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The Synthesis of Protein Polymer Conjugates using the Human Regulatory Protein Galectin-3 利用人调节蛋白半乳糖凝集素-3合成蛋白质聚合物偶联物
Pub Date : 2020-02-13 DOI: 10.32473/ufjur.v21i2.108726
Ling Lin, Amanda M. Pritzlaff, H. C. Lower, D. Savin
Galectin-3 (gal3) is a human lectin protein that is known to interact with extracellular matrix proteins by regulating functions in both healthy and cancerous cells. The goal of this project is to conjugate polymers to gal3 to better study and control its functions in vitro. We hypothesize that a covalently attached polymer will sterically modulate gal3 function. In the project, we created two protein variants with polymer-reactive handles. The first construct is similar to wild-type gal3 with a cysteine in place of the 6th serine (S6C) which was created by site-directed mutagenesis (SDM). Maleimide-terminated polyethylene oxide (PEO, 5000 g/mol) was then attached to this mutant via thiol-Michael addition at the cysteine site. Attachment of polymer to the unstructured N-terminal domain (NTD) may increase the binding of the protein by sterically pulling the NTD away from the carbohydrate recognition domain (CRD). In addition, the NTD, which is implicated in undesired self-association, was removed for the second construct. The gal3 CRD only construct is shown to have a higher solubility in solution and an increased ligand-binding affinity. Ultimately, the two unique constructs will help us understand the structural role of the NTD in gal3 ligand-binding and self-association.
半乳糖凝集素-3 (gal3)是一种人类凝集素蛋白,已知通过调节健康细胞和癌细胞的功能与细胞外基质蛋白相互作用。本项目的目的是将聚合物与gal3偶联,以便更好地研究和控制其体外功能。我们假设共价连接的聚合物将立体调节gal3的功能。在这个项目中,我们创建了两个具有聚合物反应手柄的蛋白质变体。第一个结构类似于野生型gal3,用半胱氨酸代替由位点定向突变(SDM)产生的6号丝氨酸(S6C)。然后在半胱氨酸位点通过巯基- michael加成将端部为马来酰亚胺的聚乙烯氧化物(PEO, 5000 g/mol)连接到该突变体上。聚合物附着在非结构n端结构域(NTD)上,通过将NTD从碳水化合物识别结构域(CRD)上拉离,增加了蛋白质的结合。此外,NTD在第二个构念中被移除,它与不希望的自我联想有关。仅gal3 CRD结构体在溶液中具有更高的溶解度和更高的配体结合亲和力。最终,这两个独特的结构将帮助我们理解NTD在gal3配体结合和自结合中的结构作用。
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引用次数: 0
Role of a Specific P53 Binding Site in Limiting Tissue Overgrowth 特定P53结合位点在限制组织过度生长中的作用
Pub Date : 2020-02-13 DOI: 10.32473/ufjur.v21i2.108518
Cheyenne Heflin, Haya Ghannouma, Lei Zhou
The p53 protein is an important transcription factor known for maintaining tissue homeostasis by activating genes that have antiproliferative function, such as pro-apoptotic and cytostatic genes. Transcriptional activation of proapoptotic genes has displayed a fundamental role in mediating apoptosis during Drosophila development. Using ChIP-Seq and RNA-Seq methods, we have identified p53 binding sites potentially responsible for p53-mediated induction of pro-apoptotic genes following DNA damage. We have since generated fly lines with the p53 binding site deleted by CRISPR-Cas9-mediated genome editing. To study the effects of the p53 binding site knockout (p53BSKO) on tissue homeostasis, wings of the knockout fly line were dissected, mounted, and then compared against WT wings. Results show p53BSKO animals had an increase in wing size compared to that of the WT. FijiWings 2.2 macros software was used to measure wing hair (trichome) densities, which is directly proportional to cell numbers. This analysis showed that p53BSKO led to hyperplasia of the wing as compared to the WT. Our study indicated that this single P53BS is required for ensuring the right number of cells in a given tissue, likely through mediating overproliferation-induced apoptosis.
