The Effectiveness of Immunostimulant from Ethanol Extract of

Parasite Coffee Leaves in Male Rat with Slow Type Hypersensitivity

Method

Rahmad Gurusinga

, Tati Murni Karokaro

, Iskandar Markus Sembiring

, Sri Wulan

, Novita Br

Ginting Munthe

, Romauli Anna Teresia Marbun

and Suci Wulandari

Faculty of Nursing and Physiotherapy, Institut Kesehatan Medistra Lubuk Pakam, Sumatera Utara, Indonesia

Faculty of Midwifery, Institut Kesehatan Medistra Lubuk Pakam, Sumatera Utara, Indonesia

Faculty of Pharmacy, Institut Kesehatan Medistra Lubuk Pakam, Sumatera Utara, Indonesia

Keywords: Ethanol Extract of Parasite Coffee, Volume Reduction of Swelling on a rat's feet, Slow Type

Hypersensitivity.

Abstract: This research aimed to determine the effectiveness of immunostimulant from ethanol extract of parasite

coffee leaves with four dose variants. This research used 18 male white rats with 200 grams of weight

divided into 6 treatment groups. The rats were induced with Escherichia coli for 7 consecutive days intra-

peritoneum, then on the 7th day gave a mark on the rat's feet that had been measured (V0). Ethanol extract

of parasite coffee leaves with 50mg / kg weight, 100mg / kg weight, 200mg / kg weight, 400mg / kg weight,

0.5% CMC Na suspension, and 25 mg / kg weight STIMUNO suspension as positive control were given

orally on 8th day, after 24 hours of administration the volume of the rat's feet was measured again, then all

data from each group was processed by using the ANOVA test. The results showed that the ethanol extract

of the parasite coffee leaves proved to be effective as an immunostimulant. This is attested by reduction in

the volume of swelling on rat's feet which tested by the slow type hypersensitivity. It was said to be

effective because it had an approaching value positive control that is 25 mg stimuno starting from 200 mgkg

/ weight dose to 400 mgkg / weight. With an average reduction in the volume of swelling 1.4 mm for dosage

of 400 mgkg / weight and 1.2 mm for dosage of 200 mgkg / weight.

1 INTRODUCTION

The commonly treatments of the Cancer are surgery,

radiotherapy and chemotherapy, but there are not

getting the optimal results yet from those therapies.

Each of these therapies has several side effects that

tend to harm the patient. The failure that often

occurs in the treatment of this cancer, mainly

through chemotherapy, is caused by the low

selectivity of anticancer drugs against normal cell.

Moreover, the failure of chemotherapy is also

caused by cancer cell resistance to

chemotherapeutic agents. Treatment of this disease

requires an aggressive and innovative approach to

the development of new treatments that require the

role of immunomodulators to enhance the immune

system (Rahim, Suartha and Sudimartini, 2017).

Cancer treatments that are commonly done are

surgery, radiotherapy and chemotherapy, Optimal

results have not been obtained from all of the three

types of therapy. Every therapy has several side

effects that endangers the patient. Failures that often

occur in treatment the cancer, therapy through

chemotherapy is due to low selectivity anticancer

drugs against normal cells (Yulianti et al., 2018).

This resistance phenomenon has consequences for

increasing dosage of therapeutic. It can be called a

Multi-Drug Resistance (MDR) phenomenon that can

increase the level of toxicity of drugs that was used

for therapy. Resistance to therapy is a major problem

in cancer. Even if the treatment is initially

successful, the tumor often remains stubborn.

Chemotherapy treatments are increasingly modern

even though there is no 100% effective cancer

treatment for treating cancer. Drug resistance is

influenced by several factors including individual

factors such as somatic genetic cells in

tumors.Resistance is a common thing in cancer and

as cancer therapy becomes more effective, drug

Gurusinga, R., Karokaro, T., Sembiring, I., Wulan, S., Munthe, N., Marbun, R. and Wulandari, S.

The Effectiveness of Immunostimulant from Ethanol Extract of Parasite Coffee Leaves in Male Rat with Slow Type Hypersensitivity Method.

