rd international conference on advance molecular...

2 3rd International Conference on Advance Molecular Bioscience and

Biomedical Engineering (ICAMBBE) 2016

Malang

http://icambbe.ub.ac.id/1st-international-conference-on-advance-molecular-bioscience-biomedical-engineering-2014-icambbe-2014-september-11-13-2014-malang-indonesia/



3

PL-1.1

Maternal High-Fat Diet Promotes Onset of Diabetes in Rat Offspring

Takeshi OHTA1,3, Yasufumi TORINIWA1, Naruhiko RYUMON2, Nobuhiro INABA2,

Tadaaki HIRAO2, Saori YAMANAKA2, Takayuki MAENO2, Wakako SAKAKIBARA2,

Morio SUMIKAWA2, Kaoru CHIBA2, Akiko NAKAMURA1, Katsuhiro MIYAJIMA1,

Fatchiyah FATCHIYAH3 and Takahisa YAMADA4

1 Japan Tobacco Inc., Central Pharmaceutical Research Institute, Takatsuki, 2 JT Creative Cervice Co., Ltd., Takatsuki office, Takatsuki, Osaka, Japan,

3 Department of Biology, Faculty of Mathematics and Natural Sciences, Brawijaya

University, Malang, East Java, Indonesia, 4Laboratory of Animal Genetics Graduate School of Science and Technology, Niigata

University, Niigata, Japan

The onset and progression of type II diabetes is closely related to

environmental factors, in particular dietary habit. Moreover, the

environmental exposures very early in life can influence the risk for

development of type II diabetes later in life. In this study, we investigated

pathophysiological changes in the pups of maternal Spontaneously Diabetic

Torii (SDT) rats that were fed a high-fat diet (HFD) throughout gestation

and lactation. Maternal SDT rats were continued on HFD for 5 weeks, from

day 8 of gestation to day 21 after birth, and biological analyses of the pups

were performed from 2 to 22 weeks of age. Results of serum lipid levels in

pups from dams fed HFD were higher than pups from dams fed a standard

diet, and the onset of diabetes was significantly accelerated in pups from

dams fed HFD. In pathological analyses, pups from dams fed HFD showed

increases in liver weight and vacuolation of hepatic cells at 2 weeks of age.

In conclusion, the metabolic disorder of lipids and glucose in SDT rats is

closely related to the nutritional condition of dams during the periods of

gestation and lactation

Keywords: diabetic model, high-fat diet, SDT rat.



Malang

PL-1.2

SDT-fatty Rats Show Non-Alcoholic Steatohepatitis (NASH)-Like

Hepatic Lesions

Tomohiko Sasase, Ph.D.

Japan Tobacco Inc, Central Pharmaceutical Research Institute, Japan

To investigate the histological features of the liver in spontaneously

diabetic Torii (SDT) fatty rats compared with age-matched Sprague-

Dawley (SD) rats. Female SDT Leprfa (SDT fatty) rats and age-matched SD

rats were fed ad libitum. Body weight and biochemical parameters, such as

serum glucose, triglyceride (TG), total cholesterol (TC), alanine

aminotransferase (ALT) and aspartate aminotransferase (AST) levels as

well as fatty acid and TG accumulation in the liver were evaluated at 8 wk

of age in the non-fasting state and at 8-wk intervals from 8 to 40 wk of age.

Histopathological examinations of the liver were performed using

hematoxylin & eosin and Sirius Red staining as well as double staining for

ED-1 and toluidine blue (TB). The expression of genes involved in TG

synthesis, inflammation, and fibrosis was examined in the liver. SDT fatty

rats showed significantly increased body weight compared with SD rats.

Serum glucose, TG, and TC levels were significantly higher in SDT fatty

rats compared with SD rats. The serum AST and ALT levels in SDT fatty

rats were significantly elevated at 8 wk of age compared with the levels in

SD rats. Hepatic TG content was marked in SDT fatty rats from 8 to 32 wk

of age. Histopathologically, severe hepatosteatosis accompanied by

inflammation was observed at 8 wk of age, and fibrosis started to occur at

32 wk of age. Furthermore, Sirius Red and ED-1 staining were increased in

the liver at 32 wk of age. Hepatic gene expression related to TG synthesis,

inflammation and fibrosis tended to increase in SDT fatty rats compared

with SD rats, and the gene expression related to TG secretion was

decreased in SDT fatty rats compared with SD rats. Female SDT fatty rats

have the potential to become an important animal model of non-alcoholic

steatohepatitis with type 2 diabetes and obesity.

Keywords:




5

PL-1.3

Creating New Economic Opportunities Trough Reproduction

Technology in Tetra-Helix Perspective: The Case of Artificial

Insemination in East Java Province

drh. Enniek Herwijanti, MP

Ministry of Agriculture/ BBIB Singosari

Beef cattle population in East Java Province that reached 4.12 million head

is the highest beef cattle population in Indonesia and contributed for 32%

of national beef cattle population. That achievement was not reached

briefly. The basic program known as “Intan Sejati” (Artificial Insemination

of one million cows) has been made since the year 2004-2009. The

program was made based on the study and cooperating of East Java

Government- BBIB Singosari and Universities in East Java (the Brawijaya

and Airlangga University) within the triple helix scheme.

Calves which is result of cross-breeding through artificial insemination

immediately demanded by livestock owners who are mostly small-scale

farmers who puts livestock as saving or insurance. Beside of that, the

increasing demand for artificial insemination is immediately followed by

the need for the number of inseminator. Artificial insemination

development in east java has opened up new economic opportunities for the

people in East Java. The high price of calves’ result of artificial

insemination has increased the economic value of livestock, but more

significant is the creation of new jobs as “the inseminators” which until

now has reached 1343 people in all districts/cities in East Java.

Increases need for artificial insemination turned out to have created a new

community, a community that is formed from livestock owners and

inseminator which the majorities are private inseminator as the services

user. The entry of this community into the triple helix scheme causes the

changes of the scheme towards tetra helix or quadruple scheme. Within this

tetra-helics scheme, the initiation still comes from the traditional three

helices (Academic-Government-Bussines), but the users as the 4th helix had

a role not only as consumers but also had a role in providing feedback

information on services also sustainable development of products and



Malang

service also sustainability and improvement of their economic

opportunities.

In its development, the synergies that are formed in the form of tetra-helix

has developed not only traditionally in increasing livestock population and

community economic opportunity, but also evolve toward local germplasm

conservation programs that require increased further research cooperation

in the scheme of tetra-helix synergism.

Keywords:




7

PL-2.1

Vaccine Development – From Discovery to Commercialization

Drs. Adriansjah Azhari, Apt., MM

Research and Product Development

PT. Bio Farma (Persero)

Vaccine is the one of the most effective tools in reducing mortality and

morbidity due to infectious disease. Human beings have benefited from

vaccines for more than two centuries. Immunization is a cost-effective and

life-saving measure in preventing suffering through sickness, disability and

deaths. It has eradicated small-pox, lowered the incidence of polio and

dramatically reduces suffering and deaths from diphtheria, tetanus,

pertussis and measles.

The challenges of vaccine development are not limited to identification of

suitable antigens, adjuvants, formulation and delivery methods, but include

regulatory, technical and manufacturing obstacles that need to be addressed

in translating a vaccine candidate from bench-sale to industrial scale

production and ultimately be available in the clinics.

Decision for the manufacture of new vaccines and technologies must be

based on country disease burden with priorities and on what is affordable

and sustainable. From an industrial perspective, underlying factors such as

choice of technology, availability of knowledgeable human resources,

funding, and delivery system and above all, marketing feasibility, need to

be carefully considered before embarking on the manufacture of new

vaccine.

Vaccine research is long, complex and costly. Because a vaccine is a

biological product made from living microorganism, its development cycle

is quite different from that of a pharmaceutical product involving the

following steps:

1. Identification of disease burden

2. Identification of target antigen

3. Vaccine candidate design compising of, among others, feasibility,

production process, formulation, presentation, product stability.

4. Animal toxicity studies (pre-clinical studies)

5. Human clinical trials

6. Product licensure



Malang

7. Post marketing surveillance

The collaboration among academics, research institution and ultimately

vaccine manufactures, will undoubtedly stimulate and accelerate the

development of new vaccines. The long period of vaccine development

which normally takes 20 years and even longer, may be potentially

reduced.

Keywords:




9

PL-2.2

Erythropoietin as an Essential Hormone for Red Blood Cell

Production

Dr. Adi Santoso

Research Centre for Biotechnology, Indonesian Institute of Sciences, Jln. Raya Bogor Km

46. Cibinong. Bogor

Erythropoietin (EPO) is a glycoprotein hormone that regulates the

formation of red blood cells, and is produced primarily by the kidney in the

adult and by the liver during fetal life. EPO is approximately 30-37 kDa

glycoprotein (exact molecular weight depends on degree of glycosylation)

in which sixty percent of the molecule is an invariant 165 amino acid single

polypeptide chain containing two disulphide bonds. The remaining 40% of

the EPO mass consists of carbohydrate covalently attached at three N-

linked and one O-linked sugar chain at Ser126.

Recombinant human erythropoietin (EPO) is produced as a therapeutic

agent for the treatment of anemia associated with severe kidney damage.

The use of recombinant human EPO is now widely applied for the

treatment of anaemia associated with renal failure, cancer, prematurity,

chronic inflammatory disease and human immunodeficiency virus

infection. The expiration of patents and other intellectual property rights for

originator biologicals over the next decade opens up opportunities for

biosimilars to enter the market and increase industry competition.

Previously, a novel erythropoiesis stimulating protein (NESP) called

darbapoetin-α (DARB) which possesses two additional Nlinked

glycosylation had been studied. It was observed that NESP showed better

in vivo stability and biological activity compared to the unmodified form

(containing only 3N-linked carbohydrate chains). Based on the above

study, we attempted to synthesize recombinant human EPO (rHuEPO) by

generating CHO-K1 and CHO-DG44 cell lines expressing codon-optimized

human epo.

Keywords:



Malang

PL-2.3

Role of functional feeds and feed additives in sustainable animal

production

Prof. Dr. Abdul Razak Alimon

Departement of Animal Science, University Putra Malaysia, Putrajaya, Malaysia

Visiting Professor, Universitas Gadjah Mada Yogjakarta, Indonesia

Functional feed additives are substances that are added to the feed and

serve specific functions that can alter digestion, absorption and metabolism

in the animal, subsequently increase feed efficiency, productivity and

growth. The fluctuation in feed prices, due to climatic feed, marketing and

increased demand and the ban in the use of antibiotics has prompted the

livestock industry to resort to the use of these functional feed additives.

These functional feeds additives include probiotics, prebiotics, enzymes,

herbs and herbals, ionophores, organic acids and other forms such as

vaccines, lecithin and amino acids. These feed additives are usually

included in complete feeds and acts in the way of improving digestion

through improved GIT microbial environment, improved absorption of

nutrients, provide balanced nutrients utilization and assimilation, enhanced

tissue growth and development. Herbs are especially important as they have

been used for centuries in both man and animals. The increasing consumer

demands for organically grown foods and functional animal products, such

as high omega 3 eggs, high antioxidant meat, and probiotics fortified milk

has encouraged the use of herbs and other feed additives in the animal

feeding. This paper discusses the various additives and their role in animal

agriculture in the face of challenges such as global warming and risks

towards human wellbeing.