p53蛋白是一种重要的转录因子,通过激活具有抗增殖功能的基因(如促凋亡基因和细胞抑制基因)来维持组织稳态。在果蝇发育过程中,促凋亡基因的转录激活在介导细胞凋亡中发挥了重要作用。使用ChIP-Seq和RNA-Seq方法,我们已经确定了p53结合位点,可能负责p53介导的DNA损伤后促凋亡基因的诱导。此后,我们通过crispr - cas9介导的基因组编辑产生了p53结合位点被删除的果蝇系。为了研究p53结合位点敲除(p53BSKO)对组织稳态的影响,我们解剖了敲除果蝇系的翅膀,并将其与WT的翅膀进行了比较。结果显示,p53BSKO动物的翅膀尺寸比WT增加。FijiWings 2.2宏软件测量了翼毛(毛状体)密度,毛状体密度与细胞数量成正比。该分析表明,与WT相比,p53BSKO导致了翅膀的增生。我们的研究表明,单一的P53BS是确保给定组织中正确数量的细胞所必需的,可能是通过介导过度增殖诱导的细胞凋亡。
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引用次数: 0
Role of p53-Mediated Apoptosis in Limiting Metastasis p53介导的细胞凋亡在限制转移中的作用
Pub Date : 2020-02-13 DOI: 10.32473/ufjur.v21i2.108535
Sandra Anne Hegeman, Haya Ghannouma, Lei Zhou
p53 is a transcription factor known to play important roles in limiting tumorigenesis, including controlling the induction of pro-apoptotic genes and apoptosis in response to oncogenic stress. Using ChIP-Seq and RNA-Seq, our lab has identified p53 binding sites potentially responsible for p53-mediated induction of pro-apoptotic genes following DNA damage. Using CRISPR Cas9, we have generated fly strains with deletions or mutations within the p53 binding motif located in the experimentally verified p53 binding site. This deletion blocked DNA damage induced apoptosis. To study the functionality of the motif in limiting tumorigenesis, we introduced the deletion into a genetic tumorigenesis and metastasis model, where the tumor-suppressor cell polarity gene scribble was knocked down via shRNAi in the non-essential tissue compound eye and introduced together with strong oncogenic mutation that can lead to neoplasia and metastasis. We developed a scoring scale to quantify the severity of neoplasia and metastasis, where 0 indicates a wild type phenotype and 5 indicates the most severe metastatic phenotype. Results show that the p53BSKO predominantly displayed a score of 5 (39%). However, our preliminary results also suggested that the severity of this model may also subject to genetic background besides the p53 binding motif.
P53是一种已知在限制肿瘤发生中发挥重要作用的转录因子,包括控制促凋亡基因的诱导和在致癌应激下的细胞凋亡。使用ChIP-Seq和RNA-Seq,我们的实验室已经确定了p53结合位点,可能负责p53介导的DNA损伤后促凋亡基因的诱导。利用CRISPR Cas9,我们在实验验证的p53结合位点上产生了p53结合基序缺失或突变的蝇株。这种缺失阻断了DNA损伤引起的细胞凋亡。为了研究该基序在限制肿瘤发生方面的功能,我们将其缺失引入到遗传肿瘤发生和转移模型中,在非必需组织复眼中通过shRNAi敲除肿瘤抑制细胞极性基因scribble,并将其与可导致肿瘤发生和转移的强致癌突变一起引入。我们制定了一个评分量表来量化肿瘤和转移的严重程度,其中0表示野生型表型,5表示最严重的转移表型。结果显示,p53BSKO以5分为主(39%)。然而,我们的初步结果还表明,除了p53结合基序外,该模型的严重程度还可能受遗传背景的影响。
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引用次数: 1
Perception and Experience of In-Sequence and Out-of-Sequence Engineering Students in a General Chemistry Laboratory 普通化学实验室中顺序与无序工程学生的感知与体验
Pub Date : 2020-02-11 DOI: 10.32473/ufjur.v21i2.108496
Courtney Mercedes Spillman, Lorelie Imperial, Kent J. Crippen
The ChANgE Chem (NSF-1625378) utilizes Cognitive Apprenticeship as a theoretical framework for integrating engineering practices into a freshman chemistry laboratory course for engineering majors with the goal of better supporting all students to degree completion. The activities are structured as three-week Design Challenges (DCs) where students use chemistry knowledge to solve authentic engineering problems. This study explores the experiences of students taking the course in-sequence (i.e. fall of freshman year) versus those taking it out-of-sequence (i.e. spring), where out-of-sequence students have been identified as at higher academic risk. Data was collected through audio and video recordings and post-laboratory surveys. Video recordings were coded using a protocol to identify type and frequency of issues and questions asked. The post-laboratory surveys obtained information concerning students’ perception of task difficulty and their feelings of being like an engineer. The data demonstrated that while out-of-sequence students ask more questions and experience more issues, they did feel like successful engineers and did not find the tasks too difficult. Therefore, additional curriculum supports as well as assistance from a Teaching Assistant are needed in order to positively influence the persistence of out-of-sequence students in spite of the challenges they may face.
