Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.1011
M. Hall
Antibiotic resistance is an urgent public health threat. The CDC estimates there are approximately 2.8 million new cases of antibiotic-resistant infections annually resulting in 35,000 deaths and billions of dollars in health care costs. The development of new drugs is imperative to combat this crisis and prevent the loss of additional lives from once “curable” diseases. Resazomycins, a novel family of antibiotics, have bactericidal activity against Francisella tularensis and Neisseria gonorrhoeae. One resazomycin, resorufin pentyl ether (RPE), significantly reduces vaginal colonization by N. gonorrhoeae in a mouse model of infection. Repeated administration of RPE, however, fails to clear the infection, in contrast to a single dose of ceftriaxone, an antibiotic commonly used to treat gonorrhea, which clears the infection within 24 hours. Further characterization of resazomycins revealed the efficacy of these compounds is limited by interaction with serum albumin and reduced oxygen concentrations found within mammalian tissues. Therefore, we hypothesize that novel resazurin analogs that maintain antimicrobial activity in the presence of serum albumin and low oxygen will have improved therapeutic efficacy in vivo. To date, two different derivatives of RPE have been synthesized and tested for antimicrobial activity against F. tularensis and N. gonorrhoeae – 1-methyl RPE and 4-methyl RPE. Neither of these compounds inhibited the growth of F. tularensis or N. gonorrhoeae. Next, we plan to prepare a series of ketone derivatives of resazurin to alter the electrophilicity and reduction potential of these compounds and test their efficacy against F. tularensis and N. gonorrhoeae.
{"title":"Screening Resorufin Pentyl Ether Analogs for Enhanced Antimicrobial Activity Against Francisella tularensis and Neisseria gonorrhoeae","authors":"M. Hall","doi":"10.55632/pwvas.v95i2.1011","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1011","url":null,"abstract":"Antibiotic resistance is an urgent public health threat. The CDC estimates there are approximately 2.8 million new cases of antibiotic-resistant infections annually resulting in 35,000 deaths and billions of dollars in health care costs. The development of new drugs is imperative to combat this crisis and prevent the loss of additional lives from once “curable” diseases. Resazomycins, a novel family of antibiotics, have bactericidal activity against Francisella tularensis and Neisseria gonorrhoeae. One resazomycin, resorufin pentyl ether (RPE), significantly reduces vaginal colonization by N. gonorrhoeae in a mouse model of infection. Repeated administration of RPE, however, fails to clear the infection, in contrast to a single dose of ceftriaxone, an antibiotic commonly used to treat gonorrhea, which clears the infection within 24 hours. Further characterization of resazomycins revealed the efficacy of these compounds is limited by interaction with serum albumin and reduced oxygen concentrations found within mammalian tissues. Therefore, we hypothesize that novel resazurin analogs that maintain antimicrobial activity in the presence of serum albumin and low oxygen will have improved therapeutic efficacy in vivo. To date, two different derivatives of RPE have been synthesized and tested for antimicrobial activity against F. tularensis and N. gonorrhoeae – 1-methyl RPE and 4-methyl RPE. Neither of these compounds inhibited the growth of F. tularensis or N. gonorrhoeae. Next, we plan to prepare a series of ketone derivatives of resazurin to alter the electrophilicity and reduction potential of these compounds and test their efficacy against F. tularensis and N. gonorrhoeae.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"119 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90264947","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.v95i2.963
Sana Mazhar
Drug Discovery Research (DDR) is a process by which new potential therapeutic entities are recognized. Lead discovery and Computer aided drug discovery are two most popular methods in DDR. However, chemical modification such as development of the chemical linker gains its popularity in various therapeutic classes. In this poster, we are proposing four chemical steps starting from commercially available amino acids and their chemical routes for synthesizing linkers. We hypothesize that the more sterically hindered R group on Amino Acid will result in an increase in diastereomer ratio (DR) in reduction chemistry. In this poster we will discuss the detailed chemical reaction methods, their data, and how it’s being proposed as a potential anti-cancer agent.
