Bird Abstracts
James Bautista – The Effect of Fructose-1,6-bisphosphatases in Autophagy Upregulation
Coxiella burnetii is an intracellular pathogen that causes the zoonotic disease, Q fever. During infection, C. burnetii manipulates the host cell into forming an environment conducive to its replication. It does this by constructing a lysosome-derived vacuole where the pathogen resides and replicates, termed the Coxiella-containing vacuole (CCV). As the CCV expands, it fuses with mature autophagosomes resulting in the surface accumulation of microtubule-associated protein 1A/1B light chain 3-B (LC3-B). Concurrently, C. burnetii injects 130+ effector proteins into host cytosol through its Dot/Icm type IVB secretion system (T4SS) that facilitate CCV biogenesis. Among those, CinF is released. Specifically, cinF encodes for a protein annotated as a fructose-1,6-bisphosphatase (F6BP). A transposon mutant of cinF led to the failure of CCV biogenesis with autophagosomes, suggesting CinF upregulates host autophagy. The aim of our research is to understand if F6BP enzymes induce autophagy, or if it is intrinsic to CinF alone. To solve this, we performed tests expressing the F6BP gene from Coxiella burnetii and Bradyrhizobium japonicum. Then, we quantified intracellular LC3-B levels via immunoblotting to determine the effect of the fructose-1,6-bisphosphatases. This experiment shows promise in understanding a role of C. burnetii’s effector proteins and provides insight on how cells regulate autophagy.
Dakota Cao – Muller Expansion Project Annotation of contig2 in D. wilistoniThe process of gene annotation is crucial to discovering gene function. Annotation is the process in which identifying features of an unmapped genome are labeled so that the genome may be organized into a manner in which it is able to be processed. We looked at contig2 from Drosophila willistoni alongside Drosophila melanogaster using the Genomics Education Partnership (GEP) protocol coursework, the GEP Genome browser, and the Basic Local Alignment Search Tool (BLAST) to perform exon by exon searches identifying exact start and stop coordinates of potential conserved ancestral genes by locating model coding sequences in the GEP Gene Record finder of the orthologs and performing a BLAST the exons using the NCBI BLASTx tool to attain coding exon start/stop boundaries which we then investigated on the genome browser to determine coordinates that often needed adjusted past homology predictions. The genome tracks used included the genome and translated ORFs in willistoni, the splice site sequence rules, etc. These coordinates were confirmed to the Gene Model checker. The gene structures annotated from contig2 were mRpL55, ATPsynD, CG6026, and cdi. Overall, four gene homologs were annotated successfully, with miniscule shifts in homology.
Joseph Chrisman – Isolation of Protein Isolates from Brewer Spent GrainEvery year, 39 million tons of brewers’ spent grain (BSG) is produced after the brewing of beer; making up roughly 85% of the waste produced. Typically, BSG is incorporated into animal feed due to the low price to ranchers. However, BSG is rich in hemicellulose (20-25%), cellulose (12-25%), ash (2-5%), lipids (10%), lignin (12-28%) and most importantly protein (19-30%), making it ideal for incorporation into the human diet. This study's objective was to identify the isoelectric point of the protein within BSG through pH manipulation to maximize the total protein yield. The isoelectric point is defined as the pH at which the electrostatic charge of a protein is at zero which would cause it to become water insoluble. The Bradford method will be used for protein analysis. The pH treatments with the lowest and greatest yields were 6 and 6.5 that produced an average yield of 3.7137 (±0.6664) and 5.4928 (±0.6664), respectfully. ANOVAs was used to examine significant differences (p<0.05) using Excel. With this as a preliminary study, it will serve as a foundation towards understanding how protein isolated from BSG could be used within the human diet.
