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Nanosecond Pulsed Electric Field Induced Changes In Cell Surface Charge Density, Diganta Dutta, Xavier-Lewis Palmer, Anthony Asmar, Michael Stacey, Shizhi Qian 2017 Old Dominion University

Nanosecond Pulsed Electric Field Induced Changes In Cell Surface Charge Density, Diganta Dutta, Xavier-Lewis Palmer, Anthony Asmar, Michael Stacey, Shizhi Qian

Bioelectrics Publications

This study reports that the surface charge density changes in Jurkat cells with the application of single 60 nanosecond pulse electric fields, using atomic force microscopy. Using an atomic force microscope tip and Jurkat cells on silica in a 0.01 M KCl ionic concentration, we were able to measure the interfacial forces, while also predicting surface charge densities of both Jurkat cell and silica surfaces. The most important finding is that the pulsing conditions varyingly reduced the cells' surface charge density. This offers a novel way in which to examine cellular effects of pulsed electric fields that may lead to …


Structural And Mechanistic Insights Into Hemoglobincatalyzed Hydrogen Sulfide Oxidation And The Fate Of Polysulfide Products, Victor Vitvitsky, Pramod K. Yadav, Sojin An, Javier Seravalli, Uhn-Soo Cho, Ruma V. Banerjee 2017 University of Michigan Medical School

Structural And Mechanistic Insights Into Hemoglobincatalyzed Hydrogen Sulfide Oxidation And The Fate Of Polysulfide Products, Victor Vitvitsky, Pramod K. Yadav, Sojin An, Javier Seravalli, Uhn-Soo Cho, Ruma V. Banerjee

Department of Biochemistry: Faculty Publications

Hydrogen sulfide is a cardioprotective signaling molecule but is toxic at elevated concentrations. Red blood cells can synthesize H2S but, lacking organelles, cannot dispose of H2S via the mitochondrial sulfide oxidation pathway. We have recently shown that at high sulfide concentrations, ferric hemoglobin oxidizes H2S to a mixture of thiosulfate and iron-bound polysulfides in which the latter species predominates. Here, we report the crystal structure of human hemoglobin containing low spin ferric sulfide, the first intermediate in heme-catalyzed sulfide oxidation. The structure provides molecular insights into why sulfide is susceptible to oxidation in human …


Short Carboxylic Acid-Carboxylate Hydrogen Bonds Can Have Fully Localized Protons, Jiusheng Lin, Edwin Pozharski, Mark A. Wilson 2017 University of Nebraska - Lincoln

Short Carboxylic Acid-Carboxylate Hydrogen Bonds Can Have Fully Localized Protons, Jiusheng Lin, Edwin Pozharski, Mark A. Wilson

Department of Biochemistry: Faculty Publications

Short hydrogen bonds (H-bonds) have been proposed to play key functional roles in several proteins. The location of the proton in short H-bonds is of central importance, as proton delocalization is a defining feature of low barrier hydrogen bonds (LBHBs). Experimentally determining proton location in H-bonds is challenging. Here, bond length analysis of atomic (1.15–0.98 Å) resolution X-ray crystal structures of the human protein DJ-1 and its bacterial homolog YajL was used to determine the protonation states of H-bonded carboxylic acids. DJ-1 contains a buried, dimer-spanning 2.49 Å H-bond between Glu15 and Asp23 that satisfies standard donor-acceptor distance criteria for …


Re-Framing Biotechnology Regulation, Alison Peck 2017 West Virginia University College of Law

Re-Framing Biotechnology Regulation, Alison Peck

Law Faculty Scholarship

Biotechnology is about to spill the banks of federal regulation. New genetic engineering techniques like CRISPR-Cas9 promise revolutionary breakthroughs in medicine, agriculture, and public health-but those techniques would not be regulated under the terms of the Coordinated Framework for Regulation of Biotechnology. This revolutionary moment in biotechnology offers an opportunity to correct the flaws in the framework, which was hastily patched together at the advent of the technology. The framework has never captured all relevant technologies, has never satisfied the public that risk is being effectively managed, and has never been accessible to small companies and publicly-funded labs that increasingly …


Bioactive Compounds From Ventilago Denticulata Willd., Wannapha Molee 2017 Faculty of Science

Bioactive Compounds From Ventilago Denticulata Willd., Wannapha Molee

Chulalongkorn University Theses and Dissertations (Chula ETD)

