Laboratory and Basic Science Research Commons^™

Surveillance Of Wisconsin Organisms For Trends In Antimicrobial Resistance And Epidemiology (Swotare): 2018-2019 Report On Enterobacter Cloacae And Klebsiella Pneumoniae Clinical Isolates, Alyssa Reynoso, Erik Munson

Clinical Lab Sciences Faculty Research and Publications

Objectives: Both Enterobacter cloacae and Klebsiella pneumoniae have been regarded as important opportunistic pathogens for humans. Recent data have described the spread of multi-resistant strains of these organisms. Development of novel resistance phenotypes may result in a reduction in anti-infective efficacy, therefore making patient treatment decisions challenging. The Surveillance of Wisconsin Organisms for Trends in Antimicrobial Resistance and Epidemiology (SWOTARE) program aims to combat this issue and improve antibiotic stewardship by monitoring antimicrobial resistance at a local level.

Design: Multi-center laboratory surveillance, with testing at a single location utilizing standardized media and susceptibility testing protocols

Methods: In the years 2018 …

The Relationship Between Climate And Nesting Patterns In Two-Lined Salamanders Spanning The Northeast To Southeast United States, Jasmyne Blake-Sinclair

Symposium of Student Scholars

Two-lined salamanders (Eurycea bislineata species complex) are among the most ubiquitous amphibians in the Eastern United States. Because this group is geographically widespread and shows notable life history variation, it provides an opportunity to study the role of phylogeny and environment in shaping these differences. However, records of some important life history traits—such as clutch size and the date of oviposition—are scarce in the scientific literature. In this study, we gathered observations from published research, field notes, and citizen science databases like iNaturalist—a site where anyone can share natural history photographs and pinned geographical locations for other users to …

Blood–Brain Barrier Breakdown And Astrocyte Reactivity Evident In The Absence Of Behavioral Changes After Repeated Traumatic Brain Injury, Celeste Dunn, Nasya Sturdivant, Sara Venier, Syed Ali, Jeffery Wolchok, Kartik Balachandran

Biomedical Engineering Faculty Publications and Presentations

Repeated traumatic brain injuries (TBIs) cause debilitating effects. Without understanding the acute effects of repeated TBIs, treatment options to halt further degeneration and damage cannot be developed. This study sought to examine the acute effects of blood–brain barrier (BBB) dysfunction, edema, inflammation and behavioral changes after either a single or double TBI using a C57BL/6 mouse model. We examined the effects of one or two TBIs, of either a mild or moderate severity. Double injuries were spaced 7 days apart, and all analysis was performed 24 h post-injury. To examine edema and inflammation, protein levels of glial fibrillary acidic protein …

Simultaneous Determination Of Avanafil And Dapoxetine In Human Plasma Using Liquid Chromatography/Tandem Mass Spectrometry (Lc-Ms/Ms) Based On A Protein Precipitation Technique, Ekram Hany Mohamed, Mohamed S. S. Attia, Mona N. Abou-Omar,, Abdelaziz M Annadi,, Ahmed O. Youssef, Mohamed A. Amin, Noha M. El Zahar,

Pharmacy

A rapid and selective LC-MS/MS method is described for the simultaneous assay of Avanafil and Dapoxetine in human plasmaviaa protein precipitation (PP) sample preparation technique. Tadalafil was chosen as the internal standard reaching good recovery and reproducibility while diminishing the effects of the matrix. An Agilent Zorbax Eclipse XDB C18column (4.6 × 50 mm, 1.8 μm) was used for the chromatographic separation and analysis, while 0.1% formic acid : acetonitrile (60 : 40, v/v) was utilized at a flow rate of 0.5 mL min−1. It was revealed that 6 min stop time accomplished the best separation. The assay was linear …

Novel Optical Biosensor Based On A Nano-Gold Coated By Schiff Base Doped In Sol/Gel Matrix For Sensitive Screening Of Oncomarker Ca-125, Ekram Hany Mohamed, Mohamed S. S. Attia, Mona N. Abou-Omar,, Mohammed A. Amin,, Rabah Boukherroub, Hisham Afify

Pharmacy

The urge for sensitive, facile, minimally invasive, and fast detection method of CA-125, a significant and crucial biomarker in ovarian malignancy, is currently substantial. This paper describes the detailed construction and characterization of a newly designed optical nano-biosensor to detect CA-125 accurately and sensitively. The fabricated sensor consists of a nano-gold thin film doped into a matrix of sol-gel, exhibiting a centered fluorescence band at 423 nm when excited at 340 nm. The quantification of CA-125 relies on its quenching ability of this fluorescence signal. The sensor was challenged to evaluate its sensitivity and specificity in detecting CA-125 present in …

