Abstracts 2016

Resistance Status of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) in Florida

Tashani Brown, Muhammad Haseeb, Lambert Kanga, Whitley Stewart and Tavia Gordo

The genus Aedes is known to be associated with the transmission of the zika virus which has been a major health concern recently. The Asian tiger mosquito (ATM), Aedes albopictus is a serious pest of human and a major vector of yellow fever, dengue fever and chikungunya fever. The yellow fever mosquito (YFM), Aedes aegypti, is known to be a very invasive and serious vector of tropical fevers, including the zika virus. These species are widely distributed in most tropical and sub-tropical areas of the world. The ATM was first discovered in Houston, Texas in August 1985 arriving in shipments of used tires from Japan and Korea (CDC, 1986). Currently, it is widely distributed in number of southeastern, and northeastern states in the United States. Immature ATMs and YFM inhibit many different types of containers, and scrap tires which harbor mosquito more frequently and in greater numbers than any other mosquito species. Pesticides are usually used to control these serious pests, however, due to resistance development in several synthetic chemical compounds it is getting difficult to manage it properly. Malathion and permethrin are two of the frequently used pesticides in urban environment to control mosquitoes in Florida. The best possible approach to manage this serious pest is to control pesticide resistance. The current study is being conducted to monitor the resistance of Malathion and permethrin Aedes albopictus and Aedes aegypti under the laboratory conditions.

The Role of Midasin in Letrozole Sensitive Breast Cancer Cells

Tonja Bryant1, Michael Davidson2, and Syreeta L. Tilghman2

While currently women with estrogen-receptor positive breast cancer are treated with anti-estrogens or aromatase inhibitors, resistance to aromatase inhibitors (AIs) (such as letrozole) is a lingering problem which could lead to metastasis and death. There are no cures for metastatic breast cancer so it is critical to understand novel mechanisms contributing to the progression of AI-resistant metastatic breast cancer. Recently, we generated preliminary data that links midasin, a novel nuclear chaperone protein involved in ribosomal biogenesis and export, to increased stemness and the metastatic phenotype in endocrine resistant breast cancer.  Here, we hypothesize that AI-resistant breast cancer is associated with increased midasin expression. Therefore, the overall purpose of this project is to determine whether midasin is expressed differently in letrozole sensitive breast cancer cells versus letrozole resistant breast cancer cells. In order to interrogate this hypothesis immunofluorescence staining of letrozole sensitive breast cancer cells was performed.  Results demonstrated that midasin is expressed in the nucleus and the cytoplasm of AI sensitive cells. Taken together, these results provide a rationale to study the role of midasin in AI-resistance breast cancer, and whether midasin could serve as a novel target for aggressive breast cancer.

Water Drainage in Two Areas of the Ochlocknee River Watershed in Florida

Marquise Cromartie1, Dr. Elijah Johnson

An analysis of the Ochlocknee River was conducted to determine to location of stream gages  that will assist  in our  goal of preparing a model of the Ochlocknee River watershed. This model will demonstrate to us the quality of the calculated water flow results in comparison with the measured water flows.

Acid Stress Response In Various Aquatic Bacteria Using Bioinformatics

Gregory K. Greene1, Mitchell O. Roth, Richard A. Long2

Bioinformatics is transforming data into useful knowledge. In this research we applied various Bioinformatics and data mining techniques. We examined the water borne pathogen Vibrio cholerae in water samples, testing for Acid stress response genes or characteristics.

Water drainage in the Carrabelle River Watershed

Mark Guthrie, Dr. Elijah Johnson, Dr. Edward Ofori

A Watershed is an integral part of the environment. It is nature’s drainage system and an important piece to underground water flow. Due to the ever growing cost of environmental research, we utilize computer simulation software such as Basins to help identify problems within a selected area watershed. Watershed modeling simulates the watershed by use of meteorological data and geographical mapping to reveal an image of the selected area (refer to Figure 3). The data obtained was analyzed using WdmUtil and will be optimized using PEST. Both are software tools existent within the Basins software suite. We opine that sediment testing will reveal the amount of nitrates that are present in the water flow and how much nitrates is coming in and out of the watershed.

Identification and Comparison of the Spotted Wing Drosophila, Drosophila suzukii and the African Fig Fly, Zaprionus indianus (Diptera: Drosophilidae) in North Florida

Sharise James, Muhammad Haseeb, Tavia Gordon and Dasia Harmon

The Spotted Wing Drosophila (SWD), Drosophila suzukii and the African fig fly (AFF), Zaprionus indianus are both invasive pests of soft-skined fruits including blueberries. During the experiment, blueberries were collected from the Center for Viticulture & Small Fruits Research. Collection was conducted for a period of four weeks and observations were made up to the six weeks. Both species emerged from the infested fruits. During those weeks a high population (80-90%) of adult Z. indianus emerged rather than the D. suzukii. Molecular analysis was performed to further confirm the identification of the D. suzukii and Z. indianus. Molecular analysis study was only conducted to gel electrophoresis due the limited time. The gel electrophoresis showed D. suzukii DNA present in the gel, but the Z. indianus DNA was not present in the gel. Generally, Z. indianus is considered a secondary pest of blueberries, however, D. suzukii is a primary pest. The species lays its eggs in fresh soft skinned fruits and later causes economical damage to blueberries.

Synthesis of Steroidal Estrogens as Anti-Breast Cancer Agents

Gail Lopez, Linda Mbiza, Riccardo Jean, Patrick Joseph Ph.D., John S. Cooperwood Ph.D.

