Capstone
5/8/15
Methods: Detailed Outline
Fucoidan consumption associated with slowing the progression of breast cancer
I.
Participants
A.
Clinical single blinded study was approved to study the effects of seaweed
consumption on breast cancer patients
i.
The Human Subjects Research and Review Board of West Coast
University and Material Command (Los Angeles, California)
ii.
The Jackson Memorial Hospital (Miami, Florida)
iii.
West Coast University Institutional Review Boards
iv.
IRB number, 91-027
v.
Clinical registration number WCU4166852
B.
18 diagnosed female breast cancer patients recently diagnosed in stages 3 and 4
referred to our clinical trial
i.
9/18 had stage 3 breast cancer
ii.
9/18 had stage 4 breast cancer
II.
C.
Control group consisted of 10 female breast cancer patients in any stage of cancer
D.
Inclusion Criteria
i.
Breast cancer patients between the ages of 30-70 years old
ii.
Participants must have no allergies to seaweed, shellfish, iodine, or soy
iii.
Participants must not have been on hormone replacement therapy,
radiation therapy, or chemotherapy within the previous six months of the
study and no history of cancer (other than breast cancer) within the past
twenty years
iv.
Participants must not have diabetes or gastrointestinal disorders
v.
Alcohol intake limited to less than one per week
vi.
No oral antibiotics taken within the last three months
vii.
Women agree to consume their normal diet, avoiding seaweeds and to
continue their habitual use of medications, chemotherapy, vitamins and
supplements during the study
Description of breast cancer stage under investigation
A.
Stage 3 Breast Cancer
i.
Cancer extends beyond immediate region of the tumor
ii.
Invades nearby lymph nodes and muscles but has not spread to vital
organs
iii.
Divided into three groups based on the size of the tumor: Stage 3A, 3B,
and 3C
iv.
Stage 3A falls under three distinct descriptions
1
v.
vi.
B.
III.
a. Tumor is less than 2cm and has spread to 4-9 lymph nodes
b. Tumor is larger than 5cm and cancer clusters found in lymph nodes
c. Tumor is larger than 5cm and has spread to lymph nodes near the
breastbone or underarm
Stage 3B description
a. Tumor size is variable and can be any size
b. Cancer has spread to the chest wall or breast skin
c. Cancer has spread to lymph nodes
d. Evidence of swelling, inflammation or ulcers is visible
e. Cancer has invaded at least 9 lymph nodes
Stage 3C description
a. No tumor or a tumor of variable size and cancer has invaded at least 10
or more lymph nodes
b. No tumor or a tumor of variable size and cancer has spread to lymph
nodes near collar bone
c. No tumor or tumor of variable size and cancer has spread to lymph
nodes near underarm or breastbone
Stage 4
i.
Cancer spread to other parts of the body, such as the brain, bones, lung
and liver
ii.
This stage is considered incurable
Study Design
A.
Clinical study spanned a total of 5 years from April 2015-April 2020
B.
The ten control group breast cancer participants ingested a sugar placebo pill as
their medication
C.
The remaining 18 breast cancer patients ingested 5 grams per day for a total of 10
500 mg capsules
i.
Seaweed capsules were obtained from a manufacturer based in Okinawa, Japan
IV.
Health and Mortality Surveillance
A.
Pre-evaluation tests were taken prior to the start of treatment to establish each
participant’s baseline values
i.
Colony formation, MTT assay and xenograft to determine breast cancer cell
numbers (EXPLAIN)
ii.
Blood tests- prior confirmatory results that diagnosed breast cancer patients
a. Hormone receptor test- tests if the breast cancer cells have receptors for the
hormone estrogen and progesterone
b. HER2/neu test (ERBB2 test)- known as the human epidermal growth factor
receptor 2 proto-oncogene. This is usually overexpressed and amplified in
invasive breast tumor tissue
2
iii.
