2017 Eastern New England Biology Conference
Abstracts
All are ordered by the last name of the first author

The effect of benzyl butyl phthalate exposure and isolation on Madagascar hissing cockroach behavior and response to visual and auditory stimuli
Akorfa Adobor, Zahra Al-Alawi, and Lisa A.E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
This pilot study was conducted to determine the effect of benzyl butyl phthalate (BBP), a known plasticizer and suspected endocrine disrupter, on female Madagascar hissing cockroaches. Adult female cockroaches were assigned to one of three groups: isolated / untreated (N=10), isolated / dosed (N=10), and communal untreated (N=10). Changes in total food consumption, leg movement, and response to both auditory and visual stimuli did not differ significantly among any of the groups. Behavioral responses (agitation and lethargy) did not differ significantly between the isolated untreated and isolated dosed groups, but communal untreated females displayed higher levels of agitation than both isolated untreated females (p = 0.0152) and BBP-dosed females (p = 0.0168). Since the visual and auditory stressors chosen for this experiment did not evoke measurable differences in behavior, they are not usable tests to determine BBP-induced behavior changes. The data suggests that specific behavioral differences do occur in response to environmental change. In this pilot study, however, isolation of the social insects played a larger role than exposure to BBP.
 
 
Discovery of regulators of the master regulator of the general stress response, RpoS
Wajdan Almajed, Mohammed Aloufi, Iyonna Couvertier, Meisha Dorlean, Marissa Freely, Sharon Goh, Marcellous Govind, Olivia Huber, Fatimah Jalih, Jade Kaplan, Marissa Kaufman, Kaylee Lampert, Shaza Mass, Imtithal Mutaen, Dylan Parsons, Joseph Pezzolesi, Sara Trimble, Christopher Williams, Alice Wynn, Thomas Tran, Arup Dey, and Celeste Peterson        
Suffolk University
Poster Session B: 4:00-5:15
 
Most bacteria have developed certain ways to overcome environmental threats, such as entering into a stationary phase. RpoS, a master regulator of stationary phase, controls which prokaryotic genes are transcribed when the bacterial cells are dormant. However, little is known about the genes that control RpoS itself or the mechanism by which these genes express/inhibit RpoS. In this experiment, we created a plasmid library of the E. coli genome to discover novel genes that regulate an RpoS’-‘LacZ fusion expression and performed assays to determine if the regulation of RpoS is at the level of transcription or post-transcription. We identified five genomic regions that upregulated RpoS and four genomic regions that downregulated RpoS. While some of the genomic regions contained full protein coding genes (such as rbsD, yfaS, yfaT hua , nadA ) including many with unknown functions, other regions contained tRNAs or regions with potential sRNAs (such as the intergenic regions between adiY and adiA) . Only one of the nine regions contained a gene previously identified to regulate RpoS ( huA) . Using betagalactosidase assay with different lacZ fusions, we determined that eight of the nine regions were regulating RpoS at the level of translation and/or protein stability rather than transcription. Thus, we have identified candidate novel regions that may regulate post-transcription expression of RpoS.
 
 
Synergistic Antibacterial Activity of Turmeric, Ginger, Honey and Garlic Extracts on Selected Probiotic and Pathogenic Bacteria
Faith Bertrand, David Luzgin, and Rekha Singh
American International College
Poster Session B: 4:00-5:15
 
Turmeric is a spice from the root Curcuma longa, a member of the ginger family. Curcumin, a biological ingredient in turmeric has been shown to have anti-inflammatory, anticancer, and antimicrobial properties. In this study we analyze the antibacterial properties of turmeric alone and in combination with ginger, honey and garlic extracts. Many studies have investigated antibacterial effects of these individual extracts but not their different concentration blends to identify their enhanced efficacy, if any. The impact of the above extracts and blends on the growth of selected probiotic bacteria and pathogenic bacteria would be assessed in this study. This research could provide a basis for further exploration of the possible benefits of the above extracts as effective natural antibacterial agent.
 
 
Influence of predator-escape behaviors and habitat on hindlimb morphology      
Abdalaziz Biyari and Eric W. Dewar
Suffolk University
3:00–SMT 317
 
Animals change their escape behaviors largely depending on their habitat when approached by a predator. Living artiodactyls are morphologically diverse in their hindlimb skeleton partly because they specialize in different predator-escape behaviors. To examine this morphological variation, we generated three-dimensional models of skeletal morphology using photogrammetry. The aim of this study was to analyze landmark-based shape variation using 3D geometric morphometrics to determine the influence of predator-escape behaviors on the morphology of the hindlimb skeleton in both living and extinct artiodactyls. Angle of escape, duration of trotting and stotting, and change in elevation behaviors during flight were introduced here. We wanted to seek morphological convergences among these different predator-escape behaviors and habitats influence to broaden our understanding in extinct species habitats and lifestyles. It was found that when the approacher was close during initiation of pursuit, deer fled relatively shorter distances with more acute escape angles forcing predators to change direction suddenly, which hinders their speed. When the threat of an encounter increased, deer tended to flee uphill and into taller vegetation, and trotting and/or stotting behaviors were observed.
 
 
Comparison of the use of 16s rRNA DNA sequencing and the Biolog microbial identification system for the identification of unknown microbes collected by students in a pathogenic microbiology class
William Bomentre and C. Eggers
Quinnipiac University
Poster Session A: 1:00-2:15
 
            In the pathogenic microbiology course at Quinnipiac University, students identify unknown bacteria recovered from their oropharynx or skin, or the stool of domestic animals; these identifications are largely based upon a limited number of biochemical tests and sequencing of a portion of the 16S ribosomal RNA gene. In a teaching laboratory, technical difficulties and the limited nature of the data acquired suggested the need for an additional identification technique. The purpose of this research was to evaluate the ease of use and accuracy of identification by the Gen III Microbial Identification System (Gen III MIS; Biolog, Inc.) relative to sequencing.  The Gen III MIS uses a 96-well format to test the metabolic activity and sensitivity of an organism utilizing a variety of compounds; the resulting pattern is compared to a database to determine a likely identification.  To test the concordance between the two identification systems, 20 random bacterial samples were resurrected from freezer stocks.  After determining Gram stain, DNA was extracted from cells and the 16S rRNA was amplified and sequenced. The cells were also inoculated onto a Gen III MicroPlate.  We were able to amplify the 16S rRNA gene and identify database matches from 16 of the 20 samples.  The results from the Gen III MIS for fourteen of these sixteen samples were consistent with the sequences and results of the Gram stain; the other two matched the Gram stain results, but not the sequence. Of the four remaining samples from which the 16S rRNA gene was not amplified, three were able to be identified using the Gen III MIS. One sample was not identified by either method. Based on these results, we find that the Gen III MIS is an effective method for the identification of microbial unknowns, with a reliability similar or better than 16S rRNA sequencing.
 
 
Impact of BBP and acetone exposure on Betta splendens health, behavior, and neurotransmitter production   
Makenna Camara, Jordyn Platz, and Lisa A.E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
The objective of this study was to determine the impact of acetone and benzyl butyl phthalate (BBP) on health, behavior, and neurotransmitter production in male Betta splendens. Each fish (N=22) was assigned to one of four groups (control, 0.1 ppm acetone vehicle, 0.1 ppm BBP, or test group). The mean somatic index (SI) was calculated for each group at the beginning and end of the 5.5 week exposure period. While SI decreased in control and BBP groups (11.61% and 20.05%, respectively), it increased 11.84% in the acetone group. The greatest overall change in SI, however, occurred in the BBP group (20.05%). Control, acetone, and BBP fish responses (passive, moderate aggressive, and aggressive) to test fish were also assessed. Passive behaviors increased in all treatment groups, but the greatest change was noted in the control group (156.4% increase). A 100% decrease in moderate aggressive behavior was observed in control and BBP groups but not among the acetone group (71.43%). Similar results were observed for aggressive behaviors; it should be noted that fish in the acetone group failed to exhibit aggressive behaviors in either pre- or post- exposure assessments. While no significant differences in brain dopamine was observed among any of the groups, significant increases in the production of serotonin (141.84%) were observed in the BBP group compared to its control counterpart. In summary, BBP and acetone alter passive behaviors in male Betta splendens, but they differ in their impact on health, moderate aggressive, and aggressive behaviors.
 
 
Investigating a novel secondary mitotic exit function for Lte1 in S. cerevisiae     
Andreia Carvalho and Anupama Seshan
Emmanuel College     
Poster Session A: 1:00-2:15
 
The Mitotic Exit Network (MEN) is a signaling pathway that acts in late anaphase of mitosis. The purpose of MEN is to allow cells to progress from late anaphase into G1. Occasionally, the mitotic spindle mispositions such that anaphase occurs entirely within the mother cell. In such cases, inhibition of the MEN by the spindle position checkpoint (SPoC) is essential to prevent aneuploidy. The spatial segregation of Lte1, a MEN activator, and Kin4, and MEN inhibiting kinase, is central to SPoC function. Lte1 binds to and inhibits Kin4. We hypothesize that Lte1 has an unknown secondary function in the MEN. To test this hypothesis, we used a mutant allele of this protein, named lte1ΔEcoRI, which is missing the Kin4-binding domain. With it, we created plasmids containing lte1ΔEcoRI-GFP fusions and transformed these into E. coli, in order to obtain defective and hyperactive alleles of this protein. GFP-positive colonies were obtained, but the plasmids could not be sequenced due to the low DNA yield. To continue investigating the putative secondary function of Lte1, we analyzed whether strains containing lte1ΔEcoRI could bypass the SPoC. When comparing the experimental strain containing lte1ΔEcoRI to the negative controls, we found that this mutant allele bypasses the SPoC and allows cells to exit Mitosis inappropriately. The discover highlights that Lte1 has a Kin4-independent function in activation of the MEN and that lte1ΔEcoRI may be a hyperactive allele of Lte1.
 
 
From Zinc Fingers to Cpf1: Editing the Genome
Lindsay Cathcart
Emmanuel College
Poster Session A: 1:00-2:15
 
Gene editing is the alteration of a specific region of the genome through the insertion or deletion of nucleotides facilitated by site-specific nucleases. Genome editing is analogous to the “find and replace” feature in a word processor; the site-specific nucleases can “find” a specific region of genomic DNA and effectively edit, or replace, the bases in that location.  Zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENS), CRISPR associated protein 9 (Cas9), and centromere and promotor factor 1 (Cpf1) are site specific nucleases that can edit many species’ genomes.  In addition to these enzymes, more complex “fusion enzymes” have been engineered to edit single bases of the genome at a specific location.  Of these enzymes, Cas9 has garnered the most attention as it is extremely efficient and user friendly.  Initially described as a bacterial adaptive immune system, the CRISPR-Cas9 system can be applied with relative ease to create site specific DNA edits.  Cas9 requires a RNA molecule, termed guide RNA, to provide the specificity to the genomic region to be edited. With these enzymes, edits can be made to inactivate gene expression via non-homologous end joining (NHEJ) mediated nucleotide insertions or deletions, or facilitate the repair of a mutated gene or insertion of a gene through homology directed repair (HDR).  ZFN and TALEN mediated gene editing therapies are currently in the clinic, treating illnesses such as HIV and leukemia, whereas the first clinical trial utilizing CRISPR gene editing technology was approved during the summer of 2016.  Here, I will present a comprehensive review of the current state of the gene editing field.  The nuances of each enzyme type will be described, with emphasis placed on substrate requirements and mechanisms.  Lastly, I will discuss the applications of gene editing technology to create next generation therapeutics for genetic diseases and cell therapy.     
 
 
Laboratory evolution of probiotic strains with superior effectiveness against the tooth decay- causative agent Streptococcus mutans
Kendall Chase and Jessica A. Smith
American International College
Poster Session B: 4:00-5:15

Taking care of your mouth, teeth and gums is a worthy goal in and of itself. Good oral and dental hygiene helps prevent bad breath, tooth decay, gum disease, and can help you keep your teeth as you get older. Dental decay is primarily caused by fermentative, acid-producing bacteria that reside in the mouth, such as Streptococcus mutans. Probiotics have gained much attention lately for their ability to fight off disease causing pathogens. This study involves evolving probiotics isolated from both yogurt and tablets to become more effective in killing off S. mutans. Several probiotic lactobacilli have been isolated, and disks containing the lactobacilli strain were placed onto a plate containing S. mutans. A zone of clearing was observed from two of the probiotic strains, indicating these strains were able to release antimicrobial substances that inhibited the growth of S. mutans. Currently we are continuously co-culturing these lactobacilli strains with S. mutans in order to select for probiotics that evolve enhanced mechanisms for killing S. mutans. Once these strains have evolved we will identify the antimicrobial compounds and mechanisms used for toxicity against S. mutans. This research is beneficial to the dental community in developing novel probiotic strains that may be useful to individuals who are cavity-prone. In addition, this research will allow us to identify the evolutionary processes involved in probiotics and the mechanisms they use to kill off competitor pathogens.
 
 
Estrogen Receptor (ERα) expression influences behavioral plasticity and matching behavior of guppies (Poecilia reticulata)  
James Cheney and Bronwyn H. Bleakley
Stonehill College        
2:30–SMT 316
 
In a constantly changing environment, it is necessary for organisms to be able to change their behavior to increase their likelihood of survival and reproduction. The behavior of a social partner is an environment and interactions between social partners can influence the phenotype of a focal animal. Such “interacting phenotypes” are partially the result of indirect genetic effects (IGEs), where the genes of one partner influences the phenotype of another. IGEs influence the antipredator behavior of guppies, Poecilia reticulata, but little is known about the physiological mechanisms that underlie such behavioral plasticity in response to social partners. This experiment investigates the influence of the estrogen steroid hormone receptor (ERα) on the plasticity of the behavior exhibited by a wildtype guppy that arises partially from the indirect genetic effects of two different strains of social partners. A female wildtype focal fish and was paired with a male and female guppy from two strains with noted behavioral differences and placed in a tank with a model of a predator. The shoaling, orientation, inspection, and proximity behaviors of the guppies were recorded and and the expression of the estrogen receptor α (ERα) was measured using quantitative PCR. The results showed a significant relationship between the expression of ERα and the guppy’s ability to match the shoaling behavior of its partners. There was no significant relationship between the level of ERα expressed and the behavioral plasticity of the wildtype guppies. The ability of a guppy to match the behavior of its social partner and its behavioral plasticity were only slightly correlated. However, principle components analysis provides evidence for more complex relationships between the components of guppy antipredator behavior. These results started to reveal the possible mechanisms underlying the behavioral plasticity displayed by the guppies in an antipredator environment.
 