Investigations In The Cellular And Molecular Biology Of Human Airway Mucociliary Tissue, Vincent Manna

Graduate School of Biomedical Sciences Theses and Dissertations

Our laboratory has integrated the use of a human-derived, in vitro model of airway mucociliary tissue. We isolate human nasal epithelial cells (HNECs) from the nasal mucociliary tissue of donors with a small brush and expand the airway progenitor cells in culture. The HNECs are then seeded onto semi-permeable transwell inserts. The inserts are in contact with the media in the lower chamber but don’t contain media in the upper chamber therefore the cells are exposed to the air while drawing nutrients from the media below, this is called the Air-Liquid Interface (ALI). HNECs cultured at the ALI initiate a …

Effects Of Trans-Acting Factors On The Translational Machinery In Yeast, Brandon M. Trainor

Graduate School of Biomedical Sciences Theses and Dissertations

Synthesis of proteins, or translation, is a complex biological process requiring the coordinated effort of numerous protein and RNA factors. Central to translation is the ribosome, a complex macromolecular complex consisting of both ribosomal RNA (rRNA) and ribosomal protein (r-protein). Ribosomes are essential and are one of the oldest and most abundant biomolecules across all forms of life. In addition to the ribosome, translation requires messenger RNA (mRNA), transfer-RNA conjugated to an amino acid (aa-tRNA), translation factors, and energy in the form of ATP and GTP. Translation universally occurs in four major stages, initiation, elongation, termination, and recycling, with initiation …

Modulation Of Glucose Homeostasis By Nucleotide P2y2 Receptor And Biological Sex, Hailee Anne Marino

Graduate Theses/Dissertations

Recent insights into the pathological role of Nucleotide P2Y₂ receptor suggest P2Y₂R involvement in high fat diet-induced obesity and potentiates insulin resistance. However, these recent insights do not demonstrate how P2Y₂R modulates glucose homeostasis under physiological conditions. Further, it remains unknown how sex biological factors influence P2Y₂R receptor signaling in the regulation of glucose homeostasis. The research objective for the present study is to elucidate the novel roles of P2Y₂ in fasting blood glucose and glucose tolerance (basal insulin sensitivity) under resting conditions in males and females. We expected that under physiological …

Unveiling Global Roles Of G-Quadruplexes And G4-22 In Human Genetics, Ruth Barros De Paula

Dissertations and Theses (Open Access)

G-quadruplexes are non-B DNA structures formed by four or more runs of repeated guanines that confer unique features to living organism’s genomes. These sequences are enriched in regulatory regions, such as promoters and 5’ UTRs, and have distinct regulatory roles in both health and disease states. Even though previous studies showed the impact of G4 in gene expression, none of them summarized the location-specific effect of G4. Also, there is no broad understanding about the most common G4 repeat in the human genome, named here as G4-22, and how it links to the evolution of mammals and their biology. In …

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 …

Late Embryogenesis Abundant Proteins And Mitochondrial Membranes From An Animal Extremophile: Insights Into Severe Water Stress, John Marcus Anderson

LSU Doctoral Dissertations

This dissertation focuses on two major topics germane to mechanisms by which animals tolerate extreme water stress. First, the impact of transgenic expression of late embryogenesis abundant proteins (LEA) on water stress tolerance of the fruit fly Drosophila melanogaster was investigated. The overall aim was to extend current understanding of the protective properties of LEA proteins documented with isolated cells to a desiccation-sensitive organism during exposure to drying and hyperosmotic stress. Four lines of D. melanogaster were created that expressed transgenes encoding selected LEA proteins originally identified in embryos of the extremophile Artemia franciscana. After 80% tissue water loss, …

Comparative Genomics Methods And Applications, Emily N. Alden

Biomedical Sciences ETDs

Virtually all fields of biology have benefited from the advancements in comparative genomics technologies, specifically in the study of evolution. In this dissertation I develop and use comparative genomic technologies to investigate the novel SARS-CoV-2 virus, assembly the first genome of the black lace domestic angelfish and identify germline genetic variants associated with altered breast cancer-specific survival. Our genome tiling array for the novel coronavirus presents a rapid and cost-effective method to sequence the entire viral genome and can be used to track the rapid evolution of viral variants in the population. The domestic angelfish is a member of the …

All Solid-State Miniaturized Potentiometric Sensors For Flunitrazepam Determination In Beverages, Ekram Hany Mohamed, Mahmoud Tantawy, Ali Yahia

Pharmacy

Flunitrazepam is one of the frequently used hypnotic drugs to incapacitate victims for sexual assault. Appropriate diagnostic tools should be available to victims regarding the growing concern about “date-rape drugs” and their adverse impact on society. Miniaturized screen-printed potentiometric sensors offer crucial point-of-care devices that alleviate this serious problem. In this study, all solid-state screen-printed potentiometric flunitrazepam sensors have been designed. The paper device was printed with silver and carbon ink. Formation of an aqueous layer in the interface between carbon-conducting material and ion-sensing membrane nevertheless poses low reproducibility in the solid-contact electrodes. Accordingly, poly(3,4-ethylenedioxythiophene) (PEDT) nano-dispersion was applied as …

Biographical Feature: Karen C. Carroll, M.D., Erik Munson

Clinical Lab Sciences Faculty Research and Publications

No abstract provided.

