Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.984
Lindsey Morais, Daron Weekley, Holly L. Racine
Craniosynostosis (CS) is the premature fusion of the cranial sutures during embryonic development, and this results in intracranial pressure and skull deformities. Thyrotoxicosis, specifically maternal hyperthyroidism, has been linked to the development of CS. Our lab is currently validating an avian model of thyroxine-induced CS to study the mechanism involved in thyroxine-enhanced cranial ossification. We hypothesized that thyroxine exposure will alter skull morphology. Two groups of fertilized chicken eggs were injected with saline or 25 ng T4 into the air cell on embryonic days (E) 11 and 15. A set of skulls from each group were collected on E17-19 (n = 17-21/day). They were fixed in formalin and processed using Alizarin Red whole-mount staining, imaged (superior view), and quantified to determine thyroxine-induced alteration in skull morphology. Geometric morphometric analysis using MorphoJ was performed to identify shape variation between treatment groups of the fibrous space between ossifying regions of the anterior fontanelle. These preliminary results demonstrated a significant variation in the shape of the fibrous gap between developing sutures on embryonic days 17, 18, and 19 (p<0.05). Using the same methodology, an additional set of skulls on E19 were collected from each group and processed using both Alizarin red and Alcian blue whole-mount staining to better delineate the ossifying region of bone. Analysis is currently in progress. In conclusion, these findings support that our model is successful in promoting the fusing of the cranial bones with thyroxine exposure in our avian model through altered skull morphology visualized in the anterior fontanelle.
颅缝闭合(CS)是胚胎发育过程中颅缝的过早融合，导致颅内压和颅骨畸形。甲状腺毒症，特别是母亲甲状腺功能亢进，与CS的发展有关。我们的实验室目前正在验证甲状腺素诱导CS的鸟类模型，以研究甲状腺素增强颅骨骨化的机制。我们假设甲状腺素暴露会改变颅骨形态。两组受精卵在胚胎第11和15天分别向空气细胞注射生理盐水或25 ng T4。各组颅骨于E17-19日各取一组(n = 17-21/天)。将它们固定在福尔马林中，用茜素红整片染色处理，成像(上位视图)，并量化以确定甲状腺素引起的颅骨形态学改变。使用MorphoJ进行几何形态计量学分析，以确定前囟门骨化区之间纤维间隙在治疗组之间的形状变化。这些初步结果表明，在胚胎17,18和19天，发育缝线之间的纤维间隙形状发生了显著变化(p<0.05)。使用相同的方法，从每组E19上收集另一组头骨，并使用茜素红和阿利新蓝全片染色进行处理，以更好地描绘骨骼的骨化区域。分析目前正在进行中。总之，这些发现支持我们的模型是成功地促进颅骨融合与甲状腺素暴露在我们的鸟类模型中，通过改变颅骨形态学在前囟门可见。
{"title":"Differential morphometric analysis of the anterior fontanelle in an embryonic avian model of induced-thyrotoxicosis.","authors":"Lindsey Morais, Daron Weekley, Holly L. Racine","doi":"10.55632/pwvas.v95i2.984","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.984","url":null,"abstract":"Craniosynostosis (CS) is the premature fusion of the cranial sutures during embryonic development, and this results in intracranial pressure and skull deformities. Thyrotoxicosis, specifically maternal hyperthyroidism, has been linked to the development of CS. Our lab is currently validating an avian model of thyroxine-induced CS to study the mechanism involved in thyroxine-enhanced cranial ossification. We hypothesized that thyroxine exposure will alter skull morphology. Two groups of fertilized chicken eggs were injected with saline or 25 ng T4 into the air cell on embryonic days (E) 11 and 15. A set of skulls from each group were collected on E17-19 (n = 17-21/day). They were fixed in formalin and processed using Alizarin Red whole-mount staining, imaged (superior view), and quantified to determine thyroxine-induced alteration in skull morphology. Geometric morphometric analysis using MorphoJ was performed to identify shape variation between treatment groups of the fibrous space between ossifying regions of the anterior fontanelle. These preliminary results demonstrated a significant variation in the shape of the fibrous gap between developing sutures on embryonic days 17, 18, and 19 (p<0.05). Using the same methodology, an additional set of skulls on E19 were collected from each group and processed using both Alizarin red and Alcian blue whole-mount staining to better delineate the ossifying region of bone. Analysis is currently in progress. In conclusion, these findings support that our model is successful in promoting the fusing of the cranial bones with thyroxine exposure in our avian model through altered skull morphology visualized in the anterior fontanelle.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87389831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.974
