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Computational insights on the structure of transition states and intermediates of the Boyland-Sims Oxidation reaction Boyland-Sims氧化反应过渡态和中间体结构的计算见解
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.1006
Isaac Hall
The Boyland-Sims Oxidation reaction uses an alkaline solution of peroxydisulfate salt to convert arylamines to o-aminophenols. The long-standing mechanism involves a nucleophilic attack by the amine leading to the formation of an arylhydroxylamine-O-sulfonate intermediate, which then rearranges to the arylamine o-sulfate. The long-standing mechanism of a nucleophilic attack by the amine on peroxide oxygen has been questioned by Marjanović et al. who have proposed a nitrenium ion intermediate as the reactive species rather than the uncharged amine. It has been challenged by a variety of experimental observations. We present here new computations at a high-level of density functional theory (B3LYP/6-311++G**) to model both arylhydroxylamine-O-sulfonate and nitrenium ion pathways in a series of aromatic amines. In this study, the energetics of reaction intermediates in both mechanisms are computed for aniline, 2,4-dinitroaniline, and N,N-dimethylaniline. To further support the pathways of the reaction, the calculations were performed to find the transition states involved in both mechanisms. The transition state calculations revealed two possible energetically feasible pathways for the rearrangement of arylhydroxylamine-O-sulfonate to arylamine o-sulfate. Our results indicate that the Boyland-Sims oxidation reactions proceed via arylhydroxylamine-O-sulfonate intermediate as predicted earlier.
Boyland-Sims氧化反应使用过硫酸氢盐的碱性溶液将芳胺转化为邻氨基酚。长期存在的机制涉及胺的亲核攻击,导致形成芳基羟胺-邻磺酸盐中间体,然后重新排列成芳基胺-邻硫酸酯。marjanoviki等人对胺对过氧的亲核攻击的长期机制提出了质疑,他们提出了一种氮离子中间体而不是不带电的胺作为反应物质。它受到了各种实验观察的挑战。在这里,我们提出了一个新的计算方法,在高密度泛函理论(B3LYP/6-311++G**)的高层次上模拟芳基羟胺-邻磺酸盐和氮离子在一系列芳香胺中的途径。本研究计算了苯胺、2,4-二硝基苯胺和N,N-二甲基苯胺两种反应机制中中间体的能量学。为了进一步支持反应的途径,进行了计算,以找到两种机制中涉及的过渡态。过渡态计算揭示了芳基羟胺-邻磺酸盐重排至邻硫酸芳胺的两种可能的能量可行途径。我们的结果表明,Boyland-Sims氧化反应通过芳基羟胺- o -磺酸盐中间体进行,正如之前预测的那样。
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引用次数: 0
Induced Thyrotoxicosis Elicits Protective Effects in Developing Hearts 诱导甲状腺毒症对发育中的心脏有保护作用
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.958
Kirsten Bogunovich, Holly L. Racine
Maternal hyperthyroidism effects development in infants and can lead to conditions such as craniosynostosis, exophthalmos, and cardiac hypertrophy. Models for studying maternal hyperthyroidism, or thyrotoxicosis, are limited. Our lab has established an avian model to study the effects of induced thyrotoxicosis on embryonic development. Thyroid hormones (TH) cause inotropic and chronotropic changes in cardiac tissue, including increased cardiac output, increased blood volume, and hypertrophy. Therefore, we wanted to investigate the cardiac changes in our model. Fertilized chicken eggs are injected on embryonic days (E) 11 and 15 with either saline (control) or 25ng thyroxine (T4). Hearts were harvested on day E19, with halves of each sample used for histological staining with Masson’s Trichrome and qRT-PCR analysis. The main objective was to observe systemic effects of our model by studying morphological and genetic changes in the heart following exposure by measuring expression of THRa (TR receptor), ATP2A2 (calcium ATPases), and MYH7(Myosin heavy chain 7). We hypothesized that levels of all 3 markers would be upregulated, since THRaregulates transcription of these other cardiac markers due to fluctuating levels of TH. These are linked to ventricular hypertrophy when upregulated. However, our results contradicted our hypothesis. There was downregulation of all 3 genes, with a significant downregulation in ATP2A2. In combination with other data collected in our lab and related literature, we suspect that there is a cardiac protective effect occurring in the hearts following thyroxine exposure.
