Biotechnology Commons^™

Optimizing Gdna Purification And Qpcr Techniques For Specific Detection And Enumeration Of Probiotic Bacteria, Katelynn C. Lee, Jennifer Vanderkelen, Christopher Kitts

STAR Program Research Presentations

Probiotics are live microorganisms, like bacteria and yeast, which are thought to provide health benefits when ingested. Large manufacturers produce blends of probiotics for supplementing the diets of agriculturally important animals. Commercial probiotic products are labeled with the number of cells present at the time of manufacture. Our assignment was to develop an assay which can be used to quantify the number of specific lactic acid bacteria present in certain probiotic products, thus verifying the number reported on the product labels. This quantification process involves isolating genomic DNA (gDNA) from the given samples and then running the DNA through a …

Pupuk Anorganik Sebagai Alternatif Media Untuk Pertumbuhan Eksplan Nilam (Pogostemon Cablin Benth) Kultivar Sidikalang Dan Tapaktuan Secara In-Vitro, Suseno Amien, M. F. Wiguna

Jurnal Kultivasi

Kultur jaringan selama ini dipahami oleh sebagian masyarakat merupakan suatu teknologi yang mahal, terutama dalam menyediakan bahan kimia untuk media. Hasil percobaan sebelumnya menunjukkan bahwa percobaan dengan media pupuk anorganik dapat digunakan sebagi media tanam in-vitro. Namun media tersebut belum diteliti secara rinci manfaatnya untuk tanaman nilam. Penelitian ini dilakukan untuk memperoleh media pupuk anorganik yang sesuai untuk pertumbuhan kultivar nilam (Pogostemon cablin Benth) Sidikalang dan Tapak Tuan. Penelitian dilakukan di laboratorium teknologi kultur jaringan Pemuliaan Tanaman Fakultas Pertanian Universitas Padjadjaran dari bulan Mei sampai Agustus 2010. Rancangan yang digunakan adalah rancangan acak lengkap pola faktorial dengan dua faktor perlakuan …

Pengujian Efektivitas Berbagai Jenis Dan Konsentrasi Sitokinin Terhadap Multiplikasi Tunas Mikro Pisang (Musa Paradisiaca L.) Secara In Vitro, Bella Dian Ratnasari., Erni Suminar, Anne Nuraini, Ade Ismail

Jurnal Kultivasi

Perbanyakan tanaman pisang secara konvensional dengan menggunakan bonggol atau anakan (sucker) secara terus menerus dapat menurunkan kualitas dan memerlukan waktu lama serta meningkatkan terjadinya serangan penyakit. Oleh karena itu, suatu upaya untuk menghasilkan bibit yang memiliki kualitas baik dalam waktu yang cepat dapat menggunakan kultur in vitro. Percobaan ini bertujuan untuk memperoleh laju multiplikasi yang tinggi dengan penggunaan salah satu jenis dan konsentrasi sitokinin pada kultur in vitro tanaman pisang kepok kuning. Percobaan dilakukan pada bulan Oktober hingga Desember 2015 di Laboratorium Kultur Jaringan Teknologi Benih Fakultas Pertanian Universitas Padjadjaran. Rancangan percobaan menggunakan Rancangan Acak Lengkap (RAL) yang terdiri dari …

Contribution Of A Putative Up Element Dna Sequence To The Activity Of A Newly Identified Phage Promoter, Courtney Hamilton

Mahurin Honors College Capstone Experience/Thesis Projects

In transcription, a universal step in gene expression, information from a DNA sequence is copied into RNA. A key component in gene expression is the promoter sequence, a region of DNA to which RNA polymerase binds during the initiation of transcription of downstream genes. Most bacterial promoters contain a -10 and a -35 sequence that are bound by the RNA polymerase. Some promoters also contain an Upstream Promoter (UP) element. UP elements have been shown to boost promoter activity. We recently identified a new promoter in a mutant bacteriophage that grows on a bacterial host that prevents antitermination of phage …

Design And Development Of A Plasmid Vector For Protein Expression And Purification, Mahima Grover, Craig Sweet, David H. Thompson

