Darwin thinking

Poster Session

The Genetic Convergence of Carnivorous Plants

Authors: Moonia Ammari, Dr. Matthew L. Aardema

Abstract: The morphological traits associated with plant carnivory have independently evolved over several orders within the division of Angiosperms. Despite the lack of shared ancestry, carnivorous plants have similar adaptation to poor nutrient environments which suggest evolutionary mechanisms at play. This research aims to display the genetic underpinnings of trait convergence to each species of carnivorous plants to its non-carnivorous orthologues along with gene-related function. Here we use a computational approach to examine the genetic patterns and statistical significance of plant genes against Arabidopsis thaliana, to reflect plant carnivory. The Gene Ontology (GO) coding allows us to access the quantitative significance of functional genes across all taxon sequences. The gene functionally associated with methylammonium transmembrane transporter activity, more exclusively known as aquaporin, has been silenced in all carnivorous taxa while remaining present in the non-carnivorous taxa. In relation to other findings, the availability of genomic sequencing data enhances the ability to address statistical significance more accurately. The loss or Inactive mutations of this gene may have resulted at the expense of phenotypic expression of the trapping mechanism. This event allows us to determine how and why plant carnivory is determined on a genomic level in relation to non-carnivorous species.

How Can a Frozen Frog Maintain Functioning Muscles and Liver?

Authors: Nancy Alomawi, Dr. Kevin Bilyk.

Abstract: Freeze-tolerant frogs can survive freezing of up to 2/3 of their body water. During this time, they have no heartbeat and no brain activity. In species such as Cope’s Grey Treefrog they survive such freezing by accumulating cryoprotective compounds of hepatic origin including glycerol, urea, and glucose. As these cryoprotectants are produced in the liver we hypothesized that the expression of genes that were related to cryoprotectant mobilization and stress tolerance would be different across tissues in response to cold. To investigate we used RNA sequencing & gene expression analysis to compare the hepatic and muscle transcriptomes among frogs that were cold and frozen.

Comparison of Wetland Seeds

Authors: Martha Mahady, Dr. Dirk Vanderklein

Abstract: Wetlands support a diverse community of plant species.  The seeds of these plants are adapted to survive and germinate in wetland conditions.  Germination rate is affected by soil conditions, including temperature, light, oxygen, soil structure, and water. The size of seeds is an important characteristic, affecting seed dispersal, drought survival, and nutrition resources available to the seedling.  The mass of the seeds of four wetland plant species, purple loosestrife (Lythrum salicaria), winged loosestrife (Lythrum alatum). Joe Pye weed (Eupatorium macalatum), and common cattail (Typha latifolia), was measured and seeds of these species were sown in pots in the greenhouse to determine the germination rate.  The seeds of Joe Pye Weed were small with an average mass of 316.2 micrograms. However, those of the other species were extremely small, purple loosestrife from Sussex County 47.5 micrograms, purple loosestrife from United Kingdom 45.5 micrograms, winged loosestrife 14.4 micrograms, and common cattails 39.8 micrograms.  The germination rate of purple loosestrife seeds was significantly higher than that of the other species.  The high germination rate of purple loosestrife, which is invasive in North America, may be one factor in its greater competitiveness, enabling it to spread rapidly across the landscape in a new region. 

Parasite variability in the invasive crayfish, Faxonius rusticus, in Northern New Jersey

Authors: Nathan Klunk, Dr. Matthew Aardema

Abstract: The Northeast United States has very thoroughly been invaded by the well known invasive crayfish species Faxonius rusticus (rusty crayfish). Similar to the exploration of man to new regions, the introduction of invasive species can cause the introduction of new diseases and parasites while the invader has to deal with the already existing diseases and parasites. Most notable of these is the crayfish fungal plague, which has had very poorly understood distribution in the United States. In this study, I collected rusty crayfish in Northern New Jersey to clarify locations they exist in the state and dissected the collected individuals to look for various parasites or signs of parasitism as an insight into potential fields of question. My results suggest that rusty crayfish are more likely to express parasitism while in a cobble substrata compared to a clay/mud substrata. In addition, most observed signs were primary indicators of the fungal plague. Although rusty crayfish and the fungal plague are well understood separately, these findings should indicate that the two organisms need to be looked at more closely together to interrogate their impacts further and clarify how impactful the plague truly is to the rusty crayfish population.

