization for modern humans, but also center of origin for 136 species in crop (Vavilov, 1951;Ladizinsky, 1998;Wikipedia, 2013), some functional foods with anticancer including to rice, buckwheat, tea, millet, soybean, adzuki bean,Chinese yam, radish, onion, cucumber, Chinese apple, peach, apricot, walnut, litchi, hemp, cabbage, hullless barley and cherry (Miyoshi et al., 2011;Hsu et al., 2012;Xu et al., 2012;Hattori et al., 2013;Zeng et al., 2013;Benyamina, 2014;Gao et al., 2014;Ghoneum et al., 2014;Hardman, 2014;Ogur et al., 2014;Panda et al., 2014;Shan et al., 2014;Vizzotto et al., 2014;Wang et al., 2014;Hajiaghaalipour et al., 2015). Up to now, China especially Yunnan province is major production region for these functional crop. ...
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Cancer is the leading cause of death around the world. Anticancer activities from many functional food sources have been reported in years, but correlation between cancer prevalence and types of food with anticancer activities from crop origin center in the world as well as food source with human migration are unclear. Hunger from food shortage is the cause of early human evolution from Africa to Asia and later into Eurasia. The richest functional foods are found in crop origin centers, housing about 70% in the world populations. Crop origin centers have lower cancer incidence and mortality in the world, especially Central Asia, Middle East, Southwest China, India and Ethiopia. Asia and Africa with the richest anticancer crops is not only the most important evolution base of humans and origin center of anticancer functional crop, but also is the lowest mortality and incidence of cancers in the world. Cancer prevention of early human migrations was associated with functional foods from crop origin centers, especially Asia with four centers and one subcenter of crop origin, accounting for 58% of the world population. These results reveal that coevolution between human's anticancer activities associated with functional foods for crop origin centers, especially in Asia and Africa.
... The anti-cancer effects of sour cherry have been studied previously, with anthocyanins as their anti-cancer bioactive components [3,29]. Ogur et al. [30] demonstrated the antiproliferative effect of encapsulated cherry ethanol extract in the MCF7 cell line at lower concentrations (50 µg/mL inhibited cell growth by 37%). In an in vitro study, Martin and Wooden also demonstrated that sour cherry juice induced apoptosis in MCF7 cells [29]. ...
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The objective of this work was the valorisation of sour cherry (Prunus cerasus L.) pomace as a source of biologically active compounds. To formulate microcapsules, polyphenolic compounds were extracted and encapsulated with maltodextrin as wall material, by freeze-drying. An in vitro digestion study was carried out on obtained encapsulates but also on sour cherry pomace extract and sour cherry pomace freeze-dried powder. The results indicated that encapsulation, as well as freeze-drying, provided a good protective effect on bioactive compounds during digestion. Furthermore, the potential antiproliferative and cytotoxic activities of encapsulates and sour cherry pomace extract were evaluated using breast adenocarcinoma MCF7 cell lines, colon adenocarcinoma HT-29 cell lines, and noncancer cell line. Encapsulates and sour cherry pomace extract showed variable anti-proliferative activity towards all cell lines. Obtained results showed that encapsulation of sour cherry pomace could be useful for improving the stability of polyphenolic compounds in the gastrointestinal tract. The results highlight the bioactive potential of sour cherry pomace as a nutraceutical resource and the protective effects of microencapsulation on the digestion of bioactive compounds.
... Inhibition was achieved by feeding mices with 0.8-1.6 mg/day, this effect was mediated by inhibition of metalloproteinases gene expression [118]. Sour cherry extracts were also reported with antiproliferative activity and induction of apoptosis in mammary adenocarcinoma (MCF-7) and mouse mammary tumour cell (4T1) breast cancer cells lines [119]. Moreover, peach and plum extracts and purified phenolic compounds can inhibit growth and induce differentiation of colon cancer cells [120], and plums extracts have similar beneficial effect in vivo [121]. ...
... The cherry extract containing anthocyanins inhibited the proliferation of human colon cancer cells HCT 116 and HT 29 (Kang et al. 2003). Bobe et al. (2006) demonstrated that the cherry extract in combination with sulindac reduced the incident of tumorigenesis in small intestine and also inhibited the proliferative activities in human (MCF-7) and mouse (4 T1) breast cancer cells (Ogur et al. 2014). ...
