Biotechnology Commons^™

Investigating The Potential Of A Cell-Based Gene Editing Therapy For Inherited Metabolic Liver Disease, Ilayda Ates

All Dissertations

Inherited metabolic diseases (IMDs) affecting the liver are relatively rare but collectively have a prevalence of 1 in 800 live births. These diseases result from autosomal recessive single-gene mutations, leading to organ dysfunction and potentially fatal consequences if left untreated. One potential therapeutic strategy for IMDs of the liver involves using CRISPR-Cas9-induced loss of function mutations. However, translating this approach into the clinic is limited by the need for safe and effective CRISPR delivery methods. Adeno-associated viral vectors (AAVs), commonly used for CRISPR delivery, are associated with significant safety and efficacy concerns, including risks for immunogenicity, off-target mutagenesis, and genotoxicity …

Mirna Regulation Of Skeletal Muscle Hypertrophy, Maslyn Greene

All Dissertations

Muscle tissue is vital to animal survival and economically essential to livestock producers. Reduced muscle growth can occur from various reasons, including genetics, nutrition, and the pre and postnatal environment. Understanding the mechanisms behind muscle growth can aid in developing strategies to increase muscle mass. Several methods to increase muscle mass in livestock animals are currently being pursued, including nutrition, hormonal therapy, and genetic selection. One newly developing avenue is using miRNA technologies to change the expression of protein-coding genes. miRNA are a type of RNA that regulate over half of protein-coding gene expression. Here we evaluate muscle characteristics such …

Portable Optical Spectroscopic Assay For Non-Destructive Measurement Of Key Metabolic Parameters On In Vitro Cancer Cells And Organotypic Fresh Tumor Slices, Jing Yan, Carlos Frederico Lima Goncalves, Madison O. Korfhage, Zahid Hasan, Teresa Whei-Mei Fan, Xiaoqin Wang, Caigang Zhu

Markey Cancer Center Faculty Publications

To enable non-destructive metabolic characterizations on in vitro cancer cells and organotypic tumor models for therapeutic studies in an easy-to-access way, we report a highly portable optical spectroscopic assay for simultaneous measurement of glucose uptake and mitochondrial function on various cancer models with high sensitivity. Well-established breast cancer cell lines (MCF-7 and MDA-MB-231) were used to validate the optical spectroscopic assay for metabolic characterizations, while fresh tumor samples harvested from both animals and human cancer patients were used to test the feasibility of our optical metabolic assay for non-destructive measurement of key metabolic parameters on organotypic tumor slices. Our optical …

N-Acetyltransferase 9 Ameliorates Aβ42-Mediated Neurodegeneration In The Drosophila Eye, Prajakta Deshpande, Anuradha Venkatakrishnan Chimata, Emily E. Snider, Aditi Singh, Madhuri Kango-Singh, Amit Singh

Biology Faculty Publications

Alzheimer’s disease (AD), a progressive neurodegenerative disorder, manifests as accumulation of amyloid-beta-42 (Aβ42) plaques and intracellular accumulation of neurofibrillary tangles (NFTs) that results in microtubule destabilization. Targeted expression of human Aβ42 (GMR > Aβ42) in developing Drosophila eye retinal neurons results in Aβ42 plaque(s) and mimics AD-like extensive neurodegeneration. However, there remains a gap in our understanding of the underlying mechanism(s) for Aβ42-mediated neurodegeneration. To address this gap in information, we conducted a forward genetic screen, and identified N-acetyltransferase 9 (Mnat9) as a genetic modifier of GMR > Aβ42 neurodegenerative phenotype. Mnat9 is known to stabilize microtubules by inhibiting c-Jun-N- …

Cobalt-Sulfur Coordination Chemistry Drives B12 Loading Onto Methionine Synthase, Romila Mascarenhas, Arkajit Guha, Zhu Li, Markus Ruetz, Sojin An, Javier Seravalli, Ruma Banerjee

Department of Biochemistry: Faculty Publications

Cobalt-sulfur (Co-S) coordination is labile to both oxidation and reduction chemistry and is rarely seen in Nature. Cobalamin (or vitamin B12) is an essential cobalt-containing organometallic cofactor in mammals, and is escorted via an intricate network of chaperones to a single cytoplasmic target, methionine synthase. In this study, we report that the human cobalamin trafficking protein, MMADHC, exploits the chemical lability of Co-S coordination, for cofactor off-loading onto methionine synthase. Cys-261 on MMADHC serves as the -axial ligand to cobalamin. Complex formation between MMADHC and methionine synthase is signaled by loss of the lower axial nitrogen ligand, leading to five-coordinate …

A Cost-Effective And Smart Sensing Tissue-Like Testbed For Surgical Training, Lysette Zaragoza