The Potential Use of ICER Peptides in Cancer Treatment

Authors: Karim Homsi, Daniela Alves, Keith Lange, Angelo Cirinelli, Dr. Carlos Molina

Abstract:  Although chemotherapy, radiation, immunotherapy, and surgery can be effective cancer treatments, cancer research is always on a mission to find a more efficient, less expensive way to treat cancer. The use of peptides in cancer-related treatments is not as common as the treatments mentioned above, but is a very promising cancer research field. Cell-penetrating peptides have the ability to deliver anti-cancer therapeutics to cancer cells. Inducible cAMP Early Repressor (ICER) is a transcription factor that is found in all eukaryotes. Deregulation of ICER protein is a common phenomena in many cancers, including skin cancer melanomas. ICER possesses tumor suppressing abilities. Overexpression of ICER blocks cells in mitosis, eliciting cell death and therefore, halting the tumorigenicity of cancer cells.  Our data shows that ICER possesses cell penetrating peptide properties and is able to penetrate melanoma cells in culture. This cell-penetrating characteristic of ICER protein could potentially be used to eliminate tumor cells by apoptosis, setting the stage for the development of novel treatments for cancer. 

Binding Affinities of Selected Single-stranded DNA aptamers for the Detection of Ranavirus

Authors: Stephanie Zapata, Corbin Hudson, Maria Fahmy, Evelyn Visan, Dr. Kirsten Monsen, Dr. Nina M. Goodey, Dr. Lisa Hazard, Dr. Ueli Gubler

Abstract: The current molecular diagnostic methods for detection of the deadly pathogen Ranavirus are time-consuming, expensive, and require specialized training and equipment. When surveying a population for the presence of a pathogen, time is a valuable resource. Faster detection could have significant impacts on commercial value of organisms, species conservation, and potential containment of the pathogen. Ranavirus is a double stranded DNA virus that can cause mass mortality events in amphibians, reptiles and fish. The disease can have the potential to wipe out entire species if not controlled, having a 90-100% mortality rate. Recent methods of development for a rapid pathogen test have involved target detection probes using aptamers, or short single stranded DNA as molecular probes. Studies have shown that molecular probes are more stable and cost-efficient than antibodies, and are also easier to modify. Using an existing starting library of 1014 non-specific aptamers that have been previously generated via SELEX methods, and are specifically selected to bind to Ranavirus, the objective is to determine the binding affinities of the selected aptamers to the Ranavirus major capsid protein (MCP) target using electrophoretic mobility shift assays (EMSA). The results of this research will provide a better understanding of the aptamers high affinity and specificity for MCP, and the selected aptamers can therefore be used for the development of a rapid pathogen test that can be used in real-time in the field. 

Analyzing the effect of tubulin glutamylation on localization of the microtubule-associated protein RPIL-1 in C. elegans sensory neurons

Authors: Ema Ramicevic, Yandis Toyos, Kaylee Cuyuche, Dr. Robert O’Hagan

Abstract: Microtubules (MTs) are cytoskeletal fibers composed of polymers of tubulin. MTs control cell division, growth, motility, and the movement of vesicles from neuronal cell bodies to axons. The Tubulin Code hypothesis suggests that individual microtubules are composed of different tubulin isotypes that have been post-translationally modified to specialize their function. MAPs (Microtubule-Associated Proteins) are also thought to bind to MTs to regulate the MT cytoskeleton. The Tubulin Code and MAPs are involved in neurodegenerative disorders in humans, as regulation of MTs is essential for healthy neurons, because MTs develop and maintain axons and dendrites.  Retinitis pigmentosa (RP) and Occult Macular Dystrophy (OMD) are neurodegenerative disorders that ultimately result in blindness.  OMD can be caused by mutation of a gene encoding a doublecortin-family MAP called RP1-like1, which is heritable as an autosomal dominant disease and results in cone photoreceptor cell degeneration, despite the Fundus appearing normal. To investigate the function of RP1-like1, we identified a homolog in the nematode C. elegans by homology search.  We created transgenic animals expressing a recombinant RPIL-1::GFP transgene fusing the RPIL-1 protein to the green fluorescent protein to mark the expression of RPIL-1.  We find that RPIL-1::GFP is expressed throughout the somata, axons, and dendrites of ciliated sensory amphid and phasmid neurons, but excluded from sensory cilia.  We hypothesize that the Tubulin Code and MAPs interact. Therefore, we are analyzing the subcellular localization of RPIL-1::GFP when enzymes that add or remove glutamylation sign-posts on MTs are mutated. We hope to further understand how RPIL-1 is targeted to particular MT networks, and how its function affects MTs.

