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In Saccharomyces cerevisiae, the protein Bud3 is important in bud site selection. It localizes to the mother-bud neck, where it promotes the axial pattern of budding, wherein a new bud forms next to the bud scar left by the previous cell division. It also localizes the cyclin protein Clb2 to the bud neck, which is important for normal cell cycle progression, including mitotic exit and cytokinesis. We studied the effects of Bud3 phosphorylation site mutations on bud site selection and cell cycle progression. Our study included both alanine mutants, which are nonphosphorylatable, and aspartate/glutamate mutants, which constitutively mimic phosphorylation. To observe bud site selection, we used calcofluor white staining and fluorescence microscopy to visualize bud scars for single, double, and triple mutants. For cell cycle progression, we arrested the cells in G1 using alpha-factor mating pheromone, released them from the arrest, and analyzed budding state over time from 0-180 min using brightfield microscopy. Our results indicate that phosphorylation of Bud3 has a small, but statistically significant effect on bud site selection. The triple alanine mutation combination resulted in decreased axial budding, while aspartate/glutamate mutants showed no significant change in budding pattern. Additionally, we found that phosphorylation does affect cell cycle progression: the triple alanine mutant strain had a delay in budding after release from G1 arrest that persisted through the following cell cycle; in contrast, the triple aspartate/glutamate phosphomimetic mutant strain budded with normal timing and actually progressed more quickly through the cell cycle than the wildtype (WT) strain, possibly indicating division at a smaller-than-normal cell size.
Type 2 diabetes mellitus (T2D) is a chronic metabolic disorder characterized by hyperglycemia and insulin resistance, which the CDC estimates currently afflicts one in ten U.S. adults and is projected to continue to rise sharply over the next 50 years. Acyl peptide enzyme hydrolase (APEH), an enzyme circulating in peripheral blood, has been implicated in rodent models of diabetes. To understand if APEH is affected in diabetes, the APEH activity was measured in the blood serum of patients with T2D compared to age-matched controls. Our results show that APEH activity is 22% lower (p = 0.0163) in T2D patients compared to matched controls. Importantly, there was no observable difference in APEH protein expression between cohorts, suggesting that APEH activity is regulated at the protein level. This work is important as it contributes to our knowledge of T2D and the role of APEH in human disease.
Cathryn L. Carney, Nicholas Antonson, Jose Alvarez, Kaitlynn Foss, Kyle Kettler, Ally Jo Lloyd, Daniel Matzner, Deeksha Mohan, Kara Shroll, Cole Snyder, Mason Van Essen, Joshua W. Vander Windt, Cecelia Miles, Steven L. Matzner
Carnivorous plants are adapted to low nutrient soils and accordingly derive nutrients by trapping animals. Despite being stationary, carnivorous plants are active predators, attracting prey through a combination of visual and olfactory cues. Through course-based undergraduate research projects, we tested the importance of visual cues in the capture of wild type and visually impaired (w1118) Drosophila melanogaster by the carnivorous plants, Drosera capensis and Nepenthes ventricosa. These studies addressed two main questions: 1) Are visual cues important for prey capture by carnivorous plants? (2) Are there differences between the wild type (Ore R) and visually impaired w1118D. melanogaster in their behavioral response to visual cues? Student groups exposed fruit flies to carnivorous plants or artificial traps within 37 L aquariums for 8-10 hours and recorded captures. They discovered that visual cues (particularly within the UV spectrum) can have a significant effect in attracting prey to specific trap types. In addition, visual acuity and irradiance levels can impact capture rates by affecting perception of visual cues. These projects illustrate a unique method for testing hypotheses about the importance of visual cues within the capture process and highlight how course-based research projects can be used to tackle relevant scientific questions.