Tamoxifen (TAM) is the most common drug used to treat estrogen-dependent (ER+) breast cancer, but certain drawbacks are evident including drug resistance (breast cancer cells become resistant to TAM within 5 years), tumor progression, patient relapse, as well as risk of developing endometrial cancer (JNCl J Natl Cancer Inst (1994) 86). Because of this, there remains a need to pursue alternative treatment options. By synthesizing steroidal estrogen-derivative drugs, the hope is to develop a drug that will effectually treat breast cancer while lessening potential risks. Cytotoxicity studies and proliferation screenings were carried out on MDA-MB-468 cancer cell line with 10% FBS (normal media) and on MDA-MB-468 with 2.5% FBS media. The stock solution was prepared at 20 mg/ml and diluted serially to obtain concentrations up to 250 μl/ml with 6,000 cells/well for proliferation screenings and 40,000 cells/well for cytotoxicity studies. The IC50 values were generated using OriginPro. Compounds 1 and 2 showed highest anti-cancer effects out of the 7 drugs tested.

In Vitro Analysis of Newly Developed Compounds as Potential Anticancer Agents: Inhibition of LNCAP Prostate Cancer cells

Mayfield Sayvion, Carl B. Goodman, Riccardo Jean, Linda Mbiza

Prostatic carcinoma (PC) is one of the most diagnosed cancers in men worldwide. The prognosis of PC is higher in developed countries and amongst individuals of African descent; two-hundred and seventy five per one hundred thousand of black men are affected globally (Crawford). The essential mechanisms of this disease includes the dependence of androgens and gradual independence of androgens over time. The current and limited treatment options have not alleviated the issues of PC. PURPOSE: Using LNCAP cell lines, we sought to determine if these cytostatic compounds support potential anticarcinogen or antiproliferative properties. The compounds were also screened on triple negative breast cancer for effectiveness.  METHODS: Standard mammalian sub-culturing, trypan blue dye exclusion assay, proliferation screening and cytotoxicity assay were used to determine cell viability amongst six compounds from the drug discovery core. RESULTS: After 7 days of drug exposure, the IC50 of compounds GM 243, GM 225, GM 267, and GM 247 showed antiproliferative behavior below 20 μM. CONCULSION: The compounds showed growth inhibition with an IC50 < 20 μM and possess antiproliferative properties at our targeted molar mass. Considering the potential at their efficient molar mass for drug development, the compounds grant leeway for further research.

In-vitro analysis of newly synthesized compounds as potential Triple Negative breast cancer therapeutic agents

Linda Mbiza, Dr. Carl Goodman, Riccardo Jean, Sayvion Mayfield,  Gail Lopez

Triple Negative Breast Cancer (TNBC) is a biologically aggressive subtype of breast cancer that accounts for 10-15% of all breast cancer cases. TNBC lacks estrogen, progesterone, and HER2 receptors, therefore ruling out hormone therapy and Herceptin as effective treatments. Current treatments for TNBC include chemotherapy, radiotherapy and surgery. The PURPOSE of this research is utilizing the Drug Discovery Core (DDC) to analyze compounds and their mechanisms of action on TNBC while determining if there is potential for a treatment. By screening a catalog of microtubule inhibitors and steroidal anti-estrogen compounds on the TNBC cell line, MDA-MB-468 (African American), the most efficacious compound will be determine and evaluated further. The RESULTS concluded that the steroidal anti-estrogen compounds, GL-I-30 and GL-VI-35 were the most potent at the lowest concentration.  These compounds will then be analyzed further, in FUTURE RESEARCH, for the effects on replicative DNA synthesis, tubulin polymerization, cytoskeletal morphology, mitotic index, and distribution of DNA in cell cycle phases.

Synthesis of Potential Antipsychotics

Elishia Pla, Dr. Seth Y. Ablordeppey, Dr. Edward Ofori

The pharamacotherapy of Schizophrenia relies on antipsychotics which elicit their effects by binding to dopamine and serotonin receptor subtypes. Previous work from our laboratory identified lead compound 1 as ligand with multi-receptor binding profile consistent with our working hypothesis. In this work, we used commercially available starting materials to synthesize an indanone analog of 1 to enable us further explore the structure-affinity-relationships (SAFFR).

Design and control of a four bar simulator

Deshon Purvis, Dr. Carl Moore, Twan Capehart, Isaac Odaf

In this project the goal is to develop an easy to use four-bar simulator for instructional purposes. The four-bar is one of the most useful mechanisms in engineering, but is typically viewed and taught in 2 dimensions. To better understand how four-bar linkages can be developed and used this project aims to make a simulator that is; animated, and displays information such as angular position and velocity. The simulator was constructed using: wooden dowel, plastic joints, DC motor, and an accelerometer chip. The simulator will be paired with both National instruments Lab View soft wear, and a USB-6001 DAQmx to control the animation, and read data from the accelerometer. The principle point of this project is to construct an integration based algorithm that is capable of using the data of the accelerometer, and produce outputs such as angler position and velocity.

Targeting human DNAJAs for sensitization to chemotherapeutic agents

Rebeca Rodriguez, Dr. Flores-Rozas, Aurellia Whitmore

Doxorubicin is a drug used to treat over 60 different cancers and it is the first line of treatment for Triple Negative Breast Cancer (TNBC). Doxorubicin’s well known mechanism of actions (MOA) include: intercalation into the DNA, production of reactive oxygen species (ROS) and inhibition of topoisomerase II, which results in cell death. Doxorubicin is a dose dependent type of standard therapy that also affects normal cells, especially cardiomyocytes when high doses are administered for effective therapy. Our goal is to find a way to sensitize cancer cells by targeting the Heat Shock Proteins (HSPs) DNAJAs, that will result in a lower administered dose reducing the side effects associated with Doxorubicin. To test this we examined single-gene deletion strains (Δ mutant) with Etoposide and Menadione. We discovered that inactivating DNAJA1 and DNAJA2 result in most sensitivity to the drugs. As expected all strains were resistant to etoposide.  Therefore, DNAJA1 and DNAJA2 play an important role in the resistance to chemotherapy with doxorubicin and Menadione respectively. Future works includes the use of these cytotoxic agents with TNBC cell lines.

