Crude Polysaccharides from Okra Pods (Abelmoschus esculentus)
Grown in Indonesia Enhance the Immune Response due to
Sri Puji Astuti Wahyuningsih , Manikya Pramudya, Intan Permata Putri, Dwi Winarni,
Nadyatul Ilma Indah Savira, and Win Darmanto
Department of Biology, Faculty of Science and Technology, Airlangga University, Surabaya 60115, Indonesia
Correspondence should be addressed to Sri Puji Astuti Wahyuningsih; firstname.lastname@example.org
Received 23 March 2018; Revised 24 August 2018; Accepted 19 September 2018; Published 9 October 2018
Academic Editor: Paola Patrignani
Copyright ©2018 Sri Puji Astuti Wahyuningsih et al. is is an open access article distributed under the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
Okra pods were widely consumed by Indonesians to maintain health. e aim of this study was at investigating the potential of
crude polysaccharides from okra pods on immune response in mice infected with Staphylococcus aureus. irty male Balb/C
mice were divided into six groups: normal control, negative control, and treatment groups (administration of crude poly-
saccharides at doses of 25, 50, 75, and 100 mg/kg). Crude polysaccharides were administrated for fourteen days. Furthermore,
mice were exposed to S. aureus at the ﬁfteenth day. Two weeks after the end of treatment, the parameters were measured. is
study showed that crude polysaccharides at a dose of 75 and 100 mg/kg improved phagocytic activity, spleen index, and
splenocytes proliferation. Rising of TNF-αlevels was shown in groups treated with crude polysaccharides at doses of 25, 50, and
100 mg/kg. All treatment groups showed a decreasing level of IL-17. Crude okra polysaccharides also showed a slight increase
in NK cells activity and IFN-clevel. us, crude okra polysaccharides could act as an eﬀective material to enhance immune
response including phagocytic activity, spleen index, splenocytes proliferation, and control immune responses through
e human body is surrounded by environment-contained
microbes, including extracellular bacteria, S. aureus. ese
bacteria are able to cause nosocomial infection which can
result in serious infections . Normally, immune-related
cells will inhibit S. aureus transmission but the bacteria also
release components against the immune system. erefore,
the body needs a particular compound to enhance the
Dietary phytochemicals from plants may play important
roles in the prevention of many diseases . Plant poly-
saccharides have been known as an important immunos-
timulatory agent with broad spectrum, low toxicity, and few
side eﬀects . If polysaccharides are component of our
daily food, it will give many health beneﬁts for human body.
Okra (A. esculentus) is vegetable crop used as food and
traditional medicine for many diseases such as dysentery and
diarrhea [4, 5]. Okra contains ﬂavonoid and vitamin C as
antioxidant and polysaccharides as an immunomodulator
[6, 7]. A study with cyclophosphamide as an antigen has
revealed that okra polysaccharides increased spleen index,
splenocyte proliferation, and cytokines secretion . e
extract of okra increased IL-12 secretion and decreased IL-10
secretion in dendritic cells . Related with bacterial in-
fection, okra fruit has high tannins that could abolish
However, studies have not been reported on the po-
tential of crude polysaccharides from okra pods consumed
in Indonesia to overcome high cases of S. aureus infection.
To further investigate the potential of crude okra poly-
saccharides, the present study explored the eﬀect by
Advances in Pharmacological Sciences
Volume 2018, Article ID 8505383, 7 pages
evaluating phagocytic activity, cytokine production, spleen
index, splenocytes proliferation, and NK cell activity.
2. Materials and Methods
2.1. Materials and Chemicals. Okra pods were collected from
the traditional market in Surabaya, Indonesia, in May 2017.
e okra pods were packaged 500 g per polyethylene bag and
then stored at −20°C until use. S. aureus (ATCC 25923) was
purchased from Balai Besar Laboratorium Kesehatan, Sur-
abaya, Indonesia. RPMI-1640 was purchased from Gibco
(Invitrogen Co, Massachusetts, USA). Lipopolysaccharide
(LPS) from Escherichia coli 055 : B5, L2880, and lyophilized
powder were purchased from Listlab (List Biological
Labs, Inc., California, USA). 3-(4, 5-dimethylthiazol-2-yl)-2,
5-diphenyltetrazolium bromide (MTT) and human hepa-
toma cell line (huh7it) were acquired from Institute of
Tropical Disease, Airlangga University (Surabaya,
Indonesia). Interleukin-17 (IL-17), interleukin-23 (IL-23),
interferon-c(IFN-c), and tumor necrosis factor (TNF)-α,
enzyme-linked immunosorbent assay (ELISA) kit were
purchased from BioLegend (BioLegend, Inc., San Diego,
USA). All other chemicals and solvent used were of ana-
lytical reagent grade.
