effects of bromelain in pineapple juice - SGU Repository

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By

Anjelica Marsha Tee 14310054

A thesis submitted to the Faculty of LIFE SCIENCES AND TECHNOLOGY

In Partial Fulfillment of the Requirements for the

BACHELOR’S DEGREE in

BIOMEDICAL ENGINEERING

SWISS GERMAN UNIVERSITY EduTown BSD City

Tangerang 15339 Indonesia

August 2014

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STATEMENT BY THE AUTHOR

I hereby declare that this submission is my own work and to the best of my knowledge, it contains no material previously published or written by another person, nor material which to a substantial extent has been accepted for the award of any other degree or diploma at any educational institution, except where due acknowledgement is made in the thesis.

Anjelica Marsha Tee

_____________________________________________

Student Date

Approved by:

Dr. rer. nat. Maruli Pandjaitan

_____________________________________________

Thesis Advisor Date

Dr. Janet Julianita, Sp. PA

_____________________________________________

Thesis Co-Advisor Date

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ABSTRACT

EFFECTS OF BROMELAIN IN PINEAPPLE JUICE TO ABNORMAL CELLS IN CERVIX

By

Anjelica Marsha Tee

Dr. rer. nat. Maruli Pandjaitan, Advisor Dr. Janet Julianita, Sp. PA , Co-Advisor

SWISS GERMAN UNIVERSITY

Cell inflammation can be causes several factors. In the cervix, the cell inflammation may causes by virus and bacteria. This research was focused to see the effect of enzyme Bromelain in pineapple juice to abnormal cells. Based on previous research, enzyme Bromelain can break down the capsid of HPV.

The methodology that was used in this research was Pap smear, HPV DNA test, Polymerase Chain Reaction. From 400 patients in 4 public health centres or Puskesmas in Bogor, 24 patients were suspected to cervix inflammation.

Based on the result of Pap smear test, 4 patients were found to have cell inflammation. Then all four patients were treated with daily consumption of pineapple juice. DNA test of HPV were tested to check whether the inflammation was caused by HPV. From the microscopic image, it can be seen that there was minor changes in the inflammation cells and the cervix cells.

Keywords: Cervix, Cell inflammation, Bromelain, Ananas comosus, Human Papilloma Virus, Bacteria

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DEDICATION

I dedicate this to everyone who has lost a daughter, a mother, a grandmother that is suffered from cervix cancer

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ACKNOWLEDGEMENTS

First of all, I would like to give thanks to the Almighty God for giving me strength and guidance to overcome all obstacles and difficulties during the making of this thesis.

Secondly, I would like to give thanks to Dr. rer. nat. Maruli Pandjaitan for being my Advisor, for being patient and giving me support, guidance and answers to all obstacles.

I would also like to thank my Co-Advisor, Dr. Janet Julianita Sp. PA for her insight in pathological knowledge and her time to examine the samples.

I would like to thank the Dean of Life Sciences, Mr. Irvan Kartawiria ST. M.Sc, for his support to finish this thesis.

I also want to say thanks to all staffs in Dinas Kesehatan Bogor, especially Drg.

Lindawati MKM, Mrs. Pinna Situmorang, for their help me to distribute the pineapple juice.

I want to express my gratitude towards Mr. Tabligh Permana, M.Si for the guidance, patience and help to answer all questions during the experiments.

I thank my family for their endless patience, help and support during this time.

Lastly, I would like to thank the entire Life Sciences batch 2010, especially to Melliany, Tika, Nabila, Irene, Jason, Adit, Gerald, Raka, Sandy, Andika, Andre, Joeyanti and Geoffrey that always listen to my problems and giving me laughter.

