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THE EFFECT OF CANNABINOIDS ON THE DEVELOPMENTAL MORPHOLOGY OF ZEBRAFISH: A SUMMARY**
No full textResults from the past few years on exposure of zebrafish embryos to varying concentrations of delta nine-tetrahydrocannabinol (Δ9-THC), Δ8-THC and cannabidiol demonstrated delayed hatch rate, higher death rate, lowered heart rates and shorter embryos with curved spines when compared to control embryos. We also observed alterations on morphology of jaw structures though have yet to see a consistent pattern. In this study we are processing embryos from last year stored in 100% methanol and are using Alcian Blue staining to examine jaw structure in an effort to identify a consistent pattern of abnormality. We will also analyze data across all three years to summarize overarching trends
APPLICATION OF CRISPR TO REMOVE MULTIPLE BETA-LACTAMASE GENE VARIANTS**
No full textIn modern healthcare settings, antibiotic resistant bacteria are increasingly becoming an issue. Bacteria are becoming resistant at a faster rate than we can develop drugs for, which has lead to alternatives being sought out. A solution could be to remove resistance using gene editing technology like CRISPR. This study focused on β-lactamases specifically, which confer resistance to some of the most common drugs. These enzymes work by hydrolyzing the β-lactam ring in penicillin and cephalosporin class antibiotics, and are divided into four classes based on protein sequence. It can be assumed that the active site of β-lactamases within each class are similar, supported by the fact that 3 of the 4 classes all use a serine amino acid as the core component of the active site. By applying CRISPR-Cas9, this paper examines the potential to remove multiple gene variants using a single guide RNA. The plasmids pHflu2 and pETcon-NK were used, which contain the genes for β-lactamases TEM-116 and TEM-1 and chloramphenicol and kanamycin resistance, respectively. TEM-116 and TEM-1 are variants of the TEM gene, which is a class A β-lactamase. These plasmids were inserted via electroporation into the Escherichia coli strain LC-EO3, which contains a Cas9 gene, to generate two cultures of LC-EO3 each containing one of the β-lactamase plasmids. A plasmid containing a guide RNA that targets TEM-116 was synthesized by Sigma Aldrich and called pAmpR (SRID). Plasmid pAmpR (SRID) also contains an ampicillin resistance cassette. The LC-EO3 strains containing pHflu2 and pETcon-NK will be transformed by electroporation with pAmpR (SRID). These strains will then be replica plated on Luria agar containing ampicillin, chloramphenicol or kanamycin. It is expected that the Cas9, guided by one sgRNA will cause double strand breaks in pHflu2 and potentially pETcon-NK, causing the bacteria to become sensitive to the antibiotics chloramphenicol and kanamycin
THE ROLE OF ADENOVIRUS PROTEIN E4 11K IN P BODY PROTEIN RELOCALIZATION**
No full textAdenoviruses (Ad) can cause diseases, such as the common cold and conjunctivitis. E4 11k is an important viral protein that regulates late gene expression. E4 11k has been shown to disrupt cellular function by relocalizing processing body (p body) proteins to cytoplasmic aggresomes. Aggresomes are sites of misfolded protein storage and only arise when there is cellular stress. The scaffolding p body protein, Ddx6, has been shown to colocalize with E4 11k in aggresomes during a wild-type Ad infection. Ddx6, however, was not relocalized to cadmium chloride (CdCl2)-induced aggresomes. This suggests that E4 11k is necessary for the relocalization of Ddx6. We observed the localization of additional p body proteins (Lsm1, Edc3, and Pat1b) in human lung (A549) cells following wild-type Ad infection and CdCl2 treatment. Lsm1 and Edc3 were relocalized to both infection- and chemically-induced aggresomes. Pat1b, however, was not relocalized to either infection or chemically induced aggresomes. Additionally, we infected A549 cells with three E4 11k-expressing viruses: E4 11k (wild-type), L103A mutant, and D105A/L106A (DL) mutant viruses. The L103A mutant cannot oligomerize but has the ability to dimerize, while the DL mutant virus displays some form of multimerization. Current observations indicate that E4 11k is necessary and sufficient to relocalize Ddx6 to the aggresome and that the level of protein folding present may affect certain functions of p body relocalization. Using these mutant viruses, we hope to determine if the oligomerization from E4 11k is required for aggresome formation. In addition, we want to observe the different p body proteins to determine if their localization during mutant virus infection is altered. We hope to better understand the dynamics of p body and aggresome formation in human cells
MASTER COURSE TEMPLATE IMPLEMENTATION EFFECT OF DFW RATES AND STUDENT SUCCESS IN ONLINE INTRODUCTORY ASTRONOMY **
No full textA study of the effect of the implementation of a collaboratively developed Master Online Course (MOC) in an online introductory astronomy course on overall student course grades shows improvements for the principal investigator’s sections but not for others at the same campus with low enrollment sections. However, concurrent MOC implementation with the inception of an online introductory astronomy course at a sister campus with large enrollment sections yields higher overall student course grades than face-to-face sections on the same campus. An analysis of subsequent targeted data pulls will reveal indications of potential explanations and how to improve student success