母亲甲状腺功能亢进影响婴儿的发育,并可导致颅缝闭锁、眼球突出和心脏肥厚等疾病。研究母体甲状腺机能亢进或甲状腺毒症的模型是有限的。本实验室建立了鸟类模型,研究诱导性甲状腺毒症对胚胎发育的影响。甲状腺激素(TH)引起心脏组织的肌力和变时性变化,包括心输出量增加、血容量增加和肥厚。因此,我们想研究我们模型的心脏变化。受精卵在胚胎日(E) 11和15注射生理盐水(对照)或25ng甲状腺素(T4)。在第E19天收获心脏,每个样本的一半用于马松三色和qRT-PCR分析的组织学染色。主要目的是通过测量THRa (TR受体)、ATP2A2(钙atp酶)和MYH7(肌球蛋白重链7)的表达来研究暴露后心脏的形态学和遗传变化,观察我们模型的全身效应。我们假设所有3种标记物的水平都将上调,因为THRa调节其他心脏标记物的转录,这是由于TH水平的波动。当上调时,这些与心室肥厚有关。然而,我们的结果与我们的假设相矛盾。3个基因均下调,其中ATP2A2显著下调。结合我们实验室收集的其他数据和相关文献,我们怀疑甲状腺素暴露后心脏中存在心脏保护作用。
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引用次数: 0
Assessing the interactions of variables affecting the rate of nonhuman decomposition 评估影响非人类分解速率的变量之间的相互作用
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.955
Serenity Parsell, Kristy Henson
Decomposition of organic tissue is a complex process that is affected by a large number of interactions. These variables affect the rate of decomposition, insect activity, and the onset of each individual stage in the decomposition process. Of the variables in effect, some of the most commonly studied are temperature and moisture. The purpose of this study is to analyze the impact of other variables, such as age at death, body size, sun exposure, and presence of preservation chemicals on the rate of decomposition in nonhuman specimens. The goal of this research is to give more insight into the complex interactions between these variables and analyze the reliability of current decomposition analysis. As ethical considerations and policies affect how researchers use human remains, more decomposition research is needed to better understand this very sensitive process and all the variables that are included. Preliminary results suggest that sun exposure, relative body size, and age at death do not significantly affect the rate of nonhuman decomposition, but a principal component analysis suggests correlation does exist between subsets of these variables. Ongoing work to include larger sample sizes, as well as chemically preserved animals, is currently running. Expected results indicate that preserved animals will take longer to fully decompose, and the effects of age, sun exposure, and body size will not be significant.  
有机组织的分解是一个复杂的过程,受到大量相互作用的影响。这些变量影响分解速率、昆虫活动和分解过程中每个单独阶段的开始。在起作用的变量中,最常被研究的是温度和湿度。本研究的目的是分析其他变量的影响,如死亡年龄、体型、日晒和保存化学品的存在对非人类标本分解速度的影响。本研究的目的是更深入地了解这些变量之间复杂的相互作用,并分析当前分解分析的可靠性。由于伦理考虑和政策会影响研究人员如何使用人类遗体,因此需要进行更多的分解研究,以更好地了解这一非常敏感的过程以及包含的所有变量。初步结果表明,日晒、相对体型和死亡年龄对非人类尸体的分解率没有显著影响,但主成分分析表明,这些变量的子集之间确实存在相关性。目前正在进行的工作包括更大的样本量,以及化学保存的动物。预期的结果表明,保存的动物需要更长的时间才能完全分解,年龄、日晒和体型的影响并不显著。
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引用次数: 0
The analyzation of DNA and cross contamination inside the cyanoacrylate (superglue) fuming chamber. 氰基丙烯酸酯(强力胶)发烟室内DNA及交叉污染分析。
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.981
Casie Banyi
The objectives of this experiment looked at were that the DNA, both skin cells and saliva, have the potential of binding with the superglue as it coats each object being fumed. If DNA has the potential of binding with the superglue, then it is very possible for DNA to be carried in the air by the humidifier, thus contaminating other objects. For the first experiment, two saliva covered microscope slides, one female and one male, were fumed along with 25 other sterile slides. Each sterile slide was swabbed and analyzed for DNA in an electrophoresis gel chamber. The second experiment fumed ten microscope slides presented with fingerprints. Each sterile slide was swabbed and analyzed for DNA in an electrophoresis gel chamber. The results showed that there was minimal cross contamination present on the slides fumed with the saliva based slides. The touch DNA experiment showed that there are potentially identifiable DNA markers present in fingerprints after fuming. The results concluded that it is possible to have DNA cross contamination between items that carry no DNA prior to the fumigation, and potential DNA can be found on fingerprints after fumigation.