{"title":"Study of Steric Hindrance of Ketone Ester Reduction Chemistry","authors":"Sana Mazhar","doi":"10.55632/pwvas.v95i2.963","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.963","url":null,"abstract":"Drug Discovery Research (DDR) is a process by which new potential therapeutic entities are recognized. Lead discovery and Computer aided drug discovery are two most popular methods in DDR. However, chemical modification such as development of the chemical linker gains its popularity in various therapeutic classes. In this poster, we are proposing four chemical steps starting from commercially available amino acids and their chemical routes for synthesizing linkers. We hypothesize that the more sterically hindered R group on Amino Acid will result in an increase in diastereomer ratio (DR) in reduction chemistry. In this poster we will discuss the detailed chemical reaction methods, their data, and how it’s being proposed as a potential anti-cancer agent. ","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"44 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88282067","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.v95i2.1010
Kimberly, Kristy
This research is useful in forensic laboratories to prevent contamination. If a cleaner was found to remove all traces of blood, it could be used to sterilize lab benches and equipment to ensure no cross contamination between pieces of evidence. It could also be used in the field to determine which presumptive test would be more useful in different scenarios. The purpose of this project was to determine if common household cleaners could efficiently remove the presence of blood. To test this theory, I used hydrogen peroxide cleaner, dawn dish soap, and Clorox bleach to clean blood from three surfaces, laminate flooring, textured glass, and carpet. This was done using two different cleaning materials, paper towels and microfiber cloths, at 3-time intervals. After removing the visual evidence of blood, I tested for residual blood using two different presumptive tests, tetramethylbenzidine and phenolphthalein. Results show that three out of the twelve variable combinations gave all positive results. Over 50% of the presumptive tests ran on textured glass were negative. From the results of the presumptive tests, it was found that the tetramethylbenzidine test was more sensitive than the phenolphthalein test. Household cleaners do not effectively clean up blood, leaving behind enough residual blood to test positive using presumptive blood tests. The tetramethylbenzidine test was more sensitive than the other. More variables should be tested to determine the effects of household cleaners on surfaces such as furniture. Different presumptive tests should also be examined.
{"title":"Effects of Household Cleaning Supplies on Trace Blood Evidence","authors":"Kimberly, Kristy","doi":"10.55632/pwvas.v95i2.1010","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1010","url":null,"abstract":"This research is useful in forensic laboratories to prevent contamination. If a cleaner was found to remove all traces of blood, it could be used to sterilize lab benches and equipment to ensure no cross contamination between pieces of evidence. It could also be used in the field to determine which presumptive test would be more useful in different scenarios. The purpose of this project was to determine if common household cleaners could efficiently remove the presence of blood. To test this theory, I used hydrogen peroxide cleaner, dawn dish soap, and Clorox bleach to clean blood from three surfaces, laminate flooring, textured glass, and carpet. This was done using two different cleaning materials, paper towels and microfiber cloths, at 3-time intervals. After removing the visual evidence of blood, I tested for residual blood using two different presumptive tests, tetramethylbenzidine and phenolphthalein. \u0000 Results show that three out of the twelve variable combinations gave all positive results. Over 50% of the presumptive tests ran on textured glass were negative. From the results of the presumptive tests, it was found that the tetramethylbenzidine test was more sensitive than the phenolphthalein test. Household cleaners do not effectively clean up blood, leaving behind enough residual blood to test positive using presumptive blood tests. The tetramethylbenzidine test was more sensitive than the other. More variables should be tested to determine the effects of household cleaners on surfaces such as furniture. Different presumptive tests should also be examined.","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":"89088804","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.v95i2.1016
Elizabeth Walters
Growing concern about climate change created the need for a globally applied system of accounting to track demand on the earth’s resources and their supply. An ecological footprint analysis provides an objective metric for the ecological impact a nation has on the environment, measured in global hectares/capita. Using data provided by the Global footprint Network, we calculate that the UAE has an ecological deficit of 9.2 global hectares/person but has only 9% of the area required to offset the cost of its environmental impact. The UAE is working to reduce this deficit and can approach this by 1) increasing its biocapacity and 2) reducing its ecological footprint. A Pearson Correlation Coefficient was computed to assess the linear relationship between each of the 6 land use metrics described by Wackernagel (1990) and the total ecological footprint of the UAE. Loss of fisheries was found to significantly impact the biocapacity, r(37) = [.998], p= [1.3347E-53], while carbon emissions have contributed the most to the ecological footprint r(37) = [ .986], p = [ 6.7143E-36]. These data can inform options for the targeted support one of earth’s most vulnerable ecosystems and encourage increased use of renewable energy sources. This project is supported by the U.S. Dept. of State Diplomacy Lab (#2209921) and Bluefield State University International Initiatives.
