Abstract and Figures

Orange juice (OJ) is among the most consumed fruit juices worldwide, and its chemopreventive action is fairly addressed in the literature. This review critically presents the available evidence linking OJ with cancer chemoprevention and on discussing the putative mechanisms and negative health effects. The chemopreventive action of OJ is related to its effect on metabolic enzymes and its antiinflammatory, cytoprotective/apoptotic, hormonal, cell signaling-modulating, antioxidant, and antigenotoxic effects. Most studies on OJ are in vitro, and few are conducted in vivo. Results from in vitro studies must be interpreted carefully because these findings do not consider in vivo bioavailability. However, such results are useful for studying the impact of different processing and storage methods on OJ's chemopreventive effect. Evidence of OJ's chemoprevention in humans is limited. OJ is antimutagenic in bacteria and antigenotoxic in humans and rodents. Studies using rodent cancer models showed that OJ is cancer chemopreventive, influencing either the induction stage or the promotion stage. The composition and, therefore, the chemopreventive action of OJ might be influenced by different cultivars, climates, extraction methods, packaging, storage temperatures, and shelf lives, among other factors. Epidemiological studies and randomized controlled intervention studies in humans evaluating the chemopreventive effect of OJ, taking into consideration variability in OJ composition, are needed.
Content may be subject to copyright.
This article was downloaded by: [UNISC]
On: 06 August 2013, At: 15:03
Publisher: Routledge
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House,
37-41 Mortimer Street, London W1T 3JH, UK
Nutrition and Cancer
Publication details, including instructions for authors and subscription information:
http://www.tandfonline.com/loi/hnuc20
Orange Juice and Cancer Chemoprevention
Silvia Isabel Rech Franke
a
b
, Temenouga Nikolova Guecheva
b
, João Antonio Pêgas
Henriques
c
b
& Daniel P
a
b
a
PPG em Promoção da Saúde, Universidade de Santa Cruz do Sul , Santa Cruz do Sul , RS ,
Brasil
b
Genotox/PPGBCM/PPGBM, Universidade Federal do Rio Grande do Sul , Porto Alegre , RS ,
Brasil
c
Instituto de Biotecnologia , Universidade de Caxias do Sul , Caxias do Sul , RS , Brasil
Published online: 06 Aug 2013.
To cite this article: Nutrition and Cancer (2013): Orange Juice and Cancer Chemoprevention, Nutrition and Cancer, DOI:
10.1080/01635581.2013.817594
To link to this article: http://dx.doi.org/10.1080/01635581.2013.817594
PLEASE SCROLL DOWN FOR ARTICLE
Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained
in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no
representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the
Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and
are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and
should be independently verified with primary sources of information. Taylor and Francis shall not be liable for
any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever
or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of
the Content.
This article may be used for research, teaching, and private study purposes. Any substantial or systematic
reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any
form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://
www.tandfonline.com/page/terms-and-conditions
Nutrition and Cancer, 1–11, 2013
Copyright
C
Taylor & Francis Group, LLC
ISSN: 0163-5581 print / 1532-7914 online
DOI: 10.1080/01635581.2013.817594
Orange Juice and Cancer Chemoprevention
Silvia Isabel Rech Franke
PPG em Promoc¸
˜
ao da Sa
´
ude, Universidade de Santa Cruz do Sul, Santa Cruz do Sul, RS, Brasil,
and Genotox/PPGBCM/PPGBM, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
Temenouga Nikolova Guecheva
Genotox/PPGBCM/PPGBM, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
Jo
˜
ao Antonio P
ˆ
egas Henriques
Genotox/PPGBCM/PPGBM, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil,
and Instituto de Biotecnologia, Universidade de Caxias do Sul, Caxias do Sul, RS, Brasil
Daniel Pr
´
a
PPG em Promoc¸
˜
ao da Sa
´
ude, Universidade de Santa Cruz do Sul, Santa Cruz do Sul, RS, Brasil,
and Genotox/PPGBCM/PPGBM, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
Orange juice (OJ) is among the most consumed fruit juices
worldwide, and its chemopreventive action is fairly addressed in
the literature. This review critically presents the available evidence
linking OJ with cancer chemoprevention and on discussing the
putative mechanisms and negative health effects. The chemopre-
ventive action of OJ is related to its effect on metabolic enzymes
and its antiinflammatory, cytoprotective/apoptotic, hormonal, cell
signaling-modulating, antioxidant, and antigenotoxic effects. Most
studies on OJ are in vitro, and few are conducted in vivo. Re-
sults from in vitro studies must be interpreted carefully because
these findings do not consider in vivo bioavailability. However, such
results are useful for studying the impact of different processing
and storage methods on OJ’s chemopreventive effect. Evidence of
OJ’s chemoprevention in humans is limited. OJ is antimutagenic
in bacteria and antigenotoxic in humans and rodents. Studies us-
ing rodent cancer models showed that OJ is cancer chemopreven-
tive, influencing either the induction stage or the promotion stage.
The composition and, therefore, the chemopreventive action of
OJ might be influenced by different cultivars, climates, extraction
methods, packaging, storage temperatures, and shelf lives, among
other factors. Epidemiological studies and randomized controlled
intervention studies in humans evaluating the chemopreventive ef-
fect of OJ, taking into consideration variability in OJ composition,
are needed.
Submitted 31 August 2012; accepted in final form 28 May 2013.
Address correspondence to Daniel Pr
´
a, PPG em Promoc¸
˜
ao da
Sa
´
ude, Universidade de Santa Cruz do Sul, UNISC. Av. Independ
ˆ
encia,
2293, Pr
´
edio 42, sala 4206. CEP 96815-900 Santa Cruz do Sul-RS,
Brasil. Fax: +5551 3717-1855. E-mail: daniel
pra@yahoo.com
REVIEW SCOPE
In this review, we will present the available evidence of the
role of orange juice (OJ) in cancer chemoprevention discussing
putative mechanisms involved in this process. Later we will dis-
cuss the potential toxicity of OJ and finally, we will critically
discuss the available data in terms of evidence-based medicine.
Primary studies were retrieved from Pubmed and Web of Sci-
ence with combinations of the terms: orange juice and oxida-
tive stress, antioxidant, genotoxic, mutagenic, DNA damage,
cancer, carcinogenic, anticarcinogenic, anti-inflammatory, im-
mune-modulatory, and cell signaling. Studies were manually
selected to be included in the review. Additional studies were
selected as references from the cited papers. Only studies using
commercial or OJ prepared by the researchers were included in
this review. Studies based on extracts from oranges were avoided
and only mentioned to corroborate primary evidence provided
by results with OJ.
OJ
Oranges are among the most consumed fruits worldwide,
which reflects the fact they have been cultivated since ancient
times. They are widely grown in warm climates worldwide;
Brazil and the United States produce about 50% of the total
world supply. The flavors of oranges vary from sweet to sour.
The fruit is commonly peeled and eaten fresh or squeezed for its
juice. There are 2 kinds of orange; Citrus sinensis is called sweet
orange and Citrus aurantium is called sour orange (other names
include bitter orange, bigarade orange, and Seville orange)(1).
Sweet oranges can be further divided into 2 varieties, blond and
blood oranges, based in their pulp coloration. Blond oranges
1
Downloaded by [UNISC] at 15:03 06 August 2013
2 S. I. R. FRANKE ET AL.
have a smooth peel, soft pulp, weakly winged leafstalk (2) and
regular-colored juice. Blood (red pulp) oranges are richer in
anthocyanins and produce darker juices (3).
OJ is among the most consumed fruit juices and OJ itself and
its constituents may exert various biological effects (4–7). OJ
is considered among the top micronutrient-dense juices (i.e., a
food that provides substantial amounts of vitamins and miner-
als and relatively fewer calories) (8). Recent evidence from a
study comparing adults over 19 years of age (n = 8,861), who
were consumers and nonconsumers of OJ participating in the
National Health and Nutrition Examination Survey, 2003–2006
have shown that consumption of OJ (usual per capita intake
of 100% OJ was 50.3 mL/day) was associated with better diet
quality and an increased prevalence of meeting the estimated
average requirement for key nutrients (mainly vitamin C and
folate) as well as other biomarkers of positive health outcomes,
including lower total cholesterol and LDL levels. The study also
indicated that consumers of OJ had lower mean body mass index
and a decreased risk of obesity (9).
OJ can provide substantial amounts of vitamin C and folic
acid, 2 well-known substances that can reduce DNA damage
levels and, thus, the risk of cancer (10, 11). OJ also pro-
vides substantial amounts of phenolic substances, particularly of
flavonoids, that can also have chemopreventive action (12). The
effect of these substances will be addressed later in this review.
The composition of oranges and, therefore OJ, varies de-
pending on varietal, climatic conditions, soil composition, light
and pathogen exposure, and maturation stage during harvesting
period, as well as on storage and post-harvest processing (13).
Thus, it is important to highlight that the reviewed evidence
will depend on the experimental designs, the purpose of the
study, the doses of OJ administered, the treatment scheme, as
well as the system or subsystem (e.g., cell culture versus hu-
man populations) under study; therefore indicating extensive
differences among the studies.
OJ AND CANCER CHEMOPREVENTION
Table 1 summarizes epidemiological and murine cancer-
model studies with OJ. Primary epidemiological evidence is
inexistent for OJ. The few studies available indicated that OJ
was not associated with a reduced risk of squamous cell carci-
noma of the skin (14) and there is evidence that it might increase
the risk of melanoma (15). Evidence also indicates OJ can re-
duce the risk of leukemia in children (16).
Conversely, the murine models of cancer (Table 1, bottom)
have shown that OJ can be chemoprotective against mammary
(17), hepatic (18), and colon cancer (19). For mammary car-
cinogenesis, female Sprague-Dawley were treated with 5 mg
dimethylbenz[a]anthracene, as a single intragastrically dose at
approximately 50 days of age (while in diestrus), and double-
strength OJ was administered ad libitum (108.5 mL per rat per
day) for the next 15 wk. OJ was able to reduce the mammary
tumor weight and tumor burden (17). This effect seemed to be
mediated by the synergistic action of hesperetin with other com-
pounds that have anticancer activity (20). In hepatic carcinogen-
esis chemoprotection, Fischer 344 male rats were treated daily
with 250 μg/kg aflatoxin B-1 after reaching 85 g, with 10 al-
ternate doses, and with OJ extract (0.5 mg/kg) either concomi-
tantly with aflatoxin B-1 treatment (induction) or every 2 days
after the induction for 12 wk (promotion). OJ extract substan-
tially decreased the number of gamma-glutamyl transpeptidase-
positive foci in liver when administered during the initiation
period, whereas during the promotion period caused a decrease
in the average diameter of the foci. The total volume of foci was
markedly reduced by OJ during either period (18). Therefore,
OJ extract treatment in this murine models of cancer seemed to
inhibit the biochemical and cellular events associated to either
initiation or promotion (18). In colon carcinogenesis chemopre-
vention experiments, male Fischer 344 rats received 15 mg/kg
azoxymethane by subcutaneous injections at 22 and 29 days of
age and 1 wk later pasteurized OJ in place of drinking water for
another 28 wk. At the end of the experiments, colon tumor in-
cidence and proliferating cell nuclear antigen-positive nuclei in
colon crypts was reduced, and there was a strong trend toward
a smaller average tumor burden (mg tumor/rat) for the group
drinking OJ (19). This results showed an inhibition of clonal
expansion of transformed cells into visible polyps or tumors
when drinking water was replaced by pasteurized OJ (19).
CHEMOPROTECTIVE MECHANISMS OF OJ
Figure 1 summarizes the chemopreventive mechanisms
of OJ, including antioxidant, antigenotoxic, cytoprotec-
tive/apoptotic, antiinflammatory, cell-signaling, phytoestrogen,
antimicrobial, and antiviral effects as well as an effect on
metabolism and excretion of xenobiotics. These effects are de-
tailed in the following sections.
