Development of high-throughput techniques to assess bacteriocin activity for targeted pathogen management

Introduction/Objectives: As antibiotic resistance in bacterial pathogens is on the rise, antimicrobial peptides (AMPs) offer an effective solution against potential global pandemics. These AMPs, termed ‘bacteriocins’ are naturally produced by many microbial strains. AMPs may provide an alternative strategy to clinical antibiotic use due to their diverse mechanisms of action and diminished imposition of resistance. In an effort to develop high-throughput methods to assay bacteriocin activity against known pathogens, we employed spot plate, well diffusion, and microplate reader techniques to analyze the growth of bacteriocin-producing microbes on Streptococcus agalactiae (Group B Strep) and Staphylococcus aureus (MRSA). In order to assess the collateral damage to beneficial microbes such as Lactic Acid Bacteria, we also considered the bacteriocin activity against Lactobacillus spp. Our objective was to determine prejudicial effects of various species within the Bacillus genus (Bacillus cereus, Bacillus pumilus, Bacillus thuringiensis, and Bacillus subtilis) on Group B Strep, MRSA, and Lactobacillus. By developing these high-throughput methods, we will facilitate the screening of bacteriocin activity from different clinical and environmental samples against the World Health Organization’s Top Priority Pathogen List.Methods: Isolation of Lactobacillus strains was performed from a commercial toddler probiotic via serial dilution and plating onto MRS plates. Pathogenic organisms were acquired from ATCC (GBS NCTC_8532 and Staphylococcus aureus NCTC_8181) and cultured in BHI broth at 37°C aerobically. Freshly grown strains were transferred into a 96-well plate, placed into the Alto microplate reader, and conditionally assayed to establish a working SOP. The Alto device was assessed under ambient conditions using endpoint modality, followed by aerobically at 37°C using end-point and kinetic data collection, and lastly within the anaerobic chamber in kinetic mode for 24 hours. Results from the microplate reader were transformed into growth curves. Separately, Bacillus species were isolated and stamped onto BHI agar plates. After 24 hours, spot plates were overlayed with sloppy agar (0.75% BHI and MRS agar) containing active cultures of Lactobacillus, GBS, and MRSA strains. Zones of inhibition were measured after overnight incubation. A well diffusion assay (WDA) was also performed using the cell free supernatant of Bacillus strains overlayed with MRSA and GBS in 0.75% BHI broth to determine whether bacteriocin activity is associated with cells or is released into the environment.Results: Growth curves were observed for Streptococcus agalactiae and Staphylococcus aureus in both the incubator and anaerobic chamber. Antimicrobial activity was confirmed using spot plate and WDAs. The stamped Bacillus plates overlayed with 0.75% BHI and 0.75% MRS agar with probiotics revealed zones of inhibition.Conclusion: In this study, known bacteriocin producing Bacillus species were used to determine their inhibitory effects on clinical pathogens by using augmented techniques. Further development of this high-throughput methodology will facilitate our ability to identify unknown bacteriocin-producing organisms and add to our arsenal of antimicrobial compounds

Evaluating data-sharing policies and author compliance in leading orthopedics journals

Introduction/Objectives: Orthopedic surgery is a critical field, impacting global healthcare expenditure and patient outcomes. Despite substantial research funding, issues of transparency and reproducibility persist, undermining the credibility of published findings. Data-sharing initiatives aim to address these challenges by promoting accessibility and enhancing research reliability. We aimed to assess the landscape of data-sharing practices within orthopedic surgery, focusing on the top orthopedic journals from 2020 to 2023.Methods: Original research articles from ten of the top orthopedic journals were screened and analyzed for data-sharing statements (DSS). Furthermore, we identified influential variables on the inclusion of DSS in orthopedic clinical-studies; and thematically analyzed their content to identify prevalent themes. Lastly, corresponding authors were contacted to assess their willingness to share their data.Results: Of the 1,084 articles reviewed, only 14% included DSS. The Journal of Bone and Joint Surgery demonstrated the highest proportion of articles with DSS. Over time, clinical trials exhibited an increasing trend in DSS adoption contrasting with consistently low rates among cohort studies. Thematic analysis identified gatekeeper role and conditional data availability as predominant themes within orthopedic DSS. Of the emails sent to corresponding authors, only 28 (24.35%) responded; and of those who responded only 12 (42.86%) expressed a willingness to share their data.Conclusions: Our findings underscore a significant disparity in data-sharing practices across orthopedic journals, highlighting the need for standardization and mandates for DSS. Adopting the Transparency and Openness Promotion (TOP) Guidelines can enhance accountability and foster a culture of open science within the field. By addressing these shortcomings, orthopedic journals can improve research reproducibility and advance scientific knowledge effectively