这个实验的目的是观察DNA,包括皮肤细胞和唾液,当它覆盖在每个被熏制的物体上时,都有与超级胶水结合的潜力。如果DNA具有与强力胶结合的潜力,那么DNA极有可能被加湿器携带在空气中,从而污染其他物体。在第一个实验中,两张唾液覆盖的显微镜载玻片,一张是女性,一张是男性,与其他25张无菌载玻片一起被熏制。每张无菌载玻片拭子并在电泳凝胶室中分析DNA。第二个实验用烟熏了十张带有指纹的显微镜载玻片。每张无菌载玻片拭子并在电泳凝胶室中分析DNA。结果表明,在以唾液为基础的载玻片上存在最小的交叉污染。触摸DNA实验表明,熏烟后指纹中存在潜在可识别的DNA标记。结果表明,熏蒸前不携带DNA的物品之间可能存在DNA交叉污染,熏蒸后指纹上可能存在DNA。
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引用次数: 0
The somatosensory and visual systems adaptation times have a strong positive correlation.   体感系统和视觉系统的适应时间有很强的正相关关系。
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.950
Natalia Omelchenko-Comer
CONNIE BURRIS, MARIA MARTINEZ, RAE MOORE, SARAH ZAMZOW, NATALIA OMELCHENKO-COMER, STEM and Business Division, WVNCC, Wheeling, WV, 26003. The somatosensory and visual systems adaptation times have a strong positive correlation.   Multiple studies demonstrated that all sensory systems adapt their sensitivity levels to improve their responsiveness to changes in corresponding stimulation and to accelerate following CNS processing.  The study aimed to determine if the time of adaptation observed for the somatosensory system correlates with the adaptation time observed for the visual system. The data collection was performed on 2 male and 7 female healthy volunteers (the average age of those was 22.2+/-4.60, 8 right-handed and 1 left-handed) in a seated position. The adaptation time for the somatosensory system was estimated by recording the time of the perception loss after placing a 1 penny coin onto the dorsal hand surface. To evaluate the adaptation time for the visual system the subject was asked to stare at a small pastel color abstract image. The time of the perceived loss for the component of this image was recorded. The average adaptation time for somatosensory perception was 22.0+/-16.15 seconds ranging from 4 to 42 seconds. The average adaptation time for visual perception was 30.7+/-16.04 seconds ranging from 12 to 51 seconds. There was a statistically significant difference between the adaptation time for these modalities of the sensations as determined by paired T-test (t (8) = 3.1, p = .01).  However, the correlation analysis demonstrated a strong positive correlation between values (r (8) = 0.86, p = .003). Our data indicate that the times of somatosensory and visual adaptation are strongly correlated. The finding of this study can be used for the development and improvement of neurobehavioral evaluation tests.
康妮·伯利斯,玛丽亚·马丁内斯,雷·摩尔,萨拉·赞佐,娜塔莉亚·奥梅尔肯科,STEM和业务部门,WVNCC,惠灵,西弗吉尼亚州,26003。体感系统和视觉系统的适应时间有很强的正相关关系。多项研究表明,所有的感觉系统都可以调整它们的敏感性水平,以提高它们对相应刺激变化的反应能力,并加速中枢神经系统的后续处理。该研究旨在确定体感系统的适应时间是否与视觉系统的适应时间相关。2名男性和7名女性健康志愿者(平均年龄22.2±4.60岁,8名右撇子和1名左撇子)以坐姿进行数据收集。躯体感觉系统的适应时间是通过记录将1便士硬币放在手背表面后感知丧失的时间来估计的。为了评估视觉系统的适应时间,被试被要求盯着一个小的粉彩抽象图像。记录该图像分量的感知损失时间。体感知觉的平均适应时间为22.0+/-16.15秒,范围为4 ~ 42秒。视觉感知的平均适应时间为30.7±16.04秒,范围为12 ~ 51秒。配对t检验表明,两种感觉模式的适应时间差异有统计学意义(t (8) = 3.1, p = 0.01)。然而,相关分析显示值之间存在很强的正相关(r (8) = 0.86, p = 0.003)。我们的数据表明,体感和视觉适应的时间是密切相关的。本研究结果可为神经行为评估测试的发展和改进提供参考。
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引用次数: 0
Effects of Handwashing and Antimicrobials on Bacterial Counts 洗手和抗菌剂对细菌计数的影响
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.949
Emily Hissom
It is everyday practice to utilize aseptic techniques, which are simply actions taken to prevent the accumulation of bacteria and other microbes to prevent diseases and their spread. Handwashing is an important aspect of hand hygiene because hands pick up the microbes from surfaces and are easily transported from person to person through direct contact or indirect contact. According to the CDC and WHO, many diseases are spread by not washing hands effectively, therefore hand hygiene is important everywhere in society, especially in healthcare where the spread of disease can mean life or death. In this study, we analyzed data collected from the student population of microbiology classesfrom the University of Charleston in West Virginia over the span of nine years (2014 to present). We measured the number of colony forming units (CFU) from students’ hands prior to manipulation (control), using just water, using soap and water, and using hand sanitizer. We hypothesized that handwashing and use of antimicrobial aids such as hand sanitizer will reduce the number of bacteria on hands compared to controls. An analysis was also performed comparing pre-pandemic and post-pandemic data to determine if handwashing behaviors have changed behavior which would alter the normal microfauna on the hands. This analysis revealed that there was a significant difference between handwashing techniques (p<0.001). There was a difference in formation of colony forming units between pre and post pandemic times (p<0.001).
