Digital Commons Network^™

Generation Of A Wt1 Conditional Deletion, Nuclear Red Fluorescent Protein Reporter Allele In The Mouse, Jace A Aloway, E Cristy Ruteshouser, Vicki Huff, Richard R Behringer

Faculty, Staff and Student Publications

A Wt1 conditional deletion, nuclear red fluorescent protein (RFP) reporter allele was generated in the mouse by gene targeting in embryonic stem cells. Upon Cre-mediated recombination, a deletion allele is generated that expresses RFP in a Wt1-specific pattern. RFP expression was detected in embryonic and adult tissues known to express Wt1, including the kidney, mesonephros, and testis. In addition, RFP expression and WT1 co-localization was detected in the adult uterine stroma and myometrium, suggesting a role in uterine function. Crosses with Wnt7a-Cre transgenic mice that express Cre in the Müllerian duct epithelium activate Wt1-directed RFP expression in the epithelium of …

Pregnancy-Related Factors Responsible For Delivering Low Birth Weight Babies: An Institutional-Based Cross-Sectional Study, Jamnagar, Gujarat, Harsh K Patel, Jitesh P Mehta, Bela A Patel, Rohitkumar Ram, Dipesh V Parmar

Faculty, Staff and Student Publications

BACKGROUND: The WHO defines LBW as "Birth weight less than 2500 grams" regardless of gestational age. Being born with a low birth weight also incurs enormous economic costs, including higher medical expenditures and social service expenses, and decreased productivity in adulthood.

OBJECTIVE: To study distribution of newborns' according to pregnancy related factors and its association with newborns' birth weight.

METHODS: An institutional based cross-sectional study. New-borns delivered at study institute were considered as study participants. Estimated final sample size was 500. Guardians (mothers) were face-to-face interviewed and also recorded data were collected from the case file and Mother and Child …

A Goldilocks Computational Protocol For Inhibitor Discovery Targeting Dna Damage Responses Including Replication-Repair Functions, Davide Moiani, John A Tainer

Faculty, Staff and Student Publications

While many researchers can design knockdown and knockout methodologies to remove a gene product, this is mainly untrue for new chemical inhibitor designs that empower multifunctional DNA Damage Response (DDR) networks. Here, we present a robust Goldilocks (GL) computational discovery protocol to efficiently innovate inhibitor tools and preclinical drug candidates for cellular and structural biologists without requiring extensive virtual screen (VS) and chemical synthesis expertise. By computationally targeting DDR replication and repair proteins, we exemplify the identification of DDR target sites and compounds to probe cancer biology. Our GL pipeline integrates experimental and predicted structures to efficiently discover leads, allowing …

Aging-Related Biomarker Discovery In The Era Of Immune Checkpoint Inhibitors For Cancer Patients, Abdullah Al-Danakh, Mohammed Safi, Yuli Jian, Linlin Yang, Xinqing Zhu, Qiwei Chen, Kangkang Yang, Shujing Wang, Jianjun Zhang, Deyong Yang

Faculty, Staff and Student Publications

Older patients with cancer, particularly those over 75 years of age, often experience poorer clinical outcomes compared to younger patients. This can be attributed to age-related comorbidities, weakened immune function, and reduced tolerance to treatment-related adverse effects. In the immune checkpoint inhibitors (ICI) era, age has emerged as an influential factor impacting the discovery of predictive biomarkers for ICI treatment. These age-linked changes in the immune system can influence the composition and functionality of tumor-infiltrating immune cells (TIICs) that play a crucial role in the cancer response. Older patients may have lower levels of TIICs infiltration due to age-related immune …

Distinct Reproductive Phenotypes Segregate With Differences In Body Weight In Adolescent Polycystic Ovary Syndrome., Angie Chen-Patterson, Angelina Bernier, Tania S. Burgert, Vanessa Davis, Tazeena Khan, David Geller, Emily Paprocki, Rachana Shah, Selma F. Witchel, Camila Pereira-Eshraghi, Aviva B. Sopher, Melanie G. Cree, Laura C. Torchen

Manuscripts, Articles, Book Chapters and Other Papers

INTRODUCTION: Polycystic ovary syndrome (PCOS) is a heterogenous clinical syndrome defined by hyperandrogenism and irregular menses. In adult women with PCOS, discrete metabolic and reproductive subgroups have been identified. We hypothesize that distinct phenotypes can be distinguished between adolescent girls who are lean (LN-G) and girls with obesity (OB-G) at the time of PCOS diagnosis.

