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Distinct Nrf2 Signaling Thresholds Mediate Lung Tumor Initiation And Progression, Janine M. Deblasi

USF Tampa Graduate Theses and Dissertations

NRF2 is a redox-responsive transcription factor the directs the antioxidant program and several critical metabolic processes. Mutations in NRF2 or its negative regulator KEAP1 occur in up to one third of non-small cell lung cancers (NSCLCs) and are often associated with resistance to therapy and poor outcomes. In the present studies, murine alleles of the Keap1 and Nrf2 mutations found in human NSCLC were developed and I comprehensively investigated their impact on tumor initiation and progression. I observed that chronic Nrf2 stabilization by Keap1 loss-of-function or Nrf2 activating mutation was not sufficient to cause lung tumor initiation, even when p53 …

Widefield Imaging Of Rapid Pan-Cortical Voltage Dynamics With An Indicator Evolved For One-Photon Microscopy, Xiaoyu Lu, Yunmiao Wang, Zhuohe Liu, Yueyang Gou, Dieter Jaeger, François St-Pierre

Faculty, Staff and Students Publications

Widefield imaging with genetically encoded voltage indicators (GEVIs) is a promising approach for understanding the role of large cortical networks in the neural coding of behavior. However, the limited performance of current GEVIs restricts their deployment for single-trial imaging of rapid neuronal voltage dynamics. Here, we developed a high-throughput platform to screen for GEVIs that combine fast kinetics with high brightness, sensitivity, and photostability under widefield one-photon illumination. Rounds of directed evolution produced JEDI-1P, a green-emitting fluorescent indicator with enhanced performance across all metrics. Next, we optimized a neonatal intracerebroventricular delivery method to achieve cost-effective and wide-spread JEDI-1P expression in …

Insights Into The Molecular Roles Of Foxr2 In The Pathology Of Primary Pediatric Brain Tumors, Rajendra Gharbaran

Publications and Research

Forkhead box gene R2 (FOXR2) belongs to the family of FOX genes which codes for highly conserved transcription factors (TFs) with critical roles in biological processes ranging from development to organogenesis to metabolic and immune regulation to cellular homeostasis. A number of FOX genes are associated with cancer development and progression and poor prognosis. A growing body of evidence suggests that FOXR2 is an oncogene. Studies suggested important roles for FOXR2 in cancer cell growth, metastasis, and drug resistance. Recent studies showed that FOXR2 is overexpressed by a subset of newly identified entities of embryonal tumors. This review discusses the …

Recent Advances In Crispr/Cas9-Assisted Gene Therapy, Apeksha Srivastava, Shikha Chauhan, Vishal Ahuja

Karbala International Journal of Modern Science

CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats) is an exponentially growing tool with wide-spread applications in therapeutics like gene modifications that focus on altering the hereditary material to repair or eliminate any defective gene-causing diseases like cancer, AIDS (Acquired immunodeficiency syndrome), etc. It also includes the identification of the target sequence with the help of sgRNA followed by the substitution of a malfunction-ing gene with a normal version. It offers high efficiency, specificity, and post-gene-editing efficacy, but have also some off-target impressions, and immunogenic effects. The contribution of CRISPR/Cas9 has already been proved primarily in in-vitro studies using animal germ …

Optical, Structural, And Electrochemical Properties Of P3ht:Y6 Photoactive Layer For Organic Photovoltaics, Hassan Tarikhum B., Furqan Almyahi, Basil Ali

Karbala International Journal of Modern Science

Recently, organic photovoltaics (OPV) have emerged as a promising technology for environmentally friendly energy production. Achieving high performance in OPV requires the discovery of novel compounds. This article aims to study the optical, structural, electrochemical, and electrical properties of a P3HT:Y6 blend. The UV-visible spectra of the blend provided insight into how composition affects the optoelectronic properties of OPV devices. The optimised P3HT:Y6 ratio (0.6:1) resulted in the largest redshift and the highest absorption intensity. Raman and X-ray diffraction tests showed low aggregation and low crystallinity of the polymer P3HT as Y6 content increased. In the OPV devices, the best …

Estimation Of Water Quality Of Dikhu River Of Nagaland Through A Combination Of Water Quality Index And Principal Component Analysis Techniques., Lanuyanger Longchar, Wati Temjen, Khikeya Semy, Wati Lemla

