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Francoise Sidime Neuroscience PhD Candidate Graduate Center/CSI CUNY Institute of Basic Research (IBR)

Ben Carson – Neuroscientist & Ben Carson – Neuroscientist & neurosurgeonneurosurgeon

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Undergraduate: Class of 2007 Undergraduate: Class of 2007

Secure your GPA – Maintain your Secure your GPA – Maintain your successsuccessSurround yourself with “Like minded

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Pathway to being a Scientist Pathway to being a Scientist (Researcher)(Researcher)Undergraduate (Bachelors): 4 years

Graduate School (Masters): 2 years

Graduate School (PhD): 5 years +

Post Doc

Your network to successYour network to success

• Excellent mentor/PI

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A B C

A=

• High GPA• High GRE Score• Research with publications• Excellent personal statement• Well rounded CV• Excellent letters of recommendation

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Nailing the Interview…….Nailing the Interview…….

Masters - NeuroscienceMasters - Neuroscience2 years course work & thesisEasier than undergrad!Conduct researchTeachCUNY – Delay GRE until PhD

FinancesPay out of pocket: Minority programs help CSTEP, LSAMP, NSF, UNCF-MERCK, HSF

1st year: Take GRE & apply to PhD Programs

PhD - NeurosciencePhD - Neuroscience5 year program: Stipend – No debt!

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Travel to conferences: Present & Network

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Defense: Award PhD in August

What do I do with a PhD?What do I do with a PhD?Post Doc position Professor – University aiming for tenure

Run your own lab/research

Work for pharmaceutical company: Pfizer

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Images compliments of the Marsh-Armstrong Lab - Johns Hopkins/Kennedy Krieger

The El Idrissi Lab - NeuroscienceThe El Idrissi Lab - Neuroscience

The Effects of Gestational Exposure to Low Levels of Di-n-butyl Phthalate in MiceFrancoise Sidime, Irena Jmukhadze, Dinali Obeysekera, Samreen Tariq, Sarah Elkhatib, Dr. Abdeslem El Idrissi

Biology/Neuroscience / College of Staten Island / CUNY, Center Developmental Neuroscience, Staten Island, NY

Abstract

The etiology of autism is thought to involve the complex interplay among genetic and environmental factors. Patterns of inheritance suggest an epigenetic component to the development of autism. A variety of environmental agents are known to induce epigenetic changes in DNA, including those in autism-associated genes. These factors include the plastics by-products DBP and bisphenol A (BPA). To date, although the effect of DBP as an endocrine and a reproductive disruptor are well established, there are only few studies that address the effects of low levels of DBP. Additional research analyzing the low-dose effects of these endocrine disruptors need to be further investigated, since it is most unlikely for humans to be exposed to high concentrations of DBP.

Hypothesis

Data collected and Analysis

Conclusion

References

I would like to thank Dr. Elidrissi for giving me the opportunity to conduct research in his laboratory. In addition I would like to thank Francoise Sidime including the El Idrissi lab for their assistance on the project.

A special thank you goes to the CSTEP office for their support and funding opportunity.

Gender, Genotype, and DBP Effects in Open Field Activity

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Based on our preliminary data, we believe that DBP delays the maturation of the GABAergic system and this delay could be attributed to the inhibition of intrinsic neurosteroids. Inhibition of these neurosteroids early in development will cause neurodevelopmental insults by affecting GABAA receptors. These receptors have a significant role in establishing proper formation and maturation of neuronal networks that are critical for brain development and normal behavior.

Our findings suggest that gestational exposure to DBP will delay the maturation of the GABAergic, and thus delaying normal brain development. As a result, we observed aberrant GABA-dependent behaviors such as increased locomotive activity and elevated anxiety and fear.

Baulieu EE (1991) Neurosteroids: a new brain function. Biol Cell (1991) 71, 3-10

Ben-Ari Y, Gaiarsa JL, Tyzio R, Khazipov R (2007) GABA: A pioneer transmitter that excites immature neurons and generates primitive oscillations. Physiol Rev 87:1215-1284.

El Idrissi A, Boukarrou L, Heany A, Malliaros G, Sangdee C and Neuwirth L. (2008) Effects of taurine on anxiety-like and locomotor behavior of mice. Adv. Exp. Med Biol 643 (7) 207-215.

El Idrissi, A., Boukarrou, L., Heany, W., Malliaros, G., Sangdee, C., &Neuwirth, L.S. (2009).Effects of taurine on anxiety-like and locomotor behavior of mice. In J. Azuma, S.W. Schaffer, & I. Takashi (Eds.), Taurine 7: Taurine for the future healthcare. 643, pp.207-215, New York, NY: Springer Press.

Open Field Test

Elevated Plus Maze

Acknowledgements

Physiology

Behavioral Experiments

RT-PCR experiment

Electrophysiology

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Immunohistochemistry

P450sc StAR Colocalization DAPI

Experimental Design

(A) KO & KO-DBP male mice exhibited an increase in total distance travelled compared to WT

(B) KO & KO-DBP male mice spend more time in the center zone compared to male WT mice

(C) KO & KO-DBP male mice exhibit an increase in total time active compared to WT

(D) KO-DBP mice show a reduction in immobility compared to male WT mice.KO female mice have elevated immobility when compared to WT female mice

These results suggest that DBP treated mice shows pronounced effects of hyperactivity with respect to male KO mice.

Statistical values are representative of a Factor ial ANOVA with Unequal-Tukeys HSD post hoc test alpha level 0.05% (p< 0.05* and p < 0.01**).

(A) KO-DBP male mice entered the open arm less frequently and head exploration (head poke) over the open arm less than WT male mice.