 
Pilot Study to Explore MSSA and MRSA Nasal Carriage by College Students
Geena Chiumento1, V. Bennett1, J. Richards1, M. O’Seaghdha2, S. Duty1, and E. Scott1
1Simmons College; 2Suffolk University
3:15–SMT 315
 
Staphylococcus aureus is an opportunistic bacterial pathogen that is prevalent in both nosocomial and community settings; it asymptomatically colonizes the nares of certain individuals but is capable of causing serious skin and soft tissue infections. Colonization rates of the nares of individuals is 25-33% for methicillin-sensitive S. aureus (MSSA) and 7-9% for methicillin resistant S. aureus (MRSA). Several predictors of S. aureus carriage have been reported, including clinical hospital experience, use of hormonal contraceptives and frequent visitation to the gym.
The aims of this study are to determine the carriage of MSSA and MRSA among a healthy population of college students, to identify possible correlates for carriage (determined through a lifestyle survey), and to analyze genetic commonalities between isolates. In the past two years, a total of 302 nursing or non-nursing students were recruited and consented to provide 2 nasal swabs, one for each nostril. Swabs were processed and determined to be S. aureus using standardized identification methods. Furthermore, positive S. aureus isolates were classified as being either MSSA or MRSA by evaluation of growth on media containing 8 μg/ml cefoxitin. All S. aureus isolates were subjected to PCR amplification and sequencing of the spa gene, to determine species relatedness. The MSSA carriage rate among this college student population, 23.84% (72/302), is consistent with the reported national rates in the US; however, the MRSA carriage rate, 0.33% (1/302), is lower than previous reports. This may be due in part to the stringent confirmation test employed for MRSA in this study. Significant and trending predictors of colonization include recent clinical nursing rotations, frequent gym use, and recent upper respiratory tract infections.
 
 
Isolation and characterization of biofilm-forming Pseudomonas syringae strain CC2 from medicinal Callisia fragans plant
Claudia Conde and Jessica A Smith
American International College
Poster Session A: 1:00-2:15
 
Callisia fragrans, commonly called “Golden mustache”, is a plant widely known for its antimicrobial, wound healing, antitumor and analgesic properties. Although the leaves of this plant are studied for their many medicinal properties, few research studies have focused on the phyllosphere of the leaf. In this study we enriched for and isolated microbial strains from C. fragrans leaves. Very few viable bacteria were enriched from the leaves, but one gram-negative coccobacillus species, CC2, was isolated. CC2 is a motile, biofilm forming, and fermentative organism. Sequencing of the 16S rRNA gene showed that CC2 is closely related to strains of Pseudomonas syringae. Further studies are currently underway to determine the type of symbiotic relationship between CC2 and C. fragrans.
 
 
Monitoring Cytotoxic Effects of Theurapeutic Cancer drug administration via Immunohistochemistry Staining             
Iyonna Couvertier, Eugen Dhimolea, Pallari Awaite, Weng Xiang, and Constatine Mitsiades
Suffolk University
Poster Session A: 1:00-2:15
 
        Therapeutic chemotherapy drugs are known to induce cytotoxicity in both non-cancerous cells and in malignant cells. Breast cancer cells cultured from patient organoids in collagen gel in vitro generated from the MCF7 cell line have stained positive with Estrogen Receptor α monoclonal antibodies against the known cell proliferation marker protein for Ki-67. Ki-67 protein is specific in its expression in the nuclei of cells. An average of 34.8% of cells tested positive for ERα monoclonal antibody sensitivity which directly correlates to the exhibition of the Ki-67 marker protein found in high expression in breast cancer; the standard deviation for breast cancer cells exhibiting ERα monoclonal antibody sensitivity was 22.39%. Based on the MCF7 cell lines proven sensitivity to ERα monoclonal antibody the chemotherapy drug Docetaxel was administered which induced cytotoxic effects in 80% of malignant breast cancer cells and 19% in non-malignant cells that presented with ERα monoclonal antibody sensitivity, the standard deviation for cytotoxic effects in ERα antibody sensitive breast cancer cells was 26.5%. Prostate cancer cells grown in vivo from patient derived cells, in mice generated from the MDA-MB-231 cell line were also treated with the chemotherapeutic drug Docetaxel which exhibited Androgen Receptor α monoclonal antibody sensitivity.
           An average of 28.3% of prostate cancer cells exhibited ARα monoclonal antibody sensitivity, and 68% of prostate cancer cells displayed cytotoxic effects after administration of Docetaxel with a standard deviation of 34.3%. It was concluded that Docetaxel administration best targets breast cancer cells that exhibit ERα monoclonal antibody sensitivity in combination with Ki-67 presentation. Docetaxel was shown to induce cytotoxic effects in both in prostate cancer cells and non-malignant cells to greater degree than what was shown in breast cancer cells.
 
 
Does the size and shape of the orbit distinguish nocturnality and feeding behavior in extinct carnivores?
Bailey Mae Damron1, Enian Kallamata2, Magen M. Hartnett2, Alyssa Montecalvo1, and Eric W. Dewar1 
1Suffolk University; 2New England College of Optometry     
3:15–SMT 317
 
Living carnivorans are diverse in skull morphology, daily activity period, and feeding style. Because predation, particularly nocturnal hunting, is highly reliant on vision, we wanted to know if carnivoran skull morphology—particularly the morphology of the orbit—were related to daily (diel) activity pattern, as it is known to be in primates. We photographed the skulls of museum specimens of 48 living carnivore species (n = 225) and three Eocene-age extinct species (n = 8). From this sample, we compared (1) the relative size of orbit and cross-sectional area of the optic canal, (2) the shape of the orbit using landmarks placed along the orbital rim, and (3) the overall proportions of the face, orbital region, and cranium. We compared these observations with categories based on evolutionary relationships, diel activity pattern, and bite force or feeding style.
The relative size of both the orbit and optic canal showed significant family-level differences, mainly separating mongooses and seals and sea lions from other groups (F10,25 = 5.20, P = 0.0004). Diurnal species had relatively larger optic canals than nocturnal, crepuscular, and aquatic species (F3,32 = 8.56, P = 0.0003). There was not a significant relationship between feeding style categories and the size of the optic canal.
We performed a generalized Procrustes analysis to compare the shape of the orbit in our sample. This landmark-based analysis showed that living felids had less rounded, more inclined orbital rims while canids were more rounded and had larger gaps between the zygomatic arch and frontal bone. Bears had less rounded, more medially-indented, and still more open orbital margins. Larger species were able to produce higher bite forces than smaller species, but the shape of the orbit did not change in tandem with skull length.
            Differences in skull proportion were noticeable at the family level. Among our Eocene carnivorans, we found that the small canid Hesperocyon overlapped with both canids and mongooses, while the Eocene felids Hoplophoneus and Dinictis demonstrated the early establishment of the typical “cat-like” skull proportions.
 
 
In vivo Antibody Affinity Maturation Without Detectable Initial BCR-Antigen Recognition
Colby Devereaux1,2, J. Silver, J2, T. Zuo2, P. Tong2, N. Chaudry2, A. De Paiva Granato2, R. Patel2, Y. Chen2, D.R. Wesemann2,3     
1Emmanuel College; 2Brigham and Women's Hospital; 3Harvard Medical School
Poster Session B: 4:00-5:15
 
There are two diversification mechanisms that generate protective immunoglobulin (Ig) responses to vast array pathogenic threats. Primary Ig diversification is a result of Ig Heavy (IgH) and Ig Light (IgL) chain variable region exon assembly from V, (D), and J gene segments during early B cell development. This primary Ig repertoire is expressed as IgM and IgD B cell Receptors (BCRs) on naïve B cells. Secondary diversification results from somatic hypermutation (SHM) of the Variable region exon, followed by selection in germinal centers (GCs), resulting in the generation of high affinity antibodies toward antigen. This collective process is called affinity maturation. Secondary diversification results from activation dependent somatic hypermutation. Current thinking holds that pathogen recognition by BCRs is required to initiate affinity maturation to produce higher affinity antibodies. However, the degree of recognition required by the primary repertoire has not been well defined. We hypothesized that activation induced SHM and selection in GCs could produce high affinity antibodies despite the lack of initial BCR recognition. To address this we examined antibody evolution utilizing a model system wherein the initial antibodies produced are unable to recognize the antigen (Ovalbumin). The murine model used is our HRO mouse, ‘H’ – HA-restricted BCR, ‘R’ – RAG1 deficient, O – Ova expressing T cells, and another genetic lesion (RAG k/o that disables further Ig gene assembly). HRO mice chronically immunized with Ova were able to eventually produce high affinity antibodies toward Ova. Development of anti-Ova antibodies was delayed and highly stochastic compared to wild-type controls. Further experiments showed that these mice were also capable of generating serologic response to Ova-Hapten conjugates, as well. These results indicate that activation mediated hypermutation harbors sufficient intrinsic ability to initiate antibody evolution without initial BCR engagement and recognition, often utilizing similar pathogenic binding regions (epitopes).
 
 
Production of Wheatgrass (Triticum aestivum) and Hybrid Striped Bass (Morone chrysops x M. saxatilis) using Aquaponic Gravel and Float Sub-Systems
Nicholas A. Dibble and Travis K. Yandow
Colby Sawyer College
2:30–SMT 317
 
Wheatgrass (Triticum aestivum) is a valuable monocot that has health benefits, based on its antioxidant levels, and economic demand. The objective of this study was to evaluate the growth, production and sugar content levels of wheatgrass grown in two different aquaponic sub-systems. Gravel bed and floating tray techniques were tested by measuring growth rate, wet weight, dry weight and percent brix of wheatgrass. Gravel was the most productive sub-system with a mean wet weight of 577.13 grams total, compared to a mean wet weight of 194.58 grams for float technique. However, the float sub-system produced a greater mean percent brix at 4.5%, which was 0.6% higher than the gravel beds. Based on these results, we recommend the use of a gravel bed system, provided that the wheatgrass is harvested at an earlier stage of development to increase mean percent brix levels, which decrease as the plants mature. As a quantitative measurement of the hybrid striped bass (Morone chrysops x M. saxatilis) production, we calculated the feed conversion ratio (FCR) of 1.42 and specific growth rate (SGR) of 1.19% per day. Additionally, the same methods were used to compare the wheatgrass variables between aquaponic, hydroponics (gravel sub-system) and soil growing techniques. We found that the hydroponics was the most productive at the scale utilized. Again, we recommend harvesting the wheatgrass sooner in development to consistently have a greater mean percent brix. Our overall objective was to develop a greater understanding of aquaponic sub-systems, and how to efficiently produce the greatest yield for small-scale sustainable operations.
 
 
Characterization of Putative Superinfection Immunity (SIE) Proteins from a Mycobacterium smegmatis Bacteriophage, Hopey
Brittany Doherty, Emilee MacLean, Reuben Sands, Janine LeBlanc-Straceski
Merrimack College
Poster Session B: 4:00-5:15
 
Lysogens, bacterial cells that contain a dormant copy of a virus integrated into the host genome, are protected from secondary infections by different virally encoded mechanisms.  Super Infection Exclusion  (SIE) can be achieved through releasing enzymes that alter a portion of a surface receptor thereby blocking entry of another incoming phage.  Exclusion proteins can also target and block the ejection and/or DNA delivery of other phages.  This project aims to characterize one of these mechanisms to identify the putative SIE proteins produced by the prophage that could act at the membrane’s surface to exclude or block secondary infection.  We selected the putative superinfection SIE exclusion protein genes from Hopey, a virus that infects Mycobacterium smegmatis, cloned them into a protein expression vector and, expressed this them protein in E. coli.   in large quantitiesThese proteins are being and purified for crystallization to be later crystallizedand X-ray diffraction to determine their protein structure.
 
 
Investigation of antimicrobial activities of host defense peptides 
Deepesh Duwadi1, Anishma Shrestha1, James Jukosky1, and Steven N. Fiering2
1Colby-Sawyer College; 2Geisel School of Medicine
2:30–SMT 315
 
Host defense peptides are small, cationic, and amphipathic molecules that kill a wide range of pathogens. They belong to the innate immune system in both vertebrates and invertebrates. Due to their low toxicity to mammalian cells and a broad antimicrobial activity, there has been increasing attention to their therapeutic usage. Previous research demonstrated that four peptides—DAN1, DAN2, HOLO1 and LOUDEF1—derived from recently sequenced arthropod genomes based on homology searches showed potent antimicrobial effects (James et. al, unpublished). In this research, we showed that 20 mg/kg of DAN2 protected E. coli infected mice from lethality, ensuring 100% survivability. Other concentrations of DAN2 --10mg/kg and 5 mg/kg-- showed 83.30% and 66.60% survivability in mice respectively. DAN2, when administered at 5 mg/kg, 10 mg/kg and 20 mg/kg reduced bacterial counts after 6 hours of bacterial infections in blood and peritoneal fluid of mice. We determined using flow cytometry that DAN2 compromises the integrity of the membrane. This study shows that our peptides can contribute to the development of novel antimicrobial agents.
 
 
An Analysis of Growth and Phytoremedic Abilities of the Invasive Plant Species Fallopia japonica and Alliaria petiolata in Arsenic-Polluted Soil
Erica Eustis
Simmons College
3:30–SMT 316
 
When studying invasive plants, urban environments are of particular interest because they have been proven to be at high risk for invasion from non-native plants. This study investigated interactions between invasive plants growing in urban environments and chemical pollutants found in urban soils. It was hypothesized that invasive plants are capable of growing and surviving in soil contaminated with high levels of arsenic, a common soil pollutant. Furthermore, based on existing evidence that some invasive plant species are capable of performing phytoremediation of soil pollutants, it was hypothesized that invasive plant species are capable of performing phytoremediation of arsenic. The invasive plant species Fallopia japonica and Alliaria petiolata were grown independently in soil treatments contaminated with varying levels of sodium arsenate heptahydrate. Growth abilities were assessed by measuring survival rates, leaf numbers, and plant masses. Assessment of phytoremedic abilities was performed by measuring levels of arsenic absorbed by plant tissues using X-Ray Fluorescence analysis. Soil samplings from Fallopia japonica and Alliaria petiolata populations found along the Muddy River in Boston were collected and subjected to X-Ray Fluorescence analysis in order to measure arsenic levels in natural invasive plant urban habitats. Successful growth of plants in contaminated treatments will suggest tolerance to soil pollution as a possible mechanism by which invasive plants outcompete native plants in urban environments. Absorption of arsenic by plant tissues will add to the growing body of literature that suggests invasive plants are capable of having a positive impact on the environment through the process of phytoremediation.
 