Chapter
Fruits of the family, Rosaceae (Apple, cherry, Peach, strawberry, rose, raspberry) are rich source of phenolic and antioxidant compounds having anticancer properties. The present chapter discusses the detail information about anticancer compounds of strawberry, raspberry, peach, apple, cherry and rose and also the genes responsible for the biosynthesis, accumulation and transport of anticancer compounds during growth and maturation of fruits. The transcriptome expression was performed to find putative genes responsible for anticancer compounds during the biosynthesis and transporter genes. It is revealed form the promoter analysis that cis-acting element is responsible for the regulation of anticancer compounds. Thus, CRISPR/Cas9 enhanced the biosynthesis of anticancer compounds during fruit development and maturation stages. CRISPR/Cas9 will be used for the silencing of genes which putatively inhibit the formation of anti-cancer compounds and also up-regulate biosynthesis and transporter genes mediated by CRISPR/Cas9to enhance their accumulation in these fruits.
... Inhibition was achieved by feeding mices with 0.8-1.6 mg/day, this effect was mediated by inhibition of metalloproteinases gene expression [118]. Sour cherry extracts were also reported with antiproliferative activity and induction of apoptosis in mammary adenocarcinoma (MCF-7) and mouse mammary tumour cell (4T1) breast cancer cells lines [119]. Moreover, peach and plum extracts and purified phenolic compounds can inhibit growth and induce differentiation of colon cancer cells [120], and plums extracts have similar beneficial effect in vivo [121]. ...
Chapter
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Fruits constitute one of the most important sources of phytochemicals in human diet. Stone fruits, such as peaches, plums, almonds, apricots and cherries have been investigated concerning their therapeutic effects in the prevention of a range of diseases. The consumption of these fruits is related with the lower prevalence of diabetes, overweight or general obesity, lower risk for estrogen receptor-negative tumors and cardiovascular protection among others. Phenolic compounds, predominantly flavonoids and phenolic acids, are the main phytochemicals in stone fruits. Considering the importance of stone fruits as a source of biologically active compounds the present chapter aims to provide the current findings in this field and the main implications to human health associated with its consumption.
... The prevalence rate of CAM utilization among use has made significant contributions in the healthcare management [3], the recent growth in the prevalence rate of CAM use among patients have raised concerns about the toxicity and the potential herbal-drug, herbal-disease, and herbal-herbal interactions [6,7]. However, some of the therapeutic potentials of some herbs have been established in vitro [8]. ...
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Many studies have been conducted in health-care settings with regards to complementary and alternative medicine (CAM) use among patients. However, information regarding CAM use among patients in the emergency department (ED) is scarce. The aim of this article was to conduct a systematic review of published studies with regards to CAM use among the ED patients. A literature search of published studies from inception to September 2015 was conducted using PubMed, Scopus, and manual search of the reference list. 18 studies that met the inclusion criteria were reviewed. The prevalence rate of CAM use among ED patients across the studies ranged of 1.4-68.1%. Herbal therapy was the sub-modality of CAM most commonly used and frequently implicated in CAM-related ED visits. Higher education, age, female gender, religious affiliation, and chronic diseases were the most frequent factors associated with CAM use among the ED patients. Over 80% of the ED physicians did not ask the patients about the CAM therapy. Similarly, 80% of the ED patients were ready to disclose CAM therapy to the ED physician. The prevalence rate of CAM use among patients at ED is high and is growing with the current increasing popularity, and it has been a reason for some of the ED visits. There is a need for the health-care professionals to receive training and always ask patients about CAM therapy to enable them provide appropriate medical care and prevent CAM-related adverse events.