Luis Jimenez, Matthew Gardner, Joy Bochis, Theranda Jashari, Victoria Ellman, Jenifer Vasquez, Stephanie Zapata, Victorya Ramos, Tina Choe, Mahtab Tazehabadi
A procedure was developed to detect and identify fungi in garden soil, forest soil, and compost samples. Microbial DNA was extracted using the Zymo Microbe DNA MiniPrep protocol. Fungi ribosomal internal transcribed space sequences (ITS) were amplified by PCR using primers ITS1 and ITS4. DNA fragments of approximately 640 base pair were detected in all positive samples. Clone libraries were constructed with amplified DNA by ligating the detected fragments with vector pCR®4-TOPO. Transformations were performed using competent Mix and Go Escherichia coli cells. Plasmids were isolated from each clone using the Zyppy plasmid miniprep protocol and inserts were screened by PCR using M13 DNA primers. Most of the identified fungal species were aligned to the phyla Ascomycota and Basidiomycota. BLAST analysis of clone libraries showed that sequences from compost samples were only comprised of the Ascomycota species Thermomyces lanuginosus. Clones from garden soils were mostly unidentified species closely aligned to Ascomycota and Zygomycota while the most abundant sequences in forest soils were related to the Basidiomycota species Cortinarius flexipes. In garden soil more than 50% of ITS sequences belonged to the Kingdom Plantae with species such as Cardamine hirsute, Stellaria media, and Cerastium dinaricum. However, there were no plant ITS sequences in compost and forest soil. Fungal species belonging to the phylum Ascomycota were widely distributed in the 3 environments studied with the forest soil showing the highest fungal diversity.
The completion of the human genome project led to advances both in technology and information availability. The potential of gene editing is tremendous. In the foreseeable future, scientists and doctors will be capable of repairing defective genes within humans (as is being done in other species) as a means to treat or potentially eliminate specific diseases. There are ethical dilemmas with the use of this technology, including who decides whether someone can or should be treated, and what diseases or traits should be altered. We surveyed undergraduate students from a variety of disciplines about some bioethical issues. With the data collected, we show that, in general, students are in favor of using gene editing technology to help cure disease in adults and children, but less willing to support the use in treating cells prior to fertilization. There is less support for altering non-disease characteristics (e.g. hair color, intelligence, or athletic ability), but male students tend to be more supportive of the use of this technology in this area than female students.
In aquatic environments, many organisms, including bacteria, are affected by polluted runoff. The Jones Falls, a tributary that flows into the Chesapeake Bay, is severely impacted by a variety of problems, including a high amount of surrounding impervious surface and stormwater runoff. Bacteria from the stream were isolated, characterized, and subjected to experiments designed to challenge them in media containing used motor oil. Microbial growth was measured (OD590) after this exposure. It was hypothesized that given that the native bacteria in the Jones Falls were likely previously exposed to used motor oil from urban runoff, some species would favor repeated laboratory treatment with used motor oil. The results of growth curve experiments suggest that some bacterial isolates display hindered growth while others display enhanced growth after exposure to used motor oil. Given that microorganisms are the base of the food web, their responses to a changed environment can potentially affect the rest of the aquatic ecosystem. Learning about the microbial species affected by various components of stormwater runoff can aid in identifying release locations and informing mitigation methods.
This study investigated the bacterial communities in soils of three different managements - lawn managed chemically, vegetable garden following organic practices, and unmanaged forest - on the Furman University campus in South Carolina. We hypothesized that the bacterial community in the organically managed soil would be most diverse and that the three bacterial community compositions would differ at least at the family taxonomic level. We took two approaches: culture-dependent and culture-independent. The culture-dependent approach revealed that the bacterial community in the organically managed soil was the most morphologically diverse and the one in the non-managed soil was the least. The Sorenson's coefficient for community similarity showed low similarity between these three bacterial communities. The culture-independent approach revealed that, based on Simpson's diversity indices comparing the bacterial communities at the family level, the bacterial community in the organically managed soil was the most diverse and the one in the chemically managed soil was the least. Additionally, Sorenson's coefficient for community similarity revealed low community similarity between these three bacterial communities. Results from both approaches suggested that the bacterial communities in soil of different managements differ in both composition and diversity and that the bacterial communities are most diverse in organically managed soil.
Tar balls have been persisting along the Gulf coast. Previous research has shown that tar balls from the Gulf coast may contain the human pathogen Vibrio vulnificus. These oil residuals are also commonly found beneath sea shells (hereby called “tar shells”). No research, however, has explored the bacterial communities in tar shells. In this study, tar shell, tar ball, and sand samples on the Alabama Gulf coast at intertidal and supratidal zones were collected and their bacterial communities were assessed using PCR-DGGE and DNA sequencing. Our data suggest that Desulfovibrio and Actinobacteria spp. are dominant in tar shells at intertidal and supratidal zones, but not in tar balls at supratidal zone, while Pseudoalteromonas spp. are dominant in both tar balls and tar shells. This research provides preliminary data for future studies to assess the impacts of tar balls and tar shells on coastal environments and human health.
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