 
Towards the development of a CURE (classroom undergraduate research experience) for tuberculosis
Andrew Farinha and Padraig Deighan
Emmanuel College     
Poster Session B: 4:00-5:15
 
Bacteriophage (phage) are the most abundant biological entity on earth. It is estimated that there are 1031 phage that occupy every ecosystem in which their host bacteria reside. Perhaps not surprisingly, the genetic diversity within the phage population is enormous and most phage genes encode proteins with no known functions. After infecting a bacteria the phage life cycle may proceed in either a lytic or lysogenic life cycle. However, in both cases a proportion of the phage genes will ultimately disable essential host functions leading to host cell death. In this project we have taken the initial steps in the development of a classroom undergraduate research experiences (CUREs) that focuses on mining phage for proteins that deliver a lethal blow to their hosts. In particular, we are focusing on phages that infect Mycobacterium smegmatis, given that the largest collection of sequenced phages is for this organism, as determined by the SEA- PHAGES undergraduate program. We have optimized protocols where genes from any sequenced phage can be assembled into a shuttle vector, which allows for tight and controlled induction of the phage gene in M. smegmatis. Further, we have cloned over 30 genes from one phage and uncovered several genes that are lethal when expressed in M. smegmatis. We next plan to use a bacterial two-hybrid assay to uncover the protein partners for some of these lethal phage proteins. We hope that freshmen taking this CURE will 1. become inspired and confident about scientific research and will pursue studies in a STEM field, and 2. contribute to advances in uncovering novel antibiotic targets in M. tuberculosis, a disease-causing cousin of M. smegmatis, which infects one-third of the world’s population and is increasingly recalcitrant to treatment with current antibiotics.
 
 
CRISPR/Cas-9 Mediated Knock-out of CD147 in Human Corneal Epithelial Cells   
Marissa Feeley and Pablo Argüeso   
Suffolk University
Poster Session B: 4:00-5:15
 
Collective cell migration is the most widely accepted model of cell migration during wound healing and especially in tumor invasion or metastasis. The displacement of tissue is known to be regulated by dynamic cell-cell and cell-extracellular matrix contacts that allow the cells to move together as a whole. Cluster of differentiation 147 (CD147) or extracellular matrix metalloproteinase inducer (EMMPRIN) is a transmembrane glycoprotein that is widely expressed in human tissue because it plays a central role in cell proliferation, apoptosis, and tumor cell migration, metastasis and differentiation. Previous studies have shown that CD147 promotes cell migration by clustering galectin-3, a lectin implicated in cellular adhesion and differentiation, and promoting the expression of MMPs (matrix metalloproteinases), enzymes capable of degrading many different extracellular matrix proteins. Despite the recent advancements in understanding the functions and interactions of galectin-3 and CD147 in epithelial wound healing, the direct contribution of galectin-3 to the adhesive structures in migrating epithelia remains understudied. Our goal is to identify downstream pathways mediated by galectin-3 clustering of CD147 using CRISPR/Cas-9 technology to knock-out CD147 in human corneal epithelial cells. Understanding the molecular basis of the downstream pathways mediated by galectin-3 and CD147 can reveal how cells migrate and tumors metastasize, thus making this pathway a potential target for cancer therapies and advancements in wound-healing technologies.
 
 
Skeletal fusion and age determination in nonhuman mammals
Vanessa Ferreira-Lopes and Eric W. Dewar
Suffolk University
Poster Session A: 1:00-2:15
 
Skeletal analysis is important for determining the age and identification of human remains when other physical characteristics are beyond recognition. The analysis of the degree of fusion of the skeleton is also important to study life history, age distribution in a population, and to answer other biological questions. The purpose of this study was to determine if other mammals have same age-related patterns of epiphyseal fusion as humans. We compared what is known for humans from the literature with specimens from the Museum of Comparative Zoology to reconstruct trajectories of nonhuman mammals. Our sample included the study of several limb bones: the femur, humerus, tibia, radius, ulna, and pelvis. We found that of among the bones examined, 22% of the skeleton of the white-tailed deer was completely fused during the juvenile life stage. In contrast, we found that in the gray wolf, only 10% of the bones were completely fused in the juvenile stage but the remaining were completely fused by the adult stage. Coyotes showed no fused bones during the juvenile stage but 75% were completely fused at the start of the adult life stage. These results were compared to the human fusion pattern. The literature of forensic anthropology reports that in women 20% of the bones compared are completely fused during juvenile and adult life stages. In men, 60% of the bones are completely fused during juvenile stage and the remaining bones studied are completely fused by the adult stage.
 
 
The Structure of the Retina After Preterm Birth
Lauren Fess1, Emily Swanson2, James Akula2, and Anne Fulton2
1Emmanuel College; 2Boston Children's Hospital & Harvard Medical School           
Poster Session B: 4:00-5:15
 
Retinopathy of prematurity (ROP) is a pediatric neovascular disease that affects the eyes of some prematurely born infants, and is one of the leading causes of blindness and visual impairment in children worldwide. ROP is characterized by abnormal, tortuous blood vessels that can grow out of control and detach the retina. It was hypothesized that premature birth interrupts the normal process of foveal development, permanently disrupting vision. To evaluate if ROP further alters normal processes of foveal development, we analyzed the arrangement cells that form the fovea using optical coherence tomography (OCT) and adaptive optics scanning light ophthalmoscopy (SLO). We also measured visual acuity (VA). We tested 93 former preterms (43 with No ROP, 34 with Mild ROP that did not require treatment, and 16 with Severe ROP that required laser ablative therapy) and 89 term-born controls. We exported OCT layer boundaries to a custom program. We measured depth, breadth, and layer thickness. Finally, we identified individual cones in the SLO. We found that increased severity of ROP was associated with decreased foveal depth and VA. The shallower pits in preterms were due to thicker postreceptor laminae; ONL thickness was not associated with VA. Residual, severe ROP foveae were broader than normal and, in these subjects, breadth predicted VA. Cone distributions in preterm retinae displayed more irregularity and apparent lower density. Cone spacing and ONL thickness were correlated. These data suggest that ROP further interrupts the normal process of foveal development and that parameterization of foveal structures can provide quantitative descriptors of retinal abnormalities.
 
 
Impact of benzyl butyl phthalate exposure on Hyalella azteca morbidity and mortality
Alexandra Fidalgo and Lisa A. E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
            Benzyl butyl phthalate (BBP) is a phthalate plasticizer used to make plastics more pliable. Phthalate plasticizers readily penetrate soil which, then contaminates ground water and nearby water ways where it can come in contact with aquatic organisms such as H. azteca, an abundant North American crustacean amphipod. The objective of this pilot study was to identify the concentrations of BBP necessary to induce morbidity and death in a laboratory population of H. azteca. Since BBP is used with a chemical vehicle, it was also necessary to determine what effect, if any, the vehicle acetone had on the population. Organisms were exposed to BBP concentrations ranging from 0.25 µg/L to 10.00 𝜇g/L. At 12, 24, 36, and 48-hour intervals, organisms were examined and classified as healthy, morbid, or dead. Over the entire 48-hour observation period, 95% of H. azteca controls remained healthy with only 5% exhibiting morbidity and 0% mortality. In groups exposed to the acetone vehicle, health declined to 62.9%, and morbidity and mortality increased to 32.9% and 5.4%, respectively. With low BBP doses (0.25 µg/L to 2 µg/L), the acetone effect appeared to be reduced indicating a possible antagonistic chemical relationship. Moderate BBP doses (3 µg/L to 6 µg/L) resulted in H. azteca health and morbidity that mirrored that of acetone alone. High BBP concentrations (6 µg/L -10 µg/L), however, eclipsed all other treatment groups in terms of declining health (49%) and increased morbidity (48%). Overall, these results indicate that the acetone vehicle reduced health and increased both morbidity and mortality in H. azteca. They also suggest the possibility of a dose-dependent interaction between BBP and the acetone vehicle which effects morbidity and mortality endpoints. Future studies will focus on clarifying this possible chemical relationship.
 
 
Impact of Benzyl Butyl Phthalate on the Neuroendocrine System of Procambarus clarkii
Maria Figetakis and Lisa A. E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
Benzyl butyl phthalate (BBP), a plasticizer involved in polyvinyl chloride production, is able to seep out of plastic material and contaminate water, soil, and air. It is a potential endocrine disrupter and may have an effect on the central nervous system, resulting in an alteration of animal behavior. The purpose of this experiment was to observe the effect of BBP exposure on crayfish (Procambarus clarkii) ecdysis as well as brain dopamine and serotonin. Crayfish were randomly assigned to one of the following treatment groups: unexposed (C), acetone exposed (A), ethanol exposed (E), BBP in acetone (BA), or BBP in ethanol (BE). All dosing, regardless of chemical, was 0.1ppm every seventy-two hours for a total of eight weeks. Ecdysis appeared to be dependent on treatment with the greatest incidence occurring in control and the least among those exposed to ethanol (80% C >100% BA* >80% A >60% E and 60% BE). Mortality among all groups appeared to be dependent on treatment with the highest mortality occurring in BA and the lowest among C (60% BA < 33% A <20% BE < 17% E <0% C). Serotonin and dopamine ELISAs were performed on fresh brain tissue homogenate, and although both neurotransmitters were detected, there was no significant difference in either dopamine or serotonin among treatment groups. These results indicate that there may be endocrine disruption impacting crayfish ecdysis, but the disruption does not appear to involve either serotonin or dopamine.
 
 
Cell-cell adhesion pattern along the cell cycle         
Bindika Ghimire and Phuong Nguyen
Colby-Sawyer College            
Poster Session A: 1:00-2:15
 
Cell-cell adhesion and cell cycle are two critical process in any multicellular organism. Cell-cell adhesion is one of the way the cells communicate and the cell cycle is a process in which the cells proliferate. Because of the importance of these two processes in the existence and the development of multicellular organisms, we wanted to investigate the relationship between cell-cell adhesion and the cell cycle. More particularly, we focused on examining the cell-cell adhesion pattern along the cell cycle. We used the aggregation assay to separate the population of mouse epithelial cell that is more adhesive from the one that is less adhesive. We quantified the cell cycle distribution in two populations using the fluorescence intrinsically labeled indicating phase of the cell cycle. The study was verified by aggregation performed in cell cycle synchronized population. We found that at G1 phase of the cell cycle the cells were more adhesive while at G2 phase the cells were less adhesive. We think that G1 phase might be very important for determining the characteristics as well as the fate of the cells, therefore, the cell communication is stronger at this phase.
 
 
Isolation and characterization of antibiotic producing Streptomyces griseus strain JG-2 from inner city soil      
Jayda Gilmore and Jessica A. Smith
American International College         
Poster Session A: 1:00-2:15
 
Scientists are constantly looking to discover new antibiotics due to the rise of antibiotic resistance in hospitals, communities, and the environment. Soil microorganisms steadily evolve to produce novel antibiotics in order to kill off their competitors. To discover an antibiotic-producing microorganism, a sample of soil was collected and bacteria were enriched for. One strain, JG-2, was isolated and was found to release antimicrobials that were effective against a wide range of both gram-positive and gram-negative bacteria. JG-2 is a gram-positive bacillus, which is able to withstand many harsh environments including cold temperatures and high salt concentrations. Further studies on JG-2 will provide insight as to how this organism can withstand extreme environments and also produce antibiotics.
 
 
Differences in Pediatric Gastrointestinal Patients: Autism Spectrum Disorders versus Control
Saige Greenwell1, S. Ballal2, and R. Lite1
1Simmons College; 2Boston Children's Hospital        
3:00–SMT 315
 
Autism Spectrum Disorders (ASD) is a term which encompasses a wide variety of disorders such as Rett’s Disorder, Asperger’s syndrome, and pervasive developmental disorder not otherwise specified (PDD-NOS), among others. Children with ASD exhibit a higher rate of gastrointestinal (GI) symptoms than children without ASD. This study aims to investigate whether or not patients at Boston Children’s Hospital (BCH) who have ASD exhibit anesthesia and/or outcome related differences for GI procedures as compared to control patients without an ASD diagnosis. More specifically, it will explore whether or not patients could receive IV sedation without inhaled anesthesia induction, and how long they were under anesthesia for. Other variables studied will include differences in histological findings during procedures between the two groups and differences in procedural indications. This study is a retrospective chart review; data was collected from patient charts at BCH using Children’s 360 and PowerChart software. Only subjects who received an esophagogastroduodenoscopy (EGD) in 2016 or early 2017 or a colonoscopy during 2015 or early 2016 were included. It is important to study these differences in order to help physicians to provide better treatment for children with ASD moving forward.
 
 
Benzyl butyl phthalate modulation of brain serotonin and dopamine in Fundulus heteroclitus (mummichog)
Breanna Guglielmi, Andrea Leboeuf, Matthew Moeller, Allie Deegan, and Lisa A. E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
This study was designed to determine the effect of benzyl butyl phthalate (BBP) and its ethanol vehicle on Fundulus heteroclitus behavior and swim pattern. Fish (N=38) were randomly assigned to one of three groups: untreated, ethanol, and BBP. Fish were exposed daily to 0.1 ppm ethanol vehicle or 0.1 ppm BBP-vehicle combination for 23 consecutive days. Fish were fed twice daily, and at each feeding, the number of lethargic and active (normal) fish were recorded. On days 0 (T0), 14 (T14), and 24 (T24), each fish was placed in an observation chamber superficially divided into nine sectors, and sector changes within a two-minute interval were recorded.
At feeding time, the majority of fish within both the ethanol-exposed and control groups exhibited normal behavior (P≤ 0.0001). Among BBP-exposed fish, however, normal and lethargic behavior was equally divided. More ethanol-exposed fish displayed normal behavior than control (66% vs. 60%, P=0.0003), and BBP-exposed fish were significantly more lethargic at feeding time (53%, P= P=0.0016) than the ethanol (34%) or unexposed (40%) groups (P=0.0002).
At T0, 45% of unexposed fish demonstrated normal swim activity, and 50% appeared agitated. By T14, the number of unexposed fish exhibiting normal swim activity had increased to 50%; no subsequent change in the ratio of normal/lethargic swim activity was observed between T14 and T24. Swim pattern was, therefore, useful in indicating acclimation to captive housing.
At T14, ethanol-exposed fish displayed normal swim patterns less frequently (29%) than lethargic (71%). At T24, the incidence of normal swim pattern rose to 43%, while lethargic fell to 43% and 14% grew agitated. Interestingly, swim pattern alteration in response ethanol exposure appears to be dose-dependent with initial exposure inducing greater activity (T14) and longer exposure (T24) resulting in lethargy and agitation.
At T14, 14% of BBP-exposed fish exhibited normal swim patterns, and 57% were lethargic. By T24, 57% were normal and 29% were lethargic. These observations did not, however, differ significantly from ethanol-exposed control. Since BBP changed behavior at feeding time but did not alter swim pattern (as compared to control), behavior at feeding time is a more appropriate endpoint than swim pattern.
 