Annalee Corcoran, Jason Rafe Miller
Data preparation is a critical step for any machine learning experiment. We have analyzed a dataset derived from images of human male lung cancer tumors. These tumors had been analyzed with genetic markers to identify Y-chromosome loss, which was the case in about half of the samples. Whole slide images (WSI) had been collected and H&E stained by collaborators. We had processed the images with the CellProfiler software to extract numeric features. In this study, we analyzed the data in preparation for training a convolutional neural network to predict Y-chromosome loss from the extracted features, thereby recapitulating the genetic marker analysis. Using Excel and Python, we identified uninformative features and missing data. We predict that data cleaning, informed by these results, will improve the chances of successful machine learning.
{"title":"Analyzing Lung Cancer Data for Machine Learning","authors":"Annalee Corcoran, Jason Rafe Miller","doi":"10.55632/pwvas.v95i2.974","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.974","url":null,"abstract":"Data preparation is a critical step for any machine learning experiment. We have analyzed a dataset derived from images of human male lung cancer tumors. These tumors had been analyzed with genetic markers to identify Y-chromosome loss, which was the case in about half of the samples. Whole slide images (WSI) had been collected and H&E stained by collaborators. We had processed the images with the CellProfiler software to extract numeric features. In this study, we analyzed the data in preparation for training a convolutional neural network to predict Y-chromosome loss from the extracted features, thereby recapitulating the genetic marker analysis. Using Excel and Python, we identified uninformative features and missing data. We predict that data cleaning, informed by these results, will improve the chances of successful machine learning.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"2014 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73450415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.1012
Sue Newberry
I worked with the non-profit organization Eco Vrindavan Inc., located in Moundsville West Virginia to grow and provide organic food to the local community. Wheeling’s Grow Ohio Valley, an organization that supports local food production, assisted in installing “hoop houses” which increase the length of the growing season and increase production. Hoop houses are unheated structures covered in plastic where crops are planted directly into the ground. While working for Eco Vrindavan Inc., I observed how hoop houses extend the growing season by keeping the ground from freezing and ensuring the survival of selected crops through winter months. During this internship, I grew, planted, and harvested various greens in order to provide produce to the community. I learned what types of greens do well in cold climates and which ones do not and I have gained experience in how to organically fertilize plants, check soil quality, and look for mites and other diseases on crops. Currently, preparation for spring planting is underway to ensure early crops of flowers and summer vegetables. Hoop houses may provide benefits to organic growers who need crops to be available all year long and help minimize labor costs by keeping pests away while ensuring temperature changes do not negatively affect crops. In the future, I hope to educate other organic growers on the benefits hoop houses provide and use what I have learned to introduce organic growing in hoop houses to more communities statewide.
