Biotechnology Commons^™

Cell Fragmentation And Permeabilization By A 1 Ns Pulse Driven Triple-Point Electrode, Enbo Yang, Joy Li, Michael Cho, Shu Xiao

Bioelectrics Publications

Ultrashort electric pulses (ns-ps) are useful in gaining understanding as to how pulsed electric fields act upon biological cells, but the electric field intensity to induce biological responses is typically higher than longer pulses and therefore a high voltage ultrashort pulse generator is required. To deliver 1 ns pulses with sufficient electric field but at a relatively low voltage, we used a glass-encapsulated tungsten wire triple-point electrode (TPE) at the interface among glass, tungsten wire, and water when it is immersed in water. A high electric field (2MV/cm) can be created when pulses are applied. However, such a high electric …

Upregulation Of Dna Sensors In B16.F10 Melanoma Spheroid Cells After Electrotransfer Of Pdna, Katarina Znidar, Masa Bosnjak, Tanja Jesenko, Loree C. Heller, Maja Cemazar

Bioelectrics Publications

Increased expression of cytosolic DNA sensors, a category of pattern recognition receptor, after control plasmid DNA electrotransfer was observed in our previous studies on B16.F10 murine melanoma cells. This expression was correlated with the upregulation of proinflammatory cytokines and chemokines and was associated with cell death. Here, we expanded our research to include the influence of features of cells in a 3-dimensional environment, which better represents the tumors’ organization in vivo. Our results show that lower number of cells were transfected in spheroids compared to 2-dimensional cultures, that growth was delayed after electroporation alone or after electrotransfer of plasmid …

Excitation And Injury Of Adult Ventricular Cardiomyocytes By Nano- To Millisecond Electric Shocks, Iurii Semenov, Sergey Grigoryev, Johanna U. Neuber, Christian W. Zemlin, Olga N. Pakhomova, Maura Casciola, Andrei G. Pakhomov

Bioelectrics Publications

Intense electric shocks of nanosecond (ns) duration can become a new modality for more efficient but safer defibrillation. We extended strength-duration curves for excitation of cardiomyocytes down to 200 ns, and compared electroporative damage by proportionally more intense shocks of different duration. Enzymatically isolated murine, rabbit, and swine adult ventricular cardiomyocytes (VCM) were loaded with a Ca²⁺ indicator Fluo-4 or Fluo-5N and subjected to shocks of increasing amplitude until a Ca²⁺ transient was optically detected. Then, the voltage was increased 5-fold, and the electric cell injury was quantified by the uptake of a membrane permeability marker dye, propidium …

Nano-Pulse Stimulation For The Treatment Of Pancreatic Cancer And The Changes In Immune Profile, Sigi Guo, Niculina I. Burcus, James Hornef, Yu Jing, Chunqi Jiang, Richard Heller, Stephen J. Beebe

Bioelectrics Publications

A Pancreatic cancer is a notorious malignant neoplasm with an extremely poor prognosis. Current standard of care is rarely effective against late-stage pancreatic cancer. In this study, we assessed nanopulse stimulation (NPS) as a local treatment for pancreatic cancer in a syngeneic mouse Pan02 pancreatic cancer model and characterized corresponding changes in the immune profile. A single NPS treatment either achieved complete tumor regression or prolonged overall survival in animals with partial tumor regression. While this is very encouraging, we also explored if this local ablation effect could also result in immune stimulation, as was observed when NPS led to …

Dielectric Properties Of Isolated Adrenal Chromaffin Cells Determined By Microfluidic Impedance Spectroscopy, A. C. Sabuncu, M. Stacey, G. L. Graviso, N. Semenova, P. T. Vernier, N. Leblanc, I. Chatterjee, J. Zaklit

Bioelectrics Publications

Knowledge of the dielectric properties of biological cells plays an important role in numerical models aimed at understanding how high intensity ultrashort nanosecond electric pulses affect the plasma membrane and the membranes of intracellular organelles. To this end, using electrical impedance spectroscopy, the dielectric properties of isolated, neuroendocrine adrenal chromaffin cells were obtained. Measured impedance data of the cell suspension, acquired between 1 kHz and 20 MHz, were fit into a combination of constant phase element and Cole-Cole models from which the effect of electrode polarization was extracted. The dielectric spectrum of each cell suspension was fit into a Maxwell-Wagner …