Ecological Comparison of Various Aquatic CRISPR bacteria using data mining techniques

Mitchell O. Roth1, Gregory K. Greene, Richard A. Long2

This research focuses on how different data mining techniques can be utilized to visualize and understand patterns within the data. Specifically, we will be analyzing the relation of CRISPR/Cas immunity with two different trophic environments (oligotrophic & copiotrophic).

Neuroprotective Effects of Thymoquinone and Umbilical Cord Stem Cells

Ian Schlander, Dr. T. Womble

In the present study, PC-12 cells are exposed to hydrogen peroxide induce oxidative stress induced cell death.  This is a cell model for neurodegeneration. Previous to exposure, the cells are treated with thymoquinone, a compound that shows promising neuroprotective qualities, while also exposed to umbilical cord stem cells for its regenerative qualities in order to study the combination of their neuroprotective characteristics. The cells displayed an increase in confluency when given the pretreatment and exposed to oxidative stress compared to the samples exposed to only the hydrogen peroxide. Our results indicated that the combination of thymoquinone and umbilical cord stem cells lead to an increase of each other’s neuroprotection on the cells, offering an encouraging new approach to alleviating  neurodegeneration.

Contaminants, Wastewater and Consequences

Keanu E. Snow, Dr. E.E. Kalu, Shannon Anderson, & Dr. Edward Ofori

Ibuprofen (IBP) concentration is abnormally high in wastewater and its treatment is necessary to alleviate threats to biological communities. Recognition of this has created interest within the scientific community to treat this compound at its varying concentrations, no matter how minute. Zero Valent Iron (ZVI) has been shown to be an environmentally friendly and efficient way of treating this compound. In this work the catalyst(s) were prepared using two methods: Electroless deposition and green chemistry. Their effectiveness in treating Ibuprofen was tested in a batch reaction setup.

The electrolessly deposited ZVI was more reactive in the degraded IBP solution. Additionally, the concentration of the IBP solution was positively influential within the catalytic response. Degradation rate improved within acidic rather than alkaline conditions. Results of ongoing research aim to improve the ability to implement treatment technology into wastewater bodies.

Maintaining Stability: Tip-over Prevention

Marc Toussaint, Dr.Carl A. Moore, Isaac Odaf

In this project, the focus consists of preventing robot tip-over during teleoperation. This includes using an onboard arm to actively prevent tip-over and designing algorithms to prevent the arm from placing the robot in a position to tip-over.

A Novel Nuclear Chaperone Protein is Associated with Endocrine Resistant Breast Cancer

Boyada Tep1, Michael Davidson2 and Syreeta L. Tilghman2

Aromatase inhibitors (AI), like letrozole, are the first-line treatment for estrogen receptor positive breast cancer in post-menopausal women.  Despite widespread successful usage of letrozole, for the treatment of breast cancer, resistance to therapy, tumor relapse, and metastasis remain the principal causes of death for breast cancer patients. While understudied, several reports suggest that misregulation of signaling pathways in cancer cells stimulate ribosome biogenesis, and conversely, defects in ribosome assembly can lead to disease. Recently, we generated preliminary data that links midasin, a novel nuclear chaperone protein involved in ribosomal biogenesis and export, to increased stemness and the metastatic phenotype in endocrine resistant breast cancer.  Currently, targeted approaches to endocrine resistant breast cancer are lacking due to the inability to completely unravel the rate limiting proteins and pathways that are essential to metastatic disease.  Therefore, there is a critical need to identify novel targets for metastatic breast cancer that can prevent recurrence and mortality. Unless such targets are found, treatment options for women diagnosed with endocrine resistant, metastatic disease will be limited to toxic chemotherapy and radiation, both of which are unlikely to cause remission ultimately leading to increased mortality. We hypothesize that overexpression of midasin facilitates endocrine resistance by altering ribosomal biogenesis and contributes to the increased cancer stem cell population.  In order to address this hypothesis, midasin protein expression was examined in long term letrozole sensitive human breast cancer cells (AC-1) cells by immunofluorescence staining.  Results demonstrated that AC-1 cells stained positive for midasin in both the nucleus and cytoplasm.  These results suggest that midasin may be critical to the growth of AI-sensitive breast cancer, however future work is required to demonstrate the role of midasin in endocrine resistant breast cancer.

Abstracts 2015

Functional Analysis of Proteins in Breast Cancer Cell Lines Treated with Grape Extracts

Tonya Harris1 *, Ramesh Katam2, Varshini Sridhar2, Keyura Katam2, Carl B Goodman1

1College of Pharmacy and Pharmaceutical Sciences, 2Department of Biological Sciences, Florida A&M University, Tallahassee FL

Plant extracts have shown potential bioactive compounds that are related to many health benefits. Muscadine grapes contain polyphenol compounds that showed potential antioxidant and anticancer properties. Although various compounds exhibit the specific activity, molecular mechanisms in cancer cell lines treated with the plant extracts have not been fully understood. Hence, the goal of this research is to determine the pathway of proteins associated with anticancer activity in breast cancer cell lines treated with muscadine grape extracts. Gradient concentrations of grape pericarp, and seed extracts were tested on human breast (MDA-MB-231) cancer cell line to test cancer cell growth inhibition. Cytotoxicity of MDA-MB231 cells, which were maintained with RPMI-1640 media, was measured with crystal violet assay. The results showed significant cell death of the cancer cells treated with pericarp extract. Total proteins were extracted from cells using lysis buffer solution. Proteins were resolved on two-dimensional electrophoresis and protein spot densities were analyzed using PD Quest. Differentially expressed proteins were characterized on MALDI/TOF. Proteins were identified using MASCOT algorithm. Functional annotation was carried out using bioinformatics tools to determine the role of these proteins in the metabolic pathways. A total of 35 proteins, specific to anti-cancer activity, are differentially expressed in breast cancer cell line treated with grape extract. Six proteins are up regulated and twenty-nine proteins are down regulated. Up-regulated proteins molecular functions mainly involve binding and structural molecule activity, which plays a role in the regulation of anticancer activity. Down-regulated protein groups is mainly involved in cellular process, signaling, and developmental process, which provides information for the mechanisms of anticancer effect of grape extract. Findings suggest that proteins involved in cytoskeleton, cell signaling, and cellular integrity are