METHODS: Data were extracted from the CALICO multisite PCOS database. Clinical data collected at the time of diagnosis were available in 354 patients (81% with obesity) from 7 academic centers. Patients with body mass index (BMI) < 85th percentile for age and sex were characterized as lean (LN-G) and those with BMI percentile ≥ 95th percentile as obese (OB-G). We compared metabolic and reproductive phenotypes in LN-G and OB-G.

RESULTS: Reproductive phenotypes differed between the groups, with LN-G …

Activation Of Piezo1 Increases The Sensitivity Of Breast Cancer To Hyperthermia Therapy, Shao-Kang Wang, Xiao-Ting Zhang, Xuan-Yao Jiang, Bi-Jiang Geng, Tao-Lin Qing, Lei Li, Yun Chen, Jin-Feng Li, Xiao-Fang Zhang, Shuo-Gui Xu, Jiang-Bo Zhu, Yu-Ping Zhu, Mei-Tang Wang, Ji-Kuai Chen

Faculty, Staff and Student Publications

Photothermal therapy (PTT) of nanomaterials is an emerging novel therapeutic strategy for breast cancer. However, there exists an urgent need for appropriate strategies to enhance the antitumor efficacy of PTT and minimize damage to surrounding normal tissues. Piezo1 might be a promising novel photothermal therapeutic target for breast cancer. This study aims to explore the potential role of Piezo1 activation in the hyperthermia therapy of breast cancer cells and investigate the underlying mechanisms. Results showed that the specific agonist of Piezo1 ion channel (Yoda1) aggravated the cell death of breast cancer cells triggered by heat stress

Chronic Adolescent Stress Alters Gr-Fkbp5 Interactions In The Hippocampus Of Adult Female Rats, Sydney Rowson, Mandakh Bekhbat, Sean Kelly, Molly M Hyer, Samya Dyer, David Weinshenker, Gretchen Neigh

Faculty, Staff and Student Publications

Chronic stress exposure during development can have lasting behavioral consequences that differ in males and females. More specifically, increased depressive behaviors in females, but not males, are observed in both humans and rodent models of chronic stress. Despite these known stress-induced outcomes, the molecular consequences of chronic adolescent stress in the adult brain are less clear. The stress hormone corticosterone activates the glucocorticoid receptor, and activity of the receptor is regulated through interactions with co-chaperones-such as the immunophilin FK506 binding proteins 5 (FKBP5). Previously, it has been reported that the adult stress response is modified by a history of chronic …

Editorial: Genetic And Cellular Heterogeneity In Tumors, Zishan Wang, Li Ma, Juan Xu, Chunjie Jiang

Faculty, Staff and Student Publications

No abstract provided.

Egfr, Hla-G, Cd70, C-Met, And Ny-Eso1 As Potential Biomarkers In High Grade Epithelial Ovarian Carcinoma, Duc Vo, Yan Liu, Anil K Sood, Katy Rezvani, Amir A Jazaeri, Jinsong Liu

Faculty, Staff and Student Publications

High grade epithelial ovarian carcinoma is an aggressive tumor. Treatment includes platinum therapy, however it recurs in most patients due to therapy resistance. In this project, we study the immunohistochemical (IHC) expression of five potential biomarkers/prognostic markers in high grade epithelial ovarian carcinoma: EGFR, HLA-G, CD70, c-MET, and NY-ESO1. A cohort of 274 patients is used. We compare the IHC expression with age, stage, ascites status, family history of cancer, disease free survival (DFS) and overall survival (OS). EGFR expression is significantly correlated with family history and worse OS. HLA-G is associated with worse OS. To confirm the results of …

Willingness To Accept Monkeypox Vaccine And Its Correlates Among Men Who Have Sex With Men In Southern China: A Web-Based Online Cross-Sectional Study, Xinju Huang, Zhifeng Lin, Jiao Qin, Dee Yu, Fei Zhang, Ganggang Fang, Xi Chen, Jinfeng He, Ping Cen, Mu Li, Rongjing Zhang, Tong Luo, Junjun Jiang, Sanqi An, Hao Liang, Li Ye, Bingyu Liang

Faculty, Staff and Student Publications

BACKGROUND: The May 2022 global outbreak of monkeypox (MPX) poses a threat to the health of men who have sex with men. However, there is limited data on the willingness of MSM to receive monkeypox vaccination in Southern China. This study aimed to assess the knowledge of MPX, concerns regarding MPX, and willingness to receive monkeypox vaccination, as well as their correlates, among MSM in China.