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Inflammation is an underlying cause of or a contributing factor to a number of chronic conditions, including hypertension, insulin resistance, arthritis, and cognitive disorders. A chronic inflammatory state is also associated with aging. Tart cherry (TC) has been extensively studied for its ability to prevent or treat inflammatory diseases and their associated risk factors. TC contains active compounds, including polyphenols that may contribute to its antioxidant and anti-inflammatory effects. Inflammatory signaling pathways regulate the recruitment of inflammatory cells important for the pathogenesis of disease. Whole TC, individual compounds, and their metabolites may be viable treatment options because they can target molecules involved in inflammatory pathways. In this review, the effectiveness of TC in reducing inflammatory markers associated with chronic diseases and the effects of the active compounds in TC and their metabolites on inflammatory pathways are discussed. The main polyphenols present in TC include cyanidins, kaempferol, quercetin, melatonin, neochlorogenic acid, chlorogenic acid, and 3-coumaroylquinic acid. Evidence supports an association between TC intake and reduced risk for inflammatory disease, which may be due to the effects of active compounds in TC on inflammatory pathways, such as NF-κB and Mitogen-activated protein kinase.
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The present work evaluated the anticancer properties of sweet cherry (Prunus avium) extract on human prostate cells. Several sweet cherry cultivars from Fundão (Portugal) were methanol-extracted and their phytochemical composition characterized. The Saco “late harvest” extract was highly-enriched in anthocyanins and selected for use in biological assays. Non-neoplastic (PNT1A) and neoplastic (LNCaP and PC3) human prostate cells were treated with 0–2,000 μg/ml of extract for 48–96 h. Cell viability was evaluated by the MTT assay. Apoptosis, oxidative stress, and glycolytic metabolism were assessed by Western blotting and enzymatic assays. Glucose consumption and lactate production were measured spectrophotometrically. Saco cherry extract diminished the viability of neoplastic and non-neoplastic cells, whereas enhancing apoptosis in LNCaP. Cherry extract-treatment also diminished oxidative damage and suppressed glycolytic metabolism in LNCaP cells. These findings widened the knowledge on the mechanisms by which cherry extract modulate cell physiology, demonstrating their broad action over the hallmarks of cancer.
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Crude methanol extracts of red and white wines were added to diethyl ether in order to divide them into the anthocyanin fraction (insoluble in diethyl ether) and fractions containing other flavonoids and their derivatives (soluble in diethyl ether). However, the white wine did not contain anthocyanins (all of the methanol extract was soluble in diethyl ether). When HCT-15 cells, derived from human colon cancer or AGS cells, derived from human gastric cancer, were cultured with these fractions, the anthocyanin fraction from the red wine and the non-anthocyanic substances extracted from red and white wines suppressed the growth of the cells, and the suppression rate by the anthocyanin fraction was significantly higher than that of the other fractions. Thin-layer chromatographic analysis revealed mostly delphinidin in the anthocyanin fraction. The other fractions contained mostly flavonoids and their derivatives. The sugars in all fractions were mainly glucose, fucose, and fructose. Flow cytometric study suggested that the anthocyanin fraction blocked mostly S, G2, and M phase, and the non-anthocyanic flavonoids also blocked these phases, although the histographic pattern varied depending an the fractions. Methanol insoluble but wafer soluble fractions (mostly free sugars) of red and white wines did not show such suppressive effects.
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Purpose – Some studies on anthocyanins have revealed their antioxidant activity and beneficial effects for diabetes control and reducing the risk of coronary heart diseases. It has been found that sour cherries contain high levels of anthocyanins that possess insulin‐releasing stimulatory properties on pancreatic β‐cells in vitro. The purpose of this paper is to investigate whether concentrated sour cherry juice (CSCJ) beneficially alters serum glucose and some cardiovascular risk factors in diabetes type 2 subjects. Design/methodology/approach – In this quasi‐experimental study, 19 diabetic women with FBS ≥ 110 mg/dl were recruited from patients referred to the Diabetes Clinic of Shariati Hospital. Subjects were asked to consume 40 g of CSCJ daily for 6 weeks. Before the onset of the study (week 0) and after 6 weeks, weight and blood pressure measurements were done and fasting blood samples were drawn. FBS, hemoglobin A1c (HbA1c) and blood lipid profiles were measured. In addition, a 24‐hour food record was taken from all of the individuals in both stages. The Wilcoxon signed test was used for statistical analysis. Findings – After six weeks' consumption of CSCJ, significant reductions in body weight (p < 0.01), blood pressure and HbA1c (p < 0.05) was seen. Total cholesterol and LDL‐C decreased significantly in a sub‐group of patients (n = 12) with LDL‐C ≥ 100 mg/dl as well. Originality/value – Based on the results of this study, consuming 40 g/day of CSCJ decreases body weight, blood pressure and HbA1c in diabetes type 2 women after 6 weeks and improves blood lipids in diabetic patients with hyperlipidemia.