Keywords:




11

PL-3.1

Transcriptional Regulation of Metabolic Pathways in Medicinal Plants

Ling Yuan, Ph.D., Harold R. Burton

Endowed Professor in Plant Biochemistry, Department of Plant and Soil Sciences,

University of Kentucky, USA

More than half of the 150 most-prescribed medicines have at least one

compound derived from plants. Catharanthus roseus (Madagascar

periwinkle) is the major source of a number pharmaceutically important

terpenoid indole alkaloids (TIAs), including the anticancer drugs,

Vincristine and Vinblastine. However, the molecular mechanism

underlying the regulation of TIA biosynthesis is not well understood. The

hormone, jasmonic acid (JA), is a major elicitor of TIA biosynthesis. I will

provide insights into transcriptional regulation of phytohormone signal

cascades that control metabolic pathways of medicinal plants.

Keywords:



Malang

PL-3.2

A systematic analysis of high temperature response in Arthrospira

platensis

Kalyanee Paithoonrangsarid, Ph.D

Biochemical Engineering and Pilot Plan Research and Development Unit, King Mongkut’s

University of Technology Thonburi, Bangkok, Thailand

Arthrospira is a cyanobacterium that has been commercially cultivated in a

large scale and used in many applications such as healthy food products,

animal feed supplements and nutraceuticals due to its high-nutritional value

including rich of protein content, gamma-linolenic acid, vitamins and

minerals and other bioproducts. In outdoor environment, multiple stresses

occur simultaneously, leading to the reduction of cell growth and alteration

of biochemical compounds. High temperature and nutrient limitation affect

biomass production of Arthrospira. To advance our understanding of

cellular stress response, genome-wide gene expression in A. platensis C1

under high temperature (42°C) as well as combination of high temperature

and nitrogen depletion was performed by microarray. The results indicated

that cells responded to high temperature stress by changing the level of

transcripts of several genes involved in different metabolisms. The

transcript levels of several genes encoding heat shock proteins and genes

involved in signaling transduction and carbon metabolism were transiently

induced by high temperature stress, whereas the transcript levels of genes

encoding nitrate and nitrite transporters were repressed. Furthermore, the

combined stress of high temperature and nitrogen depletion had strong

effects on the transcript levels of several genes in the cells. The results

showed that transcript levels of genes involved in phycobilisome

degradation and transcriptional regulation were rapidly up-regulated, while

the transcript levels of genes associated with photosynthesis decreased.

Moreover, the co-expression network based on transcriptional profiles was

constructed to gain insight into the regulatory mechanisms for heat-

responsive gene expression. The results provide a better understanding of

high temperature response mechanism and may also pave the way for

further research to improve Arthrospira biomass production under high

temperature

Keywords:






Malang

PL-3.3

Development of Brucellosis detection Kit based on OMP2-Gene to

Prevent Zoonosis in Indonesia

Aulanni’am Aulanni’am1,2,3*, Dyah Kinasih Wuragil1,3, Masdiana C. Padaga1,3, Agung

Pramana W.Marhendra3,4, Wiwiek Tyasningsih5 and Hapsari Mahatmi6

1Faculty of Veterinary Medicines, Brawijaya University, INDONESIA

2Biochemistry Laboratory, Faculty of Sciences, Brawijaya University, INDONESIA 3Reasearch Group of Biopeptide, Immunodetection and Vaccine (BIODET-VAKSIN),

Brawijaya University, INDONESIA 4Department of Biology, Faculty of Sciences, Brawijaya University, INDONESIA

5Faculty of Veterinary Medicine, Airlangga University, INDONESIA 6 Faculty of Veterinary Medicine, Udayana University, INDONESIA

*Corresponding E-mail:

Brucellosis is an important zoonotic disease which affect human’s health. It

also causing significant economic losses worldwide. In Indonesia,

Brucellosis placed third rank of government’s concerning emerging

diseases. Early detection of this disease rely on Rose Bengal Test (RBT),

Complement Fixation Test (CFT) and several ELISA-based technique,

however those tests have several disadvantages. This research aimed to

isolate and characterize OMP2 gene of local Indonesian isolate of Brucella

sp. The bacteria was isolated using standard microbiological methods and

characterized based on biochemical and serological tests. DNA was

isolated and OMP2 gene was further characterized, resulting DNA

fragment of 224 bp. The OMP2 gene was then confirmed by sequencing

and the results were analyzed for homology using BLAST. Result shows

that the OMP2 gene was successfully isolated with high levels (97% ) of

similarity with Brucella abortus in Gene Data Bank

(http://ncbi.nlm.nih.gov). This finding showed sensitivity that may propose

a potential advantages over existing commercial detection kit.

Keywords: Brucellosis, OMP2 Gene, detection kit.




http://ncbi.nlm.nih.gov/

15

IS-1

Flocculation of Chlorella Vulgaris Induced by Chitosan: Role of

Recultivation and Practical Implications

Anik M. Hariati1*, A. Yuniarti1, M. Fakhri1, Nasrullah B. Arifin1, Mega Y. 1 and D.G.R.

Wiadnya1

1Research Group of Aquatic Biofloc, Faculty of Fisheries and Marine Sciences, University

of Brawijaya


Production of microalgae has been very dependent on the application at a

high cost. A potential method to achieve high yield of microalgae was

conducted by separation between microalgae cells and those culture media.

Flocculation is seen as a cheap separation method to obtain high yields. But

some methods of flocculation caused microalgae could not be re-cultivated.

This study was aiming to evaluate the potential of chitosan as a flocculant

for microalgae. The density of microalgae, biomass, chlorophyll

concentration and re-cultivation efficiency on Chlorella vulgaris were

studied using five different doses of chitosan. The results showed that the

chitosan flocculation at 250 ppm gave the best response. The efficiency re-

cultivation reached 80 % after a week storage.

Keywords:



Malang

IS-2

Monitoring the Composition of Bacterial Community by Clone

Analysis during Enrichment Culture

Irfan Mustafa1, Hiroto Ohta2, Takuro Niidome2 and Shigeru Morimura2

1Biology Department, Faculty of Sciences, Brawijaya University 2Graduate School of Science and Technology, Kumamoto University

*Corresponding E-mail: [email protected]

Continuous culture is a useful technique for isolating autotrophic sulfur

bacteria from the environment. However, little is known about the

composition of sulfur bacterial community during growth in continuous

culture before isolation on agar medium. The objective of this study was to

examine the bacterial composition using the clone analysis by targeting 16S

rRNA at several sequential time during growth in continuous culture. A

sampled sediment from Midorikawa tidal flat, Japan was inoculated to

Thiosulfate Mineral Medium in a continuous stirred tank reactor (CSTR).

The medium was prepared to support growth of autotrophic sulfur bacteria.

During the incubation period, the culture was sampled at week 2, 4, and 26

for observing composition of bacterial community in regard of their 16S

rRNA sequences. The observation was conducted by using the clone

analysis. At all sequential sampling time, the culture was dominated by

autotrophic strains (Gammaproteobacteria) representing 80% of the

detected total clones. The remaining clones were identified as heterotrophs.

Prolonged incubation period after two weeks did not significantly affect the

community composition. Heterotrophic sulfur bacteria were not eliminated

in the medium by an extended incubation period.

Keywords: Sulfur bacteria, continuous culture, clone analysis




17

IS-3

The Realization of Gamet Frozen Cells Bank Of Native and Local

Animal Genetic Resources of Indonesia

G. Ciptadi1, S. Rahayu2, Fatchiyah2, Maidaswar3, Sarastina3, Nurhayati3, S. Wahjuningsih1,

E. Herwiyanti3 and MN. Ihsan1

1 Faculty of Animal Husbandry, UB, Indonesia, 2. Faculty of MIPA-UB, and 3. Center of

Artificial Insemination (BBIB) , Malang Indonesia

* Corresponding E-mail:

These study aims were to realize the gamete cell bank of spermatozoa and

oocyte of some specific breeds, superior and potential for conservation of

animal genetic resources of Indonesia. Utilization of frozen cell stock products

can be used for the purposes of providing genetic material for both research

and commercial applications Artificial Insemination (AI). This frozen gamete

cells have the potential to trade at a national, regional and international levels.

These activity as a feasibility study of the conservation of genetic material

from a technical, economic and scientific as well as the growing trade in

anticipation of the possibility of cell-based international level local resources.

The study focused on the nation's local goats and PE. and Senduro main

producing frozen sperm of some local goat breeds that have been selected.

Production is gamete cell in good quality from the point of viability and

fertility post-thawing, genetic and can be used as production materials for the

production process through the application of commercially AI. Genetic

analysis of goats was performed in the district of Senduro, Lumajang East

Java has performed through chromosome and DNA analysis. Meanwhile,

research also was conducted to test cell freezing and post-thawing viability

stock storage in liquid nitrogen. Test fertility studies have been conducted in

vitro testing of sperm quality, while the AI success in field trials was done in

controlled research station. Genetic analysis has been completed and indicates

that the goat Senduro is categorised have normal chromosome.

Implementation of the Ai test has been conducted at research stations and UPT

Singosari, Malang. AI implementation and production of frozen semen goat

both Senduro and PE showed good results according to the standard test of the

quality of spermatozoa, and the implementation result of the AI.

Keywords: AI, Conservation, genetic resources, SNI, frozen sperm, Goat,

Senduro and PE.



Malang

BMB-1

Detection of Hepatitis C Virus RNA in Blood Donors by Nested PCR

Technique

Supiana Dian Nurtjahyani1*, Mochammad Amin2 and Retno Handajani2

1 Faculty of Teacher and Educational Science, Universitas PGRI Ronggolawe Tuban 2 Medical Faculty and Institute of Tropical Disease, Universitas Airlangga Surabaya


Blood transfusion is an essential part of health care today, however the

transmission of infectious agents by blood and blood products into issues

related to the potential risk of transfusion. Hepatitis C is a disease that can

be transmitted through blood transfusion. Early detection of Hepatitis C

virus in blood donors is needed because given blood donors will then be

transfused to patients who need blood. Examination of anti-HCV antibody

is a frequent checkpoints to detect hepatitis C. However, this method is less

accurate. Therefore, it needs examining more precise and accurate to detect

the presence of hepatitis C. This study was conducted to detect hepatitis C

virus RNA in the blood donor in PMI Tuban, East Java. Methods: A total

of 500 samples of blood donors obtained from PMI in Tuban, East Java,

375 men (75%) and 125 women (25%). Detection of anti-HCV antibody

prevalence in the samples was done by using enzyme-linked

immunosorbent assay (ELISA). Furthermore, the detection of hepatitis C

virus RNA in blood serum which showed positive anti-HCV using a nested

PCR technique. Used three pairs of primer sets targeting the NS5B region /

5'UTR hepatitis C. Further amplification product was visualized in UV on a

2% agarose gel containing ethidium bromide. Anti-HCV positive blood

serum was detected in seven donors (1.4%) using ELISA kit. Of the seven

blood donors testing positive serum anti-HCV, six donor blood serum

showed a hepatitis C virus RNA by using nested PCR.

Keywords: anti-hepatitis C virus, hepatitis C virus, nested-PCR, blood

donor, Indonesia




mailto:[email protected]

19

BMB-2

Regulation of Adipogenesis by Mangosteen Peel Extract and

Xamthones in 3T3-L1 Cells.

Lusiana Darsono1,Wahyu Widowati1*, Jo Suherman1, Khie Khiong1, Seila Arumwardana2,

Merry Afni2, and Hayatun Nufus2

1Faculty of Medicine, Maranatha Christian University, Jl. Prof. drg. Suria Sumantri no 65,

Bandung 40164, Indonesia 2Biomolecular and Biomedical Research Center, Aretha Medika Utama, Jl. Babakan Jeruk 2

No. 9, Bandung 40163, Indonesia

*Correspinding E-mail: [email protected]

One of the risk factors of atherosclerosis is obesity. Fat accumulation and

adipocyte differentiation are associated with the occurrence and

development of obesity. Suppression of adipocyte differentiation is one

potential mechanism for anti-obesity. This study examined the effect of

action mangosteen peel extract (MPE) and xanthone compounds (α-

mangosteen and γ-mangosteen) in 3T3-L1 cells toward expression of

PPARγ, C/EBPα, SCD1, LPL, aP2, adipoQ, and FAS. MPE and xanthones

dose dependently cytotoxic toward 3T3-L1. The concentrations of MPE α-

mangosteen used 0, 25, and 50 µg/ml, meanwhile the concentration of α-

mangosteen used 0, 25, and 50 µM and γ-mangosteen used 0. 50, and 75

µM. The RNA of 3T3-L1 cells was isolated then used for cDNA synthesis.