{"title":"Assessing the UAE's Efforts to Combat Climate Change using Ecological Footprint Accounting","authors":"Elizabeth Walters","doi":"10.55632/pwvas.v95i2.1016","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1016","url":null,"abstract":"\u0000 \u0000 \u0000Growing concern about climate change created the need for a globally applied system of accounting to track demand on the earth’s resources and their supply. An ecological footprint analysis provides an objective metric for the ecological impact a nation has on the environment, measured in global hectares/capita. Using data provided by the Global footprint Network, we calculate that the UAE has an ecological deficit of 9.2 global hectares/person but has only 9% of the area required to offset the cost of its environmental impact. The UAE is working to reduce this deficit and can approach this by 1) increasing its biocapacity and 2) reducing its ecological footprint. A Pearson Correlation Coefficient was computed to assess the linear relationship between each of the 6 land use metrics described by Wackernagel (1990) and the total ecological footprint of the UAE. Loss of fisheries was found to significantly impact the biocapacity, r(37) = [.998], p= [1.3347E-53], while carbon emissions have contributed the most to the ecological footprint r(37) = [ .986], p = [ 6.7143E-36]. These data can inform options for the targeted support one of earth’s most vulnerable ecosystems and encourage increased use of renewable energy sources. This project is supported by the U.S. Dept. of State Diplomacy Lab (#2209921) and Bluefield State University International Initiatives. \u0000 \u0000 \u0000","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"42 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82331183","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.v95i2.1009
Caitlynn Morningstar
Resistance to antibiotic treatments coupled with the decline in antibiotic discovery has resulted in a steady increase in deaths caused by once “curable” bacterial infections. Developing new drugs is crucial to prevent more loss of life in the future. We discovered the compound resazurin exhibits antimicrobial activity against gram-negative bacteria including Francisella tularensis (Ft), however, certain strains of Ft have developed resistance to resazurin. Understanding how Ftdevelops resistance to resazurin will help with defining the mechanism by which resazurin elicits its antimicrobial effect. Whole genome sequencing of resazurin-resistant (Rzr) Ft LVS mutants revealed 93% of the isolates sequenced possessed mutations in the coding regions of FTL_0421, FTL_0895, and FTL_1504. The focus of my project was to explore the role of FTL_0895 in resazurin susceptibility. To confirm this gene plays a role in the reduced susceptibility of the Rzr strains to resazurin, a wild-type copy of FTL_0895 was cloned into the Francisella shuttle vector pABST and thenelectroporated into one of the Rzr mutants, Rzr1. Complementation with FTL_0895 did not restore sensitivity of the Rzr1 strain to resazurin suggesting that mutation of this gene alone is not responsible for resistance to resazurin. Therefore, we have cloned FTL_0895 into a different Ft shuttle vector pMQ2 so we can complement back Rzr1 with this gene in combination with either FTL_1504 or FTL_0421. Agar dilution and time kill assays will be conducted on the resulting Rzr-complemented strains to determine their susceptibility to resazurin.
