Biotechnology Commons^™

A Novel Method Of Rapid Detection For Heavy Metal Copper Ion Via A Specific Copper Chelator Bathocuproinedisulfonic Acid Disodium Salt, Yali Wang, Tinglin Ma, Joseph Brake, Zhaoyue Sun, Jiayu Huang, Jing Li, Xiaobin Wu

Department of Biochemistry: Faculty Publications

The extensive usage and production of copper may lead to toxic effects in organisms due to its accumulation in the environment. Traditional methods for copper detection are time consuming and infeasible for field usage. It is necessary to discover a real-time, rapid and economical method for detecting copper to ensure human health and environmental safety. Here we developed a colorimetric paper strip method and optimized spectrum method for rapid detection of copper ion based on the specific copper chelator bathocuproinedisulfonic acid disodium salt (BCS). Both biological assays and chemical methods verified the specificity of BCS for copper. The optimized reaction …

Mass Spectrometry Of The White Adipose Metabolome In A Hibernating Mammal Reveals Seasonal Changes In Alternate Fuels And Carnitine Derivatives, Frazer I. Heinis, Sophie Alvarez, Matthew T. Andrews

Nebraska Center for Biotechnology: Faculty and Staff Publications

Mammalian hibernators undergo substantial changes in metabolic function throughout the seasonal hibernation cycle. We report here the polar metabolomic profile of white adipose tissue isolated from active and hibernating thirteen-lined ground squirrels (Ictidomys tridecemlineatus). Polar compounds in white adipose tissue were extracted from five groups representing different timepoints throughout the seasonal activity-torpor cycle and analyzed using hydrophilic interaction liquid chromatography-mass spectrometry in both the positive and negative ion modes. A total of 224 compounds out of 660 features detected after curation were annotated. Unsupervised clustering using principal component analysis revealed discrete clusters representing the different seasonal timepoints throughout hibernation. One-way …

Putrescine Detected In Strains Of Staphylococcus Aureus, Javier Seravalli, Frank Portugal

Department of Biochemistry: Faculty Publications

Most forms of life, including the archaea, bacteria, and eukaryotes synthesize the polyamine putrescine. Although putrescine is widely distributed, several Gram-positive bacteria, including Staphylococcus aureus (S. aureus), appear to be the exceptions. We report here that strains of S. aureus can produce the polyamine putrescine, as well as the derivative N-acetyl-putrescine. Three strains of S. aureus from the American Type Culture Collection (ATCC), one strain listed in the National Center for Biotechnology Information (NCBI) database, whose genomic sequence is well defined, and well as eight strains from S. aureus-induced brain abscesses of individual patients from multiple …

The Effect Of Acidic Ph On Growth Kinetics, Biomass Productivity, And Prima-Ry Metabolite Contents Of Euglena Sp., Istini Nurafifah, Muhammad Andhi Hardianto, Tia Erfianti, Ria Amelia, Khusnul Qonita Maghfiroh, Dedy Kurnianto, Dwi Umi Siswanti, Brilian Ryan Sadewo, Renata Adaranyssa Egistha Putri, Eko Agus Suyono

Makara Journal of Science

Euglena is a microalga with the potential to be an environmentally friendly renewable energy resource. The pH value is a crucial factor in micro-algal cultivation. Changes in pH affect the growth and development of microalgae, including the production of biomass and primary metabolites, such as proteins, carbohydrates, and lipids. In this study, Euglena sp. was grown on Cramer-Myers medium and subjected to various acidic conditions. This study aimed to determine the effect of pH on the growth kinetics, biomass, carbohydrate, lipid, and protein contents of Euglena sp. The Euglena sp. culture was optimized at various pH values of 2.5, 3.5, …

Enhancing Lovastatin Biosynthesis In Oyster Mushrooms (Pleurotus Ostreatus) Using Phytohormones, Fransisca Astrid Mustafa, Mamat Kandar, I Nyoman Pugeg Aryantha