{"title":"Hoop Houses Provide Optimal Conditions for Increased Crop Production Year-round","authors":"Sue Newberry","doi":"10.55632/pwvas.v95i2.1012","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1012","url":null,"abstract":"I worked with the non-profit organization Eco Vrindavan Inc., located in Moundsville West Virginia to grow and provide organic food to the local community. Wheeling’s Grow Ohio Valley, an organization that supports local food production, assisted in installing “hoop houses” which increase the length of the growing season and increase production. Hoop houses are unheated structures covered in plastic where crops are planted directly into the ground. While working for Eco Vrindavan Inc., I observed how hoop houses extend the growing season by keeping the ground from freezing and ensuring the survival of selected crops through winter months. During this internship, I grew, planted, and harvested various greens in order to provide produce to the community. I learned what types of greens do well in cold climates and which ones do not and I have gained experience in how to organically fertilize plants, check soil quality, and look for mites and other diseases on crops. Currently, preparation for spring planting is underway to ensure early crops of flowers and summer vegetables. Hoop houses may provide benefits to organic growers who need crops to be available all year long and help minimize labor costs by keeping pests away while ensuring temperature changes do not negatively affect crops. In the future, I hope to educate other organic growers on the benefits hoop houses provide and use what I have learned to introduce organic growing in hoop houses to more communities statewide.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"61 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84244503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.953
R. Harsh, L. Robertson
Fungi are ubiquitous and their spores are found in the air outside and within the built environment. We investigated the fungal species present in a teaching laboratory at Shepherd University. Airborne fungi were captured passively at three sites within the teaching laboratory on two different culture media. Viable fungi were isolated to pure culture and then identified by DNA barcode using the internal transcribed spacer region of the ribosomal DNA. Twenty-four isolates were identified to order, family, genus, or species; most isolates were not resolved to species by DNA barcode. Captured fungi belonged to Helotiales (1 isolate), Pleosporales (2 isolates), Hypocreales (1 isolate), Didymellaceae (1 isolate), Cladosporiaceae (5 isolates), Aspergillaceae (1 isolate), Sporocadaceae (1 isolate), Peniophora (1 isolate), Trichoderma (1 isolate), Penicillium (4 isolates), Fusarium (2 isolates), and Myrmecridium (1 isolate). Three fungal isolates were identified as Xenoacrodontium juglandis. These results are similar to other studies investigating the fungal community in the built environment, where Cladosporium, Penicillium, and Aspergillus are the most commonly identified genera.
{"title":"Identification by DNA barcode of viable fungi captured in a teaching laboratory in West Virginia","authors":"R. Harsh, L. Robertson","doi":"10.55632/pwvas.v95i2.953","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.953","url":null,"abstract":"Fungi are ubiquitous and their spores are found in the air outside and within the built environment. We investigated the fungal species present in a teaching laboratory at Shepherd University. Airborne fungi were captured passively at three sites within the teaching laboratory on two different culture media. Viable fungi were isolated to pure culture and then identified by DNA barcode using the internal transcribed spacer region of the ribosomal DNA. Twenty-four isolates were identified to order, family, genus, or species; most isolates were not resolved to species by DNA barcode. Captured fungi belonged to Helotiales (1 isolate), Pleosporales (2 isolates), Hypocreales (1 isolate), Didymellaceae (1 isolate), Cladosporiaceae (5 isolates), Aspergillaceae (1 isolate), Sporocadaceae (1 isolate), Peniophora (1 isolate), Trichoderma (1 isolate), Penicillium (4 isolates), Fusarium (2 isolates), and Myrmecridium (1 isolate). Three fungal isolates were identified as Xenoacrodontium juglandis. These results are similar to other studies investigating the fungal community in the built environment, where Cladosporium, Penicillium, and Aspergillus are the most commonly identified genera.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"16 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88457656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.998
Caleb Fritz
Pseudomonas aeruginosa is an opportunistic pathogen that affects many patients in a hospital setting as well as immunocompromised patients. Pseudomonas aeruginosa is resistant to many antibiotics because of the formation of biofilms and pigmentations serving as virulence factors. Pseudomonas aeruginosa isolates are either mucoid or alginate-producing and non-alginate-producing (non-mucoid) isolates. The main objective of this experiment was to analyze the pathogenicity of mucoid and non-mucoid isolates in P. aeruginosa in a mouse model. We hypothesized that mucoid isolates are more pathogenic compared to non-pathogenic ones. Pseudomonas aeruginosa isolates PAO1 (non-mucoid) and 2192 (mucoid) were used intranasally to infect and monitor mortality rates. Isolates starved in water for at least three months were tested for infectivity. The mucoid isolate 2192 showed a 67% death rate compared to 33% of PAO1, suggesting the mucoid is more pathogenic to BALB/c mice than the non-mucoid PAO1. The death rate of mice infected with strain DBA susceptible to P. aeruginosa infection was higher than that of the BALB/c strain. The results also suggest that non-starved bacteria are more pathogenic to mice than starved bacteria. We extend our study by infecting mouse strains under hindlimb-unloading (HU) conditions with P. aeruginosa mucoid and non-mucoid isolates. Our results show that HU conditions enhance infection, suggesting the effect of stress and physiological changes could lead to susceptibility to infection. Our data indicate that the mucoid isolate is more likely to kill all mouse strains.