 
Dysfunction of Retromer Complex and its Association with Neurodegenerative Diseases           
Sara Haque1, C. Petit2, T. Walther2, C. Russell1, and J. Lopilato1
1Simmons College; 2Harvard T. H. Chan School of Public Health       
Poster Session A: 1:00-2:15
 
Debilitating neurodegenerative conditions such as Alzheimer’s, Parkinson’s, and Lou Gehrig’s Diseases take away the functionality of loved ones, cherished childhood memories, and leave a lasting impact on social relationships. A common feature of these diseases is the loss of functioning neurons from all neuronal classes. It therefore becomes essential to conduct further research to establish, at the molecular and cellular levels, the origins of neuronal death which will serve as potential treatment targets. One possible mechanism by which neurodegeneration evolves is through the defective retromer complex. Located on endosomal membranes, the retromer complex regulates the retrograde transport of proteins, including neurotransmitters and lysosomal proteases, to their recycled destinations. Several mutations in the retromer complex have been identified both in Alzheimer’s and Parkinson’s Diseases, specifically, in the vacuolar protein sorting-associated protein 35 (VPS35) subunit of the retromer protein, making it the primary subunit of interest in this study. VPS35 plays a central role in the binding and transporting of material between the endosome and various destinations. It is hypothesized that cells with mutated or absent retromer complexes will display elevated levels of lysosomes, in size and quantity, due to the accumulation of substances requiring degradation. Data confirmed that a heightened level of lysosomes is seen in retromer defective cells, suggesting altered retrograde trafficking. It is also shown that sphingolipid concentration is elevated, possibly associated with the lack of elimination of molecules within lysosomes. Future implications for this study include monitoring a long-term cohort of retromer-mutated mice to record prolonged phenotypical effects.
 
 
DMD-10 and its Potential Effect on Regulation of the Glutamate Receptor GLR-1 in C. elegans 
Lily Johnsky and Annette McGehee   
Suffolk University       
Poster Session B: 4:00-5:15
 
An organism’s nervous system processes decisions on how to respond to their environment. Studying how proteins of the nervous system are regulated enables us to identify how an organism responds to its environment and alters its behavior. In our lab, we study glutamate receptors, specifically GLR-1. Glutamate is used for communication between neurons, and changes in glutamate receptors are believed to play an important role in learning and memory. The DAF-7/TGF-ß signaling pathway regulates GLR-1 in C. elegans. However, the mechanism through which GLR-1 levels are increased in the daf-7 mutant worms is unknown. My research addressed how GLR-1 levels were increased in the daf-7 worms. I explored the possibility that the transcription factor DMD-10 is affected in the daf-7 mutants, and then in turn affects GLR-1. We tested this by measuring spontaneous reversals (a GLR-1 dependent behavior) in both single mutant and double mutant worms. If DMD-10 is affected in the daf-7 mutants, we predict that the dmd-10; daf-7 double mutants would have fewer reversals than the daf-7 mutant. The dmd-10; daf-7 double mutants did not have fewer spontaneous reversals than the single daf-7 mutant, which suggests that the DAF-7/TGF-ß signaling pathway does not use DMD-10 to alter levels of GLR-1. However, the dmd-10 single mutant had fewer reversals than the wildtype, suggesting that DMD-10 is required for normal reversals. The results of this experiment suggested that although DMD-10 is not involved in DAF-7/TGF-ß regulation of GLR-1, it may have an independent effect on GLR-1. My current research is aimed at determining if DMD-10 is involved in the regulation of GLR-1.  I have generated a strain of dmd-10 mutant worms that expresses GFP-tagged GLR-1 (GLR-1::GFP). Imaging of GLR-1::GFP in wild type and dmd-10 mutant worms will allow us to test if DMD-10 is involved in GLR-1 regulation.
 
                                                           
Isolation of a Pseudomonad that inhibits growth of Staphylococcus aureus          
Kristin Kazzi, E. Childs, and C. Berkes
Merrimack College
Poster Session B: 4:00-5:15
 
Nature is a rich reservoir of antibiotics.  Soils in particular are rich in bacteria and fungi that produce antibiotics enabling them to interfere with the growth of competing species.  Here, we aimed to isolate and characterize antibiotic-producing microbes from soils collected on and near the Merrimack College campus. An array of environmental soil samples were collected and tested for any antibiotic properties against Pseudomonas aeruginosa, Candida albicans, Staphylococcus aureus and Escherichia coli. We have identified a soil isolate – “SDA1yellow” – that displays significant inhibition of S. aureus growth.  To identify “SDA1yellow”, we subjected it to Gram staining, biochemical analysis, and 16S rRNA sequencing.  Taken together, our results identify SDA1yellow as a member of the Pseudomonas genus.  To identify the antibiotic compound, we are currently performing organic extractions of culture supernatants for analysis by LC-MS. Additional testing is also underway to identify the species identity of our isolate. 
 
 
Evaluating the effectiveness of utilizing morphological characteristics in a neural network to classify fish gut flora
Alexander Kirst, Jonathan Blake, Mark Hoffman, and Lisa A.E. Kaplan
Quinnipiac University
Poster Session A: 1:00-2:15
 
Identification of an organism’s residential gut flora can be a time consuming and costly proposition involving purification, PCR, sequencing, and BLAST searches. The focus of this project is to reduce both cost and time by developing a neural network to identify microbes based on their morphology. A Python-based neural network library was developed and used to classify F. heteroclitus gut flora based on morphological characteristics ascribed to the microbial colony (color, size, appearance, and convexity) as well as those associated with the individual bacterial cells (shape and gram stain). Using previously identified samples with known morphological data, this project aims to train a neural network for use in identifying unknown bacteria samples. The results of this study indicate that the network is accurate at predicting the identity of bacteria within the Vibrionaceae and Flavobacteriaceae families. While additional data is needed to expand predictions to other families, development and application of this neural network should save time and limit the expense associated with PCR, generating 16S ribosomal sequences, and BLAST searches.
 
 
A Comparison of Culture Techniques for the Isolation and Identification of Gut Flora Diversity in Fundulus heteroclitus           
Hannah Kissinger, M. Figetakis, A.L. Luchini, M. Steinle, J. Mital, J. Blake, and L.A.E. Kaplan
Quinnipiac University
Poster Session A: 1:00-2:15
 
Fundulus heteroclitus (N=20) were collected from a sand spit-barrier beach at the Milford Point Long Island Sound Estuary in September of 2016.   The fish were transported to the laboratory and immediately sacrificed.  Intestinal contents were plated on Marine agar (N=10) or Tryptic Soy Agar (TSA) (N=10) culture plates.  Bacterial colonies were isolated and morphologically identified.  All confirmed isolates were selected for polymerase chain reaction (PCR) and genetic sequencing of the 16S ribosomal unit. Colonies isolated on marine agar had significantly (p=0.05) higher survival rate than TSA isolates (H. Kissinger, et. al. 2016). Of 96 marine agar isolates, 71 identities were confirmed through a double-blind analysis using BLAST search queries and the algorithm of Gotoh (1982).  Of the remaining 25 isolates, 24 isolate identities were in agreement at the genera level but not at the species level. One isolate had no homology found by either method.  Overall, morphological comparison failed to determine the identities of 2 isolates and confirmed the identities of 69 isolates.  Isolate identification for marine agar and TSA experimental groups is ongoing.
 
 
The Effectiveness of Auranofin and Related Gold (I) Compounds as Antimicrobial Agents           
Bianca Kun1, Janet B. Foley2, and Cassandra Saitow1          
1Simmons College, 2Bennington College
Poster Session B: 4:00-5:15
 
Today’s antimicrobial drugs are less effective due to the evolution of resistant bacteria. This problem has increased the need to develop novel antimicrobial agents that would combat these microorganisms through an alternative mechanistic pathway. Recent publications have reported results from a high-throughput screening identifying the anti-rheumatic drug auranofin as an effective antimicrobial compound. The structure of auranofin contains a gold (I) thiolate which is hypothesized to cause death in bacteria by inhibiting the thioredoxin pathway. A small library of gold (I) thiol compounds and gold nanoparticles (AuNP), developed by the Foley Lab, with chemical structures related to auranofin, will be analyzed for antimicrobial activity in order to identify specific chemical moieties responsible. These compounds will be tested against the bacteria Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterobacter aerogenes, Staphylococcus aureus, and Enterococcus faecium. These six bacteria were chosen because they are variants of common nosocomial pathogens. They were assayed against auranofin and related compounds in a toxicity assay to determine the efficacy of the compounds by measuring the inhibition of bacterial growth. Preliminary studies show differential response corresponding to chemical variations. Data from this study will potentially show a structure-function relationship among gold compounds which are most effective against bacteria. This information could guide development of new gold-based antimicrobials.
 
 
Impact of Urbanization on the Muddy River, Boston, MA
Kate LaGattuta1, Danny Ly1, Ian Chamenko2, Tala Ferguson1, and Lisa Lobel1
1Wheelock College; 2Emmanuel College       
Poster Session B: 4:00-5:15
 
Continuous temperature and conductivity data were collected within the Muddy River to assess daily and seasonal changes. Understanding the past levels and natural variations in these parameters will aid in determining improvements resulting from the restoration project as well as long-term changes due to climate change. In particular, we aim to characterize the background conductivity levels found within the river and how this is impacted by the use of road salt as a deicer. Temperature and conductivity were monitored hourly for two years in the Muddy River. Lettuce seed bioassays conducted on Muddy River water samples were used to determine if high conductivity had a negative impact on seed germination. Summer water temperatures exceeded the Massachusetts water quality standard in both years. Conductivity spikes as high as 9,000 µS/cm were associated with winter precipitation events while decreasing conductivity levels were associated with summer precipitation events. These data will provide valuable background information to better understand the intersection between the restoration efforts and climate change impacts within this small urban watershed.
 
 
The role of intergenic regions in regulating RpoS    
Adrian Lambert, Cheikh Diop, and Celeste Peterson
Suffolk University
Poster Session A: 1:00-2:15
 
E. coli express the rpoS gene when subject to stress. RpoS is responsible for activating hundreds of genes which make E. coli dormant and capable of surviving harsh conditions. This gene is regulated transcriptionally by numerous transcription factors and post-transcriptionally regulated by several small noncoding RNAs (sRNAs). However, not all the genes that regulated rpoS have been identified. We carried out an overexpression screen to identify novel regulators of rpoS. We discovered a region between the glnS and the chiP genes that down-regulated rpoS when overexpressed as measured by Western blot and rpoS’- ‘LacZ fusions. Data from additional reporter fusions (a short rpoS’- ‘lacZ translation fusion and an rpoS -lacZ+ operon fusion) indicated that this region decreased rpoS translation but not transcription. To investigate the relationship between rpoS and the glns-chiP intergenic region, an E. coli strain lacking the 5’UTR of chip was cloned. This new strain was used to prove that the 5’UTR of chiP is needed for rpoS downregulation. Thus the 5’ UTR of a gene can regulate rpoS levels, either directly or indirectly. The chiP gene was then induced by addition of chitobiose to the cell cultures in both LB and Minimal media to see how rpoS levels would be affected. It was found that induction of chiP by chitobiose did not affect the rpoS levels in either media. In summary, intergenic regions may affect rpoS expression when present in high copy numbers.
 
 
Effect of Plant Extracts on Lactic Acid Fermenting Bacteria found in the Oral Cavity
Monica Liang and Dawn Holmes
Western New England University
Poster Session B: 4:00-5:15
 
            Lactic acid fermenting bacteria are the primary organisms involved in plaque formation and tooth decay. Therefore, compounds that can inhibit the growth of these bacteria have important applications within the field of dentistry. In this study, two lactic acid fermenters found in the oral cavity, Lactococcus lactis and Streptococcus mutans, were cultured on tryptic soy agar and exposed to various plant extracts in order to observe antimicrobial activity. Twelve different extracts were tested, including α-pinene, β-carophyllene, anisaldehyde, linalool, methyl anthranilate, 6-methyl-5-heptan-2-one, 1,2,4-trimethoxybenzene, peppermint, spearmint, clove, lemon, and cinnamon. Results from disc diffusion assays showed that S. mutans was most sensitive to anisaldehyde and 6-methyl-5-heptan-2-one with zones of inhibition as large as 2.0 cm and 1.8 cm, respectively. L. lactis was most sensitive to linalool and 6-methyl-5-heptan-2-one, which both yielded zones of inhibition of 2.0 cm. While α-pinene slightly inhibited growth of L. lactis, it had no effect on the growth of S. mutans. Spearmint extract only slightly inhibited both L. lactis and S. mutans and exposure to this compound resulted in zones of inhibition that were < 1.0 cm in diameter. These results are significant and should be taken into consideration when developing new toothpastes or mouthwashes designed to help fight tooth decay.
 
 
The effect of Dominant hemimelia on mouse mid- and hindgut development     
Kelsey Lipiner and M. Owen
Simmons College
Poster Session B: 4:00-5:15
 
Formation of the primitive streak, in the gastrulation stage of early embryogenesis, establishes the major body axes of the mammalian embryo, determining placement, orientation and formation of organs. The mouse mutation, Dominant hemimelia (Dh), produces defects in this patterning serving as a model for study of processes that govern development of organs along the anterior-posterior (A-P) and left-right (L-R). Animals carrying the Dh mutation exhibit reduced numbers of lumbar vertebrae, asymmetric preaxial hindlimb defects and asplenia. Additionally, Dh mice have been reported to have smaller stomachs and shortened guts. These phenotypic effects suggest a primary defect in A-P and L-R patterning of tissues formed from, or induced by, the mesoderm. Abnormalities in the gut and hindlimbs may result from an effect of Dh on the temporal-spatial expression of Hox genes in the mesoderm. This study was undertaken to further characterize mid- and hindgut defects, their association with skeletal malformations, and the expression of Hoxd 10-12 in the developing gut and hindlimbs of Dh embryos and their wild- type (+/+) littermates. Preliminary analyses reveal a shortening of the gut of Dh, as compared to +/+ littermates, with smaller or absent ceca, in correlation with more severe skeletal defects. In situ hybridization studies are in progress; differences in expression domains of Hoxd 10-12 in Dh and +/+ animals would provide insight into the role of this gene in patterning of the mammalian vertebrate embryo.
 