Malathion Resistance Status of Aedes albopictus in Leon County, Florida

Tashani Brown, Whitley Stewart, Muhammad Haseeb and Lambert Kanga

Florida A&M University Bridges to the Baccalaureate in the Biomedical Sciences,
College of Agriculture and Food Sciences, Tallahassee, FL

Asian tiger mosquito, Aedes albopictus is a serious pest of human and a major vector of yellow fever, dengue fever and chikungunya fever. The species is native to Southeast Asia. It is widely distributed in most of tropical and sub-tropical areas of the world. First discovered in Houston, Texas, in August 1985 arriving in shipments of used tires from Japan and Korea (CDC, 1986). Currently, it is widely distributed in number of southeastern, and northeastern states in the United States. Pesticides are usually used to control this serious pest, however, due to resistance development in several synthetic chemical compounds it is getting difficult to manage it properly. Malathion is one of the frequently used pesticides in urban environment control mosquitoes. This preliminary study was undertaken to monitor Malathion resistance in A. albopictus under the laboratory conditions.

Synthesis and biological evaluation of 1-(pyridin-2-yl)piperazine derivates as potential antipsychotics

Imani Morris1, E Ofori2, and Seth Y. Ablordeppey3

Florida A&M University, Tallahassee, FL

Bridges to the Baccalaureate in the Biomedical Sciences

 

Seventy million people worldwide live with schizophrenia and out of that population 80% of them stop taking antipsychotic treatment due to ineffectiveness and intolerable side effects. Schizophrenia presents as positive, negative, and cognitive symptoms. Antipsychotics are classified into typical and atypical depending on their tendency to cause extrapyramidal side effects which is seen principally in the latter. Aside dopamine, the atypicals bind to serotonin receptors which lead to reduction in motor side effects. Even though the atypical antipsychotics treat some of the negative symptoms they fail to treat the cognitive impairment. There is therefore a need to search for effective and safer antipsychotics. The current paradigm in antipsychotic drug design is to design small molecules which trigger β-arrestin signaling. Functionally selective ligands which trigger signaling at the β-arrestin pathway have been found to be more efficacious and have lesser side effects. Using structure-activity relationship studies our lab has identified a lead compound 1-(4-(4-fluorophenyl) butyl)-4-(pyridin-2-yl) piperazine (1) which has demonstrated functional selectivity (92.92%) at β-arrestin pathway of the dopamine-2 receptor (D2R) using a Tango assay. In an attempt to better understand the Structure-Functional Selectivity Relationship, we explored the various parts of the lead molecule. To this end, we have successfully synthesized derivatives of the molecule with particular focus on the phenyl left hand side. The exploration is still in progress.

Prodrugs of Steroidal Anti-Breast cancer Agents

Richard Hudson, Patrick Joseph and John S. Cooperwood

Florida A&M University College of Pharmacy and Pharmaceutical Sciences Tallahassee, FL 32307

BACKGROUND: Estradiol (E2) is important for the development of the female reproductive system and female characteristics, but it is also able to cause breast cancer to spread by binding to the estrogen receptor. Tamoxifen (TAM) is the most common drug used in the treatment of estrogen-dependent breast cancer cells.  It works by binding to the estrogen receptor in order to prevent estradiol (E2) from stimulating the growth of breast cancer cells.  Breast cancer cells become resistant to TAM within five years . Therefore we need to develop alternative agents.  We hope to accomplish this objective by synthesizing ester prodrugs of known active steroidal anti-breast cancer agents.  OBJECTIVE: By synthesizing prodrugs of active anti-estrogen steroids, we will increase potency due to enhance lipophilicity; thereby improving cell penetration. By improving cell penetration, we hope to allow a higher concentration of a compound to be available to elicit an action.   By displaying a greater potency, the likelihood of resistance will be reduced due to the cytotoxic effect. METHODS: We followed established chemical procedures in order to modify the structure of  E2 and obtain various estradiol derivatives.  Various basic side chains were attached to the hydroxyl group of the A-ring follow by esterification of the hydroxyl group of the D-ring using a well known procedure RESULTS: The estradiol derivatives were obtained in good yields and successfully identified using  Proton (H) Nuclear Magnetic Resonance (NMR).  The Log P (log of partition coefficient) was determined using Percepta which show an enhancement in lipid solubility CONCLUSION: We successfully synthesized steroidal derivatives which computationally showed higher Log P in comparison with parent active anti-breast cancer agents.  These steroidal derivatives will be further studied against MCF-7 and MDA-MB-231 breast cancer cell lines.

The Antiproliferative Effects of Curcumin on MDA-MB-231 Human Breast Cancer Cells

Kyra Morgan1,2,6* , Timonia Buchanan1,3,4 , Tray Womble1,2 , Jesse Edwards1,3,5 , Donald E. Palm1,2 , LeeShawn D. Thomas1,3,4*

1Florida A&M University, Tallahassee, Florida, United States, 32307 2FAMU College of Pharmacy & Pharmaceutical Sciences 3FAMU College of Science & Technology, 4FAMU Department of Biological Sciences, 5FAMU Department of Chemistry 6Tallahassee Community College, Tallahassee Florida, United States, 32304.

