1. Completed PhD Project
Heshu Sulaiman Rahman

2. Parts of the Project Title. Supervisory Committee. Field of the Study.
Location of the Project.
Principles of the Work.
Main Objective.
Chapters and Experiments.
Type of the Thesis.
Examination Committee.
Grants Used.
Result and Achievements.
Special Dates and Courses Involved.

3. Title of the Project
ANTI-LEUKEMIC EFFECTS OF ZERUMBONE NANOPARTICLE ON HUMAN JURKAT T LYMPHOBLASTOID CELL LINE IN VITRO AND MURINE LEUKEMIC WEHI-3B MODEL IN VIVO

4. Supervisory Committee
1. Professor Dr. Rasedee Abdullah (Main supervisor)
Specialist: Haematology and Clinical Pathology
2. Associate Professor Dr. Ahmad Bustamam Abdul (1st Co-supervisor)
Specialist: Pharmacology
3. Associate Professor Dr. Zeenathul Nazariah Allaudin (2nd Co-supervisor)
Specialist: Molecular Virology

5. Field of the Study
Haematology: Haemogram, Bone Marrow and Blood Disorders Interpretation.
Clinical Pathology: Blood Biochemistry Including Liver and Kidney Function Tests, Toxicity Diagnosis, Urinalysis and Cytology.

6. Locations of the Project
University Putra Malaysia (UPM), Serdang, Selangor, Malaysia.
Cancer Research Laboratory (MAKNA), Unit Microscopy, and Immunotherapeutics and Vaccines laboratory (LIVES) (Institute of Bioscience (IBS), UPM): 80%
Haematology Laboratory and Histopathology Laboratory (Faculty of Veterinary Medicine, UPM): 20%

7. Principles of the Work Natural Product for Cancer Therapy.
Nanotechnology for Cancer Therapy (Drug Design, Development and Delivery).

8. Main Objective of the Study
Preparation, characterisation of Zerumbone Nanostructured Lipid Carrier (ZER-NLC) and evaluation its in vitro and in vivo anti-leukemic activities.

10. Chapters and Experiments
Isolation, Purification of ZER and Antileukemia Study:
Isolation (Hydrodistillation method).
Purification (N-hexene).
Purity determination (RF-HPLC)
Cytotoxicity (MTT) of ZER and Doxorubicin on Human lymphoblastoid Jurkat T cell line.
Morphological detection (NIM, SEM, FM).
Proteomic detection (Caspase-3, -8 and -9 activities).
Apoptosis detection using flowcytometry (Annexin V, cell cycle and TUNEL).
Total no. of Experiments: 13

12. 2. Preparation and Characterization of ZER-NLC:
Preparation of ZER-NLC (high pressure piston gap homogenization technique).
Characterization of ZER-NLC:
Particle size, PDI, ZP (Zetasizer).
Particle shape (TEM).
Crystallinity and polymorphism (DSC).
Radiation scattering (Wide angle X-ray diffraction).
Stability, pH, Sterilization,
EE and DLC (HPLC).
In vitro drug releasing study (FDCS).
Total no. of Experiments: 13.

14. 3. In vitro Antileukemic Effects of ZER-NLC:
Cytotoxicity (MTT) of ZER-NLC and Doxorubicin on Human lymphoblastoid Jurkat T cell line.
Morphological detection (SEM, SEM, FM).
Proteomic detection (Caspase-3, -8 and -9 activities).
Apoptosis detection using flowcytometry (Annexin V, cell cycle and TUNEL).
Total no. of Experiments: 10

16. 4. Acute Toxicity Study of ZER-NLC in BALB/c Mice Model
(48 male of 8 groups and 48 females of 8 groups)
Clinical observation and body weight.
Histopathology study (6 organs).
Serum Biochemistry (9 tests for liver function and 5 tests for kidney function).
Hemogram (4 tests).
Bone marrow examination.
Total no. of Experiments: 6 (18 tests)

18. 5. In vitro Antileukemic Effects of ZER-NLC in WEHI-3B Cell Line
Cytotoxicity (MTT) of ZER-NLC and ATA on myelocytic murine WEHI-3B cell line.
Morphological detection (Hoechst 33342).
Proteomic detection (Caspase-3, -8 and -9 activities).
Apoptosis detection using flowcytometry (Annexin V and cell cycle).
Total no. of Experiments: 7

19. 6. In vivo Antileukemic Effects of ZER-NLC on Mice Model
(60 male of 6 groups)
Hemogram (DCC, Hb, PCV, WBC, RBC).
Histopathology for spleen and liver.
TEM for Spleen and liver.
TUNEL for spleen .
Proteomic and genomic detection of blood biomarkers (7 genes).
Total no. of Experiments: 6
Total experiments for 4 and 5: 13

21. 7. Immunological Evaluation of ZER-NLC in Mice Model
(60 male of 6 groups)
Immunophenotyping (NFKB, CD3, CD4, CD8).
Immunohistochemistry (CD3 and CD19).
Immunoflurescence (CD3 and CD19)
d. Oxidative enzymes (SOD, GSH, FARP, iNOS, MDA).
Total no. of Experiments: 6 (13 tests)
Total no. of Experiments in the Project : 61 (excluding no. of tests)

23. Grants Used for the Project

24. Type of Study
By Research
Type (Written Style) of the Thesis
Chapters (6)

25. Examination Committee
Prof. Dr. Rani Bahaman (Chairman).
Prof. Dr. Tenguk Azmi Ibrahim (Internal Examiner).
Prof. Dr. Azmi Noor Laila (Internal Examiner).
Prof. Dr. Fang-Rong Chang (External Examiner).

27. Results Minor Modifications CGPA: 3.965/4
Best PhD Thesis at the Level of UPM for Year (Gold Medal + RM 3000).

28. Achievements Related to this Project
Published papers in Q1 Journals: 6
Published Conference Proceedings: 7
International and National Conferences: 23
Patents: 3
Medals: 3 (2 Gold + 1 Bronze)

30. Courses Involved Advanced Veterinary Haematology.
Special Topics in Veterinary Clinical Pathology.
Animal Molecular Biology.
Animal Patho-physiology.
Ultrastructure and Electron Microscopy
Total no. of Credits: 15

31. Special Dates 1st day of (enrolment) Registration 8.2.2010
1st day of Lab Work
Thesis Submission
Viva Voce
Senate Approval

32. Final Speech for Success
Always be kind.
Do not wait for reward or benefit.
Be hopeful and try your best.

33. THANK YOU