The Summer Undergraduate Research Fellowship (SURF) Symposium

Production and isolation of proteins are difficult, costly and time-consuming processes. The aim of this project is for the development of plasmids, which allow for streamlined production and isolation of proteins. To allow for modular insertion of varying segments of DNA we are using ‘recursive directional ligation by plasmid reconstruction’. This technique uses type II restriction endonucleases, which cut downstream from their recognition site allowing multiple insertions without losing a restriction site. Using this process, we can ligate multiple DNA sequences together and express them to be able to construct a scar less fusion protein. In order to accomplish this, …

The Effect Of An Enhanced Isopentenyl Monophosphate Pool On Terpenoid Biosynthesis In Vivo, Evan T. Adams, Laura K. Henry, Dr. Natalia Dudareva

The Summer Undergraduate Research Fellowship (SURF) Symposium

Found in all living organisms, terpenoids make up the largest group of natural products and are essential compounds for many major processes, including photosynthesis, respiration, hormone production, and electron transport. Additionally, they have commercial and medical value in products including fragrances, cosmetics, and medicines. Terpenoids originate from the five-carbon building blocks isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are synthesized by the mevalonic acid (MVA) and methylerithritol phosphate (MEP) pathways. An alternative MVA pathway was discovered in Archaea with the final two enzymes being phosphomevalonate decarboxylase (MPD) and isopentenyl phosphate kinase (IPK). Even though this alternative pathway is not …

Optimization Of A Genomic Editing System Using Crispr/Cas9-Induced Site-Specific Gene Integration, Jillian L. Mccool Ms., Nick Hum, Gabriela G. Loots

STAR Program Research Presentations

The CRISPR-Cas system is an adaptive immune system found in bacteria which helps protect against the invasion of other microorganisms. This system induces double stranded breaks at precise genomic loci (1) in which repairs are initiated and insertions of a target are completed in the process. This mechanism can be used in eukaryotic cells in combination with sgRNAs (1) as a tool for genome editing. By using this CRISPR-Cas system, in addition to the “safe harbor locus,” ROSAβ26, the incorporation of a target gene into a site that is not susceptible to gene silencing effects can be achieved through few …

Vascular Endothelial Growth Factor-A Gene Electrotransfer Promotes Angiogenesis In A Porcine Model Of Cardiac Ischemia, Anna A. Bulysheva, Barbara Hargrave, Nina Burcus, Cathryn G. Lundberg, Len Murray, Richard Heller

Bioelectrics Publications

This study aimed to assess safety and therapeutic potential of gene electrotransfer (GET) as a method for delivery of plasmid encoding vascular endothelial growth factor A (VEGF-A) to ischemic myocardium in a porcine model. Myocardial ischemia was induced by surgically occluding the left anterior descending coronary artery in swine. GET following plasmid encoding VEGF-A injection was performed at four sites in the ischemic region. Control groups either received injections of the plasmid without electrotransfer or injections of the saline vehicle. Animals were monitored for 7 weeks and the hearts were evaluated for angiogenesis, myocardial infarct size and left ventricular contractility. …

Loss Of Exogenous Androgen Dependence By Prostate Tumor Cells Is Associated With Elevated Glucuronidation Potential, Brenna M. Zimmer, Michelle E. Howell, Qin Wei, Linlin Ma, Trevor Romsdahl, Eileen G. Loughman, Jennifer E. Markham, Javier Seravalli, Joseph J. Barycki, Melanie A. Simpson

Department of Biochemistry: Faculty Publications

Prostate epithelial cells control the potency and availability of androgen hormones in part by inactivation and elimination. UDP-glucose dehydrogenase (UGDH) catalyzes the NAD⁺-dependent oxidation of UDP-glucose to UDP-glucuronate, an essential precursor for androgen inactivation by the prostate glucuronidation enzymes UGT2B15 and UGT2B17. UGDH expression is androgen stimulated, which increases the production of UDP-glucuronate, and fuels UGT-catalyzed glucuronidation. In this study, we compared the glucuronidation potential and its impact on androgen-mediated gene expression in an isogenic LNCaP model for androgen dependent versus castration resistant prostate cancer. Despite significantly lower androgen-glucuronide output, LNCaP 81 castration resistant tumor cells expressed higher …

Novel Mechanisms Of Β-Adrenergic Signaling In Prostate Cancer Progression, Mohit Hulsurkar

Dissertations and Theses (Open Access)

Prostate cancer is the second leading cause of cancer death among American men. The American Cancer Society estimates that 180,890 men will be will be diagnosed with prostate cancer in 2016 in the USA. (http://www.cancer.org/cancer/prostatecancer/detailedguide/prostate-cancer-key-statistics). Androgen deprivation therapy (ADT) is the standard treatment for early stage prostate cancer. But most patients relapse with aggressive variants of prostate cancer, with survival time between 1-3 years. In order to develop cure for such aggressive variants of prostate cancer, our present understanding of the mechanisms underlying its progression needs to be advanced.

