Biochemistry, Biophysics, and Structural Biology Commons^™

Characterization Of A Malachite Green Dna Aptamer And Development Of Malachite Green Dna Aptamer Sensor, Martin O'Steen

Electronic Theses and Dissertations, 2020-2023

Hydrogels have been extensively studied for use in applications such as tissue engineering, drug delivery systems, wound dressing, actuators, valves, and sensors in Micromechanical Systems (MEMS), among others. Thermo-responsive hydrogels in particular pose various advantages such as their capability to respond to an external stimulus and reverse the response when the stimulus is removed and the ability to imbibe a significant amount of solvent and increase their volume by over 1000%. Extensive research has been conducted to tune and improve hydrogel's rate and degree of swelling and mechanical properties. Previous work showed that when synthetic clay (Laponite) is cross-linked with …

Fish Oil Intake During Gestation And Lactation Attenuated Stz-Induced Diabetes Inmale Offspring Via Activation Of Brown Fat And Modulating Oxylipin Profile, Rong Fan, Shalom Thomas, Mikyoung You, Zhuoheng Li, Brandt Bessell, Bhanwar Lal Puniya, Tomáš Helikar, Zhenhua Liu, Soonkyu Chung

Department of Biochemistry: Faculty Publications

Fish oil (FO) has been demonstrated to activate brown thermogenesis and attenuate inflammation in the brown adipose tissue (BAT). Previously, we have reported thatmaternal FO supplementation promotes BAT activity of the weaned mice pups. However, whether maternal FO intake could confer sustainable metabolic benefits to offspring remains uncovered. Therefore, this study aimed to determine the differential impact of maternal FO during pregnancy versus lactation on BAT transcriptome and evaluate the role of bioactive lipid metabolites derived from maternal FO supplementation on the extended metabolic benefits of older pups in the context of type 1 diabetes (T1D).

Conclusions: Our results suggested …

Recognition Of Extracellular Dna By Type Iv Pili Promotes Biofilm Formation By Clostridioides Difficile, Leslie A. Ronish, Ben Sidner, Yafan Yu, Kurt H. Piepenbrink

Department of Biochemistry: Faculty Publications

Clostridioides difficile is a Gram-positive bacillus, which is a frequent cause of gastrointestinal infections triggered by the depletion of the gut microbiome. Because of the frequent recurrence of these infections after antibiotic treatment, mechanisms of C. difficile persistence and recurrence, including biofilm formation, are of increasing interest. Previously, our group and others found that type IV pili, filamentous helical appendages polymerized from protein subunits, promoted microcolony and biofilm formation in C. difficile. In Gram-negative bacteria, the ability of type IV pili to mediate bacterial self-association has been explained through interactions between the pili of adjacent cells, but type …

Coordination Of Metal Center Biogenesis In Human Cytochrome C Oxidase, Eva Nývltová, Jonathan Dietz, Javier Seravalli, Oleh Khalimonchuk, Antoni Barrientos

Department of Biochemistry: Faculty Publications

Mitochondrial cytochrome c oxidase (CcO) or respiratory chain complex IV is a heme aa3- copper oxygen reductase containing metal centers essential for holo-complex biogenesis and enzymatic function that are assembled by subunit-specific metallochaperones. The enzyme has two copper sites located in the catalytic core subunits. The COX1 subunit harbors the CuB site that tightly associates with heme a3 while the COX2 subunit contains the binuclear CuA site. Here, we report that in human cells the CcO copper chaperones form macromolecular assemblies and cooperate with several twin CX9C proteins to control heme a biosynthesis and coordinate copper transfer sequentially to the …

Anti-Neuroinflammatory Effects And Underlying Mechanismsof Cleistocalyx Nervosum Var. Paniala Extracts In Microglia Cells, Sakawrat Janpaijit

Chulalongkorn University Theses and Dissertations (Chula ETD)