Makara Journal of Science

The biosynthesis of lovastatin, the anti-cholesterol compound, in oyster mushrooms (Pleurotus ostreatus), has the potential to be enhanced by utilizing phytohormones, which activate the expression of certain genes. This research aimed to determine the best type and concentration of phytohormone among auxin, gibberellin, and cytokinin, as well as the best mycelial colonization percentage in F2 medium to realize the greatest increase in lovastatin biosynthesis in oyster mushrooms. Lovastatin was extracted from the fruiting bodies and mycelia and analyzed by spectrophotometry. The analysis of CYP450 linked to lovastatin biosynthesis was conducted by quantitative polymerase chain reaction (qPCR) using samples containing …

Type Iv Pilus-Mediated Inhibition Of Acinetobacter Baumannii Biofilm Formation By Phenothiazine Compounds, Nam Vo, Benjamin S. Sidner, Yafan Yu, Kurt H. Piepenbrink

Department of Biochemistry: Faculty Publications

Infections by pathogenic Acinetobacter species represent a significant burden on the health care system, despite their relative rarity, due to the difficulty of treating infections through oral antibiotics. Multidrug resistance is commonly observed in clinical Acinetobacter infections and multiple molecular mechanisms have been identified for this resistance, including multidrug efflux pumps, carbapenemase enzymes, and the formation of bacterial biofilm in persistent infections. Phenothiazine compounds have been identified as a potential inhibitor of type IV pilus production in multiple Gram-negative bacterial species. Here, we report the ability of two phenothiazines to inhibit type IV pilus-dependent surface (twitching) motility and biofilm formation …

Cytotoxic Activity Of Salvia Officinalis Extract Mediated By The Inhibition Of Jak2 In Breast Cancer, Salma Alawadi Dawood

Theses and Dissertations

Background: Breast cancer is the most common type of invasive cancer in women in their forties and fifties. Recent evidence suggests that JAK2/STAT3 signaling is constitutively active in breast cancer. Previous studies suggest that plant extracts, including Salvia Officinalis, have strong cytotoxic effects on breast cancer cells. The differential expression of miRNAs is also strongly linked to cancer development.

Aim: In the current study, we hypothesize that S. Officinalis extract suppresses JAK2 expression and has strong anticancer potential in MCF7 breast cancer cell lines.

Methods: GC-MS analysis showed the presence of flavonoids in dried leaf of …

Association Of Gut Microbiota Composition With Alzheimer’S-Like Disease In A Rat Model, Hana Heiba

Theses and Dissertations

With many countries turning ‘grey’ and facing an issue with aging populations, the risk of developing one form of neurodegenerative disease is increasing. Dementia, being the most common syndrome resulting from neurodegeneration, severely affects memory and cognitive functions. Alzheimer’s disease (AD) is the most common neurodegenerative disorder, with an estimated 615,000 new cases will be added to the existing 5.7 million by 2030 (Alkasir et al., 2017).

In the current study, we establish a sporadic AD-like rat model by injecting STZ intracerebrally. Stool samples were collected at two time points; after three weeks for the acute stage, and 3 months …

Measuring The Physiological Effects Of Methylobacterium Oryzae Foliar Inoculation Onto Salt-Stressed Tomatoes Grown In A Greenhouse Semi-Hydroponic System, Collin Dice

Master of Science in Integrative Biology Theses

Salinity stress has been shown to improve hydroponic produce quality such as that of tomato but at the trade-off of produce yield. In order to improve demand for hydroponic produce, techniques that improve salinity tolerance could offset declines in yield without altering the improved quality of produce. In two separate trials, we tested whether foliar inoculation of the ACC deaminase-producing bacteria Methylobacterium oryzae CBMB20 improved salt tolerance in tomatoes grown in a semi-hydroponic system using an indeterminate variety exposed to 26mM NaCl (Trial 1) and a determinate variety exposed to 17mM NaCl (Trial 2). We also tested how salinity and …