 
Survey of Methylobacteria Species Associated with Mycorrhizal, Saprophytic, and Parasitic Fungi Found in Southeastern Massachusetts     
Kristen Lowder, Ashlyn Czapiga, Alexander Crowley, Magdalena James-Pederson, and Rachel Hirst
Stonehill College
Poster Session A: 1:00-2:15
 
Methylobacteria are pink-pigmented facultative methylotrophs capable of utilizing methanol as their sole carbon source.  Methylobacteria are commonly found associated with plants in symbiotic relationships, in which the plant releases methanol during gas exchange through stomata and the bacteria provide phytohormones that aid in plant growth and development.  While studying lignin degradation enzymes in the white-rot fungus Armillaria gallica, an isolate from the Methylobacterium genus, now deemed Methylobacterium sp. ARG-1, was characterized.  This was the first published association between Methylobacteria and a white-rot fungus. 
Expanding upon past research, this project focused on broadening the analysis to determine whether Methylobacteria naturally associate with other types of fungi.  41 fungal samples, including mycorrhizal, saprophytic, and parasitic mushroom-forming fungi from 19 different genera, were sampled.  Using 16S rRNA gene sequencing, it was determined that each fungal sample contained at least one associated species of Methylobacteria.  In addition to suggesting a widespread association between Methylobacteria and fungi, our results indicate that communities of Methylobacteria associate with fungi, and these associations may be based on fungal ecology.  Further research is necessary to elucidate potential biochemical explanations for the association between Methylobacteria and fungi from mycorrhizal, saprophytic, and parasitic ecologies.
 
 
Quantifying episodic memories from real-world experience          
Alyssa Marconi1 and Gabriel Kreiman2
1Emmanuel College; 2Children's Hospital, Harvard Medical School 
Poster Session B: 4:00-5:15
 
Episodic memories form the fundamental fabric of who we are. These memories are full of content and malleable due to the number of inputs impinging upon our senses in our everyday lives. Despite a plethora of laboratory studies about memories for individual words, names, or pictures, little is known about what governs the formation of episodic memories in real life. In this work, we studied real life episodic memories by recording ground truth video and eye movement information during a one hour segment in the life of our subjects and subsequently evaluating their ability to recognize those events. Ten subjects completed an encoding task which took place in two versions: an outdoor and indoor route. Memory recognition tasks were completed at 24 hours up to 3 months post-encoding and performance was consistent across subjects in both versions with an accuracy of up to 62 +/- 7.0%. Previous work used a similar methodology combined with machine learning approaches to predict episodic memory formation in movies, but those results remain inconclusive since movies are artificial stimuli and do not accurately represent real life events. Our study found that real life episodic memories are less memorable than movies likely due to the large amounts of information we filter out in non-laboratory settings. The results allow us to understand episodic memory formation in real life to elucidate the subjective filtering which leads to how we learn and consolidate long-term memories. To evaluate the content properties that influence long-term memory formation, annotations were designed to see what aspects enhance memorability in the real world.  These allow us to report what content properties correlate with higher performance in memory recognition. To the best of our knowledge, this work constitutes the first quantitative approach to directly measure memory formation with ground truth data in real life scenarios.
 
 
Isolation and characterization of antibiotic, metal, and chemical sensitive Staphylococcus epidermidis stain MM-10   
Meagan McCarthy and Jessica A. Smith
American International College
Poster Session A: 1:00-2:15
 
Staphylococcus epidermidis is a ubiquitous colonizer of human skin and plays an important role in balancing the microflora of epithelial tissue. S. epidermidis rarely becomes pathogenic, however many strains carry genes for antibiotic as well as chemical resistance. Strain MM-10 of S. epidermidis was isolated, and found to be a gram-positive staphylococcus, which was halotolerant, mesophilic, and a facultative anaerobe. 16S rRNA sequencing showed that strain MM-10 is closely related to many other strains of S. epidermidis. Strain MM-10 is unique in that it was sensitive to all antibiotics, chemicals, and heavy metals tested, including penicillin, even after extended incubation. Identifying strains of Staphylococcus species that are sensitive to antibiotics such as penicillin is rare since Staphylococcus species are known for developing resistance to antibiotics. Further studies on this species could be useful in understanding the modes of antibiotic resistance.
 
 
An investigation into the role of IDA-1 in DAF-7-dependent regulation of GLR-1  
Shannon McLaughlin and Annette McGehee
Suffolk University
Poster Session B: 4:00-5:15
 
In organisms, behavior can be a reflection of environment and nervous system activity. Previous studies have shown that the DAF-7 signaling pathway in C. elegans is affected by the environment, such as inadequate food supply or high temperature. The DAF-7 signaling pathway also regulates the glutamate receptor GLR-1, and increased amounts of GLR-1 occur in C. elegans strains with mutations in the genes of the DAF-7 signaling pathway. Additionally, spontaneous reversal behavior depends on GLR-1 and increased reversals are evident in the daf-7 mutants. The mechanism by which the DAF-7 pathway affects GLR-1 levels is currently unknown. Our research focuses on understanding this mechanism by investigating the role of the candidate gene ida-1. IDA-1 was identified because of its involvement in regulating presynaptic neurotransmission and its presence in neurons along the ventral nerve cord (VNC) that express GLR-1.To test if IDA-1 was involved in the DAF-7 pathway’s regulation of GLR-1 an assessment of spontaneous reversals in single and double mutants was performed. Results showed there was no significant difference between wildtype and the ida-1; daf-7 double mutants. This result is consistent with a role for IDA-1 controlling GLR-1 in a daf-7 background. To test this fluorescent imaging on worms expressing GFP tagged GLR-1(GLR-1::GFP) will be performed to directly assess GLR-1 levels for each genotype. Preliminary imaging results confirm that GLR-1::GFP is higher in daf-7 than in wild type; in ida-1; daf-7 double mutants GLR-1::GFP levels are similar to wild type rather than to daf-7 single mutants. This result supports the idea that IDA-1 plays a role in the DAF-7 pathway regulation of GLR-1. In the future, quantitative imaging and GLR-1 protein level analysis will be performed to confirm this result.
 
 
The substrates of ClpXP during varying growth conditions in Escherichia coli
Amberly Mendes, Pauline Ngo, Tatjana Von Rosen, and Celeste Peterson 
Suffolk University       
Poster Session A: 1:00-2:15
 
Understanding how bacteria survive during starvation can lead to new strategies for developing antibiotics, especially those that treat dormant bacteria. Bacteria go dormant due to changes in their protein concentration; this can occur at the level of protein synthesis or protein degradation. The goal of this project is to characterize candidate substrates for the protease ClpXP that were identified in previous experiments and follow their degradation during growth and starvation conditions. Overall, we cloned with an epitope tag and sequence verified a total of seven proteins these include GatY, GltX, AdhP, MalT, TnaA, KatE and Ndh. We successfully visualized two proteins on a Western Blot, GltX and GyrA (an untagged version). Results from this work will lead to a better understanding of how the cell regulates its degradation processes during starvation and dormancy.
 
 
Daily Occurrence of Operophtera (Lepidoptera: Geometridae) in Carlisle, MA from 2012-2016
Caroline Mitchell and Peter R. Burn  
Suffolk University       
Poster Session B: 4:00-5:15
 
As part of a larger study of moth seasonality, members of the genus Operophtera were counted nightly from photographs taken between 2012 and 2016 at the porch lights of a residence in Carlisle, MA.  The two local species of Operophtera are O. brumata (the Winter Moth) and O. bruceata (the Bruce Spanworm), but the two are indistinguishable from photographs.  The pattern of seasonal occurrence was similar in all years, with animals appearing in early to mid-November and disappearing by mid to late-December.  The total numbers by year were 1046 (2012), 1032 (2013), 1385 (2014), 3306 (2015), and 1290 (2016) for a total of 8059 animals observed.  The daily data suggest two unequal peaks of occurrence, possibly corresponding to the two species.  The daily differences in occurrence were extreme, with more than 1000 moths counted on one night, and more than 46% of the total during the top 10.  The nights of higher moth abundance corresponded to warmer temperatures, but the data suggest more a threshold than a linear relationship.  The presence of extreme daily differences in occurrence suggests that ecological patterns for such species need be observed at shorter or more frequent time intervals than has frequently been the case.  
 
 
A novel transcription factor regulates the oxidative stress response in in Caulobacter crescentus          
Nasser Mohieddin, Nick Maragos, Brandon Smolarek, Arup Dey, and Celeste Peterson    
Suffolk University
3:00–SMT 316
 
All aerobically growing organisms have developed mechanisms for coping with oxidative stress. While the oxidative stress response has been well-studied in some bacteria such as Escherichia coli, it is not as well-characterized in other bacterial groups such as alphaproteobacteria. Here we examine the role of transcription factors in the oxidative stress response in Caulobacter crescentus. Using viability assays, we find that a deletion in the transcription factor CoxR leads to a strain that is more sensitive to hydrogen peroxide treatment but not to heat shock and other DNA damaging agents such mitomycin C. To find the genes that CoxR regulates, RNA-seq analysis was carried out in the wild type and CoxR deletion strains. Results suggested that the CoxR deletion strain has altered gene expression of NADPH cycling enzymes such as ferredoxin, ferredoxin reductase and cytochrome P450. To confirm the RNA-seq data, we carried out qRT-PCR and ferredoxin reductase assays. Our results showed that mRNA levels of these NADPH cycling genes and ferredoxin reducatase activity were higher in the CoxR deletion strain. In summary, CoxR is critical for the oxidative stress response and controls the levels of ferredoxin/ferredoxin reducatase/Cytochrome P450.
 
 
Impact of benzyl butyl phthalate and its ethanol vehicle on F. heteroclitus feeding behavior and locomotive patterns
Matthew Moeller, Allie Deegan, Harry Pylypiw, and Lisa A.E. Kaplan         
Quinnipiac University
Poster Session A: 1:00-2:15
 
This study was designed to determine the impact of benzyl butyl phthalate (BBP), a plasticizer and suspected EDC, on F. heteroclitus serotonin (5-hydroxytryptamine, 5H-T) and dopamine neurotransmitters. Fish were assigned to one of three groups: unexposed control, ethanol vehicle control, or BBP (delivered in an ethanol vehicle). Fish were dosed daily with either ethanol or BBP in ethanol (0.1 mg/L) for 28 consecutive days. Serotonin and dopamine levels were quantified at the end of two and four weeks of dosing as well as at the end of a 2-week recovery period during which fish received neither ethanol nor BBP.
The two-week ethanol exposure resulted in no significant difference in dopamine, but did produce significantly higher serotonin (113%, P=0.0048) as compared to unexposed controls. Following a four-week ethanol exposure, no significant difference was observed for dopamine, but there was a 41% reduction in serotonin (P=0.0018) as compared to control. While no significant difference in serotonin was observed between ethanol exposed and control fish at the end of a two-week recovery period, a significant (P=0.0001) 5% reduction in dopamine was observed.
The two-week BBP exposure significantly reduced both dopamine (35%, P=0.046) and serotonin (91%, P=0.05) as compared to the ethanol vehicle control. No significance differences, however, were observed in dopamine or serotonin after a four-week BBP exposure or following the 2-week recovery period. Overall, these results indicate that ethanol and BBP modulate serotonin and dopamine in F. heteroclitus as early as two-weeks exposure. It also appears that a two-week recovery period was insufficient to completely eliminate differences in serotonin and dopamine among ethanol, BBP in ethanol, and non-exposed individuals.  Future experiments will focus on the recovery aspects of exposure.
 
 
An analysis of phosphorus levels in the Charles River from 1996 to 2015 in relation to policy enactment          
Kayla Myros               
Simmons College        
Poster Session A: 1:00-2:15
 
The phosphorus level in rivers is significantly affected by stormwater polluted with impervious surface runoff and decomposing organic matter from urbanized areas. Elevated phosphorus levels in waterways contributes to eutrophication and hypoxia from algal blooms. The purpose of this case study is to assess the importance of stormwater pollution reduction policy on waterways, specifically analyzing the Charles River’s phosphorus levels between 1996 to 2015. I hypothesize that with the introduction of federal wet weather regulations in Massachusetts, the phosphorus concentrations in the Charles River will have decreased overall from 1996 to 2015. I also expect the levels to fluctuate with seasonal changes: rising in the summer and falling in the winter because of increased fertilizer use and detritus waste in the summer. I analyzed quarterly water quality data, collected by the Charles River Watershed Association, using a repeated measures statistical test. The repeated measures test quantifies the variable’s change over time and the statistical significance of that change. Based on the overall trends, the study ends with a commentary on implementing infrastructure and local practice changes that will reduce pollution infiltration and improve water quality.
 
 
Investigation of Transgenerational Immune Priming on Bacterial Clearance and Immune Protein Regulation    
Brittany Noonan1, N. Hays1, B. Gezahegn2, K. Thwin1, A. Ruditsky1, M. Ross1, J. Ramseyer1, W. Smith1, B. Rosengaus1 
1Northeastern University; 2Baldwin Wallace College
Poster Session A: 1:00-2:15
 
It has previously been thought that insect immune systems were non-adaptable, consisting solely of innate immune responses. However, it has recently been discovered that, despite lacking antibodies, insect immune responses can be “primed”; therefore exposure to a pathogen results in improved defense against that pathogen in the future. Additionally, it has been observed that survival benefits may extend across generations. Offspring whose mothers are exposed to pathogens exhibit increased resistance to these pathogens, a phenomenon known as transgenerational immunity. In the present study, we examined possible transgenerational immune responses in the tobacco hornworm, Manduca sexta. Females were injected at the pupal stage with live or heat-killed bacteria, Serratia marcescens. Controls received either no treatment, or were injected with sterile saline. Upon emergence as adults, females were bred with untreated males, and embryos allowed to develop to the first larval stage. We then challenged those larvae with sterile saline (controls) or live bacteria. We predicted that larvae whose mothers were injected with live or heat-killed bacteria would clear live bacteria better than if their mothers were untreated, or treated with sterile saline. We further hypothesized that larvae whose mothers were injected with live bacteria or heat-killed bacteria would up-regulate immune-related proteins such as hemolin. Our results confirmed the hypothesis that maternal exposure to bacteria enhanced the rate of bacterial clearance by offspring. The results were particularly striking for offspring whose mothers had been exposed to live bacteria. Surprisingly, we found that hemolin levels were increased in larvae from all treated mothers, including larvae from saline-injected mothers, relative to those from untreated mothers. These results suggest that the stress of cuticular puncturing is enough for mothers to endow their offspring with enhanced immune competency. Future experiments will explore possible changes in gene expression that may be responsible for insect transgenerational immune responses. 
 