McNair Research Journal SJSU

A low-cost tissue-like testbed with six nodes of varying stiffness was developed for surgical training to provide pressure and force feedback data through image reception to human operators. Using SolidWorks, a 3D model of the box trainer housing was created. A pad for the distribution of smartsensing nodes and microcontroller connections was designed with open spaces for the respective components. The pad was 3D-printed with PLA filament. Flat piezoelectric pressure sensors were fabricated with conductive materials and velostat sensor material. Using static and dynamic analyses, three top sensors were chosen to be used in three pressure sensing nodes. A calibration …

Pyrroline-5-Carboxylate Reductase-2 Promotes Colorectal Carcinogenesis By Modulating Microtubule-Associated Serine/Threonine Kinase-Like/Wnt/Β-Catenin Signaling, Raju Lama Tamang, Balawant Kumar, Sagar M. Patel, Ishwor Thapa, Alshomrani Ahmad, Vikas Kumar, Rizwan Ahmad, Donald F. Becker, Dundy Kiran Bastola, Punita Dhawan, Amar B. Singh

Department of Biochemistry: Faculty Publications

Background: Despite significant progress in clinical management, colorectal cancer (CRC) remains the third most common cause of cancer-related deaths. A positive association between PYCR2 (pyrroline-5-carboxylate reductase-2), a terminal enzyme of proline metabolism, and CRC aggressiveness was recently reported. However, how PYCR2 promotes colon carcinogenesis remains ill understood. Methods: A comprehensive analysis was performed using publicly available cancer databases and CRC patient cohorts. Proteomics and biochemical evaluations were performed along with genetic manipulations and in vivo tumor growth assays to gain a mechanistic understanding. Results: PYCR2 expression was significantly upregulated in CRC and associated with poor patient survival, specifically among PYCR …

Editorial: Rising Stars In Microbial Physiology And Metabolism: 2022, Nicole R. Buan, Ulrike Kappler

Department of Biochemistry: Faculty Publications

This Research Topic was initiated to highlight work by young authors, the rising stars in the field of microbial physiology and metabolism. Microbial physiology and metabolism is an interdisciplinary field of research that seeks to uncover how the metabolic pathways of a cell work together to determine cell fate and function, whether that be growth, replication, pathogenicity, predation, respiration and fermentation, homeostasis or death. Ultimately, researchers like the ones featured here seek to integrate biological information and physicochemical parameters to try to find the underlying rules governing microbial function so that we can understand, predict and design microbes and microbial …

Evaluation Of Bio-Friendly Formulations From Siderophore-Producing Fluorescent Pseudomonas As Biocontrol Agents For The Management Of Soil-Borne Fungi, Fusarium Oxysporum And Rhizoctonia Solani, Gaber Attia Abo-Zaid, Ahmed Salah Abdullah, Nadia Abdel-Mohsen Soliman, Ebaa Ebrahim El-Sharouny, Abdulaziz A. Al-Askar, Yiming Su, Ahmed Abdelkhalek, Soraya Abdel-Fattah Sabry

Civil and Environmental Engineering Faculty Publications

Secretion of siderophores by Pseudomonas aeruginosa F2 and P. fluorescens JY3 was evaluated on chrome azurol S (CAS) agar plates and their inhibitory effect was inspected against Fusarium oxysporum and Rhizoctonia solani. Production of siderophores as biocontrol agents from F2 and JY3 was accomplished in two optimized media. Afterward, cell-free supernatants of the bacterial cultures containing siderophores were used for the preparation of two bio-friendly formulations for the management of F. oxysporum and R. solani under greenhouse conditions. The investigated bacterial isolates, F2 and JY3, showed antagonistic activity in vitro against F. oxysporum and R. solani and produced siderophores …

Chloroquine And Cytosolic Galectins Affect Endosomal Escape Of Antisense Oligonucleotides After Stabilin-Mediated Endocytosis, Ekta Pandey, Ed Harris

Department of Biochemistry: Faculty Publications

Non-DNA-binding Stabilin-2/HARE receptors expressed on liver sinusoidal endothelial cells specifically bind to and internalize several classes of phosphorothioate antisense oligonucleotides (PS-ASOs). After Stabilin-mediated uptake, PS-ASOs are trafficked within endosomes (>97%–99%), ultimately resulting in destruction in the lysosome. The ASO entrapment in endosomes lowers therapeutic efficacy, thereby increasing the overall dose for patients. Here, we use confocal microscopy to characterize the intracellular route transverse by PS-ASOs after Stabilin receptor-mediated uptake in stable recombinant Stabilin-1 and -2 cell lines. We found that PS-ASOs as well as the Stabilin-2 receptor transverse the classic path: clathrincoated vesicle-early endosome-late endosome-lysosome. Chloroquine exposure facilitated endosomal …

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 …