Abstract

Cancer is commonly defined as the unrestrained growth of malignant cells that can occur anywhere in the human body. Nutrition, genetics, lack of exercise, smoking, alcohol, aging, viral infection and adoption of the western lifestyle are contributing factors in cancer. Cancer is a multistep process that involves: (1) initiation (genetic damage), (2) promotion (proliferation of mutated cells), and (3) progression (metastasis of mutated cells). Breast cancer is the second leading cause of cancer death in women where 1 in 36 women will develop breast cancer. While there are various types of breast cancers, the cell line that was used in this research was MDA-MB-231, a triple negative breast cancer (TNBC) cell line. In triple negative breast cancer the estrogen, progesterone, and human epidermal growth factor receptors are all negative meaning, they do not play a role in the growth and development of the cancer. TNBC is one of the more aggressive types of breast cancer which makes it more difficult to treat. Because of the serious side effects, toxicity and resistance of synthetic chemotherapeutic drugs in cancer, science and medicine are investigating natural endogenous chemicals in plants as anti-cancer agents and/or adjuvant to chemotherapy. The use of nutraceuticals as chemopreventive reagents has become popular in in-vitro studies of their biological effects in cultured human cells. Nutraceuticals (hybrid of ‘nutrition’ and ‘pharmaceutical’) are foods or components of food that play a significant role in modifying and maintaining normal physiological functions to keep the body healthy. Research shows that nutraceuticals have been proven to reduce malignant calls in breast cancer and other types of cancer. The nutraceutical used in this research was Curcumin, the main active ingredient found in Turmeric. Curcumin has become a therapeutic supplement that is being used to treat a wide variety of conditions and diseases, such as breast cancer because it has an ability to kill cancerous cells without harming normal ones. The TNBC cells were used as an in vitro model to mimic some of the biological effects of breast cancer. The cells were grown to 80% confluencey in Dulbecco’s Modified Eagle’s Medium (DMEM), 10% Fetal Bovine Serum (FBS), 1% Penicillin-Streptomycin and were incubated at 37˚C with 5% CO2. Next, the MDA-MB-231 cells were treated with 1% DMSO (Control) or Curcumin (12.5 – 200mM) for 24 or 48 hours before being assayed for cell viability or visualized using light microscopy. Then, the cells were assayed for cell viability using Resazurin 10-20% (v/v) and incubated in 95% air and 5% CO2 in a humidified incubator at 37°C for 3 hours. After having 24 hours of Curcumin exposure, results showed that the Control had 100% cell viability, 12.5µM of Curcumin had 97.98%, 25µM had 79.99%*, 50µM had 61.45%*, 100µM had 34.35%*, and 200µM had 13.76%*. After 48 hours of exposure data showed the Control had 100% cell viability, 12.5µM of Curcumin had 75.73%, 25µM had 35.43%*, 50µM had 21.75%*, 100µM had 11.00%*, and 200µM had 8.68%*. The results from this study indicate that: (1) cell proliferation is significantly decreased in a dose and time dependent manner; (2) the morphology of cells is modulated in a dose and time dependent manner in comparison to cells only treated with the vehicle (DMSO); (3) the morphological changes are possibly the result of curcumin’s direct or indirect actions on cytoskeletal genes which are necessary for attachment, anchorage and invasion.

Proteomic Analysis of the effects of Thymoquinone in Oxidative Stress induced Mouse Neuroblastoma cells

Godwin Denyo, Matthew Clowers ,Priscilla K. Medehue,  Tracy A. Womble

Florida Agriculture and Mechanical University, College of Pharmacy and Pharmaceutical Sciences, Bridges to the Baccalaureate in the Biomedical Sciences, Tallahassee, Florida 32307

 

Thymoquinone (TQ) is a phytochemical compound found in black seed oil (Nigella Sativa) with anti-inflammatory effects and the power to inhibit tumor cell proliferation. It is effective against cardiovascular complications, diabetes, asthma, kidney disease, cancer etc. Oxidative stress is an imbalance between the production of free radicals and the ability of the body to counteract or detoxify their harmful effects through neutralization by antioxidants. Oxidative stress can lead to many pathophysiological conditions in the body including Parkinson’s, Alzheimer’s, gene mutations, cancers, chronic fatigue syndrome, and heart attacks. We investigated the neuroprotective effect of TQ following oxidative stress in N2A cells.

Inhibition of Polyisoprenylated Methylated Protein Methyl Esterase (PMPMEase) Induces Cell Death and Inhibits Cell Migration in Lung Cancer Cells

Jerrine Fletcher, Elizabeth Ntantie Ph.D., and Nazarius S. Lamango PhD

 

FAMU Bridges to the Baccalaureate in the Biomedical Sciences

Florida A&M University

College of Pharmacy and Pharmaceutical Sciences

Tallahassee, Florida 32307

 

Despite recent advances in biomarker discovery and targeted drug development, lung cancer still accounts for 27% of cancer-related deaths in the United States, resulting in an estimated 158,040 deaths in the US in 2015. We previously observed that polyisoprenylated-methylated-protein-methyl-esterase (PMPMEase) is overexpressed and hyperactive in lung cancer tissues and cells. Since polyisoprenylation is essential for the oncogenic effects of proteins such as Ras, we have developed polyisoprenylated cysteinyl amide inhibitors (PCAIs) as putative PMPMEase inhibitors for cancer therapy. We hypothesized that one of the PCAIs, NSL-BA-040, will induce lung cancer cell death and inhibits lung cancer cell proliferation, migration, and survival. Using cell viability, cell proliferation, wound healing and colony formation assays, the effect of NSL-BA-040 on H460 lung cancer cells was investigated. NSL-BA-040 induces apoptotic cell death with an EC50 of 4.2 µM, which is more potent than docetaxel, paclitaxel, and erlotinib with EC50 values of 56.4 µM, 134.5 µM, and >200 µM, respectively. NSL-BA-040 significantly suppresses H460 cell proliferation and migration at concentrations as low as 0.5 µM, and suppresses H460 cell survival by 30 % at 2 µM. Results indicate that the PCAI, NSL-BA-040, thus has the potential to be developed as a new class of effective and targeted therapy for lung cancer.