Effects Of Interleukin-22 (Il-22) On Necroptosis, Inflammatory Responses, And Metabolism Of Mycobacterium Tuberculosis Stimulated Type 2 Diabetes Mellitus Mouse Macrophages, Bismark Owusu-Afriyie

Biotechnology Theses

Mycobacterium tuberculosis (Mtb) infects one-third of the world’s population and causes nearly 1.3 million deaths per year. Limited information is available about the immune responses during Mtb infection in type 2 diabetic hosts. Our laboratory developed an experimentally induced type 2 diabetes (T2DM) model in wild-type C57BL/6 mice and found that IL-22 and type 3 innate lymphoid cells (ILC3s) reduce inflammation and mortality of Mtb-infected T2DM mice. Our laboratory also found that Mtb-infected alveolar macrophages (AMs) from T2DM mice undergo necroptosis compared to Mtb-infected AMs of non-T2DM mice. In the current study, we determined whether …

Artificial Scaffold And Neural Stem Cell For Neural Regeneration In Rats With Spinal Cord Injury, Krishna Deo Sharma

Student Theses and Dissertations

Spinal cord injury (SCI) is a devastating condition that often leads to permanent loss of sensory and motor function. Unfortunately, current treatments only manage symptoms and do not provide a complete cure. In this study, we investigated the effectiveness of 3D printed scaffolds and nanomaterials (gold nanorods substrate, nanocellulose-lysine, and TEMPO cellulose) in supporting the growth and differentiation of neural stem cells (NSCs) in vitro. We found that these materials were biocompatible, non-toxic, and able to promote the growth and differentiation of NSCs. Additionally, the unique mechanical properties of these nanomaterials make them a promising candidate for use in neuroregeneration …

Hematopoietic Stem Cells And Betaherpesvirus Latency, Lindsey B. Crawford

Department of Biochemistry: Faculty Publications

The human betaherpesviruses including human cytomegalovirus (HCMV), human herpesvirus (HHV)-6a and HHV-6b, and HHV-7 infect and establish latency in CD34+ hematopoietic stem and progenitor cells (HPCs). The diverse repertoire of HPCs in humans and the complex interactions between these viruses and host HPCs regulate the viral lifecycle, including latency. Precise manipulation of host and viral factors contribute to preferential maintenance of the viral genome, increased host cell survival, and specific manipulation of the cellular environment including suppression of neighboring cells and immune control. The dynamic control of these processes by the virus regulate inter- and intra-host signals critical to the …

Compositional Properties Of Flours And Starches From The Philippine National Seed And Industry Council-Registered Root Crops, Edgardo E. Tulin, Chinelo M. Cardaño, Anabella B. Tulin, Ma. Theresa P. Loreto, Ea Kristine Clarisse B. Tulin, Ma. Villa N. Yu

The Philippine Agricultural Scientist

Based in the Visayas State University, Philippines, the germplasm collection of the Philippine Root Crops Research and Training Center (PhilRootcrops) is home to several varieties that have not been fully characterized for their potential use in the production of functional foods. This study evaluated the compositional properties, amylose/amylopectin ratio, and glycemic index of flour and starches of 10 varieties of cassava (Manihot esculenta Crantz), sweet potato (Ipomoea batatas L. (Lam)), and taro (Colocasia esculenta L. Schott); 8 varieties of yam (Dioscorea alata L.); and 5 varieties of arrowroot (Maranta arundinacea L.), which was the recommended …

Development Of Wiskott-Aldrich Syndrome Knock Out Protocol For Drug Substance Assay Development, Julia C. Hanna

Master's Theses

Wiskott-Aldrich Syndrome (WAS) is a rare X-linked primary immunodeficiency affecting approximately 1 in 100,000 live XY births in North America and is caused by a mutation to the WAS gene which is expressed across hematopoietic lineages. The WAS protein (WASp) plays a role in regulating actin polymerization. On a cellular level, there are a variety of effects of a lack of WASp or expression of a dysfunctional WASp protein for patients including issues with migration, adhesion, chemotactic response, phagocytosis, activation, and proliferation across different cell types in addition to reduced platelet size and output. This can lead to several systematic …