Karbala International Journal of Modern Science

Monitoring the physico-chemical attributes of water is essential for determining water quality. The present study explores the effect of various anthropogenic factors on the quality of Dikhu River, Nagaland. Water samples from three sampling stations were compared against three standards: Indian Council of Medical Research, Bureau of Indian Standards, and World Health Organization. Although all parameters were within the permissible limits, the Water Quality Index categorized all sites during the rainy season as "poor quality", highlighting anthropogenic impacts. A principal component analysis created a Minimum Data Set (MDS) explaining 100%, 93.27%, and 96.26% of the total variance for the three …

Investigating The Influence Of Assessment Question Framing On Undergraduate Biology Student Preference And Affect, Jeremy L. Hsu, Noelle Clark, Kate Hill, Melissa Rowland-Goldsmith

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Nearly all undergraduate biology courses rely on quizzes and exams. Despite their prevalence, very little work has been done to explore how the framing of assessment questions may influence student performance and affect. Here, we conduct a quasi-random experimental study where students in different sections of the same course were given isomorphic questions that varied in their framing of experimental scenarios. One section was provided a description using the self-referential term “you”, placing the student in the experiment; another section received the same scenario that used classmate names; while a third section's scenario integrated counterstereotypical scientist names. Our results demonstrate …

Quantification Of Amu River Riverbank Erosion In Balkh Province Of Afghanistan During 2004–2020, Abdul Basir Mahmoodzada, Divyesh Varade, Sawahiko Shimada, Hiromu Okazawa, Shafiqullah Aryan, Gulbuddin Gulab, Abd El Zaher M.A. Mustafa, Humaira Rizwana, Yogesh Ahlawat, Hosam O. Elansary

Michigan Tech Publications

In this study, we propose quantifying the Amu River riverbank erosion with the modelled river discharge in Kaldar District, Balkh Province of Afghanistan from 2004 to 2020. We propose a framework synergizing multi-source information for modelling the erosion area based on three components: (1) river discharge, (2) river width, and (3) erosion area. The total river discharge for the watershed shared by Afghanistan and Tajikistan was modelled using hydrological parameters from the European Centre for Medium-Range Weather Forecasts (ECMWF) Reanalysis v5 (ERA5) data through multivariate linear regression with ground station data. The river width was determined manually using the Normalized …

Understanding Endangered Sawfish Through The Growth And Structure Of Their Rostral Teeth, Linh Truong, Gabe Gambleton, Jens Hegg

CURCI Student Research

Sawfish are an endangered ray with a toothy rostrum containing a large number of teeth. Largetooth sawfish are a critically endangered species worldwide. Little is known about their life history or behavior, and natural populations are too low to easily track them. Preserved rostra are more accessible to study. Measurements of the rostrum teeth and rostrum length were taken which may indicate a subspecies or sexual dimorphism in the sample. 120 sawfish rostra from the Brazil sample were quantified using ImageJ to compare rostrum size to fish age and length. Sawfish rostral teeth have repeated tubule structures within them that …

Unveiling Novel Avenues In Mtor-Targeted Therapeutics: Advancements In Glioblastoma Treatment, Shilpi Singh, Debashis Barik, Karl Lawrie, Iteeshree Mohapatra, Sujata Prasad, Afsar R. Naqvi, Amar Singh, Gatikrushna Singh

Biology Student Work

The mTOR signaling pathway plays a pivotal and intricate role in the pathogenesis of glioblastoma, driving tumorigenesis and proliferation. Mutations or deletions in the PTEN gene constitutively activate the mTOR pathway by expressing growth factors EGF and PDGF, which activate their respective receptor pathways (e.g., EGFR and PDGFR). The convergence of signaling pathways, such as the PI3K-AKT pathway, intensifies the effect of mTOR activity. The inhibition of mTOR has the potential to disrupt diverse oncogenic processes and improve patient outcomes. However, the complexity of the mTOR signaling, off-target effects, cytotoxicity, suboptimal pharmacokinetics, and drug resistance of the mTOR inhibitors pose …

Avoidance Behavior Ecotoxicity Testing Of Oppia Nitens In Conventional Microplastic And Bioplastic Spiked Soils., Catarina Zabot Pasini, Kaija Carr, Nina Shaffer, Philips Akinwole