Antioxidant Potential of OJ
There are many studies evaluating the antioxidant potential
of OJ, mostly in vitro. All in vitro studies have shown that
OJ has considerable antioxidant potential. The high content of
flavonones is linked to OJ’s antioxidant potential. Juices rich in
flavonones are the second-best antioxidant, after fruits rich in
anthocyanins (red, purple, or blue fruits) (21–23).
Certain factors affect in vitro antioxidant potential. High tem-
peratures during thermal treatment (3, 24, 25) or during storage
(>20
C) and long-term storage (>4 mo) (4, 25–27) decrease
antioxidant activity due to phytochemical composition changes
(3, 27, 28). Sweetening seems to lead to the extinction of most
antioxidant activity (4).
To date, it is not clear how much each OJ constituent con-
tributes to the overall antioxidant activity. It is possible that
some compound act at short-term losing their antioxidant ca-
pacity (e.g., vitamin C) and other such as phenolics might retain
their antioxidant potential for longer periods (e.g., phenols are
stable for long periods but change in term of their relative com-
ponents) (29, 30). The test system used to evaluate it, which
Downloaded by [UNISC] at 15:03 06 August 2013
ORANGE JUICE AND CANCER 3
TABLE 1
Summary of the epidemiological and murine models studies evaluating orange juice (OJ) chemoprevention
Food Type of study/endpoint Result Reference
Epidemiological studies
OJ, oranges, or citrus peel Population-based case-control study
evaluating the relationship between
citrus consumption and the history of
squamous cell carcinoma of the skin in
an elder Southwestern U.S. population
(n = 242 cases and 228 controls)
Consumption of OJ [OR = 1.00
(0.83–1.20)] or [OR = 0.89
(0.71–1.11)] were not associated to the
reduction of skin carcinoma risk, but
citrus peel intake reduced its risk [OR =
0.66 (0.45–0.95)].
(14)
OJ Follow-up study in 2 Nurse’s Health Study
cohorts evaluating the relationship
between vitamins and certain foods and
the risk of melanoma (n = 162,000)
The frequency of OJ consumption was
linearly associated with increasing
relative risk of melanoma (P < 0.01)
(15)
OJ Case-control study aiming to study the
relation between child’s early diet and
risk of childhood leukemia in diverse
California population (n = 328
case-control sets)
OJ regular consumption during the first
2 years of life was associated with a
reduction in risk of childhood leukemia
diagnosed between the ages of 2 and
14 yr [OR = 0.54 (0.31, 0.94)]
(16)
Murine models studies
Double-strength OJ Evaluation of mammary tumors induced
by 7,12-dimethylbenz[a]anthracene in
female Sprague-Dawley rats
Mammary tumor development was
delayed and tumor burden (grams of
tumor/rat) was reduced
(17, 20)
OJ
a
Evaluation of preneoplastic foci in the
liver of Fischer 344 male rats treated
with aflatoxin B-1
Decrease in number (initiation) and
volume (promotion) of gamma-glutamyl
transpeptidase-positive foci in liver
(18)
Pasteurized OJ in place of
drinking water
Evaluation of colon cancer in male Fischer
344 rats treated with azoxymethane
Colon tumor incidence was reduced by
22%
(19)
OR = odds ratio with 95% confidence interval (in parenthesis).
a
An extract of orange juice (OJ) was used to treat the rats.
might help in interpreting their real beneficial effect in vivo, can
also influence the results. Results from in vitro studies must be
interpreted carefully because these findings do not consider in
vivo bioavailability.
Most studies evaluating the antioxidant effects of OJ in hu-
mans and rats showed antioxidant effects. A single intake of
OJ can increase the blood radical scavenging capacity from
90 min (150 mL) (31) to 1 day (300 mL) (32). A single in-
take of OJ (600 mL) also increases the antioxidant capacity for
morethan3h(33).Two-weektreatmentsofhealthy volunteers
(up to 500 mL OJ daily) reduced the levels of lipid peroxi-
dation (34). Dosages equal or greater than 600 mL daily for
more than 2 wk have been shown to induce not only a short
lipid peroxidation decrease but a reduction in DNA oxidation as
well (6, 35). Processing and storage can have little or no influ-
ence or completely abolish antioxidant effects (32), but further
studies are still needed to better understand this issue. In vivo
data also confirm the observation that storage reduces the an-
tioxidant effect of OJ, particularly due to the loss of vitamin C
(32). Results of OJ administration in rats confirmed the effect in
humans, indicating that longer administrations (6 mo) of daily
doses comparable to humans (1 mL for a rat equals 600 mL for
a human) might increase antioxidant capacity in the long term
(36). It is important to mention that the extrapolation of murine
data on OJ administration to effects in humans must be done
with care. Although rodents generally have extensive biological
homology to humans, their ability to synthesize vitamin C (37)
makes them less favorable models for OJ testing. There is a
lack of studies evaluating the impact of factors that impair the
in vitro antioxidant effect with the in vivo antioxidant effect.
Antimutagenic and Antigenotoxic Effects of OJ In Vivo
OJ reduced the mutagenicity induced by different mutagens
as evaluated by the Salmonella/microsome assay (Ames test)
with and without metabolic activation. OJ was more effective as
cotreatment and was most potent inhibitor in relation to other
substances tested (38–40).
Riso et al. (35) showed that lymphocytes of individuals sup-
plemented with blood OJ (600 mL × 21 days) had increased
resistance to H
2
O
2
-mediated DNA damage, as evaluated by
Downloaded by [UNISC] at 15:03 06 August 2013
4 S. I. R. FRANKE ET AL.
FIG. 1. Activities and effects of orange juice on the multistage process of carcinogenesis.
the comet assay. In mice, our data indicate that OJ (a single
dose equivalent to 600 mL in humans) reduced the genotoxi-
city induced by alkylating agents, iron, and copper, also using
the comet assay. Differences were observed depending on the
treatment schedule (OJ as pre- or posttreatment). OJ, tested ei-
ther before or after treatment with the alkylating agent methyl
methanesulfonate, reduced the DNA damage (by approximately
70% as a pretreatment and by approximately 40% as a posttreat-
ment). The pretreatment reduced the genotoxicity of either iron
or copper by approximately 50%. No significant effect was seen
during posttreatment with OJ after treatment with iron and cop-
per (41, 42). Guarnieri et al. (43) evaluated the effect of the
intake of a single portion of blood OJ (300 mL) on mononuclear
blood cell resistance to H
2
O
2
-induced DNA damage (43) by the
comet assay.
The comet assay detects DNA single-strand breaks, alkali-
labile sites, DNA-DNA/DNA-protein crosslinking, and single-
strand breaks associated with incomplete excision repair sites
(44). OJ could therefore reduce these types of DNA damage.
In support of an antimutagenic effect of OJ, a reduction in the
frequency of micronuclei was also detected in mice exposed to
radiation or cyclophosphamide and treated with orange extracts
(45, 46). Oshawa et al. (47) found no suppression of primary
DNA damage in the liver or stomach of mice orally treated
with OJ just before a simultaneous oral dose of morpholine and
NaNO
2
. Such results indicate the DNA-protective effects of OJ
might depend of the nature of the DNA damage generated.
Cytoprotective/Apoptotic, Antiinflammatory, and Cell
Signaling Effects of OJ
There is extensive data supporting the role of inflammation
in cancer, because many cancers arise from sites of infection,
chronic irritation, and inflammation (48). The antiinflammatory
action of plant phytochemicals has been advocated as an impor-
tant mechanism of reducing cancer risk (12). In spite of many
studies addressing the effect of OJ constituents individually,
fewer studies address the effect of the whole OJ in tissues. OJ
significantly reduced plasma concentrations of F2-isoprostanes
Downloaded by [UNISC] at 15:03 06 August 2013
ORANGE JUICE AND CANCER 5
and might also reduce C-reactive protein (34, 49) and markers of
oxidative stress and inflammation, which are typically increased
in the plasma in prooxidative states such as diabetes (33).
In vitro, OJ has differential effects on cell survival. OJ
was shown to increase metabolism and proliferation of cells.
Ekmekcioglu et al. (50) evaluated the effect of OJ on the in-
testinal cell line Caco-2 in terms of toxicity, growth and differ-
entiation. Cells exposed to fresh OJ (50% v/v) exhibited higher
tetrazolium reduction rates in the MTT assay (121.3% of con-
trol). These cells also showed higher succinate-cytochrome c
reductase activities than the other samples, implying that the
contents of fresh OJ, such as ascorbic acid, stimulated mito-
chondrial metabolism (50). Lim and Lim (51) also found 10%
and 30% but not 50% (v/v) OJ could increase Caco-2 cell via-
bility by up to 50% in relation to controls. On the other hand,
extracts from blood OJ were shown to inhibit proliferation of
lung fibroblasts and epithelial prostate cancer (52).
There are, to date, no studies on the effects of OJ on cell
signaling in cancer chemoprevention, but evidence of a cell-
signaling role for individual OJ constituents in cancer chemo-
prevention is accumulating. For instance, both hesperidin (53)
and vitamin C (54) have been shown to induce apoptosis, pos-
sibly by different mechanisms. On the other hand, there is also
growing evidence that vitamin C (100 μM) protects skin cells by
promoting fibroblast proliferation, migration, and replication-
associated base-excision repair of potentially mutagenic DNA
lesions (55). Other compounds such as limonoids are also potent
antiproliferative agents in different cancer cells (56)
Phytoestrogen Effect of OJ
Numerous epidemiological studies suggest that diets rich in
phytoestrogens may be associated with low risk of some can-
cers, especially steroid hormone-dependent cancers (e.g., breast
and prostate) (57). OJ contains low to moderate amounts of
phytoestrogens (58), and hesperidin, one of the major bioac-
tive components of OJ, has been prescribed for the treatment
of hot flashes associated with menopause (59). Naringenin has
also been shown to significantly increase uterine weight in rats
through a tissue-specific effect on estrogen receptor-α distribu-
tion. Possibly because of its flavonoids, OJ can also improve
bone health in orchidectomized old rats with osteoporosis, also
though a hormone-linked pathway (60).
Antimicrobial and Antiviral Effects of OJ
There is growing evidence of the impact of viral and bacte-
rial infections on cancer risk (61). Although there is evidence
of the antimicrobial and antiviral effects of OJ, these aspects
are less studied. It is generally believed that OJ can prevent
common colds, although the only available robust evidence is
that OJ (62) or vitamin C (63) can reduce the severity and dura-
tion of common cold-associated symptoms. Moreover, OJ could
provide a physiological level of ascorbic acid, similar to those
levels achieved by taking high doses of ascorbic acid (62). OJ,
similar to other juices, has been shown to inhibit certain types
of infections. The inhibition of bacterial adherence to bladder
cells has been assumed to account for beneficial effects on the
prevention of urinary tract infections. Zafriri et al. (64) showed
that commercial OJ inhibits the adherence of Escherichia coli
to eukaryotic cells.
Effects of OJ on Bioavailability of Nutrients, Medicines,
and Xenobiotics
OJ has been shown to enhance the absorption of minerals
such as iron (65), aluminum (66), calcium (67), zinc (68), and
selenium (69). OJ was also shown to modulate the absorption
of certain amino acids. There is evidence that OJ increases the
absorption of glycine but not lysine and methionine (70).
OJ has also been shown to interfere with the bioavailability
of several medicines. OJ was shown to reduce the absorption of
acetylsalicylic acid (71), β-adrenergic blocking agents (72, 73),
antihistamines (74), HIV-protease inhibitors (75), chemothera-
peutic agents (76), and hypoglycemic drugs (77). Because of the
potential influence of OJ on drug absorption and metabolism, it
is recommended that patients avoid consuming OJ while taking
medication and that healthcare providers advise against OJ in-
take until any interactions with specific drugs can be clarified in
clinical studies (78).
OJ might influence the bioavailability of substances by dif-
ferent mechanisms, including pH changes in the intestinal lu-
men, chelation, and interference with phase I and II enzymes.
