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Wanda figueroa 29nov2011
1. Research Experience:
“Scavenging and Antioxidant Activity Towards
Superoxide Radical of Plant Extracts used as
Diabetes Adjuvant”
University of Puerto Rico at Cayey
Cayey Puerto Rico 00736
Wanda M. Figueroa-Cuilan
4. To investigate the in vitro biological activities
and in vivo efficacy of herbal formulations used as
diabetes adjuvants in Puerto Rico to develop new
therapeutic methods to treat diabetes.
Program Goal
Researches Around the world
Around the world several studies have been
conducted to discover good antioxidant and scavenging
plants extracts, in order to develop new therapeutics to
treat diabetes.
Example: Triphala (2009)
5. Plant Extract IC50 (ug/mL) Country Research Title
Spondias pinnata 13.46 India Antioxidant and free radical
activity of Spondias pinatta
Ervatamia coronaria Stapf. 118.77 India Antioxidant and Free radical
scavenging Activities of
Evartamia coronaria Stapf.
Panax ginseng 3.18 India In vitro evaluation of free
radical scavenging activities of
Panax ginseng and
lagertoemia speciosa
Lagerstroemia speciosa 6.15 India In vitro evaluation of free
radical scavenging activities of
Panax ginseng and
lagertoemia speciosa
Ascorbic Acid 3.35 India In vitro evaluation of free
radical scavenging activities of
Panax ginseng and
lagertoemia speciosa
Basilici herba 57.15 India Assesment of antioxidant
activity of Basili herba
aqueous extract- in vitro
studies
Syzygium jambos* 37.65 Puerto Rico Scavenging and antioxidant
activity of plant extracts
towards superoxide radical
anion
ResearchesAroundtheWorld
6. Relevance of this Research
Pathophysiological Importance
– “Diabetes and obesity are also
associated with the
overproduction of
mitochondrial reactive oxygen
species (Superoxide), leading
to mitochondrial and cellular
oxidative damage.
– This, in turn, contributes to the
development and progression
of diabetic complications and
to worsening of the diabetic
state per se.” William Sivitz,
MD, Endocrinologist
Etnopharmacological Importance
• To study plants with antioxidant and
free radicals scavenging activity has
relevance in the screening of
possible therapeutics for
hyperglycemia and diabetes.
• Anti-diabetic activity of medicinal
plants and its relationship with
their antioxidant property, India,
2003
– Triphala – A mix of thre plants
T.chebula, T.belerica, E. Officinalis.
http://www.free-press-
release.com/news/200803/1206625212.ht
ml
7. 1. Literature Review
Read Scientific Papers of
Researches related to
your area of interest.
2. Do a Protocol of
your Research
How you are going
to do your
Research?
Use the literature
review as
background.
Follow the
Scientific Method.
Concentrations Inhibition
percentage
Scavenging and Antioxidant Activity Towards
Superoxide Radical of Plant Extracts used
as Diabetes Adjuvants
Materials
Methods
3. Follow the
methods
established in the
protocols.
From Protocol to Research: A Description
8. From Protocol to Research: A Description
3. Follow the
methods
established in the
protocols.
4. Data collection &
Data analysis 5. Conclusions
6. Publication
Errors in Data!
Does not work!
Review protocols and
literature to answer:
What happened?
Why it does not work?
9. Methodology:
in vitro Evaluation of Plant Extracts
1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
H
I
Scheme 2-A. The in vitro assay recreates a superoxide radical anions environment.
A B C D E
NBT PMS NADH
85uL 25uL 5uL 25uL
Test
Blanc of the Test
NADH
Blanc
10. Research Progress Step by Step
Literature
Review
Making
Protocols
Data Collection
January 2010 May 2010 January 2011 May 2011 Aug 2011 Oct 2011
Conclusions
Summary
12. Around the world, mostly in India,
several studies have been conducted
to discover good antioxidant and
scavenging plants extracts, in order to
develop new therapeutics to treat
diabetes.
Six plants extracts were found in the
databases of investigations around the
world, showing superoxide radical
scavenging activity.
In our case, S.jambos aqueous and
methanolic extract, showed IC50’s of
37.65g/mL and 79.24g/mL
respectively.