Annual Student Research Poster Session

Environment and Climate Change Canada recently completed a standardized protocol of ecotoxicity testing for Oppia nitens, a soil oribatid mite that performs crucial functions to support ecosystem services. Consequently, O. nitens is now recommended among the class of soil invertebrates for ecotoxicity testing, however, its applicability is limited by the paucity of data on its avoidance behavior to contaminants in soils. In this study, we examined the avoidance behavior of O. nitens to three different conventional microplastics (polyvinyl chloride/PVC, polyethylene/PE, polystyrene/PS) and two bioplastics (poly vinyl alcohol/PVA and sodium polyacrylate/NaPa) at five different concentrations following standardized guidelines. PVC and PE …

Stressed, But Doing Fine. An Investigation Of Apocynum Cannabinum Patches And Individuals, Lauren Kyburz, Lily Monnett, Quincie Simmons, Dana Dudle

Annual Student Research Poster Session

Apocynum cannabinum (hemp dogbane) is a native Indiana plant that is abundant in the DePauw University Nature Park’s quarry, a heterogeneous site that contains stretches of dry, rocky terrain in addition to many temporary ponds. In 2023, I mapped the distribution of A. cannabinum in the quarry to see if its habitats are associated with elevation and/or the presence of temporary ponds. I also conducted a study of diurnal pollinator visitation, and an examination of damage caused by the dogbane saucrobotys moth (Saucrobotys futilalis).

Modeling Light For Platanus Occidentalis (American Sycamore) Trees In The Depauw Quarry, Grace Lucchesi, Beth Wilkerson, Quincie Simmons, Dana Dudle

Annual Student Research Poster Session

Platanus Occidentalis, characterized by shedding bark and ball-shaped seed pods, is native to much of the mid-southeastern US, and is commonly used as an ornamental tree. Individuals growing in the quarry of DePauw’s Nature Park are often small, discolored, or even dying. We used devices to measure the UV and Photosynthetically Active Radiation present at the locations of 70 random trees. We then used known sun path equations and GIS to start to develop a way to estimate the maximum amount of time a plant experiences full sun in different locations in the quarry relative to the quarry walls.

Pyrrolidine Derivative Targets Actin Cytoskeleton In Mcf-7 Breast Cancer Cells, An Pham, Jeff Hansen, Sarah Mordan-Mccombs

Annual Student Research Poster Session

Recent research has brought pyrrolidine derivatives into consideration for the development of anticancer drugs with high efficacy and low toxicity. Dr. Hansen’s lab at DePauw has synthesized a pyrrolidine derivative that demonstrated anticancer activity. However, there are many ways a compound can affect cancer cells. In this research, we decided to investigate the mechanism of action of this new compound, specifically on MCF-7 breast cancer cell line. Based on the results, we believe that there is a great likelihood that the pyrrolidine derivative can induce apoptosis (cell death) and disrupt cell movement in MCF-7 cells. In other words, these are …

Life On The Quarry Wall Vs The Quarry Floor: Parthenocissus Quinquefolia, Lily Monnett, Quincie Simmons, Lauren Kyburz

Annual Student Research Poster Session

Parthenocissus quinquefolia, Virginia creeper, is found in two parts of the DePauw Nature Park quarry: vertically along the quarry wall and on large rock piles on the quarry floor. I looked into how these different habitats influenced the growth form, stress responses, and fitness of the P. quinquefolia growing in them. P. quinquefolia on the wall has higher chlorophyll a content and higher water content. There was also a higher percentage of flowering individuals on the quarry wall than on the floor. Lastly, the two habitats had vastly different morphological growth forms, with those on the wall having very long …

Characterizing The Effects Of Benzyl-Amino Alcohol On Cell Growth, Viability, And Migration, Ryan Miller, Jeff Hansen, Sarah Mordan-Mccombs

Annual Student Research Poster Session

The research we are performing investigates a new compound classified as benzyl-amino alcohol and begins a new endeavor into the effects of this class of compound. This compound would work well in chemotherapies because affecting healthy cells can lead to a patient’s health decline.

Effects Of Large Mammal Exclusion On Seedling Communities Depend On Plant Species Traits And Landscape Protection In Human-Modified Costa Rican Forests, Nohemi Huanca-Nuñez, Robin L. Chazdon, Sabrina E. Russo

School of Biological Sciences: Faculty Publications

1. Large terrestrial herbivorous mammals (LTH-mammals) influence plant community structure by affecting seedling establishment in mature tropical forests. Many of these LTH-mammals frequent secondary forests, but their effects on seedling establishment in them are understudied, hindering our understanding of how LTH-mammals influence forest regeneration in human-modified landscapes.