New naphthalene derivatives (1 and 2) and a new isomer of ventilagolin (3), together with three known anthraquinones, including chrysophanol (4), physcin or emodin 3-methyl ether (5), and emodin (6), were isolated from vines of Ventilago denticulata. The isolated compounds exhibited cytotoxic activity against cancer cell lines (HuCCA-1, A549, HepG-2, MOLT-3) with IC50 values of 1.15-40.54 µg/mL. Compounds 1-3 exhibited weak antibacterial activity (MIC 200.0-400.0 µg/mL), while emodin (6) displayed moderate antibacterial activity with MIC 25.0 µg/mL. Compounds 1-3, 5, and 6 exhibited nitric oxide radical scavenging activities with IC50 232.0-604.3 µM and compounds 1 and 2 exhibited DPPH radical …


Chaperonin Containing Tcp1 (Cct) As A Target For Cancer Therapy, Ana Carr 2017 University of Central Florida

Chaperonin Containing Tcp1 (Cct) As A Target For Cancer Therapy, Ana Carr

Electronic Theses and Dissertations

Treatments for aggressive cancers like triple negative breast cancer (TNBC) and small-cell lung cancer (SCLC) have not improved and remain associated with debilitating side effects. There is an unmet medical need for better, druggable targets and improved therapeutics. To this end, we investigated the role of Chaperonin-Containing TCP1 (CCT), an evolutionarily conserved protein-folding complex composed of eight subunits (CCT1-8), in oncogenesis. Our laboratory was the first to report that the CCT2 subunit is highly expressed in breast cancer and could be therapeutically targeted. To determine whether CCT is a marker of disease progression in other cancers, we analyzed CCT2 gene …


Fractionation And Characterization Of Lignin Streams From Genetically Engineered Switchgrass, Enshi Liu 2017 University of Kentucky

Fractionation And Characterization Of Lignin Streams From Genetically Engineered Switchgrass, Enshi Liu

Theses and Dissertations--Biosystems and Agricultural Engineering

Development of biomass feedstocks with desirable traits for cost-effective conversion is one of the main focus areas in biofuels research. As suggested by techno-economic analyses, the success of a lignocellulose-based biorefinery largely relies on the utilization of lignin to generate value-added products, i.e. fuels and chemicals. The fate of lignin and its structural/compositional changes during pretreatment have received increasing attention; however, the effect of genetic modification on the fractionation, depolymerization and catalytic upgrading of lignin from genetically engineered plants is not well understood. This study aims to fractionate and characterize the lignin streams from a wild-type and two genetically engineered …


Assessment Of Tattoo And Silicone Wounds In Terms Of Time Of Treatment And Perceived Treatment Quality, M Pettitt 2017 University of Central Florida

Assessment Of Tattoo And Silicone Wounds In Terms Of Time Of Treatment And Perceived Treatment Quality, M Pettitt

Electronic Theses and Dissertations

At the point of injury, critical medical tasks include locating and identifying an injury as well as applying the appropriate initial care. Over the past decade, to increase the fidelity of wound representation and ultimately the quality of medical care, a considerable amount of research and development has occurred to improve simulated wounds during training, primarily at the point of injury. As material and techniques mature and as more relevant data is collected on tissue properties, examining what fidelity is required for training at the point of injury is crucial. The main objective of this effort was to assess a …


Development Of Gene-Based Microsatellite Markers In Acer Saccharum Marsh., Monica Harmon 2017 Michigan Technological University

Development Of Gene-Based Microsatellite Markers In Acer Saccharum Marsh., Monica Harmon

Dissertations, Master's Theses and Master's Reports

Sugar maple (Acer saccharum Marsh) is an important ecological and economic resource. Despite its significance, there are few molecular genetic resources available to date. This study used previously developed EST libraries generated for sugar maple and other hardwood species to develop and characterize EST-SSRs in sugar maple and test their transferability to other Acer species. Out of 58 markers tested, 22 showed amplification in the expected size range. From these, 16 polymorphic markers were selected to test transferability and 13 of these showed amplification in at least one other Acer species.