Photosystem Ii Subunit S Overexpression Increases The Efficiency Of Water Use In A Field-Grown Crop, Katarzyna Glowacka, Johannes Kromdijk, Katherine Kucera, Jiayang Xie, Amanda P. Cavanagh, Lauriebeth Leonelli, Andrew D. B. Leakey, Donald R. Ort, Krishna K. Niyogi, Stephen P. Long

Department of Biochemistry: Faculty Publications

Insufficient water availability for crop production is a mounting barrier to achieving the 70% increase in food production that will be needed by 2050. One solution is to develop crops that require less water per unit mass of production. Water vapor transpires from leaves through stomata, which also facilitate the influx of CO2 during photosynthetic assimilation. Here, we hypothesize that Photosystem II Subunit S (PsbS) expression affects a chloroplastderived signal for stomatal opening in response to light, which can be used to improve wateruse efficiency. Transgenic tobacco plants with a range of PsbS expression, from undetectable to 3.7 …

Simulation Of Stimulation: Cytokine Dosage And Cell Cycle Crosstalk Driving Timing-Dependent T Cell Differentiation, Matteo Barberis, Tomáš Helikar, Paul Verbruggen

Department of Biochemistry: Faculty Publications

Triggering an appropriate protective response against invading agents is crucial to the effectiveness of human innate and adaptive immunity. Pathogen recognition and elimination requires integration of a myriad of signals from many different immune cells. For example, T cell functioning is not qualitatively, but quantitatively determined by cellular and humoral signals. Tipping the balance of signals, such that one of these is favored or gains advantage on another one, may impact the plasticity of T cells. This may lead to switching their phenotypes and, ultimately, modulating the balance between proliferating and memory T cells to sustain an appropriate immune response. …

Computational Systems Biology Approach For The Study Of Rheumatoid Arthritis: From A Molecular Map To A Dynamical Model, Vidisha Singh, Marek Ostaszewski, George D. Kalliolias, Gilles Chiocchia, Robert Olaso, Elisabeth Petit-Teixeira, Tomáš Helikar, Anna Niarakis

Department of Biochemistry: Faculty Publications

In this work we present a systematic effort to summarize current biological pathway knowledge concerning Rheumatoid Arthritis (RA). We are constructing a detailed molecular map based on exhaustive literature scanning, strict curation criteria, re-evaluation of previously published attempts and most importantly experts’ advice. The RA map will be web-published in the coming months in the form of an interactive map, using the MINERVA platform, allowing for easy access, navigation and search of all molecular pathways implicated in RA, serving thus, as an on line knowledgebase for the disease. Moreover the map could be used as a template for Omics data …

The Colomoto Interactive Notebook: Accessible And Reproducible Computational Analyses For Qualitative Biological Networks, Aurelien Naldi, Celine Hernandez, Nicolas Levy, Gautier Stoll, Pedro T. Monteiro, Claudine Chaouiya, Tomáš Helikar, Andrei Zinovyev, Laurence Calzone, Sarah Cohen-Boulakia, Denis Thieffry, Loic Pauleve

Department of Biochemistry: Faculty Publications

Analysing models of biological networks typically relies on workflows in which different software tools with sensitive parameters are chained together,many times with additional manual steps. The accessibility and reproducibility of such workflows is challenging, as publications often overlook analysis details, and because some of these tools may be difficult to install, and/or have a steep learning curve. The CoLoMoTo Interactive Notebook provides a unified environment to edit, execute, share, and reproduce analyses of qualitative models of biological networks. This framework combines the power of different technologies to ensure repeatability and to reduce users’ learning curve of these technologies. The framework …

A Mechanistic Computational Model Reveals That Plasticity Of Cd4+ T Cell Differentiation Is A Function Of Cytokine Composition And Dosage, Bhanwar Lal Puniya, Robert G. Todd, Akram Mohammed, Deborah M. Brown, Matteo Barberis, Tomáš Helikar

Department of Biochemistry: Faculty Publications

CD4+ T cells provide cell-mediated immunity in response to various antigens. During an immune response, naïve CD4+ T cells differentiate into specialized effector T helper (Th1, Th2, and Th17) cells and induced regulatory (iTreg) cells based on a cytokine milieu. In recent studies, complex phenotypes resembling more than one classical T cell lineage have been experimentally observed. Herein, we sought to characterize the capacity of T cell differentiation in response to the complex extracellular environment. We constructed a comprehensive mechanistic (logical) computational model of the signal transduction that regulates T cell differentiation. The model’s dynamics were characterized and analyzed under …