Sustained inflammatory responses have been implicated in various neurodegenerative disorders. Cleistocalyx nervosum var. paniala (CN), an edible berry, has been reported to exhibit several beneficial properties. However, only limited research is available on this fruit seed, which is classified as agricultural food waste. We therefore focused on the anti-neuroinflammatory effects and underlying mechanism of C. nervosum var. paniala seed extract (CNSE) on BV-2 mouse microglial cells induced by lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-α). CN seeds were extracted in 95% ethanol by Soxhlet extraction. HPLC was carried out to identified resveratrol (RESV) in CNSE. For cell-based studies, the expression …

Kinetic Mechanism Of Indigo-Producing Indole Monooxygenase From Acinetobacter Baumannii, Kanyarat Suksomjaisaman

Chulalongkorn University Theses and Dissertations (Chula ETD)

Indole, a bicyclic compound comprising a fused benzene and pyrrole ring, serves as a fundamental component in numerous natural compounds. Additionally, indole holds significant importance in industrial applications, particularly in the production of pharmaceutical drugs and agrochemicals. Flavin-dependent indole monooxygenase is an enzyme responsible for hydroxylation of the indole ring. Indole monooxygenase is a two-components: flavoprotein reductase containing FAD as a cofactor, and monooxygenase. The reductase component utilizes NADH to catalyze flavin reduction and transfers the reduced flavin to the monooxygenase component. The hydroxylation of indole by monooxygenase reduced FAD and oxygen as substrates. In this study, we cloned the …

Microbial Labilization And Diversification Of Pyrogenic Dissolved Organic Matter, Aleksandar I. Goranov, Andrew S. Wozniak, Kyle W. Bostick, Andrew R. Zimmerman, Siddhartha Mitra, Patrick G. Hatcher

Chemistry & Biochemistry Faculty Publications

With the increased occurrence of wildfires around the world, interest in the chemistry of pyrogenic organic matter (pyOM) and its fate in the environment has increased. Upon leaching from soils by rain events, significant amounts of dissolved pyOM (pyDOM) enter the aquatic environment and interact with microbial communities that are essential for cycling organic matter within the different biogeochemical cycles. To evaluate the biodegradability of pyDOM, aqueous extracts of laboratory-produced biochars were incubated with soil microbes, and the molecular changes to the composition of pyDOM were probed using ultrahigh-resolution mass spectrometry (Fourier transform–ion cyclotron resonance–mass spectrometry). Given that solar irradiation …

The Role Of Irf-1 In Spontaneous Mouse Glioma, Aakash B. Vaidya

Theses and Dissertations

Glioblastoma Multiforme has been shown to be one of the deadliest primary brain cancers. One of the reasons why GBM is so deadly, is a unique immunosuppressive tumor microenvironment that promotes GBM growth and progression. Both astrocyte and microglia have been implicated in immunosuppression. In this study, we explored the role of Interferon Regulatory Factor 1 (IRF-1) in astrocytes and glioma cells on the growth of spontaneous glioma tumors. IRF-1 is regulated by the JAK/STAT pathway and induces expression of Programmed death ligand 1 (PD-L1). PD-L1 downregulates immune responses to glioma. We found that IRF-1 had no effect on spontaneous …

Targeting Bcl-2 Family Proteins In Therapy Induced Senescent Cancer Cell Models, Wade Cook

Theses and Dissertations

Non-small Cell Lung Cancer (NSCLC) originates from numerous different cell types in the lungs and is among the deadliest of cancers. Head and Neck Squamous Cell Carcinomas (HNSCC) are derived from the mucosal membranes of the oral cavity, pharynx, and larynx. Both NSCLC and HNSCC are predominately caused by tobacco smoke inhalation and as such mutations in the tumor suppressor gene TP53 are common. Since similarities exist in the root cause of NSCLC and HNSCC, they may also share similarities in treatment methods. Cisplatin is a platinum-based DNA damaging agent that has been used as a cancer chemotherapy for decades. …

Structure Of A Monomeric Photosystem Ii Core Complex From A Cyanobacterium Acclimated To Far-Red Light Reveals The Functions Of Chlorophylls D And F, Christopher J. Gisriel, Gaozhong Shen, Ming-Yang Ho, Vasily Kurashov, David A. Flesher