DOI: 10.5220/0009973504990506

In Proceedings of the International Conference on Health Informatics and Medical Application Technology (ICHIMAT 2019), pages 499-506

ISBN: 978-989-758-460-2

499

resistance is more frequent. The cause of this

resistance is due to the presence of compounds in the

body that detect anti-cancer drugs in the cell to expel

it. Apoptosis, induction and detoxification also play

a role in the formation of body resistance to

drugs.The mechanism in which cancer drug

resistance functions provides important information

in avoiding increasing levels of resistance to

chemotherapy. In addition, resistance also has

implications and pharmacokinetics for drugs that are

generally used (Brown et al., 2016).

Indonesia is rich in biodiversity that has the

potential to be developed as a drug or drug raw

material that the function as an antioxidant with low

cytotoxicity. The research on plants such as herbs,

spices, and plants that ride on other plants, such as

parasite is widely carried out because of its potential

(Wulandari et al., 2019). As an antioxidant. In

Europe, parasites especially the leaves, which have

been studied to be used as drugs in the treatment of

cancer deadly cure. In this parasite leaf there are

various kinds of substances that can help treat this

dangerous disease. Since the 16th century,

Europeans have used parasites as a powerful herbal

remedy for treating neurological disorders such as

epilepsy. For someone who has epilepsy, he tends to

have difficulty controlling his own body and uses

the benefits of parasitic leaves which are believed to

be very effective in overcoming this health problem

(Kumolosasi et al., 2018). Parasite is a semi-

parasitic plant, which was originally considered a

detrimental plant because it damages commercial

plants. But “benalu” has the potential as a medicinal

herb. Traditionally a number of parasitic species

since ancient times have been used to prevent and

treat various diseases including as cough medicines,

cancer, diuretics, anti-inflammatory, antibacterial,

wound or mold infection. The parasite is a type of

plant that propagates on other plants and takes food

from attached plants. Coffee parasite is a type of

plant that lives attached to the coffee plant as its

host. In general, this parasitic plant grows well in

regions that have a tropical climate like in Indonesia.

Many parasites grow on several other types of plants

such as mangoes, coffee, starfruit and many others.

In the past, not many people knew the benefits of

this parasite coffee plant because it was considered

to interfere with the plants that were used as hosts.

Because this process of foraging from parasites

originates from the host that is attached to it, the

plant that is attached becomes lack of food reserves

and cannot grow properly (Ojezele, Erhirhie and

Arojojoye, 2016).

One of the plants that is very good to be a host is

a coffee plant.The parasite of this coffee is usually

attached to the branches and also the trunk of the

coffee tree. Often this plant is found in the

Mandiling Natal area in North Sumatra. The types of

parasites attached to this coffee plant include

parasites which have many benefits that can be used

to cure various diseases. As the attached coffee

plant, parasites also absorb some of the contents of

the coffee plant so the parasite has more or less the

same content in the coffee plant. Usually coffee

plants that have the sap can eventually be used as an

alternative to treating cancer through parasites.

Chemical content that contained in parasites are

phenolic, tannin, amino acid, carbohydrate, alkaloid

and saponin. Phenolic compounds are very active as

antioxidant. Phenolic compounds have a structure

that can easily contribute hydrogen or electrons to

acceptors such as reactive oxygen species or peroxyl

groups from fat, so they can reduce the activity of

oxygen and peroxyl radicals (Ojezele, Erhirhie and

Arojojoye, 2016). Flavonoid is a polyphenol

compound that is found in epidermis of the leaves of

fruit peels and has an important role in human life as

an antioxidant, antimutagenic, antineoplastic and

vasodilator activity. a compound consisting of 15

carbon atoms which are generally spread over the

plant world. More than 2000 plant-derived

flavonoids have been identified, but there are three

groups commonly studied, namely anthocyanin,

flavonols, and flavones (Yulianti et al.,

2018).\Chemical contents that contained in parasites

are flavonoid, tannin, amino acid, carbohydrate,

alkaloid, and saponin. Based on various researches,

compounds in parasites that are thought to have

anticancer activity are flavonoids, which are

quercetin which are inhibitors of DNA enzymes for

cancer cell topoisomerase. Quercetin is the main

flavonoid compound contained in the parasite. The

compound is a taxonomic marker of the

Loranthaceae family (Hueza et al., 2019).