Neutral And Adaptive Genetic Variation In North American Hardwood Tree Species, Sudhir Khodwekar 2017 Michigan Technological University

Neutral And Adaptive Genetic Variation In North American Hardwood Tree Species, Sudhir Khodwekar

Dissertations, Master's Theses and Master's Reports

'Northern Hardwoods,’ are a characteristic composition of multiple tree species and a part of the rich and diverse northeastern forests. Hardwoods e.g. oaks (Quercus: Fagaceae and Sugar maple), serve as foundation species and offer ecological and economical gains to wildlife and humans. Most of the forest trees go through many biotic and abiotic stresses, for example climate change in their long life span. To overcome these threats and to adapt to changing conditions, tree species need to maintain variation especially in adaptive genes. Therefore, it is necessary to analyze genetic diversity within forest tree populations and take appropriate …


The Role Of Alternative Polyadenylation Mediated By Cpsf30 In Arabidopsis Thaliana, Guijie Hao 2017 University of Kentucky

The Role Of Alternative Polyadenylation Mediated By Cpsf30 In Arabidopsis Thaliana, Guijie Hao

Theses and Dissertations--Plant and Soil Sciences

Drought stress is considered one of the most devastating abiotic stress factors that limit crop productivity for modern agriculture worldwide. There is a large range of physiological and biochemical responses induced by drought stress. The responses range from physiological and biochemical to regulation at transcription and posttranscriptional levels. Post-transcription, the products encoded by eukaryotic genes must undergo a series of modifications to become a mature mRNA. Polyadenylation is an important one in terms of regulation. Polyadenylation impacts gene expression through determining the coding and regulation potential of the mRNA, especially when different mRNAs from the same gene may be polyadenylated …


Transcriptional And Post-Translational Regulation Of Terpenoid Indole Alkaloid Biosynthesis In Catharanthus Roseus, Priyanka Paul 2017 University of Kentucky

Transcriptional And Post-Translational Regulation Of Terpenoid Indole Alkaloid Biosynthesis In Catharanthus Roseus, Priyanka Paul

Theses and Dissertations--Plant and Soil Sciences

Catharanthus roseus (Madagascar periwinkle) is the exclusive source of an array of terpenoid indole alkaloids (TIAs) that are used in the treatments of hypertension and certain types of cancer. TIA biosynthesis is under stringent spatiotemporal control and is induced by jasmonate (JA) and fungal elicitors. Tryptamine, derived from the indole branch, and secologanin from the iridoid branch are condensed to form the first TIA, strictosidine. Biosynthesis of TIA is regulated at the transcriptional level and several transcription factors (TFs) regulating the expression of genes encoding key enzymes in the pathway have been isolated and characterized. The JA-responsive APETALA2/ETHYLENE RESPONSE FACTOR …


An Undergraduate Cell Biology Lab: Western Blotting To Detect Proteins From Drosophila Eye, Neha Gogia, Ankita Sarkar, Amit Singh 2017 University of Dayton

An Undergraduate Cell Biology Lab: Western Blotting To Detect Proteins From Drosophila Eye, Neha Gogia, Ankita Sarkar, Amit Singh

Biology Faculty Publications

We have developed an undergraduate laboratory to allow detection and localization of proteins in the compound eye of Drosophila melanogaster, a.k.a fruit fly. This lab was a part of the undergraduate curriculum of the cell biology laboratory course aimed to demonstrate the use of Western Blotting technique to study protein localization in the adult eye of Drosophila. Western blotting, a two-day laboratory exercise, can be used to detect the presence of proteins of interests from total protein isolated from a tissue. The first day involves isolation of proteins from the tissue and SDS-PAGE (sodium dodecyl sulfate-polyacrylamide) gel …


Real-Time Quantitative Pcr To Demonstrate Gene Expression In An Undergraduate Lab, Abijeet Singh Mehta, Amit Singh 2017 University of Dayton

Real-Time Quantitative Pcr To Demonstrate Gene Expression In An Undergraduate Lab, Abijeet Singh Mehta, Amit Singh

Biology Faculty Publications

The objective of this teaching note is to develop a laboratory exercise, which allows students to get a hands-on experience of a molecular biology technique to analyze gene expression. The short duration of the biology laboratory for an undergraduate curriculum is the biggest challenge with the development of new labs. An important part of cell biology or molecular biology undergraduate curriculum is to study gene expression. There are many labs to study gene expression in qualitative manner. The commonly used reporter gene expression studies are primarily qualitative. However, there is no hands-on experience exercise to quantitatively determine gene expression. Therefore, …


Portulaca Oleracea Extract Can Inhibit Nodule Formation Of Colon Cancer Stem Cells By Regulating Gene Expression Of Notch Signal Transduction Pathway, Li Chen 2017 Michigan Technological University