{"title":"Non-starved mucoid Pseudomonas aeruginosa isolates cause a high death rate in strains of mice.","authors":"Caleb Fritz","doi":"10.55632/pwvas.v95i2.998","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.998","url":null,"abstract":"Pseudomonas aeruginosa is an opportunistic pathogen that affects many patients in a hospital setting as well as immunocompromised patients. Pseudomonas aeruginosa is resistant to many antibiotics because of the formation of biofilms and pigmentations serving as virulence factors. Pseudomonas aeruginosa isolates are either mucoid or alginate-producing and non-alginate-producing (non-mucoid) isolates. The main objective of this experiment was to analyze the pathogenicity of mucoid and non-mucoid isolates in P. aeruginosa in a mouse model. We hypothesized that mucoid isolates are more pathogenic compared to non-pathogenic ones. Pseudomonas aeruginosa isolates PAO1 (non-mucoid) and 2192 (mucoid) were used intranasally to infect and monitor mortality rates. Isolates starved in water for at least three months were tested for infectivity. The mucoid isolate 2192 showed a 67% death rate compared to 33% of PAO1, suggesting the mucoid is more pathogenic to BALB/c mice than the non-mucoid PAO1. The death rate of mice infected with strain DBA susceptible to P. aeruginosa infection was higher than that of the BALB/c strain. The results also suggest that non-starved bacteria are more pathogenic to mice than starved bacteria. We extend our study by infecting mouse strains under hindlimb-unloading (HU) conditions with P. aeruginosa mucoid and non-mucoid isolates. Our results show that HU conditions enhance infection, suggesting the effect of stress and physiological changes could lead to susceptibility to infection. Our data indicate that the mucoid isolate is more likely to kill all mouse strains. ","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"46 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86359332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.978
Reagan Gray
Methicillin-Resistant Staphylococcus aureus (MRSA) is an antibiotic-resistant bacterium and a major cause of preventable illness and death. To combat MRSA and other antibiotic-resistant pathogens, alternatives to small molecule therapeutics targeting essential biological pathways of bacteria must be identified. Staphylococci, including MRSA strains, can exist as part of the human microbiota, competing for resources and space with other microorganisms. Here, we sought to identify bacteria from the human microbiota capable of killing or inhibiting the growth of MRSA. Swabs of the nostrils and skin of two individuals were collected, one being a natural MRSA carrier, and bacteria were isolated from these samples. Of these isolates, eight showed to inhibit the growth of a MRSA strain when cross streaking. To identify these bacteria, a portion of the 16s rRNA gene of these bacteria was sequenced and compared to known bacteria in the genbank database using BLAST. Of the eight isolates, three distinct bacterial species were identified: Acinetobacter baumannii, Bacillus aerius, and Staphylococcus epidermidis. Following the identification of these bacteria, we sought to determine the mechanism of inhibition. Cell lysates and extracellular material of these inhibitory bacteria were used in a disk diffusion assay which showed no observable zones of inhibition toward MRSA. This suggests that these bacteria may need to be viable and/or directly contact MRSA to mediate the inhibition of growth. To test this hypothesis, phase-contrast microscopy of co-cultured bacteria was used. While results here suggested that B. aerius directly interfered with the ability of MRSA to replicate, future studies using time-lapse microscopy of fluorescent strains of these microbes should produce data that can be more clearly interpreted.