Parasite of coffee is a type of plant whose life

does not require soil for its media. It lives as a

parasite, sticks to the host cell, and absorbs the

nutrients its has so that cause death to the host cell.

The presence of Chlorophyll causes parasites have

the ability to carry out photosynthesis. However, this

plant is not able to take water and nutrients directly

from the soil which makes it as a parasite plant

(Poelman et al., 2019). The using of parasites as

traditional medicine has been known for a long time

to cure various diseases. Parasite is used by the

society as an anti- inflammatory drug, pain reliever

(analgesic), antivirus, anticancer, etc. For example

ICHIMAT 2019 - International Conference on Health Informatics and Medical Application Technology

500

tea’s parasite and mango’s parasite which are used

as the cure for cancer (Choi et al., 2019). The

flavonoid content in coffee’s parasite can be

optimized in the extraction process by defeating

(Yulianti et al., 2018). Definitely, also tumor and

immune cells, including tumor associated

macrophages and tumor-infiltrating lymphocytes (of

both the innate and the adaptive arm of the immune

system) can produce these factor (Poggi, Varesano

and Zocchi, 2018).

The immune system is a body's defense system

that has the function to protect, maintain and destroy

antigens such as bacteria, viruses and other

microorganisms that can cause various diseases. One

effort to maintain the immune system is by giving

(Putra, Azizah and Nopriyanti, 2020). The immune

response is the response of the immune system to

objects or substances that are considered foreign. So

there are two types of immune responses that might

occur, namely the nonspecific immune response and

the specific immune response (Emelda et al., 2018).

Immune system compilation exposed to substances

that are considered foreign, then there are two types

of immune responses that might occur, non-specific

and specific immune response. Non-specific

immune response is immune congenital (innate

immunity) in the sense that the response to foreign

substances can occur in the body has never been

exposed to these substances. Meanwhile, a special

immune response is acquired immune responses that

arise against certain antigens (Xue et al., 2019).

Immunomodulation is a compound that can affect

the humoral and cellular immune systems. There are

two types of immunomodulation, namely

immunostimulator (boosting the immune system)

and immunosupressor (suppressing the immune

system). Some compounds that contained in plants

have immunostimulatory and immunosuppressive

effects. Immunomodulation are related to

macrophage activity and capacity (Yulianti et al.,

2018). Macrophage is one of the cells that play an

important role in the immune response, both

functional roles in phagocytosis and their role as

Antigen Presenting Cells (APC). In carrying out

these two roles, the endogenous mediator aid such as

cytokines is definitely needed. While the need for an

exogenous mediator such as carotene and flavonoid

still needs thorough research (Ofokansi et al., 2018).

A substance that acts as an enhancer or booster

immune can be obtained by using of herbs that have

efficacy as an immunostimulant. One of the herbs

which used is ethanol extract of the parasite leaves

of coffee which according to previous researchers

has the potential as an immunomodulation with high

antioxidant activity (Suparman, Kadarusman and

Situmeang, 2018).

Immunomodulation is a compound that can

affect the humoral and cellular immune systems.

There are two types of immunomodulation, namely

immunostimulator (boosting the immune system)

and immunosupressor (suppressing the immune

system). Some compounds that contained in plants

have immunostimulatory and immunosuppressive

effect, Immunomodulation is related to activity and

capacity of macrophage (Rahim, Suartha and

Sudimartini, 2017). Water extract of ethanol extract

from parasite leaves of coffee

(LoranthusferrugineusRoxb.) Active Test of immune

system activity can be done by various methods,

namely by looking at phagocytosis activity using

carbon clearance methods, slow type

hypersensitivity response, and hemagglutination test

for antibody titer (Zhang et al., 2019). The

effectiveness of the immune system test can be

carried out with the antibody titer method

determined based on the last dilution where the

antibody is still detected through visually involved

hemagglutination. The antibody titer value is

transformed with [2log (titer) +1] (Marbun, Suwarso

and Yuandani, 2018). To obtain the compound of

flavonoids, the parasite leaf powder was extracted

first with methanol, then the dried methanol extract

was extracted again with ethyl acetate and ethyl

acetate extract was partitioned with n-hexane.