Reactive Oxygen Species In Chronic Obstructive Pulmonary Disease, Samia Boukhenouna, Mark A. Wilson, Karim Bahmed, Beata Kosmider

Department of Biochemistry: Faculty Publications

Chronic obstructive pulmonary disease (COPD) includes chronic bronchitis and emphysema. Environmental exposure, primarily cigarette smoking, can cause high oxidative stress and is the main factor of COPD development. Cigarette smoke also contributes to the imbalance of oxidant/antioxidant due to exogenous reactive oxygen species (ROS). Moreover, endogenously released ROS during the inflammatory process and mitochondrial dysfunction may contribute to this disease progression. ROS and reactive nitrogen species (RNS) can oxidize different biomolecules such as DNA, proteins, and lipids leading to epithelial cell injury and death. Various detoxifying enzymes and antioxidant defense systems can be involved in ROS removal. In this review, …

Electrotransfer Of Different Control Plasmids Elicits Different Antitumor Effectiveness In B16.F10 Melanoma, Masa Bosnjak, Tanjo Jesenko, Urska Kamensek, Gregor Sersa, Jaka Lavrencak, Loree Heller, Maja Cemazar

Bioelectrics Publications

Several studies have shown that different control plasmids may cause antitumor action in different murine tumor models after gene electrotransfer (GET). Due to the differences in GET protocols, plasmid vectors, and experimental models, the observed antitumor effects were incomparable. Therefore, the current study was conducted comparing antitumor effectiveness of three different control plasmids using the same GET parameters. We followed cytotoxicity in vitro and the antitumor effect in vivo after GET of control plasmids pControl, pENTR/U6 scr and pVAX1 in B16.F10 murine melanoma cells and tumors. Types of cell death and upregulation of selected cytosolic DNA sensors and cytokines were …

การปรับปรุงกระบวนการสกัดและการทำให้บริสุทธิ์สำหรับพอลิ 3-ไฮดรอกซีบิวทิเรตจาก Cupriavidus Necator สายพันธุ์ A-04 โดยใช้ 1,3-ไดออกโซเลน, ชนากานต์ วงศ์มูล

Chulalongkorn University Theses and Dissertations (Chula ETD)

พอลิไฮดรอกซีบิวทิเรต (polyhydroxybutyrate, PHB) เป็นพอลิเอสเทอร์ชีวภาพที่ถูกสร้างและสะสมอยู่ภายในเซลล์แบคทีเรียในรูปแบบแกรนูลภายในเซลล์ เทคนิคหลายประการได้รับการพัฒนาสำหรับขั้นตอนการสกัดและการทำบริสุทธิ์ให้มีประสิทธิภาพ ราคาไม่แพง และเป็นมิตรต่อสิ่งแวดล้อม ในการศึกษานี้ใช้ 1,3-ไดออกโซเลน ซึ่งตัวทำละลายที่เป็นมิตรต่อสิ่งแวดล้อม ในการสกัด PHB ร่วมกับวิธีต่างๆ ซึ่งรวมถึงการใช้ 1,3-ไดออกโซเลน เจือจาง การสกัดด้วยเครื่องซอกห์เลต การสกัดด้วยวิธีเชิงกลร่วมกับตัวทำละลาย และการเขย่าต่อเนื่องระหว่างสกัด ผลการทดลองพบว่าการเพิ่มอัตราการเขย่าในระหว่างกระบวนการสกัดอาจเพิ่มเปอร์เซ็นต์การนำกลับและความบริสุทธิ์ การสกัด PHB จากเซลล์แห้งของ Cupriavidus necator สายพันธุ์ A-04 ซึ่งมีการสะสม PHB เท่ากับ 70.23±1.08 เปอร์เซ็นต์ต่อน้ำหนักเซลล์แห้ง ที่อุณหภูมิ 80 องศาเซลเซียส อัตราการเขย่า 150 รอบต่อนาที เป็นเวลา 5 ชั่วโมง เป็นผลให้การนำกลับ PHB เท่ากับ 86.59±2.44 เปอร์เซ็นต์ และความบริสุทธิ์ 99.03±0.22 เปอร์เซ็นต์ การสกัด PHB จากเซลล์เปียกภายใต้ภาวะเดียวกันเป็นผลให้การนำกลับและความบริสุทธิ์เท่ากับ 84.63±2.17 และ 97.92±0.26 เปอร์เซ็นต์ ตามลำดับ เมื่อขยายระดับการสกัด PHB จากเซลล์แห้งที่ระดับการสกัด 1 ลิตร ให้ความบริสุทธิ์สูงถึง 99.33±0.54 เปอร์เซ็นต์ และการนำกลับเท่ากับ 64.05±0.35 เปอร์เซ็นต์ น้ำหนักโมเลกุล สมบัติเชิงความร้อน และสมบัติเชิงกลของ PHB ที่ได้จากการสกัดด้วย 1,3-ไดออกโซเลน ใกล้เคียงกับ PHB ที่ได้จากการสกัดด้วยคลอโรฟอร์ม ดังนั้น การเขย่า อุณหภูมิ และภาวะของเซลล์ที่ใช้ในการสกัด (เซลล์แห้งหรือเซลล์เปียก) เป็นปัจจัยสำคัญในการพัฒนาวิธีให้มีประสิทธิภาพ อย่างไรก็ตามการนำ 1,3-ไดออกโซเลน ที่ใช้แล้วกลับมาใช้ใหม่นั้นให้เปอร์เซ็นต์การนำกลับและความบริสุทธิ์ต่ำไม่คุ้มค่ากับพลังงานที่เสียไปในกระบวนการแยกสารเพื่อนำกลับมาใช้ใหม่