Faculty Publications

Far-red light (FRL) photoacclimation in cyanobacteria provides a selective growth advantage for some terrestrial cyanobacteria by expanding the range of photosynthetically active radiation to include far-red/near-infrared light (700-800 nm). During this photoacclimation process, photosystem II (PSII), the water:plastoquinone photooxidoreductase involved in oxygenic photosynthesis, is modified. The resulting FRL-PSII is comprised of FRL-specific core subunits and binds chlorophyll (Chl) d and Chl f molecules in place of several of the Chl a molecules found when cells are grown in visible light. These new Chls effectively lower the energy canonically thought to define the "red limit" for light required to drive photochemical …

Leveraging Queueing Theory To Develop Advanced Synthetic Biological Circuits, Prajakta K. Jadhav

Electronic Theses and Dissertations

The complexity and redundancy in the network topology of the cell hinder our understanding of the natural system, making it challenging to engineer living organisms with the desired functionality. The long-term goal of synthetic biology and bioengineering is to engineer cells to perform specific functions with increased robustness. The robust engineered systems can be easily applied to different species with the same outcome. An effective approach is a bottom-up approach where we tease apart the biological pathways and study them independently. Understanding the underlying mechanisms of biological pathways can be further applied to construct complex biological circuits that function similarly …

Understanding The Impact Of Human Germline Single-Nucleotide Variants, Samuel Adjei

Electronic Theses and Dissertations

All genetic variations in the human genome are due to mutations, and therefore determining the impact of the different categories of mutations, particularly nonsynonymous single nucleotide variants (SNVs) which are the most abundant in the population is essential for understanding the genetics of human diseases. Generations of mutation studies have focused on a relatively small number, partly due to technological limitations. However, advances in next-generation sequencing have allowed for empirical assessments of genome-wide mutations. genome sequence of any individual contains extensive protein-altering genetic variation (missense mutations) and of which only a few are unambiguously deleterious. Characterizing the phenotypes of these …

Increased Cd11b+ Immune Cells Contribute To Thoracic Aortic Aneurysm Rupture, Lucynda Pham

Wayne State University Theses

A life-threatening clinical manifestation of Marfan Syndrome (MFS) is the development of thoracic aortic aneurysms (TAA) which can lead to dissection and or rupture. MFS is caused by mutations in extracellular matrix (ECM) protein, Fibrillin-1 (FBN1) which forms the structural component of elastic microfibrils. It is unclear how mutations in FBN1 cause aneurysm development and progression. In recent years, inflammation and immune cells are being considered an important biological process that contributes to TAA. However, the exact roles of the immune cells are unknown. Here we find increased CD11b+ cells recruited and infiltrated into the vessel wall and their spatial …

Growing Meat On Plants: Using Intermediate Cbd-Rgd Fusion Proteins To Improve Bovine Satellite Cell Attachment On Cellulose-Based Scaffolds, Julian Cohen

Pitzer Senior Theses

Cellular agriculture is an emerging technology aiming to replace existing methods for animal agriculture with tissue engineering and cell culture-based technologies. Cultured meat falls within this purview, using a biomimetic approach to recreate animal muscle tissue through tissue engineering. In the attempt to diminish the necessity of animal-derived materials within this process, plant-based scaffolds can be used as a substrate upon which stem cells are cultured. Due to the unfavorable environment of cellulose for mammalian cell-surface proteins, the approach was taken of coating cellulose nanofiber films with a fusion protein composed of a cellulose binding domain (CBD) protein and the …

Characterization Of L-Malic Acid-Capped Cds Quantum Dots And Examination Of The Nanoncrystal's Biosensor Ability, Andrei A. Potapenko

Honors Theses and Capstones

Ligand-capped quantum dots are microscopic semiconductors commonly used in biomedical research and therapy for cell imaging and drug delivery. Despite the abundance of in vivo applications, the nature of interactions between the nanocrystals and the amino acids comprising human proteins remains insufficiently investigated. In this work, we report on the synthesis, ligand exchange, and characterization of the L-malic acid-capped CdS quantum dots via various spectroscopic techniques. We also assess the ligand-capped nanocrystals as potential probes for some amino acids. We determine that in the presence of L-cysteine, the L-malic acid-capped CdS quantum dots fluorescence signal quenches drastically, thus indicating a …