OJ’s acidity, influence on gastric emptying (71), influence on
metabolic enzymes (72), and/or influence on the influx or efflux
of compounds from the interior of cells to the extracellular space
(74, 76, 79) can either stimulate or reduce drug absorption. OJ
can therefore affect the bioavailability of drugs, bioactive food
ingredients, and/or foodborne toxic compounds upon oral up-
take (79).
Putative Compounds in OJ Cancer Chemoprevention
The subject of which component of OJ is more effective in
cancer chemoprevention is controversial (20, 80). Of these, the
flavonoids are the most studied. The antiproliferative effects of
OJ flavonoids are possibly due to regulation of the cell cycle
(17, 20, 56), among other effects. Flavonoids have been shown
to regulate the downstream genes that are responsive to the nu-
clear factor of kappa light-chain enhancer of activated B cells
(NFκB) and mitogen-activated protein kinase (MAPK) signal-
ing cascades. NFκB involves a transcription control mechanism
of cellular responses to stimuli such as oxidative stress, inflam-
mation and bacterial and viral infections that are associated
with cancer (81). MAPK is an enzymatic system that responds
to extracellular mitogenic stimuli and regulates various cellular
activities, such as cell division and proliferation, differentiation
and apoptosis (82, 83).
Heperidin and narigin and their aglycones, hesperitin and
naringinin, respectively, are the most abundant and studies
flavonoids of OJ. Pharmacokinetic studies indicate that inges-
tion of OJ and other citrus fruits and juices may give rise to
Downloaded by [UNISC] at 15:03 06 August 2013
6 S. I. R. FRANKE ET AL.
tissue levels of flavonoids sufficient to exert a biological effect
in humans, on the order of 0.5 μM naringenin in plasma and
10 μM naringenin in liver. In agreement with this, an ingestion
of 400–760 mL OJ was shown to result in 0.6 μM naringenin
in serum (84).
Hesperidin 25 mg/kg body weight was shown to protect
lung carcinogenesis in mice (85) and, at 1000 ppm in the
diet, was shown to reduce rat oral (86, 87), esophageal (88),
urinary-bladder (89), and colon (90) carcinogenesis. There is
evidence of an anticancer effect of hesperidin in skin tumors in
mice, mainly in the promotion stage but not in cancer initiation
(91, 92), and much of its effect has been associated with the inhi-
bition of cell cycle progression (89). Hesperidin has been used as
a pharmaceutical because of its ability to improve the permeabil-
ity and integrity of the capillary lining (93). Hesperidin action
(200 mg/kg oral administration) in acute lung injury involves
the suppression of inflammation, as seen by the downregula-
tion of many inflammatory interleukins, chemokines, adhesion
molecules, and nitric oxide and leukocyte infiltration, as well as
the inhibition of the phosphorylation of IκB, a blocker of NFκB,
p38
MAPK
and JNK
MAPK
(94). Hesperidin (10 and 25 mg/kg) was
shown to ameliorate colonic inflammation by reducing colonic
damage and colonic mieloperoxidase activity in animal models
of inflammatory colitis (95). Hesperidin (5–100 μM in a dose-
dependent fashion) was shown to inhibit mitogenic stimulation
of aortic vascular smooth cells by arresting the cells with no
apoptosis. Cell cycle arrest was associated with an upregulation
of p27
kip1
in parallel with the downregulation of retinoblastoma
protein, cyclins, and proliferating cell nuclear antigen, but there
were no changes in MAPK (96). Hesperidin (10 and 100 μM
in a concentration-dependent fashion) also seems to induce cy-
totoxicity of colon cancer cells through the downregulation of
B-cell CLL/lymphoma 2 (BCL2) and upregulation of caspase-3
and Bcl-2–associated X protein (Bax) (97). Hesperidin in vitro
(50–200 μM) and in vivo (25 mg/kg) was shown to contribute
against dysplasia by inhibition cell hepatocytes invasion and
downregulating metalloproteinase expression, a family of en-
zymes involved in extracellular matrix degradation (98, 99).
Similarly, naringenin in vitro (0.01–0.3 μM) and in vivo (50
and 100 mg/kg, in a dose-dependent manner) has been shown
to induce antiinflammatory effects in different cells (100, 101).
Both naringenin and hesperidin (0.01–0.3 μM) inhibited TNF-α
production at similar levels in glial cells, but naringenin inhib-
ited iNOS in a concentration-dependent fashion, whereas hes-
peridin was not effective in inhibiting the enzyme. Naringenin
but not hesperidin was also found to reduce cell mortality and
inhibit the phosphorylation of p38
MAPK
and STAT-1a (a member
of the signal transducers and activator of transcription family,
which have a role in immune maturation and tolerance as well
as in tumor surveillance). The exclusive feature of naringenin
regarding the presence of the hydroxyl group in the B-ring in-
stead of a catechol, as in all other flavonoids, might explain this
activity (100). Naringin and naringenin (10–50 μM), conversely
to other flavanones, were not capable of reducing viability,
invasion, motility, and cell-matrix adhesion in human lung ade-
nocarcinoma cells (102). Interestingly, a bitter orange extract
rich in naringin and neohesperition was shown to repress plas-
minogen activator inhibitor 1 (PAI-1), a serine protease inhibitor,
and to upregulate matrix metallopeptidase 12, a macrophage
elastase, in human colon fibroblasts treated with TNF-α but not
in controls. This suggests different effects on healthy and dis-
eased cells, which may be beneficial in healthy cells to prevent
sustained inflammation (93).
Beyond flavonoids, OJ has been used as a source of various
micronutrients and phytochemicals with or without the addition
of vitamins and other compounds as supplements (103, 104).
Therefore, the many other constituents of OJ should not be
overlooked.
Limonoids are strong antioxidants (105) that have antipro-
liferative effects (56). Recent studies have indicated that D-
limonene is a potential chemotherapeutic agent, but this drug’s
mechanism remains to be elucidated. The available evidence in-
dicates that D-limonene induces apoptosis via the mitochondrial
death pathway and the suppression of the PI3K/Akt pathway
(106). D-limonene also seems to block angiogenesis by inhibit-
ing vascular endothelial growth factor and to block metastasis
by reducing the expression of matrix metalloproteinases (107).
Coumarins (108) and bioactive amines (7) are also present
in OJ. Beyond studies of the antioxidant properties of cer-
tain coumarins (109) and bioactive amines (110), the results
of further studies regarding the chemopreventive actions of OJ-
specific compounds are still pending.
The vitamin C content of 500 mL OJ is approximately 250 mg
(111), which can lead to approximately 70 μM vitamin C in
the serum (112). Vitamin C protects against the occurrence of
several types of cancer, including mouth, esophageal, gastric,
pancreatic, and rectal cancer (113–115).
Another compound for which OJ is a key biological source
is folate. For instance, 500 mL OJ can provide 150 μg folate
(111), nearly 40% of the Dietary Reference Intake for the nu-
trient (116). OJ is among the major food sources of folate on
a given day for the U.S. population (117). Folate is involved in
DNA synthesis, repair. and methylation (11). Folate deficiency
causes uracil misincorporation into human DNA and chromo-
some breakage (118). The folate status modulates the risk of de-
veloping cancers in selected tissues, the most notable of which
is the colorectum (119).
POTENTIAL TOXICITY OF OJ
General Effects
OJ intake can generate noxious effects for some individuals,
particularly in large amounts. For example, excessive amounts
of OJ might induce hyperkalemia, especially in renally com-
promised individuals (120). OJ has been controversially linked
to the risk of food allergy in sensitive individuals (121, 122).
OJ was found to stimulate salivary secretion, decrease the sali-
vary pH immediately after consumption, and decrease the redox
Downloaded by [UNISC] at 15:03 06 August 2013
ORANGE JUICE AND CANCER 7
potential of whole saliva. This can be caries-promoting in the
oral cavity in persons with low salivary flow rate who consume
the juice regularly (123). Moreover, sour (Seville) orange (Cit-
rus auratium) extracts have been shown to induce weight gain
and increase blood pressure and cardiac dysfunctions in hyper-
tensive subjects, possibly because of its synephrine, octopamine,
and furocumarin content. Indeed, sour orange juice (SOJ) should
be avoided by individuals with severe hypertension or glau-
coma and by those taking monoamine oxidase inhibitors. SOJ
consumption seems to be safe for normotensive subjects (124).
OJ should be considered as a part of the recommended daily
ingestion of at least five portions of fruit and vegetables to pre-
vent cancer in healthy adults (125). However, the recommended
daily portions for intake of fruit juices among children and hy-
pertensive, kidney-compromised, or diabetic subjects could be
lower than recommended for healthy people. The use of OJ as
a substitute for meals can lead to malnutrition and decreased
stature in children (126). Excessive intake of any food, even for
the healthiest, can lead to oxidative status imbalance.
Unpasteurized OJ is a known vehicle for salmonellosis, and
several cases of outbreaks of bacterial infections were reported
in the United States (127).
Prooxidant Effect, Genotoxicity, and Carcinogenicity
of OJ: Is There Any Harm?
Despite scientific data reporting beneficial properties derived
from the consumption of OJ (for example, antimutagenic or
anticarcinogenic effects), some compounds also present in OJ
have been identified as being mutagenic or carcinogenic (5, 42,
128). The carcinogenic or genotoxic effects of OJ might be
mediated by the interaction of juice components with transition
metals or by sub-products of juice autooxidation. Some in vitro
and in vivo studies report prooxidant and genotoxic effects for
OJ. For example, Franke et al. (4) showed significant increases
in lipid peroxidation induced by OJ in vitro. In vivo results also
showed an increase of lipid peroxidation in the serum of healthy
volunteers 2 h after the intake of OJ but only for 8-day-aged
juice stored at 4
C (32).
There is evidence of OJ mutagenicity in bacteria. Mazaki
et al. (129) observed mutagenic activity in Salmonella/
microsome assay. Heated OJ, as well as the acid hydrolysate,
were mutagenic and cytotoxic in Salmonella typhimurium with-
out S-9-mix (hepatic microsome fraction, added to simulate
mammalian metabolism), after neutralization to pH 7.4 (130,
131). More recent data of Friedman et al. (132) did not confirm
the dramatic increase in mutagenicity reported for heated OJ,
and the response without S-9 activation was similar for juices
either heated or not, ranging from 2 to 3 times the background
values. In agreement with the previous results, Franke et al. (4)
evaluated the mutagenic activity of frozen and fresh forms of
in natura and processed OJ (pH not adjusted) using 4 strains
of Salmonella in the Ames test with or without S-9-mix. Only
1 unsweetened, unfrozen, and processed OJ sample was not
mutagenic for any of the strains tested. This OJ also had the
highest antioxidant activity. Interestingly, the content of vita-
min C and phenolic compounds correlated to the mutagenicity
(4). Although heating, normally used in processing OJ, is in-
sufficient to release mutagenic flavonol as aglycones from their
glycosides (kaempherol and quercetin), other compounds such
as Maillard intermediary product (130) or mutagenic brown-
ing products derived from free lysine, histidine, or other amino
acids (132) might be generated. As an alternative hypothesis
to OJ mutagenicity, one should recall that plant-derived com-
pounds have antibacterial activity (133), which might partially
explain the OJ mutagenicity in bacteria.
In mammals there is no evidence of the genotoxicity of OJ.
Nevertheless, higher doses of OJ (equivalent to 600 mL in
humans) can induce a slight transient increase in DNA damage
in blood cells of mice, as detected by Franke et al. (41, 42).