2.2. Preparation of Crude Polysaccharides from Okra Pods.
According to Ramesh et al.  and Chen et al. , frozen
okra pods (500 g) were cleaned with distilled water, cut into
small slices, homogenized, and macerated with 500 ml
double-distilled water (ddH2O) overnight. e extract of
okra was ﬁltered and macerated twice again with 300 ml
ddH2O. e supernatants were collected by centrifugation at
4300 rpm for 5 min. e supernatants were precipitated by
the addition of anhydrous ethanol 1X sample volume and
incubated for 24 h at 4°C and then centrifuged again as
above. e precipitated material was then dissolved in
O and dialyzed through cellulose membrane (Sigma-
Aldrich, retaining >Mw 14,000) for 24 h. e aqueous
solution was then collected from the dialysis bag and freeze-
dried to obtain the crude okra polysaccharides.
2.3. Determination of Polysaccharides Content in Okra Pods.
Polysaccharides content in okra pods was determined using
phenol sulphuric acid assay. Sample solution of crude okra
polysaccharides was made from stock of crude okra poly-
saccharides (10 µL) and aquadest (90 µL). en, 50 µL of
phenol 5% was added. After being homogenized for 1 min,
2 mL of sulphuric acid was added to the solution and in-
cubated for 10 min in room temperature. e blank solution
was made from 50 µL of phenol 5% and 100 µL of aquadest.
e absorbance was measured at 490 nm.
2.4. Animals. Male BALB/c mice (8–10 weeks old, 30–40 g)
were provided by the Laboratory Animal from Faculty of
Pharmacy, Airlangga University, Surabaya, Indonesia. e
animals were maintained in cages made of a plastic with a lid
made of woven wire cage at 20°C, with 12 h light/12 h dark
cycle, fed and watered by ad libitum. All procedures
involving animal care were approved by Animal Care and
Use Committee (ACUC) of Veterinary Faculty, Airlangga
University, Surabaya, Indonesia, no. 714-KE.
2.5. Experimental Design. After 10 days of acclimatization,
BALB/c mice were randomly divided into six groups (KN:
normal control without any treatment; K−: negative control
exposed by S. aureus without okra crude polysaccharides
administration; P1, P2, P3: okra crude polysaccharides doses
25, 50, 75, and 100 mg/kg BW, respectively). Okra crude
polysaccharides were administrated by gavage in fourteen
days. Furthermore, mice were exposed to S. aureus (0.5 Mc.
Farland) once through intraperitoneal at the ﬁfteenth days.
Two weeks after the last administration, the animals were
weighed, blood samples were collected to obtain serum, and
then the animals were killed. e intraperitoneal ﬂuid was
collected. e spleen was excised from the animal and
weighed immediately and placed in cold PBS-penicillin-
streptomycin. e relative was calculated according to the
following formula: spleen index (mg/g) �(weight of
2.6. Phagocytic Activity Assay. e mice were injected in-
traperitoneally with 0.2 mL of S. aureus suspension. One
hour later, the mice were killed by ketamine anesthesia, and
3 mL of 3% EDTA was used as an anticoagulant. en, the
intraperitoneal ﬂuid was collected. Intraperitoneal suspen-
sion was smeared on glass slides and air-dried. e smear
was ﬁxed using methanol for 15 minutes and stained with
Giemsa solution for 20 mins. Phagocytic activity was de-
termined by counting the number of phagocytes in a pop-
ulation of 100 phagocytes.
2.7. Serum Cytokine Assay. Whole blood was collected and
centrifuged at 3000 rpm and 4°C for 10 min, while the upper
layer contained the serum. e levels of IFN-c, TNF-α, IL-
17, and IL-23 in the serum were analyzed by commercial
enzyme-linked immunosorbent assay (ELISA) kits (BioL-
egend, Massachusetts, USA) according to the manufacturer’s
protocol. e absorbance was measured using ELISA reader
at 450 nm.