对抗生素治疗的耐药性，加上抗生素发现的减少，导致一度“可治愈”的细菌感染造成的死亡人数稳步增加。开发新药对于防止未来更多的生命损失至关重要。我们发现化合物reazurin对革兰氏阴性菌包括土拉菌Francisella tularensis (Ft)具有抗菌活性，然而，某些菌株对reazurin产生了耐药性。了解ftd如何产生对瑞祖林的耐药性将有助于确定瑞祖林引发其抗菌作用的机制。reazurin -resistant (Rzr) Ft LVS突变体的全基因组测序结果显示，93%的突变株在FTL_0421、FTL_0895和FTL_1504编码区存在突变。我的项目重点是探索FTL_0895在resazurin易感性中的作用。为了证实该基因在降低Rzr菌株对resazurin的易感性中起作用，将FTL_0895的野生型拷贝克隆到Francisella穿梭载体pABST中，然后电孔插入Rzr突变体Rzr1中。与FTL_0895的互补不能恢复Rzr1菌株对resazurin的敏感性，这表明该基因的突变不能单独对resazurin产生抗性。因此，我们将FTL_0895克隆到不同的Ft穿梭载体pMQ2中，这样我们就可以将该基因与FTL_1504或FTL_0421结合补充回Rzr1。琼脂稀释和时间杀伤试验将对得到的rzr补充菌株进行，以确定它们对reazurin的敏感性。
{"title":"Role of FTL_0895 in Francisella tularensis Susceptibility to Resazurin","authors":"Caitlynn Morningstar","doi":"10.55632/pwvas.v95i2.1009","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1009","url":null,"abstract":"Resistance to antibiotic treatments coupled with the decline in antibiotic discovery has resulted in a steady increase in deaths caused by once “curable” bacterial infections. Developing new drugs is crucial to prevent more loss of life in the future. We discovered the compound resazurin exhibits antimicrobial activity against gram-negative bacteria including Francisella tularensis (Ft), however, certain strains of Ft have developed resistance to resazurin. Understanding how Ftdevelops resistance to resazurin will help with defining the mechanism by which resazurin elicits its antimicrobial effect. Whole genome sequencing of resazurin-resistant (Rzr) Ft LVS mutants revealed 93% of the isolates sequenced possessed mutations in the coding regions of FTL_0421, FTL_0895, and FTL_1504. The focus of my project was to explore the role of FTL_0895 in resazurin susceptibility. To confirm this gene plays a role in the reduced susceptibility of the Rzr strains to resazurin, a wild-type copy of FTL_0895 was cloned into the Francisella shuttle vector pABST and thenelectroporated into one of the Rzr mutants, Rzr1. Complementation with FTL_0895 did not restore sensitivity of the Rzr1 strain to resazurin suggesting that mutation of this gene alone is not responsible for resistance to resazurin. Therefore, we have cloned FTL_0895 into a different Ft shuttle vector pMQ2 so we can complement back Rzr1 with this gene in combination with either FTL_1504 or FTL_0421. Agar dilution and time kill assays will be conducted on the resulting Rzr-complemented strains to determine their susceptibility to resazurin. ","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83205687","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.v95i2.968
Emma Beatty, Deanna M Schmitt
Francisella tularensis is defined by the Centers for Disease Control and Prevention as a Category A bioterrorism agent due to its low infectious dose, high mortality rate, and ease of aerosolization. Given there is no licensed tularemia vaccine in the United States and the possible development and release of antibiotic-resistant F. tularensis strains, there is an urgent need for new treatments against this bacterium. We determined the phenoxazine dye resazurin (Rz) exhibits antimicrobial activity against F. tularensis and other gram-negative bacteria. The mode of action of this compound is not understood, but potential targets of Rz were identified in a high throughput screen for resistant isolates. Ninety-three percent of the Rz-resistant (Rzr) isolates sequenced contained mutations within the coding regions of FTL_0421 (lpnA), FTL_0895, and FTL_1504 (katG). To confirm mutation of lpnA was contributing to Rz resistance, we introduced a wild-type copy of lpnA into Rzr1 and then assessed the susceptibility of the resulting complemented strain (Rzr1/pABST-lpnA). The minimum inhibitory concentration (MIC) of Rzr1 and Rzr1/pABST-lpnA were similar, therefore, we wanted to confirm we were restoring expression of LpnA in the complemented strain via Western blot analysis. As expected, LpnA was expressed in wild-type LVS but not in Rzr1 due to the genetic mutation. In the Rzr1 strain transformed with pABST-lpnA, we still did not observe expression of LpnA suggesting our complementation strategy was ineffective. In the future, we plan to take an alternative approach to investigate the role of LpnA in resazurin resistance by creating a lpnA deletion mutant.