Evidence linking OJ consumption to cancer risk is sparse and
controversial. It has been recently hypothesized that cutaneous
melanoma grows as the availability and consumption of citrus
products increases, which may be related to concomitant in-
creases in dietary photocarcinogenic furocoumarins (134). In
support of this hypothesis, Feskanich et al. (15), studying 2
Nurse’s Health Study Cohorts, found an unexpected associa-
tion between the increased risk of melanoma and higher intake
of food vitamin C, particularly from OJ. The authors describe
this association as random but also state that the association
was strongest among the higher-risk, sun-sensitive women. It
is unlikely that vitamin C was responsible for the association,
because supplements of vitamin C were not associated with a
higher risk of melanoma. OJ contains appreciable quantities of
furocoumarins such as psoralens. Psoralens are substances that
cause interstrand cross-links and monoadditions in DNA when
photoactivated by UVA (135). On the contrary, the case-control
study of Hakim et al (14) with elder from Arizona indicated
that the intake of citrus peel reduced the risk of squamous cell
carcinoma of the skin, pointing out an intake-dependent chemo-
preventive effect of substances present in citrus peel such as
d-limonene. Considering this data, epidemiological studies eval-
uating either the beneficial or noxious effect of fruit juices are
needed, particularly for those taking into account differences
in the processing and composition of these drinks, as well as
controlling for confounding variables, such as lifestyle and diet,
that may lead to false interpretations.
WEIGHT OF EVIDENCE REGARDING CANCER
CHEMOPREVENTION BY OJ
To date, no epidemiological study specifically evaluated the
chemopreventive action of OJ in humans. There are few studies
using rodent models of cancer that evaluated the effect of OJ
on cancer chemoprevention. The results from the rodent cancer
models revealed relevant cancer chemoprevention evidence for
OJ according to the “hierarchy of robustness,” as defined by the
Joint Panel of the World Cancer Research Fund and the Amer-
ican Institute for Cancer Research (136) for evaluating cancer
Downloaded by [UNISC] at 15:03 06 August 2013
8 S. I. R. FRANKE ET AL.
chemoprevention. The Panel ranks human and animal experi-
mental studies in evaluating the role of dietary and physical ac-
tivity in the risk/prevention of human cancer. Class 1 evidence
refers to 1) in vivo data from controlled human feeding studies,
2) data from genetically modified models of human diseases,
and 3) in vivo studies using rodent cancer models designed to
investigate modifiers of the cancer process (136). The available
evidence can be classified as Class 1c because of the studies
using rodent models of cancer, summarized in Table 1.
Several biological effects that can contribute to chemopre-
vention were shown for OJ. In this review, we summarized
several of these effects, including antioxidant, antimutagenic
and antigenotoxic, cytoprotective, hormonal,and cell signaling-
modulating effects. OJ has antimicrobial and antiviral action and
modulates the absorption of xenobiotics. Therefore, OJ could
contribute to chemoprevention at every stage of cancer initia-
tion and progression. Among the most relevant biological effects
of OJ is the juice’s antigenotoxic and antimutagenic potential,
which was shown in cells in culture and in rodents and humans.
The biological effects of OJ in vitro were shown to be largely
influenced by the juice’s composition. The composition of OJ
depends on physiological conditions (related to climate, soil
and fruit maturation, the genetic characteristics (varietal) of the
oranges and variations in processing methods and storage times
and methods. The addition of sugars seems to substantially de-
crease the antioxidant effect of OJ. Thermal treatments, storage
above 20
C or both can lead to an even greater decrease in
antioxidant activity.
There are numerous studies evaluating the biological effects
of the constituents of OJ. Although many of these components
are chemopreventive agents in isolation, it is difficult to under-
stand how the compounds interact when ingested as part of OJ.
It is very likely that the individual constituents of OJ act syner-
gistically and antagonistically rather than simply by an additive
mechanism.
Epidemiological studies and randomized controlled interven-
tion studies in humans evaluating the chemopreventive effect
of OJ, in terms of amount, consistency, and quality, on can-
cer risk are needed to establish this juice’s chemopreventive
effect. The type and amount of OJ to be tested should be de-
fined with caution, as excessive intake of OJ can be noxious to
children (cavities and undernutrition may occur if OJ is used
as a main meal substitute) or hypertensive (increases in sour
OJ consumption could lead to a pressure increase and cardiac
dysfunction), renally compromised (hyperkalemic), or diabetic
(hyperglycemic) subjects. OJ should be also pasteurized given
the risk of bacterial outbreaks.
ACKNOWLEDGMENTS
The authors thank Drs. Christine Gaylarde and Jaqueline
Nascimento Picada for critically reading the manuscript. The
research was funded by FAPERGS and the University of Santa
Cruz do Sul.
REFERENCES
1. Morton J: Orange.In:Fruits of Warm Climates, J.F. Morton (ed.). Miami,
FL: Florida Flair Books, 1987, 134–142.
2. Moufida S and Marzouk B: Biochemical characterization of blood orange,
sweet orange, lemon, bergamot and bitter orange. Phytochemistry 62,
1283–1289, 2003.
3. Fiore A, Fauci LL, Cervellati R, Guerra MC, Speroni E, et al.: Antioxidant
activity of pasteurized and sterilized commercial red orange juices. Mol
Nutr Food Res 49, 1129–1135, 2005.
4. Franke SIR, Ckless K, Silveira JD, Rubensam G, Brendel M, et al.: Study
of antioxidant and mutagenic activity of different orange juices. Food
Chem 88, 45–55, 2004.
5. Patrineli A, Clifford MN, Walker R, and Ioannides C: Mutagenicity of
white grape juice in the Ames test. Food Chem Toxicol 34, 559–562,
1996.
6. Franke AA, Cooney RV, Henning SM, and Custer LJ: Bioavailability and
Antioxidant effects of orange juice components in humans. J Agric Food
Chem 53, 5170–5178, 2005.
7. Vieira SM, Theodoro KH, and Gloria MBA: Profile and levels of bioac-
tive amines in orange juice and orange soft drink. Food Chemistry 100,
895–903, 2007.
8. Rampersaud GC: A comparison of nutrient density scores for 100% fruit
juices. Journal of Food Science 72, S261–S266, 2007.
9. O’ Neil, CE, Nicklas TA, Rampersaud GC, and Fulgoni VL, III: 100%
Orange juice consumption is associated with better diet quality, improved
nutrient adequacy, decreased risk for obesity, and improved biomarkers
of health in adults: National Health and Nutrition Examination Survey,
2003–2006. Nutr J 11, 107, 2012.
10. Halliwell B: Vitamin C and genomic stability. Mutat Res 475, 29–35,
2001.
11. Fenech M and Crott JW: Micronuclei, nucleoplasmic bridges and nu-
clear buds induced in folic acid deficient human lymphocytes-evidence
for breakage–fusion–bridge cycles in the cytokinesis-block micronucleus
assay. Mutat Res 504, 131–136, 2002.
12. Middleton E, Jr., Kandaswami C, and Theoharides TC: The effects of
plant flavonoids on mammalian cells: implications for inflammation, heart
disease, and cancer. Pharmacol Rev 52, 673–751, 2000.
13. Ercisli S and Ohran E: Chemical composition of withe (Morus alba), red
(Morus rubra) and black (Morus nigra) mulberry fruits. Food Chemistry
103, 1380–1384, 2007.
14. Hakim IA, Harris RB, and Ritenbaugh C: Citrus peel use is associated
with reduced risk of squamous cell carcinoma of the skin. Nutr Cancer
37, 161–168, 2000.
15. Feskanich D, Willett WC, Hunter DJ, and Colditz GA: Dietary intakes of
vitamins A, C, and E and risk of melanoma in two cohorts of women. Br
J Cancer 88, 1381–1387, 2003.
16. Kwan ML, Block G, Selvin S, Month S, and Buffler PA: Food consumption
by children and the risk of childhood acute leukemia. Am J Epidemiol 160,
1098–1107, 2004.
17. So FV, Guthrie N, Chambers AF, Moussa M, and Carroll KK: Inhibition
of human breast cancer cell proliferation and delay of mammary tumori-
genesis by flavonoids and citrus juices. Nutr Cancer 26, 167–181, 1996.
18. He YP, Root MM, Parker RS, and Campbell TC: Effects of carotenoid-
rich food extracts on the development of preneoplastic lesions in rat liver
and on in vivo and in vitro antioxidant status. Nutrition and Cancer 27,
238–244, 1997.
19. Miyagi Y, Om AS, Chee KM, and Bennink MR: Inhibition of
azoxymethane-induced colon cancer by orange juice. Nutr Cancer 36,
224–229, 2000.
20. Guthrie N and Carroll KK:”Inhibition of mammary cancer by citrus
flavonoids. Adv Exp Med Biol 439, 227–236, 1998.
21. Lichtenthaler R and Marx F: Total oxidant scavenging capacities of com-
mon European fruit and vegetable juices. Journal of Agricultural and Food
Chemistry 53, 103–110, 2005.
Downloaded by [UNISC] at 15:03 06 August 2013
ORANGE JUICE AND CANCER 9
22. Proteggente AR, Pannala AS, Paganga G, Van Buren L, Wagner E, et al.:
The antioxidant activity of regularly consumed fruit and vegetables reflects
their phenolic and vitamin C composition. Free Radic Res 36, 217–233,
2002.
23. Del Caro A, Piga A, Vacca V, and Agabbio M: Changes of flavonoids,
vitamin C and antioxidant capacity in minimally processed citrus segments
and juices during storage. Food Chemistry 84, 99–105, 2004.
24. Lo Scalzo R, Iannoccari T, Summa C, Morelli R, and Rapisarda P: Effect of
thermal treatments on antioxidant and antiradical activity of blood orange
juice. Food Chemistry 85, 41–47, 2004.
25. Klimczak, I, Malcka, M, Szlachta, M, and Gliszczynska-Swiglo, A: Effect
of storage on the content of polyphenols, vitamin C and the antioxidant
activity of orange juices. J Food Compost Anal 20, 313–322, 2007.
26. Plaza L, S
´
anchez-Moreno C, Elez-Mart
´
ınez P, Ancos BD, Mart
´
ın-Belloso
O, et al.: Effect of refrigerated storage on vitamin C and antioxidant activity
of orange juice processed by high-pressure or pulsed electric fields with
regard to low pasteurization. Eur Food Res Technol 223, 487–493, 2006.
27. Rapisarda P, Bianco ML, Pannuzzo P, and Timpanaro N: Effect of cold
storage on vitamin C, phenolics and antioxidant activity of five orange
genotypes [Citrus sinensis (L.) Osbeck]. Postharvest Biology and Tech-
nology 49, 348–354, 2008.
28. Choi MH, Kim GH, and Lee HS: Effects of ascorbic acid retention on juice
color and pigment stability in blood orange (Citrus sinensis) juice during
refrigerated storage. Food Research International 35, 753–759, 2002.
29. Sentandreu E, Navarro JL, and Sendra JM: Effect of technological pro-
cesses and storage on flavonoids content and total, cumulative fast-kinetics
and cumulative slow-kinetics antiradical activities of citrus juices. Eur
Food Res Technol 225, 905–912, 2007.
30. Tabart J, Kevers C, Pincemail J, Defraigne JO, and Dommes J: Compar-
ative antioxidant capacities of phenolic compounds measured by various
tests. Food Chemistry 113, 1226–1233, 2009.
31. Ko SH, Choi SW, Ye SK, Cho BL, Kim HS, et al.: Comparison of the
antioxidant activities of nine different fruits in human plasma. Journal of
Medicinal Food 8, 41–46, 2005.
32. Johnston CS and Hale JC: Oxidation of ascorbic acid in stored orange juice
is associated with reduced plasma vitamin C concentrations and elevated
lipid peroxides. J Am Diet Assoc 105, 106–109, 2005.
33. Ghanim H, Mohanty P, Pathak R, Chaudhuri A, Sia CL, et al.: Orange juice
or fructose intake does not induce oxidative and inflammatory response.
Diabetes Care 30, 1406–1411, 2007.
34. Sanchez-Moreno C, Cano MP, de Ancos B, Plaza L, Olmedilla B, et al.:
High-pressurized orange juice consumption affects plasma vitamin C,
antioxidative status and inflammatory markers in healthy humans. JNutr
133, 2204–2209, 2003.
35. Riso P, Visioli F, Gardana C, Grande S, Brusamolino A, et al.: Effects of
blood orange juice intake on antioxidant bioavailability and on different
markers related to oxidative stress. J Agric Food Chem 53, 941–947, 2005.