Our results suggest that S.jambos
is a good antioxidant and
therefore, more studies to screen
phytochemicals that scavenge
superoxide radicals is
recommended to develop new
therapeutic methods to treat
diabetes.
IC50 of Diabetes Adjuvants and
superoxide scavengers
Adjuvants around the world IC50’s
Spondia spinatta 13.46μg/ml
Basilici herba 57.15g/mL
Panax ginseng 3.18μg/mL
Lagerstroemia speciosa 6.15μg/mL
Ervatamia coronaria 118.8μg/mL
13. Referencias
1. Tomasz J. Guzik, MD, PhD., Mechanisms of Increased Vascular Superoxide Production in Human Diabetes
Mellitus Role of NAD(P)H Oxidase and Endothelial Nitric Oxide Synthase, 2002
2. Francesco Cosentino, MD PhD., High Glucose Increases Nitric Oxide Synthase Expression and Superoxide
Anion Generation in Human Aortic Endothelial Cells, 2002
3. William Sivitz, MD, Mitochondrial Dysfunction in Obesity and Diabetes, US Endocrinology, 2010;6:20-7
4. Kuniharu Suzumura, Superoxide Anion Scavenging Properties of Fluvastatin and Its Metabolies, 1999
5. C. Hu, Dandelion (Taraxacum officinale) flower extract suppresses both reactive oxygen species and nitric oxide
and prevents lipid oxidation in vitro, 2003
6. Nripendranath Mandal, Antioxidant and free radical scavenging activity of Spondias pinnata, 2008
7. Gavillán-Suárez, J., Alvarado-Guzmán, J. & Germosén-Robineau, L. (2009). TRAMIL ethno-pharmacological
survey: Knowledge distribution of medicine plant use in the southeast region of Puerto Rico. Puerto Rico Health
Sciences Journal, 28(4), 329-339.
8. K Suzumura, M Yasuhara, H Narita, Superoxide anion scavenging properties of fluvastatin and its metabolites.
Chem Pharm Bull (Tokyo) 1999 October
14. Research Skills Developed
Software Applications for Statistical Analysis
SPSS
•Graphic Analysis
•Linear and logarithmic
Regressions
Excel (Microsoft 2007 and For
Macintosh 2011)
•Graphic Analysis
•Linear and logarithmic
Regressions
•Standard Deviations
•Error Bars
•Error Percentage
•Platforms using Java
15. Microplate Reader
pH Meter
Sonicateur Apparatus
Micropipets
Research Skills Developed
Laboratory Equipment used in this Research
16. Interdisciplinary Work
Through all these processes the “Phytochemical and Pharmacognocial lab taught
me:
-Have patience.
-Learn English as a second language
-Writing skills (Essays, Abstracts,
introductions, summaries)
- Do Scientific papers and a poster!
-M.D. or Ph.D. as final career goals.
-To have more confidence on me.
-Share our things. We are a family!
-Be responsible in our stuff.
-Work hard and you will gain success.
-Never gave up!
-The capacity to hear and act.
17. This research experience with Dr. Jannette Gavillán and the Interdisciplinary Research
Institute started in October 2009 and brought me several opportunities:
Interdisciplinary Work
October 2009 October 2010 October 2011
October 2009
Started in the
PPRL and III
January 2010
Started the
Superoxide
and Diabetes
Research
Summer 2010
I went to
North Carolina
Research
Experience
January 2011
NASA
Scholarship
August 2011
PR-LSAMP
Fellowship
November
2011
ABRCMS
Travel
Award
18. Interdisciplinary Work
January 2012
Future Goals in Research
May 2012
March 10
UPR Carolina
PR-LSAMP
32th PR
Interdisciplinary
Meeting
Post-Bac in
University of
Missouri-Columbia
August 2012 August 2013
Graduate School
“Free Radical and
Radiation Biology”
19. This research experience with Dr. Jannette Gavillán and Interdisciplinary Research
Institute started in October 2009 and brought me several opportunities:
Interdisciplinary Work
AMP 6th Transdisciplinary
Research Conference
NC State Summer
Research
Opportunity
Annual Biomedical Research
Conference for Minority Students
Summer 2010 May 2011 November 2011
