Microbiology Commons^™

Overcoming Obstacles In Protein Expression In The Yeast Pichia Pastoris: Interviews Of Leaders In The Pichia Field, Zoe Ingram, Abha Patkar, Dahoon Oh, Kevin K. Zhang, Christina Chung, Joan Lin-Cereghino, Geoff P. Lin-Cereghino

Pacific Journal of Health

The yeast Pichia pastoris (also known as Komagataella pastoris) has been used for over 30 years to produce thousands of valuable, heterologous proteins, such as insulin to treat diabetes and antibodies to prevent migraine headaches. Despite its success, there are some common, stubborn problems encountered by research scientists when they try to use the yeast to produce their recombinant proteins. In order to provide those working in this field with strategies to overcome these common obstacles, nine experts in P. pastoris protein expression field were interviewed to create a written review and video (https://www.youtube.com/watch?v=FyC9xXVLyD0). This review describes …

The Role Of Interspecies Recombination In The Evolution Of Antibiotic-Resistant Pneumococci, Joshua C. D'Aeth, Mark Pg Van Der Linden, Lesley Mcgee, Herminia De Lencastre, Paul Turner, Jae-Hoon Song, Stephanie W. Lo, Rebecca A. Gladstone, Gps Consortium, Sadia Shakoor

Department of Pathology and Laboratory Medicine

Multidrug-resistant Streptococcus pneumoniae emerge through the modification of core genome loci by interspecies homologous recombinations, and acquisition of gene cassettes. Both occurred in the otherwise contrasting histories of the antibiotic-resistant S. pneumoniae lineages PMEN3 and PMEN9. A single PMEN3 clade spread globally, evading vaccine-induced immunity through frequent serotype switching, whereas locally circulating PMEN9 clades independently gained resistance. Both lineages repeatedly integrated Tn916-type and Tn1207.1-type elements, conferring tetracycline and macrolide resistance, respectively, through homologous recombination importing sequences originating in other species. A species-wide dataset found over 100 instances of such interspecific acquisitions of resistance cassettes and flanking …

Novel Scalable And Simplified System To Generate Microglia-Containing Cerebral Organoids From Human Induced Pluripotent Stem Cells, Brittany Bodnar, Yongang Zhang, Jinbiao Liu, Yuan Lin, Peng Wang, Zhengyu Wei, Sami Saribas, Yuanjun Zhu, Fang Li, Xu Wang, Wenli Yang, Qingsheng Li, Wen-Zhe Ho, Wenhui Hu

Nebraska Center for Virology: Faculty Publications

Human cerebral organoid (CO) is a three-dimensional (3D) cell culture system that recapitulates the developing human brain. While CO has proved an invaluable tool for studying neurological disorders in a more clinically relevant matter, there have still been several shortcomings including CO variability and reproducibility as well as lack of or underrepresentation of certain cell types typically found in the brain. As the technology to generate COs has continued to improve, more efficient and streamlined protocols have addressed some of these issues. Here we present a novel scalable and simplified system to generate microglia-containing CO (MCO). We characterize the cell …

Decomposition Of Microbial Necromass Is Divergent At The Individual Taxonomic Level In Soil, Weiling Dong, Alin Song, Huaqun Yin, Xueduan Liu, Jianwei Li, Fenliang Fan

Agricultural and Environmental Sciences Faculty Research

The turnover of microbial biomass plays an important part in providing a significant source of carbon (C) to soil organic C. However, whether the decomposition of microbial necromass (non-living microbial biomass) in the soil varies at the individual taxa level remains largely unknown. To fill up these gaps, we compared the necromass decomposition of bacterial and archaeal taxa by separating live microbial biomass with 18O-stable isotope probing from dead microbial biomass in soil. Our results showed that most of the microbial necromass at the operational taxonomic unit level (88.51%), which mainly belong to Acidobacteria, Actinobacteria, Gemmatimonadetes, and Proteobacteria, decomposed significantly …

College Of Natural Sciences Newsletter, July 2021, College Of Natural Sciences

College of Natural Sciences Newsletters and Reports

Volume 2, Issue 7.