Inhibition Of Lysine Acetyltransferases Kat 3a/3b And Its Effect On Poliovirus Proliferation, Eduards Norkvests

Legacy Theses & Dissertations (2009 - 2024)

Ribosomes have been traditionally viewed as invariable machinery, but in recent years more and more evidence for their heterogeneity has emerged. Core ribosomal proteins, ribosomal-associated proteins, and post-translational modifications all contribute to the variations found within these crucial subcellular components.In our research, we focused on lysine acetylation – a post translational modification found on many ribosomal proteins. We used a chemical inhibitor of acetyltransferases KAT 3A and KAT 3B to observe the effect that inhibiting acetylation may have in the context of translation. By using poliovirus to shut off conventional cellular translation, we were able to look at a narrow …

Salivary Gland Stromal Heterogeneity And Epithelial Controls, Nicholas L. Moskwa

Legacy Theses & Dissertations (2009 - 2024)

Organogenesis is the process organs go through where cellular communications coordinate all a developing organ needs. What organs need are more cells, in the right place, doing the right job. In the salivary gland, we know that stromal cells are important for organogenesis and that they coordinate the epithelium’s form and functions. However, specific stromal contributions have focused on epithelial quantity and placement. There is less information about how the stroma directs the epithelium towards certain functions. Here we used organoids as a model for understanding what stromal signaling directs epithelial cell fate. We found that stromal cell state is …

Synthesis Of Oligonucleotides Containing Unnatural Backbone And Regulation Of Crispr Activity Using Inverse Electron Demand Diels-Alder Chemistry, Alyssa Hoy

Legacy Theses & Dissertations (2009 - 2024)

This thesis describes the synthesis of an oligonucleotide containing an unnatural neutralbackbone. The backbone was constructed using bio-orthogonal inverse electron demand Diels- Alder chemistry between trans-cyclooctene (TCO) and tetrazine (Tz). A TCO phosphoramidite was synthesized and attached to solid support for strand propagation. Uridine monomers containing either bis-TCO or bis-Tz were also synthesized. The oligonucleotide strand was formed by sequential ligation of the bis-Tz and bis-TCO monomers. The oligonucleotide was analyzed using mass spectrometry. This thesis also describes the synthesis of sgRNAs containing non-canonical nucleobases, m1A, m6A, s2U and s4U using the ‘split-and-click’ methodology. The synthetic sgRNAs were assembled using …

Exploring The Roles Of Stress, Codon Usage, And Rna Modifications In Myotonic Dystrophy Type 1, Afrooz Golestanian

Legacy Theses & Dissertations (2009 - 2024)

Myotonic dystrophy (DM), the most common form of muscular dystrophy, is a neuromuscular disease caused by microsatellite repeat expansions. It can represent a multi-systemic autosomal dominant disease with DM1 and DM2 subtypes. A cytosine-thymine-guanine (CTG) triplet repeat in the 3’ untranslated region (3’UTR) of myotonic dystrophy protein kinase (DMPK) gene causes DM1 disease, which leads to the production of a longer, abnormal and toxic mRNA. The toxic DMPK mRNA sequester the splicing proteins such as Muscle blind-like (MBNL) and rbFOX which leads to gene expression alteration. Repeat associated non-AUG (RAN) translation also occurs in DM1. Mitochondrial dysregulation has also been …

The Photo-Transformation Of Free Methionine In The Presence Of Surrogate And Standard Isolate Dissolved Organic Matter Under Sunlit Irradiation, Benjamin J. Mohrhardt