Residue that insoluble with n-hexane is a total

flavonoide. Total flavonoids were hydrolyzed with

HCl 2 N to obtain aglycone flavonoids. Methanol

extract, ethyl acetate and n-hexane extract

phytochemical screening to determine the class of

metabolites contained therein, and carried out

antioxidant activity (Eriani et al., 2018). Based on

the above considerations, the authors feel it is

important and necessary to test theimmunostimulator

effect of ethanol extracts of the parasite leaves of

coffee (LoranthusferrugineusRoxb.) in male rats.

Therefore, scientific research such as

immunostimulator research and testing in

pharmacology is needed.

2 METHOD

The tools and materials used rotary evaporators,

mercury water pletismometers, vapor cups, glass

beakers, analytical weights, reaction tubes, CMC

Na, 96% alcohol, Na C l, coffee leaf extract. The

process of taking and preparing samples can be seen

in Figure 1. The animals used 18 white rats about

The Effectiveness of Immunostimulant from Ethanol Extract of Parasite Coffee Leaves in Male Rat with Slow Type Hypersensitivity Method

501

150-200 grams which are treated by unifying food.

Animal examples used for experiments can be seen

in Figure 2a. The dosage used in the research is

based on previous research on the activity and

vasorelaxant of the parasite leaves of 50mgkg

/weight, 100mgkg/weight, 200mgkg /weight, and

400mgkg/weight. Each group was made up of three

rats. The process of giving ethanol extract of

parasitic leaves of coffee (EEBK) to rat can be seen

in Figure 2b.

Figure 1: Taking and preparing sample

(a) (b)

Figure 2: (a) Animal experiment; (b) Giving (EEBK) to rat

Weighed the extract as much as 50mg, was

added to the mortar, then poured a little 0.5% CMC

Na suspension while crushed until homogeneous,

afterhomogeneous it was put into a 100ml

volumetric flask. Phytochemical is used to test

whether or not of secondary metabolites in plants

qualitatively. The method that is used for searching

and finding bioactive compounds is

phytopharmacologic screening approaches. Alkaloid

examination. The sample was weighed as much as

0.5 g then added 1 ml of 2N hydrochloric acid and 9

ml of distilled water, heated over a water bath for 2

minutes, cooled and filtered. The filtrate obtained

was used for the alkaloid test. Then divided into 3

tubes, each tube added 2 drops of reagent mayer, 2

drops of bouchardat reagent, 2 drops of dragendroff

reagent. Alkaloid is positive if there are 2 to 3

deposits in each experiment.

Flavonoid examination: The sample was

weighed 5g, then added 10 ml of hot water, boil for

5 minutes and filtered under heat. Into the filtrate

were powdered magnesium and 1 ml of concentrated

hydrochloric acid and 2 ml of amyl alcohol. Shaken

and allowed to separate. Flavonoid is positive if

there is a yellow, orange or red color in the alcoholic

layer. Tannin examination: The sample was

weighed as much as 1g boil for 3 minutes in natural

distilled water then cooled and filtered. 1-2 drops of

iron (III) chloride 1% w / v are added to the filtrate.

If there is a blackish blue or blackish green indicates

tannin. Saponin experiment: The sample is weighed

0.5g and put into a test tube, then added 10ml of hot

water, cooled, then shaken vigorously for 10

minutes. If a stable 1- 10cm high foam is formed not

less than 10 minutes does not disappear with the

addition of 1 drop of 2N hydrochloric acid then it

shows the presence of saponin. Glycoside

experiment: The sample was weighed 3g and then

mixed with 30ml alcohol-water (7: 3) and 10ml of

2N hydrochloric acid, refluxed for 2 hours, cooled

and filtered. Taken 20 ml of filtrate, 25 ml of

distilled water and 25 ml of lead (II) acetate of 0.4

M are added, shaken, allowed to stand for 5 minutes,

then filtered. The filtrate was filtered with a mixture

of chloroform-isopropanol (3: 2) 3 times. Anhydrate

sodium sulfate is added to the extract, filtered and

evaporated at a temperature of no more than 50ºC.