Recently, it has been found that activation of β-adrenergic signaling pathway …

Assessing Stress Responses To Atmospheric Cold Plasma Exposure Using Escherichia Coli Knock Out Mutants, Lu Han, Daniela Boehm, Sonal Patil, Patrick J. Cullen, Paula Bourke

Articles

Aim: This study investigated the effect of Atmospheric cold plasma (ACP) exposure induced stress on microbial inactivation patterns and the regulation of genes involved in the microbial stress response in conjunction with key processing parameters of exposure time and post treatment storage time.

Methods and Results: Cell suspensions of Escherichia coli BW 25113 and its isogenic knock-out mutants in rpoS, soxR, soxS, oxyR and dnaK genes were treated with high voltage ACP in a sealed package for 1, 3 and 5 min followed by 0, 1 and 24 h post-treatment storage. ROS densities and colony formation were …

Microbubble Generation By Piezoelectric Transducers For Biomedical Studies, Mohammed Alkhazal

Electrical & Computer Engineering Theses & Dissertations

Bubbles induced by blast waves or shocks are speculated as the major cause of damage in biological cells in mild traumatic brain injuries (TBI). Microbubble collapse was found to induce noticeable cell detachment from the cell substrate, changes in focal adhesion, and biomechanics. To better understand the bubble mechanism, a system needs to be constructed which allows clear differentiation on the impact of bubbles from that of shocks. Such a generator needs to be low profile in order to place under a microscope. A piezoelectric transducer system was designed to meet the need. The system uses either a flat or …

Small-Angle X-Ray Scattering Studies Of The Oligomeric State And Quaternary Structure Of The Trifunctional Proline Utilization A (Puta) Flavoprotein From Escherichia Coli, Ranjan K. Singh, John D. Larson, Weidong Zhu, Robert P. Rambo, Greg L. Hura, Donald F. Becker, John J. Tanner

Department of Biochemistry: Faculty Publications

Background: Trifunctional proline utilization A (PutA) proteins are multifunctional flavoproteins that catalyze two reactions and repress transcription of the put regulon.

Results: PutA from Escherichia coli is a V-shaped dimer, with the DNA-binding domain mediating dimerization.

Conclusion: Oligomeric state and quaternary structures are not conserved by PutAs.

Significance: The first three-dimensional structural information for any trifunctional PutA is reported.

Effect Of Rapamycin As An Inhibitor Of The Mtor Cell Cycle Entry Complex On The Selective Lysis Of Human Leukemia Cells Lines In Vitro Using 20 Khz Pulsed Low-Frequency Ultrasound, Yasmine Hemida

Renée Crown University Honors Thesis Projects - All

It has been shown that the mammalian target of rapamycin (mTOR) pathway, which regulates cell growth and proliferation, is aberrant in many hematological malignancies. Rapamycin inhibits mTOR signaling which regulates cell growth and cell cycle progression. This study sought to determine the effects of mTOR inhibitors on the cell sizes of normal and neoplastic cells and to determine ultrasonic sensitivity of normal and neoplastic cells treated with mTOR inhibitors. The effects of rapamycin (rapa), an extensively studied natural product that affects cell cycle entry by inhibiting mTORC1, and its analogs temsirolimus (tems) and everolimus (eve), were examined on leukemic cell …

Loss Of Cell Adhesion Increases Tumorigenic Potential Of Polarity Deficient Scribble Mutant Cells, Indrayani Waghmare, Madhuri Kango-Singh

Biology Faculty Publications

Epithelial polarity genes are important for maintaining tissue architecture, and regulating growth. The Drosophila neoplastic tumor suppressor gene scribble (scrib) belongs to the basolateral polarity complex. Loss of scrib results in disruption of its growth regulatory functions, and downregulation or mislocalization of Scrib is correlated to tumor growth. Somatic scribble mutant cells (scrib-) surrounded by wild-type cells undergo apoptosis, which can be prevented by introduction of secondary mutations that provide a growth advantage. Using genetic tools in Drosophila, we analyzed the phenotypic effects of loss of scrib in different growth promoting backgrounds. We investigated if a central …

Decision-Making Processes For A Patient Admitted To The Accident And Emergency Department Requiring Emergency Abdominal X-Ray ― A Case Study, Yuliawati Y.