Farnesol inhibition of viability and migration of cancer cells: Implications for prostate cancer prevention and therapy

Kehinde Idowu, Felix Amissah, Augustine Nkembo, Rosemary Poku, Nazarius S. Lamango

College of Pharmacy and Pharmaceutical Sciences, Florida A&M University Tallahassee, Florida 32307

FAMU Bridges to the Baccalaureate in the Biomedical Sciences

Background: Flavors are compounds found in foods and may act as chemical attractants or deterrents. Flavors such as Farnesol have been associated with cancer prevention.

Purpose: To determine the effects of Farnesol and trans, trans-farnesol on

  • The viability of Mia-PaCa-2 cells
  • Mia-PaCa-2 cell migration
  • F-actin filaments organization in Mia-PaCa-2 cells
  • PMPMEase activity in Mia-PaCa-2 cells

Methods: Cultured Mia-PaCa-2 cells were treated with both flavors (0.1 – 10 µM) and analyzed for their effects on F-actin organization, cell viability, cell migration and apoptosis.

Results: Treatment of Mia-PaCa-2 cells with Farnesol and trans,trans-farnesol were observed to be most effective between concentrations of (1-10 µM) and caused apoptosis of cells.

Conclusion: These compounds found in nutraceuticals have the ability to induce apoptotic cell death and inhibit cell migration, which might be an effective therapeutic agent against the fight of pancreatic cells.

Characterization of Chronic Morphine Treated C6 Glial Cells

Maraina Monroe1, Kesa Randell2, and Carl B. Goodman2

FAMU Bridges to the Baccalaureate in the Biomedical Sciences

Tallahassee Community College1, Florida A&M University2, College of Pharmacy and Pharmaceutical Sciences, Tallahassee, Florida 32307

Background: Morphine is the most common opioid used to treat chronic and acute pain in a clinical setting. When morphine is taken over an extended period of time, a phenomenon known as morphine tolerance develops. This opioid acts on the mu opioid receptor, specifically mu1 when looking at its analgesic properties, therefore morphine is considered an agonist at the mu opioid receptor. The drug naloxone is an antagonist to the mu opioid receptor, and will therefore be used to see if any effects that are observed by morphine exposure are mu receptor mediated.  Purpose: To determine what role glial cells play in the development of morphine tolerance.  The present study was designed to investigate the effect of morphine on cell viability and its role in regulating the expression of important cytokine genes.  Methods:  Cells were exposed for 1 hr or 24 hrs to 0, 1, 5, 10, 15, and 20 μM of morphine, naloxone, or 1 hour pretreatment with naloxone followed by morphine.  After drug exposure, cell viability was  examined using alamar blue.  Based on cell viability results and previous literature, cells were exposed to 10 μM of morphine for 24 hrs.  Following drug treatment, cytokine gene expression was assessed using PCR array.  Results: One hour time point cell viability studies showed a significant decrease in cell viability at the 5 uM and 20 uM morphine treatment as well as the 20 uM  nalxone pretreatment (1 hr) + morphine.  After 24 hr morphine exposure, the cell viability assay indicated significant decrease in viability across all treatment groups.  Following 24 hr morphine exposure (10 μM), expression of aminoacyl tRNA synthetase complex-interacting multifunctional protein 1 (Aimp1), colony stimulating factor 1 (macrophage) (Csf1), macrophage migration inhibitory factor (Mif), nicotinamide phosphoribosyltransferase (Nampt), secreted phosphoprotein 1 (Spp1), transforming growth factor, beta 1 (Tgfb1), transforming growth factor, beta 2 (Tgfb2), and vascular endothelial growth factor A (Vegfa) were increased compared to the control.    Conclusion:  These findings suggest that chronic morphine treatment effects glial cell viability and cytokine expression which may contribute to the development of morphine tolerance.

Cocaine Induced Astrocyte-Mediated Expression of Pro-Inflammatory Cytokines VEGF and IL-6

Shamar Banks1, Marquita Johnson2, and Carl B. Goodman2

 FAMU Bridges to the Baccalaureate in the Biomedical Sciences

Tallahassee Community College1 and Florida Agricultural & Mechanical University2, College of Pharmacy and Pharmaceutical Sciences, Neuroscience Division

Tallahassee, Florida 32307

Background: Cocaine is a potent psychoactive compound that is classified as a schedule II drug that exhibits a high risk for abuse but also has some accepted medical value. In the United States, cocaine is the 3rd most abused drug. Long-term cocaine abuse has been associated with neurodegenerative disorders such as Alzheimer’s disease (AD), Parkinson’s disease (PD), Amyotrophic Lateral Sclerosis (ALS), and early onset dementia. Disease progression has been linked to the dysfunction of astrocyte morphology, and chemical signaling, specifically the over expression of pro-inflammatory cytokines Vascular Endothelial Growth Factor (VEGF) and Interleukin-6 (IL-6). Purpose: The purpose of this investigation is to determine the biochemical significance of astrocyte-mediated expression of VEGF and IL-6 following chronic cocaine treatment. Methods: Rodent C6 astroglioma cells were treated with sub-lethal doses of cocaine at 2mM, 3mM and 4mM concentrations for 24 and 48 hours. Western Blot analysis was used to determine the relative expression of VEGF and IL-6 in the control and cocaine treated astrocyte cultures. Results: Cytotoxicity studies showed that cocaine should a dose and time dependent effects in the C6 glial cells viability.  Western blot analysis and ELISA studies were conducted to show a correlation with cytotoxicity and VEGF ad IL-6 expression. Conclusion: The data obtained from this study will provide a better understanding the biochemical basis of cocaine abuse in astrocyte function and cocaine-mediated onset of neurological disorders.