36. Gorinstein S, Leontowicz H, Leontowicz M, Krzeminski R, Gralak M,
et al.: Fresh Israeli Jaffa blond (Shamouti) orange and Israeli Jaffa red
Star Ruby (Sunrise) grapefruit juices affect plasma lipid metabolism and
antioxidant capacity in rats fed added cholesterol. J Agric Food Chem 52,
4853–4859, 2004.
37. Maeda N, Hagihara H, Nakata Y, Hiller S, Wilder J, et al.: Aortic wall
damage in mice unable to synthesize ascorbic acid. Proc Natl Acad Sci
USA97, 841–846, 2000.
38. Bala S and Grover IS: Antimutagenicity of some citrus fruits in Salmonella
typhimurium. Mutat Res 222, 141–148, 1989.
39. He Y and Campbell TC: Effects of carotenoids on aflatoxin B1-induced
mutagenesis in S. typhimurium TA 100 and TA 98.
Nutr Cancer 13,
243–253, 1990.
40. Higashimoto M, Yamato H, Kinouchi T, and Ohnishi Y: Inhibitory effects
of citrus fruits on the mutagenicity of 1-methyl-1,2,3,4-tetrahydro-beta-
carboline-3-carboxylic acid treated with nitrite in the presence of ethanol.
Mutat Res 415, 219–226, 1998.
41. Franke SIR, Pra D, Erdtmann B, Henriques JAP, and da Silva J: Influence
of orange juice over the genotoxicity induced by alkylating agents: an in
vivo analysis. Mutagenesis 20, 279–283, 2005.
42. Franke SIR, Pr
´
a D, Dias JF, Giulian R, Yoneama ML, et al.: Influence of
orange juice in the levels and in the genotoxicity of iron and copper. Food
Chem Toxicol 44, 425–435, 2006.
43. Guarnieri S, Riso P, and Porrini M: Orange juice vs vitamin C: effect
on hydrogen peroxide-induced DNA damage in mononuclear blood cells.
British Journal of Nutrition 97, 639–643, 2007.
44. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, et al.: Single
cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology
testing. Environ Mol Mutagen 35, 206–221, 2000.
45. Hosseinimehr SJ and Karami M: Citrus extract modulates genotoxicity
induced by cyclophosphamide in mice bone marrow cells. J Pharm Phar-
macol 57, 505–509, 2005.
46. Hosseinimehr SJ, Tavakoli H, Pourheidari G, Sobhani A, and Shafiee
A: Radioprotective effects of citrus extract against gamma-irradiation in
mouse bone marrow cells. JRadRes44, 237–241, 2003.
47. Ohsawa K, Nakagawa SY, Kimura M, Shimada C, Tsuda S, et al.: Detec-
tion of in vivo genotoxicity of endogenously formed N-nitroso compounds
and suppression by ascorbic acid, teas and fruit juices. Mutat Res 539,
65–76, 2003.
48. Coussens LM and Werb Z: Inflammation and cancer. Nature 420, 860–867,
2002.
49. O’Reilly JD, Mallet AI, McAnlis GT, Young IS, Halliwell B, et al.: Con-
sumption of flavonoids in onions and black tea: lack of effect on F2-
isoprostanes and autoantibodies to oxidized LDL in healthy humans. Am
J Clin Nutr 73, 1040–1044, 2001.
50. Ekmekcioglu C, Strauss-Blasch, G, Leibetseder V, and Marktl W: Toxico-
logical and biochemical effects of different beverages on human intestinal
cells. Food Research International 32, 421, 1999.
51. Lim SL and Lim LY: Effects of citrus fruit juices on cytotoxicity and drug
transport pathways of Caco-2 cell monolayers. International Journal of
Pharmaceutics 307, 42–50, 2006.
52. Vitali F, Pennisi C, Tomaino A, Bonina F, De Pasquale A, et al.: Effect
of a standardized extract of red orange juice on proliferation of human
prostate cells in vitro. Fitoterapia 77, 151–155, 2006.
53. Park HJ, Kim MJ, Ha E, and Chung JH: Apoptotic effect of hesperidin
through caspase3 activation in human colon cancer cells, SNU-C4”. Phy-
tomedicine 15, 147–151, 2008.
54. Kang JS, Cho D, Kim YI, Hahm E, Kim YS, et al.: Sodium ascorbate
(vitamin C) induces apoptosis in melanoma cells via the down-regulation
of transferrin receptor dependent iron uptake. J Cell Physiol 204, 192–197,
2005.
55. Duarte TL, Cooke MS, and Jones GD: Gene expression profiling reveals
new protective roles for vitamin C in human skin cells. Free Radic Biol
Med 46, 78–87, 2009.
56. Tian Q, Miller EG, Ahmad H, Tang L, and Patil BS: Differential inhibition
of human cancer cell proliferation by citrus limonoids. Nutr Cancer 40,
180–184, 2001.
57. Wietrzyk J, Grynkiewicz G, and Opolski A: Phytoestrogens in cancer pre-
vention and therapy—mechanisms of their biological activity
. Anticancer
Res 25, 2357–2366, 2005.
58. Horn-Ross PL, Lee M, John EM, and Koo J: Sources of phytoestrogen
exposure among non-Asian women in California, USA. Cancer Causes
Control 11, 299–302, 2000.
59. Philp HA: Hot flashes—a review of the literature on alternative and com-
plementary treatment approaches. Altern Med Rev 8, 284–302, 2003.
60. Deyhim F, Garica K, Lopez E, Gonzalez J, Ino S, et al.: Citrus juice
modulates bone strength in male senescent rat model of osteoporosis.
Nutrition 22, 559–563, 2006.
61. Selgrad M, Malfertheiner P, Fini L, Goel A, Boland CR, et al.: The role
of viral and bacterial pathogens in gastrointestinal cancer. J Cell Physiol
216, 378–388, 2008.
Downloaded by [UNISC] at 15:03 06 August 2013
10 S. I. R. FRANKE ET AL.
62. Baird IM, Hughes RE, Wilson HK, Davies JE, and Howard AN: The effects
of ascorbic acid and flavonoids on the occurrence of symptoms normally
associated with the common cold. Am J Clin Nutr 32, 1686–1690, 1979.
63. Hemil
¨
a H, Chalker E, and Douglas B: Vitamin C for preventing and
treating the common cold. Cochrane Database of Systematic Reviews 3,
2007. doi:10.1002/14651858.CD000980.pub3.
64. Zafriri D, Ofek I, Adar R, Pocino M, and Sharon N: Inhibitory activity of
cranberry juice on adherence of type 1 and type P fimbriated Escherichia
coli to eucaryotic cells. Antimicrob Agents Chemother 33, 92–98, 1989.
65. Hazell T and Johnson IT: In vitro estimation of iron availability from a
range of plant foods: influence of phytate, ascorbate and citrate. Br J Nutr
57, 223–233, 1987.
66. Fairweather-Tait S, Hickson K, McGaw B, and Reid M: Orange juice
enhances aluminium absorption from antacid preparation. Eur J Clin Nutr
48, 71–73, 1994.
67. Mehansho H, Kanerva RL, Hudepohl GR, and Smith KT: Calcium
bioavailability and iron–calcium interaction in orange juice. JAmColl
Nutr 8, 61–68, 1989.
68. Flanagan PR, Cluett J, Chamberlain MJ, and Valberg LS: Dual-isotope
method for determination of human zinc absorption: the use of a test meal
of turkey meat. JNutr115, 111–122, 1985.
69. Robinson MF, Thomson CD, and Huemmer PK: Effect of a megadose of
ascorbic acid, a meal and orange juice on the absorption of selenium as
sodium selenite. NZMedJ98, 627–629, 1985.
70. El-Shobaki FA, El-Hawary Z, Saleh N, and Morcos SR: Effect of au-
thentic and natural vitamin C and carotene on amino acid absorption. Z
Ernahrungswiss 16, 12–17, 1977.
71. Odou P, Barthelemy C, and Robert H: Influence of seven beverages on
salicylate disposition in humans. J Clin Pharm Ther 26, 187–193, 2001.
72. Nishimuta H, Tsujimoto M, Ogura K, Hiratsuka A, Ohtani H, et al.: In-
hibitory effects of various beverages on ritodrine sulfation by recombinant
human sulfotransferase isoforms SULT1A1 and SULT1A3. Pharm Res
22, 1406–1410, 2005.
73. Lilja JJ, Raaska K, and Neuvonen PJ: Effects of orange juice on the
pharmacokinetics of atenolol. Eur J Clin Pharmacol 61, 337–340, 2005.
74. Kamath AV, Yao M, Zhang Y, and Chong S: Effect of fruit juices on
the oral bioavailability of fexofenadine in rats. Journal of Pharmaceutical
Sciences 94, 233–239, 2005.
75. Penzak SR, Acosta EP, Turner M, Edwards DJ, Hon YY, et al.: Effect of
Seville orange juice and grapefruit juice on indinavir pharmacokinetics. J
Clin Pharmacol 42, 1165–1170, 2002.
76. Ikegawa T, Ushigome F, Koyabu N, Morimoto S, Shoyama Y, et al.: Inhibi-
tion of P-glycoprotein by orange juice components, polymethoxyflavones
in adriamycin-resistant human myelogenous leukemia (K562/ADM) cells.
Cancer Lett 160, 21–28, 2000.
77. Satoh H, Yamashita F, Tsujimoto M, Murakami H, Koyabu N, et al.:
Citrus juices inhibit the function of human organic anion-transporting
polypeptide OATP-B. Drug Metab Dispos 33, 518–523, 2005.
78. Saito M, Hirata-Koizumi M, Matsumoto M, Urano T, and Hasegawa R:
Undesirable effects of citrus juice on the pharmacokinetics of drugs: focus
on recent studies. Drug Saf 28, 677–694, 2005.
79. Brand W, Schutte ME, Williamson G, van Zanden JJ, Cnubben NHP,
et al.: Flavonoid-mediated inhibition of intestinal ABC transporters may
affect the oral bioavailability of drugs, food-borne toxic compounds and
bioactive ingredients. Biomedicine & Pharmacotherapy
60, 508–519,
2006.
80. Miller EG, Peacock JJ, Bourland TC, Taylor SE, Wright JA, et al.: Inhibi-
tion of oral carcinogenesis by citrus flavonoids. Nutrition and Cancer 60,
69–74, 2008.
81. Shen HM and Tergaonkar V: NFkappaB signaling in carcinogenesis and
as a potential molecular target for cancer therapy. Apoptosis 14, 348–363,
2009.
82. Bradham C and McClay DR: p38 MAPK in development and cancer. Cell
Cycle 5, 824–828, 2006.
83. Wu WS, Wu JR, and Hu CT: Signal cross talks for sustained MAPK
activation and cell migration: the potential role of reactive oxygen species.
Cancer Metastasis Rev 27, 303–314, 2008.
84. Breinholt VM, Svendsen GW, Dragsted LO, and Hossaini A: Citrus-
derived flavonoid naringenin exerts uterotrophic effects in female mice at
human relevant doses. Basic & Clinical Pharmacology & Toxicology 94,
30–36, 2004.
85. Kamaraj S, Ramakrishnan G, Anandakumar P, Jagan S, and Devaki T: An-
tioxidant and anticancer efficacy of hesperidin in benzo(a)pyrene induced
lung carcinogenesis in mice. Investigational New Drugs 27, 214–222,
2009.
86. Tanaka T, Makita H, Ohnishi M, Hirose Y, Wang A, et al.: Chemopreven-
tion of 4-nitroquinoline 1-oxide-induced oral carcinogenesis by dietary
curcumin and hesperidin: comparison with the protective effect of beta-
carotene. Cancer Res 54, 4653–4659, 1994.
87. Tanaka T, Makita H, Ohnishi M, Mori H, Satoh K, et al.: Chemopre-
vention of 4-nitroquinoline 1-oxide-induced oral carcinogenesis in rats by
flavonoids diosmin and hesperidin, each alone and in combination. Cancer
Res 57, 246–252, 1997.