2.8. Splenocytes Isolation. e spleens were gently homog-
enized and passed through a sterilized copper sieve (200-
mesh) to obtain single cell suspensions. Splenocytes sus-
pension was centrifuged at 1000 rpm for 5 minutes. Pellet
containing red blood cells was resuspended in tris-buﬀered
NH4Cl pH 7.2 and centrifuged at 1000 rpm for 5 minutes
until white pellet was obtained. Splenocytes were washed
with 5 ml of PBS-100 unit/ml penicillin-100 µg/ml strepto-
mycin and resuspended in RPMI 1640-FBS 10% medium.
en, splenocytes were used in splenocytes proliferation
assay and natural killer cell activity assay.
2.9. Splenocytes Proliferation Assay. Cell numbers of sple-
nocytes were counted by haemocytometer. 195 µl of
2Advances in Pharmacological Sciences
splenocytes (3 ×105 cells/well) and 5 µl of LPS (200 µg/ml)
were seeded in 96-well plates. After incubation at 37°C in an
incubator with 5% CO
for 48 hours, MTT assay was used.
e absorbance was measured at 560 nm. Splenocytes
proliferation (%) �((OD value of okra-treated cells)/(OD
value of control)) ×100.
2.10. Natural Killer Cell Activity Assay. e splenocytes as
the eﬀector (3 ×105 cells/well) was added to human he-
patocyte cell line as the target cells (6 ×103 cells/well) to give
E/T ratio 50 : 1. ey were cultured in 96-well plate and
incubated at 37°C in an incubator with 5% CO
for 48 hours.
e activity of NK cell was evaluated by the MTT assay. e
absorbance was measured at 560 nm. NK cell activity (%) �
((OD value of okra-treated cells)/(OD value of control)) ×
2.11. Statistical Analysis. Statistical analysis was performed
by one-way analysis of variance (ANOVA) followed by
Duncan’s post hoc test. All analysis was performed using
IBM SPSS Statistics 24 software. e results were reported as
the mean ±standard deviation (SD) of ﬁve repeats. Pvalue
of <0.05 was considered statistically signiﬁcant.
3.1. Determination of Polysaccharide Content in Okra Pods.
Using the polysaccharide standard regression equation, the
polysaccharides content in the stock solution with dose of
100 mg/kg BW was 22.87 mg/mL.
3.2. Phagocytic Activity. Phagocytic activity was signiﬁcantly
increased in P3 group and P4 group compared to normal
control group and negative control group (P<0.05). e
highest increase on phagocytic activity was shown by P3
group. Meanwhile, P1 group and P2 group increased
phagocytic activity but did not show signiﬁcant diﬀerence
compared with normal and negative control groups
3.3. Cytokines Production. Serum levels of TNF-αwere
signiﬁcantly increased in P1 group (423.20 ±128.66 pg/ml),
P2 group (460.40 ±79.28 pg/ml), and P4 group (282.40 ±
80.38 pg/ml) compared to normal control and negative
control groups (P<0.05). P3 group (175.50 ±79.76) also
showed slight increase of TNF-αlevel compared to normal
control and negative control groups (P>0.05) (Table 1).
In contrast, IL-17 level in P2, P3, and P4 groups was
signiﬁcantly lower than normal control and negative control
groups (P<0.05). Although the diﬀerence was not signiﬁ-
cance, P1 showed decrease level of IL-17. ere was slight
increase in the serum level of IFN-cin P4 group (174.60 ±
64.55 pg/ml) compared to normal control and negative
control groups but did not show a signiﬁcant diﬀerence.
Meanwhile, there was no diﬀerence in the result of IL-23
3.4. Spleen Index. Spleen index was signiﬁcantly increased in
P2 group (14.84 ±2.76%) and P3 group (14.43 ±3.31%)
compared to normal control group (P<0.05). P4 group at
a dose of 100 mg/kg showed highest spleen index (16.72 ±
3.60%) and increased spleen index signiﬁcantly compared to
normal control and negative control groups (P<0.05)
3.5. Splenocytes Proliferation. Splenocytes proliferation was
signiﬁcantly increased in the P3 group and P4 group
compared with other groups (P<0.05). e highest increase
of splenocytes proliferation was demonstrated in P3 group
with 157.77 ±11.06% (Figure 3).