使用无菌技术是日常的做法,这只是采取的行动,以防止细菌和其他微生物的积累,以防止疾病及其传播。洗手是手卫生的一个重要方面,因为手从表面携带微生物,并且很容易通过直接接触或间接接触在人与人之间传播。根据疾病预防控制中心和世界卫生组织的说法,许多疾病都是由于不洗手而传播的,因此洗手卫生在社会的任何地方都很重要,特别是在疾病传播可能意味着生命或死亡的医疗保健领域。在这项研究中,我们分析了从西弗吉尼亚州查尔斯顿大学微生物课的学生群体中收集的9年(2014年至今)的数据。在操作(对照组)前,我们测量了学生手上的菌落形成单位(CFU)的数量,使用清水,使用肥皂和水,以及使用洗手液。我们假设,与对照组相比,洗手和使用抗菌辅助工具(如洗手液)可以减少手上的细菌数量。还进行了一项分析,比较大流行前和大流行后的数据,以确定洗手行为是否改变了行为,从而改变了手上正常的微动物群。该分析显示,洗手方式之间存在显著差异(p<0.001)。大流行前后菌落形成单位的形成存在差异(p<0.001)。
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引用次数: 0
The S-STEM Scholarship Program to Advance Academic Success and Career Development for CME Majors at Shepherd University S-STEM奖学金项目旨在促进谢泼德大学CME专业学生的学业成功和职业发展
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.1007
Qing Wang, Amy L. Dewitt, Emily Gross, Weidong Liao, K. Adams, R. Mirdamadi, Osman Guzide
The Track II NSF S-STEM Program in the Computer Sciences, Mathematics, and Engineering Department at Shepherd University started in October 2021 and has thus far supported 14 scholars majoring in computer science, mathematics, data analytics, or engineering. The overall goal of the program is to increase the number and quality of STEM graduates in the STEM workforce by providing financial, academic, and social support, which enables qualified students to overcome certain barriers that prevent them from meeting their educational and career goals. In this poster, the most significant activities and outcomes of the program since its inception are presented. Survey results and outcomes from the last academic year, along with initiatives that increase, retain, and support students in CME fields, are also discussed. This program has been supported by the NSF S-STEM Grant (award No. DUE-2130267).  