{"title":"Role of LpnA in Francisella tularensis Susceptibility to Resazurin.","authors":"Emma Beatty, Deanna M Schmitt","doi":"10.55632/pwvas.v95i2.968","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.968","url":null,"abstract":"Francisella tularensis is defined by the Centers for Disease Control and Prevention as a Category A bioterrorism agent due to its low infectious dose, high mortality rate, and ease of aerosolization. Given there is no licensed tularemia vaccine in the United States and the possible development and release of antibiotic-resistant F. tularensis strains, there is an urgent need for new treatments against this bacterium. We determined the phenoxazine dye resazurin (Rz) exhibits antimicrobial activity against F. tularensis and other gram-negative bacteria. The mode of action of this compound is not understood, but potential targets of Rz were identified in a high throughput screen for resistant isolates. Ninety-three percent of the Rz-resistant (Rzr) isolates sequenced contained mutations within the coding regions of FTL_0421 (lpnA), FTL_0895, and FTL_1504 (katG). To confirm mutation of lpnA was contributing to Rz resistance, we introduced a wild-type copy of lpnA into Rzr1 and then assessed the susceptibility of the resulting complemented strain (Rzr1/pABST-lpnA). The minimum inhibitory concentration (MIC) of Rzr1 and Rzr1/pABST-lpnA were similar, therefore, we wanted to confirm we were restoring expression of LpnA in the complemented strain via Western blot analysis. As expected, LpnA was expressed in wild-type LVS but not in Rzr1 due to the genetic mutation. In the Rzr1 strain transformed with pABST-lpnA, we still did not observe expression of LpnA suggesting our complementation strategy was ineffective. In the future, we plan to take an alternative approach to investigate the role of LpnA in resazurin resistance by creating a lpnA deletion mutant.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"21 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75499758","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.v95i2.1004
Nathasha Woart
Chlamydia lung infection caused by Chlamydia trachomatis is a serious lung infection particularly in infants but there are a few studies about lung infection. Chlamydial genital and lung infections in mice is frequently done with Chlamydia muridarum. The purpose of our study was to explore the effect of stress in lung chlamydia infection in male mice. We hypothesize that stress changes the level of cytokine production in monocytes during lung infection. As a The result from these studies include counting of live cells, differentiation of proliferation of bone marrow derived monocytes where the result showed that non-stressed dendritic cells (DCs) WT mice and stressed macrophages (MO) WT mice had sufficient live cells to study. No difference in IL-b production of macrophages of stressed and non-stress mice was observed. Beta2-adrenergic receptor (b2-AR agonist (fenoterol) and antagonist (ICI 118 551) treatment resulted in difference in cytokine production the effect of agonists and antagonists. TNF-alpha production was high in stressed mice compared to non-stress mice for MO. LPS stimulated TNF-alpha production in DC but showed no difference in stressed and non-stressed mice. However, TNF-alpha production in macrophages of non-stressed mice was decreased. The production of IFN-g, in Con A- and LPS-treated splenic T cells. In contrast IL-5, IL-10, and IL-23 production was high in T cells of stressed mice. T cells treated with norepinephrine and bet2-adrenergic receptor agonist, Fenoterol. Overall, the production of cytokines in stressed and non-stressed mice shows variation that may have roles in enhancing protection or increased lung infection.