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The antibacterial effect of fruit juices and pomace extracts from 13 wild and cultivated fruits (Prunus avium, P. cerasus, P. armeniaca, Crataegus monogyna, Morus alba, M. nigra, Ribes nigrum, R. rubrum, R. uva-crispa, R. nidigrolaria, Rubus idaeus, R. fruticosus) against two foodborne enteric pathogens (Salmonella ser. Typhimurium, Campylobacter jejuni) was evaluated by broth microdilution assays. Juices and/or extracts of sour cherry, apricot, raspberry, blackcurrant, redcurrant, gooseberry and jostaberry efficiently inhibited the growth of both bacteria (growth ≤ 25%). Juices and extracts from cherry (red and yellow cultivars), hawthorn, blackberry and pomace extracts from black and white mulberry had a similar strong inhibitory effect on the growth of C. jejuni, but had weak or no effect on S. ser. Typhimurium. Sour cherry, jostaberry and raspberry pomace extracts revealed a substantial antibacterial effect at both acidic and neutral pH. - See more at: http://www.ispub.com/journal/the-internet-journal-of-microbiology/volume-7-number-1/growth-inhibition-effect-of-fruit-juices-and-pomace-extracts-on-the-enteric-pathogens-campylobacter-jejuni-and-salmonella-ser-typhimurium.
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The DNA triplex stabilization property of seven natural anthocyanins (five monoglucosides and two diglucosides) has been measured by the mean of triplex thermal denaturation experiments. We have noticed a difference between the diglucosides that do not modify this melting temperature and the monoglucosides (namely 3-O-β-D-glucopyranoside of malvidin, peonidin, delphinidin, petunidin and cyanidin) which present a weak but significant stabilizing effect. It appears clearly that the difference between the two series could be due to the supplementary sugar moiety at the 5 position for the diglucosylated compounds, that would make them too crowded to allow interaction with the triplex. Among the monoglucoside series, the most active compounds are the only ones to embody a catechol B-ring in their structure that could be important for such an interaction. The need to have pure and fully characterized compounds to run these measurements, made it possible for us to unambiguously assign the 1H and 13C NMR spectra with the help of 2D NMR experiments. Thus, missing data of compounds not totally described earlier, are provided herein.
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The absorption spectra of a cyanidin derivative showed a 15–20 nm bathochromic shift when mixed with calf thymus DNA (ctDNA), indicating formation of a cyanidin-DNA copigmentation complex. Exposure of either cyanidin or ctDNA individually to hydroxyl radicals (OH) obtained in the Fenton reaction between ferrous ions and hydrogen peroxide caused severe oxidative damage. However, formation of cyanidin-DNA complex prior to exposure to OH• protected both the cyanidin and ctDNA from the oxidative damage. These results suggest that cyanidin-DNA copigmentation might be a possible defense mechanism against oxidative damage of DNA and may have in vivo physiological function attributable to the antioxidant ability of anthocyanins.
NG,NG-Dimethylarginine (asymmetric dimethylarginine, ADMA) can be directly separated and measured from deproteinized human plasma using o-phthaldialdehyde-mercaptoethanol (OPA reagent) as a fluorogenic reagent by reversed-phase high-performance liquid chromatography. The mean recovery of ADMA was over 96% and the inter- and intra-assay coefficients of variation of amounts were lower than 3.80% and those of retention time were below 0.37% for five runs. The detection limit of the assay is 1 pmol when the signal-to-noise is 3:1. It was observed that the concentration of ADMA was significantly elevated in plasma of patients with pregnancy induced hypertension (PIH) in contrast to healthy pregnant women.