The expression of PPARγ, C/EBPα, SCD1, LPL, aP2, adipoQ, and FAS

genes were measured using real time quantitative PCR. MPE 50 µg/ml and

γ-mangosteen 50 µM showed actively suppress PPARγ, C/EBPα, SCD1,

LPL, aP2, adipoQ, and FAS gene expression. α-mangosteen 25 µM and α-

mangosteen 50µM couldn’t suppress gene expression of PPARγ and SCD-

1, and also α-mangosteen 50µM failed to reduce aP2 gene expression in

this study. These results showed that MPE and γ-mangosteen have potential

as an anti-adipogenesis and anti–obesity by suppressing gene expression of

PPARγ, C/EBPα, SCD1, LPL, aP2, adipoQ, and FAS in 3T3-L1.

Keywords: Atherosclerosis, adipogenesis, obesity, gene expression,

mangosteen



Malang

BMB-3

Prospects and Problems for Identification of Local Durian from

Ternate Island Using RAPD Markers

Sundari1*, Estri Laras Arumingtyas2 , Luchman Hakim2 and Rodiyati Azrianingsih2

1Departement Biology education, Khairun University of Ternate North Maluku,Indonesia

2 Departement Biology of Brawijaya University, Malang Indonesia

*Coresponding E-mail: [email protected]

There are many local names for durian on the Ternate Island. The giving of

local names were based on morphological variation, the owner's name and

the address where the durians were grown. Thereby, to determine the

taxonomic status of the local durian, the analysis of morphological and

molecular diversity were required. The objective of this study was to

identify the taxonomic status of local durian on the Ternate Island using

RAPD marker. Six primer RAPD were used to identify the genetic

diversity of 27 variants of local durian on the Ternate Island. The analysis

result of polymorphism showed that all primer RAPD effectively

demonstrated polymorphism with the value of 100%. RAPD marker was

effective in detecting genetic diversity (Polymorphism), but it was unable

to show the level of taxa in detail, so application of more advanced

technology markers for barcode DNA was necessary.

Keywords: local durian, Ternate island, RAPD, identification





21

BMB-4

Baculovirus Surface Display using Infuenza Neuraminidase (NA)

Transmembrane Anchor

Irisa Trianti1, Saengchai Akeprathumchai1, Phenjun Mekvichitsaeng2 and Kanokwan

Poomputsa1,*

1 Biotechnology Program, School of Bioresourches and Technology, King Mongkut’s

University of Technology Thonburi, Bangkok, Thailand, 10150 2 Pilot Plant Development and Training Institute, King Mongkut’s University of Technology

Thonburi (Bangkuntien), Bangkok, Thailand, 10150

*Corresponding Email: [email protected]

Baculovirus surface display has been employed as an excellent tools for

presentation of foreign peptides and proteins on budded virus surface with

native conformation, functions and immunogenicity. A baculovirus major

envelope protein, gp64, or a capsid protein, vp39 are generally used as

fusion partners to display polypeptides on the surface of virions.

Alternatively, a membrane anchoring domain of vesicular stomatitis virus

G protein (VSV-G) is also used. In this study, an influenza neuraminidase

(NA) was proposed as an alternative membrane anchor for the display of

Angiotensin II (AngII), DRVYIHPFHL, peptides. The AngII peptides were

inserted into the modified NA, with deleted amino acid residue number 60-

67, and integrated into baculovirus genome to generate a recombinant

baculovirus expressing the recombinant NA fusion-AngII peptides. These

peptides were found to express and translocated on the membrane of the

baculovirus infected insect cell (Sf9 cell) as detected by

Immunocytochemistry using anti-AngII monoclonal antibody. The

conformation of NA on baculovirus surface was not affected by the

deletion, as the 55 kDa band of NA can be detected from Western Blotting

analysis by specific anti-NA monoclonal antibody. In addition, the same

protein was also detected by anti-AngII antibody indicating that the AngII

peptides had been successfully fused with the recombinant NA.

Interestingly, electron microscopy analysis demonstrated that not only the

recombinant baculovirus displaying AngII peptides were generated by

infected insect cells, but also the NA virus-like-particles displaying AngII

peptides.



Malang

Keywords: Baculovirus surface display, Neuraminidase, AngII peptides,

Virus like particles.

BMB-5

Potential Broccoli (Brassica oleracea L.) As Agent Proliferative NK

Cells In Role for the Prevention and Treatment of Cancer

Diana Krisanti Jasaputra1, Khie Khiong1 and Wahyu Widowati1

1 Faculty of Medicine, Maranatha Christian University, Bandung, Indonesia.


Cancer is a disease that causes death. The cancer cells derived from normal

cells that mutate. The immune system plays an important role in malignant

disease. NK cells are non-specific lymphocytes that have the NKG2D

receptor and can kill cancer cells. Agents which increase proliferation of

NK cells can prevent and treat cancer. Broccoli contains sulforaphane has

been shown to have anticancer effects. The mechanism of anticancer agents

can be through an increase in the activity of the immune system, including

increasing proliferation of NK cells that have a role in providing a

protective effect against cancer. The aim of this study was to determine the

effect of broccoli in enhancing the proliferation of NK cells that will be

useful to prevent and treat cancer. This study is an experimental research

laboratory that in vitro using a cell line that NK cell, which gets treated

with the broccoli. Data were measured NK cell proliferation. The test

material used was dried broccoli in the form of powder dissolved in certain

concentrations and given to each cell suspension with a density of 10.000

cells NK cells / cm2. The results of the study after incubated at 37 degrees

C, 5% CO2, for 24 hours, that treat by broccoli with a concentration of 25

ug / ml, 12.5 pg / ml, 6.25 ug / ml, 3.125 mg / ml, and 0 ug / ml

respectively 11,199 cells / cm2, 9,421 cells / cm

2, 9,248 cells / cm

2, 8,308

cells / cm2, and 1,620 cells / cm

2. Conclusion of the study shows that

broccoli increases the number of NK cells is dose dependent, meaning that

with increasing doses of increasing the number of NK cells. This indicates

that broccoli can boost the number of NK cells can be used to prevent and

treat cancer.

Keywords: broccoli, Cancer, NK cells.




23

BMB-6

Inhibition of Pericyte Loss through Ang-2/Tie-2 Signaling Pathway

on Diabetic Rats

Nadia Artha Dewi 1,2*, Hidayat Sujuti 3, Djoko W. Soeatmadji 4,

M.Aris Widodo 5, Aulanni'am 6

1 Department of Ophthalmology - Vitreoretinal subdivision, Faculty of Medicine, Brawijaya

University, Malang, Indonesia. 2 Post Graduate Programme Faculty of Medicine, Brawijaya University, Malang, Indonesia.

3 Department of Biochemistry, Faculty of Medicine, Brawijaya University, Malang,

Indonesia. 4 Department of Endocrinology, Faculty of Medicine, Brawijaya University, Malang,

Indonesia 5 Department of Pharmacology, Faculty of Medicine, Brawijaya University, Malang,

Indonesia 6 Department of Biochemistry, Faculty of Science, Brawijaya University, Malang,

Indonesia.

* Corresponding E-mail: [email protected]

Pericyte loss is the earliest morphological changes in the diabetic retina

which will further affects instability of retinal vasculature. The mechanism

of pericyte loss remain controversial, but recent studies shows that

Angiopoietin-2 have strong correlatted mechanism due to migration of

pericyte. Increase expression of Ang-2 due to hyperglicemia condition will

result in phosphorylation causing cell migration. Angiopoietin regulates

pathways that mediated cellular motility through its receptor, Tie-2, with

activate Phosphotidylinositol 3 kinase (PI3K) and through MAPK

pathways. . The role of Ang-2 in Tie-2 phosporylation is complicated,

depend on the stimuli and the cells. In this study we give intravitreal

injection of Tie-2 inhibitor to find out the mechanism of pericyte migration

on diabetic rat model observed from Ang-2/ Tie-2 signaling pathway in

order to prevent pericyte loss in diabetic rat model.

Keywords: Ang-2/Tie-2, Diabetic rat, Intravitreal injection, Pericyte loss



Malang

BMB-7

Novel Therapy for Ischemic Stroke: Empowering Bone Marrow Stem

Cell (HSC) using β-glucans from Saccharomyces sp on Rattus

norvegicus Ischemic Stroke Model

Khansa Khairunnisa A2, Arinal Mufidah2, Muhammad Unzila Rafsi3, Dedy Budi

Kurniawan2, Rahmad Dwi Saputra3, Wibi Riawan1

1 Laboratorium Biokimia - Biomolekuler, Fakultas Kedokteran Universitas Brawijaya

2 Program Studi Pendidikan Dokter, Fakultas Kedokteran Universitas Brawijaya 3 Program Studi Farmasi, Fakultas Kedokteran Universitas Brawijaya


Stroke is a disease of acute neurological deficits caused by disorder of the

blood vessels of the brain. In General, a stroke can occur in all age groups.

About 80% case of stroke is ischemic stroke. Recent research shows that

the process of neuro-regeneration and the arrangement of the human central

nervous neuroplasticity continues throughout life. Supported by previous

research on stroke, the cell responds with some neuroprotective

mechanisms to protect nerves from neurodegenerative mechanisms, by the

secretion of neurotropin growth factor. The increase of ischemic stroke

prevalence, encouraging us to develop a new effective treatment strategies

by exploiting existing factors in the brain when the stroke occurs and

implementing the mobilization of stem cells to maximize the neuro-

regeneration. Saccharomyces cerevisiae is an unicellular yeasts that

widespread in nature and the potential strains producing β-glucan. β-glucan

in Saccharomyces cerevisiae has the potential to increase G-CSF

(Granulocyte-colony stimulating factor) which is capable of enhancing the

mobilization of HSCs (Hematopoietic Stem cells) in the bone marrow

which then differentiate into neurons so that it can restore the brain

structure and function. The research was conducted in in vivo mouse

models of ischemic stroke using Middle Cerebral Artery Occlusion

Treatment of groups treated with a solution of β-glucans from

Saccaromyces cerevisiae 18, 36, 72 mg/day kgBW. Mice will be dissected

to check the expression of glutamate, neurotropin-1 (NT-1), CD34 with

immunohistochemical metode. The results of immunohistochemical

appears that the expression of glutamate, with beta-glucan, showed a

decrease, while the expression of NT1 and the number of CD34 cells in the

brain tissue was significantly increased (P <0.05) compared to the positive




25

control group. This indicates that beta-glucan may increase mobilization of

HSCs from bone marrow cells to the damaged brain tissue. Thus, beta-

glucan has the potential to be therapeutic for ischemic stroke through

effective mobilization of HSCs.

Keywords: β-glucan, CD34, Ischemic Stroke, Hematopoietic Stem cells,

NT1



Malang

BMB-8

Effects of Co-culture IL2-induced NK (IL2-NK) and human

Wharton’s Jelly Mesenchymal Stem Cells (hWJMSCs) toward TNF-α,

IFN-γ, Perforin, Granzyme Concentration, Cytotoxic and Apoptotic

Inducing Activity in Breast Cancer Cells

Wahyu Widowati1, Diana Krisanti Jasaputra 1, Khie Khiong 1,4, Sutiman B. Sumitro 2, M.