{"title":"The identification of bacteria from human microbiota that inhibit the growth of Methicillin-Resistant Staphylococcus aureus","authors":"Reagan Gray","doi":"10.55632/pwvas.v95i2.978","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.978","url":null,"abstract":"Methicillin-Resistant Staphylococcus aureus (MRSA) is an antibiotic-resistant bacterium and a major cause of preventable illness and death. To combat MRSA and other antibiotic-resistant pathogens, alternatives to small molecule therapeutics targeting essential biological pathways of bacteria must be identified. Staphylococci, including MRSA strains, can exist as part of the human microbiota, competing for resources and space with other microorganisms. Here, we sought to identify bacteria from the human microbiota capable of killing or inhibiting the growth of MRSA. Swabs of the nostrils and skin of two individuals were collected, one being a natural MRSA carrier, and bacteria were isolated from these samples. Of these isolates, eight showed to inhibit the growth of a MRSA strain when cross streaking. To identify these bacteria, a portion of the 16s rRNA gene of these bacteria was sequenced and compared to known bacteria in the genbank database using BLAST. Of the eight isolates, three distinct bacterial species were identified: Acinetobacter baumannii, Bacillus aerius, and Staphylococcus epidermidis. Following the identification of these bacteria, we sought to determine the mechanism of inhibition. Cell lysates and extracellular material of these inhibitory bacteria were used in a disk diffusion assay which showed no observable zones of inhibition toward MRSA. This suggests that these bacteria may need to be viable and/or directly contact MRSA to mediate the inhibition of growth. To test this hypothesis, phase-contrast microscopy of co-cultured bacteria was used. While results here suggested that B. aerius directly interfered with the ability of MRSA to replicate, future studies using time-lapse microscopy of fluorescent strains of these microbes should produce data that can be more clearly interpreted. ","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90285420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.997
Emily Shupe
Chronic stress is known to suppress the immune system, but the influence of stress on the pathogenesis of Chlamydia trachomatis genital infection and host immune response remains unexplored A stress mouse model has been developed in our lab and we have demonstrated that cold-induced stress (CIS) suppresses the immune system and subsequently leads to increased intensity of Chlamydia muridaum in mice. We have started feeding mice with Active hexose correlated compound (AHCC) that may restore the function of the immune system and result in reduced C. muridarum shedding from the genital tract. During the feeding of stressed mice with AHCC, dendritic cells (DCs) and macrophages bring the AHCC to T cells in the lymph node; however, the specific functions of DCs subsets in our stress model are not defined. We determined the production of cytokines by macrophages, CDs, and CD4+T in the AHCC-fed stress mice during C. muridarum genital infection. CD4+ T cells isolated from the spleen or lymph node of stressed-AHCC-fed, PBS-stressed, or non-stressed mice proliferated, and cytokine secretion was measured by ELISA. AHCC-feeding led to lower production of IL-5, IL-13, IL-10 and higher IL-12 (P<0.01) and IFN-
{"title":"Active Hexose Correlated Compound as a Countermeasure against Cold-induced Stress during Chlamydia muridarum Genital Infection in Mice.","authors":"Emily Shupe","doi":"10.55632/pwvas.v95i2.997","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.997","url":null,"abstract":"Chronic stress is known to suppress the immune system, but the influence of stress on the pathogenesis of Chlamydia trachomatis genital infection and host immune response remains unexplored A stress mouse model has been developed in our lab and we have demonstrated that cold-induced stress (CIS) suppresses the immune system and subsequently leads to increased intensity of Chlamydia muridaum in mice. We have started feeding mice with Active hexose correlated compound (AHCC) that may restore the function of the immune system and result in reduced C. muridarum shedding from the genital tract. During the feeding of stressed mice with AHCC, dendritic cells (DCs) and macrophages bring the AHCC to T cells in the lymph node; however, the specific functions of DCs subsets in our stress model are not defined. We determined the production of cytokines by macrophages, CDs, and CD4+T in the AHCC-fed stress mice during C. muridarum genital infection. CD4+ T cells isolated from the spleen or lymph node of stressed-AHCC-fed, PBS-stressed, or non-stressed mice proliferated, and cytokine secretion was measured by ELISA. AHCC-feeding led to lower production of IL-5, IL-13, IL-10 and higher IL-12 (P<0.01) and IFN-","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84938919","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.1013
