Project proposal using FETAX to examine the buffering qualities of docosahexaenoic acid (DHA) against acrylamide (ACR) exposure. (this presentation does not include data)

1. Using FETAX to examine the buffering qualities of
docosahexaenoic acid (DHA) against acrylamide
(ACR) exposure.
VANESSA CHAPPELL

2. • It is currently classified as a Group 2A, probable
human carcinogen by the EPA. However, it is a
known teratogen and neurotoxin and a potential
genotoxin
• It can be formed when amino acids interact with
sugars during cooking (bread, cookies, cereal,
etc.)
• It can also be formed when fats are oxidized
during deep frying.
• Repeated or prolonged exposure can produce
target organ damage
What is ACR?

3. Where is it found?
Industry
● Electrophoresis gels
● Paper and plastic products
● As binders or thickening agent
● Polymer manufacturing
● Cosmetics and lotions
● Waste water treatment
● Pesticide formulation
● Dyes and adhesives
● Cigarettes (including secondhand)
Food
● Coffee
● Bread
● Potato chips and fries
● Food made from grains
● Cookies
● Processed foods
● Sugar manufacturing
● Water

4. So what can we do to protect
ourselves from ACR exposure?
Diet is our biggest environmental exposure so
why not start there? After all we can't quit eating
and breathing, right?
Hypothesis:
The effects of ACR exposure can be buffered by
supplementing the diet with DHA

5. What is DHA?
• Poly unsaturated omega –3 fatty acid
• It is the primary structural component of the
brain, cerebral cortex, skin, sperm, testicles
and retina.
• Improves egg quality and It is required for
embryo development
• DHA deficiency is associated with cognitive
decline and increased neural cell death.

6. • Americans get less than 50%
RDA of Omega-3
• Ancestors ratio of Omega-3's
to Omega-6 was 1:1
• This ration is now 1:10
Why is this important?

7. Why use a frog?
Life Cycle
Early development has close
evolutionary relationship to humans
96 hour test
FETAX testing is inexpensive and
fast
Harvesting Eggs
Each egg harvest produces
hundreds of subjects. Each pair can
be bred 3-4 times/year

8. Xenopus laevis
The African Clawed Foot Toad

9. What is FETAX?
(Frog Embryo Teratogen Assay-Xenopus)
Developmental
Toxicology Test
• Malformation
• Morbidity
Solution Concentrations
Increasing concentrations of a
chemical are used to observe
abnormalities
FETAX solution
• NaCl
• NaHCO3
• KCl
• CaCl3
• CaSO4+2H2o
• MgSO4

10. FETAX
Frog Embryo Teratogen Assay on Xenopus –a 96 hour test to observe malformation and mortality of embryos. The embryos are
exposed to increasing concentrations of a chemical to observe if any abnormalities occur.

11. Egg Collection
● Egg production is manipulated by
hormone injections approximately 12-
16 hours before desired FETAX
testing time
• Eggs are dejellied
using Cysteine at a
pH8

12. Test Maintenance
Solutions
New solutions are made daily and
each dish is changed
Mortalities
Daily records of death
Malformations
At 96 hours any malformations
were recorded
20 eggs are placed in each petri dish

13. Analysis
MS222
Used to anesthesize the tadpoles
in order to obtain pictures so that
measurements can be made.
Image Pro Plus
Utilized to perform length
measurements
SYSTAT
Used to analyze data

14. Analysis
Further analysis is done using AVONA
with Bonferoni test and Probit analysis
LC50, EC50 and No observable effects
data are determined

15. Future Research
• Biospecimen collection for human epidemiological cohort studies to determine
ACR and PUOFA levels before adding DHA supplement, then after 6-8 weeks and
after 12-16 weeks of DHA supplementation
• iron deficient patients and PUOFA levels
• Possible phytochemical studies to determine synergistic benefits with DHA