Development Of Paper-Based 3d Cell Culture Devices And Real-Time Biosensing Applications,
2019
Faculty of Allied Health Sciences
Development Of Paper-Based 3d Cell Culture Devices And Real-Time Biosensing Applications, Naricha Pupinyo
Chulalongkorn University Theses and Dissertations (Chula ETD)
Due to its micro fibrous pores, paper has gained traction in the use as a scaffold for 3D cell culture. To study melanoma mechanism, screening the amount of melanin production and invasion study are major keys for drug development. In this work, we developed paper-based devices for culturing melanoma cells. Through the development process, we obtained the optimal parameters, which were applied for 3 different projects; 1) Anti-melanogenic effect screening, 2) Impedance-based E-screen assay, and 3) A membrane insert for real-time invasion assay. As a result of melanin production from melanocytes, the cells can be easily observed after a few …
Effects And Mechanisms Of Caesalpinia Mimosoides Leaf Extract On Oxidative Stress Resistance And Anti-Aging Activity In Caenorhabditis Elegans And Neurite Outgrowth Activity In Neuro2a Cells,
2019
Faculty of Allied Health Sciences
Effects And Mechanisms Of Caesalpinia Mimosoides Leaf Extract On Oxidative Stress Resistance And Anti-Aging Activity In Caenorhabditis Elegans And Neurite Outgrowth Activity In Neuro2a Cells, Panthakarn Rangsinth
Chulalongkorn University Theses and Dissertations (Chula ETD)
World’s population of older people is rising, dietary supplements promoting healthy lifespan are needed. Moreover, Alzheimer’s disease (AD), an age-related degenerative disease, becomes a public health problem in aging society. Recent reports using cell culture models of AD suggest that amyloid precursor protein (APP), a protein causally related to AD, plays an important role as an inhibitor of neurite outgrowth. Medicinal herbs with neurite outgrowth stimulatory effect may help to prevent and cure AD. Moreover, studies suggest that dietary supplements from plant sources act in preventive nutrition, since they provide antioxidant action against oxidative stress, promote healthspan and prolong lifespan. …
Development Of Cell-Penetrating Peptides Targeting Nuclear And Extra-Nuclear Signaling Pathways In Cancer Cells,
2019
Faculty of Allied Health Sciences
Development Of Cell-Penetrating Peptides Targeting Nuclear And Extra-Nuclear Signaling Pathways In Cancer Cells, Panthita Kaewjanthong
Chulalongkorn University Theses and Dissertations (Chula ETD)
Growth factors and hormones often mediate their biological effects through series of protein-protein interaction between cytoplasmic signaling molecules or transcription factors. Interfering of these protein-protein interactions has been shown to reduce or inhibit biological and growth responses to various growth factors and hormones. We previously demonstrated that progesterone receptor (PR) contains a polyproline domain (PPD) which directly interact to Src homology 3 (SH3) domain-containing molecules and expression of PR-PPD inhibits EGFR-mediated NSCLC cell proliferation. In this study, we investigated that the introduction of PR-PPD by cell-penetrating peptide could inhibit EGF-induced NSCLC cell proliferation. PR-PPD was attached to a cancer-specific CPP, …
ผลการปกป้องของสารสกัดใบรางจืดจากการเหนี่ยวนำความเป็นพิษด้วยกลูตาเมตในเซลล์ประสาท Ht-22 ผ่านวิถีออโตฟาจี,
2019
คณะสหเวชศาสตร์
ผลการปกป้องของสารสกัดใบรางจืดจากการเหนี่ยวนำความเป็นพิษด้วยกลูตาเมตในเซลล์ประสาท Ht-22 ผ่านวิถีออโตฟาจี, วุฒิพงษ์ วงศ์ทิพย์
Chulalongkorn University Theses and Dissertations (Chula ETD)