Mammogram
a. X-ray picture of the breast
b. Used to check breast cancer who show no signs or symptoms of the disease
c. Used to check for breast cancer progression
d. Description of Procedure- breasts are compressed between two firm surfaces
to spread out the breast tissue. The x-ray takes black and white images of the
breast
B.
Daily seaweed capsule ingestion at 5 grams per day (ten 500 mg capsules per day)
C.
Every 6 months post treatment, colony formation, MTT assay, xenograft and cell
number evaluation was done to assess improvement or no improvement in breast cancer
tissue and cells
i.
Explain colony formation
ii.
Explain MTT assay
iii.
Above tests will determine cell numbers of breast tissue seen
D.
Every 3 months/90 days, CT scans were done to check for tumor growth
i. Explain CT scan
E.
Monthly journal log that documented food intake, exercise/fitness routines and
mood. This journal was given to case manager for review and discussion at monthly
check up.
F.
Monthly support and rehab meeting to offer program support to breast cancer
survivors
G.
Every four months breast cancer patients had a mammogram done to detect the
presence or absence of cancer cell growth
V.
H.
In the case that participants passed away, their death certificates were requested
I.
After five-year study, post evaluation tests were done to include colony formation
assay and MTT assay on 24 and 48 hours to determine cell number differentiation
Data Analysis
A.
Calculation of Participant Survival Rates
i.
2/10 participants dropped out who were part of the placebo group due to
non-compliance
ii.
23 total survivors remained
iii.
3/18 participants died. 2 died who was diagnosed with stage 4 cancer and
1 died who was diagnosed with stage 3 cancer
B.
Calculation of Participant Retention Rates
i. 23/28 participants remained in the 5-year clinical study
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C.
Tumor Grading Description
i.
Use of Nottingham grading system - this grading system is a reference tool
to describe tumor biology
ii.
Low-grade tumors (DESCRIBE)
iii.
High-grade tumors (DESCRIBE)
D.
Tumor Size Evaluation
i.
Comparison of 4T1 Wnt/B-Catenin (estrogen receptor positive), MDAMB 231 Wnt/B-Catenin (estrogen receptor positive), and MCF7 activation
of capsase-8 (estrogen receptor positive signaling pathway) cancer cell
lines to monitor the proliferation, cell differentiation or apoptosis of breast
cancer cells
ii.
CT scan (CAT scan, also known as computerized tomography scan) to
check growth of tumor
a. x-ray technique to provide health care providers 2-dimensional cross
sections of internal body parts
iii.
In Situ TUNEL Assay to check if fucoidan induced apoptosis in tumor
tissue
a. TUNEL assay tests DNA fragmentation by light microscopy
REFERENCES
About Breast Cancer. (n.d.). Retrieved May 5, 2015, from
http://www.nationalbreastcancer.org/about-breast-cancer
4
Funahashi, H., Imai, T., Mase, T., Sekiya, M., Yokoi, K., Hayashi, H., Shibata, A., Hayashi, T.,
Nishikawa, M., Suda, N., Hibi, Y., Mizuno, Y., Tsukamura, K., Hayakawa, A. and
Tanuma, S. (2001), Seaweed Prevents Breast Cancer? Japanese Journal of Cancer
Research, 92: 483–487. doi: 10.1111/j.1349-7006.2001.tb01119.x
Hsu, H., Lin, T., Hwang, P., Tseng, L. Chen, R., Tsao, S., & Hsu, J. (2013). Fucoidan induces
changes in the epithelial to mesenchymal transition and decreases metastasis by
enhancing ubiquitin-dependent TGFB receptor degradation in breast cancer.
Carcinogenesis, 34(4), 874-884.
Sekiya, M., Funahashi, H., Tsukamura, K., Imai, T., Hayakawa, A., Kiuchi, T., & Nakao,
A. (2005). Intracellular signaling in the induction of apoptosis in a human breast cancer
cell line by water extract of Mekau. International Journal Of Clinical Oncology, 10(2),
122-126. doi:10.1007/s10147-004-0469-2
Smyth, P.A. (2003). Role of iodine in antioxidant defense in thyroid and breast disease.