METHODS: We conducted a Web-based online survey of MSM in Southern China from August to September 2022. Data were collected on the socio-demographic characteristics, knowledge, worries, concerns regarding MPX and willingness to receive monkeypox …

Lactulose Vs Polyethylene Glycol For Bowel Preparation: A Single-Center, Prospective, Randomized Controlled Study Based On Bmi, Sun Wenqi, Zhao Bei, Wang Yunrong, Wu Xinrong, Li Yifan, Wang Weiwei, Ni Muhan, Yan Peng, Dou Xiaotan, Li Wen, Yang Hua, Wang Lei, Chen Min

Faculty, Staff and Student Publications

INTRODUCTION: Colonoscopy is currently considered as one of the principal techniques to diagnose the colorectal diseases. Admittedly, qualified bowel preparation before colonoscopy is a premise for high-quality examination. Lower quality bowel preparation might seriously impede visualization of the intestinal mucosa, resulting in missed and misdiagnosed intestinal lesions. Therefore, it is necessary to choose the appropriate oral laxative based on the guarantee of safety and efficacy.

METHODS: This prospective randomized controlled study was conducted to compare lactulose oral solution and polyethylene glycol (PEG) electrolyte powder for bowel preparation using the following indicators: Boston Bowel Preparation Scale, Bowel Bubble Score, detection rate …

Covid-19 Convalescent Plasma Therapy: Long-Term Implications, Hyunah Yoon, Yi Li, Keith S Goldfeld, Gia F Cobb, Caroline L Sturm-Reganato, Luis Ostrosky-Zeichner, Dushyantha T Jayaweera, Julie V Philley, Mahalia S Desruisseaux, Marla J Keller, Judith S Hochman, Liise-Anne Pirofski, Mila B Ortigoza

Faculty, Staff and Student Publications

BACKGROUND: The long-term effect of coronavirus disease 2019 (COVID-19) acute treatments on postacute sequelae of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection (PASC) is unknown. The CONTAIN-Extend study explores the long-term impact of COVID-19 convalescent plasma (CCP) therapy on postacute sequelae of SARS-CoV-2 infection (PASC) symptoms and general health 18 months following hospitalization.

METHODS: The CONTAIN-Extend study examined 281 participants from the original CONTAIN COVID-19 trial (CONTAIN-RCT, NCT04364737) at 18 months post-hospitalization for acute COVID-19. Symptom surveys, global health assessments, and biospecimen collection were performed from November 2021 to October 2022. Multivariable logistic and linear regression estimated associations between …

Predicting Suicidality In Late-Life Depression By 3d Convolutional Neural Network And Cross-Sample Entropy Analysis Of Resting-State Fmri, Chemin Lin, Chih-Mao Huang, Wei Chang, You-Xun Chang, Ho-Ling Liu, Shu-Hang Ng, Huang-Li Lin, Tatia Mei-Chun Lee, Shwu-Hua Lee, Shun-Chi Wu

Faculty, Staff and Student Publications

BACKGROUND: Predicting suicide is a pressing issue among older adults; however, predicting its risk is difficult. Capitalizing on the recent development of machine learning, considerable progress has been made in predicting complex behavior such as suicide. As depression remained the strongest risk for suicide, we aimed to apply deep learning algorithms to identify suicidality in a group with late-life depression (LLD).