 
Can a lacZ Fusion of cirA detect any effect of BglJ, a Transcriptional Activator?   
Ngozi Nwaoha and Jane Lopilato
Simmons College
Poster Session A: 1:00-2:15
 
The transcriptional activator BglJ activates the cryptic bgl operon of E. coli K12, but the bglJ gene itself is kept silent. The activated bgl operon produces enzymes for the catabolism of aromatic β-glucosides such as salicin. Microarray analysis data showed that the overproduction of BglJ increased the expression of cirA, a gene is required for the uptake of 2,3- dihyroxybenzoyl serine (DHBS), a precursor for the synthesis of the siderophore, enterobactin. Siderophores are used to scavenge iron from host cells. Iron is a required nutrient for bacterial proliferation and the establishment of infection. Thus, its regulation plays a crucial role in the interaction between the host cell and a pathogenic bacterium. The objective of this research is to determine whether a lacZ fusion to cirA will detect any effect of the transcriptional activator, BglJ, on the expression of cirA. The regulatory region of the cirA gene was amplified by PCR and cloned in two bacterial plasmids: pKES268 and pRS1553. The resulting fusion on pKES268 was recombined into the chromosome and assayed for β-galactosidase. The cirA-lacZ fusion on pRS1553 will first be recombined onto phage lambda and then integrated into the chromosome and assayed. These assays will reveal whether there is any effect of BglJ on the cirA-lacZ fusions and whether these two constructs will yield different results.
 
 
Defining Somatosensory Neurons Underlying the Traumatic Peripheral Neuropathic Pain          
Pann H. Nwe1,2, Ling Bai2, Lauren Orefice2, Amanda Zimmerman2, Sreyan Chowdhury2, David Ginty2
1Simmons College, 2Harvard Medical School, Boston, MA    
2:45–SMT 315
 
Approximately 25-50% of all pain clinic visits relate to neuropathic pain. Traumatic peripheral neuropathy is a chronic pain associated with loss of function resulting from physical trauma to the peripheral nervous system. Specifically, allodynia, one of the modalities of traumatic peripheral neuropathic pain, has a profound negative impact on quality life for patients and the difficulty in treatment for this feature of neuropathy remains a major challenge. Recent studies have identified several components causing neuropathic pain such as trophic factors, transduction, central sensitization and immune cell regulation. However, it is still unclear which somatosensory neuron subtypes carrying the pathological pain information to the brain contribute to the development of mechanical allodynia. Here, the role of Aβ Low Threshold Mechanoreceptors (A β -LTMRs) in experimental traumatic peripheral neuropathic pain in the mouse model is explored. Following the spared nerve injury that mimics neuropathic pain, the nocifensive behavioral response is observed by optogenetics activation of Aβ-LTMRs. Our intersectional genetic strategy utilizing the Advillin-Flpo mouse line together with a dual recombinase reporter line, R26-dual-ReaCHR, specifically activated the Aβ-LTMRs that are distinct from classical nociceptors. The data indicate that activation of Aβ-LTMRs induces allodynia-like behavior after the spared nerve injury. Our results can be a starting point to elucidate the relation between Aβ-LTMRs and development of neuropathic pain. The influence of Aβ-LTMRs should be studied further to determine whether or not silencing these populations can block mechanical allodynia.
 
 
Diets of Paleogene mammals tracked more reliably with depositional environment than climate
Cecilia Osimanti and Eric W. Dewar
Suffolk University       
3:30–SMT 317
 
Animals respond to environmental change in modern ecosystems by (1) adapting to new niches through natural selection, (2) migrating to track their habitual niches, or (3) going extinct which can be observed in the fossil record. In this study, we wanted to determine how changes in tooth shape caused by enamel wear can be used to reconstruct diets of species that lived through the Eocene-Oligocene climatic transition (EOCT), a period of cooling and drying from about 38-25 million years ago. During this time, ungulate communities shifted from browsing to grazing as the western grasslands opened up.
We used mesowear analysis of teeth to evaluate the degree of wear of ungulate molars by observing the cusp shape and the relief between these cusps as indicators of diet. This allowed us to classify fossil species into one of three typical diets—fruit and leaf browsing, mixed feeding, and grazing. We studied a sample of casts and photographs of artiodactyl and perissodactyl tooth crowns (n = 594) from of the White River System (WRS) that spans the EOCT in the American West.
Despite the large-scale climatic change that the EOCT is known for, the dietary response by these species when combined into families was minimal. The overall mesowear scores for each of the three time intervals of the EOCT showed no significant difference in overall diets of species in the community (F2,45 = 1.66, P = 0.192), with the exception of Rhinocerotidae (F2,43 = 2.98, P = 0.042).
The sample of the White River species in this study represents the survivors of a number of earlier extinction events. It is possible that the remaining species were better able to endure this climate change interval compared to others that had either gone extinct of migrated elsewhere. The taxonomic makeup of the community was relatively stable across the interval, and the fauna represented the survivors of earlier extinctions, so we think that the generalized niches of theses WRS species were the result of their resilience to extinction.
 
 
The effects of benzyl butyl phthalate on Madagascar Hissing Cockroach neurotransmitters and behavior
Boluwatife Osofisan and L.A.E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
The objective of this study was to determine the effects of benzyl butyl phthalate (BBP), a plasticizer and suspected endocrine disruptor, on Madagascar Hissing Cockroach food consumption, somatic index, and neurotransmitter (serotonin and dopamine) levels. Cockroaches placed into isolation and assigned to unexposed control (N=20) or BBP (N=20) group. BBP was delivered in a 5µL, 30-minute dose via ambient air, three non-consecutive days per week for a total of 6 weeks. Following the 6-week exposure period, cockroaches were decapitated. Homogenate protein concentration was determined and used to standardize the results of the serotonin and dopamine ELISAs results. Although control females consumed two times more food than control males (p=0.04), there was no significant difference in somatic index between control males and females. Dopamine did not differ between control males and females, but control males 80% more serotonin than control females. Food consumption, somatic index, serotonin, and dopamine did not differ between BBP and control males. BBP did, however, increase serotonin (81%) and reduce dopamine (87%) in BBP females as compared to control females. Dosed males had 87% more dopamine than the dosed females, but no significant difference in food consumption, somatic index, and serotonin. These results indicate a baseline difference between male and female Madagascar Hissing Cockroaches. They also suggest that BBP modulation of serotonin and dopamine differs in males and females.
 
 
Unraveling the effect of ER-mediated reactive oxygen species on proteostasis in Caenorhabditis elegans Aging and Neurodegeneration      
Rutvi Patel, Jorge Iván-Castillo Quan, and Keith Blackwell
Joslin Diabetes Center           
Poster Session B: 4:00-5:15
 
Previous work from our laboratory showed that RNA interference (RNAi)-mediated down-regulation of the endoplasmic reticulum (ER) oxidoreductin 1 (ero-1) extended lifespan in C. elegans. Ero-1 allows the formation of disulfide bonds inside the ER lumen by using an inter-cysteine, redox relay that produces hydrogen peroxide. RNAi of ero-1 produces reactive oxygen species (ROS) in the endoplasmic reticulum (ER) that leads to the suppression of the canonical ER stress response pathway. Importantly, when ER stress is induced first, the antioxidant response downstream of SKN-1, is completely inhibited. SKN-1, the major transcriptional regulator of the antioxidant response, is essential for the lifespan extension when ero-1 is down-regulated since the combination of ero-1 RNAi and the absence of SKN-1 led to a much shorter lifespan than controls. ER stress is a driving pathogenic force in age-related diseases, including most of the most common neurodegenerative diseases like Alzheimer’s, Parkinson’s, and Huntington’s disease (HD). PolyQ mutants, which are a representative model of HD- neurodegeneration, have expanded polyglutamine tracts that lead to a loss of motility and protein aggregation. This gave us pause to consider whether ero-1 down-regulation on polyQ mutants would show longevity and healthspan benefits given the presumed underlying ER-stress of an aggregation-prone model. Here we tested the hypothesis that ero-1 RNAi would be detrimental under compromised proteostasis and this would be worsened by the absence of SKN-1. In keeping with our hypothesis ero-1 RNAi increased the number of HD PolyQ aggregates. However, contrary to expected, worms with more aggregates showed improved locomotor ability, an effect we think is linked to SKN-1.
 
 
Investigating the transduction of the infection-associated ospC allele between Borrelia burgdorferi isolates by the bacteriophage fBB-1      
Alexander Preisig and C. Eggers
Quinnipiac University
Poster Session A: 1:00-2:15
 
Bacteria displace genetic material through both vertical transmission from parent cell to daughter cell and through horizontal gene transfer (HGT) between neighboring cells. HGT is thought to play a critical role in the potential virulence of some species known to be pathogenic to humans. The three mechanisms of HGT are conjugation (direct contact between two cells), transformation (uptake of naked DNA), and transduction (via bacteriophage). Borrelia burgdorferi (Bb), the agent behind Lyme disease, is not known to engage in either conjugation or transformation; the species has been shown to transduce one or more bacteriophages collectively known as ϕBB-1. This bacteriophage primarily packages 32-kb circular plasmids (cp32s) and it has been shown previously that the transduction of the cp32s by ϕBB-1 serves as the primary means of HGT and thus genetic variability of this species. A significant plasmid-borne locus has been identified as the ospC gene found on cp26. Alleles of this gene have been linked to causing disease manifestations in human hosts. It has been suggested that this gene could be packaged by ϕBB-1; however, there is little to no evidence to that supports this. The purpose of this work is to investigate whether the ospC gene can be transduced among Bb isolates by ϕBB-1. The results will contribute to the characterization of ϕBB-1 as a mechanism for influencing the infectivity of Bb.
 
 
Small World Initiative at Simmons College: The Implementation of a New Hands-On Research Lab with a Focus on Antibiotic Research and Development 
Madison Rivard and Elizabeth Scott
Simmons College
Poster Session B: 4:00-5:15
 
The antimicrobial resistance crisis is a developing global health threat and new antibiotics are in demand to prevent morbidity and mortalities from typical infections. Soil is the source of over two thirds of antibiotics, and the Small World Initiative (SWI) is an international program that provides a curriculum to engage undergraduates in the search for novel antibiotics in soil. During the academic year of 2016-2017 I facilitated this curriculum at Simmons College. The objective of my work was to identify the best method to adapt the Small World Initiative curriculum for implementation in the Introduction to Microbiology course at Simmons. We sought to identify the most efficient methods of laboratory practices, as well as determine how many antimicrobial producing soil isolates the students might find. I was able to successfully assist in the courses completion by adapting the academic literature, troubleshooting the SWI laboratory procedures, outsourcing certain experiments, and an IRB-approved survey for the students in regard to their opinions on STEM research and development. My work adapting the SWI at this institution allows for involvement in this crowd-sourcing project that provides undergraduate students with a research opportunity and participation in a global effort to tackle the AMR crisis and identify new antibiotics, which could save many lives.
 
 
Effects of Colony composition and isolation on Madagascar Hissing Cockroaches
Sherilyn Rivera, Leah Serunjogi, and Lisa A.E. Kaplan
Quinnipiac University
Poster Session A: 1:00-2:15
 
Habitat disruption changes the dynamics of the physical environment and may lead to isolation of individuals or changes in the composition of the social unit. The objective of this study was to determine the effect of Madagascar Hissing Cockroach social unit disruption on somatic index, food consumption, food preference, and neurotransmitter (serotonin and dopamine) levels. Cockroaches were housed as follows: mixed community (MC) (10 males and 10 females) to serve as control; same-sex community (SS) of 10 males or 10 females; or isolation (ISO) 10 males and 10 females. Following the 6-week exposure, cockroaches were decapitated. Heads were homogenized in 0.1 M PBS; resulting homogenate protein concentration was used to standardize neurotransmitter ELISA results. While no significant differences in food consumption, food preference, somatic index, or serotonin were observed between MC and SS males, MC males had 47% more dopamine (P=0.02) than the SS male group. MC females had 53% more serotonin than the SS female group (P=0.01), but did not significantly differ in food consumption, food preference, somatic index or dopamine. MC and ISO males did not differ in food consumption, food preference, somatic index, serotonin or dopamine. MC females, on the other hand, had a 144% higher somatic index (P=0.0002) than the ISO female group, and 100% (P=0.0001) more serotonin than their ISO female counterparts, but no significant difference in food consumption, food preference, and dopamine. Overall, these results indicate that community composition and isolation impact males by modulating dopamine and females by modulating serotonin.
 
 
Delayed Contribution of Hematopoietically-Derived Central Nervous System Macrophages Utilizing the Myeloablative Compound (Drug) Busulfan
Marina Rocha, Todd Williams, and Josef Kurtz
Emmanuel College
Poster Session A: 1:00-2:15
 
Microglia, the resident macrophages of the central nervous system (CNS), derive from the yolk sac in early development with little contribution from the periphery in the absence of disease.  While CNS inflammation may actively recruit peripheral monocytes in response to disease, it is unclear whether there is significant post-developmental contribution from hematopoietically-derived cells in a non-disease state.  To study this contribution of hematopoietically-derived cells we established mixed syngeneic chimeric mice utilizing the myeloablative drug Busulfan followed by a hematopoietic stem cell transplant (HSCT) of GFP+ donor cells into C57BL/6 recipient mice.  Recipient mice were administered Busulfan (30mg/kg body weight) on days 7, 5 and 3 prior to a transplantation of hematopoietic stem cells from C57BL/6-Tg (UBC-GFP) donor mice.  Blood chimerism levels were analyzed through flow cytometry and yielded greater than 90% donor-derived monocytes by 5 weeks post-HSCT.  Beginning at 8 weeks post-HSCT, ~2-3% of microglia (CD11b+ CD45mid) were donor-derived (GFP+) and ~60% expressed the immune protein MHC II, suggesting recent engraftment and an immune phenotype.  This established CNS chimerism remained relatively stable long-term, as ~1-3% of microglia were donor-derived (GFP+) across later time points, including at 14, 33, and 39 weeks post-HSCT.  Supporting immunofluorescence evidence of donor-derived microglia was observed by Iba-1+ GFP+ MHC II+ non-ramified cells in the choroid plexus as well as Iba-1+ GFP+ MHC II- ramified cells in the brain parenchyma.  To study how increased stress to the hematopoietic system affects donor cell contribution to the resident microglial population, chimeric mice (24 weeks post-HSCT) were treated with additional Busulfan (25 mg/kg body weight on two separate days).  At 15 weeks post-HSCT, an increase to ~3-5% CD11b+ CD45mid GFP+ CNS cells was observed.  Taken together, these data support a contribution of hematopoietically-derived precursor cells to the resident CNS macrophage population in the absence of CNS disease.  
 