Evaluating the usefulness of randomized clinical trials in prostate cancer screening: A systematic review

Background: Prostate Cancer screening plays a crucial role in identifying the second leading cause of cancerrelated deaths worldwide, with more than 1.2 million cases reported in 2018 alone. Effective screening methods aim to achieve early detection and prompt management, while limiting overdiagnosis and treatment. Randomized controlled trials serve as the gold standard for evaluating screening methods and form the foundation for developing screening guidelines. However, there is concern in the research community about accumulating waste in clinical literature as some of these studies lack critical elements necessary for reproducibility and validity. This study aims to evaluate both the usefulness and transparency of randomized controlled trials on prostate cancer screening, utilizing a previously published 13-item usefulness criteria developed by van ’t Hooft et al. The goal is to ultimately evaluate the overall utility of the trials.Methods: This systematic review analyzes the 13 criteria in 147 RCTs on prostate cancer screening identified through PubMed and Embase published between 1994 and 2024. Articles were screened in a double blind, masked fashion by two examiners, with discrepancies resolved by a third examiner. The usefulness criteria that were assessed were scored on a point system, with 0-2 points available for each item leading to a total possible score of 26. Statistical analysis included descriptive statistics on the number and proportion of criteria met, along with linear regression plots comparing the sum of the satisfied usefulness criteria over time with the sum of transparency criteria met over time.Results: Ninety-five percent of studies scored within the lower two thirds of the total 26-point scale, with a mean score of 10.3. Criteria such as problem-base (n=144, 98%) scored highly in terms of satisfaction while key areas like patient-centeredness (n = 68, 46.3%) and information gain (n=103.0 70.1%) were largely absent in most of the studies. Regarding the transparency criteria, there was generally low satisfaction, as indicated in absence of preregistration (n=111.0, 75.5%), and publicly published protocols (n=130, 88.4%). However, overtime there is an upward trend in satisfaction of both usefulness and transparency (r = 0.21, p = 0.010) criteria with a greater positive correlation between the sum of transparency criteria overtime.Discussion: Although there are areas within these clinical trials that satisfy the usefulness criteria, significant gaps remain, contributing to literature that lacks reproducibility, generalizability or validity. Existing transparency guidelines may explain the observed upward trend of satisfaction over time. However, it has become clear that incorporating the usefulness criteria is essential to further improve the effectiveness of clinical trials and reduce waste. The application of the usefulness criteria has shown its ability to identify areas of weakness and potentially aid in the development of future guidelines during the planning phase of clinical trials.Conclusion: This study emphasizes the importance of incorporating standardized usefulness assessment tools, like the one studied, to optimize resources in clinical trial development and information reporting, ultimately improving the effectiveness of screening for life-threatening cancers.Level of Evidence: Level I

Endorsement of artificial intelligence guidelines across leading rehabilitation journals: A series of cross-sectional reviews

Background: The use of Artificial intelligence (AI) is reshaping Physical Medicine and Rehabilitation research by enabling advancements in data analysis, systematic reviews, and clinical applications. However, its adoption brings forth issues regarding ethical use, transparency, and reproducibility. This study investigates how top journals in Physical Medicine and Rehabilitation address these challenges and opportunities through their author instructions and policies.Methods: A cross-sectional review was conducted on the leading 100 peer-reviewed Journals in Physical Medicine and Rehabilitation, as ranked by the 2023 SCImago SJR indicator. Data were extracted from each journal’s “Instructions for Authors” to assess AI-related policies, including AI-specific reporting guidelines, authorship criteria, and the use of AI in manuscript preparation and image generation. Correlational analyses were conducted to explore the relationship between AI policies and journal characteristics.Results: Of the 100 journals reviewed, 86% addressed the use of AI in their instructions, with the majority prohibiting AI authorship and requiring disclosure of AI involvement in submissions. However, AI-generated content was permitted by 61% of journals, while 39 % approved the use of AI-generated images. Journals with higher impact factors were more likely to include detailed AI policies, though notable gaps in guidance and standardization remain.Conclusion: Although many Physical Medicine and Rehabilitation journals acknowledge the role of AI in research, only a few have adopted AI-specific reporting guidelines, limiting standardization and transparency of AI usage. We recommend implementing comprehensive guidelines to promote ethical, reproducible, and high-quality research in the era of AI-driven innovation

State of data sharing in plastic surgery: A cross-sectional analysis of journal practices and author adherence