{"title":"Analysis of immune response of stressed male mice during Chlamydia muridarum lung infection","authors":"Nathasha Woart","doi":"10.55632/pwvas.v95i2.1004","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.1004","url":null,"abstract":"Chlamydia lung infection caused by Chlamydia trachomatis is a serious lung infection particularly in infants but there are a few studies about lung infection. Chlamydial genital and lung infections in mice is frequently done with Chlamydia muridarum. The purpose of our study was to explore the effect of stress in lung chlamydia infection in male mice. We hypothesize that stress changes the level of cytokine production in monocytes during lung infection. As a The result from these studies include counting of live cells, differentiation of proliferation of bone marrow derived monocytes where the result showed that non-stressed dendritic cells (DCs) WT mice and stressed macrophages (MO) WT mice had sufficient live cells to study. No difference in IL-b production of macrophages of stressed and non-stress mice was observed. Beta2-adrenergic receptor (b2-AR agonist (fenoterol) and antagonist (ICI 118 551) treatment resulted in difference in cytokine production the effect of agonists and antagonists. TNF-alpha production was high in stressed mice compared to non-stress mice for MO. LPS stimulated TNF-alpha production in DC but showed no difference in stressed and non-stressed mice. However, TNF-alpha production in macrophages of non-stressed mice was decreased. The production of IFN-g, in Con A- and LPS-treated splenic T cells. In contrast IL-5, IL-10, and IL-23 production was high in T cells of stressed mice. T cells treated with norepinephrine and bet2-adrenergic receptor agonist, Fenoterol. Overall, the production of cytokines in stressed and non-stressed mice shows variation that may have roles in enhancing protection or increased lung infection.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78355901","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.v95i2.983
Ashlei Kelly
Pseudomonas aeruginosa (Pa) is an opportunistic pathogen that threatens the health, especially in immunocompromised individuals. Pseudomonas aeruginosa survives everywhere, including in a shuttle of spaceflights. Survival and starvation of Pa in water probably increase or decrease the degree of pathogenicity or virulence, but it is not well discovered. To promote our understanding of the mechanisms of Pa long-term survival in water, investigations on survival kinetics and its pathogenicity to mice are under investigation in our laboratory. Data has shown that under starvation conditions in water, Pa can lead to (i) survival for over 15 years in water; (ii) distinct changes in colony morphology, including loss of pigmentation. This study also aimed to determine the pathogenicity of clinical and environmental isolates of Pa in water under starved conditions. We hypothesized that the starvation of Pa isolates in water results in different survival curves, morphological, that may enhance pathogenicity in mice. Mice infected with starved isolates showed a lesser death rate than non-starved isolates. This project compared the susceptibility of BALB/c, C57BL/6J, and DBA/2 J in lung infection. Our data shows the vulnerability of DBA/2J was high compared to BALB/c or C57BL/6J. Hindlimb-unloading increased the death rate in DBA/2J compared to BALB/c strains. Lung infection of different Pa isolates shows a differential death rate, with the highest death rate caused by non-starved isolates in mice. Overall, long-term starvation of Pa isolates exits may lead to a reduced mortality rate of mice.
{"title":"Evaluation of the pathogenicity of Pseudomonas aeruginosa isolates to strains of mice under stressful conditions","authors":"Ashlei Kelly","doi":"10.55632/pwvas.v95i2.983","DOIUrl":"https://doi.org/10.55632/pwvas.v95i2.983","url":null,"abstract":"Pseudomonas aeruginosa (Pa) is an opportunistic pathogen that threatens the health, especially in immunocompromised individuals. Pseudomonas aeruginosa survives everywhere, including in a shuttle of spaceflights. Survival and starvation of Pa in water probably increase or decrease the degree of pathogenicity or virulence, but it is not well discovered. To promote our understanding of the mechanisms of Pa long-term survival in water, investigations on survival kinetics and its pathogenicity to mice are under investigation in our laboratory. Data has shown that under starvation conditions in water, Pa can lead to (i) survival for over 15 years in water; (ii) distinct changes in colony morphology, including loss of pigmentation. This study also aimed to determine the pathogenicity of clinical and environmental isolates of Pa in water under starved conditions. We hypothesized that the starvation of Pa isolates in water results in different survival curves, morphological, that may enhance pathogenicity in mice. Mice infected with starved isolates showed a lesser death rate than non-starved isolates. This project compared the susceptibility of BALB/c, C57BL/6J, and DBA/2 J in lung infection. Our data shows the vulnerability of DBA/2J was high compared to BALB/c or C57BL/6J. Hindlimb-unloading increased the death rate in DBA/2J compared to BALB/c strains. Lung infection of different Pa isolates shows a differential death rate, with the highest death rate caused by non-starved isolates in mice. Overall, long-term starvation of Pa isolates exits may lead to a reduced mortality rate of mice.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"10 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77577891","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.v95i2.990
Weidong Liao, Andrew Moseman
Traditionally, chess engines use handcrafted evaluation functions based on human strategy. Recently, machine learning has been used as an alternative to direct position scoring. However, this typically involves training a model on human matches. Reinforcement learning has been shown to be a viable machine learning approach that, when combined with self play, can train a neural network for chess position evaluation without the need for human domain knowledge. This paper discusses our implementation of a reinforcement learning based chess engine, trained using self play.