 
The study of Tetrahymena pyroformis growth in different media 
Deana Rodriguez and Emily Ford
Western New England University     
Poster Session A: 1:00-2:15
 
A major cause of cancer in humans is chromosome missegregation during mitosis. Further study of cell cycle transitions, such as mitotic exit, can help to gain insight into how such missegregation can be prevented. The proteins that allow for mitotic exit, the Mitotic Exit Network (MEN) ensure that spindle pole bodies, and attached chromosomes, are properly aligned, which ultimately triggers exit from mitosis back into G1. Studying the MEN in budding yeast may improve our understanding of a homologous, more complex human pathway known as the HIPPO tumor suppressor pathway. Tem1 is a GTPase and an activator of the MEN. Bfa1 acts with Bub2 as a GTPase activating protein that inhibits Tem1. Previous studies have suggested that Bfa1 has an unknown, Bub2-independent role. The overexpression of Bfa1, by the GAL-BFA1 allele, is lethal to cells and impairs Tem1 localization. We are interested in characterizing this GAL-BFA1 allele, and determining if Bfa1 has an unknown secondary function, downstream of its interactions with Bub2 and Tem1. Here we show that GAL-BFA1 does not affect the localization of the downstream MEN proteins, Mob1 and Dbf2, but could be affecting their role in cytokinesis. To further characterize this GAL-BFA1 allele, we examined the activity of a spontaneously identified suppressor, a mutant version of Nud1, NUD1-A308T. Nud1 is a scaffold protein that is phosphorylated, and contains three phosphorylation sites that are critical for the recruitment of downstream MEN proteins. We demonstrated that the NUD1-A308T allele does not suppress the lethal GAL-BFA1 phenotype by re-localizing Tem1. However, NUD1-A308T suppressed otherwise lethal nud1 mutations within its key phosphorylation sites. These results suggest that NUD1-A308T may enhance the localization of downstream components by creating an additional phospho-docking site.
 
 
Assessing Micro-climate Differences Between an Urban College Campus and Neighboring Green Space
Victoria Rosa1, H. Gaughan2, and Lisa Lobel2
1Emmanuel College; 2Wheelock College       
Poster Session B: 4:00-5:15
 
Research shows that urban centers can be up to ten degrees warmer than neighboring rural areas. This phenomenon is known as the urban heat island effect (UHI) and results from urban surfaces like rooftops, roads and parking lots absorbing solar radiation and retaining heat. In comparison, rural areas typically have larger vegetated areas that are cooler due to shading and evaporative cooling. Hotter cities result in increased energy consumption when residents need to turn on their air conditioners. This increased energy consumptions combined with high emissions from vehicles and industry can result in impaired human and environmental health. Green spaces and parks within large scale UHIs have been demonstrated to act as cool islands that can also reduce neighboring urban temperatures. In order to understand micro-scale urban heat island or cool island effects, air temperature and relative humidity were measured at four locations on the Wheelock College campus and three locations in the neighboring Emerald Necklace Park. The park area is a narrow strip of mainly forested parkland, with few paved walking paths that is bisected by the Muddy River. In comparison, Wheelock Campus consists of 5 acres covered with six buildings, paved walkways, patios and parking lots. Air temperature data collected hourly between April 2016 and April 2017 indicate that air temperatures were significantly lower in the Emerald Necklace as compared to Wheelock campus. The green space of the Emerald Necklace had an average annual temperature of 11.3 °C which was 1.2 °C cooler than the average annual temperature of 12.5 °C on Wheelock’s campus.
 
 
The effects of isolation and chemical exposure on Madagascar Hissing Cockroach appearance and behavior
Fiana Saget, Mark Melkun, and Lisa A. E. Kaplan
Quinnipiac University
Poster Session B: 4:00-5:15
 
This pilot study was designed to evaluate exoskeleton appearance (shiny or dull) and leg movement as endpoint indicators of social stress and benzyl butyl phthalate exposure among Madagascar Hissing Cockroaches. Cockroaches were randomly separated into treatment groups: mixed communal (males and females), same-sex communal (males or females), isolated males, isolated females, BBP-exposed males, and BBP-exposed females.  BBP dosage (0.1 mg/L) was administered three times per week for a total of eight weeks. All groups were observed for that same duration. Among male cockroaches, the incidence of shiny exoskeletons decreased 14% (P=0.0001) among those living in a same-sex communal environment and decreased 25% among males housed in isolation (P=0.0001) as compared to mixed communal males (control). Among female cockroaches, however, the incidence of shiny exoskeletons increased 30% for females living in a same-sex communal environment (P=0.0001) and increased 10% among those living isolation (P=0.001) as compared to mixed communal females (control). BBP exposure appeared to increase the incidence of shiny exoskeletons in males by 30% (P=0.0001) as compared to the isolated male control, and have the opposite effect on females where BBP decreased the incidence of shiny exoskeletons by 10% (P=0.0001). Over the eight-week experimental period, no reduction in leg movement was observed among same-sex communal males or same-sex communal females as compared to their mixed communal counterparts. Among isolated males and isolated females, however, leg movement was reduced by more than half (67% and 61%, respectively) as compared to their control mixed communal counterparts (Pmale=0.003; Pfemale=0.006). BBP exposure did not increase leg movement in either males and females as compared to the isolated groups (Pmales=0.17; Pfemales=0.61). Overall, these results indicate that exoskeleton appearance and leg movement are suitable endpoints for the evaluation of social unit disruption and the impact of chemical (BBP) insult. They provide a mechanism for the relatively rapid and harmless assessment of very different sorts of environmental stress.
 
 
The Implications of Dietary Fructose in the Onset of Chemical Colitis
Ariana Savage
Simmons College
3:30–SMT 315
 
Inflammatory Bowel Disease (IBD) is a condition of chronic bowel inflammation which may be caused by genetics, environment, microbiome, and immune system. This study investigates the contribution of high fructose diet on colitis using a chemical model of colitis in male mice. Forty C57BL/6 wild-type male mice were placed into four experimental conditions with ten mice per condition: DSS + fructose, DSS + chow, water + fructose, and water + chow. Dextran Sodium Sulfate (DSS) was used to induce chemical colitis in these animals, and their weight and clinical colitis symptoms (percentage weight loss, appearance, diarrhea, and blood in stool), were evaluated daily. After euthanasia, RNA extraction and reverse transcription were conducted on colon tissues. Quantitative real time PCR was used to examine expression of inflammatory and tight junction genes. Colon histology was assessed for presence of pathology (edema, inflammation, and loss of crypts). Data reveal that mice consuming DSS plus fructose have significantly worse colitis than those consuming DSS alone. We hypothesize that DSS treatment will affect inflammatory and tight junction gene expression and are assessing this now. This model therefore demonstrates that a fructose diet can exacerbate a mouse model of clinical colitis, which is important in clarifying environmental contributors to this disease in humans.
 
 
Characterizing Phage Transduction Between Different Borrelia Genospecies That Cause Lyme Disease
Irio Schiano, Megan Ahern, Maria R. Becker, and Christian H. Eggers        
Quinnipiac University
Poster Session A: 1:00-2:15
 
Pathogens evolve to overcome the challenging environments of a host by using horizontal gene transfer (HGT) to acquire new genes or modify the existing genes necessary to successfully survive and propagate within those environments. Several infection-associated alleles within Borrelia burgdorferi, one of the causative agents of Lyme disease, are known to be subject to HGT. Previous studies have shown that bacteriophage-mediated DNA movement is the likeliest, if not only means of HGT in B. burgdorferi.  While B. burgdorferi is known to cause the vast majority of Lyme disease in the United States, a number of closely-related genospecies cause similar diseases worldwide. In parts of Europe and Asia, for instance, B. burgdorferi, Borrelia afzelii, and Borrelia garinii are all found within the same ecological niches. What is unclear, is whether these genospecies are genetically-isolated or whether HGT via bacteriophage may occur among these genospecies. In this experiment we investigated whether Borrelia burgdorferi can use its bacteriophage to transduce DNA to B. afzelii and B. garinii. To do this, we utilized a recently-published method of co-culture that relies on the use of multiple antibiotic-resistance cassettes to detect DNA movement under phage-inducing conditions. Our preliminary results were unable to detect phage-mediated DNA movement from B. burgdorferi into either B. afzelii or B. garinii; however, we were able to detect DNA movement from these two genospecies into B. burgdorferi by an as-yet-uncharacterized mechanism. This represents the first direct observation of DNA movement between different genospecies of the Lyme disease-causing Borrelia species. Understanding how HGT occurs across genospecies may ultimately allow us to better understand how these bacteria evolve to infect their mammalian hosts, including humans.
 
 
Is coping with novel environments a secondary sex behavioral characteristic in the zebrafish (Danio rerio)? 
Olivia Smiaroski and Maria E. Abate
Simmons College
Poster Session A: 1:00-2:15

    The zebrafish (Danio rerio) is a widely used model organism for studying animal stress responses as well as human anxiety. Strategies to cope with a novel or threatening environment can be categorized into two broad categories: proactive risk-takers are dominant, bold individuals that actively explore the environment, versus cautious, risk-averse individuals. Studies on mammals, birds and fishes suggest that males and females employ opposite coping strategies; and males are often more proactive risk-takers. Genetic strain and husbandry conditions of fishes may also have a role in influencing coping strategies. This study tested whether zebrafish males react more proactively to a novel, stress-inducing environment than females do. Zebrafish (n = 10) were housed in unisex social groups (n = 5). Individual responses to the novel tank test (3 L) were videoed for 10 minutes; and paired t-tests on mean behaviors were performed. On average males from each social group took longer to settle to the bottom of the tank and enter the top half (P < 0.01) which indicates that males are more risk-averse than females. A second experiment explored whether housing fish in unisex groups affected this outcome. Results from novel tank tests of individuals housed in social groups of equal sex ratio will be compared to those from unisex groups. These behavioral findings offer a platform for our future studies regarding the impact of sex hormones on coping strategies. 
 
                                                             
 
You Wnt some you lose some: Wnt/β-catenin and FGF signaling in the posterior lateral line development of Poecilia reticulata        
Alison Smith and Bronwyn H. Bleakley
Stonehill College
2:45–SMT 316
 
The lateral line organ is a mechanosensory organ found in aquatic vertebrates that detects and interprets pressure signals in aquatic environments. The lateral line is comprised of individual neuromasts and the timing of development of the lateral line likely influences the survival of juvenile fish. Development of the lateral line has been best characterized in zebrafish (Danio rerio), where it depends on the action of two cellular migration pathways: Wnt/β-catenin and FGF signaling. Trinidadian guppies (Poecilia reticulata) are a model organism for studying behavioral and life history evolution. Much less is known about the development of their lateral line because they have long duration internal development. This study sought to 1) document lateral line development of guppies for seven days post-parturition, 2) determine if the expression levels of the Wnt/β-catenin via LEF-1 and/or the expression levels of the FGF signaling pathway via FGFR1 change over that time, and 3) determine if differences in the expression of either gene influenced the phenotypic outcome of lateral line development. The lateral line organ was visualized using a fluorescent dye and neuromasts were counted for 158 newborn guppies. Half of the fish were experimentally induced for early birth. Expression levels of LEF-1 and FGFR1 were measured using end-point PCR for control fish across seven post-parturition days. The number of neuromasts changed significantly on the head and occipital region of the fish, although this pattern was driven in large part by the early parturition guppies. Levels of expression of both LEF-1 and FGFR-1 were consistent across development but varied among individuals. Only expression of LEF-1 influenced the number of neuromasts present in the lateral dorsal region. These results suggest that the lateral line organ in guppies develops via the same pathway as zebrafish but is largely complete at birth due to extended developmental time. 
 
 
Investigating the effects of mutant BFA1 and NUD1 alleles on effectors of the Mitotic Exit Network (MEN) in S. cerevisiae
Courtney Sniffen and Anupama Seshan
Emmanuel College
Poster Session A: 1:00-2:15
 
A major cause of cancer in humans is chromosome missegregation during mitosis. Further study of cell cycle transitions, such as mitotic exit, can help to gain insight into how such missegregation can be prevented. The proteins that allow for mitotic exit, the Mitotic Exit Network (MEN) ensure that spindle pole bodies, and attached chromosomes, are properly aligned, which ultimately triggers exit from mitosis back into G1. Studying the MEN in budding yeast may improve our understanding of a homologous, more complex human pathway known as the HIPPO tumor suppressor pathway. Tem1 is a GTPase and an activator of the MEN. Bfa1 acts with Bub2 as a GTPase activating protein that inhibits Tem1. Previous studies have suggested that Bfa1 has an unknown, Bub2-independent role. The overexpression of Bfa1, by the GAL-BFA1 allele, is lethal to cells and impairs Tem1 localization. We are interested in characterizing this GAL-BFA1 allele, and determining if Bfa1 has an unknown secondary function, downstream of its interactions with Bub2 and Tem1. Here we show that GAL-BFA1 does not affect the localization of the downstream MEN proteins, Mob1 and Dbf2, but could be affecting their role in cytokinesis. To further characterize this GAL-BFA1 allele, we examined the activity of a spontaneously identified suppressor, a mutant version of Nud1, NUD1-A308T. Nud1 is a scaffold protein that is phosphorylated, and contains three phosphorylation sites that are critical for the recruitment of downstream MEN proteins. We demonstrated that the NUD1-A308T allele does not suppress the lethal GAL-BFA1 phenotype by re-localizing Tem1. However, NUD1-A308T suppressed otherwise lethal nud1 mutations within its key phosphorylation sites. These results suggest that NUD1-A308T may enhance the localization of downstream components by creating an additional phospho-docking site.
 