WHAT IS REASONABLE? EFFECTS OF DIFFERENT JURY INSTRUCTIONS IN CASES OF POLICE ABUSE OF FORCE**
No full textAccording to the Graham v Conner Supreme Court case, police officers charged with an on-duty crime should be judged against the standard of a reasonable officer. However, the Supreme Court gave an unclear and subjective definition of a reasonable officer, only mentioning that an officer should not be judged with the 20/20 vision of hindsight and that jurors should consider the split-second judgments that an officer in the moment would have. Multiple cases cite Graham v Connor. Federal Law Enforcement Training Center’s Legal Division has a nine-part podcast on how the wording of Graham v Connor can be used to keep officers from going to trial or being convicted. However, some states go beyond the wording of Graham v Connor. For example, California included the totality of circumstances in their pattern jury instructions. The California instructions did not mention the concept of looking at police misconduct without the 20/20 vision of hindsight. My research seeks to determine the effects of different jury instructions in lethal police misconduct cases. I will also look for interactions between juror instructions and juror political affiliation, and Social Dominance Orientation scores. My research uses two types of jury instructions: one set based on California pattern jury instructions, which mention the concept of the totality of circumstances, and juror instructions based on the exact wording of Graham v Connor. I hypothesized that participants given the pattern juror instructions from California will be more likely to convict the police officer than participants given the Graham v Connor jury instructions
Changes in Morphology, Phenology, and Demography of Several Avian Species Over 18 Years at a Restored Grassland
Get PDFIn response to a changing climate and the loss of habitat, avian species are displaying changes in body size, migration phenology, and demography. Several species have adhered to Allen’s and Bergmann’s rules by displaying increased wing length and decreased mass, respectively, over time in response to higher temperatures. Climate change has also resulted in earlier spring migration and depending on species, either earlier or later fall migration. Finally, species affected by habitat loss often display male-skewed sex ratios and juvenile skewed age ratios. Our goal was to determine how body size, migration phenology, and demography have changed for grassland birds over 17 years. We used data from birds captured in mist-nets between 2007 and 2024 at a restored grassland, and fit generalized linear models (body size and migration phenology) and linear regressions (age and sex ratios). Contrary to Allen and Bergmann’s rules, three species displayed shorter wings and two showed greater fat-free mass over time; this is likely in response to changes in climate variability, rather than increasing mean temperatures, or it is unrelated to climate change. One species, unexpectedly, exhibited later spring migration, possibly due to longer migration distances or increased severe weather events during migration, and two species exhibited later fall migration, consistent with other literature. One species displayed a male-skewed sex ratio, possibly a result of increased predation at Panola Mountain State Park or an indication of a declining capture rate for that species. One species showed a juvenile-skewed age ratio, likely due to increased fecundity resulting from improved habitat, or mist netting resulted in a biased sample
A survey of Fusarium oxysporum in Georgia hemp farms and vegetable gardens
Get PDFRecent changes in U.S. federal law have recognized the genetic and chemical differences between hemp and marijuana (Cannabis sativa L.), resulting in the legalization of hemp in the United States. The barriers to this plant’s growth in the state of Georgia are not well understood due to its prohibition for the past 80 years. Among the many possible obstacles is fungal disease. Fusarium oxysporum (Schlecht. emend. Snyder & Hansen) is a fungus that damages various crops and was previously researched as a biocontrol agent for C. sativa. Thus, this study surveyed hemp farms in Georgia to determine the prevalence of F. oxysporum in native soils via qPCR and correlate their abundance with the physicochemical properties of the soil (soil moisture, pH, ammonium, nitrate, and total inorganic nitrogen). Home vegetable gardens were also surveyed to be compared to the F. oxysporum abundance of hemp farms. We found that 5 out of 20 of the hemp soil samples and 1 out of 8 of the vegetable garden samples tested positive for F. oxysporum. Correlation analysis revealed a strong negative correlation between F. oxysporum abundance and soil moisture (r = -0.850, P=0.031). Although there were several plants reported to have symptoms matching that of fusarium wilt, no hemp farmer reported a substantial loss of crops from fungal disease. Overall, the findings of this study suggest that F. oxysporum may not pose a significant barrier to hemp farming in Georgia
LARVAE UNDER PRESSURE: EXPLORING GROUP DYNAMICS AND REGURGITATIVE REPONSES OF TRICHOPLUSIA NI LARVAE**