หนึ่งในสาเหตุสำคัญของการเกิดโรคความเสื่อมของระบบประสาทนั้นเกิดได้จากภาวะเครียดออกซิเดชันภายในเซลล์ประสาท โดยพบว่าสารสื่อประสาทชนิดกลูตาเมตเมื่อมีระดับสูงผิดปกติสามารถเหนี่ยวนำให้เกิดอนุมูลอิสระและภาวะเครียดออกซิเดชันนำไปสู่การสูญเสียและการตายของเซลล์ประสาทได้ โดยเฉพาะการตายของเซลล์จากการเกิดกระบวนการออโตฟาจีมากเกินไป ซึ่งเป็นกระบวนการที่มีบทบาทสำคัญในการควบคุมการตายของเซลล์ผ่านวิถีอื่นๆ รางจืด (Thunbergia laurifolia) เป็นพืชสมุนไพรไทยมีสรรพคุณในการถอนพิษ ต้านการอักเสบ และ ต้านอนุมูลอิสระได้ดี จุดประสงค์ในการวิจัยครั้งนี้ คือ ศึกษาคุณสมบัติการต้านอนุมูลอิสระของสารสกัดหยาบจากใบรางจืด และการปกป้องการตายของเซลล์ประสาทผ่านวิถีออโตฟาจี จากการเหนี่ยวนำภาวะเครียดออกซิเดชันด้วยกลูตาเมตโดยทดสอบในเซลล์ประสาทส่วนฮิปโปแคมปัสของหนูชนิด HT-22 ใบรางจืดถูกนำมาสกัดด้วยเอทานอล จากผลการทดสอบพบว่าสารสกัดหยาบจากใบรางจืดมีความสามารถในการต้านอนุมูลอิสระ และมีปริมาณสารประกอบ ฟลาโวนอยด์และฟีนอลิกสูง โดยจากการวิเคราะห์ด้วยเครื่อง LC-MS และเปรียบเทียบผลที่ได้กับฐานข้อมูล METLIN-(CA, USA) พบสารออกฤทธิ์สำคัญที่ยังไม่มีรายงานในสารสกัดเอทานอลของใบรางจืด ได้แก่ Apigenin 7-O-glucoside และ 7-Hydroxycoumarin ยิ่งไปกว่านั้นสารสกัดหยาบจากใบรางจืดยังสามารถลดการตายของเซลล์ประสาทจากการเหนี่ยวนำความเป็นพิษด้วยกลูตาเมต โดยสามารถยับยั้งการเกิดอนุมูลอิสระภายในเซลล์ และเพิ่มการแสดงออกของยีนต้านอนุมูลอิสระที่ทำหน้าที่สำคัญในกระบวนการต้านอนุมูลอิสระในร่างกาย ได้แก่ superoxide dismutase 1 และ 2 (SOD1 and SOD2) catalase (CAT) และ glutathione peroxidase (GPx) ซึ่งสัมพันธ์กับการลดการเกิดกระบวนการออโตฟาจีภายในเซลล์และเพิ่มปริมาณ ไมโตคอนเดรียโปรตีน โดยพบว่าสารสกัดหยาบจากใบรางจืดสามารถลดอัตราส่วนการแสดงออกของโปรตีน LC3B-II/LC3B-I และเพิ่มการแสดงออกของไมโตคอนเดรียโปรตีน TOM20 เมื่อทำการตรวจหาตำแหน่งที่จำเพาะต่อการแสดงออกของโปรตีนในกระบวนการออโตฟาจี (LC3B) และไมโตคอนเดรียด้วยกล้องจุลทรรศน์ชนิดคอนโฟคอล พบว่า สารสกัดหยาบจากใบรางจืดสามารถลดการเกิด LC3-puncta ซึ่งเป็นลักษณะเด่นของการเกิดกระบวนการออโตฟาจี นอกจากนี้สารสกัดหยาบจากใบรางจืดยังลดการ colocalization ของ LC3B และไมโตคอนเดรีย จากผลการศึกษานี้เราพบสารออกฤทธิ์สำคัญใหม่ที่ยังไม่มีรายงานมาก่อนของใบรางจืด โดยสารออกฤทธิ์นี้มีความสามารถในการปกป้องเซลล์ประสาทชนิด HT-22 จากภาวะเครียดออกซิเดชันและเป็นปัจจัยสำคัญทำให้สารสกัดจากใบรางจืดมีความสามารถลดการตายของเซลล์ประสาทผ่านวิถีออโตฟาจีได้ ซึ่งอาจสามารถนำไปพัฒนาเป็นสมุนไพรทางเลือกหรือยาเพื่อใช้ในการป้องกันหรือรักษาการเกิดโรคความเสื่อมของระบบประสาทได้ในอนาคต
Knob-Socket Investigation Of Stability And Specificity In Alpha-Helical Secondary And Quaternary Packing Structure,
2019
University of the Pacific
Knob-Socket Investigation Of Stability And Specificity In Alpha-Helical Secondary And Quaternary Packing Structure, Taylor Renee Rabara
University of the Pacific Theses and Dissertations
The novel knob-socket (KS) model provides a construct to interpret and analyze the direct contributions of amino acid residues to the stability in α-helical protein structures. Based on residue preferences derived from a set of protein structures, the KS construct characterizes intra- and inter-helical packing into regular patterns of simple motifs. The KS model was used in the de novo design of an α-helical homodimer, KSα1.1. Using site-directed mutagenesis, KSα1.1 point mutants were designed to selectively increase and decrease stability by relating KS propensities with changes to α-helical structure. This study suggests that the sockets from the KS Model can …
Functional Analysis Of Bacillus Anthracis Aspartate Transcarbamoylase And Dihydroorotase,
2019
Wayne State University
Functional Analysis Of Bacillus Anthracis Aspartate Transcarbamoylase And Dihydroorotase, Katelyn Leigh Schwager Silva
Wayne State University Theses