Dissertations, Master's Theses and Master's Reports

Sulfur (S)-containing amino acids are key sources of carbon, nitrogen, and sulfur involved in protein synthesis, protein function, and providing energy for microbial growth. Dissolved free and combined methionine is one of two S-containing amino acids incorporated into proteins and has been attributed to their stability and function. The oxidation of methionine has received considerable attention given its ubiquitous presence in most biological systems and has been associated with losses in protein function and pathological disorders. In natural waters, methionine is rapidly and selectively taken up by microorganisms to achieve cellular requirements of carbon, nitrogen, and sulfur. The abiotic transformation …

Dissecting The Cellular Landscape And Transcriptome Network In Viral Myocarditis By Single-Cell Rna Sequencing, Ninaad Lasrado, Nicholas Borcherding, Rajkumar Arumugam, Timothy K. Starr, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Coxsackievirus B3 (CVB3)-induced myocarditis is commonly employed to study viral pathogenesis in mice.Chronically affected micemaydevelop dilated cardiomyopathy, which may involve the mediation of immune and nonimmune cells. To dissect this complexity, we performed single-cell RNA sequencing on heart cells from healthy and myocarditic mice, leading us to note significant proportions of myeloid cells, T cells, and fibroblasts. Although the transcriptomes of myeloid cells were mainly of M2 phenotype, the Th17 cells, CTLs, and Treg cells had signatures critical for cytotoxic functions. Fibroblasts were heterogeneous expressing genes important in fibrosis and regulation of inflammation and immune responses. The intercellular communication networks …

Identification And Characterization Of Transcription Factors Regulating Methionine Gamma-Lyase Gene Expressionin Durian Durio Zibethinus M. Pulps, Pinnapat Pinsorn

Chulalongkorn University Theses and Dissertations (Chula ETD)

Durian (Durio zibethinus L.) is an economic fruit crop in Southeast Asia. In Thailand, more than 200 durian varieties are found. Among them, two commercial cultivars which are Chanee and Monthong are mainly cultivated. Durian fruit releases a strong sulfuryl odor containing ethanethiol, methanethiol, and hydrogen sulfide during ripening. Methionine γ-lyase (MGL) is an enzyme responsible for α, γ-elimination of L-methionine into methanethiol, ammonia, and α-ketobutyrate. MGL gene is highly expressed in durian pulps during ripening concurring with methanethiol production during ripening. However, the function of durian MGL (DzMGL) has not been characterized. Thus, the enzymatic activity assay towards L-methionine, …

Challenges Of Constructing Entrainment Map For Arbitrary Circadian Models, Yuxuan (Nelson) Wu

Honors Theses

The entrainment map, developed by Dr.Diekman and Dr.Bose, is claimed to be a 1-dimensional map that produces a better prediction for phase-locking than methods than the phase response curve for circadian models. In his paper, he constructs the entrainment map for the two-dimensional circadian model, the Novak-Tyson model, and the other two higher-dimensional circadian models. For this thesis, we concentrate on exploring if it is viable to construct the entrainment map for other circadian models that are not included in his paper: the Becker-Weimann model and the Relogio model. In addition, we discuss the challenges of constructing the entrainment map …

Identification Of Glutathionylated Cysteines Regulating Cell Migration, Dhanushika Kukulage

Wayne State University Dissertations

Cell migration is vital in many physiological processes, such as wound repair and metastasis. Impaired cell migration in wound healing may result in chronic wounds, by which 6.7 million Americans are affected, and they suffer from bacterial infections, amputations, increased mortality, and depression. Reactive oxygen species (ROS) or hydrogen peroxide (H2O2) play an essential role as a second messenger in redox signaling. Evidence has been found that H2O2 takes part in regulating cell migration as well. A primary mechanism of ROS regulating redox signaling is oxidative protein modifications. Protein S-glutathionylation, one of the oxidative modifications, is the conjugation of glutathione …

Genetic Analysis Of Vibrio Cholerae During Its Hyperinfectious State In Danio Rerio (Zebrafish), Ravipaul Singh