No full textIn the face of predation, larval insects exhibit diverse defensive responses to enhance survival. However, defensive activities such as unique movement behaviors or the release of compounds combined with unique group dynamics can incur fitness costs to the individual and the group. Therefore, understanding how group dynamics alter the risk-reward behavior of defense is important. For example, in the lepidopteran Pieris brassicae, the group-housing of larvae causes a reduction in defensive action of regurgitation. This study aims to investigate a different lepidopteran species, Trichoplusia ni, with specific focus on larval regurgitation frequency, fitness costs, and variations in defensive behaviors based on grouped (10 larvae housed together from the first in-star) or individual housing. We have a working protocol to elicit regurgitation using a pair of standardized pressure forceps to simulate a predatory attack. We hypothesize that a decrease in group-housed regurgitation rates (regurgitation event per 20 forceps squeezes) compared to single-housed individuals results in a fitness trade-off in T. ni. To this end, we will compare regurgitation rates of individually housed subjects to group-housed subjects and analyze the data using a t-test. Furthermore, we will explore the fitness cost by measuring pupal weight, which we hypothesize to be negatively correlated with regurgitation rate. Additional behavioral data will be collected by coding behaviors observed during and after the simulated attack sequence. We hypothesize unique behavioral responses between regurgitators and non-regurgitators, which may indicate regurgitation is indeed part of a larger behavioral response and not a random event incited by pressure on the gastric cavity. This study contributes to our understanding of defensive behaviors in T. ni and expands the species ethogram whilst shedding light on the role of group dynamics in eliciting defensive behaviors
INVESTIGATING THE ECOSYSTEM ECOLOGY OF SEAGRASS BEDS IN TAMPA BAY, FLORIDA USING BAITED REMOTE UNDERWATER VIDEO (BRUV)**
No full textSeagrass beds are one of the most highly productive ecosystems on the planet and provide many ecosystem services. They are being threatened worldwide by anthropogenic activities which include trawling, urbanization, and eutrophication. In this study we focused on the seagrass ecosystems in Tampa Bay, Florida to understand species’ ecology using baited remote underwater video (BRUV) as non-destructive sampling method. The study’s goal is to identify species richness along with inter- and intra-species interaction within Tampa Bay in relation to the depth, water chemistry, season, proximity to seagrass beds, and proximity to anthropogenic activities. Each month between February 2022 and January 2023, we deployed 15 BRUV units equally divided into a shallow depth (1.8 to 3m) group, a mid-depth (3 to 4.8m) group, and a deep depth (4.8 to 6.7 m) group. All three groups paralleled a shallow seagrass bed as well as a deep boating channel and were deployed in a latitudinal gradient. Extensive sampling with 15 BRUVS per depth group was done in February and October 2022. At each deployment site, we collected water quality including turbidity, pH, nitrite, phosphate, salinity, and up to 1.5 hours of video footage. Preliminary results indicate species richness differed among depth groups, with 11 species observed at shallow depths, 13 species observed at mid-depths, and 16 different species observed at deep depths. Identified species varied with depth and species were found across all three depths with the most common being the Hardhead Catfish (Ariopsis felis). Ongoing research is being conducted to identify the inter- and intra-species interactions. We will then test species richness and interactions for any environmental, temporal, and spatial relationships. With this study, we hope to better understand seagrass ecosystems and surrounding habitats by using non-destructive methods to survey coastal communities and identify mobile organisms
ASSESSING CHATTAHOOCHEE RIVER WATER QUALITY BY MONITORING E. COLI LEVELS
No full textAs the largest river in Georgia, the Chattahoochee provides most of the state’s drinking water and many recreational areas. Since Chattahoochee Riverkeeper was founded in 1994, the Neighborhood Water Watch (NWW) program has been monitoring approximately 400 sites along the river and its tributaries, and Escherichia coli (E. coli) levels have been used as a primary assessment tool for water contamination. In this presentation, two representative cases are discussed to demonstrate the importance of the NWW monitoring system for local water quality: 1. Sewage spill: a leakage from Big Creek Wastewater Treatment Plant was found near Willeo Park by late June 2023. Water samples collected from multiple sites downstream showed very high E. coli levels. Part of the river was closed for recreation until E. coli levels were back to normal about two weeks later; 2. Headless goats: dead goats have been spotted near the I-20 bridge by Six Flags and downstream since 2018. The carcasses were left rotting on the river, severely contaminating the water, as reported by Charles Bethea in the New Yorker. Samples from the Fairburn Road site continuously show extremely high E. coli levels. More research is needed to find the source and purpose of these goats, as well as the pollution they bring to the river