谢泼德大学计算机科学、数学和工程系的第二轨NSF S-STEM项目于2021年10月启动,迄今已资助了14名计算机科学、数学、数据分析或工程专业的学者。该计划的总体目标是通过提供经济、学术和社会支持,提高STEM劳动力中STEM毕业生的数量和质量,使合格的学生能够克服阻碍他们实现教育和职业目标的某些障碍。在这张海报中,介绍了该计划自成立以来最重要的活动和成果。本文还讨论了上一学年的调查结果和结果,以及增加、留住和支持CME领域学生的举措。本项目由美国国家科学基金S-STEM基金(编号:No. 5)资助。由于- 2130267)。
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引用次数: 0
Taxonomic Standing and Genetic Surveillance of the Spiny Scale Crayfish (Cambarus jezerinaci) in Southeastern Kentucky 肯塔基州东南部刺鳞螯虾的分类地位和遗传监测
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.986
Cassidy Gebhardt, Zachary J. Loughman
  In Southeastern Kentucky a small, cold stream-dwelling crayfish, Cambarus jezerinaci, located in the Kentucky and Cumberland River basin may be a part of a species complex. A previous study done in 2008 on the cytochrome oxidase 1 gene (CO1) of C. jezerinaci and Cambarus parvoculus, a sister species in the region, identified a third genetically distinct lineage most similar to C. jezerinaci currently retained as Group B C. jezerinaci. Group B is as different from the true species as the true species is to C. parvoculus, a total CO1 difference greater than 5%.  Morphological characteristics were also compared between C. jezerinaci and C. parvoculus with only rostral states being significant to species identification. However, in light of new genetic lineages previous research suggested the separation of them before morphological comparisons. In 2022 and 2023 research was conducted on both the genetics and morphological differences of C. jezerinaci within two river basins to better understand the taxonomic standing of this crayfish. Determining the true status of C. jezerinaci within these two river basins is important to the conservation of these animals. Continuing genetic research helps us to grow our understanding of what it means to be a species in the modern age and also allows us to learn about the biodiversity and speciation process itself. Given the rapidly changing climate, it is imperative to catalog the true diversity of our freshwater streams and develop conservation action plans for endemic species such as C. jezerinaci.
在肯塔基州东南部,一种生活在肯塔基州和坎伯兰河流域的小型冷溪小龙虾,Cambarus jezerinaci,可能是一个物种综合体的一部分。之前在2008年对该地区的姐妹种——C. jezerinaci和Cambarus parvoculus的细胞色素氧化酶1基因(CO1)进行的一项研究,发现了第三个遗传上独特的谱系,与目前保留为B群的C. jezerinaci最相似。B组与真种的差异与真种与parvoculus的差异一样大,总co2差异大于5%。并比较了jezerinaci和parvoculus的形态特征,其中只有吻侧状态对物种鉴定有重要意义。然而,鉴于新的遗传谱系,先前的研究表明,在形态学比较之前,它们是分开的。为了更好地了解这种小龙虾的分类地位,研究人员于2022年和2023年对两河流域的切泽里纳螯虾进行了遗传和形态差异研究。确定这两个河流流域的真实状况对这些动物的保护具有重要意义。持续的基因研究有助于我们加深对现代物种意义的理解,也让我们了解生物多样性和物种形成过程本身。考虑到气候的迅速变化,对我们淡水溪流的真正多样性进行编目并制定地方性物种(如C. jezerinaci)的保护行动计划势在必行。
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引用次数: 0
Superposition as a Means of Data Encryption in N-Dimensional Value Spaces n维值空间中数据加密的叠加方法
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.956
Tyler Burgee
The objective of this study was to provide a new method for creating quantum-proof encryption algorithms. I accomplished this by designing a symmetric 2-key cryptosystem that exploits the superposition principle to encrypt data in multi-dimensional value spaces.    The proposed cryptosystem substitutes characters for frequencies, as determined by two private keys: component wave order key (CWOK) and character transmission order key (CTOK). A CWOK defines the values and theoretical spatial arrangement of frequencies in a complex wave. A CTOK defines the unique arrangement of system characters (i.e., characters in an encoding scheme such as ASCII), determined by a hash function, to identify a user. Combining the CWOK and CTOK, we construct a character-lookup table (CLT), which defines the character-frequency relationships used to generate a substitution cipher. A cipher’s frequency values must be superimposed in accordance with the CWOK. Fast Fourier Transforms are used during the decryption stage to perform complex wave analysis.    Complex waves can have n! frequency configurations, where n = the number of component frequencies; each CTOK can have a! character configurations, where a = the number of characters defined in an encoding scheme. Therefore, by requiring n ≥128 and using the ASCII encoding scheme (a = 128), there are n!+a!=128!+128!=2*128! possible key configurations for any given cipher. This is approximately 3.330284e+138 times as many key configurations possible with AES 256.    Exploiting the multi-dimensional nature of complex waves, and combining these techniques with other powerful encryption algorithms used today, it appears likely that we can create a quantum-proof cryptosystem.