ChANgE Chem (NSF-1625378)采用认知学徒制作为理论框架,将工程实践整合到工程专业大一化学实验课程中,目的是更好地支持所有学生完成学位。这些活动被安排为为期三周的设计挑战(dc),学生们利用化学知识来解决真正的工程问题。本研究探讨了按顺序上课的学生(即大一秋季)与按顺序上课的学生(即春季)的经历,其中按顺序上课的学生被认为具有更高的学术风险。通过录音、录像和实验后调查收集数据。视频记录使用协议编码,以确定问题和问题的类型和频率。实验后的调查获得了学生对任务难度的感知和他们像工程师一样的感觉的信息。数据表明,虽然乱序的学生提出了更多的问题,经历了更多的问题,但他们确实觉得自己是成功的工程师,并不觉得任务太难。因此,需要额外的课程支持以及助教的协助,以便对无序学生的坚持产生积极影响,尽管他们可能面临挑战。
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引用次数: 0
The Effects of Histone Deacetylase (HDAC) Inhibitors on FASN Intracellular Localization in Cancer Cells 组蛋白去乙酰化酶(HDAC)抑制剂对癌细胞中FASN细胞内定位的影响
Pub Date : 2020-02-10 DOI: 10.32473/ufjur.v21i2.108423
E. Colvin, Daiqing Liao
Understanding mechanisms underlying cancer biology is crucial for discovering novel and effective therapies to improve patient outcome. Increased lipid production is a major metabolic feature in cancer. The fatty acid synthase (FASN) is a key enzyme for lipid synthesis and is upregulated in cancer. Although fatty acid synthesis is generally thought to take place in the cytoplasm, it has been reported that this enzyme also localizes to the nucleus in cancer cells. We hypothesize that the intracellular localization FASN could be a potential target to decrease lipid synthesis and ultimately halt cell proliferation. Protein acetylation has been shown to regulate protein intracellular localization. We aim to assess the impact histone deacetylase inhibitors (HDACi) have on the intracellular location of FASN with the goal of reducing de novo lipid production and cancer cell proliferation. We have examined intracellular localization of FASN in cells using immunofluorescence microscopy in cancer cells treated with HDACi and did not detect obvious HDACi-induced changes in FASN localization. FASN is also regulated by other mechanisms such as phosphorylation. Future studies will examine effects of kinase inhibitors on FASN intracellular localization.