88. Tanaka T, Makita H, Kawabata K, Mori H, Kakumoto M, et al.: Modu-
lation of N-methyl-N-amylnitrosamine-induced rat oesophageal tumouri-
genesis by dietary feeding of diosmin and hesperidin, both alone and in
combination. Carcinogenesis 18, 761–769, 1997.
89. Yang M, Tanaka T, Hirose Y, Deguchi T, Mori H, et al.:
Chemopreventive effects of diosmin and hesperidin on N-butyl-N-
(4-hydroxybutyl)nitrosamine-induced urinary-bladder carcinogenesis in
male ICR mice. Int J Cancer 73, 719–724, 1997.
90. Tanaka T, Makita H, Kawabata K, Mori H, Kakumoto M, et al.: Chemo-
prevention of azoxymethane-induced rat colon carcinogenesis by the nat-
urally occurring flavonoids, diosmin and hesperidin. Carcinogenesis 18,
957–965, 1997.
91. Berkarda B, Koyuncu H, Soybir G, and Baykut F: Inhibitory effect of
Hesperidin on tumour initiation and promotion in mouse skin. Research
in Experimental Medicine 198, 93–99, 1998.
92. Koyuncu H, Berkarda B, Baykut F, Soybir G, Alatli C, et al.: Pre-
ventive effect of hesperidin against inflammation in CD-1 mouse
skin caused by tumor promoter. Anticancer Research 19, 3237–3241,
1999.
93. Gimenez-Bastida JA, Martinez-Florensa M, Espin JC, Tomas-Barberan
FA, and Garcia-Conesa MT: A citrus extract containing flavanones re-
presses plasminogen activator inhibitor-1 (PAI-1) expression and regulates
multiple inflammatory, tissue repair, and fibrosis genes in human colon
fibroblasts. Journal of Agricultural and Food Chemistry 57, 9305–9315,
2009.
94. Yeh CC, Kao SJ, Lin CC, Wang SD, Liu CJ, et al.: The immunomodulation
of endotoxin-induced acute lung injury by hesperidin in vivo and in vitro.
Life Sciences 80, 1821–1831, 2007.
95. Crespo ME, Galvez J, Cruz T, Ocete MA, and Zarzuelo A: Anti-
inflammatory activity of diosmin and hesperidin in rat colitis induced
by TNBS. Planta Medica 65, 651–653, 1999.
96. Jin YR, Han XH, Zhang YH, Lee JJ, Lim Y, et al.: Hesperetin, a
bioflavonoid, inhibits rat aortic vascular smooth muscle cells prolifera-
tion by arresting cell cycle. Journal of Cellular Biochemistry 104, 1–14,
2008.
97. Park HJ, Kim MJ, Ha E, and Chung JH: Apoptotic effect of hesperidin
through caspase3 activation in human colon cancer cells, SNU-C4. Phy-
tomedicine
15, 147–151, 2008.
98. Yeh M-H, Kao S-T, Hung C-M, Liu C-J, Lee K-H, et al.: Hesperidin inhib-
ited acetaldehyde-induced matrix metalloproteinase-9 gene expression in
human hepatocellular carcinoma cells. Toxicology Letters 184, 204–210,
2009.
99. Balakrishnan A and Menon VP: Effect of hesperidin on matrix met-
alloproteinases and antioxidant status during nicotine-induced toxicity.
Toxicology 238, 90–98, 2007.
Downloaded by [UNISC] at 15:03 06 August 2013
ORANGE JUICE AND CANCER 11
100. Vafeiadou K, Vauzour D, Lee HY, Rodriguez-Mateos A, Williams RJ,
et al.: The citrus flavanone naringenin inhibits inflammatory signalling
in glial cells and protects against neuroinflammatory injury. Archives of
Biochemistry and Biophysics 484, 100–109, 2009.
101. Shi Y, Dai J, Liu H, Li RR, Sun PL, et al.: Naringenin inhibits allergen-
induced airway inflammation and airway responsiveness and inhibits NF-
kappa B activity in a murine model of asthma. Canadian Journal of
Physiology and Pharmacology 87, 729–735, 2009.
102. Hsiao YC, Kuo WH, Chen PN, Chang HR, Lin TH, et al.: Flavanone
and 2’-OH flavanone inhibit metastasis of lung cancer cells via down-
regulation of proteinases activities and MAPK pathway. Chem Biol Inter-
act 167, 193–206, 2007.
103. Economos CD, Biancuzzo R, Hyatt RR, Kuder J, Provias M, et al.: Vitamin
D deficiency is common in young children in Boston: Vitamin D3 in orange
juice was an effective treatment. Bone 40, S52–S52, 2007.
104. Devaraj S, Jialal I, and Vega-Lopez S: Plant sterol-fortified orange juice ef-
fectively lowers cholesterol levels in mildly hypercholesterolemic healthy
individuals. Arterioscler Thromb Vasc Biol 24, e25–e28, 2004.
105. Sun CD, Chen KS, Chen Y, and Chen QJ: Contents and antioxidant
capacity of limonin and nomilin in different tissues of citrus fruit of four
cultivars during fruit growth and maturation. Food Chemistry 93, 599–605,
2005.
106. Jia SS, Xi GP, Zhang M, Chen YB, Lei B, et al.: Induction of apoptosis by
D-limonene is mediated by inactivation of Akt in LS174T human colon
cancer cells. Oncology Reports 29, 349–354, 2013.
107. Murthy KNC, Jayaprakasha GK, and Patil BS: D-limonene rich volatile
oil from blood oranges inhibits angiogenesis, metastasis and cell death in
human colon cancer cells. Life Sciences 91, 429–439, 2012.
108. Dugo P, Russo M, Saro M, Carnovale C, Bonaccorsi I, et al.: Multidimen-
sional liquid chromatography for the determination of chiral coumarins
and furocoumarins in Citrus essential oils. Journal of Separation Science
35, 1828–1836, 2012.
109. Yu J, Wang LM, Walzem RL, Miller EG, Pike LM, et al.: Antioxidant
activity of citrus limonoids, flavonoids, and coumarins. Journal of Agri-
cultural and Food Chemistry 53, 2009–2014, 2005.
110. Das KC and Misra HP: Hydroxyl radical scavenging and singlet oxygen
quenching properties of polyamines. Molecular and Cellular Biochemistry
262, 127–133, 2004.
111. U.S. Department of Agriculture: USDA National Nutrient Database for
Standard Reference, Release 18. U.S. Department of Agriculture, Agricul-
tural Research Service, Nutrient Data Laboratory, 2005. Retrieved from
http://www.nal.usda.gov/fnic/foodcomp
112. Levine M, Conry-Cantilena C, Wang Y, Welch RW, Washko PW, et al.:
Vitamin C pharmacokinetics in healthy volunteers: evidence for a rec-
ommended dietary allowance. Proc Natl Acad Sci USA 93, 3704–3709,
1996.
113. Mayne ST, Risch HA, Dubrow R, Chow WH, Gammon MD, et al.: Nutri-
ent intake and risk of subtypes of esophageal and gastric cancer. Cancer
Epidemiol Biomarkers Prev 10, 1055–1062, 2001.
114. Padayatty SJ, Katz A, Wang Y, Eck P, Kwon O, et al.: Vitamin C as an
antioxidant: evaluation of its role in disease prevention. J Am Coll Nutr
22, 18–35, 2003.
115. Block G: Vitamin C and cancer prevention: the epidemiologic evidence.
Am J Clin Nutr 53, 270S–282S, 1991.
116. Institute of Medicine: Dietary Reference Intakes for Dietary Reference
Intakes for Thiamin, Riboflavin, Niacin, Vitamin B6, Folate, Vitamin B12,
Pantothenic Acid, Biotin, and Choline. Food and Nutrition Board.Wash-
ington, DC: National Academy Press, 1998.
117. Subar AF, Block G, and James LD: Folate intake and food sources in the
US population. Am J Clin Nutr 50, 508–516, 1989.
118. Blount BC, Mack MM, Wehr CM, MacGregor JT, Hiatt RA, et al.: Folate
deficiency causes uracil misincorporation into human DNA and chromo-
some breakage: implications for cancer and neuronal damage. Proc Natl
Acad Sci USA 94, 3290–3295, 1997.
119. Choi SW and Mason JB: Folate and carcinogenesis: an integrated scheme.
JNutr130, 129–132, 2000.
120. Fan K and Leehey DJ: Orange juice-induced hyperkalemia in a dia-
betic patient with chronic renal failure. Diabetes Care 19, 1457–1458,
1996.
121. Zhu SL, Ye ST, and Yu Y: Allergenicity of orange juice and orange seeds:
a clinical study. Asian Pac J Allergy Immunol 7, 5–8, 1989.
122. Yap JC, Wang YT, Yeo CT, and Poh SC: The effect of fresh orange juice
on bronchial hyperreactivity in asthmatic subjects. Singapore Med J 31,
583–586, 1990.
123. Tenovuo J and Rekola M: Some effects of sugar-flavored acid beverages on
the biochemistry of human whole saliva and dental plaque. Acta Odontol
Scand 35, 317–330, 1977.
124. Penzak SR, Jann MW, Cold JA, Hon YY, Desai HD, et al.: Seville (sour)
orange juice: synephrine content and cardiovascular effects in normoten-
sive adults. J Clin Pharmacol 41, 1059–1063, 2001.
125. McCall MR and Frei B: Can antioxidant vitamins materially reduce
oxidative damage in humans? Free Radic Biol Med 26, 1034–1053,
1999.
126. Dennison BA, Rockwell HL, Nichols MJ, and Jenkins P: Children’s
growth parameters vary by type of fruit juice consumed. J Am Coll Nutr
18, 346–352, 1999.
127. Jain S, Bidol SA, Austin JL, Berl E, Elson F, et al.: Multistate outbreak of
salmonella typhimurium and saintpaul infections associated with unpas-
teurized orange juice—United States, 2005. Clinical Infectious Diseases
48, 1065–1071, 2009.
128. Fenech M, Baghurst P, Luderer W, Turner J, Record S, et al.: Low intake
of calcium, folate, nicotinic acid, vitamin E, retinol, beta-carotene and
high intake of pantothenic acid, biotin and riboflavin are significantly
associated with increased genome instability—results from a dietary intake
and micronucleus index survey in South Australia. Carcinogenesis 26,
991–999, 2005.
129. Mazaki M, Ishii T, and Uyeta M: Mutagenicity of hydrolysates of citrus
fruit juices. Mutat Res 101, 283–291, 1982.
130. Ekasari I, Bonestroo MH, Jongen WMF, and Pilnik W: Mutagenicity and
possible occurrence of flavonol aglycones in heated orange juice. Food
Chem 31, 289–294, 1989.
131. Ekasari I, Berg HE, Jongen WMF, and Pilnik W: Characterization of
mutagenic compound(s) in heated orange juice. Food Chem 36, 11–18,
1990.
132. Friedman M, Wilson RE, and Ziderman II: Effect of heating on muta-
genicity of fruit juices in the Ames test. Journal of Agricultural and Food
Chemistry 38, 740–743, 1990.
133. Friedman M, Henika PR, Levin CE, and Mandrell RE:”Antibacterial ac-
tivities of plant essential oils and their components against Escherichia coli
O157 : H7 and Salmonella enterica in apple juice. Journal of Agricultural
and Food Chemistry 52, 6042–6048, 2004.
134. Sayre RM and Dowdy JC: The increase in melanoma: Are dietary furo-
coumarins responsible? Medical Hypotheses 70, 855–859, 2008.
135. Henriques JA and Moustacchi E: Interactions between mutations for sen-
sitivity to psoralen photoaddition (pso) and to radiation (rad) in Saccha-
romyces cerevisiae. J Bacteriol 148, 248–256, 1981.
136. World Cancer Research and the American Institute for Cancer Research:
Food, Nutrition, Physical Activity, and the Prevention of Cancer: A Global
Perspective. American Institute for Cancer Research, Washington, DC,
2007.