There are many enzymes required for efficient and proper pyrimidine biosynthesis. The two that are most important and were discussed in this thesis are aspartate transcarbamoylase (ATCase) and dihydroorotase (DHOase). Both play an important role in not only pyrimidine biosynthesis production, but also mechanistic regulation of de novo synthesis. Anthrax is an infection caused by Bacillus anthracis. Here we studied ATCase and DHOase in Bacillus Anthracis. In this thesis we understood the effects of the enzymes ATCase and DHOase on pyrimidine biosynthesis. Adequate inhibitors of these enzymes would result in cell death and could pose as a cure to infection …
Cyclophilin A As A Molecular Switch Regulating Prolactin Receptor Mediated Signaling, Mammary Tumorigenesis And Metastasis,
2019
Virginia Commonwealth University
Cyclophilin A As A Molecular Switch Regulating Prolactin Receptor Mediated Signaling, Mammary Tumorigenesis And Metastasis, Shawn Hakim
Theses and Dissertations
Prolactin (PRL) and its receptor (PRLr) have been implicated in the development and progression of human breast cancer. PRL activates its receptor and induces activation of proximal Janus kinase 2 (Jak2) for signal transduction. Here, we sought to determine the role of PRLr-associated peptidyl-prolyl isomerase, cyclophilin A (CypA), in modulating structure/function relationships of the PRLr. It was demonstrated that CypA mediated PRL-induced conformational change of the CFP- and -YFP tagged forms of the PRLr cytoplasmic-tail, whereas CypA inhibition by NIM811 (N-methyl-4-isoleucine cyclosporin) or knockdown blocked the conformational change of the PRLr assessed by Fluorescence Resonance Energy Transfer (FRET) signal or …
Electrostatic Networks And Mechanisms Of Δph-Dependent Gating In The Human Voltage-Gated Proton Channel Hv1,
2019
Virginia Commonwealth University
Electrostatic Networks And Mechanisms Of Δph-Dependent Gating In The Human Voltage-Gated Proton Channel Hv1, Ashley L. Bennett
Theses and Dissertations
The structure of the voltage-gated proton (H+) channel Hv1 is homologous to the voltage sensor domain (VSD) of tetrameric voltage-gated Na+, K+ and Ca2+ channels (VGCs), but lacks a pore domain and instead forms a homodimer. Similar to other VSD proteins, Hv1 is gated by changes in membrane potential (V), but unlike VGCs, voltage-dependent gating in Hv1 is modulated by changes in the transmembrane pH gradient (DpH = pHo - pHi). In Hv1, pHo or pHi changes shift the open probability (POPEN)-V relation by ~40 mV per …
Targeting Sphingosine Kinase 2 As A Treatment For Cholangiocarcinoma,
2019
Human and Molecular Genetics
Targeting Sphingosine Kinase 2 As A Treatment For Cholangiocarcinoma, Anthony D. Stillman
Theses and Dissertations
Cholangiocarcinoma (CCA) has a high mortality rate and its occurrence is rising. This increase prompts the need for improved CCA treatments. Studies have suggested that CCA is highly reliant on the sphingosine-1-phosphate-receptor-2 (S1PR2) and sphingosine kinase 2 (SphK2). Recently, a competitive SphK2 inhibitor, ABC294640, has been approved for clinical trial. ABC294640 has the potential to treat CCA, which is support by a phase I clinical study that was able to temporarily treat a patient suffering from metastasized CCA with ABC294640. To determine the viability of ABC294640 as a treatment for CCA, this study focused on determining the effects of ABC294640 …
Stat5b And Stat5 Tetramers Are Essential For Ige-Mediated Mast Cell Function,
2019
Virginia Commonwealth University
Stat5b And Stat5 Tetramers Are Essential For Ige-Mediated Mast Cell Function, Kasalina N. Kiwanuka