本研究的目的是提供一种创建量子证明加密算法的新方法。我通过设计一个对称的2密钥密码系统来实现这一点,该系统利用叠加原理对多维值空间中的数据进行加密。提出的密码系统用字符代替频率,由两个私钥确定:分量波序密钥(CWOK)和字符传输顺序密钥(CTOK)。CWOK定义了复波中频率的值和理论空间排列。CTOK定义了由哈希函数确定的系统字符(即编码方案(如ASCII)中的字符)的唯一排列,以标识用户。结合CWOK和CTOK,我们构建了一个字符查找表(CLT),它定义了用于生成替换密码的字符-频率关系。密码的频率值必须按照CWOK进行叠加。在解密阶段使用快速傅里叶变换来执行复波分析。复波可以有n!频率配置,其中n =组成频率的个数;每个CTOK都可以有一个!字符配置,其中a =在编码方案中定义的字符数。因此,通过要求n≥128,并使用ASCII编码方案(a =128),有n!+a!=128!+128!=2*128!任何给定密码的可能密钥配置。这大约是AES 256可能的密钥配置的3.330284e+138倍。利用复杂波的多维特性,并将这些技术与目前使用的其他强大的加密算法相结合,我们似乎可以创建一个量子证明的密码系统。
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引用次数: 0
Applying RNA-Seq to investigate the transition into a Viable But Non Culturable State (VBNC) for the intracellular pathogen, Francisella tularensis LVS. 应用RNA-Seq技术研究土拉弗朗西斯菌(Francisella tularensis LVS)胞内病原菌向活但不可培养状态(Viable But Non - culable State, VBNC)转变的过程。
Pub Date : 2023-04-18 DOI: 10.55632/pwvas.v95i2.994
S. Cantlay, Nicole L. Garrison, Rachelle Patterson, D. Primerano, Jun Fan, Joseph Horzempa
Many species of bacteria, under conditions of stress or nutrient limitation, enter a state of dormancy referred to as viable but non-culturable (VBNC). VBNC bacteria persist in the environment, are difficult to detect and identify by many standard laboratory methods and can be altered in their susceptibility to antibiotics. Entry into the VBNC state is often accompanied by morphological changes; however, the mechanisms underlying this are poorly understood. Francisella transitions rapidly and spontaneously to a VBNC state, and therefore has the potential to be an excellent model organism for the study of this phenomenon. To investigate the transcriptome of VBNC F. tularensis LVS we extracted RNA from culturable and VBNC cells and carried out an RNA-Seq analysis using both Long-read Nanopore and Illumina Sequencing. Differentially expressed genes (DEGs) were identified using a DE-Seq pipeline. Over 300 genes were significantly upregulated and ~100 genes were down regulated in VBNC cells. Amongst the upregulated genes were some involved in the transport of metals or small molecules, and we have also identified putative transcriptional regulators that may be master controllers of the VBNC process. Our data represents the first transcriptomic analysis of F. tularensis LVS as it transitions into the VBNC state. Identifying genes that are involved in this transition is a critical first step in understanding the mechanisms that drive F. tularensis LVS into the VBNC state and will help us identify factors that allow the resuscitation of these bacteria. This will have important implications for understanding  both environmental persistence and pathogenicity of Francisella species. (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence)
许多种类的细菌,在应激或营养限制的条件下,进入一种休眠状态,称为可活但不可培养(VBNC)。VBNC细菌在环境中持续存在,难以通过许多标准实验室方法检测和鉴定,并且可以改变其对抗生素的敏感性。进入VBNC状态往往伴随着形态变化;然而,人们对其背后的机制知之甚少。Francisella可以快速自发地过渡到VBNC状态,因此有可能成为研究这一现象的优秀模式生物。为了研究VBNC F. tularensis LVS的转录组,我们从可培养细胞和VBNC细胞中提取RNA,并使用长读纳米孔和Illumina测序技术进行RNA- seq分析。使用DE-Seq管道鉴定差异表达基因(DEGs)。在VBNC细胞中,超过300个基因显著上调,约100个基因显著下调。在上调的基因中,有一些涉及金属或小分子的运输,我们还确定了可能是VBNC过程主控者的转录调控因子。我们的数据代表了土拉菌LVS转变为VBNC状态时的首次转录组学分析。识别参与这种转变的基因是理解驱动土拉菌LVS进入VBNC状态的机制的关键的第一步,并将帮助我们确定允许这些细菌复苏的因素。这将对了解弗朗西斯菌的环境持久性和致病性具有重要意义。(NIH拨款P20GM103434资助西弗吉尼亚IDeA网络生物医学卓越研究)
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引用次数: 0
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Proceedings of the West Virginia Academy of Science
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