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Page

STATEMENT BY THE AUTHOR ... 2

ABSTRACT ... 3

DEDICATION ... 5

ACKNOWLEDGEMENTS ... 6

TABLE OF CONTENTS ... 7

LIST OF FIGURES ... 9

LIST OF TABLES ...10

CHAPTER 1 – INTRODUCTION ...11

1.1.Background... 11

1.2.Research Problems ... 12

1.3.Research Objectives ... 12

1.4.Significance of Study... 13

1.5.Research Questions ... 13

1.6.Hypothesis ... 13

CHAPTER 2 – LITERATURE REVIEW ...14

2.1. Anatomy of Cervix ... 14

2.2. Cells Inflammation ... 15

2.2.1. Bacteria ... 17

2.2.2. Human Papilloma Virus ... 18

2.3. HPV Detection ... 19

2.3.1. Pap Smear ... 19

2.3.2. HPV DNA Test ... 21

2.3.3. Gel Electrophoresis ... 22

2.4. Pineapple Fruit ... 23

2.5. Enzyme Bromelain ... 24

CHAPTER 3 - RESEARCH METHODS ...26

3.1. Venue and Time ... 26

3.2. Materials and Equipment ... 26

3.2.1. Pap smear and HPV DNA sample collection ... 26

3.2.2. Pap smear staining... 26

3.2.3. HPV DNA detection ... 27

3.2.4. Determining Enzyme Bromelain Protein Content ... 27

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3.3. Overall Procedure ... 28

3.4. Preliminary Research ... 29

3.4.1. Patients Data Collections ... 29

3.4.2. Preparation of Pineapple Juice ... 29

3.5. Experimental Procedure ... 30

3.5.1. Sample Collection... 30

3.5.2. Pap smear Staining ... 31

3.5.3. Data Analysis ... 32

3.5.5. Amplification of sample by using Polymerase Reaction Chain ... 33

3.5.6. Gel Electrophoresis and Documentation ... 34

3.5.7. Protein Content of Enzyme Bromelain on Pineapple Juice ... 35

3.5.8. Volume Activity of Enzyme Bromelain on Pineapple Juice ... 36

3.6. Observations ... 37

3.7. Analytical Procedure ... 37

CHAPTER 4 – RESULTS AND DISCUSSION ...39

4.1. Patient Examination Results ... 39

4.1.1. Before Treatment ... 39

4.1.2. After Treatment ... 41

4.2. Specific Activity of the enzyme Bromelain ... 45

4.2.1. Volume Activity ... 45

4.2.2. Protein Content ... 46

4.2.3. Specific Activity ... 46

CHAPTER 5 – CONCLUSIONS AND RECCOMENDATIONS ...48

5.1.Conclusions ... 48

5.2.Recommendations ... 48

GLOSSARY ...49

REFERENCES...51

APPENDICES ...53

APPENDIX 1 ... 53

APPENDIX 2 ... 55

APPENDIX 3 ... 56

CURRICULUM VITAE ...58

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Figures Page

2.1 Anatomy of female reproduction system (Terese Winslow, 2009) ...14

2.2 Cervical transformation zone (WebMD, 2012) ...15

2.3 Three dimensional model of HPV (Henderson, 2001). ...18

2.4 Human Papilloma Virus life cycle (The National Institute for Medical Research, 2004) ...19

2.5 Principles of Polymerase Chain Reaction... 22

2.6 Pineapple fruit ... 24

3.1 Flowchart of overall procedure from the experiment ... 28

4.1 PCR result of suspected HPV positive samples ... 40

4.2 The visualization PCR of suspected samples by Natalia Budihardjo in 2010 ... 40

4.3 Pap smear microscopic image of patient no. 3 ... 42

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LIST OF TABLES

Table Page

2.1 Besthesda System of Cytological Classification (Besthesda System

Terminology, 2011) ... 20

3.1. PCR mixture ingredients ... 33

3.2. PCR cycle procedure description ... 34

3.3. Variation Concentration of Bovine Serum Albumin ... 35

3.4. Overall Steps of Volume Activity Test ... 36

4.1. Initial Patient Examination ... 39

4.2. Patient Examination Result after Treatment ... 41

4.3. Absorbance and Volume Activity of Enzyme Bromelain in Pineapple Juice ... 45

4.4. Absorbance and Protein Content of Enzyme Bromelain in Pineapple Juice ... 46

4.5. Specific Activity of Enzyme Bromelain in Pineapple Juice ... 47