Theses and Dissertations
Signal Transducers and Activators of Transcription (STATs) are latent transcription factors that mediate several cellular responses. This protein family consists of seven members, STAT1 – 6 including two closely related molecules, STAT5a and STAT5b, that show 96% amino acid sequence homology and are critical for lymphoid, myeloid and erythroid cell development and function. Activated STAT proteins dimerize and translocate to the nucleus, where they bind to high-affinity DNA motifs to modulate gene expression. We recently identified STAT5b as the critical regulator of IgE-mediated cytokine production in mast cells. STAT5b knockout (KO) cells show decreased sensitivity to IgE-mediated passive systemic anaphylaxis …
Expression, Purification, Functional Characterization And Crystallization Of Three Porphyromonas Gingivalis Gene Products,
2019
Wilfrid Laurier University
Expression, Purification, Functional Characterization And Crystallization Of Three Porphyromonas Gingivalis Gene Products, Katarina Mandic
Theses and Dissertations (Comprehensive)
Dental biofilms are coupled polymicrobial aggregates that have attached to solid surfaces in the oral cavity. These collections of microorganisms are known to cause periodontal diseases that commence as localized inflammation of the gingiva and if untreated, eventually lead to irreversible alveolar bone resorption and tooth loss. Porphyromonas gingivalis is one of three periodontal pathogens that make up the “Red Complex”; a bacterial consortium responsible for the production of polysaccharide-rich biofilms that are essential to the inception and progression of periodontal disease. The dysbiosis and destructive inflammation caused by these organisms propel a self-sustained feed-forward loop that perpetuates periodontal disease. …
Structural And Functional Analysis Of Three Upregulated Gene Products, Tde0626, Tde1701, And Tde2714 From Treponema Denticola During Biofilm Formation,
2019
Wilfrid Laurier University
Structural And Functional Analysis Of Three Upregulated Gene Products, Tde0626, Tde1701, And Tde2714 From Treponema Denticola During Biofilm Formation, Jonah Nechacov
Theses and Dissertations (Comprehensive)
The progression of human chronic periodontitis within periodontal disease has been often linked to the presence of key pathogens, such as the presence of Treponema denticola, a late colonizer found in the deepening pockets of the gingival sulcus. This pathogen, as well as its associates Porphyromonas gingivalis and Tannerella forsythia, are classified as the ‘red complex’ and exist in a mixed biofilm during infection. It is within this biofilm state that previous transcriptomic analysis revealed a total of 126 genes that had an increase in their expression by 1.5-fold or greater in T. denticola. Three of these …
Lost In Translation: Challenges And Solutions To Language Barriers In Healthcare In Mississippi,
2019
University of Mississippi
Lost In Translation: Challenges And Solutions To Language Barriers In Healthcare In Mississippi, Galina Ostrovsky
Honors Theses
Language barriers can impede access to healthcare and compromise quality of care for limited English proficiency (LEP) patients. Although a rights-based framework would suggest and federal mandates require healthcare facilities to provide language access services, multiple factors result in the persistence of language barriers in healthcare. As the literature review shows, these factors include limitations in policy, enforcement, standardization, resources, and ability to recruit and retain professional interpreters. This thesis uses a multi-method case study entailing Mississippi in general and information from two specific sites, a rural Delta county and an urban central county, to assess the issue of language …