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Pub Date : 2023-04-18DOI: 10.55632/pwvas.v95i2.1003
T. Hill
TYLER P. HILL, and HOLLY RACINE, Dept of Biomedical Science, West Liberty University, West Liberty, WV, 26074, Validation of an Avian Model of Induced-Thyrotoxicosis This project aimed to validate the avian model developed in our lab with the ultimate goal of discovering the potential link between maternal hyperthyroidism and craniosynostosis (CS). The method of validation for this model was to qualitatively measure the levels of thyroxine (T4) present in the treated embryos' systems at different time points post-injection. T4 was fluorescently labeled using Alexa fluorophore 488 and was injected into an experimental group of N=37 on embryonic day 11 (E11). Three control groups were used one N=9 Saline injected, another N=5 injected with the unconjugated free dye, and lastly, N=5 T4 injected only. The results showed that after 24 hours post-injection the fluorescent tag was present in the albumen with a peak at 72 hours, after 48 hours fluorescence is detected in the yolk with a peak at 96 hours, finally, after 72 hours there is a large peak in the blood before becoming undetectable at 96 hours. These trends suggest the injection is traveling from the injection site through these tissues and ultimately into the bloodstream, which is supported by the molecular assaying via ELISAs previously conducted. The results of this project in conjunction with the ELISA data show that the peaks detected are from injected T4 rather than a natural spike in development. Prior physiological data collected supports that the T4 injection not only makes its way into the bloodstream but also has a metabolic effect leading to changes in development. These results combined validate this model of induced thyrotoxicosis. (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence) Acknowledgment of NASA West Virginia Space Grant Consortium (Grant #80NSSC20M0055)
TYLER P. HILL和HOLLY RACINE, West Liberty大学生物医学科学系，West Liberty, WV, 26074，诱导性甲状腺毒症鸟类模型的验证本项目旨在验证我们实验室开发的鸟类模型，最终目的是发现母体甲状腺功能亢进与颅骨闭锁(CS)之间的潜在联系。该模型的验证方法是定性测量注射后不同时间点处理胚胎系统中甲状腺素(T4)的水平。用Alexa荧光团488对T4进行荧光标记，于胚胎第11天(E11)注射到N=37的实验组。3个对照组，N=9注射生理盐水，N=5注射未偶联游离染料，N=5只注射T4。结果表明，注射后24小时，蛋白中出现荧光标记，72小时出现荧光峰，48小时后，蛋黄中出现荧光，96小时出现荧光峰，72小时后，血液中出现一个大峰，96小时时无法检测到荧光。这些趋势表明，注射是从注射部位经过这些组织并最终进入血液，这得到了先前通过elisa进行的分子分析的支持。本项目的结果与ELISA数据相结合表明，检测到的峰值来自注射T4，而不是发育过程中的自然峰值。先前收集的生理学数据支持T4注射不仅进入血液，而且具有导致发育变化的代谢作用。这些结果共同验证了这种诱导甲状腺毒症模型。(由NIH资助P20GM103434向西弗吉尼亚IDeA网络提供卓越生物医学研究支持)
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