Aris Widodo 3, Merry Afni 4, Seila Arumwardana 4, Dwi Davidson Rihibiha 4 and Indra

Bachtiar5

1Faculty of Medicine, Maranatha Christian University, Bandung, West Java, Indonesia

2Faculty of Science, Brawijaya University, Malang East Java, Indonesia 3Faculty of Medicine, Brawijaya University, Malang East Java, Indonesia

4Biomolecular and Biomedical Research Center, Aretha Medika Utama, Bandung West

Java, Indonesia 5Stem Cell and Cancer Institute, Jakarta, Indonesia


This study was performed to elucidate effect of conditioned medium (CM)

of co-culture IL2-induced NK (IL2-NK) and human Wharton’s Jelly

Mesenchymal Stem Cells (hWJMSCs) toward breast cancer (BC), NK

cells viability, TNF-α, IFN-γ, perforin, granzyme level, apoptosis and genes

expression of Bax, Bcl-2, and p53. Human recombinant IL-2 was used to

induce hWJMSCs and CM-IL2-hWJMSCs was collected to evaluate its

effect toward BC cells (MCF7) and NK cells. The co-culture method of

IL2-NK and MCF7, IL2-NK and hWJMSCs for evaluating the TNF-α,

IFN-γ, perforin, granzyme level. The measurement of cytotoxic activity of

CM-IL2-hWJMSCs toward MCF7 and NK cells, viability of IL2-NK in

various incubation period, cytotoxic activity of IL-2 NK toward MCF7

cells in various ratio were conducted using 3-(4,5-dimethylthiazol-2-yl)-5-

(3-carboxyme-thoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS)

assay, concentration of TNF-α, IFN-γ, perforin, granzyme using ELISA

assay. Apoptosis was measured using flow cytometry, and apoptotic genes

were measured with RT-PCR. The CM-IL2-hWJMSCs and CM-hWJMSCs

exhibited cytotoxic effect, higher concentration of CM (60%) showed

higher inhibition in MCF7, NK cells. The higher number of NK cells

increased the anticancer activity toward BC cells, the highest cytotoxic

activity was obtained from co-culture of BC and NK cells in ratio of 1:5.

The IL2-induced NK cells proliferation were higher compared to the




27

noninduced cells. Co-culture of NK cells and hWJMSCs, MCF7 cells

produced TNF-α, IFN-γ, perforin, granzyme. IL2-induced NK cells

produce higher level of TNF-α, IFN-γ, perforin, granzyme compared to

IL2-noninduced NK cells. The hWJMSCs and MCF7 cells did not secrete

perforin, granzyme, secreted low level of TNF-α, IFN-γ. Highest level of

TNF-α, IFN-γ, perforin, granzyme were secreted by co-culture of

hWJMSCs, MCF7 cells and NK cells in comparison 1:5. CM-( hWJMSCs

+ IL2-NK) showed highest apoptosis among treatments, as also supported

by increased genes expression of BAX and P53, and decreased BCL-2. The

CM-IL2-hWJMSCs and CM-hWJMSCs have anticancer activity against

BC cells and toxic against NK cells. The IL-2 increase NK cells

proliferation, longer incubation increase NK cells number. Higher cells

number of NK and IL-2 inducer increase secretion level of TNF-α, IFN-γ,

perforin, granzyme both in hWJMSCs and MCF7 cells. CM of co-culture

hWJMSCs and IL2-NK cells induce apoptosis in MCF7, as indicated by

increased BAX and P53, and decreased BCL-2.

Keywords: human Wharton’s Jelly Mesenchymal Stem Cells, Interleukin-

2, Conditioned Medium, Natural Killer, Breast Cancer,

Perforin, Granzyme, Apoptosis



Malang

BMB-9

Isolation and Mercury Sensitivity Test of Bacterias Isolated fron

Waste Disposal in Gold Mining Area in West Sekotong of West

Lombok Region

Atiqa Ulfa1*, Endang Suarsini1 and Mimien Henie Irawati al Muhdhar1

1State University of Malang, Jalan Semarang 5, Malang, East Java, Indonesia


Type B of wild mining might be the source of complicated environmental

problem for the using of mercury. The analysis of mercury on June 2015

showed the mercury level of the waste disposal as much as 4,04-29,88

ppm. High toxicity of mercury is threatened to contaminate the water

source for residents of temporary waste is discharged directly to the yard of

residents. Organic compunds un the waste disposal is a source of nutriens

for microbes. Microbes will break down the compunds into a from that is

more simple and stable so that the levels if contaminants can be reduced.

Enzymatic reaction by bacteria is the key to the implementation process of

gradual transfromation in the management of waste disposal from the

substrase that is generally in the complex molecular structure, into simple

elements. This preliminary study aims to isolate and to know the bacterias

sensitivity to mercury isolated from the temporary shelter of gold mining

waste disposal area in West Sekotong of West Lombok region. The

disposal was taken from three different locations. Bacterias were isolated

and then inoculated on NA-HgCl2 medium. Identification of bacterias is

based on morphological, physiological and biochemical test. The results

showed that four bacterias were successfully isolated, all of the isolated

bacterias were Gram positive bacterias. Only three bacterias showed

motility and another one was non motil bacteria. Sensitivity test showed

that two bacterias were resistant and another two were sensitive.

Keywords: bacteria, mercury, waste disposal.




29

BMB-10

Modulation of Endogenous Stem Cells Markers: STRO-1, CD44,

CD105, CD146 in Periodontal Ligament of Dental Rat Post Traumatic

Avulsion after Aloevera Exposure

Yuli Nugraeni1, Wibi Riawan 2 and Nur Permatasari3

1Doctoral Program in Medical Science Student Interests Biomedical Graduate School of

Medicine, Brawijaya University 2Laboratory of Molecular Biochemistry of Faculty of Medicine, Brawijaya University

3Laboratory of Pharmacology of Faculty of Medicine, Brawijaya University

* Corresponding E-mail:

Heterogeneity and continuous remodeling capability in periodontal ligament

tissues indicates there are progenitor cells that produce specialized cell types, is

known as the periodontal ligament stem cells (PDLSCs). The Periodontal

ligament is a connective tissue that composed mainly fibroblasts. The

regulation and the presence of fibroblasts cells had not been clear. It has been

thought, the original is from stem cells system that can continuously renewing

the cell. The purpose of this study is to determine the distribution of

endogenous periodontal ligament stem cells markers; Stro-1, CD44, CD105

and CD146 in the periodontal ligament tissue of dental Wistar rats post

traumatic avulsion. This research was a true experimental laboratory by post

test randomized controlled group design in animal models. The rats were

performed extracted the incisive teeth of the upper jaw. There were two

groups, control group and the other group with aloe vera administrations. After

post-extraction, each group were divided into four treatment groups based on

time of exposure; 1, 3, 7, and 14 days. After the rat sacrifice, the periodontal

ligament tissues were preserved by immunohistochemistry using specific

antibodies: Stro-1, CD44, CD105 and CD146. Analyzing data is using

statistical analysis with Oneway ANOVA, Tukey HSD, Pearson Correlation

and Regression Testing. The results of this study showed that there were no

significant differences between the control group and the treatment group. The

conclusion of this study shows the distribution of endogenous stem cells to

specific antibody Stro-1 showed the most significant periodontal ligament stem

cells distribution and can be used as a biologic marker.

Keywords: PDL stem cells, aloe vera, Stro-1, CD44, CD105, and CD146



Malang

MVS-1

Therapeutic Effect of Goat Milk Yogurt on Tumor Necrosis Factor

Alpha (TNF-α) Expression and Histopathology Changes of Aorta in

Hypercholesterolemic Rat (Rattus norvegicus)

Chanif Mahdi1*, Masdiana C. Padaga2 and Andhika R. Pangetika2 1. Departement of Chemistry, Faculty of Mathematics and Natural Sciences, University of

Brawijaya. 2.Program Study of Veterinary Medicine, Veterinary Medicine Program,

University of Brawijaya


Hypercholesterolemia is a risk factor for cardiovascular disease which

causes the formation of oxidized LDL and triggers an inflammatory

reaction, characterized by the release of cytokines TNF-α in the aorta. Goat

milk yogurt containing lactic acid bacteria (LAB), bioactive peptides and

antioxidants be an alternative therapy of hypercholesterolemia to reduce

blood cholesterol levels. The purpose of this study was to determine the

effect of goat milk yogurt therapy for hypercholesterolemia in terms of the

expression of TNF-α and histopathological picture of the aorta. This study

used 20 male rats Rattus norvegicus, Wistar strain were divided into 5

groups: control group (A), hypercholesterolemia group (B),

hypercholesterolemia group and therapeutic dose of 300 mg / kg (C),

hypercholesterolemia group and therapeutic dose of 600 mg / kg (D), as

well as hypercholesterolemia and therapeutic dose group of 900 mg / kg

(E). Rat model of hypercholesterolemia got hypercholesterolemia diet for

14 days and goat milk yogurt therapy for 28 days. The results showed

highly significant differences for each treatment group (p <0.01), which

contained an average decrease in the expression of TNF-α by 61.14% (C);

37.54% (D); and 17.89% (E). Besides picture aortic histopathology showed

that there was no improvement of endothelial desquamation, reduced lipid

accumulation and infiltration of inflammatory cells in the aorta. The

conclusion from this research that therapy goat milk yogurt can decrease

the expression of TNF-α and improve picture rat aortic histopathology

hypercholesterolemia models, with the most effective dose is a dose of 900

mg / kg.

Keywords: hypercholesterolemia, goat milk yogurt, TNF-α, histopathology

aorta





31

MVS-2

Goat Milk Yogurt Reduce Systolic Blood Pressure (SBP) and Repair

Brain Damage in DOCA - SALT Hypertensive Rat

Masdiana Padaga1*, Aulanni’am2

and Ganendra Awang Kristandya1

1 Faculty of Veterinary Medicine, Brawijaya University,

Malang, 65145, Indonesia. 2Department of Biochemistry, Faculty of life Sciences, Brawijaya University,

Malang, 65145, Indonesia.


Goat milk yogurt fermented with starter containing Lactobacillus

bulgaricus, L. acidophilus and Streptococcus thermophilus generated

bioactive peptides with blood pressure lowering activity. Antihypertensive

activity of the yogurt in Deoxycorticosterone acetate (DOCA)–salt induced

hypertensive rats (Rattus norvegicus) strain Wistar were studied. The rats

were orally administrated with the fermented goat milk in daily dose of 300

and 600 mg/kg body weight for 4 weeks. The effect of yogurt treatment on

Systolic blood pressure (SBP), histopathology and Malondialdehyde

(MDA) level of hypertensive rats brain were evaluated. The goat milk

yogurt contained 22.80% of protein and the yogurt (8 %, w/v) showed

21.70 % ACE-I activity. Daily administration of 300 and 600 mg/kg bw of

the yogurt prevented the increase in SBP. Treatment of the hypertensive

rats with the yogurt suppressed endothelial damage as well as reduced

hemorrhage. The current study suggests that goat milk yoghurt treatment

was capable of reducing MDA levels and repair histopathological

alterations in brain of DOCA-salt hypertensive rats.

Keywords: Goat milk, DOCA–salt, yogurt, hypertension, endothelial

damage.



Malang

MVS-3

The Success Rate of Early Sexing Canaries through PCR

Mudawamah1*, M.Z. Fadli1 and G Ciptadi2

1Islamic University of Malang, MT Haryono 193, Malang, 65155, Indonesia

2Brawijaya University of Malang, Veteran, Malang, 65145, Indonesia

*Corresponding Email: [email protected]

Determining of birds sex, including canaries were very important in

supporting of breeding as well as maintenance management. The purpose

of this study was to conduct early sexing through molecular analysis of

polymerase chain reaction (PCR). With the development of this method

was expected to do sexing when the birds began to grow feather more than

10 days of age. DNA samples were taken from the feather with specific

primers for sexing. Preliminary results showed PCR was able to identify

the sex of the bird through the feather samples with a success rate of 58%.