Page 1 Dean's Message
Page 2 Celebrating Three Years
Page 3 Summer School Travel
Page 4 Media Coverage of CNS
Page 5 Chemistry Teachers Visit
Page 5 Remembering Dr. Warren Hein
Page 6 Research Spotlights: Undergraduate Summer Experiences
Page 7 Open PRAIRIE Data; Grants Awarded in CNS

Directed Genome Evolution To Identify Genes For Macrophage Survival By Staphylococcus Agnetis, Sonali Lakshika Anne Lenaduwe Lokuge

Graduate Theses and Dissertations

Bacterial Chondronecrosis with Osteomyelitis (BCO) is a debilitating infection that negatively impacts animal welfare and costs the broiler industry billions of dollars annually. We have previously isolated Staphylococcus agnetis 908 from BCO samples obtained from broilers at the University of Arkansas research farm. This isolate can induce BCO lameness at greater than 50% in broilers exposed to the pathogen in drinking water. We found that S. agnetis 908 is capable of surviving and escaping macrophages compared to a closely related cattle isolate,1379. Through Directed Genome Evolution (DGE) we identified that this difference is at least partially associated with an alanine …

Protective Role Of Palmitoleate Against Zika Virus-Induced Apoptosis In Placental Trophoblasts, Philma Glora Muthuraj

College of Education and Human Sciences: Dissertations, Theses, and Student Research

Zika virus (ZIKV) infection in pregnant women causes congenital Zika syndrome which involves birth defects such as intrauterine growth restriction (IUGR), retinal defects and microcephaly in the fetus. ZIKV infection results in placental pathology which plays a crucial role in disease transmission from mother to fetus. Currently, there is no Food and Drug Administration (FDA) approved vaccine or therapeutic drug against ZIKV. In this thesis, we have elucidated the cell signaling pathways connected to ZIKV-induced apoptosis in trophoblasts and also the protective effect of palmitoleate against ZIKV-induced apoptosis. First, we demonstrated the molecular mechanism behind ZIKV-induced apoptosis using an in …

A Yeast Expressed Rbd-Based Sars-Cov-2 Vaccine Formulated With 3m-052-Alum Adjuvant Promotes Protective Efficacy In Non-Human Primates, Maria Pino, Talha Abid, Susan Pereira Ribeiro, Venkata Viswanadh Edara, Katharine Floyd, Justin C. Smith, Muhammad Bilal Latif, Gabriela Pacheco-Sanchez, Debashis Dutta, Shelly Wang, Sanjeev Gumber, Shannon Kirejczyk, Joyce Cohen, Rachelle L. Stammen, Sherrie M. Jean, Jennifer S. Wood, Fawn Connor-Stroud, Jeroen Pollet, Wen Hsiang Chen, Junfei Wei, Bin Zhan, Jungsoon Lee, Zhuyun Liu, Ulrich Strych, Neeta Shenvi, Kirk Easley, Daniela Weiskopf, Alesandro Sette

School of Graduate Studies Faculty Publications

Ongoing SARS-CoV-2 vaccine development is focused on identifying stable, cost-effective, and accessible candidates for global use, specifically in low and middle-income countries. Here, we report the efficacy of a rapidly scalable, novel yeast expressed SARS-CoV-2 specific receptor-binding domain (RBD) based vaccine in rhesus macaques. We formulated the RBD immunogen in alum, a licensed and an emerging alum adsorbed TLR-7/8 targeted, 3M-052-alum adjuvants. The RBD+3M-052-alum adjuvanted vaccine promoted better RBD binding and effector antibodies, higher CoV-2 neutralizing antibodies, improved Th1 biased CD4+T cell reactions, and increased CD8+ T cell responses when compared to the alum-alone adjuvanted vaccine. RBD+3M-052-alum induced a significant …

Bispecific Anti-Hiv Immunoadhesins That Bind Gp120 And Gp41 Have Broad And Potent Hiv-Neutralizing Activity, Seth H. Pincus, Ryan B. Craig, Lauren Weachter, Celia C. Labranche, Rafiq Nabi, Connie Watt, Mark Raymond, Tami Peters, Kejing Song, Grace A. Maresh, David C. Montefiori, Pamela A. Kozlowski

School of Graduate Studies Faculty Publications

We have constructed bispecific immunoglobulin-like immunoadhesins that bind to both the HIV-envelope glycoproteins: gp120 and gp41. These immunoadhesins have N terminal domains of human CD4 engrafted onto the N-terminus of the heavy chain of human anti-gp41 mAb 7B2. Binding of these constructs to recombinant Env and their antiviral activities were compared to that of the parental mAbs and CD4, as well as to control mAbs. The CD4/7B2 constructs bind to both gp41 and gp140, as well as to native Env expressed on the surface of infected cells. These constructs deliver cytotoxic immunoconjugates to HIV-infected cells, but not as well as …