Portulaca Oleracea Extract Can Inhibit Nodule Formation Of Colon Cancer Stem Cells By Regulating Gene Expression Of Notch Signal Transduction Pathway, Li Chen

Dissertations, Master's Theses and Master's Reports

The main aim of this study was to investigate the effects of Portulaca oleracea extract on tumor formation in colon cancer stem cells and chemotherapy sensitivity. In addition, this study analyzed the genetic changes within the Notch signal transduction pathway associated with the effects of the extracts. Serum-free cultures of colon cancer cells (HT-29) and HT-29 stem cells were treated with the chemotherapeutic drug 5-FU to assess sensitivity. Injections of the stem cells were also given to BALB/c mice to confirm tumor growth and characteristics. In addition, the effect of different concentrations of P. oleracea extract was tested on the …


Uv-A And -B Blocking Agents For Psoriasis Phototherapy, Sruangsuda Mawaha 2017 Faculty of Science

Uv-A And -B Blocking Agents For Psoriasis Phototherapy, Sruangsuda Mawaha

Chulalongkorn University Theses and Dissertations (Chula ETD)

UV-A and -B blocking agents are essential components for the formulation of topical cream for psoriasis phototherapy. In this research, many natural compounds were discovered as potent UV-A and UV-B blocking agents, but small number could absorb narrowband UV-B. Quercetin hydrate, oxyresveratrol and curcumin were good UV-A blocking agents, while catechin, hesperetin, lutein and naringenin were proper for UV-B blocking agents. Berberine chloride hydrate, demethoxycurcumin and mansonone G were UV-A and -B blocking agents. From random combination, the mixture of mansonone G 0.010 % w/v and quercetin hydrate 0.0025 % w/v was disclosed to be a right choice for this …


Bioactive Compounds With An Inhibitory Activity Against Osteoclast Differentiation And Mechanisms, Supatta Chawalitpong 2017 Faculty of Science

Bioactive Compounds With An Inhibitory Activity Against Osteoclast Differentiation And Mechanisms, Supatta Chawalitpong

Chulalongkorn University Theses and Dissertations (Chula ETD)

โรคกระดูกพรุนเป็นปัญหาสุขภาพที่สำคัญอย่างหนึ่งของสังคมผู้สูงอายุ สาเหตุหลักของภาวะดังกล่าวเกิดจากความไม่สมดุลของเซลล์ที่ทำหน้าทีสร้างกระดูก (osteoblast) และเซลล์ที่ทำหน้าที่สลายกระดูก (osteoclast) การลดลงของมวลกระดูกในผู้สูงอายุนำมาซึ่งความเสี่ยงในการแตกหักของกระดูก จุดประสงค์ของการศึกษาครั้งนี้ คือ การคัดกรองสารออกฤทธิ์ทางชีวภาพที่มีฤทธิ์ในการต้านการเปลี่ยนสภาพของเซลล์สลายกระดูกและกลไกการออกฤทธิ์ของสารนั้นๆ ในกลุ่มสารทดสอบอนุพันธ์สาร (3S)-1-(3,4-dihydroxyphenyl)-3-hydroxy-7-phenyl-(6E)-6-heptene (DHPH) และ กรดไซเปอเรโนอิก (Cyperenoic acid) ซึ่งสกัดได้จากส่วนเหง้า Curcuma comosa Rox. และราก Croton crassifolius Geiseler. ตามลำดับ มีฤทธิ์แรงในการต้านการเกิดเซลล์สลายกระดูก ในระดับ in vitro ใช้เซลล์จากไขกระดูกที่ถูกกระตุ้นด้วย receptor activator of nuclear factor-κB ligand (RANKL) DHPH มีฤทธิ์ในการยับยั้งการเปลี่ยนสภาพไปเป็นเซลล์สลายกระดูก โดยยับยั้งการเกิด TRAP-positive multinucleated cells (TRAP+ MNCs) ที่ระดับความเข้มข้นยับยั้งกึ่งหนึ่ง (IC50) 325±1.37 นาโนโมลาร์ การยับยั้งดังกล่าวสอดคล้องกับการลดต่ำลงของสัญญาณภายใต้การกระตุ้นของ RANKL โดยรวมถึงวิถี Mitogen-activated protein kinases (MAPKs) เช่น ERK (p44/42) และ p38 แต่ไม่มีผลต่อวิถี NF-κB โดยผลของสารดังกล่าวยังส่งผลกระทบต่อระดับของ transcription factor เช่น nuclear factor of activation T cells (NFATc1) และ c-Fos ซึ่งมีความสำคัญมากในการเปลี่ยนสภาพเป็นเซลล์สลายกระดูก DHPH ลดการกัดกร่อนกระดูก (bone resorption) บนแผ่นกระดูก (bone slices) นอกจากนั้นสาร DHPH ยังสามารถเพิ่มกิจกรรมของเอนไซม์ alkaline phosphatase (ALP) ที่เกี่ยวข้องกับการเปลี่ยนสภาพของเซลล์สร้างกระดูกในเซลล์ชนิด MC3T3-E1 ในกรณีของกรดไซเปอเรโนอิกนั้น พบว่าสามารถลดการเกิด TRAP+ MNCs ที่ระดับความเข้มข้น IC50 ที่ 36.69±1.02 …