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 …

Validation Of A Novel Uplc-Ms/Ms Method For Estimation Of Metformin And Empagliflozin Simultaneously In Human Plasma Using Freezing Lipid Precipitation Approach And Its Application To Pharmacokinetic Study, Ekram Hany Mohamed, Mona N. Abou-Omar,, Mohamed S. S. Attia, Ahmed O. Youssef, Sarah Alharthi,, M. Kenawy

Pharmacy

fast, sensitive one step UPLC ESI-MS/MS method was successfully applied for the simultaneous estimation of two concurrently administrated antidiabetic drugs, Metformin (MET) and Empagliflozin (EMPA) in human plasma. Metformin-d6 (MET-d6) and Empagliflozin-d4 (EMPA-d4) were utilized as internal standards. Extraction of the analytes from the human plasma was performed through acetonitrile precipitation technique followed by freezing the precipitated plasma proteins and lipids to minimize the matrix effect. Chromatographic analysis was developed on Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 50 mm) using isocratic elution mode. A mobile phase of formic acid (0.01 %): acetonitrile (70:30 v/v) with a flow …

Expression And Purification Of Proteins For A Structural Determination: Orf8, Paivy1, Paivy2, Sarah Fashinasi, Avery Moss

Symposium of Student Scholars

Protein purification is a crucial procedure in order to analyze the behavior, structure, and function of a select protein. The Orf8 protein is an increasingly mutable accessory protein found within the SARS-CoV2 virus, however the complex and unpredictable nature of this protein has been met by its inability to be purified thus far. By purifying the model proteins, PaIVY1 and PaIVY2, we believe that we can denote a sound purification method to apply to the Orf8 protein. In our procedure, we transformed BL21(DE3)pLysS cells and inoculated them in LB media. The cells were then placed into a centrifuge to isolate …

The Probability Of Success-Through Fault Tree Analytical Binomial Result, Tyleka Riddle

Night Flight: Online Literary Journal

Prestigious Appeal Rule of Thumb (Ear Drum Plugs)

In truant fashion, with proven accidental probabilistic perception for proposed cued facial expression recognition and voice commands without filtering. The estimated cost analysis for design and development generated from the enhanced project in the word "no”: Additionally, an audio noise effect size (ES) for desired level of comprehensive sensory receptors in sound reference to noisy party syndrome. Sytoss, (2019) "Resulting in a cocktail party theory for chalkboard distinction and conversation with applied facial recognition. As well as, the enhanced project management tool, EPMT, to provide output and input determination for higher-level acoustic …

Developing An Inquiry-Based Laboratory Project For Chem 142l Course At Bsu, Manuel Pina

Honors Program Theses and Projects

In addition to content knowledge, critical and independent thinking, scientific reasoning, and problem-solving skills are essential in preparing next generation of successful workforce. Since one of the biggest advantages of STEM disciplines is a “must-have” hands-on laboratory experience, it is intuitive to exploit this learning space to reinforce afore-mentioned skills. In this context, project-based (PBL) or inquiry-based (IBL) laboratory experiences are rapidly becoming mainstream pedagogical choice for many STEM instructors across United States.^[1-4] PBL, and IBL are learning experiences that offer students an opportunity to experience realistic scientific process of discovery through carefully designed inquiry-driven and/or open-ended investigative laboratory …

Search For Palladin, An Actin-Associated Protein, In Pig Retinal Pigmented Epithelium And Its Role In Epithelial-Mesenchymal Transition, Katrina Powell

Undergraduate Theses

This study investigates the expression of Palladin, a phosphoprotein product of the PALLD gene, in the retinal pigmented epithelium (RPE). Palladin is an actin cross-linking protein and plays a role in cell adhesion and motility. Published reports have demonstrated that a down regulation of Palladin in colon cancer cells results in a reorganization of the actin cytoskeleton, causing the cells to lose their typical shape, become proliferative and migratory. This process is otherwise known as epithelial-mesenchymal transition (EMT). A similar EMT phenomenon is observed when the RPE is exposed to the vitreous humor in patients with proliferative vitreoretinopathy (PVR). In …