The remainder is dissolved with 2 ml of methanol,

the remaining solution is put into a test tube,

evaporated on a water bath. To the remaining, added

2 ml of water and 5 drops of molish reagent. Be safe

to added 2 ml of concentrated sulfuric acid, a purple

ring formed at the liquid limit indicates the presence

of a sugar bond. Steroid examination: The sample

was weighed as much as 1g, macerated with 20 ml

of n-hexane for 2 hours, filtered. The filtrate was

evaporated in a vaporizer cup and the remaining

concentrated sulfuric acid reagent was added

through the wall of the cup. If a purple or red color

turns into purple or blue, green, it indicates the

presence of steroids / tri terpenoids.

The phytochemical test results of the parasite

leaves sample contained alkaloids, tannins, saponins,

glycosides and flavonoid. Phytochemical test results

on leaves are slightly different from those ever

reported by muamar muulian who stated that

secondary metabolites contained in methanol

extracts of the parasite leaves of coffee are alkaloids,

flavonoids, terpenoids, and tannins. This is expected

because the samples obtained are from different

places so that the content of secondary metabolite is

different.(Yulianti et al., 2018). Phytochemical

screening test results can be seen in Table 2 and

recording the results of the swollen volume of rat

feet can be seen in Table 1.

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Table 1: Recording of the results of the volume of

swelling of the rats feet.

Table 2: Phytochemical screening test

Test animals used 18 male rats with a weight of

150-200 grams. Before treatment, the experimental

animals were first conditioned for 2 weeks in a good

cage to adjust their environment and uniform food.

The animals were grouped into 6 groups including 1

group with 0.5% CMC Na absorption, the second

group with a 25mg stimuno suspension, the third

with 50mgkg /weight ethanol extract suspensions,

the fourth with a leaf ethanol extract suspension

parasite’s coffee with a dose of 100mgkg/weight, the

fifth with ethanol extract suspension of the parasite

leaves of coffee at a dose of 200mg/weight, the sixth

with the suspension of ethanol extract of the parasite

leaves with a dose of 400mg/weight. Each group of

experimental animals were injected e.coli inoculums

intravenously as antigens with 0.1 inoculum,

treatment was started on day 0. The process of the

slow type hypersensitivity testing method can be

seen in Figure 3.

Figure 3: Slow Type Hypersensitivity Testing Method.

Homogeneity and normality are determined by the

research results to determine the statistical analysis

used. If there are differences, proceed by using the

Post Hoc Tuckey test to find out which variables

have differences. Data were analyse using the one-

way ANOVA test to determine the average

difference between treatments. Based on the

significance value of p ≤ 0.05, it is considered

significant. The type of research used is Laboratory

Experiment. This research uses Anova One Way

parametric test. The sample which used in this study

was the Ethanol Extract of Parasite coffee Leaves

(LoranthusferrugineusRoxb). The data obtained

were analyzed using SPSS with the Anova test.

Continued by the Post Hoc Tukey test to see

variables that have a significance value of p <0.05.