Blue Cunningham – Nutritional Preferences of Male and Female Fruit Flies (Drosophila melanogaster)Male and female insects often have different nutritional needs. The eggs require protein and lipids, causing the females to search for food with high concentration of these nutrients. Males only need carbohydrates for energy to find females and mate. We tested the effect of different concentrations (9% vs. 4%) of yeast solution dyed either red or blue on the foraging behavior and feeding preference of fruit flies (Drosophila melanogaster) on a patch of food (yeast solution) in a closed arena. 50 flies were deprived of food for 24 hours and allowed to feed together for 1 hour in the dark. They were then frozen, and their abdomens were evaluated for color to determine their preferred yeast concentration. The feeding patches were square containers (3.4x3.4x0.4cm) with 25 wells containing 10μL of solution. We tested the flies in groups of males only, females only, and mixed sexes. When females fed alone or when both sexes fed together the flies preferred the higher yeast concentration. However, when males fed alone, they showed no preference. This shows that the females have a clear preference for a higher level of nutrients than the males during foraging.
Oanh Dang & Katherine Dempsey – Cytotoxicity of 6-Gingerol on Colorectal Cancer Cell ViabilityColorectal cancer is the third most common type of cancer and the second leading cause of cancer-related deaths in America. Common treatment options include invasive surgeries and chemotherapy but are often accompanied by complications of sepsis, bowel stress, and reduced immunity. Therefore, the search continues for natural therapeutic alternatives that differentiate between healthy and cancerous tissues. Zingiber officinale, commonly known as the ginger root, has been used for centuries in ancient Chinese and Indian Ayurvedic medicine due to its well-known anti-inflammatory and antioxidant properties. It was hypothesized that the main bioactive compound in ginger, 6-gingerol, may possess anticancer properties that would reduce the cell viability of colorectal cancer in a dose-dependent manner. Microscopic observations of HT-29 cells treated with gingerol were consistent with apoptotic morphology. Cell viability and caspase activity assays further supported the theory that gingerol induces apoptosis in cancer cells. In comparison, HEK293T cells displayed increased cell viability when treated with gingerol. Analysis of both qualitative and quantitative data suggest that gingerol is able to selectively target tumorigenic cells while promoting viability in noncancerous cells.
Alexa Gallegos – Evaluating the Effect of Ricinoleic Acid on the Synthesis of Collagen in NIH/3T3 Fibroblast CellsCastor oil has been used in many applications, due to the presence of the major component, ricinoleic acid (RA) and its properties. As a skin-conditioning agent, it has been studied that ricinoleic acid has wound-healing properties, so anecdotally, skin elasticity and rejuvenation has gotten associated with the claim. However, the claims remain unclear as ricinoleic acid has not been studied for collagen, the dermal connective tissue agent for strength, elasticity, and integrity. In this research, the effects of 50 μM and 100 μM ricinoleic acid on collagen synthesis, using collagen and non-collagenous protein concentration were analyzed in a mouse embryonic fibroblast cell line (NIH/3T3) with a Sirius red/fast green (SR/FG) collagen staining assay. Previous work has shown that cells are viable at 50 μM of ricinoleic acid with an MTT assay, but the increased dose of 100 μM in the SRFG collagen staining assay show a decreased in values. The analysis of NIH/3T3 fibroblasts treated with 50 μM and 100 μM ricinoleic acid will determine the effect on growth of collagen synthesis compared to those cells treated without ricinoleic acid.
Katelynn Graves – DNA Barcoding of West Texas InsectsTo examine the diets of insectivorous bat species using DNA sequences, we must have a well-documented genetic database of the prey they consume. The Genbank database does not have a complete record of insects and moths of West Texas, and generating genetic information from these insect species could improve database coverage and provide reference sequences for future studies using a molecular approach. In this study, we sequenced the cytochrome c oxidase subunit I (COI) gene from 20 insects collected from Presidio, Texas. Of the original morphological insect identifications, 35% matched the molecular identifications on GenBank Blast. Of the original identifications, 40% were misidentifications. Only 15% of the molecular identifications hinted at contamination as a result of challenges in eliminating moth scales from some of the insects. Of molecular sequences, 10% were high quality but had less than 95% identity to the database indicating that these taxa may not be represented in Genbank. Overall, the study revealed discrepancies between morphological and molecular identifications, with misidentifications and indications of potential contamination, alongside possible taxa absent in GenBank.