了解潜在的癌症生物学机制对于发现新的和有效的治疗方法来改善患者的预后至关重要。脂质生成增加是癌症的主要代谢特征。脂肪酸合成酶(FASN)是脂质合成的关键酶,在癌症中表达上调。虽然脂肪酸合成通常被认为是在细胞质中进行的,但有报道称这种酶也定位于癌细胞的细胞核中。我们假设细胞内定位FASN可能是减少脂质合成并最终停止细胞增殖的潜在靶标。蛋白质乙酰化已被证明可以调节蛋白质在细胞内的定位。我们旨在评估组蛋白去乙酰化酶抑制剂(HDACi)对FASN细胞内定位的影响,目的是减少新生脂质产生和癌细胞增殖。我们使用免疫荧光显微镜检查了HDACi处理的癌细胞中FASN的细胞内定位,并没有发现HDACi诱导的FASN定位的明显变化。FASN也受其他机制的调控,如磷酸化。未来的研究将研究激酶抑制剂对FASN细胞内定位的影响。
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引用次数: 0
Social Networks and Mental Health of Rural Latino Adolescents in North Florida 北佛罗里达农村拉丁裔青少年的社会网络与心理健康
Pub Date : 2020-02-10 DOI: 10.32473/ufjur.v21i2.108860
K. J. Elliott, S. Dupre, Faan Jeanne-Marie R. Stacciarini Rn
AbstractThere is limited research on mental health in rural Latino communities, particularly with adolescents. The aims of this study were to: 1) describe the evidence of social networks and mental health research in rural Latino communities and 2) illustrate themes related to the social networks and mental health depicted by rural Latino adolescents. A secondary data analysis study was performed, using data collected through semi-structured interviews (N=59) with adolescents, 11-17 years-old, living in three rural counties of North Florida. The interviews were conducted by community health workers (promotoras), in either Spanish or English; exploring family and community relations. NVivo 12 was used to aid in managing and thematically analyzing the data.  Major themes were analyzed as Protective and Risk factors for mental health. A social network framework was then created under these themes including Family, School, Outside Community and Immigration Policy as main categories. The findings of this study highlight the need for mental health promotion interventions with rural Latino adolescents and can be used to influence future studies of social networks and mental health.
农村拉丁裔社区,特别是青少年的心理健康研究有限。本研究的目的是:1)描述农村拉丁裔社区的社会网络和心理健康研究的证据;2)阐明农村拉丁裔青少年的社会网络和心理健康的相关主题。第二项数据分析研究采用半结构化访谈收集的数据(N=59),调查对象为居住在北佛罗里达州三个农村县的11-17岁青少年。访谈由社区卫生工作者(促进者)用西班牙语或英语进行;探索家庭和社区关系。使用NVivo 12来帮助管理和专题分析数据。主要主题作为心理健康的保护因素和危险因素进行分析。然后在这些主题下创建了一个社会网络框架,包括家庭、学校、外部社区和移民政策作为主要类别。这项研究的结果强调了对农村拉丁裔青少年进行心理健康促进干预的必要性,并可用于影响未来社会网络和心理健康的研究。
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引用次数: 1
Learning Dance: Defining Efficiency and Precision Through Animation 学习舞蹈:通过动画定义效率和精度
Pub Date : 2020-02-05 DOI: 10.32473/ufjur.v21i2.108429
Danielle Sara Frost
Bodily efficiency and precision are crucial components in the execution of ballet technique. In conjunction with traditional studio ballet training, three-dimensional animation can showcase correct body placement, posture, and alignment during the execution of ballet technique helping dancers prevent injuries as well as maintain body longevity. Having digital three-dimensional animation as a pedagogical tool can help teachers, students, and parents see how steps should be executed from different angles. Such technology can also bring students’ attention to common mistakes without the risk of repeated human error. Dancers can compare videos of three-dimensional animated figures showcasing correct ballet technique to videos of themselves in order to see where they may be misaligning their bodies and causing potential injuries or bad habits. This pedagogical tool can also be helpful for those who have limited access to taking ballet classes but would still like to learn in an efficient and safe manner. 