Wayne State University Theses

ABSTRACTGENETIC ANALYSIS OF VIBRIO CHOLERAE DURING ITS HYPERINFECTIOUS STATE IN DANIO RERIO (ZEBRAFISH) by RAVIPAUL SINGH AUGUST 2022 Advisor: Dr. Jeffrey Withey Major: Biochemistry and Molecular Biology Degree: Master of Science Vibrio cholerae is a gram-negative, comma-shaped, monotrichous bacillus and the causative agent of the diarrheal disease cholera. V. cholerae is a natural resident of aquatic environments such as rivers and estuaries and is a highly motile bacterium. Upon entering the human host, V. cholerae travels to its site of colonization in the small intestine and upregulates its virulence factors. Upon escape from the host, V. cholerae can enter a …

Effect Of Allosteric Site Binding On Smyd2 Conformational Ensemble, Jacob Daniel Sobota

Wayne State University Theses

Molecular dynamics simulation allows for the simulation of proteins with precise control over what ligands are present at what sites. The conformational ensemble of a protein can be obtained from MD simulation with and without binding at an allosteric site and compared between to gain insight at what effects binding at an allosteric site has on a protein’s structure. SMYD2, a critical regulator of cellular processes, was recently found to have a novel secondary binding site with allosteric effects on its activity. The discovery of an allosteric site has implications for drug design in the inhibition of SMYD2 to treat …

Mhc Diversity Analysis Of Spheniscus Demersus For In Situ And Ex Situ Populations, Antonieta Van Den Berg Monsalve

Undergraduate Honors Thesis Projects

The South African Penguin has been listed as endangered by the International Union for Conservation of Nature. Major histocompatibility complex (MHC) alleles provide valuable statistics because of their variation in terms of vertebrate genomes and are pertinent to immune and reproductive health. A greater number of MHC alleles correlates with survivability of a population because the genes control the immune and reproductive systems. Legacy research by Otterbein alumni and Dr. Simon Lawrance studied MHC samples from wild, in situ, and captive, ex situ, penguin populations. By comparing these populations' major histocompatibility complexes through biostatistical analysis, contributions to conservation of the …

College Of Natural Sciences Newsletter, January 2022, College Of Natural Sciences

College of Natural Sciences Newsletters and Reports

Volume 3, Issue 1

Page 1 Dean's Message
Page 2 Awards & Recognition
Page 3 Outreach in Harrisburg, SD
Page 4 Geography Alumni Named Among 'Highly Cited Researchers', 53rd Annual Geography Convention
Page 5 Geography Convention Speakers, Brookings/SDSU Day
Page 6 Media Coverage of Natural Sciences
Page 7 Open PRAIRIE Data
Page 8 Fall 2021 Dean's List
Page 9 Fall 2021 Dean's List, cont.

ฮอร์โมนเอสโตรเจนเหนี่ยวนำให้เกิดการแทรกของ Line-1 ดีเอ็นเอเข้าไปในจีโนมของเซลล์มะเร็งเต้านม, กุลจิรา โรจน์ไพศาลกิจ

Chulalongkorn University Theses and Dissertations (Chula ETD)