METHODS: We enrolled 83 patients with LLD, 35 of which were non-suicidal and 48 were suicidal, including 26 with only suicidal ideation and 22 with past suicide attempts, for resting-state functional magnetic resonance imaging (MRI). Cross-sample entropy (CSE) analysis …

Probing Membrane-Associated Cytoskeletal Oligomers Of The Bacterial Divisome By Electron Microscopy And Tomography, Bo Hu, William Margolin

Faculty, Staff and Student Publications

The cell division machinery or "divisome" of many bacteria, including Escherichia coli, contains homologs of tubulin (FtsZ) and actin (FtsA) that interact with each other to promote the synthesis of septal peptidoglycan. FtsA oligomers have an essential role as a track for tethering dynamically treadmilling FtsZ protofilaments to the cytoplasmic membrane. Other bacterial cytoskeletal oligomers such as MreB also assemble on and move along the membrane. Structures of these oligomers on membranes in vitro may mimic their behavior in the cell. Here, we describe a protocol to visualize FtsA oligomeric structures on membranes and their interactions with FtsZ protofilaments using …

In Sickness And In Health-Type I Interferon And The Brain, Wei Cao

Faculty, Staff and Student Publications

Type I interferons (IFN-I) represent a group of pleiotropic cytokines renowned for their antiviral activity and immune regulatory functions. A multitude of studies have unveiled a critical role of IFN-I in the brain, influencing various neurological processes and diseases. In this mini-review, I highlight recent findings on IFN-I's effects on brain aging, Alzheimer's disease (AD) progression, and central nervous system (CNS) homeostasis. The multifaceted influence of IFN-I on brain health and disease sheds light on the complex interplay between immune responses and neurological processes. Of particular interest is the cGAS-STING-IFN-I axis, which extensively participates in brain aging and various forms …

Shipwrecks Act As De Facto Marine Protected Areas In Areas Of Heavy Fishing Pressure, Jenny Hickman, Joe Richards, Adam Rees, Emma V. Sheehan

School of Biological and Marine Sciences

Ubiquitous, industrial use of bottom‐towed fishing gear since the 1800s has altered marine communities and ecosystem services. Outside of legal protection, only areas inaccessible to trawlers are offered any protection. Shipwrecks present hazards to fishing gear and are rarely subject to trawling pressure. As many have been in situ for >100 years, they offer a baseline of ecological potential when trawling pressure is reduced or removed. Five shipwrecks were surveyed off the Berwickshire coast, some within the Static Gear Reserve and others outside. Video transects were collected of shipwrecks, the surrounding 50 m radius and control locations >150 m from …

The Impacts Of Crassula Helmsii On Northwest European Freshwater Macroinvertebrates, Samuel Tasker

School of Biological and Marine Sciences Theses

Crassula helmsii (New Zealand pygmyweed) is an Australasian aquatic plant which has invaded European freshwaters since the 1950s, provoking widespread concern amongst conservationists. The ecological impacts of C. helmsii invasion remain unclear, however, particularly with respect to macroinvertebrates. The main aim of this thesis is therefore to determine how Crassula helmsii impacts recipient macroinvertebrate assemblages in small lentic waterbodies across its invasive range. To this end, I surveyed small lentic waterbodies across the UK, Belgium and the Netherlands, including invaded and ecologically similar uninvaded sites in each region, assessing the impacts of C. helmsii invasion on macroinvertebrates using a suite …

Nuanced Impacts And Social Perceptions Of Alpine Newts Ichthyosaura Alpestris In The Uk, Alexandra North

School of Biological and Marine Sciences Theses

Invasive species are major drivers of environmental change and their incidence appear to be growing worldwide. An understanding of the subtleties and complexities of invasion is important for conservation decision-making as not all introduced species become invasive, and the impacts of those that do can be highly context dependent. The alpine newt Ichthyosaura alpestris is a non-native species that is now widely distributed across the UK. An understanding of the drivers of introduction and spread of this species are in its infancy, and whilst anecdotal reports are indicative of negative ecological impacts, empirical evidence is lacking. This interdisciplinary thesis aims …

Environmental Filtering In The Anthropocene: Investigating Community Assembly Processes At Multiple Scales, Charlotte Clubley

School of Biological and Marine Sciences Theses

Environmental change in the Anthropocene is driving the redistribution of species and restructuring of communities globally. In marine systems, anthropogenic impacts, including the deliberate and accidental movement of species (i.e., non-native and invasive species) and construction of artificial structures (i.e., ocean sprawl), are creating novel opportunities for range expansion, altering the abiotic environment and causing global biodiversity loss. As the geographic distribution and community assembly of species depends on dispersal ability, abiotic tolerance and interactions with other species, understanding the effects that anthropogenic impacts will have on these processes is critical to predicting how ecosystem structure and function is likely …

Computer Vision Enabled Approaches To Predicting Responses To Fluctuating Temperatures In Aquatic Embryos, Ziad Ibbini