(B) WT-DBP female mice enter the open arm less frequently than all other treatment groups and genotype

(C) WT-DBP & KO- DBP male mice spend less time in the open arm compared to WT male mice

(D) KO & KO-DBP females spend less time in the center zone.

Statistical values are representative of a Factor ial ANOVA with Unequal - Tukey’s HSD post hoc test alpha level 0.05% (p< 0.05*, p < 0.01** and p < 0.001***).

To determine the effects of DBP on early brain development, we injected pregnant mice with DBP (1 mg/kg s.c) on gestational day 10 and assessed the neurobehavioral effects in the offspring when they reached 2 months of age. The importance of administering DBP on gestational day 10 lies in targeting the critical window when neurogenesis occurs between Embryonic Day 10 (E10) and E20. The open field and elevated plus maze will test for anxiety.

Spatial analysis of P450scc, StAR and GABAAR : The following analysis will be conducted by Immunohistochemistry (IHC). For IHC analysis, we will prepare cryosections of controls and fmr1-KO brains offspring of ± DBP at postnatal day 1 and 7, and then examine the expression pattern of P450scc, SARP, SF1, GABAAR, VSCCs and KCC2. These selected developmental times (postnatal day 1 and 7) encompass the time frame of GABA switch from excitatory to inhibitory. In order to define clear comparisons across all treatment groups, all the tissue samples will be subjected to IHC at the same exposure time of 48 hours with the primary antibody, and 1 hour for the secondary antibody prior to being counter stained with an antifade containing DAPI. After completing the IHC, the samples will be imaged and 2-D images of the tissue samples will be collected from the middle section of each sample along with z-stacks of the entire sample for 3-D images. The 3-D digital z-stack will be transferred to the Imaris software where the mean intensity of the fluorescently labeled proteins of interest will be digitally reconstructed and quantified. As the IHC will only confirm the visual presence/absence in the distribution of the protein patterned expression levels, it will be imperative to run both RT-PCR and electrophysiology. RT PCR will also be conducted in order to quantify the key proteins involved in steroidogenesis. Electrophysiology studies including recordings from the cortex will also be conducted in order to access the effect of DBP on the GABAergic system.

DBP Immunoreactivity in the Hippocampus. (A) Stained for the StAR protein(B) Stained for the P450 scc (C) Colocalization

WT KO

WT DBP KO DBP

The Open Field Test shows that DBP has a genotypic and gender effect as the KO & KO-DBP male mice exhibit an increase in total distance travelled compared to the male WT mice. In addition these mice spent more time in the center zone.

The Elevated Plus Maze, a behavioral test that tests for anxiety did not show significant difference in regards to gender, genotype and DBP effect. An additional parameter was analyzed that looked at the mice exploring the outer ledge of the EPM that we called ‘Head Poke.” These results showed a significant difference between the Male WT Mice and the Male KO DBP mice. The Male WT mice spent more time exploring the outer ledge of the EPM compared to the Male KO DBP mice

Pathway to ProminencePathway to Prominence Research opportunities - International

LSAMP ProgramLSAMP ProgramInternational Research 2011– Graz, AustriaInternational Research 2011– Graz, Austria

European Union Bioclaims Project European Union Bioclaims Project Graz - Austria Graz - Austria

Marshall Plan Foundation - 2012Marshall Plan Foundation - 2012International Research in Austria International Research in Austria

MPS Recipients - 2012 MPS Recipients - 2012

The Effects of Ischemia on the Brain - Kidney The Effects of Ischemia on the Brain - Kidney AxisAxis LKHLKH-Univ. Klinikum Graz, Medical University of Graz & TU Graz Male, 6-week old C57BL/6 mice Ischemia by bilateral clamping of renal pedicles (varying duration) Reperfusion Analyze behavior – Open Field & Elevated Plus Maze (EPM)

Ischemia

0 10 min

20 min

30 min

Reperfusion

4h 12h 24h

S-Creat, S/U-Urea, S/U-NGAL, S-Cystatin C, U-

KIM-1

LSAMP New York Delegate - AustriaLSAMP New York Delegate - AustriaTU Graz – Summer Poster Presentation TU Graz – Summer Poster Presentation 20132013

Institute of Medicinal Plants Institute of Medicinal Plants and Science – Taounate, and Science – Taounate, MoroccoMorocco

Institute of Medicinal Plants and Institute of Medicinal Plants and Science – Taounate, MoroccoScience – Taounate, Morocco

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Autism Conference –Rabat, Autism Conference –Rabat, MoroccoMorocco

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Pathway to ProminencePathway to Prominence International Conferences

Taurine Conference 2014- Krakow – Poland Taurine Conference 2014- Krakow – Poland

Krakow – Poland Krakow – Poland ConferenceConference

Cruising the Polish Cruising the Polish ““HudsonHudson””

Maastrich Conference – Maastrich Conference – NetherlandsNetherlands

Taurine Conference 2012 – Taurine Conference 2012 – Marrakesh MoroccoMarrakesh Morocco

Conference in Marrakesh - Conference in Marrakesh - MoroccoMorocco

Conference in Marrakesh-Conference in Marrakesh-MoroccoMorocco

Morocco – Sahara DesertMorocco – Sahara Desert

Pathway to ProminencePathway to Prominence Funding Opportunities

UNCF – MERCK Undergraduate: $25,000 Graduate: $53,000 Post Doctoral: $92,000

Fulbright Scholarship

Hispanic Scholarship Foundation Undergraduate: $500 - $5,000

Marshall Plan Foundation

ConclusionConclusionEarly Preparation is key!Keep your GPA high!Score high on your GREVolunteer!!!Research - InternationalPublishNetwork!!!

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