3 RESULT AND DISCUSSION

Phytochemical test results of the parasite leaves of

coffee containing alkaloids, tannins, saponins,

glycosides and flavonoids. Phytochemical test

results on leaves are slightly different from those

ever reported by Yullian Muamar who stated that

secondary metabolites contained in methanol

extracts of the parasite leaves of coffee are alkaloids,

flavonoids, terpenoids and tannins. This is expected

because the samples that obtained are from different

places so that the content of secondary metabolite is

different.(Yulian, 2018). From 1500 grams of dried

samples macerated using a rotary evaporator to get

30 grams of ethanol extract of parasitic coffee. Thick

extract has a distinctive odor, black in color.The

result of this research of ethanol extract of parasite

coffee leaves is given for 7 days at a dose of 50 mg /

Before After Deviation

1 2,12 1,2 0,92

2 2,12 1,2 0,92

3 2,13 1,2 0,93

1 2,12 1,2 0,92

2 2,12 1,14 0,98

3 2,13 1,19 0,94

1 2,12 0,92 1,2

2 2,12 1,02 1,1

3 2,13 0,93 1,2

1 2,12 0,82 1,3

2 2,12 0,72 1,4

3 2,13 0,83 1,3

1 2,12 0,45 1,67

2 2,12 0,47 1,65

3 2,13 0,48 1,65

1 2,12 1,47 1,65

2 2,12 1,4 0,63

3 2,13 1,48 0,65

BLANKO

0.95

1,2

1,4

1,6

0,64

50mg/kgweight

100mg/kgweight

200mg/kgweight

400mg/kgweight

STIMUNO

No Doses Rat Average

Swellingvolumeofratfeet

0,92

The Effectiveness of Immunostimulant from Ethanol Extract of Parasite Coffee Leaves in Male Rat with Slow Type Hypersensitivity Method

503

BW and a dose of 100 mg / BW still provide effects

such as negative control. However, at a dose of 100

mg / BW has begun to lead to positive control, at a

dose of 400 mg / BW the measurement results of the

swelling feet volume of rat approach the positive

control of stimuno with an average value of 1.6, with

an average of 1.4. We strongly encourage authors to

use this document for the preparation of the camera-

ready. Please follow the instructions closely in order

to make the volume look as uniform as possible

(Breda et al., 2019). Based on the results of the

research that has been done by varying the dosage of

ethanol extract of parasites of coffee leaves by

measuring the volume of rat swelling using a

mercury pletismometer, using the slow type

hypersensitivity method is found in Figure 4.

Figure 4: Swelling volume of rat feet.

The results of macroscopic examination carried out

on the parasite leaves of coffee

(LoranthusferrugineusRoxb.) Namely the top surface

is smooth and green, the lower surface is weak

green. The macroscopic results of the form of coffee

parasites are slim-shaped roots, radiating at the host

and dull in color. Long stems of plants upright dull

green small, oval shaped leaves that have a dark

green color with a slightly rough surface. There are

small seeds - interrupted between leaf stalks and

stems, small seeds shaped like the contents of a

pencil, have a short tentacle, macroscopic and

microscopic examination carried out on parasites of

coffee leaves in Figure 5 and Figure 6.

Figure 5: Macroscopic examination carried out on

parasites of coffee leaves.

Description Figure 5: Leaf stalk parenchyma (1),

Leaf parenchyma with calcium oxalate crystal (2),

Hair cover (3), Palisade fragments and leaf

parenchyma (4), Flower petals fragments with petals

(5), Pericarp parenchymal fragments (6), Trachea

(7), Pollen (8).

Figure 6: Microscopic examination of the parasites of

coffee.

Description Figure 6: Hair cover (1), Kolenkim (2),

Upper epidermis (3), Palisade parenchyma (4),

Spongy tissue (5), Floem (6), Xilem (7), Lower

epidermis (8).

The results of the research were obtained from the

administration of ethanol extract of the parasite

leaves of coffee (LoranthusferrugineusRoxb) at a

dose of 50 mgkg /weight, 100 mgkg / weight, 200

mgkg /weight, and 400 mg/weight for 7 days

showed a decrease in swelling volume, this can be

seen in the Table 3. However, the results of several

dose variants showed an average difference

between each treatment.Comparison of the

average value of eachtreatment that is at a dose of

50 mg / BW and 100 mg / BW that tends to have

an average value approaching negative control,

namely CMC Na, but at a dose of200 mg / BW to a

dose of 400 mg / BW the average value changes

towards the direction of positive control. There is a

difference in the average value of each test group

that has been done. Then it will be continued by

using the post hock tukey test to find out which

variable has a difference based on the significance

value<0.005 which is considered significant, this can

be seen in the Table 4.

Table 3: Changes in swelling feet volume of rat

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Table 4: Results of data analysis.