Targeting bacteria causing joint infections using modified EGCG derivatives: EGCG-S AND P-EGCG

Authors: Kavneet Chahil, Mohamad Gacham, Abraham Helwani and Dr. Lee H. Lee

Abstract: Prosthetic joint implantations have revolutionized orthopedic medicine by allowing individuals to regain physical function with minimal side effects. However, they are susceptible to post- surgical complications due to bacterial infections. Three significant bacteria that are responsible for the development of joint infections include Staphylococcus epidermidis, Staphylococcus aureus, and Pseudomonas aeruginosa; which act by adhering to the prosthesis and form biofilms thus becoming antibiotic-resistant. It has been reported that green tea polyphenols, which are extracted from the leaves of the Camellia sinensis plant, have antibacterial, antioxidant, and anti- inflammatory properties. The purified, and modified epigallocatechin-3-gallate-stearate (EGCG- S) and palmitoyl-epigallocatechin-3-gallate (P-EGCG) are more efficient and have shown their synergistic effect on antibiotics indicating they could potentially play a positive role in the elimination of infection. In this study, the inhibitory effect of EGCG-S and P-EGCG with or without two antibiotics (Bacitracin or Polymyxin B) on the three most prominent bacteria was investigated. Colony-forming unit (CFU) assays were used to determine the percent of inhibition by the tea polyphenols. Time course studies were performed using CF1 and CF2 formulations to determine the percent of inhibition. Congo Red assays were used to analyze the effect of tea polyphenols on biofilm. This study showed that EGCG-S and P-EGCG, when used in combination with Bacitracin or Polymyxin B resulted in the best antibacterial effect and could be used as a treatment against bacteria that cause joint infections.

Analysis of the efficiency of EGCG-Stearate on antibiotic resistant bacterial Acibetobacter baumannii

Authors: Raisa Da Silva, Dr. Lee H. Lee

Abstract: The inappropriate use of antibiotics throughout the years has led the world to face an emergence of multidrug resistant microorganisms that have become one of the top three threats to global public health. This has resulted in the necessity of discovering alternative drugs and methods to combat this problem. Acinetobacter baumannii is one of the six antibiotic-resistant opportunistic bacteria species responsible for most of the current nosocomial infections. The main goal of this research was to determine the efficiency of EGCG-Stearate (EGCG-S) used alone or in combination with erythromycin against the growth and biofilm formation of A. baumannii. Standard serial dilutions were performed to determine the optimal dilution factor for these bacteria. Congo Red Biofilm Assay was conducted to study the effectiveness of 500 ug/mL of EGCG-S and EGCG-S in combination with erythromycin against biofilm formation. EGCG-S presented 99.65% of inhibition and a 2.46 log reduction of A. baumannii when used by its own after one hour of treatment. In the Congo Red Biofilm Assay, EGCG-S showed to inhibit the formation of biofilm when used by its own and in tangent with the antibiotic erythromycin. These results indicate that EGCG-S is a potential therapeutic agent to be used as an alternative treatment against A. baumannii. 

In-vitro assessment of Methylsulfonylmethane cell regulation via MiRNA Profiling and Cell health assays

Authors: Charles Ezenwanne, Dr. Ann Marie DiLorenzo

Abstract: Methylsulfonylmethane has been used for years as a dietary supplement for numerous benefits, one of them being it’s Anti-Inflammatory and prevention of the over-stress of immune cells. Inflammation has been found to link to certain types of cancers such as colorectal cancer. MicroRNA are small Non-coding RNAs that play key roles in the regulation of gene expression. It fine-tunes the expression as much as 30%. It acts during the post transcription level. Studies have been discovered in using miRNA-based therapy as a way to combat cancer, due to miRNA’s role in the regulation of tumor suppressors. Studies also show MiRNA has a role in cancer prevention, because Specific miRNA is found to be upregulated and downregulated when cells go into metastasis. Using this information, I’m going to test specific miRNA and see if these same ones are upregulated and downregulated when treated with Methylsulfonylmethane. My goal is to further research Methylsulfonylmethane’s (DMSO2) cancer fighting properties via MiRNA profiling. This research is of value because miRNA is still currently studied and may lead to new innovation in cancer research in which cancer is still a growing issue globally. Methylsulfonylmethane has already proven to have anti-inflammatory and prevent over-stress of immune cells, we can further explore this beneficial supplement and its possible role in cancer therapy. The main question of this study is Can Methylsulfonylmethane play a role in cancer research?