Therefore PCR method could be developed as early sexing canary

Keywords: early sexing, canary, PCR




33

MVS-4

Detection of Arsenic in Cattle’s Blood Samples using Atomic

Absorption Spectrophotometer

Nemay A Ndaong1 and Annytha Detha2*

1 Departement of Pharmacology, Faculty of Veterinary Medicine, Nusa Cendana University,

Indonesia

Jl. Adi Sucipto, Penfui. Kupang-East Nusa Tenggara, Indonesia 2 Department of Public health Veterinary Faculty of Veterinary Medicine, Nusa Cendana

University, Indonesia

Jl. Adi Sucipto, Penfui. Kupang-East Nusa Tenggara, Indonesia


The aim of this study was to detection of Arsenic in blood of cattle in the

landfill. A sample of 12 cattle were classified into three age groups, the

groups of 0 to 1.5 years, 1.5 to 3 years and over 3 years. Cattle’s blood

samples taken for arsenic detection using Atomic Absorption

Spectrophotometry variant AA240FS. The results show that there are 8

samples that have a high content of arsenic and exceeding the tolerance

limit. Lead content varied from 0.2860 to 0.9652 ug / ml. The conclusion of

this study also showed that blood samples of cattle aged three years old

have the highest arsenic.

Keywords: Arsenic, cattle, blood, landfill



Malang

GHG-1

A novel mutation of iduronate-2-sulfatase gene in an adult patient with

severe Hunter syndrome

Antonius Christianto1 and Tetsuya Inazu1

1 Department of Pharmacy, College of Pharmaceutical Sciences, Ritsumeikan University,

Kusatsu, Shiga


Mucopolysaccharidosis II (Hunter syndrome), a lysosomal storage disorder

caused by a deficiency of iduronate-2-sulfatase (IDS), has variable clinical

phenotypes. Total by nearly 400 different mutations have been identified in

IDS gene from patients with Hunter syndrome. Herein, we report a patient

who has a novel mutation in IDS gene with a severe clinical phenotype and

effect of enzyme replacement therapy. Gene mutational analysis was

conduct using genomic DNA extracted from human blood and continued

with direct DNA sequence. Patient was administered with Idursulfase

(Elaprase®) for enzyme replacement therapy (ERT). Level of urinary

glycosaminoglycan (GAG) was measured before and after treatment.

Genetic analysis of the IDS gene revealed a novel 1-bp deletion in position

c.1053T in exon 8 and resulting in a frameshift with a premature stop

codon. Predicted product consists of 358 amino acids which lacks

approximately 30% c-terminal portion of full IDS including several N-

glycosylation sites. ERT using Elaprase® was conducted to the patient.

Effect of this recombinant enzyme are improved hepatosplenomegaly,

white blood cells and platelets number, and decreased the level of urinary

glycosaminoglycan (GAG). ERT was proved to be effective in patient with

severe type of Hunter syndrome.

Keywords: deletion mutation, glycosaminoglycan, Iduronate-2-sulfatase

gene, Idursulfase enzyme replacement therapy.




35

GHG-2

Profiling Intermarriage of Tionghoa Ethnic from Two Family Based On

Antropometrics and 13 Codis DNA

Ananda Aqmarina Chaerani1

1 Biology Department, Mathematic and Natural Science Faculty, Brawijaya Unniversity


Tionghoa ethnic is one of migrant ethnic which is a largest population than other

migrant ethnic in Indonesia. Now day, intermarriage Tionghoa with Javanese,

Madurese, Sundanese, or other are a much, however the profile of DNA fingerprint

still unknown. The aim of this study is to identify anthropometrics specific

character and also the similarity and diversity of DNA fingerprint profile of

intermarriage Tionghoa family using 13 CODIS DNA. Anthropometric

measurement is done by measure length of stature, upper extremities, lower

extremities, palm, and fingers. The anthropometric will be measure before taking

the blood samples. DNA samples collect from blood of three generations from two

Chinese families. DNA extraction from blood samples extracted using salting-out

technique. Quantitative measurement of DNA measure by UV-Vis-

spectrophotometry and qualitative detect by 1% agarose gel electrophoresis. PCR

perform using 13 CODIS marker, TPOX, D3S1358, FGA, D5S818, CSF1PO,

D7S820, D8S1179, TH01, VWA, D13S317, D16S539, D18S51, D21S11, and

AMEL. The DNA-PCR run using 8% polyacrilamide gel electrophoresis. The

DNA-PCR profile visualize with Chemidoc Gel Imaging (Bio-rad) and analyze

using Quantity One software to analyze the similarity, genetic variability, and

alleles pattern. Result showed the general specific DNA band identified using

D7S820 marker. New variance in 1st family showed using D16S539, D21S11 and

D8S1179 markers, while in 2nd

family remain unidentified. Anthropometric

analysis showed 1st family had longer upper extremities than lower extremities,

while 2nd

family had longer lower extremities than upper extremities.

Keywords: 13 CODIS, DNA fingerprinting, Tionghoa ethnic



Malang

GHG-3

Determining the Genetic Similarities and Variability of Javanese and

Arab Ethnic Families with DNA Fingerprint in Malang East Java Indonesia

Nila Kartika Sari1, Eriko Prawestiningtyas2 and Fatchiyah3

1 Departement of Biology, Mathematic & Natural Sciences, IKIP Budi Utomo Malang

East Java Indonesia 2 Laboratory of Forensic Medicine, Medical Faculty, Brawijaya University Malang East

Java Indonesia 3 Departement of Biology, Mathematic & Natural Sciences Faculty, Brawijaya University

Malang East Java Indonesia


More than one-third of human genome consists of repetitive sequence

region (Repeat Area) which consist of Minisatellite or Variant Number Of

Tandem Repeats (VNTR) and Microsatellite or Short Tandem Repeat

(STR). Based on its short allele range STR can be used for the paternity

testing study of genetics disease, molecular archeology, as well as in

forensic crime cases. The aim of this study is to identify Javanese – Arab

Ethnic DNA fingerprint in determining the similarities and genetic variability

Materials and methods to accomplish this, we used human white blood cell

from three generations of three family consists of: (1) grandmother-mother,

father-daughter, (2) grandfather-mother, father-daughter, (3) grandfather,

grandmother–mother, father-son DNA blood samples were Isolated by salting

out, furthermore PCR amplification used by applying 13 CODIS which

consists of TPOX, D3S1358, FGA, D5S818, CSF1PO, D7S820, D8S1179,

TH01, VWA, D13S317, D16S539, D18S51, D21S11 and amelogenin, and

then it was visualized by 8% polyacrylamid gel. The Fingerprint profile

was visualized by 8% polyacrylamide gel and took the picture by

ChemiDoc gel Imaging and measure the intensity band pattern by Quantity

One software. Variations in the pattern of DNA bands were analyzed using

the program GENEPOP software package version 4.2 that will be obtained

allele frequencies, heterozygosity, and allele migration. Based on

identification, this result showed that the genetic variability and

heterozygote allele increasing markers from the first generation to the next

generation with paternity testing from each family were matched. Based on

the results of the analysis heterozygosity values, population III (93.8461%)

have higher heterozygosity values compared with the population I





37

(88.4615%) and II (76.9230%) and migration of alleles 0.341373%. The

percentage of the migration though minor allele had occurred Breeding

populations between Java to the Arab population thereby increasing the

average value of heterozygosity in each population. The conclusion of this

study, marriage variability. Patterns of heterozygous alleles with values

based on heterozygosity, number of alleles at D21S11, VWA and THO1

can be recommended as a molecular marker for the identification of genetic

variation.

Keywords: 13 CODIS, DNA fingerprint, Javanese – Arab Ethnics.



Malang

BE-1

Treatment Planning System Development Opportunities Using 3D

Printing in Indonesia

Thareq Barasabha1*and Yati Hardiyanti2

1Medical Record and Health Information Department, Politeknik Piksi Ganesha, Bandung,

Jawa Barat, 40274, Indonesia 2Physics Department, Institut Teknologi Bandung, Bandung, Jawa Barat, 40132, Indonesia


Along with the increasing life expectancy, the chances of the cancer

incidence is also increasing. Referral hospitals in the capital cities of some

provinces in Indonesia need to prepare themselves to form a high-quality

radiotherapy center. It does not only require advanced radiotherapy

equipment, but one of the things that must also be prepared by these

radiotherapy centers is the development of Treatment Planning System

(TPS). With the rapid development of three dimensional (3D) printing and

3D printing services are available in major cities in Indonesia, we think that

3D printing can be an interesting supporting equipment for the

development of TPS on radiotherapy centers in Indonesia.

Keywords: 3D printing, radiotherapy, treatment planning system.




39

BBS-1

The Role of Polysaccharide Krestine (PSK) as an Immunomodulator to

the Number of T Regulatory Cell Cd4+Cd25+Foxp3

+ In Rats with

Arthritis Adjuvant

Diah Purwaningsari1

1Department of Histology (Lecturer) Hang Tuah University Medical Faculty


Regulatory T cells are cells that play a role in maintaining the balance of

the immune, preventing autoimmune, reducing inflammation and

minimizing tissue damage. Its main function is to inhibit the function of

antigen-presenting cells (APC) and effector T cells (Teff cells). Reducing

the activity of Treg cells may be associated with autoimmune diseases in

humans such as Rheumatoid Arthritis (RA). Treg cells express on their

surface CD4 and CD25, but more definitive identified by the master of their

regulatory transcription expression ie FoxP3. Rheumatoid Arthritis (| RA)

even if only 1% of the world, but it is a chronic autoimmune inflammatory

disease that can lead to disability and decreased life expectancy. The use of

experimental animals to learn more about the disease has long been used is

called adjuvant arthritis (AA). AA can be induced in susceptible strains of

mice, with a single subcutaneous injection H37Ra Mycobacterium

tuberculosis (Mtb) inactivated by heating and dissolved in oils, one of

which is known as Complete Freund's Adjuvant (CFA). Utilization of

Indonesian natural materials, Coriolus versicolor mushrooms, containing

Polysaccharide Krestin (PSK) with β-glucan bond is expected to be an

alternative therapy for the disease. The active ingredient that functions as

an immunomodulator in PSK is a β-glucan, which will be recognized by

toll like receptor (TLR) that is specific to macrophages. Furthermore, β-

glucan will induce the development of regulatory T cells CD4 + CD25 +

FoxP3 +. This study uses three groups of male rats Rattus norvegicus

weighing 200-250 gr, each containing 11 rats. One negative control group,

one positive control group, in the CFA injection, one group treated with

CFA injection and PSK given orally by stomach sonde dose of 50 mg / kg

for 3 weeks. We observe the size of the animal thick hind legs to look at the

volume and checks the blood flowcytometry try to count the number of

regulatory T cells CD4 + CD25 + FoxP3 +. Results: PSK as an



Malang

immunomodulator with a dose of 50 mg/ kg body weight may increase the

number of T cells Regulatory CD4+ CD25

+ FoxP3

+ in the blood of white

rats with adjuvant arthritis and can inhibit the increase in foot volume as a

result of inflammation.

Keywords: PSK, β-glucan, immunomodulator, CD4+CD25

+FoxP3

+

Regulatory T cell , paw volume




41

BBS-2

Apgar scores of breech presentation: Which one is better, vaginal or

cesarean delivery?