Hyper-Activation Of Pp60(Src) Limits Nitric Oxide Signaling By Increasing Asymmetric Dimethylarginine Levels During Acute Lung Injury, Sanjiv Kumar, Xutong Sun, Satish Kumar Noonepalle, Qing Lu, Evgeny Zemskov, Ting Wang, Saurabh Aggarwal, Christine Gross, Shruti Sharma, Ankit A. Sesai, John D. Catravas 2017 Old Dominion University

Hyper-Activation Of Pp60(Src) Limits Nitric Oxide Signaling By Increasing Asymmetric Dimethylarginine Levels During Acute Lung Injury, Sanjiv Kumar, Xutong Sun, Satish Kumar Noonepalle, Qing Lu, Evgeny Zemskov, Ting Wang, Saurabh Aggarwal, Christine Gross, Shruti Sharma, Ankit A. Sesai, John D. Catravas

Bioelectrics Publications

The molecular mechanisms by which the endothelial barrier becomes compromised during lipopolysaccharide (LPS) mediated acute lung injury (ALI) are still unresolved. We have previously reported that the disruption of the endothelial barrier is due, at least in part, to the uncoupling of endothelial nitric oxide synthase (eNOS) and increased peroxynitrite-mediated nitration of RhoA. The purpose of this study was to elucidate the molecular mechanisms by which LPS induces eNOS uncoupling during ALI. Exposure of pulmonary endothelial cells (PAEC) to LPS increased pp60Src activity and this correlated with an increase in nitric oxide (NO) production, but also an increase in …


Analysis Of Microbial Diversity In Disturbed Soil, Tyler G. Sanda 2017 University of Akron

Analysis Of Microbial Diversity In Disturbed Soil, Tyler G. Sanda

Williams Honors College, Honors Research Projects

This paper uses the composition and abundance of microbial species to analyze soil recovery in disturbed land. Surface mining disturbs ecological communities throughout the world. As organizations seek to reclaim these disturbed lands, a proper analysis of recovery is needed. In previous studies, recovery of disturbed land was limited to surface examinations, which do not characterize the possible unseen devastating effects of the subsoil. Soil microorganisms are extremely sensitive to environmental changes such as strip mining. It is proposed that these microorganisms may serve as better indicators of recovery post disturbance. Our analysis indicates microbial recovery, however it may not …


Oxidative Stress, Metabolomics Profiling, And Mechanism Of Local Anesthetic Induced Cell Death In Yeast, Cory Honsinger Thomas Boone, Ryan A. Grove, Dana Adamcova, Javier Seravalli, Jiri Adamec 2017 University of Nebraska - Lincoln

Oxidative Stress, Metabolomics Profiling, And Mechanism Of Local Anesthetic Induced Cell Death In Yeast, Cory Honsinger Thomas Boone, Ryan A. Grove, Dana Adamcova, Javier Seravalli, Jiri Adamec

Department of Biochemistry: Faculty Publications

The World Health Organization designates lidocaine as an essential medicine in healthcare, greatly increasing the probability of human exposure. Its use has been associated with ROS generation and neurotoxicity. Physiological and metabolomic alterations, and genetics leading to the clinically observed adverse effects have not been temporally characterized. To study alterations that may lead to these undesirable effects, Saccharomyces cerevisiae grown on aerobic carbon sources to stationary phase was assessed over 6 h. Exposure of an LC50 dose of lidocaine, increased mitochondrial depolarization and ROS/RNS generation assessed using JC-1, ROS/RNS specific probes, and FACS. Intracellular calcium also increased, assessed by …


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