Bola3 And Nfu1 Link Mitoribosome Iron-Sulfur Cluster Assembly To Multiple Mitochondrial Dysfunctions Syndrome, Hui Zhong, Alexandre Janer, Oleh Khalimonchuk, Hana Antonicka, Eric Shoubridge, Antoni Barrientos

Department of Biochemistry: Faculty Publications

The human mitochondrial ribosome contains three [2Fe-2S] clusters whose assembly pathway, role, and implications for mitochondrial and metabolic diseases are unknown. Here, structure-function correlation studies show that the clusters play a structural role during mitoribosome assembly. To uncover the assembly pathway, we have examined the effect of silencing the expression of Fe-S cluster biosynthetic and delivery factors on mitoribosome stability. We find that the mitoribosome receives its [2Fe-2S] clusters from the GLRX5-BOLA3 node. Additionally, the assembly of the small subunit depends on the mitoribosome biogenesis factor METTL17, recently reported containing a [4Fe-4S] cluster, which we propose is inserted via the …

The Development Of A Primer Payload With Microparticles For Uti Pathogen Identification Using Polythymidine- Modified Lamp Primers In Droplet Lamp, Jonas Otoo

KGI Theses and Dissertations

Nucleic acid amplification tests (NAATs) are among the diagnostic tests with the highest sensitivity and specificity. However, they are more complex to develop than other diagnostic tests such as biochemical tests and lateral flow immunoassay tests. Polymerase chain reaction (PCR) is the gold standard for NAATs. PCR requires thermal cycling to achieve clonal amplification of the target pathogen DNA for diagnosis. Thermal cycling poses a challenge in the development of PCR diagnostics for point-of-care (POC) settings. Loop-mediated isothermal amplification (LAMP) offers an isothermal method for NAATs diagnostics. The advancement of the microfluidics field significantly enhances the development of LAMP diagnostics …

Dietary Marine-Derived Ingredients For Stimulating Hair Cell Cycle, Aleksandra Augustyniak, Helena Mcmahon

Publications

In normal condition human hair growth occurs through three phases, anagen (growth phase included about 85 % of hairs, last from 2 to 6 years), catagen (transitional phase lasting up to 2 weeks) and telogen (resting phase which last from 1 to 4 months). Natural dynamics of the hair growth process can be impaired by several factors, such as genetic predisposition, hormonal disorders, aging, poor nutrition or stress, which can lead to the slowdown in the growth of hair or even hair loss. The aim of the study was to assess the hair growth promotion effect of marine-derived ingredients, hair …

On The Resurrection Of Microbes: An Eco-Christological Approach To The Resurrection, Denys Janiga Osb

Obsculta

This article attempts to bring ecology and Christology into conversation through a transdisciplinary approach. It looks at the pastoral implications of eco-anxiety, resurrection ecology in the field of biotechnology, Church teaching on the Resurrection, and concludes by initiating an interpretation of the resurrection appearances in the Gospel of Luke through the lens of the microbiome.

Extravasated Brain-Reactive Autoantibodies Perturb Neuronal Surface Protein Expression In Alzheimer's Pathology, Wardah Bajwa, Mary Kosciuk, Randel L. Swanson, Anuradha Krishnan, Venkat Venkataraman, Robert Nagele, Nimish Acharya

Rowan-Virtua Research Day

Background: Increased blood-brain barrier (BBB) permeability is reported in both the neuropathological and in vivo studies in both Alzheimer’s Disease (AD) and age matched cognitively normal, no cognitive impairment (NCI), subjects. Impaired BBB allows various vascular components such as immunoglobulin G (IgG) to extravasate into the brain and specifically bind to various neuronal surface proteins (NSP), also known as brain reactive autoantibodies (BrABs). This interaction is predicted to further enhance deposition of amyloid plaques.