School of Biological and Marine Sciences Theses

Fluctuations in temperature are prevalent within environments inhabited by many ectotherms, and climate change continues to rapidly alter the thermal regimes of these environments. Early developmental stages are often the most thermally sensitive life stage, and thus can drive a species’ vulnerability to climate change. Our current understanding of the effects of fluctuating temperatures (FTs) is predominately based upon comparisons of responses to constant temperatures (CTs). Relating CT responses to those of FTs is typically achieved through the use of thermal performance curves, which are integrated over time to estimate cumulative performance and thus a developmental endpoint; an approach often …

Cas Monthly Newsletter January 2024, Andrews University

College of Arts & Sciences Newsletter

Key Dates for January

IN THIS ISSUE

UPDATES & REPORTS

Welcome to Dr. Jill Manoukian

Religion Department News

Visual Art, Communication & Design Department News

Community & International Development Program News

HAPPENINGS

Quantitative Research Working Group

Faculty & Staff Appreciation Week

Winter Symposium

Christmas at the Biology Botanical Conservatory

ACADEMIC & PROFESSIONAL DEVELOPMENT

Research Deadlines

ADDITIONAL RESOURCES

Maker Lab

Writing Center

Registration Tips

New Copy Machine in Nethery Hall

The Food And Habitat Effects Of Detrital Kelp As A Trophic Subsidy To Seagrass-Inhabiting Mesograzers, Jessica Billinghurst

Theses: Doctorates and Masters

Seascapes are connected through the transfer of energy from the movement of fauna, detritus, and nutrients among habitats. This exchange of resources can influence food web dynamics in the recipient system, increasing productivity and biodiversity, thus creating spatial subsidies. Spatial subsidies are usually characterised by the transfer of resources from highly productive donor systems to lower productive recipient systems. However, recent work suggests that recipient systems with comparably high productivity, such as seagrass meadows, may benefit from spatial subsidies. Kelp is regularly eroded from reefs and provides a vector for nutrient and detrital transfer to seagrass meadows, where it can …

Perceptions Towards Integrating A Gender Perspective Into Education: A Study Of Muslim Preservice Teachers In The Philippines, Veronico N. Tarrayo, Ma. Annie Susaya, Delma A. Yuarata

Australian Journal of Teacher Education

: To date, studies exploring Muslim preservice teachers’ views on gender matters in education remain scant. We conducted a qualitative-quantitative survey with 45 Filipino Muslim preservice teachers, ten of whom participated in follow-up email interviews, to explore their perceptions towards integrating a gender perspective into education. The results indicate the participants’ progressive views on fluid gender norms, although they also share conflicting perspectives on gender roles that tend to reinforce gender stereotypes. The preservice teachers also report positive perceptions concerning their institutions’ commitment to gender sensitivity and gender mainstreaming in teacher-education curricula. However, they seem to have unrealistic views on …

Review On Sizing And Management Of Stand-Alone Pv/Wind Systems With Storage, Guétinsom Jean Kafando, Daniel Yamegueu, Etienne Tchoffo Houdji

All Rsif Scholars' Publications

Extending the public electricity grid to rural or peri-urban areas is sometimes very costly and unprofitable due to their remoteness, low population density and sometimes difficult accessibility. In view of this, and in the concern of a sustainable development, the autonomous PV and/or wind power systems is increasingly used. However, these fluctuating source systems remain unreliable due especially to their intermittent nature, what justifies the integration of battery storage systems to them. They are also still expensive, particularly in the African context, limiting their access to the greatest number of the population. In addition to these problems of cost and …

Genomic Epidemiology Of Rift Valley Fever Virus Involved In The 2018 And 2022 Outbreaks In Livestock In Rwanda, Isidore Nsengimana, John Juma, Kristina Roesel, Methode N. Gasana, Fabrice Ndayisenga, Claude M. Muvunyi, Emmanuel Hakizimana, Jean N. Hakizimana, Gillian Eastwood, Augustino A. Chengula, Bernard Bett, Christopher J. Kasanga, Samuel O. Oyola