2. We tested the hypothesis that the strength of LTH-mammals' effects on seedling establishment depends on landscape protection, forest successional stage and plant species' traits using a manipulative field experiment in six 1-ha sites with varying successional age and landscape protection. In each site, we established 40 seedling plot-pairs, with one plot excluding LTH-mammals and …

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Effects Of Large Mammal Exclusion On Seedling Communities Depend On Plant Species Traits And Landscape Protection In Human-Modified Costa Rican Forests, Nohemi Huanca-Nuñez, University Of Connecticut, University Of The Sunshine Coast, Sabrina E. Russo

School of Biological Sciences: Faculty Publications

1. Large terrestrial herbivorous mammals (LTH-mammals) influence plant community structure by affecting seedling establishment in mature tropical forests. Many of these LTH-mammals frequent secondary forests, but their effects on seedling establishment in them are understudied, hindering our understanding of how LTH-mammals influence forest regeneration in human-modified landscapes.
2. We tested the hypothesis that the strength of LTH-mammals' effects on seedling establishment depends on landscape protection, forest successional stage and plant species' traits using a manipulative field experiment in six 1-ha sites with varying successional age and landscape protection. In each site, we established 40 seedling plot-pairs, with one plot excluding LTH-mammals and …

Gaxin2-Xo3 Surface Pyramids Interaction With Formaldehyde: Thermodynamic And Sensing Analysis, Mudar Ahmed Abdulsattar

Karbala International Journal of Modern Science

GaxIn2-xO3 surface pyramids' electronic structures are investigated using density functional theory, including dispersion corrections. Application of GaxIn2-xO3 surface pyramids as a gas sensor for formaldehyde is also performed and compared with experimental findings. These findings show that the energy gap of these pyramids follows closely with the bulk values. The energy gap increases between the two limits, In2O3 and Ga2O3. Applying GaxIn2-xO3 surface pyramids as a gas sensor uses transition state theory formalism. Thermodynamic quantities such as activation Gibbs energy, enthalpy, and entropy are needed for temperature-dependent calculations. A comparison of sensor response which is proportional to reaction rate as …

Seed Coatings Containing Capsaicin Reduce Seed Removal In Temperate Woody Species, Brian M. Connolly, Francesca Lanni, John L. Orrock, Peter W. Guiden

Biology Faculty Scholarship

Coating seeds in taste-averting chemical defenses (e.g., capsaicin from Capsicum spp.) can reduce rodent seed predation, and thus might promote plant establishment and forest regeneration. However, the efficacy of such seed coatings remains unknown for many woody plant species, and seed coatings have not been evaluated across different habitats where forest managers might seek to promote forest regeneration. We used two complementary seed-removal experiments in closed-canopy forests (Michigan) and an old field undergoing reforestation (New York) to examine whether coating seeds of four native tree species (Acer rubrum, Fagus grandifolia, Pinus banksiana, and Pinus resinosa) with chili …

Reimagining Large River Management Using The Resist–Accept–Direct (Rad) Framework In The Upper Mississippi River, Nicole K. Ward, Abigail J. Lynch, Erik A. Beever, Joshua Booker, Kristen L. Bouska, Holly Embke, Jeffrey N. Houser, John F. Kocik, David J. Lawrence, Mary Grace Lemon, Doug Limpinsel, Madeline R. Magee, Bryan M. Maitland, Owen Mckenna, Andrew Meier, John M. Morton, Jeffrey D. Muehlbauer, Robert Newman, Devon C. Oliver, Heidi M. Rantala, Greg G. Sass, Aaron Shultz, Laura M. Thompson, Jennifer L. Wilkening

Biology Faculty Publications

Background Large-river decision-makers are charged with maintaining diverse ecosystem services through unprecedented social-ecological transformations as climate change and other global stressors intensify. The interconnected, dendritic habitats of rivers, which often demarcate jurisdictional boundaries, generate complex management challenges. Here, we explore how the Resist–Accept–Direct (RAD) framework may enhance large-river management by promoting coordinated and deliberate responses to social-ecological trajectories of change. The RAD framework identifies the full decision space of potential management approaches, wherein managers may resist change to maintain historical conditions, accept change toward different conditions, or direct change to a specified future with novel conditions. In the Upper Mississippi …