From the results of the post hock tukey test

between the negative control group, the 50 mgkg /

weight dose and the 100 mgkg /weight dose had an

effect but was not significant compared to the 200

mgkg /weight dose and the 400 mg kg / weight dose

which gave an average value close to the 25 mg

Stimuno positive control. Decrease in the volume of

swelling of rat's feet is suspected because in the

extract contained secondary metabolites such as

alkaloids, flavonoids, tannins, saponins, and

glycosides contained in coffee parasites leaves

(LoranthusferrugineusRoxb). This compound

belongs to the group of polyphenols which besides

having biological functions such as improving

glucose metabolism as well as antioxidants and

immunostimulants.

4 CONCLUSIONS

The results showed that the using of ethanol extract

of parasite leaves at a dose of 400 mgkg/weight

could reduce the swelling volume of rat's feet and

was effective as a significant immunostimulant can

be seen from the significance value of р <0.005. The

recommended dose used as immunostimulant is the

highest dose. At the highest dose in this research, it

has the same effectiveness with positive controls that

already exist in the market, namely

immunstimulants.

ACKNOWLEDGEMENTS

This Research is Supported by Institut Kesehatan

Medistra Lubuk Pakam, Medistra Foundation,

Institut Kesehatan Deli Husada, and Sembiring

General Hospital Foundation, Indonesia. My deepest

thanks to Institut Kesehatan Medistra Lubuk Pakam

and Institut Kesehatan Deli Husada for providing

the opportunity for all lecturers on this beloved

campus to gain Knowledge in Writing the Scientific

Journal and provides new Experiences in holding the

International Conferences. Hopefully with the

implementation of this activity in the future I can

contribute to the progress of Institut Kesehatan

Medistra Lubuk Pakam and Institut Kesehatan Deli

Husada.

REFERENCES

Breda, S. V. Van et al. (2019) ‘Computational

Methodologies for the in vitro and in situ

Quantification of Neutrophil Extracellular Traps’,

Frontiers in Immunology, 10(July), pp. 1–7. doi:

10.3389/fimmu.2019.01562.

Brown, M. et al. (2016) ‘Seasonal immunoregulation in a

naturally- occurring vertebrate’, BMC Genomics.

BMC Genomics, 17(369), pp. 1–18. doi:

10.1186/s12864-016-2701-7.

Choi, M. et al. (2019) ‘Anti-inflammatory effects of a

methanol extract of Dictamnus dasycarpus Turcz . root

bark on imiquimod-induced psoriasis’, BMC

Complementary and Alternative Medicine. BMC

Complementary and Alternative Medicine, 19(347),

pp. 1–8.

Emelda, A. et al. (2018) ‘Immunomodulatory Effect of

Ethyl Acetate Extract of Permot ( Passiflora Foetida L

.) Leaf against the Secretion of Antibody and Delayed

Type Hypersensitivity in Vivo Journal of Global

Pharma Technology Immunomodulatory Effect of

Ethyl Acetate Extract of P’, Journal of Global Pharma

Technology, 1(January 2019). doi:

10.31219/osf.io/yq6vh.

Eriani, K. et al. (2018) ‘IMMUNOSTIMULATORY

EFFECT OF METHANOL EXTRACT OF

FLAMBOYANT LEAF (Delonix regia (Boj.ex Hook)

Raf) IN MICE’, Jurnal Natural, 18(1), pp. 44–48. doi:

10.24815/jn.v18i1.9830.

Hueza, I. M. et al. (2019) ‘Immunomodulatory effect of

Cynara scolymus ( artichoke ) in rats’, Phytotherapy

Research, 33(September 2018), pp. 167–173. doi:

10.1002/ptr.6210.

Kumolosasi, E. et al. (2018) ‘Immunostimulant activity of

standardised extracts of Mangifera indica leaf and

Curcuma domestica rhizome in mice’, Tropical

Journal of Pharmaceutical Research, 17(1), pp. 77–84.