All Rsif Scholars' Publications

Rift Valley fever (RVF), a mosquito-borne transboundary zoonosis, was first confirmed in Rwanda’s livestock in 2012 and since then sporadic cases have been reported almost every year. In 2018, the country experienced its first large outbreak, which was followed by a second one in 2022. To determine the circulating virus lineages and their ancestral origin, two genome sequences from the 2018 outbreak, and thirty-six, forty-one, and thirty-eight sequences of small (S), medium (M), and large (L) genome segments, respectively, from the 2022 outbreak were generated. All of the samples from the 2022 outbreak were collected from slaughterhouses. Both maximum likelihood …

Challenging The Criminalization Of Undocumented Drivers Through A Health-Justice Framework, Jason A. Cade

Scholarly Works

States increasingly use driver’s license laws to further policy objectives unrelated to road safety. This symposium contribution employs a health justice lens to focus on one manifestation of this trend—state schemes that prohibit noncitizen residents from accessing driver’s licenses and then impose criminal sanctions for driving without authorization. Status-based no-license laws not only facilitate legally questionable enforcement of local immigration priorities but also impose structural inequities with long-term health consequences for immigrants and their family members, including US citizen children. Safe, reliable transportation is a significant social determinant of health for individuals, families, and communities. Applying a health justice lens …

Music And Perceived Stress: An Investigation Into The Effects Of Music On Chemistry Students' Perceived Stress Levels, Alice Young, Eric Malina

Honors Program: Senior Projects (Public)

Music has long been a prevalent intervention when trying to lower stress in certain populations (Thoma et al., 2013). This study aimed to explore the possible usefulness of music as an intervention for students experiencing stress in the chemistry laboratory setting. Students in general chemistry laboratories were surveyed regarding their stress at the ends of periods in which music was or was not played in their laboratory classes. While the results were not statistically significant, mean stress scores did lower in those groups where music was played. Further research into this topic should focus on type of music, the effects …

The Consequences Of Mythology: Supreme Court Decisionmaking In Indian Country, M. Alexander Pearl

Faculty Articles

Ilanoli isht unowa. We tell our own stories. A single historical event has many stories. Although this nation's official chronicle expected and even hoped for Indigenous peoples to fade away, we are still here. Our histories are marked by resistance, survival, sovereignty, and renaissance. Only now, in the later stages of the American experiment, do our histories have the chance to matter in new forms and spaces. How much these stories matter within contemporary contexts depends upon where they are spoken and more importantly, who is listening. On the pages of a U.S. Supreme Court opinion, what stories are told …

Enhancing Identification Of Meteorological And Biological Targets Using The Depolarization Ratio For Weather Radar: A Case Study Of Faw Outbreak In Rwanda, Fidele Maniraguha, Anthony Vodacek, Kwang Soo Kim, Emmanuel Ndashimye, Gerard Rushingabigwi

All Rsif Scholars' Publications

Leveraging weather radar technology for environmental monitoring, particularly the detection of biometeors like birds, bats, and insects, presents a significant challenge due to the dynamic nature of their behavior. Unlike hydrometeor targets, biometeor targets exhibit arbitrary changes in direction and position, which significantly alter radar wave polarization upon scattering. This study addresses this challenge by introducing a novel methodology utilizing Rwanda’s C-Band Polarization Radar. Our approach exploits the capabilities of dual-polarization radar by analyzing parameters such as differential reflectivity (ZDR) and correlation coefficient (RHOHV) to derive the Depolarization Ratio (DR). While existing radar metrics offer valuable insights, they have limitations …

Enhanced Neonatal Screening For Sickle Cell Disease: Human-Guided Deep Learning With Cnn On Isoelectric Focusing Images, Kpangni Alex Jérémie Koua, Cheikh Talibouya Diop, Lamine Diop, Mamadou Diop

All Rsif Scholars' Publications

Accurate detection of abnormal hemoglobin variations is paramount for early diagnosis of sickle cell disease (SCD) in newborns. Traditional methods using isoelectric focusing (IEF) with agarose gels are technician-dependent and face limitations like inconsistent image quality and interpretation challenges. This study proposes a groundbreaking solution using deep learning (DL) and artificial intelligence (AI) while ensuring human guidance throughout the process. The system analyzes IEF gel images with convolutional neural networks (CNNs), achieving over 98% accuracy in identifying various SCD profiles, far surpassing the limitations of traditional methods. Furthermore, the system addresses ambiguities by incorporating an “Unconfirmed” category for unclear cases …