Sara Roberson, Ryan M. Anderson, Catherine Stress
The habenula is a conserved region of the brain that affects emotion and behavior, and is located in the dorsal diencephalon of zebrafish. Previous work has shown that chemokine signaling plays a significant role in the direction of habenular axon growth. The chemokine receptor cxcr4b is expressed in habenular precursors and the Cxcr4b protein is found on the newly forming habenular axons. Chemokine ligands Cxcl12a and Cxcl12b are both found in the region around the developing habenula. The focus of this research is to examine the relationship of these two chemokines to determine their respective roles in directing axon outgrowth. The relative expression domains of the two chemokine ligands are of interest, as it is unknown if their expression domains overlap. Additionally, we are identifying fish that are heterozygous for a mutation in the cxcl12b gene to examine habenular axons in larvae lacking functional Cxcl12b. To explore the role of Cxc12a we have performed overexpression experiments using a heat shock protocol in fish carrying a fluorescent marker of the habenular and its axons. However, the current transgenic method to confirm overexpression of cxcl12a interferes with visualization of the habenular region. We plan to employ a chemical protocol to validate disruption of chemokine signaling that does not compromise observation of the habenula and its axons.
{"title":"The role of chemokine signaling in habenular axon projection in zebrafish","authors":"Sara Roberson, Ryan M. Anderson, Catherine Stress","doi":"10.55632/pwvas.v95i2.1013","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1013","url":null,"abstract":"The habenula is a conserved region of the brain that affects emotion and behavior, and is located in the dorsal diencephalon of zebrafish. Previous work has shown that chemokine signaling plays a significant role in the direction of habenular axon growth. The chemokine receptor cxcr4b is expressed in habenular precursors and the Cxcr4b protein is found on the newly forming habenular axons. Chemokine ligands Cxcl12a and Cxcl12b are both found in the region around the developing habenula. The focus of this research is to examine the relationship of these two chemokines to determine their respective roles in directing axon outgrowth. The relative expression domains of the two chemokine ligands are of interest, as it is unknown if their expression domains overlap. Additionally, we are identifying fish that are heterozygous for a mutation in the cxcl12b gene to examine habenular axons in larvae lacking functional Cxcl12b. To explore the role of Cxc12a we have performed overexpression experiments using a heat shock protocol in fish carrying a fluorescent marker of the habenular and its axons. However, the current transgenic method to confirm overexpression of cxcl12a interferes with visualization of the habenular region. We plan to employ a chemical protocol to validate disruption of chemokine signaling that does not compromise observation of the habenula and its axons.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"47 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76206522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.971
Hannah Bonham
This project studies the effects of time and hand dominance on different characteristics of handwriting analysis. The handwriting characteristics that were tested are line quality, slope of writing, pen lifts, letter spacing, and word spacing. These characteristics were tested in a subject’s dominant and non-dominant handwriting. Twenty-five adult human subjects participated in this experiment. The research was split into two parts. Part one analyzed the characteristics of a subject’s dominant and non-handwriting during the morning, afternoon, and evening of (preferably) the same day. In the part one phase, participants were asked to write: "The quick brown fox jumps over the lazy dog." Part two analyzed the authenticity of a freehand, simulation, and tracing forgery. A ruler was used to quantify the handwriting characteristics. The line quality of the dominant and non-dominant hands does not change during the different sessions. The slope of the writing changes more with the non-dominant hand. The non-dominant hand has greater letter and word spacing. The dominant hand typically changed the pen lifts number more than the non-dominant hand. “Quick” was the word that had the most changes in pen lifts for both the dominant and non-dominant hand. The freehand forgery is easy to classify as a forged attempt. The simulation forgery can be distinguished from the genuine signature, but it can still be identified as a forgery. The traced forgery could pass off as a genuine signature.