ASEAN Journal on Science and Technology for Development

This study investigated the application of the various processes involved in decision-making for acase requiring radiologic emergency abdominal X-ray examination to the Accident and Emergency Department in a government-owned hospital in West Java, Indonesia. It is also aimed to evaluate the patient who was experiencing abdominal pain accompanied by bloating, vomiting, severe constipation and abdominal cramps.The decision-making models used for this case were the normative, prescriptive and descriptive judgments. A decision tree in the form of decision options that was employed in this case addressed the normative model of decision-making, while the algorithm and information processing theory focused on the …

Secretome Data From Trichoderma Reesei And Aspergillus Niger Cultivated In Submerged And Sequential Fermentation Methods, Camila Florencio, Fernanda M. Cunha, Alberto C. Badino, Cristiane S. Farinas, Eduardo Ximenes, Michael R. Ladisch

School of Agricultural & Biological Engineering Faculty Publications

The cultivation procedure and the fungal strain applied for enzyme production may influence levels and profile of the proteins produced. The proteomic analysis data presented here provide critical informa- tion to compare proteins secreted by Trichoderma reesei and Aspergillus niger when cultivated through submerged and sequential fermenta- tion processes, using steam-explosion sugarcane bagasse as inducer for enzyme production. The proteins were organized according to the families described in CAZy database as cellulases, hemicellulases, proteases/peptidases, cell-wall-protein, lipases, others (catalase, ester- ase, etc.), glycoside hydrolases families, predicted and hypothetical proteins. Further detailed analysis of this data is provided in “Secre- tome analysis …

Hiv Vaccines: Progress, Limitations And A Crispr/Cas9 Vaccine, Omar A. Garcia Martinez

Biology: Student Scholarship & Creative Works

ABSTRACT: The HIV-1 pandemic continues to thrive due to ineffective HIV-1 vaccines. Historically, the world’s most infectious diseases, such as polio and smallpox, have been eradicated or have come close to eradication due to the advent of effective vaccines. Highly active antiretroviral therapy is able to delay the onset of AIDS but can neither rid the body of HIV-1 proviral DNA nor prevent further transmission. A prophylactic vaccine that prevents the various mechanisms HIV-1 has to evade and attack our immune system is needed to end the HIV-1 pandemic. Recent advances in engineered nuclease systems, like the CRISPR/Cas9 system, have …

Genetic Engineering And Sustainable Crop Disease Management: Opportunities For Case-By-Case Decision-Making, Paul Vincelli

Plant Pathology Faculty Publications

Genetic engineering (GE) offers an expanding array of strategies for enhancing disease resistance of crop plants in sustainable ways, including the potential for reduced pesticide usage. Certain GE applications involve transgenesis, in some cases creating a metabolic pathway novel to the GE crop. In other cases, only cisgenessis is employed. In yet other cases, engineered genetic changes can be so minimal as to be indistinguishable from natural mutations. Thus, GE crops vary substantially and should be evaluated for risks, benefits, and social considerations on a case-by-case basis. Deployment of GE traits should be with an eye towards long-term sustainability; several …

Exploring Barriers To The Adoption Of Pharmacogenomic Technology In The Clinical Setting By Clinical Healthcare Providers, Jason Sudia

Seton Hall University Dissertations and Theses (ETDs)

The changing landscape of healthcare in the US has created new questions about how to best provide cost-effective, individualized care. Personalized medicine and more specifically, pharmacogenomic technology have offered new tools for healthcare providers to use to increase the efficacy, safety, and cost-effectiveness of care. However, these tools are not being utilized to their predicted extent in the clinical setting. This study utilized Rogers’ Diffusion of Innovations theory to investigate some of the reasons why. A multi-question survey, the PI-created, Pharmacogenomic Adoption Instrument (PAI) ©, was developed to assess the knowledge, attitudes and experience concerning pharmacogenetic technology in a spectrum …