Jonny karunia Fajar1*, Mohd Andalas2 and Harapan Harapan1

1 Medical Research Unit, School of Medicine, University of Syiah Kuala, Banda Aceh,

Indonesia 2 Obstetrics & Gynaecology Department, dr. Zainoel Abidin Teaching Hospital, Banda

Aceh, Indonesia


Mode of delivery for breech presentation is a controversy. Several studies

have reported the advantages and disadvantages of each delivery mode for

breech presentation. The aim of this study was to compare breech

presentation`s Apgar scores between vaginal and cesarean delivery. This

study was retrospective, conducted in dr. Zainoel Abidin Teaching Hospital

during January 2010 – Desember 2012. The following data were extracted

from medical record. i.e: mode of delivery for breech presentation, apgar

score in first minute, and apgar score in fifth minute. Data of odds ratio

(OR) and 95% confidence interval (95%CI) were analyzed using double

logistic regression. RESULTS: A total of 153 (3.9%) breech presentations

of 5,252 deliveries between January 2010 and December 2012 were

analyzed for this study. There were 26% (52 cases) vaginal and 74% (153

cases) cesarean delivery. Our results showed that mode of delivery for

breech presentation had the significant association with apgar score in first

minute (OR95%CI = 4.193 [1.636 – 10.746], p = 0.003) and fifth minute

(OR95%CI = 7.977 [1.384 – 45.987], p = 0.020). In summary, there was

significant association between Apgar scores of breech presentation and

mode of delivery. Our results suggested that cesarean delivery for breech

presentation had better outcome.

Keywords: Apgar score, Breech presentation, cesarean delivery, vaginal

delivery.



Malang

BBS-3

Red Dragon Fruit Extract Reduces Liver Malondialdehyde Level

Induced By High Dose Paracetamol In Male Wistar Rats

Putri Dariyanti Wijaya1* and Sulistiana Prabowo1

1 Faculty of Medicine, Hang Tuah University, Surabaya, Indonesia


The administration of high dose paracetamol increases the level of N-

acetyl-benzoquinoneimine (NAPQI). This metabolite has a reactive

characteristic causes oxidative stress and increases liver malondialdehyde

(MDA). This research aimed to determine the effect of red dragon fruit

extract in reducing the liver MDA level induced by high dose paracetamol

in male Wistar rats (Rattus novergicus). Male Wistar rats (n=24) were fed

standard diet and divided into 3 groups. The first group were fed with

standard diet only, the second group received additional 1750 mg/kgBW

paracetamol orally on day 9, and the last group received the same dose of

paracetamol and 300 mg/kgBW/day red dragon fruit extract orally every

day for 10 days. On the tenth day, all rats were sacrificed and the liver

MDA level was examined using the thiobarbituric acid method. Using

ANOVA test the liver MDA level of the group that received high dose

paracetamol (𝑥 =1060.75±117.949nmol/g) was significantly higher

(p=0.005) compared to the group that was fed with standard diet

(𝑥 64. 5±197.409nmol/g). The level of liver MDA was lower in the

group that received high dose paracetamol and red dragon fruit extract

(𝑥 100 .00±230.604nmol/g) compared to the group that received only

paracetamol but the difference was not statistically significant (p=0.574). It

can be concluded that administration of high dose paracetamol increased

liver MDA level significantly, while the administration of red dragon fruit

extract tend to reduce the liver MDA level as this fruit contains flavonoid,

betacarotene, vitamin C and vitamin E as antioxidant.

Keywords: Hylocereus polyrhizius, malondialdehyde, Paracetamol.




43

BBS-4

The Effect of Super Red Dragon Fruit Skin Extract to Increased SGPT

and SGOT Activities Induced by High Dose Paracetamolin Male

Wistar Rats

Nieke Tussina Widhowati1 and Fitri Handajani1*

1 Faculty of Medicine, Hang Tuah University, Surabaya


High dose paracetamol increase N-acetyl-p-benzoquinone imine (NAPQI)

metabolite which is a free radical and toxic to liver and associated with

increased SGPT and SGOT activity. The super red dragon fruit (Hylocereus

costaricensis) skin extract has antocyanin which may quench free radicals.

The aim of this study was to determine the effect of super red dragon fruit

skin extract to SGPT and SGOT activity induced by high dose paracetamol

in male Wistar rats (Rattus norvegicus). We used 24 male Wistar rats

divided into three groups. First group: Rats were given standard food.

Second group: Rats were given additional paracetamol 1750 mg/kgBW per

oral on day 9. Thirdgroup: Rats were given paracetamol 1750 mg/kgBW

per oral on day 9 and 217.73 mg/kg BW Hylocereus costariensis skin

extract per oral every day for 10 days. At the end ofexperiment, all rats

were sacrificed and SGPT and SGOT activity were measured using

spectrophotometry method. Mann-Whitney test showed that there was

significantly higher SGPT(p=0.027) and SGOT (p=0.001) activity in the

second group compared to the first group. There was significantly lower

level of SGOT in the third group compared to the second group. There was

no significant difference in SGPT activity (p=0.141) between the second

and the third group. High dose paracetamol increased SGPT and SGOT

activity significantly. Super red dragon fruit skin extract decreased

paracetamol induced SGPT activity significantly and tend to decrease

SGOT activity possibly due to the antocyanin antioxidant content.

Keywords: Hylocereus costaricensis, paracetamol SGOT, SGPT.



Malang

BBS-5

The Effect of Super Red Dragon Fruit Skin Extract to Serum Catalase

Activity Induced by High Doses Paracetamol in Male Wistar Rats

Ivana Onggowarsito1 and Fitri Handajani1*

1Medical Faculty, Hang Tuah University, Surabaya


Paracetamol can cause side effects if taken in high doses. Free radical N-

acetyl-p-benzoquinone imine (NAPQI) metabolites, a result of the drug

metabolism cause oxidative stress which decrease serum catalase.

Additional exogenous antioxidants is required to anticipate the effects of

these free radicals. Super red dragon fruit (Hylocereus costaricensis) skin

contain antioxidants such as antocyanin. This study aim to determine the

effect of super red dragon fruit skin extract to serum catalase activity of

male Wistar rats induced by high dose paracetamol. This study was an

experimental research using post-test only control group design. We used

24 male Wistar rats divided into three groups. All groups received standard

food diet. The first group received the standard diet only, while the second

group received additional high dose paracetamol (1750 mg/kg BW) per oral

on day 9. The third group received additional high dose paracetamol (1750

mg/kg BW) per oral on day 9 and 218 mg /kg BW per oral super red

dragon fruit skin extract every day for 10 days. Mann Whitney test showed

there was significantly lower level of serum catalase in the second group

compared to the first group (p=0.045). The third group has significantly

higher level of serum catalase compared to the second group (p= 0.003).

High dose paracetamol significantly decreased serum catalase activity in

male Wistar rats. Super red dragon fruit skin extract increased serum

catalase activity in male Wistar rats induced by high dose paracetamol, it

may because of antocyanin which have antioxidant activity.

Keywords: Catalase, Hylocereus costaricensis, Paracetamol.




45

BBS-6

The Effect of Ethanol Green Tea (Camellia sinensis) Leaves Extract in

Zebrafish (Danio rerio) Embryo to Heart Malformations, Reduced of

Body Length, and Hypopigmentation on Larval Stage

Nurdiana1, Subandi Reksohusodo2, Mukhamad Nooryanto3, I Wayan Arsana Wiyasa3,

Siswanto4, Sabatina Windyaningrum5, Made Pradnyawati Chania5, Fadilatul Muzayyanah5

1Lecturer, Pharmacology Laboratory, Faculty of Medicine, University of Brawijaya,

Malang, Indonesia

2Lecturer, Bachelor of Midwifery, Faculty of Medicine, University of Brawijaya, Malang,

Indonesia 3Lecturer, Department of Obstetrics and Gynecology, Saiful Anwar General Hospital,

Malang, Indonesia

4Lecturer, Public Health and Preventive Medicine, Faculty of Medicine, University of

Brawijaya, Malang, Indonesia

5Student, Bachelor of Midwifery, Faculty of Medicine, University of Brawijaya, Malang,

Indonesia


The dominant contents of ethanol extract of green tea / Camellia sinensis

(EGT) are EGCG (epigallocatecin gallate), quercetin, and kaempferol.

EGCG is a competitive inhibitor of dihydrofolate reductase (DHFR)

enzyme in folate-mediated one-carbon metabolism. This mechanicm can

cause an abnormal balance of folic acid in pregnant women. It can interrupt

the embryogenesis, which related with anencephaly and spina bifida.

Meanwhile, EGCG, quercetin, and kaempferol are known to be inhibitors

of tyrosinase enzyme which catalyzes the synthesis of melanin in

melanocytes. Tyrosinase enzyme deficiency can cause oculocutaneous

albinism type 1. The purpose of this study is to determine the effect of EGT

in cardiac malformations, body length, and hypopigmentation of Zebrafish

(Danio rerio) Larvae. Experimental study using randomized post-test only

controlled group design conducted onto zebrafish embryos. The samples

used were devided into four groups: K (Control group); P1, P2, P3 (each

treated with EGT concentration of 125, 250, and 500 ppm). Zebrafish

embryos are given exposure to the ethanol green tea extract at 2, 24, and 48

hpf. The observation made on the larval stages of 72 hpf using OPTILAB

software. The One-Way ANOVA test result showed reduction of heart rate

(p=0.003) and cardiac morphological abnormalities (p=0.000), it also

reduced body length (p<0.000), and caused hypopigmentation (p<0.000)



Malang

with significant differences. In conclusion, exposure of EGT leaves extract

in embryo phase caused cardiac malformations, reduced of body length,

and hypopigmentation in zebrafish larvae. This research can be considered

on determining if it’s safe to consume green tea during pre-conception or

early pregnancy periode.

Keywords: Camellia sinensis, cardiac malformations, EGCG,

hypopigmentation, reduce body length, zebrafish.




47

BBS-7

The Effect of Neem Leaf Extract to Liver Malondialdehyde Level

Induced by High Dose Paracetamol in Male Wistar Rats

Shabrina Mutiara Alimah1*, Nita Pranitasari1 and Sulistiana Prabowo1

1Faculty of Medicine, Hang Tuah University, Surabaya Indonesia

*Corresponding email: [email protected]

High dose paracetamol increases the production of N-acetyl-p-

benzoquinoneimine (NAPQI). This metabolite has reactive characteristics

and causes oxydative stress that increases lipid peroxydation and liver

malondialdehyde (MDA) level. This research aimed to determine the effect

of neem leaf extract (Azadirachta indica Juss.) in reducing liver MDA level

induced by high dose paracetamol in male Wistar rats (Rattus novergicus).

Three groups of eight male Wistar rats were fed standard diet. The first

group received standard diet only, the second group received an additional

single dose of paracetamol (1750 mg/kgBW) on the ninth day, and the last

group received the same dose of paracetamol and 500 mg/kgBW neem leaf

extract for ten days. On the tenth day, the rats were euthanized and the liver

MDA level was measured using thiobarbituric acid method. Data analysis

using Kruskal-Wallis test showed that liver MDA level of the group that

received high dose paracetamol ( 2043.50+168.17nmol/g) was

significantly higher (p=0.036) compared to the group that received

standard diet ( 1199.50+677.98nmol/g). Liver MDA level of the group

that received high dose paracetamol and neem leaf extract (

1703.25+199.69nmol/g) was significantly lower (p=0.009) compared to the

group that received high dose paracetamol ( 2043.50+168.17nmol/g). In

conclusion the administration of high dose paracetamol significantly

increased liver MDA level in Wistar rats and neem leaf extract significantly

reduced paracetamol induced liver MDA level as neem leaf contains

flavonoid and limonoid that function as antioxidant.

Keywords : Azadirachta indica Juss., malondialdehyde, paracetamol.