Antibiotic effects of green tea polyphenol EGCG-S on nosocomial bacteria Staphylococcus aureus and Klebsiella pneumoniae

Authors: Calvin Taylor, Dr. Lee H. Lee

Abstract: The ESKAPE bacteria (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) have developed their multidrug resistance through adaptation to excessive drug use or prescription, unnecessary antimicrobial utilization and substandard pharmaceuticals. They are the major cause of nosocomial infections throughout the world. With the over growing concern of antibiotic resistance in treating bacterial infection, search for novel alternative therapy is essential. Green tea polyphenols in herbal plant Camellia sinensis have shown to have antibacterial activity. A modified and stable form of EGCG, epigallocatechin-3-gallate stearate (EGCG-S) further showed their ability to enhance the antibiotic activity. In this study, EGCG-S was used to study its inhibitory effect on two of the ESKAPE bacteria Staphylococcus Aureus and Klebsiella Pneumonia. Colony forming unit assay was used to determine the % of inhibition and log reduction of EGCG-S treatment. Congo red assay was used to study its effect on bacterial biofilm formation. The organic formulation F1 and F2 containing palmitoyl-epigallocatechin-3-gallate (P-EGCG) was also used to evaluate their antibacterial effect. Studies conducted showed that EGCG-S, have a positive effect on the inhibition of these bacteria and their biofilm. The formulations can efficiently inhibit the growth of the bacteria. This study suggests that EGCG-S is a valuable agent against antibiotic resistant bacteria.

Self-Conscious Emotions and the Right Fronto-Temporal and Right Temporal Parietal Junction.

Authors: Adriana LaVarco, Nathira Ahmad, Qiana Archer, Matthew Pardillo, Ray Nunez Castaneda, Anthony Minervini, and Julian Paul Keenan.

Abstract: For more than two decades, research focusing on both clinical and non-clinical populations has suggested a key role for specific regions in the regulation of self-conscious emotions. It is speculated that both the expression and the interpretation of self-conscious emotions are critical in humans for action planning and response, communication, learning, parenting, and most social encounters. Empathy, Guilt, Jealousy, Shame, and Pride are all categorized as self-conscious emotions, all of which are crucial components to one’s sense of self. There has been an abundance of evidence pointing to the right Fronto-Temporal involvement in the integration of cognitive processes underlying the expression of these emotions. Numerous regions within the right hemisphere have been identified including the right temporal parietal junction (rTPJ), the orbitofrontal cortex (OFC), and the inferior parietal lobule (IPL). In this review, we aim to investigate patient cases, in addition to clinical and non-clinical studies. We also aim to highlight these specific brain regions pivotal to the right hemispheric dominance observed in the neural correlates of such self-conscious emotions and provide the potential role that self-conscious emotions play in evolution.

Metal Chelators Ethylenediaminetetraacetic acid (EDTA) and N,N,N′,N′-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN) Interfere with Carbonic Anhydrase (CA) Activity in Wilbur-Anderson Assay (WAA)

Authors: Madeline Lucas*, Vladislav Snitsarev (*Supported by LSAMP)

Abstract: In our effort to set up a laboratory for undergraduate students with the tentative title “Studying enzymatic activity of Carbonic Anhydrase”, we successfully determined carbonic anhydrase activity and demonstrated its inhibition by Acetazolamide, a potent CA inhibitor, using the Wilbur-Anderson assay (WAA). WAA hinges upon determining the time T (in seconds) required for 4 mL of saturated with carbon dioxide (CO2) water to lower pH from 8.3 to 6.3 of the mixture when added to 6 mL of 0.02M Tris buffer at 0°C without (T0) and with enzyme (T). Activity (A) is defined as A = 2*(T0/T-1) Wilbur-Anderson Units (WAU). In n = 16 blank trials, T0 = 75.0±10.7s (Mean±Standard deviation), and with 0.001mg of CA in n = 12 trials, T = 50.2±8.1s, A = 0.99±0.66 WAU (p < 0.001). When we used EDTA, a carboxylic high-affinity metal ion chelator, to inactivate CA, a Zn2+-containing enzyme, we unexpectedly encountered that in the presence of the sodium salt of EDTA (Na2EDTA): 1) T0 was significantly slowed in a concentration-dependent manner: for 5μM, 50μM, and 500μM, T was 84.3±7.9s (n = 10, p < 0.05), 108.5±15.6s (n = 7, p< 0.001), and 130.8±22.7s (n = 10, p < 0.001), respectively; and 2) unexpectedly in the context of the previous finding, Na2EDTA (5μM) significantly enhanced CA activity (T = 38.5±4.0s, A = 1.84 WAU, n = 10, p < 0.001) that was abrogated by Acetazolamide (10μM), a potent CA inhibitor (T =138.3±10.4s, n = 7, p < 0.001). TPEN, a pyridinic non-carboxylic metal ion chelator that is more potent in metal ion chelation that EDTA, significantly slowed T0 (99.5±15.9s, n = 4) similar to Na2EDTA, and significantly enhance CA activity (TE = 41.1±3.8, n = 4, p < 0.05) with equal potency. This indicates that chemically different metal ion chelators EDTA and TPEN interfere with the determination of CA activity in the context of WAA. Studies to explain these phenomena are underway with the hypotheses that the contaminating metal ions that catalyze CO2 hydration/dehydration without CA are responsible for the observed phenomena.