Natalie Ramirez – Cytotoxicity of Hericium erinaceus Mushroom Secretions Against U-87 MG GlioblastomasWith a five-year survival rate of 6.8%, glioblastoma has poor survival outcomes for patients. Current treatments of glioblastoma include surgical removal of the tumor followed by intensive radiation therapy and chemotherapy. The development of a natural remedy with reduced side effects would significantly benefit cancer patients. Hericium erinaceus, better known as Lion’s Mane, has been traditionally used to enhance immune system function, treat gastric ulcers, and aid the body in controlling cancer cell growth. Additionally, studies on H. erinaceus have suggested that it increases cognitive ability and improves brain function. Because of H. erinaceus’ advantageous nervous system functions, this study focuses on the cytotoxic effects of H. erinaceus secretions against U-87 MG glioblastoma cells in vitro. Full spectrum secretions from the mycelia, as well as three combinations of fractions, were used in a cell cytotoxicity assay. Results from the assay suggest that secretions from H. erinaceus may be efficacious in causing U-87 MG cells to undergo apoptosis as compared to nontumorigenic MCF10A epithelial cells.
Pritika Shree Thotakura – DNA Barcoding as IdentificationBees play a crucial role in pollinating crops such as almonds and strawberries. However, the success of crop yields not only depends on the number of bees but also on the diversity of bee species in the area. Bee species differ in their behavior and plant preferences, which affects their pollination success. Therefore, it is essential to understand the variety of bees in an area to ensure agricultural success. To measure bee diversity, researchers can use a tool called DNA barcoding, which assigns a unique "barcode" to each species based on sequences with high interspecies variation but low intraspecies variation. This allows for highly accurate identification based on DNA sequences alone. By creating a database of these unique DNA sequences, researchers can identify the different species of bees in a specific area accurately. This can help reduce the number of misidentifications that may arise from morphological identification. Combining morphological identifications with barcoding identifications can also help reaffirm the efficacy of using morphology to identify bees. Our study aims to determine the DNA barcodes of bee species collected in Grayson Co., TX, and add them to the Barcode of Life Data Systems (BOLD) database. By contributing to these databases, a more thorough understanding of the local North Texas bee species may be achieved.
Coxiella burnetii is an intracellular pathogen that causes the zoonotic disease, Q fever. During infection, C. burnetii manipulates the host cell into forming an environment conducive to its replication. It does this by constructing a lysosome-derived vacuole where the pathogen resides and replicates, termed the Coxiella-containing vacuole (CCV). As the CCV expands, it fuses with mature autophagosomes resulting in the surface accumulation of microtubule-associated protein 1A/1B light chain 3-B (LC3-B). Concurrently, C. burnetii injects 130+ effector proteins into host cytosol through its Dot/Icm type IVB secretion system (T4SS) that facilitate CCV biogenesis. Among those, CinF is released. Specifically, cinF encodes for a protein annotated as a fructose-1,6-bisphosphatase (F6BP). A transposon mutant of cinF led to the failure of CCV biogenesis with autophagosomes, suggesting CinF upregulates host autophagy. The aim of our research is to understand if F6BP enzymes induce autophagy, or if it is intrinsic to CinF alone. To solve this, we performed tests expressing the F6BP gene from Coxiella burnetii and Bradyrhizobium japonicum. Then, we quantified intracellular LC3-B levels via immunoblotting to determine the effect of the fructose-1,6-bisphosphatases. This experiment shows promise in understanding a role of C. burnetii’s effector proteins and provides insight on how cells regulate autophagy.
Dakota Cao – Muller Expansion Project Annotation of contig2 in D. wilistoniThe process of gene annotation is crucial to discovering gene function. Annotation is the process in which identifying features of an unmapped genome are labeled so that the genome may be organized into a manner in which it is able to be processed. We looked at contig2 from Drosophila willistoni alongside Drosophila melanogaster using the Genomics Education Partnership (GEP) protocol coursework, the GEP Genome browser, and the Basic Local Alignment Search Tool (BLAST) to perform exon by exon searches identifying exact start and stop coordinates of potential conserved ancestral genes by locating model coding sequences in the GEP Gene Record finder of the orthologs and performing a BLAST the exons using the NCBI BLASTx tool to attain coding exon start/stop boundaries which we then investigated on the genome browser to determine coordinates that often needed adjusted past homology predictions. The genome tracks used included the genome and translated ORFs in willistoni, the splice site sequence rules, etc. These coordinates were confirmed to the Gene Model checker. The gene structures annotated from contig2 were mRpL55, ATPsynD, CG6026, and cdi. Overall, four gene homologs were annotated successfully, with miniscule shifts in homology.