Plant Probiotic Bacteria Bacillus And Paraburkholderia Improve Growth, Yield And Content Of Antioxidants In Strawberry Fruit, Mosaddiqur Rahman, Abdullah As Sabir, Julakha Akter Mukrta, Mohibul Alam Khan, Mohammed Mohi-Ud-Din, Giashuddin Miah, Mahfuzur Rahman, M. Tofazzal Islam

Faculty & Staff Scholarship

Strawberry is an excellent source of natural antioxidants with high capacity of scavenging free radicals.

This study evaluated the effects of two plant probiotic bacteria, Bacillus amylolequefaciens BChi1 and Paraburkholderia fungorum BRRh-4 on growth, fruit yield and antioxidant contents in strawberry fruits. Root dipping of seedlings (plug plants) followed by spray applications of both probiotic bacteria in the field on foliage significantly increased fruit yield (up to 48%) over non-treated control. Enhanced fruit yield likely to be linked with higher root and shoot growth, individual and total fruit weight/plant and production of phytohormone by the probiotic bacteria applied on plants. …

Research Of Sustainable Jet Fuel Production Using Microbes, Rajee Olaganathan

Publications

Global climate change, coupled with rapidly increasing oil prices and energy demand around the world, has paved a way for intense research in the biofuel sector. Stakeholders in the aviation industry have started to focus on bio-jet fuel. Bio-jet fuel is regarded as a sustainable solution to greenhouse gas emissions and energy demand. This paper provides a brief review of the biofuel production technologies, the role of bacteria in producing hydrocarbons and the recent advancements in microbial engineering to enhance the biofuel production. Finally, this paper concludes by highlighting the challenges and future research implications in bio-jet fuel production.

Prospective Study Of The Feasibility Of Point-Of-Care Testing Strategy For Carbapenem-Resistant Organism Detection, Rahul Pannala, Bruce Baldwin, Vijay Aluru, Thomas E. Grys, Jordan Holmes, Laurence J. Miller, M. Edwyn Harrison, Cuong C. Nguyen, Fred C. Tenover, David Persing, Douglas O. Faigel

Biology Faculty Publications

Background/aims: In an investigator-initiated, prospective study, we evaluated the feasibility of a five-gene sequence point-of-care (POC) testing strategy (Xpert CARBA-R Assay, Cepheid Inc., Sunnyvale, CA, USA), compared to reference laboratory PCR (48 – 72 hours turnaround time, two gene sequences), in patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) and in a hospital outbreak investigation.