Development Of Vaccines And Antivirals Against Zika Virus, Aryamav Pattnaik

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

The re-emergence of Zika virus (ZIKV), an arbovirus, poses a major global human health concern because of its ability to cause congenital abnormalities and neurological diseases. While many candidate vaccines and antiviral drugs are in the developmental pipeline, none have been approved for use against ZIKV infection. This dissertation describes the characterization of one vaccine and two antiviral drug candidates against ZIKV infection. A bacterial ferritin-based nanoparticle vaccine, termed zDIII-F, is designed to display multiple copies of ZIKV E protein domain III on its surface. These stable nanoparticles are shown to induce robust antibody-mediated protection against lethal ZIKV infection in …

Evaluation Of Horizontal Gene Transfer Between Genetically Engineered Cyanobacteria And Gram-Negative Bacteria, Andriana Chrysovalanti Zourou

Chemistry & Biochemistry Theses & Dissertations

As the world population is increasing and societies become more technology driven, there is an imperative to develop ‘green energy’ sources to protect our planet. Cyanobacteria that have been genetically engineered to produce organic compounds that may be burnt as fuels show great potential, as they are an environmentally friendly and self-renewable, net carbon-neutral option. However, there are potential risks in the development and use of genetically modified organisms (GMOs). We need to understand in advance the risks that GMOs may pose to our environment and to animal and human health. This will enable experimental procedures, containment strategies and policies …

The Persistence Of Human Norovirus Surrogate In Leafy Greens Production System, Wenjun Deng

Graduate Theses and Dissertations

Since mostly consumed raw, foodborne pathogen contamination of leafy greens has led to a large number of foodborne disease outbreaks and illnesses each year in the United States. Human noroviruses (hNoV) are the most common viral pathogen transmitted by leafy greens. In this dissertation, the persistence of the hNoV surrogate Tulane virus (TV) on pre-harvest lettuce and microgreens was investigated. Lettuces are the most studied leafy green model, while previous hNoV research has mainly focused on the post-harvest stage of production. Here, pre-harvest hydroponically grown lettuce were used to determine TV persistence on leafy greens. After inoculation on leaves at …

Functional And Phenotypic Evaluation Of Murine Cytomegalovirus Infection Of The Heart, Cassandra Marie Bonavita

LSU Doctoral Dissertations

Human Cytomegalovirus (HCMV) is a ubiquitous pathogen which infects 50-90% of the world population and results in lifelong latent infection with periodic transient reactivations. In immunocompetent populations acute infection is generally asymptomatic but seropositivity is associated with many chronic diseases, including cardiovascular disease. Despite 139 years of research into HCMV pathogenesis there is still no licensed vaccine and many gaps in the general knowledge of the virus. This includes the role of long term infection as it pertains to chronic diseases. In these studies, murine cytomegalovirus (MCMV) was used as a model of HCMV pathogenesis in the heart. MCMV shares …

Lead And Cadmium Detoxification By Halophilic Bacteria Isolated From Solar Salterns In Lebanon, Nadine G. Zmorrod N. G. Zmorrod, Nawal Al Hakawati, Ranya A. Amer, Hoda H. Yusef

BAU Journal - Science and Technology

Water contamination by heavy metals has gained considerable attention globally. These inorganic pollutants can enter the aquatic environments via different routes thereby threatening biodiversity and human health. Lead and cadmium are hazardous pollutants where their removal by traditional techniques is identified to be costly and ineffective. However, bioremediation by extremophilic microorganisms is considered to be a promising technique as they have considerable potentials to grow in harsh environmental conditions. The present study deals with the isolation of halophilic bacterial isolates from solar salterns in North Lebanon. The isolate H1S9 was identified as Halomonas venusta H9 after showing tolerance to 500 …

Transcriptomic And Functional Investigation Of Bacterial Biofilm Formation, Brooke R. Nemec

USF Tampa Graduate Theses and Dissertations

Staphylococcus aureus and Acinetobacter baumannii are two highly successful human pathogens, which have adopted very different, but effective survival strategies. The success of S. aureus is attributed to the tight regulation of an arsenal of virulence factors. Conversely, A. baumannii lacks what would be considered traditional virulence factors and, instead, has developed a high tolerance for environmental stress, which allows it to persist in unforgiving environments, including nosocomial settings and the human body. One common characteristic of these two organisms is their proclivity for biofilm formation. Herein, we discuss the diverse mechanisms governing biofilm formation for A. baumannii and S. …