Characterization Of The Red Complex Bacterial Biofilm,
2019
Wilfrid Laurier University
Characterization Of The Red Complex Bacterial Biofilm, Alena Pratasouskaya
Theses and Dissertations (Comprehensive)
Commonly associated with severe inflammation and destruction of the tooth-supporting tissue is bacterial consortia Treponema denticola, Tannerella forsythia, and Porphyromonas gingivalis, collectively referred to as the red complex. The red complex uses the common bacterial strategy of producing and embedding itself in extracellular polymeric substance, which contributes to the recalcitrance of periodontitis and was therefore of interest in this study. The red complex static cultures were grown in combination with different chemicals in order to establish what changes accompany these chemical challenges. This research established that extracellular polymeric substance carbohydrate yields increased with time. Interestingly, carbohydrate composition …
Characterization Of Wssf; A Putative Acetyltransferase From Achromobacter Insuavis And Pseudomonas Fluorescens,
2019
Wilfrid Laurier University
Characterization Of Wssf; A Putative Acetyltransferase From Achromobacter Insuavis And Pseudomonas Fluorescens, Cody Reese
Theses and Dissertations (Comprehensive)
Biofilms are a survival mechanism commonly employed by communities of bacteria for adherence and protection. Bacteria produce a matrix of polymers (e.g. exopolysaccharides, such as cellulose) that allow them to exert control on their local environment. In the case of cellulose biofilms, acetylation (addition of acetate on carbohydrates) is paramount for polymer integrity and in some cases virulence. For this research, the wrinkly spreader (WS) genotype of the emergent human pathogen Achromobacter insuavis facilitates infections of the eyes of contact lens wearers and the lungs of Cystic Fibrosis patients (CF). Chronic infections have created a growing concern for the protective …
Functional Characterization Of O-Acetyltransferase Wssi: Role In Bacterial Cellulose Acetylation Of Achromobacter Insuavis And Pseudomonas Fluorescens,
2019
Wilfrid Laurier University
Functional Characterization Of O-Acetyltransferase Wssi: Role In Bacterial Cellulose Acetylation Of Achromobacter Insuavis And Pseudomonas Fluorescens, Alysha Burnett
Theses and Dissertations (Comprehensive)
Cellulose biofilms consist of a community of bacterial cells encased in a self-produced matrix of polymers (e.g. exopolysaccharides, such as cellulose) that facilitate a firm adherence to surfaces. The acetylation (addition of an acetyl group on carbohydrates) is crucial in virulence; thereby in some cases allowing opportunistic bacteria to cause harmful diseases. Pertaining to this research, the wrinkly spreader (WS) genotype of Pseudomonas fluorescens colonizes the air-liquid interface on food, water sources and human tissue to form a robust biofilm with the ability to spread across surfaces. The composition of this biofilm largely consists of bacterial cellulose polymers …
Structural Investigation Of Bcsc: Insight Into Periplasmic Transport During Cellulose Export,
2019
Wilfrid Laurier University
Structural Investigation Of Bcsc: Insight Into Periplasmic Transport During Cellulose Export, William Scott, Joel T. Weadge
Theses and Dissertations (Comprehensive)