身体的效率和精度是执行芭蕾技术的关键组成部分。结合传统的工作室芭蕾舞训练,三维动画可以展示正确的身体位置,姿势和对齐在执行芭蕾舞技术,帮助舞者防止受伤,并保持身体寿命。使用数字三维动画作为教学工具可以帮助老师、学生和家长从不同的角度看到应该如何执行步骤。这种技术还可以让学生注意到常见的错误,而不会有重复人为错误的风险。舞者可以将展示正确芭蕾舞技巧的三维动画人物视频与他们自己的视频进行比较,以便发现他们可能在哪里不对姿势,从而造成潜在的伤害或不良习惯。这个教学工具也可以帮助那些有机会参加芭蕾课的人,但仍然想以一种有效和安全的方式学习。
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引用次数: 0
Gastrointestinal Symptoms in Autism Spectrum Disorders 自闭症谱系障碍的胃肠道症状
Pub Date : 2019-12-05 DOI: 10.32473/ufjur.v21i1.108746
Jenna Logan Schuppener, J. Elder
The purpose of this study was to characterize individuals with autism spectrum disorders (ASD), who are part of a larger ASD study, in order to determine clinically useful ways to evaluate gastrointestinal (GI) symptomology. Thirty-three children (ages 3-16) were recruited from a variety of community agencies. Potential participants were screened using the Autism Diagnostic Observation Schedule (ADOS). Parental informed consent was obtained from all participants and child assent when appropriate. Parents completed the Gastrointestinal Symptom Rating Scale, Bristol Stool Chart (BSC), and sociodemographic form. Of the 33 participants, 14 were currently experiencing gastrointestinal symptoms and had abnormal bowel movements. Seven had a history of gastrointestinal symptoms and abnormal bowel movements; two parents were unsure of their gastrointestinal health. According to the BSC rescore, 13 participants reported hard stools, 14 had normal stools, and six had loose stools. Moderate correlations were found among many GI symptoms (r33= .396 - .751) indicating that these could be clustered to create a profile. These findings are clinically significant because GI symptoms in ASD are difficult to diagnose due to communication and social impairments and may go undiagnosed and untreated.  Further characterizing GI symptomology in this population is a logical role for nurse researchers since nurses are often on the front line intervening within the ASD population.  
本研究的目的是描述自闭症谱系障碍(ASD)患者的特征,以确定临床有用的方法来评估胃肠道(GI)症状,这是一个更大的ASD研究的一部分。从不同的社区机构招募了33名儿童(3-16岁)。使用自闭症诊断观察表(ADOS)筛选潜在的参与者。所有参与者都获得了家长的知情同意,适当时也获得了儿童的同意。家长完成胃肠症状评定量表、布里斯托大便量表(BSC)和社会人口统计表格。在33名参与者中,有14人目前出现胃肠道症状,排便异常。7例有胃肠道症状和排便异常史;两对父母不确定他们的肠胃健康状况。根据BSC评分,13名参与者报告大便坚硬,14名大便正常,6名大便稀。在许多胃肠道症状之间发现中度相关性(r33= .396 - .751),表明这些症状可以聚类以创建一个概况。这些发现具有重要的临床意义,因为ASD的胃肠道症状由于交流和社交障碍而难以诊断,并且可能未得到诊断和治疗。由于护士经常在ASD人群的第一线进行干预,因此进一步表征该人群的胃肠道症状对护士研究人员来说是一个合乎逻辑的角色。
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引用次数: 1
I am Here. Where Are All the Other (Straight White) Male Dancers? 我在这里。其他(白人直男)男舞者在哪里?
Pub Date : 2019-05-02 DOI: 10.32473/UFJUR.V20I3.106289
Tyler Steven Renfroe
In the demographics of the American dance world—both professional and social—one group appears to be underrepresented: straight white men. Despite being the second largest group of Americans and over-represented in many other fields of endeavor, they are proportionally the least represented in social dance. Those men that do dance often began later in their lives than their female peers. Why are so many straight white men missing-in-action on the dance floor; and why, despite boundaries built by social pressure and a lack of enthusiasm or desire to change that fact, do some of “us” slip through and onto the floor? Through oral accounts, personal experience, and literature research, the research examines why so many men refuse to dance, and the motivations of those men that do dance—like me.
在美国舞蹈界的人口统计中——无论是专业的还是社会的——有一个群体似乎没有被充分代表:白人直男。尽管他们是美国人的第二大群体,在许多其他领域都有大量的代表,但在社交舞蹈中,他们的比例是最少的。那些会跳舞的男性通常比同龄的女性更晚开始跳舞。为什么有那么多白人直男在舞池里不见踪影;尽管社会压力和缺乏改变这一事实的热情或愿望建立了界限,为什么“我们”中的一些人还是会溜到地板上?通过口述、个人经历和文献研究,该研究探讨了为什么这么多男人拒绝跳舞,以及那些像我一样跳舞的男人的动机。
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引用次数: 1
UF JUR Editorial Board 2018-2019 UF JUR编辑委员会2018-2019
Pub Date : 2019-05-02 DOI: 10.32473/UFJUR.V20I3.114363
Michelle Leonard
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引用次数: 0
期刊
UF Journal of Undergraduate Research
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