Spa-Stocsy: An Automated Tool For Identifying Annotated And Non-Annotated Metabolites In High-Throughput Nmr Spectra, Xu Han, Wanli Wang, Li-Hua Ma, Ismael Ai-Ramahi, Juan Botas, Kevin Mackenzie, Genevera I Allen, Damian W Young, Zhandong Liu, Mirjana Maletic-Savatic

Faculty, Staff and Students Publications

MOTIVATION: Nuclear magnetic resonance spectroscopy (NMR) is widely used to analyze metabolites in biological samples, but the analysis requires specific expertise, it is time-consuming, and can be inaccurate. Here, we present a powerful automate tool, SPatial clustering Algorithm-Statistical TOtal Correlation SpectroscopY (SPA-STOCSY), which overcomes challenges faced when analyzing NMR data and identifies metabolites in a sample with high accuracy.

RESULTS: As a data-driven method, SPA-STOCSY estimates all parameters from the input dataset. It first investigates the covariance pattern among datapoints and then calculates the optimal threshold with which to cluster datapoints belonging to the same structural unit, i.e. the metabolite. …

Scribbledom: Using Scribble-Annotated Histology Images To Identify Domains In Spatial Transcriptomics Data, Mohammad Nuwaisir Rahman, Abdullah Al Noman, Abir Mohammad Turza, Mohammed Abid Abrar, Md Abul Hassan Samee, M Saifur Rahman

Faculty, Staff and Students Publications

Motivation

Spatial domain identification is a very important problem in the field of spatial transcriptomics. The state-of-the-art solutions to this problem focus on unsupervised methods, as there is lack of data for a supervised learning formulation. The results obtained from these methods highlight significant opportunities for improvement.

Results

In this article, we propose a potential avenue for enhancement through the development of a semi-supervised convolutional neural network based approach. Named “ScribbleDom”, our method leverages human expert’s input as a form of semi-supervision, thereby seamlessly combines the cognitive abilities of human experts with the computational power of machines. ScribbleDom incorporates a …

Kinome Reprogramming Is A Targetable Vulnerability In Esr1 Fusion-Driven Breast Cancer, Xuxu Gou, Beom-Jun Kim, Meenakshi Anurag, Jonathan T Lei, Meggie N Young, Matthew V Holt, Diana Fandino, Craig T Vollert, Purba Singh, Mohammad A Alzubi, Anna Malovannaya, Lacey E Dobrolecki, Michael T Lewis, Shunqiang Li, Charles E Foulds, Matthew J Ellis

Faculty, Staff and Students Publications

Transcriptionally active ESR1 fusions (ESR1-TAF) are a potent cause of breast cancer endocrine therapy (ET) resistance. ESR1-TAFs are not directly druggable because the C-terminal estrogen/anti-estrogen-binding domain is replaced with translocated in-frame partner gene sequences that confer constitutive transactivation. To discover alternative treatments, a mass spectrometry (MS)-based kinase inhibitor pulldown assay (KIPA) was deployed to identify druggable kinases that are upregulated by diverse ESR1-TAFs. Subsequent explorations of drug sensitivity validated RET kinase as a common therapeutic vulnerability despite remarkable ESR1-TAF C-terminal sequence and structural diversity. Organoids and xenografts from a pan-ET-resistant patient-derived xenograft model that harbors the ESR1-e6>YAP1 TAF were …

Chronic Kidney Disease Promotes Atrial Fibrillation Via Inflammasome Pathway Activation, Jia Song, Jose Alberto Navarro-Garcia, Jiao Wu, Arnela Saljic, Issam Abu-Taha, Luge Li, Satadru K Lahiri, Joshua A Keefe, Yuriana Aguilar-Sanchez, Oliver M Moore, Yue Yuan, Xiaolei Wang, Markus Kamler, William E Mitch, Gema Ruiz-Hurtado, Zhaoyong Hu, Sandhya S Thomas, Dobromir Dobrev, Xander Ht Wehrens, Na Li