Metagenomic Analysis of Soil Bacterial Diversity at a Geothermal Hot Spring, Manikaran, India

Cason Knight1, Gregory Hitz2, Ashvini Chauhan2

1FAMU Bridges to the Baccalaureate in the Biomedical Sciences, Florida A&M University

2School of the Environment, Florida A&M University, Tallahassee, FL-32307

It is estimated that 99% of the microbes have not yet been cultured. Metagenomics refers to the study of genomic DNA obtained from microorganisms that cannot be cultured in the laboratory. Metagenomics analyses genomic information (16S rRNA gene amplification) and provides an opportunity to investigate the vast majority of life on Earth. Thus, metagenomics plays a crucial role in allowing us to examine DNA from all microorganisms within a community, versus a single organism.

Geothermal springs harbor thermophilic and hyperthermophilic microorganisms; some of which can possess valuable industrial enzymes (Batra et. al. 2011). Specifically, Manikaran springs are typically 96°C but in the surrounding soils, which are continuously saturated by the spring water and have a mousse like appearance, temperatures oscillates between 50°C-60°C (Pandey 1995). These springs were investigate regarding the species diversity in the surrounding soil-mousse.

Synergism of Serine and Tryptophan Metabolic Intermediates and Anthracycline Antibiotics

Celida Brow-Hendrickson, Hernan Flores-Rozas, Jana S. Miles

Anthracyclines constitute drugs of choice as single agents or in combination for the treatment of refractory cancers.  For example, anthracycline are the main therapeutic alternative in triple negative breast cancer, which is commonly observed in African-American women (AA) and is characterized for lacking biomarkers for targeted therapy, and for being aggressive and carrying bad prognoses. Anthracyclines effectiveness is dose-dependent, which also increases the risk of side-effects and eventually the development of drug resistance. In S. cerevisiae, mutants of TRP1, SER1, and SER2 display hypersensitivity to doxorubicin by inactivation of the N-(5’-phosphoribosyl)-anthranilate isomerase, 3-phospho-hydroxypyruvate, and3-phospho-serine, respectively.  We hypnotized that the accumulation of corresponding metabolites at these steps, sensitizes the cells to anthracyclines.  We determined the sensitivity of the yeast strains trp1, ser1, and ser2 to doxorubicin (20 πM) and hydrogen peroxide (3mM). The survival of the trp1 strain was 21% after exposure to doxorubicin (20 πM) which was lower than that of the wild type (37%), while the survival of ser2 (6%) was 6-fold lower. When the strains were exposed to hydrogen peroxide (3 mM), the viability of the trp1 was 6% of ser2 (37%) and ser1 (33%). All below the survival of the wild type (42%) at this dose. The potential use of tryptophan and serine metabolic intermediates along with cytotoxic drugs could potentially lead to application intended to enhance anthracycline-based therapy.

Acid Stress Response in Aquatic Environmental Bacteria

Chelsea R. Hernandez1 and Richard A. Long2

1FAMU Bridges to the Baccalaureate in the Biomedical Sciences

2Department of Biological Sciences, College of Science and Technology

Florida A&M University

Tallahassee, Florida 32307

Enteric bacteria are of interest to the studies of pathogenic microbial ecology for its ability to tolerate high acidic levels stress. Previous research on Acid stress response (ASR) in enteric bacteria have shown that pre-conditioning in mildly acidic pH results in greater survival rates when the bacteria is introduced to very acidic environments, such as the stomach. Which links to increasing pathogenicity, as the virulence factor of the bacteria increases when taken through ASR.

The Development and of Characterization of nanocomposites‘

D. Purvis, Dr. S. Ramakrishnan, Dr G. Louis Chakkalakal

FAMU Bridges to the Baccalaureate in the Biomedical Sciences

Florida A&M University

FAMU-FSU College of Engineering

Tallahassee, Florida

The research goal is to create a nano-composite that can be used in the development of light weight aircraft wings for the U.S. Air Force.  This can be achieved by understanding the properties of silica in a known polymer matrix of polystyrene.

Synthesis of 2-Methyl Indanone derivatives as potential antipsychotics

Eduardo Sanchez, Edward Ofori, and Seth Y. Ablordeppey

FAMU Bridges to the Baccalaureate in the Biomedical Sciences
Florida A&M University
College of Pharmacy and Pharmaceutical Sciences
Tallahassee, Florida 32307

Previous affinity binding data on 5-chloro-2-(2-(3,4-dihydroisoquinolin-2(1H)-yl)ethyl)-2,3-dihydro-1H-inden-1-one(10) on the most recent class of the serotonin receptors, 5HT7 receptors (5HT7R) shows that it has high affinity for this receptor which has been linked to cognitive function in the human brain. Researchers opine that agents which interact with this receptor may be useful in restoring cognitive deficit seen in Schizophrenics. The lead compound 10 had a stereo center and attempts to separate the isomers has failed since the alpha proton is highly acidic and isomerization is inevitable. To solve this problem we synthetically introduced a methyl group at the 2-position of the indanone to form 5-chloro-2-(2-(3,4-dihydroisoquinolin-2(1H)-yl)ethyl)-2-methyl-2,3-dihydro-1H-inden-1-one (18). This work is still in progress and we anticipate that the biological evaluation for this agent will be similar to the biological profile of compound 10.