Methods: After informed consent, patients undergoing ERCP (September 2015 – April 2016, n = 191) at Mayo Clinic and potential hospital contacts (n = 9) of an index carbapenem-resistant organism (CRO)-positive inpatient were included. Two rectal swabs, one each for reference and POC assays were obtained. The …

Range Expansion And The Origin Of Usa300 North American Epidemic Methicillin-Resistant Staphylococcus Aureus, Lavanya Challagundla, Isabella A. Tickler, Xavier Didelot, David C. Coleman, Anna C. Shore, Geoffrey W. Coombs, Daniel O. Sordelli, Eric L. Brown, Robert Skov, Anders Rhod Larsen, Jinnethe Reyes, Iraida E. Robledo, Guillermo J. Vazquez, Paul Rivera, Paul D. Fey, Kurt Stevenson, Shu-Hua Wang, Barry N. Kreiswirth, Jose R. Mediavilla, Cesar A. Arias, Paul J. Planet, Rathel L. Nolan, Fred C. Tenover, Richard V. Goering, D. Ashley Robinson

Biology Faculty Publications

The USA300 North American epidemic (USA300-NAE) clone of methicillin-resistant Staphylococcus aureus has caused a wave of severe skin and soft tissue infections in the United States since it emerged in the early 2000s, but its geographic origin is obscure. Here we use the population genomic signatures expected from the serial founder effects of a geographic range expansion to infer the origin of USA300-NAE and identify polymorphisms associated with its spread. Genome sequences from 357 isolates from 22 U.S. states and territories and seven other countries are compared. We observe two significant signatures of range expansion, including decreases in genetic diversity …

Building Capacity For A Global Genome Editing Observatory: Institutional Design, Krishanu Saha, J. Benjamin Hurlbut, Sheila Jasanoff, Aziza Ahmed, Anthony Appiah, Elizabeth Bartholet, Francoise Baylis, Gaymon Bennett, George Church, I. Glenn Cohen, George Daley, Kevin Finneran, William Hurlbut, Rudolf Jaenisch, Laurence Lwoff, John Paul Kimes, Peter Mills, Jacob Moses, Buhm Soon Park, Erik Parens, Rachel Salzman, Abha Saxena, Hilton Simmet, Tania Simoncelli, O. Carter Snead, Kaushik Sunder Rajan, Robert D. Truog, Patricia Williams, Christiane Woopen

Journal Articles

A new infrastructure is urgently needed at the global level to facilitate exchange on key issues concerning genome editing. We advocate the establishment of a global observatory to serve as a center for international, interdisciplinary, and cosmopolitan reflection.

Optimisation Of Ion Exchange Chromatography Purification Protocols For A Staphylococcal Peptidoglycan Degrading Hydrolase Enzyme, Fiona Maher

Theses

Bacteriophage (phage) are the most abundant biological entities on earth and were first discovered by d’Herelle in 1917. They are found wherever their hosts live and, like all viruses they do not have the ability to make their own protein. Therefore, in order to reproduce, phage must invade and infect bacterial cells. This project focused on the optimisation of Ion Exchange Chromatography purification protocols for a staphylococcal peptidoglycan degrading hydrolase enzyme (CHAPk). The project objective was to obtain the greatest yield of enzyme from the growth of the E.coU XL 1-Blue expression system into which the vector pQE60 was previously …

Role Of Single Nucleotide Polymorphisms (Snps) In Ptpn2/22 And Mycobacterium Avium Subspecies Paratuberculosis (Map) In Rheumatoid Arthritis And Crohn's Disease, Robert Sharp

Electronic Theses and Dissertations

Both genetic pre-disposition and potential environmental triggers are shared between Rheumatoid arthritis (RA) and Crohn's disease (CD). We hypothesized that single nucleotide polymorphisms (SNPs) in the negative T-cell regulators Protein Tyrosine Phosphatase Nonreceptor type 2 and 22 (PTPN2/22) lead to a dysregulated immune response as seen in RA and CD. To test the hypothesis, peripheral leukocytes samples from 204 consented subjects were TaqMan genotyped for 9 SNPs in PTPN2/22. The SNPs effect on PTPN2/22 and IFN-y expression was determined using RT-PCR. Blood samples were analyzed for the Mycobacterium avium subspecies paratuberculosis (MAP) IS900 gene by nPCR. T-cell proliferation and response …