 
Dehydration effects on sugar composition in amphibian skin
Daniel Suarez, Y. Madani, Halle Burns, N. Shearer, and Ellen Faszewski
Wheelock College
Poster Session B: 4:00-5:15
 
Previous studies have suggested the importance of carbohydrates in the regulation of water balance in amphibians. For example, the accumulation of sugars, in response to dehydration has been observed in various internal organs of both freeze-tolerant and intolerant species. To further study this relationship, we examined the effects of dehydration on one of the most important regulatory organs, the skin. More specifically, we wanted to understand the role of carbohydrates in the skin during this process.  To do this, dorsal and ventral skin from representative larval stages of the leopard frog, Rana pipiens, were prepared at 0%, 6% and 12% dehydration of body weight. To determine the localization of three sugars, N-Acetyl-Galactosamine (GalNAc), α-L-fucose (Fuc), and N-Acetyl-Glucosamine (GlcNAc), cross sections were then stained with soybean agglutinin (SBA), Ulex europaeus agglutinin (UEA), and wheat germ agglutinin (WGA), respectively. Two negative controls were used: 1) phosphate buffered saline (PBS), and 2) the pre-incubation of each lectin with its inhibitory sugar. Results indicate that lectin staining patterns vary based on lectin, developmental stage, and skin component (epithelial, glands, etc.). This study contributes to our understanding of the role of carbohydrates and their relationship to skin structure and function with a specific emphasis on the regulation of water balance.
 
 
Modeling synapses using patient-derived induced pluripotent stem cells (iPSCs) 
Jessica Thanos1, C. Sellgren2, J. Gracias2, T. Fu2, S. Sheridan2, and R. Perlis2         
1Emmanuel College; 2Massachusetts General Hospital           
Poster Session A: 1:00-2:15
 
Schizophrenia (SZ) is a chronic and debilitating mental illness affecting 1% of individuals. SZ patients experience a variety of severe clinical features, such as positive (e.g. hallucinations and delusions) and negative (e.g. reduced sociability) symptoms as well as cognitive deficits (e.g. impaired memory). Recent studies suggest that abnormal synaptic pruning (i.e. selective synapse elimination) by microglia may be a risk factor for SZ. Complement component 4 protein (C4) has been shown to mediate synaptic pruning, and found to be dysregulated in SZ. However, current studies do not fully capture unique human aspects of SZ. To develop a model of human SZ synapses for future synaptic pruning studies, patient-derived neurons were used to isolate synapses into membrane vesicles. The purity of isolated membrane-bound synapses were assessed by transmission electron microscopy (TEM), and indicated successful isolation and purification of synapses. Next, immunofluorescence was used to assess expression of common neuronal lineage marker proteins, namely beta-tubulin 3 (TUJ1) and microtubule-associated protein 2 (MAP2). Both TUJ1 and MAP2 were expressed by the cultured proteins. Finally, SZ and control synapses were separated by SDS-PAGE and analyzed by western blot. C4 was expressed in SZ synapses because the protein band was approximately the expected size of 200 kDa. Furthermore, C4 expression was higher in SZ synapses as the signal for the SZ protein band was almost twice as intense as that of healthy controls. Future studies will generate more model synapses for SZ and control subjects, and use them for synaptic pruning studies.
 
 
Physical activity is associated with adult bone microarchitecture  
Victoria Turkington1, Kristin L. Popp1,3, Adriana Martinez-Betancourt1, Matthew Scott1, Charlotte Russell2, and Mary L. Bouxsein1,3,4
1Massachusetts General Hospital, 2Simmons College; 3Harvard Medical School; 4Beth Israel Deaconess Medical Center
2:45–SMT 317
 
Stress fracture incidence among United States Military recruits is significantly higher than in the general population. Despite evidence that physically active recruits have a decreased risk of stress fracture throughout basic training, few attempts to estimate bone parameters by quantifying physical activity participation have been made. PURPOSE: This research determined the ability of a Skeletal Loading (SkL) algorithm to predict bone microarchitecture and strength in young adults. METHODS: In this cross- sectional study, 185 young (23.7 ±3.32 years [mean±SD]) White and Black males and females were recruited. Cortical (Ct) and trabecular (Tb) volumetric bone density (vBMD), microarchitecture, and estimated bone strength (by micro-finite element analysis (μFEA)) were assessed at the distal tibia (4% of tibial length) using high-resolution peripheral quantitative computed tomography (82 μm3 voxel size). Physical activity questionnaires were administered and a SkL score derived based on the estimated ground reaction forces, loading rate, frequency, and life-period of participation. Multiple linear regression models determined associations between SkL score and bone outcomes, after adjusting for age, weight, height, sex, and race. RESULTS: The SkL score was significantly associated with Tb area and Ct area, volume, and perimeter measurements. μFEA analysis revealed that predicted stiffness and failure load, as strength parameters, were also associated with the SkL score (all r2 (adjusted) ≥ 0.50, p <0.05). Additional parameters including Ct porosity, thickness, and density were predicted by the SkL score (all r2 (adjusted) = 0.3-0.5, p < 0.05). CONCLUSION: After adjusting for co-variates, the SkL score can predict a significant amount of variability in bone health and strength measurements in young people. Stress fracture prevention methods throughout basic training may be tailored according to the estimated bone strength, architecture, and geometry of recruits.
 
 
Characterization of Xenopus laevis CD4, Using IL-16 as the CD4 Ligand
Taylor Uccello and G. Maniero
Stonehill College
3:15–SMT 316
 
The African Clawed Frog, Xenopus laevis, is a well-characterized model for studying the adaptive vertebrate immune response (Gantress et al., 2002). Although Xenopus have been shown to demonstrate activity consistent with helper T lymphocytes (TH cells), the existence of these cells has not yet been proven (Chida et al., 2011). TH cells are identified by the presence of a cell surface coreceptor known as CD4, which helps to stabilize the interaction of the TH cells with MHC class II molecules on the surface of antigen presenting cells (Murphy, 2012).
Another important, and perhaps ancestral, role of CD4 is as a receptor for the cytokine IL-16. Cytokines are molecules that affect the activity of immune cells. IL-16 binds to CD4 and influences the TH cells in several ways (Center et al., 1996). IL-16 affects TH cells by binding to the CD4 molecule, stimulating TH cell growth and migration (Liu et al., 2007), recruiting cells to the site of inflammation (Center et al., 1996), up-regulating the surface molecule MHC class II on the surface of the T cells (Male et al., 2006), and inhibiting the TH cells ability to respond to allogeneic cells (Theodore et al., 1996).
This research seeks to provide evidence for the existence of the CD4 TH cells in X. laevis through observation of sequence alignments and PCR amplification. Additionally, the responses of the IL-16: CD4 interaction were examined.
 
 
Functionality of EF-G-GFP Fusions in E. coli
Sara VanNest and Paul March
Emmanuel College
Poster Session A: 1:00-2:15
 
The purpose of this experiment is to create a functional green fluorescent protein (GFP) fusion with Elongation Factor G (EF-G) in order to use single cell imaging. EF-G is a protein responsible for translocation during the elongation step of protein synthesis. Translocation occurs in the ribosome when the tRNAs move from the P and A sites into the E and P sites respectively. This includes the movement of the mRNA to correspond with the tRNAs. The dynamics of the ribosome contributes to this movement through inter- and intra- ribosomal subunit rotations but it is unknown exactly how EF-G interreacts with the ribosome more specifically. To determine more specific interactions between EF-G and the ribosome, EF-G is being fused to GFP through random transposition reactions. This will allow EF-G to be visualized under UV light and later be used for single molecule imaging in vivo. This would require a fully functional EF-G-GFP fusion which can be assayed using a temperature sensitive strain of E. coli, PEM102. This strain can grow at 30˚C but not at 42˚C due to a mutation found in EF-G. A plasmid containing wild type EF-G can rescue this sensitivity, so EF-G-GFP fusions were investigated to determine if they also rescue the temperature sensitivity when transformed into PEM102. Twelve candidates were identified, seven of which were unique. No fully functional fusions were identified from the seven unique candidates; they could not completely rescue the PEM102 temperature sensitivity. There are partially functional fusions that have been identified that can give insight into other pathways for producing a fully functional fusion candidate. Further research will lead to new fusions to be assayed for functionality, which can then be used for single molecule imagining for more insight into EF-G and its interactions with the ribosome.
 
 
Investigating the Effects of Small Cardioactive Peptide on the Esophagus of the Gastropod Melibe leonina
Colleen Wallace and James Newcomb
New England College
Poster Session A: 1:00-2:15
 
            Small cardioactive peptide (SCP) is a neurotransmitter that plays a role in digestion in a number of gastropods. In this study, we investigated the effects of three concentrations of SCP , and  on the esophagus of the nudibranch Melibe leonina. The average number of contractions and the frequency of these contractions were recorded for fifteen-minute intervals before, during, and after perfusion with SCP. Statistical analyses of the data with ANOVAs indicated that SCP did not have an effect on either the number or frequency of peristaltic contractions in Melibe. Additional experiments are planned to further test the effects of SCP in Melibe feeding.
 
 
You are terminated: Premature transcription stoppage regulates yeast gene expression following DNA damage
Courtney Whalen1, Claire Moore2, and Jason Kuehner1
1Emmanuel College; 2Tufts University School of Medicine
Poster Session B: 4:00-5:15
 
Gene regulation provides organisms with the ability to utilize energy efficiently and adapt to changes within the organism or its environment. The control of gene expression is essential to cell survival during genotoxic stress, such as DNA damage from ultraviolet (UV) light. Gene expression can be regulated by various mechanisms including pre-mature transcription termination (attenuation). In eukaryotes, transcription attenuation prevents mRNA synthesis, resulting in production of short non-coding RNAs and release of the transcribing polymerase. Attenuation typically occurs in a promoter-proximal region, which favors use of the Nrd1-Nab3- Sen1 (NNS)-dependent termination pathway, while termination in a promoter-distal region favors use of termination pathway coupled with mRNA polyadenylation (pA). Very few attenuators have been characterized with regard to their recognition, and the role of attenuator- based gene regulation during cell stress is unknown. We have identified an attenuator within the yeast DNA repair gene, DEF1, that regulates its expression. We hypothesize that the DEF1 attenuator is a hybrid of both NNS- and pA-dependent terminators, requiring factors from both pathways for efficient recognition. We designed a reporter plasmid that linked DEF1 attenuator function with lacZ expression, allowing us to screen trans-acting mutants that interfered with attenuation. Consistent with our hypothesis, we identified trans-acting mutants in both NNS- and pA-dependent termination factors that disrupted attenuator recognition. Overall, the DEF1 terminator behaved more similarly to a hybrid NNS/pA CYC1 terminator than the strictly NNS- dependent SNR13 terminator control. We confirmed that trans-acting mutants shifted production of attenuated RNA to full-length mRNA via RT-PCR. We also determined the putative RNA- binding site for the Hrp1 termination factor via allele-specific suppression of cis-acting attenuator mutants. Our future characterization of these attenuator recognition factors may reveal a novel regulatory mechanism through which eukaryotic cells alter gene expression in response to DNA damage stress.
 
 
Fluctuation in population densities of intertidal Littorine snails in Cobscook Bay, Maine from 2000-2016          
Alice A. Wynn and Carl L. Merrill
Suffolk University
Poster Session A: 1:00-2:15
 
Population densities (number per m2) of three different species of Atlantic periwinkles, Littorina littorea, Littorina obtusata, and Littorina saxatilus, as well as two potential predators of these snails, dog whelks (Thais lapillus) and green crabs (Carcinus maenus) were measured at three locations of the Cobscook Bay region of Maine over a 16-year period as part of a long-term intertidal survey project. The intertidal sites were: 1) at the shore of the Friedman Field Station in Edmunds TWP, 2) at Reversing Falls Park in Pembroke, and 3) at West Quoddy Head State Park in Lubec.
Abundance data were recorded by various student groups resident at the Freidman Field Station in Maine during the summer months ranging from 2000 to 2016. Depending on number and training of participants, one, occasionally two, and rarely three m2 quadrats were sampled on two or more dates each summer.  Densities were determined at vertical positions randomly chosen each date along a transect of each site during these summers. All members of each of these species found to be present within each quadrat were counted and in some instances measured. The present report focuses on densities of two of these periwinkles in particular, Littorina littorea and L. obtusata, one (L. littorea) because it is heavily harvested by humans and the second (L. obtusata) because it is a potentially important prey item for several shore birds and its density appears to have been in decline. 
Elevation of each vertical position relative to mean low tide (MLW) was determined by vertical profiling using the high or low tide position for each tide sampled as a reference, measuring the change in elevation from that reference to each position, and correcting for lunar cycle tidal oscillations (e.g. spring or neap tides) using correction factors from historical tide tables available from Mobile Geographics (tides.mobilegeographics.com).  Densities of these two species of periwinkles at each location have varied substantially from year to year. 
 
 
Maternal injury in Manduca sexta demonstrates transgenerational influence on embryonic phenotype and gene expression  
Muizz Zaman1, Stephen Winston1, Joe Murray1,2, Joon Hong1, Julia Barnhart1, Wendy Smith1, and Rebecca Rosengaus1
1Northeastern University; 2Le Moyne College
Poster Session B: 4:00-5:15
 
Transgenerational-immunity (TGI) (immunity passed to offspring) has been studied previously in animal models. Progeny of parents exposed to environmental or pathogenic stressors are more resilient when they are exposed to those same stressors. Though this phenomenon is widely reported, the mechanism by which it occurs is still unclear. Prior studies with the tobacco hornworm, Manduca sexta, show when mothers are injected with bacteria, Serratia marcescens, their offspring have an increased variability in embryonic size/weight and enhanced bacterial defenses. Our project further characterized the effects of maternal septic and aseptic injury on offspring, focusing on alterations in development and gene expression. M. sexta females were injected just before adult emergence with sterile saline, or with live or heat-killed S. marcescens. Naïve controls received no treatment. After emergence and mating with untreated males, embryos were examined for changes in size, hatching success, gene expression and histone modification. The progeny of females injected with live bacteria and saline exhibited enhanced variability in egg volume along with increased hatching success. Alterations in gene expression, including upregulation of several immune-related genes were also observed, even in embryos whose mothers were subjected solely to aseptic injury. Despite detectable differences in RNA expression of immune-related genes among treated groups, we observed no global alterations in acetylation of specific histone residues. Future studies will elucidate whether epigenetic changes in specific immune-related genes can help to explain observed increases in RNA expression. Our results are consistent with maternal effects that may enhance offspring survival in stressful environments. Our thanks to support from an NSF REU Site Award 1460628, and from Northeastern University in the form of a Tier 1 Award and Undergraduate Research and Creative Endeavors Awards.