Malang

BBS-8

Innovation Therapeutic uses Centella asiatica extract form Phytosome

with Time Release Technology Composite Membrane Application in

Modulation of Endogenous Neural Stem Cell Rat Model Traumatic

Brain Injury

Rizkha Farida2, Erlin Aditia Purmitasari3, Ika Dewi Soraya2, Lilin Andreas Vitanea3, Wahyu

Dwi Nugroho2, Wibi Riawan1

1. Laboratorium Biokimia - Biomolekuler, Fakultas Kedokteran Universitas Brawijaya

2. Program Studi Pendidikan Dokter, Fakultas Kedokteran Universitas Brawijaya

3. Program Studi Farmasi, Fakultas Kedokteran Universitas Brawijaya


Traumatic Brain Injury (TBI) is an injury to the brain due to a serious

collision. After TBI will inflammatory reaction as a defense of nervous

tissue that is characterized by elevated TNF-α. Central Nervous System

adult mammalian contains two areas neurogenic: zona subgranular (SGZ)

of the hippocampus and the zone subventricular (SVZ) of the lateral

ventricle (Alvarez-Buylla and Lim, 2004; Cameron et al, 1993; Gould et

al., 1998; Kuhn et al, 1996). TBI can activate stem cells, which may reflect

the induction of innate mechanisms repair and plasticity of the brain injury,

the study by Gao et al. 2009, proving that neurogenesis can be caused by

TBI, and an increase in cell proliferation was observed in DG and SVZ.

Sox2 appears terekpersi the NPCs from the subventricular zone and in the

layer of the dentate gyrus of the hippocampus subgranular. NPCs marker

nestin and glial marker GFAP (glial fibrillary acid protein), are

characterized by their ability for self-renewal and their potential in the

regeneration of neurons and glial cells. The process of neurogenesis can be

modulated through involving Brain-derived neurotrophic factor (BDNF).

Centella asiatica containing asiaticoside has the ability to inhibit

inflammation and regulate the levels of intracellular Ca2 +, thereby

increasing BDNF. However, the phytochemical components in the extract

of Centella tend polar that need to be formulated in the form Phytosome.

To reduce the toxicity of required local dosage formulations in the form of

a hydrogel with time release system, so that the process of homeostasis

occurs and inhibit inflammatory hyperactivation. The purpose of this study

prove the technology time relase composite membrane applications

Phytosome Centella asiatica extract in inhibiting TNF-α and modulating




49

endogenous NSCs through increased BDNF, SOX2, and S100B rat model

of TBI. Rats are grouped into 4 groups: control (-), control (+), the

treatment group were given hydrogel, and the treatment group were given

hydrogel Phytosome extract of Centella asiatica 90 mg / kg. Measurement

of the expression of TNF-α, BDNF, SOX2, and S100B using

immunohistochemistry (CPI). Data obtained in nertuk number and

description of each protein expression. The analysis was performed to

examine differences in the expression of each parameter using multivariate

ANOVA. The results showed that the hydrogels without Phytosome no

effect on the inhibition of TNF-α in spite of having an influence on the

increase in BDNF and SOX2, hydrogel without Phytosome not affect

maturation of nerve cells that are marked S100B so that the hydrogel

without Phytosome only serve as drug carriers. Meanwhile, administration

of hydrogels Phytosome affect inhibition of TNF-α, starting on day 3, the

increase in BDNF, increase SOX2, and S100B, starting on day 1 as a sign

of neurogenesis. Conclusion of research is gotu kola extract Phytosome

therapy with time release technology applications of composite membrane

is able to inhibit TNF-α, modulate endogenous NSCs through an increase in

BDNF, SOX2, and S100B in mice models of TBI.

Keywords: Centella asiatica, Endogenous NSC, Hydrogel, Phytosome,

TBI



Malang

P-01

Selective Cytotoxic Effect and Molecular Target of Anredera cordifolia

Leaf Extract on HeLa Cervical Cancer Cells

Agustina Setiawati1*, Dita Maria Virginia1, Sri Hartati Yuliani1, Roni Permana Saputra2

1 Faculty of Pharmacy, Sanata Dharma University, Yogyakarta 55281, Indonesia

2 Indonesian Institute of Sciences, Bandung 40135, Indonesia


Discovering selective anticancer agent against cervical cancer from plants

is very challenging. Our previous study reported Anredera cordifolia had

cytotoxic activity against HeLa cervical cancer cells with IC50 75 µg/mL.

However, the selectivity effect and molecular target still remain questioned.

The objective of this study is to determine selectivity as well as molecular

target of Anredera cordifolia leave extract (ALE) on Vero cells, a model of

normal cells. The extract was prepared by macerating Anredera cordifolia

leave powder in 70% ethanol. Viability of cancer cells was measured with

MTT method while cell cycle analysis was calculated with flowcytometry.

Based on the data of cell cycle analysis, molecular target of the extract was

investigated by immunocytochemistry. This study revealed ALE had

selective cytotoxic activity on HeLa cells compared to Vero cells with

Selectivity Index (SI) 17.36. It arrested cell cycle on G1/S phase with Bcl-

2, anti-apoptotic protein, as a molecular target. Therefore, ALE is a

promising to be developed as anticancer agent on cervical cancer.

Keywords:




51

P-02

Genetic Diversity of Damselfly Ischnura aurora Brauer in Ranu Pani

and Ranu Regulo Lake at Semeru Mountain East Java Using

Morphometric Analysis

Andri Maulidi1*, Fitra Arya Dwi Nugraha1, Swiss Winasis1,2, and Nia Kurniawan1

1 Biology Department, Faculty of Mathematics and Natural Sciences, Brawijaya University,

Malang, Indonesia 2 Baluran National Park, Bantangan, Situbondo, East Java, Indonesia


Ranu Pani lake have been destroyed by the human activities, while Ranu

Regulo still in good condition. The difference among both may lead to the

difference of genetic diversity between these areas. This study examine the

genetic diversity of damselfly Ischnura aurora based on morphometric

analysis. Eight morphometric characters were measured. The data was

analized using T-Test, biplot and cluster. Result through T-Test showed

that I. aurora in both areas were not different, but there was a sex

dimorphism between male and female I. aurora. Result through biplot test

showed that distribution of I. aurora was same. Dendogram showed that

most of I. aurora made one cluster. All data shwoed that the diversity of I.

aurora in Ranu Pani and Ranu Regulo lake was low.

Keywords: Genetic diversity, Ischnura aurora, morphometric analysis.



Malang

P-03

Effect of Protein CSN1S2 From Etawah Goat Milk in Diabetic Rat

Model Type 1 Hepar Tissue

Anggi Suci Febriani1 and Fatchiyah1

1 Biology Dept., Math. and Natural Science Faculty,

Brawijaya University


This study aims to analyze how the effects of CSN1S2 protein taken from

goat milk on liver tissues of rat with diabetes mellitus type 1. Animals were

divided into 5 groups divided into control (C), control with CSN1S2 goat's

milk (CM), and diabetes mellitus rat induced by STZ (DM-60 mg/kg),

diabetes given with CSN1S2 goat’s milk protein (DMM-800 mg / kg) and

diabetes given with Glibenclamide (DMG-5 mg / kg). Then, liver’s

histopathology slide was made. Then, stained with HE and visualized using

a microscope. After that cells being calculated. The results are represented

through the histopathology‘s cross-section and the number of normal cells

compared to cells which were damaged. The result shows that the normal

liver tissue has the shape of normal cells with poliferation. While in the

liver tissue of type 1 diabetes, the cells were damaged. The result indicates

that the liver tissue of mice with type 1 diabetes had a histological profile

similar to the liver tissue from control rat. The normal cells proliretation

upregulated and abnormal cells downregulated after goat milk CSN1S2

protein.

Keywords: CSN1S2, diabetes melitus, goat milk, histopatology, liver.




53

P-04

Antioxidant Potential of Red Fruit (Pandanus conoideus Lam.)

Towards Necrosis of Sitotropoblast and Sinsitiotropoblast Cells in

Pregnant Mice (Mus musculus) Before Exposed by Lead

Portia Sumarsono1 and Widjiati1

1 Airlangga University, Surabaya, Indonesia


Lead (Pb) poisoning in animals mainly occur after grazing on contaminated

pasture of lead. Lead was able pass through placenta barrier membrane and

damaging the fetal development which also could induced ROS and

increased apoptosis of placenta. This research aims to determine of

potential of antioxidant from red fruit towards placenta in pregnant mice

before exposed by lead. This research was a laboratory experimental study

using pregnant mice (Mus musculus) as animal research divide by five

treatment groups and four replications each. The negative control group

were not exposed by lead, P0 were exposed by lead 0,011 mg / 20 g BW

each day mice dissolve in 0.1 ml of distilled water and given by oral in

gestation day 6-15. However, the treatment group, pregnant mice were

given red fruit with 0.3 ml/20 g BW, 0.8 ml/20 g BW, 0.9 ml/20 g BW by

oral before exposed by lead. After exposure, the final step were the

preparation of HE staining to calculate the number of remaining normal

sitotropoblast and sinsitiotropoblast cells. The results indicated that the

treatment group were given the antioxidants of red fruit of 0.8 ml / 20 g

BW and 0.9 ml / 20 g BW showed the increase of normal sitotropoblast and

sinsitiotropoblast cells compared with P0 which not given the antioxidants

of red fruit. The antioxidants of red fruit can potential to increase the

number of normal sitotropoblast and sinsitiotropoblast cells in pregnant

mice before exposed by lead.

Keywords: lead, red fruit, sitotropoblast, sinsitiotropoblast



Malang

P-05

The Effect of Water Containing Lead ion (Pb) Against

Malondialdehida (MDA) levels and Duodenum Histopathology In Rats

(Rattus norvegicus)

Reza Albert Patria1, Wibi Riawan2, Dyah Kinasih Wuragil1 and Aulanni’am Aulanni’am1*

1 Faculty of Veterinary Medicine, Brawijaya University 2 Faculty of Medicine Brawijaya University


Lead ion (Pb) can be found on Polyvinyl Chloride (PVC) pipe is a plastic

that usually used for water pipe. It caused toxicity in our body. This study

is aimed to determine the effect of adding water which is containing lead

ion of some Polyvinyl Chloride (PVC) pipes on malondialdehyde (MDA)

levels and duodenal histopathology on rats (Rattus norvegicus). This

research was divided into 5 groups of treatments, were: control, group with

5 mg of Pb/rat/day for 45 days, and third group has given water that already

inside the PVC pipe type A for 60 days, forth group has given water that

already inside the PVC pipe type B for 60 days, and fifth group has given

water that already inside the PVC pipe type C for 60 days. MDA level was

measured by spectrophotometry method and histopathology duodenum

using HE staining and was observed by villi structure, epithelial cells , and

goblet cells. MDA levels were analyzed using One Way ANOVA and

continued with Tukey test (P<0.05). The result of this research indicates

that the use of PVC pipe type A did not significantly affect the increase in

MDA levels and changes in the villi structure, epithelial cell damage and

goblet cell descent in the duodenum histopathology.

Keywords: Duodenum, lead ion (Pb), MDA level, PVC.




55

P-06

Optimization Doses and Time Duration of Oral Glucose in Male

Sprague Dawley Rats

Risqa Novita1 and Sehatman1

1 Centre of Biomedic and Basic Health Technology, National Institute Health Research

Development, Ministry of Health

Jl. Percetakan Negara 23 Jakarta, Indonesia

*Corresponding E-mail: [email protected], risqa @litbang.depkes.go.id

Diabetes Mellitus type 2 case increased fastly in the world, Indonesia had

8.4 million cases on year 2000 and stayed in 4th on the higesht diabetes case

on the world. Based on the fact, we really need optimal preventive, by the

research on animal models. Rats could be infected on DM type 2 by the

giving of glucogenic agents like glucose, but we don’t know yet how long

high titer of blood glucose persists above from the normal titre. This study

used Sprague Dawley rats which is easily find in Indonesia, male, and their

age were 6 weeks with the doses of glucose were 2 gram/bw, 2,5 gram/bw,

3 gram/bw and 3,5 gram/bw consecutive for 7 days. Before they gave the

glucose, they fasted for 12 hours. The measurement of blood glucose did on

0, 30 minutes, 60 minutes, 90 minutes and 120 minutes. After that, we

observed the rats as long as two months for measured their blood glucose

and body weight every weeks. The result of these study showed that the

average of fast blood glucose were above 150 mg/dl but could not persist

longer than 2 hours. The conclusion was, the study did not have good result

for animal models for DM type 2 study yet.