Metagenomic Analysis of Soil Bacterial Diversity at a Geothermal Hot Spring, Manikaran, India

Marquise Cromartie1, Gregory Hitz2, Ashvini Chauhan2

1FAMU Bridges to the Baccalaureate in the Biomedical Sciences, Florida A&M University
2School of the Environment, Florida A&M University, Tallahassee, FL-32307

A metagenomic analysis was conducted on genetic information obtained from a soil-mousse sample at a geothermal spring in Manikaran, India. MG-RAST analysis identified bacteria as the dominate domain and species of Firmicutes as the most prolific bacteria. A heat map indicates that Firmicutes, Actinobacteria, and Proteobacteria are highly represented between samples.

Electroless Catalysts for the Degradation of Ibuprofen

Michelle Perez, Egwu E. Kalu, Shannon Anderson
TCC-FAMU Bridges Research Summer Internship
Department of Chemical & Biomedical Engineering
Florida A&M University, Tallahassee, FL 32310

The catalytic degradation of the common pharmaceutical, ibuprofen, was investigated using noble metal oxide catalysts prepared using electroloess deposition technique. The catalysts were synthesized by polymer-stabilized Pd nanoparticle-catalyzation and activation of Al2O3 or TiO2 substrate and electroless plating of the substrate for selected time lengths. Catalytic activity of synthesized catalysts was tested for the degradation of ibuprofen from an aqueous solution. Reaction samples were analyzed using UV-Visible spectroscopy. The effects of electroless catalyst composition and temperature on degradation rates will be analyzed and discussed.

Effects of Apigenin on hydrogen peroxide oxidative stress induced N2A cells

[Priscilla K. Medehue] 1, [Godwin Denyo] 2, [Dr. Tracy A. Womble, PhD] 3

FAMU Bridges to the Baccalaureate in the Biomedical Sciences
Florida A&M University
College of Pharmacy and Pharmaceutical Sciences
Tallahassee, Florida 32307

Hydrogen peroxide has been proven to induce oxidative stress on cells.1 Using this knowledge the concentration that will result in 60-70% viability was tested. Oxidative Stress is the imbalance between the production of free radicals and the ability of the body to counteract or detoxify their harmful effects through neutralization by antioxidants. Oxidative stress leads to many pathophysiological conditions in the body. Some of these include neurodegenerative diseases such as: Alzheimer’s, Parkinson’s, Huntington’s, Amyotrophic lateral sclerosis (ALS) and strokes. Nuero-2a cells from mice brains are neuronal like. Apigenin is  naturally occurring, abundantly present in common fruits and vegetables including parsley, onions, oranges, tea, chamomile, wheat sprouts and some seasonings. The processes that were performed was cell culturing, treatment of cells with H2O2 and Apigenin, these treatments were evaluated by Alomar blue viability assays. The hydrogen peroxide concentration was determined to be 50 uM and the Apigenin was not received so that was not carried out to receive results. In conclusion, we found that 50 uM of hydrogen peroxide was the lowest concentration that resulted in a 60-70% viability of the cells. Further studies with Apigenin are underway.

Rheology of Colloidal gels

  1. Benjamin, S. Ramakrishnan, PhD, H. Washington

FAMU Bridges to the Baccalaureate in the Biomedical Sciences
Florida A&M University
College of Pharmacy and Pharmaceutical Sciences
Tallahassee, Florida 32307

The purpose of this research was to study the flow and elasticity of colloidal suspensions, as well as the particle interactions that give rise to these flow properties; all to eventually be able to control these suspensions. While working in Dr. Ramakrishnan’s lab, I was able to use the Rheometer to observe the different flow properties of various commercial fluids. Of the fluids tested, 2 were Newtonian fluids, while the remainder were Non-Newtonian fluids. Although the research shown is minimal, the expectation is that eventually, through extensive studies, a connection between particle structure and fluid function will be made, making it possible to fully control these suspensions.

Synthesis and Cytotoxic Activity of Coumarin-based Benzopyranone Derivatives Containing Basic Amino Side Chain in Human Breast Cancer cell Line

Richard Torres a,   Musiliyu A. Musa b and Akintunde Gbadebo c

FAMU Bridges to the Baccalaureate in the Biomedical Sciences
Florida A&M University
College of Science and Technology
Tallahassee, Florida 32307

Estrogen stimulates the proliferation of tumor cells through the estrogen receptor (ER) via the induction of nucleic acid synthesis and activation of growth regulatory genes. Currently, antiestrogens are used to block this estrogen-like action on tumor cells by displaying an antagonist action at the estrogen receptor (ER). 1 There are two classes of non-steroidal antiestrogen compounds; triphenylethylenes (e.g. TAM, 1) and benzothiophenes (e.g. RAL, 2) with the basic side chain (Figure 1).2 The nature of this basic side chain and its orientation relative to the ligand backbone play an important role in the determination of the tissue-selective activity. 3

Most recently, our group has been interested in the coumarin (2H-l-benzopyran-2-one) ring system (3, Figure 1) as a core pharmacophore of potential therapeutic agents for the treatment of hormone-dependent breast cancer.4 Studies have shown that coumarin and derivatives demonstrate growth-inhibitory activity in human cancer cell lines, such as A549 (lung), ACHN (renal), H727 (lung), MCF-7 (breast) and HL-60 (leukemia), in addition to prostate cancer, malignant melanoma, and metastatic renal cell carcinoma.5 SP500263 (4, Figure 1), coumarin derivative containing basic side chain used in hormone-dependent breast cancer treatment, functions as potent antiestrogen in both in vitro and in vivo models of breast cancer and potently, inhibits estrogen-dependent MCF-7 proliferation with similar IC50 value to that of tamoxifen. 6