Marbun, R., Suwarso, E. D. Y. and Yuandani, Y. (2018)

‘IMMUNOMODULATORY EFFECTS OF

ETHANOL EXTRACT OF ARTEMISIA

VULGARIS L . IN MALE RATS’, Asian Journal of

Pharmaceutical and Clinical Research, 11(1), pp. 245–

247.

Ofokansi, M. N. et al. (2018) ‘Immunomodulatory Effects

of Phyllanthus muellerianus : A Mechanistic Journal

of Clinical & Cellular Immunomodulatory Effects of

Phyllanthus muellerianus : A Mechanistic Approach’,

Journal of Clinical & Cellular Immunology, 9(5), pp.

1–8. doi: 10.4172/2155-9899.1000565.

Ojezele, M. O., Erhirhie, E. O. and Arojojoye, O. A.

(2016) ‘Effects of Viscum album ( mistletoe ) from

three host plants ( cocoa , kola and coffee ) on semen

The Effectiveness of Immunostimulant from Ethanol Extract of Parasite Coffee Leaves in Male Rat with Slow Type Hypersensitivity Method

505

quality of wistar albino rats’, International Scientific

Organization, 2(1), pp. 109–113.

Poelman, E. et al. (2019) ‘Effects of immunomodulation

in classic infantile Pompe patients with high antibody

titers’, Orphanet Journal of Rare Diseases. Orphanet

Journal of Rare Diseases, 14(71), pp. 1–11.

Poggi, A., Varesano, S. and Zocchi, M. R. (2018) ‘How to

Hit Mesenchymal Stromal Cells and Make the Tumor

Microenvironment immunostimulant Rather Than

immunosuppressive’, Frontiers in Immunology,

9(February), pp. 1–17. doi:

10.3389/fimmu.2018.00262.

Putra, B., Azizah, R. N. and Nopriyanti, E. M. (2020)

‘Efek Imunomodulator Ekstrak Etanol Herba Krokot (

Portulaca olerace a L .) terhadap Tikus Putih ( Rattus

norvegicus ) Jantan dengan Parameter Delayed Type

Hypersensitivity ( DTH )’, Jurnal Farmasi Galenika

(Galenika Journal of Pharmacy), 6(1), pp. 20–25. doi:

10.22487/j24428744.2020.v6.i1.14106.

Rahim, M. A., Suartha, I. N. and Sudimartini, L. M.

(2017) ‘Efek Imunostimulator Ekstrak Daun Kasturi (

Mangifera Casturi ) Pada Mencit’, Indonesia Medicus

Veterinus, 6(1), pp. 10–19. doi:

10.19087/imv.2017.6.1.10.

Suparman, A. R., Kadarusman, M. and Situmeang, B.

(2018) ‘SENYAWA TRITERPENOID DARI

TUMBUHAN PIRDOT ( Sauralia’, Jurnal ITEKIMA,

3(1), pp. 12–19.

Wulandari, S. et al. (2019) ‘EFEKTIFITAS

IMUNOSTIMULAN DARI EKSTRAK ETANOL

DAUN SIRSAK ( Annona mucirata ) PADA TIKUS

JANTAN DENGAN METODE

HYPERSENSITIVITAS TIPE LAMBAT’, Jurnal

farmasimed(JFM), 2(1), pp. 21–30.

Xue, X. et al. (2019) ‘Dietary Immunostimulant CpG

Modulates MicroRNA Biomarkers Associated with

Immune Responses in Atlantic Salmon (Salmo salar)’,

Journal Cells, 8(1592), pp. 1–22.

Yulianti, R. et al. (2018) ‘The Activity Test of Ethanol

Extract Tea Parasite Herb ( Scurrulla Artopurpurea )

as an Immunostimulator on Wistar Strain Rat

Sensititized with Sheep Red Blood Cell The Activity

Test of Ethanol Extract Tea Parasite Herb ( Scurrulla

Artopurpurea ) as an Im’, Journal of Physics, pp. 1–7.

Zhang, W. et al. (2019) ‘Immunomodulatory E ff ects of

the Meretrix Meretrix Oligopeptide (QLNWD) on

Immune-Deficient Mice’, Journal Molecules,

24(4452), pp. 1–12.

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