Introduction: Data sharing is crucial for transparency and reproducibility in research. We aim to evaluate data sharing practices in plastic surgery research, focusing on data sharing statements (DSS), identifying key trends, and assessing follow-through.Methods: We conducted a systematic review of the top seven plastic surgery journals, selecting original research articles published from 2018 to 2023. Data extraction was performed in a masked duplicate manner, capturing DSS presence, funding sources, study design, and publication year. Trends in DSS inclusion over time, study designs, and journals were analyzed. A thematic analysis was conducted on DSS content. Corresponding authors of studies that stated data was available upon request were contacted to assess follow-through.Results: Our review included 727 articles, with only 1.51% (11/727) including DSS. DSS prevalence varied, with the highest in Aesthetic Plastic Surgery (4.42%, 5/113) and the lowest in Plastic and Reconstructive Surgery (0.43%, 1/235). Clinical trials were the most likely to include DSS (1.92%, 9/469), followed by cohort studies (1.16%, 2/173). Private funding and certain publishers were negatively associated with DSS inclusion, while journal impact factors showed a positive correlation. Responses from authors were not received, indicating a gap between stated intentions and actual practices.Conclusion: Data sharing practices in plastic surgery journals are inconsistent. Although DSS inclusion is encouraged, many studies still lack DSS, and the quality varies. Clearer mandates and enforcement are needed for effective data sharing

Exploring differential gene expression in nasal-mucus derived small extracellular vesicles: Potential biomarker for respiratory health

Introduction: Respiratory diseases, like chronic rhinosinusitis, allergic rhinitis, and asthma, pose global health concerns, affecting over 500 million people and significantly contributing to the healthcare burden. Traditional diagnostic methods often lack sensitivity and specificity, necessitating the development of novel biomarkers for early and accurate detection of specific disease conditions. Small extracellular vesicles (sEVs), nano-sized vesicles released by cells, have gained attention as carriers of molecular biomarkers due to their role in cell communication and their presence in various biological fluids. This study aims to identify respiratory-specific biomarkers within sEVs derived from nasal mucus, offering a non-invasive approach to diagnosing respiratory conditions.Methods: We received deidentified nasal mucus and serum samples from healthy individuals and extracted the sEVs using precipitation and size exclusion-based techniques. The sEVs were characterized using dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), and western blotting. We subsequently employed RNA sequencing to identify potential biomarkers within these vesicles. In addition, we performed single-cell RNA sequencing for nasal tissue to achieve a high-resolution view of cellular heterogenicity and to compare cell-type-specific biomarkers. We processed the sequencing data using bioinformatic tools including alignment, normalization and differential gene expressions to identify potential respiratory biomarkers from these health individual samples.Results: Our analysis showed a higher level of specific mRNA transcripts, associated with inflammation, cellular stress response, and membrane integrity, which are crucial for respiratory diseases in nasal sEVs compared to serum sEVs. The findings suggest that sEVs from nasal mucus are a rich source of respiratory-specific biomarkers, which could be utilized to diagnose and monitor inflammation and olfactory dysfunction in respiratory conditions.Conclusion: These sEV based biomarkers offer a promising avenue for developing non-invasive diagnostic tools that could improve patient outcomes by enabling earlier and more accurate detection of respiratory diseases. Future research should focus on validating these biomarkers in larger, diseasespecific cohorts

Effect of aging on antibody production against the receptor binding domain of Clostridioides difficile toxin B in a murine model

Clostridioides difficile is a spore producing, gram positive, anaerobic bacillus which produces toxins and causes antibiotic associated diarrhea. The main risk factor for infection is disruption of the normal gut flora from antibiotic use, allowing overgrowth of C. difficile in the colon. Toxins can cause severe diarrhea and pseudomembranous colitis. Current therapies for acute infection are Fidaxomicin, Vancomycin, or Metronidazole. C. difficile infections (CDI) are an urgent problem, and developing effective vaccines should be a priority. The current aim of vaccine development has surrounded the receptor binding domain of the toxin B protein (rTcdB) because of its increased immunogenicity and non-toxicity. CDI are frequently acquired in hospitals and nursing homes, leaving the elderly population at a higher risk of developing CDI. Additionally, immunosenescence, decreased immune functioning in older individuals, only furthers their risk. Given that most murine vaccine studies use younger mice, and the increased risk of severe infection in the elderly, it is important that the efficacy of any vaccine be tested in this population. Our lab has studied the effect of aging on IgG production against rTcdB protein in a murine model. In our pilot study, we vaccinated young (6 weeks old) and elderly (72 weeks old) C57BL/6 mice with purified recombinant protein corresponding to the receptor binding domain of C. difficile toxin B (rTcdB). Each mouse received by intraperitoneal injection of 3 doses of 20µg rTcdB mixed with Alum or saline only. Our study showed that in both young and elderly mice, rTcdB vaccination induced antigen specific IgGs in serum collected 2 weeks post vaccination. Interestingly, our novel study showed a decreased IgG response in older mice when compared to the younger group, which could indicate a decreased defense against C. difficile infection in older mice. Our study suggests that the immune system of elderly individuals may not respond as robustly against rTcdB, which could reduce the efficacy of a vaccine based on this protein. More work can be done to investigate how to support gut microbiota and reduce severity of acute CDI in at risk populations. We are currently working on exploring the role of micronutrient supplementation to improve vaccine efficiency