3.6. Natural Killer Cell Activity. K−group and all the
treatment groups (P1, P2, P3, and P4) did not show sig-
niﬁcant increase of natural killer cell activity compared with
normal control group. P3 and P4 groups showed slight
increase in NK cell activity (104.67 ±15.32% and 106.75 ±
15.32%) compared to normal control, which was similar to
the results of the splenocytes proliferation (Figure 4).
Immune system gives protection to organism against bac-
terial infection through layered defense, nonspeciﬁc im-
munity, and speciﬁc immunity. When bacteria pass through
nonspeciﬁc defense, the body forms a more complex im-
mune system . ese days, the use of immunomodu-
lators to improve immune responses has been considered
one of the promising alternatives to prevent bacterial in-
fection . One of the potential compounds as immuno-
modulator is polysaccharide. In this study, we used crude
polysaccharides which contain 22.87 mg/mL of poly-
saccharides after phenolic sulphuric acid assay was
Nonspeciﬁc immunity component that ﬁrstly recognizes
bacteria is macrophage . Activation of macrophages
KN K– P1 P2 P3 P4
Phagocytic activity (%)
Figure 1: Eﬀect of crude polysaccharide from okra pods on
phagocytic activity (%). KN: normal control; K−: negative control;
P1, P2, P3, and P4 were treated with 25, 50, 75, and 100 mg/kg BW
crude okra polysaccharides, respectively. Each bar represents
means ±SD (n�5). ∗∗P<0.05 compared to normal control.
Advances in Pharmacological Sciences 3
plays a key role in nonspeciﬁc immunity for developing
a defensive reaction against pathogens via phagocytosis
process . Macrophages release products such as oxygen
radicals and tumor necrosis factor that are harmful to
Polysaccharides regulate the host immune system by
activating immune cell-related to lymphocytes, macro-
phages, and NK cells . e present result demonstrated
that crude polysaccharides from okra pods at doses of
75 mg/kg and 100 mg/kg signiﬁcantly increased phagocytic
activity of intraperitoneal phagocytes. Previous observations
also demonstrated phagocytes activation by polysaccharides
from okra pods both in vitro and in vivo [3, 7]. It has been
reported that crude okra polysaccharides signiﬁcantly in-
creased NO production on RAW264.7 macrophage . In
this study, the rise of active phagocytes may lead to increase
of proinﬂammation cytokines production such as TNF-α,
IFN-c, IL-17, and IL-23 and improves host defense against
bacteria. TNF-αand IFN-care cytokines produced by
macrophage and cell due to all kinds of antigen in-
fections. Macrophage also produced IL-23 to induce acti-
vation of 17. Meanwhile, IL-17 is produced by 17
speciﬁcally. Staphylococcus aureus is extracellular bacteria.
Extracellular bacteria speciﬁcally induce na¨
ıve to dif-
ferentiate as 17 . Based on this consideration, we
examine level of TNF-α, IFN-c, IL-17, and IL-23.
Cytokines are small proteins produced by cells such as T
helper, NK cells, and macrophages that regulate the immune
Table 1: Eﬀect of crude polysaccharide from okra pods on cytokines production (pg/ml).
Groups Cytokine (pg/mL)
KN 116.70 ±78.23 113.00 ±39.78 208.50 ±85.67 13.348 ±0.11
K−157.59 ±41.95 156.80 ±51.00 204.50 ±84.65 13.300 ±0.01
P1 423.20 ±128.66∗∗∗ 98.40 ±50.98 127.50 ±78.77 13.377 ±0.04
P2 460.40 ±79.28∗∗∗ 139.20 ±23.38 73.25 ±22.53∗∗ 13.319 ±0.03
P3 175.50 ±79.76 97.20 ±41.44 78.25 ±50.42∗∗ 13.264 ±0.10
P4 282.40 ±80.38∗∗ 174.60 ±64.55 48.25 ±16.62∗∗ 13.298 ±0.08
KN: normal control; K−: negative control; P1, P2, P3, and P4 were treated with 25, 50, 75, and 100 mg/kg BW crude okra polysaccharide, respectively. Values
are represented as mean ±SD (n�5). ∗∗P<0.05 compared with KN group. ∗∗∗P<0.05 compared with KN, K−, P3, and P4 groups; TNF: tumor necrosis
factor; IFN: interferon; IL: interleukin.