Hypothesis: Interaction between extravasated BrABs and its cognate NSPs lower the expression of that NSPs in AD patients.

Methods: We selected Western blotting technique to study …

Green Biorefinery Systems For The Production Of Climate-Smart Sustainable Products From Grasses, Legumes And Green Crop Residues, James Gaffey, Gaurav Rajuaria, Helena Mcmahon, Rajeev Ravindran, Carmen Dominguez, Morten Ambye Jensen, Macella F. Souza, Erik Meers, Marta Macias Aragonés, Dubravka Skunka, Johan P. M. Sanders

Publications

Grasses, legumes and green plant wastes represent a ubiquitous feedstock for developing a bioeconomy in regions across Europe. These feedstocks are often an important source of ruminant feed, although much remains unused or underutilised. In addition to proteins, these materials are rich in fibres, sugars, minerals and other components that could also be used as inputs for bio-based product development. Green Biorefinery processes and initiatives are being developed to better capitalise on the potential of these feedstocks to produce sustainable food, feed, materials and energy in an integrated way. Such systems may support a more sustainable primary production sector, enable …

Assesment Of Structure, Function, And Microevolutionary Dynamics Of Extrachromosomal Circular Dna In Chinese Hamster Ovary Cells, Dylan Chitwood

All Dissertations

Chinese hamster ovary (CHO) cell lines are among the most popular expression hosts used in biopharmaceutical manufacturing due to relative ease of culture, capacity to perform human-like post-translational modifications, and non-susceptibility to viruses. However, the intrinsic plasticity of the CHO genome can lead to undesired genetic rearrangements, phenotypic shifts, reduced product quality, and early culture termination that prevents continuous biomanufacturing. A characteristic of plastic and unstable genomes that is poorly understood in CHO cells is extrachromosomal circular DNA (eccDNA). EccDNAs are focal amplifications of the genome that reside in the extranuclear space. These plasmid-like entities are structurally complex and are …

Rapid & Sensitive Detection Of Antibiotic Resistance Genes By Utilizing New Diagnostic Probe Tales With 2d-Nanosheet Graphene Oxide, Jihye Kang

Masters Theses & Specialist Projects

As antibiotic resistance is one of the major health concerns in infectious diseases because of reducing the efficacy of antibiotics, rapid and sensitive detection of antibiotic resistance genes is critical for the effective treatment of infectious diseases. Polymerase chain reaction (PCR) is widely used to detect nucleic acids, but it requires additional steps involved in DNA denaturation and subsequent hybridization. A new class of DNA-binding domains called transcriptional activator-like effectors (TALEs) provide a novel scaffold for designing versatile DNA-binding proteins due to their modularity and predictability. Here, we developed a simple, rapid, and sensitive system for detecting antibiotic resistance genes …

Review Of Biomedical Applications Of Cardiovascular Tissue Engineering, Natalie M. Howard

Honors College Theses

Tissue engineering can be defined as processes that aim to generate three-dimensional functional tissues in vitrothat have been favorably altered according to the structural, biochemical, electrophysiological, and biomechanical properties of the desired tissue before implantation into the human body. In relation to cardiac tissues, these properties would include the ability to conduct action potentials, withstand systolic pressure, permit sufficient O₂ and CO₂penetration, sufficient vascularization to supply nutrients for cellular activity, surface topology that enables cellular communication, and more. As heart diseases and instances of myocardial infarction continue to rise worldwide, there is an increasing need for …

Impact Of Lyophilization On Porcine Hemoglobin Properties., Mustafa Almosawi

College of Arts & Sciences Senior Theses

Blood transfusion is the single most often performed lifesaving procedure in hospitals worldwide. Unfortunately, packed red blood cells (RBCs) used for transfusion can only be stored for 42 days at 4 °C before being discarded due to irreversible damage that occurs during storage. Any reduction in available RBCs for an extended period can lead to blood shortages. To increase the shelf-life of RBCs, we investigated freeze-drying (lyophilizing) in the presence of the non-toxic sugar trehalose as a method for long-term preservation. However, the oxidative stress of the lyophilization and storage processes can compromise the functionality of these cells, and the …