Faculty, Staff and Students Publications

Chronic kidney disease (CKD) is associated with a higher risk of atrial fibrillation (AF). The mechanistic link between CKD and AF remains elusive. IL-1β, a main effector of NLR family pyrin domain-containing 3 (NLRP3) inflammasome activation, is a key modulator of conditions associated with inflammation, such as AF and CKD. Circulating IL-1β levels were elevated in patients with CKD who had AF (versus patients with CKD in sinus rhythm). Moreover, NLRP3 activity was enhanced in atria of patients with CKD. To elucidate the role of NLRP3/IL-1β signaling in the pathogenesis of CKD-induced AF, Nlrp3-/- and WT mice were subjected to …

Heterocyclic Merging Of Stereochemically Diverse Chiral Piperazines And Morpholines With Indazoles, Amol B Viveki, Timothy M Mansfield, Kevin A Tran, Evan Lenkeit, Kevin R Mackenzie, Damian W Young, Srinivas Chamakuri

Faculty, Staff and Students Publications

We report a heterocyclic merging approach to construct novel indazolo-piperazines and indazolo-morpholines. Starting from chiral diamines and amino alcohols, novel regiochemically (1,3 and 1,4) and stereochemically diverse (relative and absolute) cohorts of indazolo-piperazines and indazolo-morpholines were obtained within six or seven steps. The key transformations involved are a Smiles rearrangement to generate the indazole core structure and a late-stage Michael addition to build the piperazine and morpholine heterocycles. We further explored additional vector diversity by incorporating substitutions on the indazole aromatic ring, generating a total of 20 unique, enantiomerically pure heterocyclic scaffolds.

Meiosis In Budding Yeast, G. Valentin Borner, Andreas Hochwagen, Amy J. Macqueen

Biological, Geological, and Environmental Faculty Publications

Meiosis is a specialized cell division program that is essential for sexual reproduction. The two meiotic divisions reduce chromosome number by half, typically generating haploid genomes that are packaged into gametes. To achieve this ploidy reduction, meiosis relies on highly unusual chromosomal processes including the pairing of homologous chromosomes, assembly of the synaptonemal complex, programmed formation of DNA breaks followed by their processing into crossovers, and the segregation of homologous chromosomes during the first meiotic division. These processes are embedded in a carefully orchestrated cell differentiation program with multiple interdependencies between DNA metabolism, chromosome morphogenesis, and waves of gene expression …

Warmer Temperature Overrides The Effects Of Antidepressants On Amphibian Metamorphosis And Behavior, Peter Fong, Aylin Doganoglu, Eleanor Sandt, Sierra Turbeville

Biology Faculty Publications

Climate change can exacerbate the effects of environmental pollutants on aquatic organisms. Pollutants such as human antidepressants released from wastewater treatment plants have been shown to impact life-history traits of amphibians. We exposed tadpoles of the wood frog Lithobates sylvaticus to two temperatures (20°C and 25°C) and two antidepressants (fluoxetine and venlafaxine), and measured timing of metamorphosis, mass at metamorphosis, and two behaviors (startle response and percent motionless). Antidepressants significantly shortened time to metamorphosis at 20°C, but not at 25°C. At 25°C, tadpoles metamorphosed significantly faster than those at 20°C independent of antidepressant exposure. Venlafax- ine reduced body mass at …

Abalone Resource Of Western Australia: Harvest Strategy: 2021-2026 : Version 2.2, Department Of Primary Industries And Regional Development, Western Australia

Fisheries Management Papers

This harvest strategy has been developed and revised in line with the Department’s Harvest Strategy Policy for Aquatic Resources (Department of Fisheries 2015) and is consistent with relevant national harvest strategy policies and guidelines (e.g. Sloan et al. 2014; Department of Agriculture and Water Resources 2018a, b). It makes explicit the performance indicators, reference levels, and harvest control rules designed to achieve the specific long- and short-term management objectives for the resource, and the broader goals of ESD and EBFM.

The publication of this harvest strategy is intended to make the decision-making considerations and processes for the management of specified …

Determining The Structure And Function Of A Bacteriophage Called Leozinho, Leonardo Rodrigues Venturotti

Biology Student Work

A research team, having discovered a bacteriophage named Leozinho a couple of years ago, embarked on a mission to unravel the structure of an unidentified gene, gp_61. They utilized ESMatlas, Foldseek, and ChimeraX to predict the structure of gp_61 and afterwards various laboratory procedures were employed to facilitate the growth of the gene. Ultimately, the experiment was concluded with the successful cloning of gp_61 into a plasmid vector for protein expression.