KN K– P1 P2 P3 P4
Spleen index (mg/g)
Figure 2: Eﬀect of crude polysaccharide from okra pods on spleen
index (mg/g). KN: normal control; K−: negative control; P1, P2, P3,
and P4 were treated with 25, 50, 75, and 100 mg/kg BW crude okra
polysaccharide, respectively. Each bar represents mean ±SD
(n�5). ∗∗P<0.05 compared with normal control. ∗∗∗P<0.05
compared with normal and negative control groups.
Splenocyte proliferation (%)
Figure 3: Eﬀect of crude polysaccharide from okra pods on
splenocytes proliferation (%). KN: normal control; K−: negative
control; P1, P2, P3, and P4 were treated with 25, 50, 75, and
100 mg/kg BW crude okra polysaccharide, respectively. Each bar
represents mean ±SD (n�5). ∗∗P<0.05 compared with KN, K−,
P1, and P2 groups.
NK cell activity (%)
Figure 4: Eﬀect of crude polysaccharide from okra pods on NK cell
activity (%). KN: normal control; K−: negative control; P1, P2, P3,
and P4 were treated with 25, 50, 75, and 100 mg/kg BW crude okra
polysaccharide, respectively. Each bar represents mean ±SD
4Advances in Pharmacological Sciences
response and inﬂammation . S. aureus presented by
active macrophage induces activation of -17. T helper-17
produced cytokine such as IL-17, IL-22, and TNF-α.
Among the cytokines, TNF-αis one of the most important
proinﬂammatory cytokines against microbe. e present
result showed that serum level of TNF-αsigniﬁcantly in-
creased in groups with administration of crude okra poly-
saccharide at doses of 25, 50, and 100 mg/kg BW. e
previous study also reported that crude okra polysaccharides
signiﬁcantly increased TNF-αlevel in RAW264.7 macro-
From the previous study, okra polysaccharide-induced
cytokine production from macrophages through activation
of the transcription factor NFlB . e transcription
factor NFlB exhibits a potent activity in modulating gene
transcription involving TNF-α. is present study of TNF-α
demonstrated that administration of crude okra poly-
saccharides at a dose of 75 mg/kg did not signiﬁcantly in-
crease the level of TNF-α.
Contrast with this result, the highest increase of
phagocytic activity was found in the group with at a dose of
75 mg/kg BW. ere is no relation between active macro-
phages that undergo phagocytosis with production of TNF-α
level in this study. is result indicated that TNF-αwas not
dominantly produced by macrophages. Crude poly-
saccharides from okra pods may induce other cells to
produce TNF-α. Zhou et al.  reported that crude extract
of T. wilfordii strongly inhibits TNF-αand IL-1 production.
e overproduction of TNF-αis related with development of
various diseases . us, we found this result beneﬁcial
because TNF-αis suppressed in higher doses. Insigniﬁcant
level of TNF-αin P3 with dose of 75 mg/kg BW could be due
to its optimal doses. Based on this result, optimal doses of
polysaccharides to increase TNF-αwere lower doses (25 and
50 mg/kg BW). Although P4 showed signiﬁcant result, the
level of TNF-αdid not raise as high as P1 and P2 groups.
is study also showed increase of serum level of IFN-c
but the diﬀerence was insigniﬁcant. Meanwhile, serum level
of IL-23 did not show any diﬀerence. Contrast with serum
level of TNF-α, all of the treatment groups had decreased
serum level of IL-17 compared with normal control group
and negative control group. IL-17 is one of the proin-
ﬂammatory cytokines . ese results suggested that
crude polysaccharides from okra pods could modulate
immune function through promoting and inhibiting cy-
tokines level which help in killing of microbes and control
proinﬂammation cytokine level. In this study, crude
polysaccharides from okra pods increased the serum level
of IFN-cand controlled serum level of IL-17 and IL-23
possibly to prevent healthy cells from becoming damaged.