ในปัจจุบันโรคมะเร็งเต้านมเป็นมะเร็งที่มีอุบัติการณ์และอัตราการตายสูงที่สุดในเพศหญิงทั่วโลกรวมถึงในประเทศไทยด้วย โดยพบว่า LINE-1 ถือว่าเป็น Retrotransposon element ที่พบมากที่สุดถึง 17 % ของจีโนมและมีความสามารถในการคัดลอกแล้วแทรกตัวเองในบริเวณอื่นของจีโนมซึ่งเป็นสาเหตุของทำให้เกิดความไม่สเถียรของจีโนมอันเป็นสาเหตุของการเกิดโรคมะเร็ง และยังพบว่าการแสดงออกของLINE-1 มีผลต่อการลุกลาม, ขนาด, และระยะของโรคมะเร็งเต้านมด้วย นอกจากนี้ยังพบว่าผู้หญิงที่การได้รับฮอร์โมนเอสโตรเจนต่อเนื่องเป็นระยะเวลานานทำให้ความเสี่ยงของโรคมะเร็งเต้านมเพิ่มมากขึ้น ถึงกระนั้นยังคงไม่ทราบถึงกลไกการทำงานของเอสโตรเจน ดังนั้นผู้วิจัยจึงตั้งสมมุติฐานว่า เอสโตรเจนสามารถเหนี่ยวนำให้เกิดการแทรกของ LINE-1 ดีเอ็นเอเข้าไปในจีโนมของเซลล์มะเร็งเต้านม โดยทำการศึกษาบผลของเอสโตรเจนที่ได้รับต่อการแสดงออกของ LINE-1 mRNA และ LINE-1 โปรตีน รวมถึงผลของเอสโตรเจนที่ได้รับต่อการแทรกของ LINE-1 ดีเอ็นเอโดยวิเคราะห์จากปริมาณดีเอ็นเอในเซลล์มะเร็งเต้านม T47D ผลการทดลองพบว่าที่เอสโตรเจนมีผลทำให้การแสดงออกของ LINE-1 mRNA เพิ่มขึ้นสูงที่สุดอย่างมีนัยสำคัญที่ความเข้มข้นของเอสโตรเจน 5 nM ด้วยวิธี Realtime RT-PCR ในเซลล์มะเร็งเต้านม T47D และผลทำให้การแสดงออกของ LINE-1 mRNA เพิ่มขึ้นสูงที่สุดอย่างมีนัยสำคัญที่ 12 ชม. ด้วยวิธี Realtime RT-PCR ในเซลล์มะเร็งเต้านม T47D และเอสโตรเจนมีผลทำให้การแสดงออกของโปรตีน LINE-1 ORF1p เพิ่มขึ้นสูงที่สุดอย่างมีนัยสำคัญที่ 48 และ 72 ชม. ในเซลล์มะเร็งเต้านม T47D ด้วยวิธี Western blot โดยพบว่าการแสดงออกของโปรตีน LINE-1 ORF1p เพิ่มขึ้นสูงที่สุดอย่างมีนัยสำคัญ 72 ชม. นอกจากนี้เอสโตรเจนมีผลทำให้ปริมาณของ LINE-1 ดีเอ็นเอบริเวณ ORF2 เพิ่มขึ้นสูงที่สุดอย่างมีนัยสำคัญที่ 72 ชม.ในเซลล์มะเร็งเต้านม T47D มากที่สุดอย่างมีนัยสำคัญด้วยวิธี Real-time PCR แสดงให้เห็นว่าเอสโตรเจนมีผลทำให้เกิดการแทรกของ LINE-1 ดีเอ็นเอเข้าไปในจีโนมได้ จากผลการทดลองทั้งหมดจึงสรุปได้ว่า เอสโตรเจนมีผลต่อการเพิ่มแสดงออกของ LINE-1 mRNA และ LINE-1 โปรตีน รวมถึงเพิ่มปริมาณ LINE-1 ดีเอ็นเอ อย่างมีนัยสำคัญในเซลล์มะเร็งเต้านม T47D แสดงว่าเอสโตรเจนสามารถเหนี่ยวนำให้เกิดการแทรกของ LINE-1 ดีเอ็นเอเข้าไปในจีโนมนำไปสู่การกลายพันธุ์ของยีนและความไม่เสถียรของจีโนม

Molecular Mechanism Of Damage-Associated Molecular Patterns In Association With Acute Hepatopancreatic Necrosis Disease Resistance Of Heat Stressed-Shrimp Penaeus Vannamei, Supitcha Wanvimonsuk

Chulalongkorn University Theses and Dissertations (Chula ETD)

Non-lethal heat shock (NLHS) is known to enhance resistance to acute hepatopancreatic necrosis disease caused by Vibrio parahaemolyticus (VPAHPND) in the Pacific white shrimp Penaeus vannamei. Generally, cell damage induced by stresses leads to the release of damage-associated molecular patterns (DAMPs), molecules that act as endogenous danger signals to promote immune response. The first part of this research aims to study molecular mechanism of DAMPs that can enhance innate immunity upon NLHS treatment on VPAHPND resistance. We identified and characterized the candidate NLHS-induced DAMP that can activate innate immune responses against VPAHPND. The candidate DAMP genes were retrieved from the …