Keywords: Diabetes Mellitus tipe 2, Sprague Dawley rats, Oral Glucose




Malang

P-07

Anti-inflammatory Role of Bioactive Peptide Alpha-S2 Casein Protein

from Goat Ethawah Breed Milk via JAK-STAT3 Signaling

Transduction Cascade

Rista Nikmatu Rohmah1, Ferlany Hardiyanti1 and Fatchiyah Fatchiyah1*

1 Department of Biology, Faculty of Science, Brawijaya University, Jl. Veteran, Malang

65145, East Java, Indonesia


This study aims to analyze the role of STAT3 mechanism and to

design modeling structure the bioactive peptide against STAT3 that

cause inflammation on ileum R A. The ileum was isolated DNA and

amplification with specific primer. The sequence was analyzed using the

Sanger sequencing method. Modeling 3D-structure was predicted by

SWISS-MODEL and virtual interaction was analyzed by docking system

using Pymol and Discovery Studio 4.0 software. This study showed that

STAT3 has target gene 480bp. The normal group and normal treating-

CSN1S2 of goat milk have similarity from gene bank. Whereas, RA group

had transversion mutation that the purine change into pyrimidine even

cause frameshift mutation. Interestingly, after treating with the CSN1S2

protein of goat milk shows reverse to the normal acid sequence group.

Based on in silico study, from eight peptides, only three peptides of

CSN1S2 protein, which carried by PePT1 to enter the small intestine. This

study suggested that the fragment 182-KISQYYQK-189 peptides from

Ethawah goat milk may act as an anti-inflammatory agent via JAK-STAT3

signal transduction cascade at the cellular level.

Keywords: Goat milk Casein, Ileum, Inflammation, Rheumatoid Arthritis,

STAT3.




57

P-08

The Comparative Performance Production of Peranakan Etawah and

Senduro Goats in Controlled Station Farm

Teguh Wicaksono1, Gatot Ciptadi2, Tri Eko Susilorini2 and Abdul Rokhim1

1 The Technical Implementation Unit Services of Livestock Breeding and Animal Feed,

Malang, Indonesia 2 Faculty of Animal Husbandry, UB- Malang, Indonesia 65145.


The local goat of Indonesia namely as Senduro and Peranakan Etawah (PE)

is being good developed in East Java. This research was conducted at the

Technical Implementation Unit Livestock Breeding and Forage Animal

Feed, Malang (UPT-HMT Malang) in order to determine a comparison of

birth weight, body weight gain, and weaning weight and compare the

performance PE and Senduro goats. The material used was a kid PE and

Senduro goat at first parity. Research using the comparative method by

comparing data birth weight, daily weight gain and weaning weight of data

owned and recorded by HMT UPT Malang. The results showed that PE

goat birth weight was greater than Senduro goats are 3.56 + 0.56 kg: 3.25 +

0.66 kg. While body weight gain and weaning weight Senduro was greater

than PE goat respectively - each 0,105 + 0,040 kg / day: 0.122 + 0.055 kg /

day and 13.00 + 3.87 kg: 14.31 + 5.19 kg.

Keywords: Local Goat, Indonesia, growth. birth weight, weaning weight.



Malang

P-09

Profile of a Blastocyst Protein: Pregnancy-Associated Glycoprotein

(PAG)

Tita Damayanti Lestari1*

1 Department of Veterinary Reproduction, Faculty of Veterinary Medicine

Universitas Airlangga, Kampus C Unair Jl. Mulyorejo Surabaya


Protein blastocyst named pregnancy-associated glycoprotein (PAG) has

been isolated from pregnant dairy cows serum. Isolation was conducted to

find out the protein characterization. Protein PAG were isolated from six

pregnant cows serum of 1 – 5 days prior to give birth. Characterization of

PAG was done by SDS -PAGE technique. Gel electrophoresis from SDS -

PAGE revealed some bands which were recognized by Standard Marker of

Anti-PAG. Determination of molecular weight was based on Retardation

factor (Rf) calculation and was done by Western Blot method. Level of

PAG protein was determined by Biuret method. Protein contained of PAG

was defined through ELISA analysis. Result shows PAG has molecular

weight of 67.34 kDa, contains protein of 4740 g/ml and PAG level is

2412 g/ml.

Keywords: Anti -PAG Standard, Pregnant cows serum, PAG, SDS-PAGE,

Western Blot.




59

P-10

Darapladib Administration Effect of Perivascular Adipose Tissue

Thickness in Type 2 Diabetes Mellitus Model Sprague-Dawley Rats

Titin Andri Wihastuti1*, Teuku Heriansyah1, Kenty Wantri Anita1 and Febriana R1

1Medical Programs, Faculty of Medicine, Brawijaya University, Malang, East Java, 65145,

Indonesia


Cardiovascular disease (CVD) take the first place for causing death

worldwide with 29% from all causes of death. CVD often caused by

atherosclerosis plaque within vascular. Insulin resistance is a risk factor risk

in atherosclerosis occurrence which is available in type 2 diabetes mellitus

condition. As for the causative roles of atherosclerosis are complex because

it is affected by many factors. One of the causative is Lp-PLA2, a novel

marker for atherosclerosis progressivity. Lp-PLA2 enzyme hidrolyzes LDL

to become ox-LDL and ox-NEFA which then will attract macrophages and

other pro-inflammatory substances into lesion. So, the inflammation will

rise and trigger endothel dysfunction. The dysfunction of endothel will

induce the production of perivascular adipose tissue (PVAT) so it will

thicker. Until now, it has been found one of the novel therapies that inhibits

Lp-PLA2 with Darapladib oral drug. This study used post-test only

controlled group design. 30 Sprague Dawley rats were divided into three

groups which was normal group, type 2 DM model group, and type 2 DM

model with Darapladib administration group. Each group was divided into

two serial time which was 8 weeks (early phase) and 16 weeks (late phase).

The Parameters in this study is PVAT thickness. There were shown an

insignificant role (p > 0,05) of Darapladib with PVAT thickness (repeated

ANOVA). But there was also shown a decreasing pattern between the

groups. According the result, it is suggested to add one more group which

is the 0-week group for increasing the data’s reliability.

Keywords: Darapladib, Perivascular Adipose Tissue, Lp-PLA2,

Atherosclerosis, Type 2 Diabetes Mellitus.



Malang

P-11

Effectiveness of Giving Hematopoietic Stem Cell in Pregnant Mice Exposed by

Lead towards Embryo Resorption and Defects Conginental

Widjiati1

1Airlangga University, Surabaya, Indonesia


The purpose of this research was to prove the effectiveness of Stem Cell

Hemopoetic in pregnant mice that expose by heavy metals as a model

teratogenic to reducing embryo resorption and defects conginental. This

research object was pregnant mice that exposed lead acetate and treated

with Stem Cell Hemopoetic, further examination, include: examination of

the number of embryonic resorption and defects conginental. This study

was divided into 2 groups: treatment group (P1) pregnant mice were given

eight days to MEM medium and given intravenous doses of 1/16 LD50

lead during gestation days 9-15, P2 pregnant mice were given a day to 8

Hemopoetic Stem Cell 1x107 and given intravenous doses of 1/16 LD50

lead during gestation days 9-15. The results showed that Stem Cell

Hemopoetic can reduce the number of defective embryo and conginental. It

is concluded that the provision of Stem Cell Hemopoetic on pregnant mice

exposed by lead may decrease resorption of embryo and disability

conginental.

Keywords: Stem Cell Hemopoetic, lead, teratogenic, embryo resorption,

defect conginental.




61

COMMITTEE

3rd

International Conference of Advance Molekular Bioscience & Biomedical

Engineering (3rd

ICAMBBE 2016)

August, 4th

-5th

2016

Steering Committee

• Rektor

• Pembantu Rektor I

• Pembantu Rektor II

• Prof. Fatchiyah, M.Kes., Ph.D.

• Prof. Dr. drh. Auliani’am, DES

• Gugus Irianto, Ph.D., AK

Organizing Committee

Head of Committee • Dyah Kinasih Wurangil, MP., M.Sc.

Treasurer • Dr. Ir. Gatot Ciptadi, DESS

• Perdana Finawati, S.Si

• Ninik Afriza, M.Si

• Sri Tanjung Bilgis

Secretariat &

Promotion

• drh. Herlina Pratiwi, M.Si

• Lidwina Faraline T., M.Si

• Megawati Sistin Agustita, S.Si

• drh. Fitria Novitasari

Scientific

• Irfan Mutofa, M.Si., Ph.D.

• drh. Dyah Ayu Oktavianie, M.Biotech

• Pupimadita Tizar Afdora, M.Si

Accommodation&

Hospitality

• Anita Herawati, M.Si

• Wahyu Nur Laili Fajri, M.Si

• Rista Nikmatu Rohmah, M.Si

• Ninuk Mahmudah

Publication &

Documentation

• Yulianto M. Nugroho, S.Si

• Akbar Farid H., S.Si

• Matsyinatul Khoiriyah S. S

• Oky Dian, SIP

Transportation

• Bayu

• Zainul Hadi

• Abdul Jaelani



Malang

Accessibility

Public Transportation UB Guest House

Kota Baru Station – ADL (blue color) – UB (Mayjend Panjaitan Street)

Kota Baru Stasion – AL (blue color) – UB (Veteran Street)

Bus Track

Arjosari Bus Station – ADL (blue color) – UB (Mayjend Panjaitan Street)

Arjosari Bus Station – AL (blue color) – UB (Veteran Street)

Landungsari Bus Station – ADL/LDG (blue color) - UB (Mayjend Panjaitan

Street)

Landungsari Bus Station – AL/GL (blue color) - UB (Veteran Street)

Taxi

Citra Taxi : 0341 – 490555

Argo Taxi : 0341 – 488888

Mandala Taxi : 0341 – 474747

Bima Taxi : 0341 – 364026

Contact Person

UB Guest House

Jl. M.T. Haryono No. 169, Ketawanggede, Kec. Lowokwaru, Kota Malang

East Java – Indonesia

Phone: 0341 558585




63

Site Map

General Facilities of Guest House UB

Keterangan:

UB Hotel Politeknik Negeri Malang

Universitas Brawijaya Malang Taman Krida Budaya

Malang Town Square



Malang

Supported by:

Contact Info:

PT BANK MANDIRI (PERSERO) Tbk. Jl. Jenderal Gatot Subroto Kav. 36-38

Jakarta 12190 Indonesia

Telp: 14000, +62-21-52997777

Fax: +62-21-52997735

Contact Info:

CV Gamma Scientific Biolab

Jl. Bandara Abd Saleh GF No. 29

Malang 65138, Jawa Timur

Indonesia

Contact Info:

MAKMUR SEJATI

Perum Griya Shanta Blok L 238

Malang

Contact Info:

PT. UNILEVER INDONESIA, Tbk.

Jl. Jend. Gatot Subroto, Kav. 15,

Jakarta 12930, Indonesia

Contact Info:

PT. DEMKA SAKTI

Jl Tanah Abang III No. 19 A-B

Jakarta Pusat




rd international conference on advance molecular...

Documents