Both IFN-cand IL-17 are proinﬂammatory cytokines, and
overexpression of proinﬂammatory cytokines will induce
Lymphocytes circulated in the blood and the lymph are
also found in large numbers in lymphoid tissues or lymphoid
organs. e spleen is a secondary lymphoid organ, a place for
maintaining mature naive lymphocytes . Spleen weight
and spleen index are changed in response to the nonspeciﬁc
immunity. It has been reported that immunomodulator can
enhance spleen index . is result showed that crude
polysaccharides from okra pods at doses of 50, 75, and
100 mg/kg signiﬁcantly increased the spleen index. is result
demonstrated that polysaccharides from okra pods stimulate
the immune system by inducing proliferation of immune
cells. erefore, we investigated the eﬀect of polysaccharides
from okra pods on the splenocytes proliferation.
Splenocytes are immune cells in the spleen. Splenocytes
consist of T cell, B cell, macrophages, and dendritic cells .
is result showed that splenocytes proliferation was sig-
niﬁcantly increased in groups treated with polysaccharides
from okra pods at doses of 75 and 100 mg/kg. e previous
study also demonstrated an increase of splenocytes pro-
liferation by administration of polysaccharides from Den-
drobium huoshanense . Recent studies indicated that
crude polysaccharides with higher doses increase splenocyte
proliferation. Splenocyte proliferation response is related to
improved T- or B-lymphocyte immunity which could be an
indicator of immune activation . As we report in this
study that crude polysaccharides from okra pods induced
spleen index, increase of spleen index may occur due to
rising of splenocytes proliferation.
As a member of lymphocyte class, NK cells are best
known for their nonspeciﬁc killing of tumor cells, and there
is evidence for their role in controlling infection in the
earliest phases of the body’s immune response . ey can
react against and destroy target cells without the help of
either major histocompatibility complex- (MHC-)
dependent-recognition or prior sensitization, but by exo-
cytosis of perforin-containing granules .
erefore, an NK cell activity assay is a routine method
for analysis of a patient’s cellular immune response in vitro
and can also be used to test the antitumor activities of
possible drugs . In this study, we further investigated NK
cells activity using human hepatocytes cell line. Our result
showed that NK cells activity was able to be restored like
normal control group but the diﬀerence was not statistically
signiﬁcant. Possibly, polysaccharides target macrophages
rather than NK cells.
Immunomodulatory activities of polysaccharides may be
due to direct or indirect interaction with immune system
components. Complement proteins and monocytes, mac-
rophages, dendritic cells, neutrophils, and lymphocytes have
been reported as target responding to polysaccharides
[27–29]. Binding of polysaccharides to speciﬁc recognition
receptors on immune cells trigger diverse signaling pathway
and responses .
e results of the present study all together showed that
crude polysaccharides of the okra pods had the potential to
enhance the immune response of some immune compo-
nents. Treatment groups with higher doses of crude okra
polysaccharides increased phagocytic activity, spleen index,
and splenocytes proliferation. Meanwhile, treatment groups
with lower doses of crude polysaccharides from okra pods
showed the highest signiﬁcant rising of TNF-αlevel. Con-
trast with other results, treatment groups with higher doses
of crude okra polysaccharides showed decrease level of IL-17
as response to prevent overexpression of proinﬂammatory
Advances in Pharmacological Sciences 5
Based on this study, crude okra polysaccharides could act
as an immunomodulator. Crude okra polysaccharides had
both immunostimulator activity and immunosuppression
activity. Most of the immune components examined in this
study showed signiﬁcant increase and decrease at the doses
of 75–100 mg/kg. Crude okra polysaccharide could enhance
immune response, showed with rising of phagocytic activity,
spleen index, splenocytes proliferation, and TNF-αlevel.
Crude okra polysaccharides could suppress immune re-
sponse (immunosuppression), showed with decrease of IL-
We concluded that crude polysaccharides from okra pods
could enhance phagocytic activity, spleen index, splenocyte
proliferation, and TNF-αlevel, but decrease IL-17 level as
a response to prevent overexpression of proinﬂammatory
cytokines. is study suggests that crude polysaccharides
from okra pods grown in Indonesia could act as an eﬀective
compound to improve immune response.
e data used to support the ﬁndings of this study are in-
cluded within the article.
Conflicts of Interest
e authors declare that there are no conﬂicts of interest
regarding the publication of this paper.
is study was ﬁnancially supported by applied research of
pre-eminent college or Penelitian Terapan Unggulan Per-
guruan Tinggi (PTUPT), ﬁscal year 2017, No.
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