Downloaded by [UNISC] at 15:03 06 August 2013
... Orange juice (OJ), the fruit juice most consumed worldwide, contains a wide array of micronutrients and phytochemicals that have been attributed preventive effects against the onset of several noncommunicable diseases, such as cardiometabolic diseases, neurological disorders and some types of cancer [1][2][3][4]. The list of OJ bioactive compounds include vitamin C, (poly)phenols (mainly flavanones), carotenoids (xanthophylls, cryptoxanthins, carotenes), folate, and melatonin [5,6]. ...
... A total of 12 phenolic metabolites were quantified in plasma samples collected over an 8 h period following ingestion of OJ or FOJ. The (poly)phenol metabolites were mainly phase II metabolites of flavanones (naringenin-7-glucuronide [compound 16], naringenin-4ʹglucuronide [17], hesperetin-7-glucuronide [19], hesperetin-3ʹ-glucuronide [20], hesperetin-diglucuronide [23], and hesperetindiglucuronide II [24]), although three phenolic acids (3ʹ-methoxycinnamic acid-4ʹ-glycine [8], 3ʹ-methoxycinnamic acid-4ʹ-sulfate [9], and 3-(4ʹ-methoxyphenyl)propanoic acid-3ʹ-glucuronide [14]) as well as three hippurates (hippuric acid [3], 4ʹ-hydroxyhippuric acid [4], and 3ʹhydroxyhippuric acid [5]) were also quantified. Hippuric acid derivatives were not included in the calculation of the total amount of (poly)phenolic metabolites since they also originate from other dietary and endogenous sources [39]. ...
Article
The consumption of orange juice provides high concentrations of health-promoting bioactive compounds, the amount of which may increase upon alcoholic fermentation. Although fermentation may offer new prospects for the industry of orange-related products, there is a lack of studies reporting the influence of controlled alcoholic fermentation on the bioavailability of orange juice (poly)phenols in humans. The aim of this study was to evaluate the absorption profile, pharmacokinetic parameters, and urinary excretion of orange juice (poly)phenols in nine volunteers after acute administration of an orange juice and a beverage prepared after controlled alcoholic fermentation of the juice. Plasma and urine samples were analysed through a UHPLC-ESI-MS/MS targeted approach. A total of 24 (poly)phenol metabolites including both flavanone and phenolic acid derivatives were quantified, most of them being recorded only in urine. Phase II conjugates of hesperetin and naringenin were the main metabolites in plasma, while phenolic acids, in particular hydroxybenzoic acids, were the main compounds in urine. (Poly)phenols in both beverages were highly bioavailable (between 46 and 59%) and a notable inter-individual variability was seen. Significant treatment × time interactions were recorded for the sum of flavanones and phenolic acids in plasma, the (poly)phenols in the fermented juice being absorbed faster than after orange juice intake. Nevertheless, despite the food matrix having an impact on the absorption profile of orange juice (poly)phenols, this did not influence the pharmacokinetic parameters and urinary excretion of the (poly)phenol metabolites.
... New strategies for suppressing cancer cells with bioactive compounds from food have been investigated, for instance the suppression of the nuclear factor kappa B (NF-κB), a transcription factor linked to carcinogenesis (Barthi and Aggarwal, 2002). Chemoprevention is a promising and rational approach that uses synthetic, natural, or biological agents to reduce or block the occurrence of cancer (Franke et al., 2013;Steward and Brown, 2013). In line with this perspective, epidemiological studies have shown that the consumption in long-term of balanced diets, particularly those rich in vegetables and fruits, can promote chemopreventive activities (Steinmetz and Potter, 1996;Meiyanto et al., 2012). ...
... Indeed, the beneficial effects of these diets are attributable, at least in part, to polyphenols (Ullah and Khan, 2008;Singh et al., 2012juices, are an abundant source of citrus flavonoids, a subclass of polyphenols that represent an important dietary component. Inverse correlations have been reported between the consumption of orange juice and the incidence of cancer (So et al., 1996;Miyagi et al., 2000;Tanaka et al., 2012;Franke et al., 2013;Farooqi et al., 2015). The anticancer properties of orange juice have been focused on hesperidin and naringin, flavanones found almost exclusively in citrus (Guthrie et al., 1998;Ghorbani et al., 2012;Zeng et al., 2014). ...
Article
Full-text available
Background: Red-fleshed sweet orange juice (ROJ) comes from a new variety of citrus cultivated in Brazil that contains high levels of β-carotene and lycopene, and similar amounts of hesperidin (HSP) and nutrients, equivalently to blond orange juice (BOJ). Such bioactive compounds are associated with chemopreventive actions in several cancer cell lines. The purpose of this study was to examine the cytotoxicity, cell cycle, apoptosis, and cytokine secretion after BOJ, ROJ, and HSP treatment of a novel T acute lymphoblastic leukemia cell line, Loucy. Materials and methods: Loucy cells were incubated for 24-h with BOJ, ROJ, and HSP, and the viability was measured using trypan blue. Cell cycling and apoptosis were assessed by propidium iodide (PI) and annexin V-FITC/PI flow cytometry, respectively. Secretion of cytokines IL-1α, IL1-β, IL-2, IL-4, IL-6, IL-10, IL-17A, IFNγ, TNFα, TGFβ, MIPα, and MIPβ was determined by ELISA array. Results: BOJ and ROJ treatments promoted Loucy cell cytotoxicity. Additionally, BOJ induced cell cycle arrest in the G0/G1 phase, and decreased the cell accumulation in the G2/M. ROJ decreased only the G0/G1 fraction, while HSP did not change the cell cycle. BOJ led to apoptosis in a different fashion of ROJ, while the first treatment induced apoptosis by increase of late apoptosis and primary necrotic fractions, the second increased early and late apoptosis, and primary necrotic fraction compared to positive controls. HSP had no effect on apoptosis. IL-6 and IL-10 were abrogated by all treatments. Conclusions: Taking together, these results suggest potential chemopreventive effects of BOJ and ROJ on Loucy cells.
... Among fruit juices, orange juice (OJ) is the most consumed worldwide and several intervention studies suggests that its regular consumption could provide additional protective activity against the onset and progression of several chronic diseases, such as CVD and some types of cancer [14,15]. These positive effects have been attributed to its profile of bioactive compounds such as vitamin C, (poly)phenols, carotenoids, folate, and melatonin [16][17][18] which exert a large number of biological activities: antioxidant activity, regulation of lipid profile, improvement of endothelial function, anti-inflammatory function, anti-cancer property, glycemic regulation, improvement of immune function, and reproductive and bone metabolism [19][20][21][22][23][24][25][26][27][28]. ...
Article
Higher postprandial plasma glucose and lipemia, and oxidative and inflammatory responses, are considered important cardiovascular risk factors. Fermentation of fruits has generated products with high concentrations of bioactive compounds. The aim of this study was to evaluate the potential acute effects that fermented orange juice (FOJ) can exert in healthy humans by modulating postprandial response, and inflammatory/antioxidant status, compared with orange juice (OJ). Nine volunteers were recruited for a randomized, controlled, and crossover study. Participants ingested 500 mL of FOJ. At 4 h post intake, subjects consumed a standardized mixed meal. Blood samples were collected at 0-8 h hours post intake. The subjects repeated the protocol with OJ following a 2-week washout period. Glucose and lipid metabolism, plasma antioxidant capacity (ORAC, FRAP), endogenous antioxidants (albumin, bilirubin, uric acid), C-reactive protein and fibrinogen were measured in plasma samples. There was a trend of a smaller increase in LDL-C after FOJ intake compared with OJ, a significant decrease in apo-B and significant increase in ORAC. The glycemic and triglyceride response of meal was attenuated with FOJ. No differences were obtained in endogenous antioxidants and inflammation status between the treatments. The acute consumption of FOJ could play a protective role against cardiovascular risk factors.
... Around more than 3000 plants have been recorded to possess ant cancerous characteristics; therefore, these medicinal plants are used as an alternative treatment strategy in different countries. This alternative approach to prevent or delay cancer development is called chemoprevention, which utilizes medicinal herbs and food [23][24][25] . Among such medicinal plants include Kola nuts of the Sterculiaceae family of plants. ...
Article
Full-text available
Kola nut extracts have recently been reported to contain chemopreventive compounds providing several pharmacological benefits. This study investigated Kola nut extracts' anti-cancer activity on human immortalized myelogenous leukemia cell line K562 through apoptosis and cell cycle arrest. Fresh Kola nuts were prepared as powder and dissolved in DMSO. Different concentrations (50, 100, 150, 200, and 250 μg/ml) of working solutions were prepared. The K562 cells were treated with the different concentrations of Kola nut extract or vehicle control (10% DMSO) followed by incubation at 37°C for 24, 48, and 72 hours, respectively. Treatment activity was investigated in K562 cells; by Resazurin, and FITC/Propidium Iodide and 7-AAD stained cells to evaluate apoptotic cells and the cell cycle's progression. Inhibition of leukemia cell proliferation was observed. The extract effectively induced cell death, early and late apoptosis by approximately 30% after 24 and 48 hours incubation, and an increase in the rate of dead cells by 50% was observed after 72 hours of incubation. Also, cell growth reduction was seen at high dose concentrations (150 and 200 µg/ml), as evident by cell count once treated with Kola nut extract. The total number of apoptotic cells increased from 5.8% of the control group to 27.4% at 250 µg/ml concentration. Moreover, Kola nut extracts' effects on K562 cells increased gradually in a dose and time-dependent manner. It was observed that Kola nut extracts could arrest the cell cycle in the G2/M phase as an increase in the number of cells by 29.8% and 14.6 % were observed from 9.8% and 5.2% after 24 and 48 hours of incubation, respectively. This increase was detected in a dose and time-dependent manner. Kola nut extracts can be used as a novel anti-cancer agent in Leukemia treatment as it has shown significant therapeutic potential and therefore provides new insights in understanding the mechanisms of its action. Keywords: Kola nut extracts, Leukemia, K562 cell line, Apoptosis, Cancer.
... They summarized the consumer behaviour of three kinds of orange juice in the main market worldwide [9]. A large number of studies have shown that eating a large amount of citrus fruits is beneficial to anti-cancer functions and prevention of cardiovascular diseases [10][11][12][13]. ...
Article
Full-text available
Boba milk tea is very popular around the world. The “boba” balls in milk tea are usually made of tapioca. Reports on calcium alginate ball encapsulation in fruit-flavoured drinks have rarely been seen. The preparation method for this kind ball was studied. The “boba” balls were obtained by membrane formation on the interface through the addition of calcium chloride fluids into a sodium alginate solution. The operation conditions were studied, including drop height, flow velocity, sodium alginate and calcium chloride solution concentration. The diameter, mechanical strength, loading ratio and encapsulation rate of the “boba” balls are discussed. The optimized preparation conditions were as follows: the diameter of adding tube was 8 mm, the drop height was 25 cm, the drop flow rate was 60 mL/min, 1.0% sodium alginate, 1.0% calcium chloride. The prepared “boba” balls were stored at different temperatures. No microorganisms were detected in 90 days, and the sensory quality decreased with storage time. Shelf life was predicted using the Arrhenius equation; when the storage temperature was less than 10 °C, it could be stored for more than 1 year. This preparation technology of “boba” balls has potential for application by milk tea ingredient companies or relevant beverage manufacturing factories.
... More recently, putresine, spermidinem, spermine, and TRY have been also reported in the orange fruit. Some studies have reported the chemopreventive properties of orange juice associated with its effect on metabolic enzymes and its anti-inflammatory, cytoprotective/apoptotic, hormonal, cell signaling-modulating, antioxidant, and antigenotoxic effects (Franke et al. 2013). Also, orange juice provides several kinds of minerals and vitamins necessary to healthy state (Lee et al. 2014). ...