Biofactors, 19(3/4), 121-130.
Teas, J., Vena, S., Cone, D., & Irhimeh, M. (2013). The consumption of seaweed as a
protective factor in the etiology of breast cancer: Proof of principle. Journal of
Applied Phycology, 25(1),771-779. doi: 10.1007/s10811-012-9931-0
Vogel, V.G. (2015). Ongoing data from the breast cancer prevention trials: opportunity for
breast cancer risk reduction. BMC Medicine, 13(1), 1-4. doi:10.1186/s12916-015-0300-0
Yang, Y., Nam, S., Kong, G., & Kim, M. (2010). A case–control study on seaweed
consumption and the risk of breast cancer. British Journal of Nutrition, 103(9), 13451353. doi:10.1017/s0007114509993242
5
Capstone Methods Outline
•
Method
A. Hook: In our stem cell transplant for SCD, stem cells are taken from the patient in utero,
specialized to repair the defective gene, and implanted back into the fetus’s bone marrow.
•
#1 Subtopic: Overview of autologous stem cell transplant
1. Population criteria
2. Risks and benefits of autologous stem cell transplant
3. Surgical methods and tools of normal stem cell recovery and transplant
•
#2 Subtopic: Overview of pinpoint gene therapy
1. Zinc finger nucleases method
2. The study seeks to amend the change in the beta-globin chain so bone marrow stem
cells produce ordinary, circular shaped blood cells
3. Utilized extraordinarily engineered enzymes
•
#3 Subtopic: Overview of fetal surgery
1. Risks associated with surgery
2. Minimally invasive fetal surgery that will be used in experiment
3.
An ultrasound to see the baby movement
4. To inject the baby with zinc fingers to prevent sickle cell
5. Safety and complications.
6. Sign consent for procedure
Capstone Discussion Outline
Sickle cell disease is a hereditary blood disorder that is moved down through families. Both
parents need to carry the sickle cell gene for their child to have it. In sickle cell disease, glutamic
acid in beta chain of hemoglobin is supplanted by valine at position 6. It can bring about sickling
of red blood cells, which lessens their flexibility and oxygen carrying capacity. Typical red blood
cells are round or biconcave disc shaped and highly elastic. There is no generally accessible cure
for Sickle Cell Disease. However, a little number of kids with Sickle Cell Disease have been
cured through stem cell transplant.
Stem cells are cells that can form into a wide range of cells in the body. They serve as a repair
framework for the body. Stem cells are found in bone marrow. This is the elastic tissue inside a
few bones, similar to your hip and thigh bones. Stem cells additionally are found in umbilical
cord blood, the blood in the umbilical cord and placenta. This blood can be collected after the
umbilical cord is cut during childbirth.
In a stem cell transplant for SCD, stem cells taken from a healthy individual are put into a man
with SCD. This system is extremely hazardous and can have genuine reactions, including death.
The stem cell gene therapy system portrayed in the study seeks to amend the change in the betaglobin chain so bone marrow stem cells produce ordinary, circular shaped blood cells. The
method utilized extraordinarily engineered enzymes, called zinc-finger nucleases, to wipe out the
changed hereditary code and replace it with a corrected version that repairs the beta-globin
mutation.
The most recent gene correction method utilized by the group utilizes exceptional proteins,
called zinc-finger nucleases, to truly cut out and remove the hurtful mutation, replacing it with an
amended variant. So after gathered bone marrow stem cells from people with sickle cell disorder.
These bone marrow stem cells would ordinarily give rse to sickle-shaped red blood cells. In any
case, in this study, the group destroyed them with the zinc-finger nucleases keeping in mind the
end goal to redress the mutation.
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