Joseph Chrisman – Isolation of Protein Isolates from Brewer Spent GrainEvery year, 39 million tons of brewers’ spent grain (BSG) is produced after the brewing of beer; making up roughly 85% of the waste produced. Typically, BSG is incorporated into animal feed due to the low price to ranchers. However, BSG is rich in hemicellulose (20-25%), cellulose (12-25%), ash (2-5%), lipids (10%), lignin (12-28%) and most importantly protein (19-30%), making it ideal for incorporation into the human diet. This study's objective was to identify the isoelectric point of the protein within BSG through pH manipulation to maximize the total protein yield. The isoelectric point is defined as the pH at which the electrostatic charge of a protein is at zero which would cause it to become water insoluble. The Bradford method will be used for protein analysis. The pH treatments with the lowest and greatest yields were 6 and 6.5 that produced an average yield of 3.7137 (±0.6664) and 5.4928 (±0.6664), respectfully. ANOVAs was used to examine significant differences (p<0.05) using Excel. With this as a preliminary study, it will serve as a foundation towards understanding how protein isolated from BSG could be used within the human diet.
Blue Cunningham – Nutritional Preferences of Male and Female Fruit Flies (Drosophila melanogaster)Male and female insects often have different nutritional needs. The eggs require protein and lipids, causing the females to search for food with high concentration of these nutrients. Males only need carbohydrates for energy to find females and mate. We tested the effect of different concentrations (9% vs. 4%) of yeast solution dyed either red or blue on the foraging behavior and feeding preference of fruit flies (Drosophila melanogaster) on a patch of food (yeast solution) in a closed arena. 50 flies were deprived of food for 24 hours and allowed to feed together for 1 hour in the dark. They were then frozen, and their abdomens were evaluated for color to determine their preferred yeast concentration. The feeding patches were square containers (3.4x3.4x0.4cm) with 25 wells containing 10μL of solution. We tested the flies in groups of males only, females only, and mixed sexes. When females fed alone or when both sexes fed together the flies preferred the higher yeast concentration. However, when males fed alone, they showed no preference. This shows that the females have a clear preference for a higher level of nutrients than the males during foraging.
Oanh Dang & Katherine Dempsey – Cytotoxicity of 6-Gingerol on Colorectal Cancer Cell ViabilityColorectal cancer is the third most common type of cancer and the second leading cause of cancer-related deaths in America. Common treatment options include invasive surgeries and chemotherapy but are often accompanied by complications of sepsis, bowel stress, and reduced immunity. Therefore, the search continues for natural therapeutic alternatives that differentiate between healthy and cancerous tissues. Zingiber officinale, commonly known as the ginger root, has been used for centuries in ancient Chinese and Indian Ayurvedic medicine due to its well-known anti-inflammatory and antioxidant properties. It was hypothesized that the main bioactive compound in ginger, 6-gingerol, may possess anticancer properties that would reduce the cell viability of colorectal cancer in a dose-dependent manner. Microscopic observations of HT-29 cells treated with gingerol were consistent with apoptotic morphology. Cell viability and caspase activity assays further supported the theory that gingerol induces apoptosis in cancer cells. In comparison, HEK293T cells displayed increased cell viability when treated with gingerol. Analysis of both qualitative and quantitative data suggest that gingerol is able to selectively target tumorigenic cells while promoting viability in noncancerous cells.