A biofilm can be defined by a community of microbes coexisting within a self-produced protective polymeric matrix. Exopolysaccharide (EPS) is a key component in biofilms and a contributor to their virulence and pathogenicity. The cellulose bacterial synthesis complex is one such EPS system that is found in many Enterobacteriaceae,including Escherichia coli and Salmonella spp., and is responsible for the production and secretion of the EPS cellulose. BcsC is the periplasmic protein responsible for the export of the exopolysaccharide cellulose and was the focus of this research. Sequence homology comparisons and structural predictions between BcsC, and the previously characterized alginate …
Near-Infrared Fluorescent Probes For Sensitive Determination Of Lysosomal & Mitochondrial Ph In Live Cells,
2019
Michigan Technological University
Near-Infrared Fluorescent Probes For Sensitive Determination Of Lysosomal & Mitochondrial Ph In Live Cells, Wafa Mazi
Dissertations, Master's Theses and Master's Reports
Varied intracellular pH levels are critical for various physiological processes such as enzymatic activity, cell proliferation and apoptosis, ion transport, and muscle contraction. Cellular compartments, like lysosomes, must retain an acidic environment (pH ~ 4.5) to activate hydrolytic enzymes necessary for the breakdown of large biomolecules. Another cellular organelle, the mitochondria, provides the cell with energy and must retain an alkalis environment (pH ~ 8.0) for proper function. Substantial lysosomal and mitochondrial pH deviation is associated with cellular dysfunction and disease. Therefore, the precise detection of lysosomal and mitochondrial pH is essential to provide a better understanding of cellular physiological …
Characterization Of The Microbial Phosphonate-Activating Pntc Enzymes,
2019
Wilfrid Laurier University
Characterization Of The Microbial Phosphonate-Activating Pntc Enzymes, Kyle Rice
Theses and Dissertations (Comprehensive)
New strategies are urgently needed to combat infectious diseases in an era of rising antibiotic resistance. Furthermore, an emerging appreciation for the human microbiome’s role in maintaining health motivates discovery of species-specific antibiotics that minimally disrupt our native bacterial communities. Small molecule modifications to bacterial cell surfaces represent a potentially rich source of new targets for next generation antibiotics, as these molecules mediate virulence and evasion of the host immune response. Phosphocholine (PCho) is a rare cell surface modification that contributes to virulence, and modifications with phosphonates like 2-aminoethylphosphonate (AEP) are even more unusual and therefore provide opportunities for species- …
In Silico Development Of An Rna Aptamer Library To Be Use For The Selection Of Rna Aptamer That Target Biomolecules,
2018
Pittsburg State University
In Silico Development Of An Rna Aptamer Library To Be Use For The Selection Of Rna Aptamer That Target Biomolecules, Nehad Nawfawi
Electronic Theses & Dissertations
The systematic evolution of ligands by exponential enrichment (SELEX) is a powerful method for the development of high affinity RNA ligands toward and infinite array of target molecules. SELEX is based upon the generation of a randomized population of RNA or DNA molecules followed by a target molecule that selects high affinity ligands from the randomized population followed by the subsequent amplification of the selected molecules. The procedure of selection and amplification is typically carried out through multiple cycles to insure that the identified ligands exhibits the highest affinity toward the target. The procedure is very time- consuming often taking …