Article
Full-text available
Recent reports have shown that commercial orange juice is rich in biogenic amines. Consumption of foods containing large amounts of biogenic amines increase hypertensive crisis and high levels of histamine and tyramine, which have been implicated as causative agents in a number of food poisoning episodes. In addition, accumulation of tryptamine in plasma may be associated with mood disorders. The aim of this study was to determine whether chronic administration of orange juice extract and tryptamine affects the behavior and c-fos expression in the rat. For this purpose, Wistar male rats were injected with saline solution, tryptamine or orange juice extract. Sucrose preference test and elevated plus maze were evaluated to determine hedonic and anxiety behavior, respectively. Rats treated with orange juice extract showed increased anxiety behavior and sucrose consumption, similar to those treated with tryptamine. In addition, dorsal raphe nucleus, accumbens nucleus, and hippocampus showed an increase of c-fos positive cells in rats treated with orange juice extract. In conclusion, the chronic and lengthy consumption of orange juice or their derivatives in the diet could be a factor responsible to induce mood disorders and may promote excess caloric consumption.
... [7][8][9] Furthermore, numerous epidemiological and intervention studies have demonstrated the protective and therapeutic role of regular orange juice consumption against chronic diseases, including cardiovascular disorders, diabetes, obesity and cancer. 3,[10][11][12] Observations of these beneficial effects of orange juice have increased research interest in technological treatments to preserve or enhance the bioactive compounds it contains. 13,14 Alcoholic fermentation of various types of fruit 15,16 has generated new products with moderate alcohol content and a higher concentration of bioactive compounds than in the original substrate. ...
Article
Background: Alcoholic fermentation of fruits has generated novel products with high concentrations of bioactive compounds and moderate alcohol content. The aim of this study was to evaluate the potential effect on cardiovascular risk factors of the regular consumption by healthy humans of a beverage obtained by alcoholic fermentation and pasteurization of orange juice. Results: Thirty healthy volunteers were enrolled in a randomized controlled study. The experimental group (n=15) drank 500 mL orange beverage (OB) day(-1) for 2 weeks (intervention phase), followed by a 3-week washout phase. Blood samples were collected at baseline (E-T0) and at the end of the intervention (E-T1) and washout (E-T2) phases. Controls (n=15) did not consume OB during a 2-week period. OB intake significantly increased ORAC (43.9%) and reduced uric acid (-8.9%), CAT (-23.2%), TBARS (-30.2%), and C-reactive protein (-2.1%) (E-T1 vs. E-T0). These effects may represent longer-term benefits, given the decreased uric acid (-8.9%), CAT (-34.6%), TBARS (-48.4%), and oxidized LDL (-23.9%) values recorded after the washout phase (E-T2 vs. E-T0). Conclusion: The regular consumption of OB improved antioxidant status and decreased inflammation state, lipid peroxidation, and uric acid levels. Thus, OB may protect the cardiovascular system in healthy humans and be considered a novel functional beverage.
Article
The aim of this study was to develop a monoclonal antibody (mAb)-based enzyme-linked immunosorbent assay (ELISA) for the quantification of a major allergen (Cit s 2) in fresh and processed oranges. Purified recombinant Cit s 2 (rCit s 2)-small ubiquitin-like modifier (SUMO) was used for the production of mAbs. In the optimized ELISA, the recovery of rCit s 2 from Navel oranges or orange juice was 107–132%, and the intra- and inter-assay coefficients of variation were 3.1–8.8% and 4.4–11%, respectively. The Cit s 2 content in fresh oranges was determined to be 1,800 ± 430 ng/g, while this content was much lower in the processed foods. The developed ELISA demonstrated high reproducibility, sensitivity, and accuracy, and this assay may help individuals with orange allergy by determining Cit s 2 quantities in food products and controlling their Cit s 2 intake.
Chapter
The health benefits of fruits are believed to be due, in large part, to phytochemicals and among them polyphenols. Citrus fruit and juices are rich in flavonoids, the two more prominent being hesperidin in oranges and naringin in grapefruit. The most beneficial effects of citrus phytochemicals seem to be generated by dampening of chronic inflammation. This slow debilitating process, sometimes referred to as “the silent killer,” is considered to be at the root of most chronic diseases, including cardiovascular diseases (CVD), type 2 diabetes, osteoporosis, dementia, and some forms of cancer. Citrus juices have been reported by epidemiological, in vitro, animal, and clinical studies to have a beneficial influence on most of those diseases. The most convincing clinical evidence is associated with reduction of the risk of development of CVD and improving cognitive activities and function. However, the benefits are mild and without the more dramatic effects expected with drug therapies. This review examines the beneficial health associations between citrus consumption and chronic diseases, particularly in the future area of epigenetics.
Article
Orange juice is a rich source of bioactive compounds. Fermentation processes have been carried out in fruits, resulting in products with higher bioactive compound contents than the substrates. The aim of this study was to evaluate changes in phenolic acids, flavones and flavanone derivatives during the alcoholic fermentation process (15 days) in orange juice and to optimize the fermentation time. A total of 45 (poly)phenolic compounds were detected by UHPLC coupled with a linear trap quadrupole (LTQ) and Orbitrap Elite series mass analyser (UHPLC-Orbitrap-MS/MS). We tentatively identified 21 hydroxycinnamic acids, including ferulic acid, caffeic acid, and sinapic acid, in addition to 18 hydroxycinnamic acid derivatives (7 ferulic acid derivatives, 8 caffeic acid derivatives, 2 sinapic acid derivatives, a p-coumaric acid derivative) as well as 2 hydroxybenzoic acid derivatives, a hydroxypropionic acid derivative and other compounds (citric acid, quinic acid, 3 quinic acid derivatives) for the first time in fermented orange juice. In addition, 16 flavonoids, 7 flavanones (didymin, hesperidin, narirutin and 4 narirutin derivatives), 7 flavonols (kaempferol derivatives) and 2 flavones (diosmetin, vicenin-2) were putatively identified in fermented orange juice for the first time. Total hydroxycinnamic acid, benzoic acid, flavones and flavonol derivative contents showed significant increases (7.9, 4.7, 18.3 and 24.5%, respectively) on day 11 of fermentation relative to the original juice. The optimum time for the procedure was 11 days, after which the highest content of (poly)phenolic compounds was reached. The potential beverage produced by alcoholic fermentation of orange juice would exert greater health effects in humans than the substrate, derived from both the (poly)phenolic content and the low level of alcoholic content.
Article
Full-text available
Authors' conclusions The failure of vitamin C supplementation to reduce the incidence of colds in the general population indicates that routine vitamin C supplementation is not justified, yet vitamin C may be useful for people exposed to brief periods of severe physical exercise. Regular supplementation trials have shown that vitamin C reduces the duration of colds, but this was not replicated in the few therapeutic trials that have been carried out. Nevertheless, given the consistent effect of vitamin C on the duration and severity of colds in the regular supplementation studies, and the low cost and safety, it may be worthwhile for common cold patients to test on an individual basis whether therapeutic vitamin C is beneficial for them. Further therapeutic RCTs are warranted. AVAILABLE AT: http://www.mv.helsinki.fi/home/hemila/CC/CC.htm
Article
Full-text available
Juices from the mandarin Clemenules (Citrus clementina Hort. ex Tan.), the tangor Ortanique (Citrus reticulata Blanco × Citrus sinensis Osb.) and the sweet orange Valencia Late (Citrus sinensis) have been industrially squeezed, pasteurized, concentrated and stored under refrigeration (4 °C) and at room temperature (20 °C). After each process, the flavanone-7-O-glycosides (FGs) and fully methoxylated flavones (FMFs) contents as well as total, cumulative fast-kinetics and cumulative slow-kinetics antiradical activities were determined and compared with those from the corresponding fresh hand-squeezed juices. Neither industrial-squeezing, nor pasteurization or concentration significantly affected FGs and FMFs contents and antiradical activities of assayed juices. Storage caused a slight decrease of the FMFs contents but a significant reduction of both soluble hesperidin contents and cumulative fast-kinetics antiradical activities in all assayed juices. These decreases were dependent on storage temperature. Characteristic values of the varietal characterization parameters, which are derived from the FMFs contents and antiradical activities of fresh hand-squeezed juices, held valid for industrially squeezed, pasteurized and concentrated juices. After storage, however, only the FMFs-derived varietal characterization parameters and cumulative slow-kinetics antiradical activity remained valid for the resulting juices.
Article
We examined the bioavailability of vitamin C in orange juice processed using high pressure (HP) and its effects on plasma levels of vitamin C, uric acid (UA), F2-isoprostanes (8-epiPGF(2alpha)), C-reactive protein (CRP) and prostaglandin E-2 (PGE(2)) in a healthy human population. Subjects (6 men, 6 women) enrolled in the study consumed 500 mL/d of HP orange juice for 14 cl, corresponding to an intake of 250 mg of vitamin C. On d 1 of the study, subjects drank the juice in one dose; on d 2 until the end of the study, d 14, they drank 250 mL in the morning and 250 mL in the afternoon. Blood was collected every h for 6 h, on d 1, and then on d 7 and 14 of the study. Baseline plasma vitamin C concentration was higher (P = 0.014) in women (55.8 +/- 3.8 mumol/L) than in men (42.8 +/- 2.1 mumol/L). The maximum plasma vitamin C increase occurred 3 h after drinking the juice, and it remained elevated on d 7 and 14. Plasma 8-ep/PGF(2alpha) concentration did not differ between men and women at baseline. However, it was lower at the end of the study in both men (P = 0.044) and women (P = 0.034). Plasma levels of vitamin C and 8-ep/PGF(2alpha) were inversely correlated (r = -0.615, P = 0.001). Plasma CRP concentrations tended to be lower on d 14 than at baseline in men (P = 0.317) and women (P = 0.235). Plasma PGE(2) was lower at the end of the study in both men and women (P less than or equal to 0.037). Drinking orange juice increases plasma vitamin C, and decreases 8-ep/PGF(2alpha) and PGE(2) levels in humans, which may help reduce the risk of chronic diseases.
Article
Collectively, the evidence from epidemiologic, animal and human studies strongly suggests that folate status modulates the risk of developing cancers in selected tissues, the most notable of which is the colorectum. Folate depletion appears to enhance carcinogenesis whereas folate supplementation above what is presently considered to be the basal requirement appears to convey a protective effect. The means by which this modulation of cancer risk is mediated is not known with certainty, but there are several plausible mechanisms which have been described. Folate plays a major role in the formation of S-adenosylmethionine, the universal methyl donor, as well as in the formation of purine and thymidine synthesis for DNA and RNA. Therefore, most mechanistic studies performed to date have focused on alterations in DNA methylation, disruption of DNA integrity and disruption of DNA repair, all of which have been observed with folate depletion. These aberrations in DNA are believed to enhance carcinogenesis by altering the expression of critical tumor suppressor genes and proto-oncogenes. Recently, the role of a common polymorphism of the methylenetetrahydrofolate reductase gene has been highlighted as well. This review presents those mechanisms which are the most likely candidates to explain folate's effects and it proposes an integrated scheme to explain how these mechanisms might interact.
Article
The objective of the present study was to examine and compare the effects of frequently consumed beverages on the human intestinal cell line, Caco-2, in terms of toxicity, growth, and differentiation. For this purpose, Caco-2 cells were incubated for 24 h in the presence of: a mineral water, fresh orange juice, packaged orange juice, a cola drink, an energy drink, black, camomille, and green teas, and drip coffee. Toxicity was evaluated firstly by measuring the lactate dehydrogenase leakage from the cells and secondly by performing the MTT assay. Cell differentiation was determined by measuring two brush border membrane enzyme activities. Growth rates of Caco-2 cells were evaluated by cell counting. The results indicated that there were no significant differences between the beverages in toxicity and cell proliferation. Cells exposed to fresh orange juice exhibited higher tetrazolium reduction rates in the MTT assay (121.3% of control). These cells also showed higher succinate cytochrome c reductase activities than the other samples, implying that the contents of fresh orange juice, such as ascorbic acid, stimulated mitochondrial metabolism. The alkaline phosphatase and aminopeptidase N activities of the Caco-2 cells lay between 61.50 and 110.00%, indicating a partial influence of some beverages on Caco-2 differentiation.