{"title":"External effects on the characteristics of handwriting analysis and the analysis of forgery types.","authors":"Hannah Bonham","doi":"10.55632/pwvas.v95i2.971","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.971","url":null,"abstract":"This project studies the effects of time and hand dominance on different characteristics of handwriting analysis. The handwriting characteristics that were tested are line quality, slope of writing, pen lifts, letter spacing, and word spacing. These characteristics were tested in a subject’s dominant and non-dominant handwriting. Twenty-five adult human subjects participated in this experiment. The research was split into two parts. Part one analyzed the characteristics of a subject’s dominant and non-handwriting during the morning, afternoon, and evening of (preferably) the same day. In the part one phase, participants were asked to write: \"The quick brown fox jumps over the lazy dog.\" Part two analyzed the authenticity of a freehand, simulation, and tracing forgery. A ruler was used to quantify the handwriting characteristics. The line quality of the dominant and non-dominant hands does not change during the different sessions. The slope of the writing changes more with the non-dominant hand. The non-dominant hand has greater letter and word spacing. The dominant hand typically changed the pen lifts number more than the non-dominant hand. “Quick” was the word that had the most changes in pen lifts for both the dominant and non-dominant hand. The freehand forgery is easy to classify as a forged attempt. The simulation forgery can be distinguished from the genuine signature, but it can still be identified as a forgery. The traced forgery could pass off as a genuine signature.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"52 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79037160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.964
Elio Delatore III, Joseph Horzempa, Stuart Cantlay, Elle Roberts
Francisella tularensis is a bacterium that induces the zoonotic disease tularemia. In the course of infection, F. tularensis bacteria invade erythrocytes, a phenomenon that heightens the colonization of ticks after a blood meal. To better understand the mechanism of erythrocyte invasion, we hypothesized that transcription of bacterial genes significant in erythrocyte invasion would be upregulated upon exposure to these host cells. An RNA-seq unveiled that transcription of 7% of F. tularensis genes augment when in erythrocyte presence. Of these, we pinpointed a putative transcriptional regulator, FTL_1199. The goal was to delete FTL_1199 in F. tularensis LVS. SOE PCR amplified and duplicated the up and downstream regions of the target gene in tandem into a shuttle vector that is insecure within F. tularensis. This newly generated plasmid, pDEL1199, was mobilized inside of F. tularensis by conjugation. Merodiploid strains generated by homologous recombination were isolated and transformed with pGUTS. Expression of I-Sce1 within the merodiploid produces a double-stranded break. This breakage resulted in a second recombination that either ensued to wild-type or deletion of FTL_1199 deduced through a PCR. Finally, in DFTL_1199 strains, pGUTS was cured by successive cultivation in the absence of selection followed by replica-plating on chocolate II agar ± kanamycin. Gentamicin protection assays showed reduced levels of erythrocyte invasion for F. tularensis DFTL_1199 compared to wild type bacteria. However, complementation of FTL_1199 to the deletion mutant restored this strain’s ability to invade red blood cells. These findings demonstrate that FTL_1199 is important for erythrocyte invasion by F. tularensis.
(Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence, R15HL14735 from NHLBI, and funds from the NASA West Virginia Space Grant Consortium).
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 (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence, R15HL14735 from NHLBI, and funds from the NASA West Virginia Space Grant Consortium).","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135972603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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