Laser Inscribed Graphene Aptasensor For Detection Of Apple Stem Pitting Virus, Yifan Tang

All Theses

Apple stem pitting virus (ASPV) is a latent RNA virus that could cause significant loss to apple production; early-stage field diagnosis could reduce the economic loss caused by ASPV infection. Biosensor is a promising user-friendly detection tool that could transduce biological or chemical interactions into electric signals. Among various types of biosensors, aptasensor, which use aptamers as recognition elements show potential as a high throughput, in-field test. The goal of this thesis is to develop an electrochemical aptasensor testing system for ASPV detection. The quality control study of laser inscribed graphene (LIG) based working electrode of the biosensor was conducted …

Investigation The Plugging Behavior Of Virus Filters, Iman Abdulqader Saab

Graduate Theses and Dissertations

A virus filtration step is integral to the manufacturing of biopharmaceuticals to ensure viral safety. A virus filter is a single-use device that uses a size-based separation process and has a unique structure with minimal defects, so that contaminating virus particles cannot pass through the membrane pores, while therapeutic molecules can. The development of novel antibodies (Abs), including significant increases in product titers, is frequently challenged by virus filter fouling, making a better understanding of the underlying mechanisms essential. This thesis focused investigating of the effect of prefilter types, buffer type and salt content on virus filtration performance. The impact …

Comprehensive Long- And Short- Read Sequencing Approaches To Screen The Mutational Landscape After Therapeutic Crispr-Editing For Duchenne Muscular Dystrophy, Mary Jia, Made Harumi Padmaswari, Landon Burcham, Shilpi Agrawal, Alexis Ivy, Abbey Bryan, Christopher E. Nelson

Biomedical Engineering Undergraduate Honors Theses

Duchenne muscular dystrophy (DMD) is an X-linked muscle wasting disease caused by mutations in the Dmd gene resulting in non-functional dystrophin. Adeno-associated virus (AAV)-mediated delivery of classical CRISPR systems resulting in double-stranded breaks (DSBs) has yielded promise as a curative DMD therapeutic. However, the consequences of AAV-CRISPR require more thorough characterization. PCR-enriched short-read amplicon sequencing at cut sites is the most common method of sequencing CRISPR editing, but PCR bias results in unreliable quantification of on-target edits. We analyze orthogonal long- and short-read sequencing approaches to screen the outcomes of CRISPR technologies at the DNA and RNA-level for DMD. We …

Exploration Of Spider Silk Assembly, Natalie Naseib, Joshua Navarro, Nadia Duong, Seohyun Lee, Daniel Kim, Danbir Somal, Sevak Manucharyan, Josephine Kim, Seha Park, Alice Lan, Karen Kwong, Mason Tri

Pacific Undergraduate Research and Creativity Conference (PURCC)

No abstract provided.

Study Of The Assembly Trigger For Spider Silk Proteins, Katina Zhu, Amanda Lee, Megan Lau, Sungkyung Lee, Kaden Chen, Katherine Yoo, David Eum, Daniel Eum, Austin Cho, Jiho Choi

Pacific Undergraduate Research and Creativity Conference (PURCC)

No abstract provided.

Electrochemical Reduction Of Carbon Dioxide Using Ruthenium And Amines, Sydney Moise

Honors Theses

With an increased concern for climate change in the recent years, a significant area of research has been devoted to the reduction of greenhouse gases. Carbon dioxide (CO2) resides in the atmosphere between 300 – 1000 years, making the reduction of the molecule a substantial field of study.1 Amines have been used as CO2 scrubbing agents in literature historically, due to their ability to form bonds to carbon.2 Although studies involving metal catalysts and amines have been reported numerous times, research involving chemical reduction of CO2 using purely amines is scarce. In this paper, amines, in addition to hydride donors …