Two relationships, one family: Comparing parent-child bonds in immigrant families through language brokering in the family system

Intra-Family Language Brokering (IFLB) occurs when children of immigrant families translate conversations between family members (e.g. a child translates between their Spanish speaking parent and their limited Spanish speaking sibling) (Leon Cartagena et al, 2024). Often this is due to language attrition in youth cause by English only expectations at school and in the host country. The consequences of language brokering in children have been documented only in external contexts such as in the bank and stores but only two journal articles have mentioned the presence of IFLB in immigrant families. Language brokering positions children in very influential roles in families, roles that may or may not be developmentally appropriate (Tse, 1995b).parents, but. Children who serve as language brokers not only translate and interpret language for their parents but also may be expected to interpret the subtle often-subtle cultural norms that are part of the interactions of families (cite). This research investigates the relationship between parents and their translating children (language brokers or LB) and their children who need translation (NLB). Through qualitative interviews with Spanish-speaking parents and both of their children (NLB and LB) we explore how these distinct roles influence emotional bonds, communication patterns, and family dynamics. These findings reveal that the translating child (LB) often assumes increased responsibility and emotional overload leading to role reversal, however it can also lead to a stronger bond with their parent. In contrast, the child who needs translation (NLB) may experience a sense of distance from the parent due to the language barriers between them potentially affecting emotional closeness and parental authority. The study highlights the complex interplay between language, culture, and family relationships, offering insights into how translation roles shape sibling dynamics and parental connections. By comparing these two relationships, this research underscores the importance of supporting bilingualism and fostering equitable family communication to strengthen bonds across generations. The poster presentation will visually illustrate these comparisons, emphasizing the emotional and relational consequences of language roles within the family unit.Oklahoma Louis Stokes Alliance for Minority Participation ProgramEducation and Human Science

Effects of pinto bean and resistant starch supplementation on memory and cognition in estrogen-deficient mice

In the U.S., ~70% of Alzheimer’s disease (AD) cases are women. AD pathology progression and memory impairment are more severe in women, which could be triggered by estrogen loss during menopause. Three-month-old C57BL/6 female mice were injected daily with vinyl cyclohexene diepoxide (VCD), an estrogen depletion drug, or vehicle (VH) for 30 days. Mice were then randomly assigned to one of three dietary groups (n=10/group): control, 10% pinto bean (PB), and resistant starch (RS3) for 12 weeks. Behavioral tasks were then conducted, including open-field (OF) maze (a measure of anxiety/stress and exploration), Y-maze (a measure of short-term memory and exploration), and the Morris water maze (MWM) (a measure of learning ability, short/long-term memory, and cognitive function). At harvest, fecal short-chain fatty acids (SCFAs) and β-glucuronidase were quantified. The OF trials showed no differences between the VCD and VH mice. MWM 24-hour probe trials showed no differences between estrogen status nor diet. However, in the MWM 72-hour probe trials, VH mice traveled further in the annular zone than VCD mice, but differences between dietary groups are unclear. In the Y-Maze, PB-VH mice spent more time mobile than PB-VCD and Control-VH mice and traveled further in the novel arm than PB-VCD mice. Fecal SCFAs and β-glucuronidase were measured at harvest, with RS3 dietary group demonstrating significant elevation of both parameters. The PB dietary group demonstrated higher fecal SCFAs, but not β-glucuronidase. In conclusion, behavior data indicate that VCD mice overall may tend to have lower performance in long-term memory trials than VH mice, but the overall effect of diet on cognition remains unclear. Fecal data indicate that PB and/or RS3 supplementation can significantly increase SCFA and/or β-glucuronidase, which may have benefits to neuronal health. Additional work will examine differences in progressive learning ability/strategy and differences in estrogen expression in the hippocampus to better analyze the neuronal effects of estrogen status and dietary treatment.Oklahoma State University. Niblack Research Scholars ProgramIntegrative Biolog

Homemade Food Freedom Act: House Bill No. 1032 - Replaces the Home Bakery Act of 2013 and 2017

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