Alexa Gallegos – Evaluating the Effect of Ricinoleic Acid on the Synthesis of Collagen in NIH/3T3 Fibroblast CellsCastor oil has been used in many applications, due to the presence of the major component, ricinoleic acid (RA) and its properties. As a skin-conditioning agent, it has been studied that ricinoleic acid has wound-healing properties, so anecdotally, skin elasticity and rejuvenation has gotten associated with the claim. However, the claims remain unclear as ricinoleic acid has not been studied for collagen, the dermal connective tissue agent for strength, elasticity, and integrity. In this research, the effects of 50 μM and 100 μM ricinoleic acid on collagen synthesis, using collagen and non-collagenous protein concentration were analyzed in a mouse embryonic fibroblast cell line (NIH/3T3) with a Sirius red/fast green (SR/FG) collagen staining assay. Previous work has shown that cells are viable at 50 μM of ricinoleic acid with an MTT assay, but the increased dose of 100 μM in the SRFG collagen staining assay show a decreased in values. The analysis of NIH/3T3 fibroblasts treated with 50 μM and 100 μM ricinoleic acid will determine the effect on growth of collagen synthesis compared to those cells treated without ricinoleic acid.
Katelynn Graves – DNA Barcoding of West Texas InsectsTo examine the diets of insectivorous bat species using DNA sequences, we must have a well-documented genetic database of the prey they consume. The Genbank database does not have a complete record of insects and moths of West Texas, and generating genetic information from these insect species could improve database coverage and provide reference sequences for future studies using a molecular approach. In this study, we sequenced the cytochrome c oxidase subunit I (COI) gene from 20 insects collected from Presidio, Texas. Of the original morphological insect identifications, 35% matched the molecular identifications on GenBank Blast. Of the original identifications, 40% were misidentifications. Only 15% of the molecular identifications hinted at contamination as a result of challenges in eliminating moth scales from some of the insects. Of molecular sequences, 10% were high quality but had less than 95% identity to the database indicating that these taxa may not be represented in Genbank. Overall, the study revealed discrepancies between morphological and molecular identifications, with misidentifications and indications of potential contamination, alongside possible taxa absent in GenBank.
Natalie Ramirez – Cytotoxicity of Hericium erinaceus Mushroom Secretions Against U-87 MG GlioblastomasWith a five-year survival rate of 6.8%, glioblastoma has poor survival outcomes for patients. Current treatments of glioblastoma include surgical removal of the tumor followed by intensive radiation therapy and chemotherapy. The development of a natural remedy with reduced side effects would significantly benefit cancer patients. Hericium erinaceus, better known as Lion’s Mane, has been traditionally used to enhance immune system function, treat gastric ulcers, and aid the body in controlling cancer cell growth. Additionally, studies on H. erinaceus have suggested that it increases cognitive ability and improves brain function. Because of H. erinaceus’ advantageous nervous system functions, this study focuses on the cytotoxic effects of H. erinaceus secretions against U-87 MG glioblastoma cells in vitro. Full spectrum secretions from the mycelia, as well as three combinations of fractions, were used in a cell cytotoxicity assay. Results from the assay suggest that secretions from H. erinaceus may be efficacious in causing U-87 MG cells to undergo apoptosis as compared to nontumorigenic MCF10A epithelial cells.
Pritika Shree Thotakura – DNA Barcoding as IdentificationBees play a crucial role in pollinating crops such as almonds and strawberries. However, the success of crop yields not only depends on the number of bees but also on the diversity of bee species in the area. Bee species differ in their behavior and plant preferences, which affects their pollination success. Therefore, it is essential to understand the variety of bees in an area to ensure agricultural success. To measure bee diversity, researchers can use a tool called DNA barcoding, which assigns a unique "barcode" to each species based on sequences with high interspecies variation but low intraspecies variation. This allows for highly accurate identification based on DNA sequences alone. By creating a database of these unique DNA sequences, researchers can identify the different species of bees in a specific area accurately. This can help reduce the number of misidentifications that may arise from morphological identification. Combining morphological identifications with barcoding identifications can also help reaffirm the efficacy of using morphology to identify bees. Our study aims to determine the DNA barcodes of bee species collected in Grayson Co., TX, and add them to the Barcode of Life Data Systems (BOLD) database. By contributing to these databases, a more thorough understanding of the local North Texas bee species may be achieved.