Telomeric And Sub-Telomeric Structure And Implications In Fungal Opportunistic Pathogens, Raffaella Diotti, Michelle Esposito, Chang Hui Shen

Publications and Research

Telomeres are long non-coding regions found at the ends of eukaryotic linear chromosomes. Although they have traditionally been associated with the protection of linear DNA ends to avoid gene losses during each round of DNA replication, recent studies have demonstrated that the role of these sequences and their adjacent regions go beyond just protecting chromosomal ends. Regions nearby to telomeric sequences have now been identified as having increased variability in the form of duplications and rearrangements that result in new functional abilities and biodiversity. Furthermore, unique fungal telomeric and chromatin structures have now extended clinical capabilities and understanding of pathogenicity …

Exploring Prokaryotic And Viral Communities Within Florida’S Freshwater Springs, Kema Saydoon Malki

USF Tampa Graduate Theses and Dissertations

Florida’s freshwater springs serve as a crucial ecosystem for the state, having both ecological and economic importance. Springs occur where there is an opening where aquifers discharge groundwater. Florida’s springs are fed by the Floridan aquifer system (FAS), which underlies the entire state and extends into several neighboring states. The FAS provides potable water to over 10 million people. The constant flow of groundwater through the springs feeds major river systems and creates unique freshwater systems that support numerous plants and animals. The combination of natural beauty and charismatic fauna attract tourism and generate millions of dollars in revenue for …

Host Phylogeny And Diet Shape Gut Microbial Communities Within Bamboo-Feeding Insects, Kuanguan Huang, Jie Wang, Junhao Huang, Shouke Zhang, Alfried P. Vogler, Quanquan Liu, Yongchun Li, Maowei Yang, You Li, Xuguo Zhou

Entomology Faculty Publications

The gut microbiome plays an important role in a host’s development and adaption to its dietary niche. In this study, a group of bamboo-feeding insects are used to explore the potential role of the gut microbiota in the convergent adaptation to extreme diet specialization. Specifically, using a 16S rRNA marker and an Illumina sequencing platform, we profiled the microbial communities of 76 gut samples collected from nine bamboo-feeding insects, including both hemimetabolous (Orthoptera and Hemiptera) and holometabolous (Coleoptera and Lepidoptera) species, which are specialized in three distinct dietary niches: bamboo leaf, shoot, and sap. The gut microbiota of these insects …

A Bacterial Inflammation Sensor Regulates C-Di-Gmp Signaling, Adhesion, And Biofilm Formation, Arden Perkins, Dan A. Tudorica, Raphael D. Teixeira, Tilman Schirmer, Lindsay Zumwalt, O. Maduka Ogba, C. Keith Cassidy, Phillip J. Stansfeld, Karen Guillemin

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Bacteria that colonize animals must overcome, or coexist, with the reactive oxygen species products of inflammation, a front-line defense of innate immunity. Among these is the neutrophilic oxidant bleach, hypochlorous acid (HOCl), a potent antimicrobial that plays a primary role in killing bacteria through nonspecific oxidation of proteins, lipids, and DNA. Here, we report that in response to increasing HOCl levels, Escherichia coli regulates biofilm production via activation of the diguanylate cyclase DgcZ. We identify the mechanism of DgcZ sensing of HOCl to be direct oxidation of its regulatory chemoreceptor zinc-binding (CZB) domain. Dissection of CZB signal transduction reveals that …

Oral Vaccination Approaches For Anti-Shiv Immunity, Erandi Velarde De La Cruz, Lingyun Wang, Deepanwita Bose, Sailaja Gangadhara, Robert L. Wilson, Rama R. Amara, Pamela A. Kozlowski, Anna Aldovini

School of Graduate Studies Faculty Publications

We modified a Sabin Oral Poliovirus Vaccine (OPV) vector to permit secretion of the antigens of interest with the goal of improving anti-HIV Env humoral responses in a SHIV mucosal immunization composed of DNA and recombinant OPVs. We evaluated stimulation of systemic and mucosal cell-mediated and humoral immunity in Rhesus macaques by two regimens, both involving a prime with a SHIVBG505 DNA construct producing non-infectious particles formulated in lipid nanoparticles, administered in the oral cavity, and two different viral vector boostings, administered in the oral cavity and intestinally. Group 1 was boosted with rMVA-SHIVBG505, expressing SIV Gag/Pol and HIVBG505 Env. …