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Abstract Biomedicine today is experiencing a shift towards decentralized data collection, which promises enhanced reproducibility and collaboration across diverse laboratory environments. This inter-laboratory study evaluates the performance of biocytometry, a method utilizing engineered bioparticles for enumerating cells based on their surface antigen patterns. In a decentralized framework, spanning 78 assays conducted by 30 users across 12 distinct laboratories, biocytometry consistently demonstrated significant statistical power in discriminating numbers of target cells at varying concentrations as low as 1 cell per 100,000 background cells. User skill levels varied from expert to beginner capturing a range of proficiencies. Measurement was performed in a decentralized environment without any instrument cross-calibration or advanced user training outside of a basic instruction manual. The results affirm biocytometry to be a viable solution for immunophenotyping applications demanding sensitivity as well as scalability and reproducibility and paves the way for decentralized analysis of rare cells in heterogeneous samples. 

MicroRNAs are small noncoding nucleotides that serve as intracellular and extracellular signaling molecules. A previous collaboration found miR-127/3p circulation in the blood of breast cancer patients correlated with improved patient recovery and prognosis. While this study exclusively focused on breast cancer patients, data mining of the TCGA databases indicated that miR-127/3p may be positively associated with outcomes in other cancer types. In our study, A549 lung adenocarcinoma cells were transfected with miR-127/3p using Cell Block protocols produced by the Cell Biology Education Consortium (CBEC). After transfection, cell migration (scratch/wound healing) assays were used to determine the role miR-127/3p plays in the tumor microenvironment. To mimic and test this environment, transfected cells were incubated in normal oxygen (normoxic) and low oxygen (hypoxic) environments. We found that miR-127/3p inhibited cell migration in both normal oxygen and hypoxic environments. These results help elucidate the role miR-127/3p plays in the prevention of metastasis and further highlight its potential as a positive biomarker. 

There is a growing need for the development and communication of cell culture-based laboratory activities specifically designed for undergraduate students. This multi-week laboratory activity allows students to take part in the planning, experimentation, data analysis, and communication of the results of their cell culture-based research project. The laboratory activity specifically uses fatty acid induction of lipid droplets in cancer HeLa cells followed by a novel live-cell staining protocol developed specifically to allow undergraduate students the opportunity to complete a cell culture-based fluorescence microscopy project. This laboratory activity incorporates multiple levels of assessment and allows students to explore the responses of HeLa cancer cells to their environment. 

ABSTRACT During the COVID-19 pandemic, biology educators were forced to think of ways to communicate with their students, engaging them in science and with the scientific community. For educators using course-based undergraduate research experiences (CUREs), the challenge to have students perform real science, analyze their work, and present their results to a larger scientific audience was difficult as the world moved online. Many instructors were able to adapt CUREs utilizing online data analysis and virtual meeting software for class discussions and synchronous learning. However, interaction with the larger scientific community, an integral component of making science relevant for students and allowing them to network with other young scientists and experts in their fields, was still missing. Even before COVID-19, a subset of students would travel to regional or national meetings to present their work, but most did not have these opportunities. With over 300 million active users, Twitter provided a unique platform for students to present their work to a large and varied audience. The Cell Biology Education Consortium hosted an innovative scientific poster session entirely on Twitter to engage undergraduate researchers with one another and with the much broader community. The format for posting on this popular social media platform challenged students to simplify their science and make their points using only a few words and slides. Nineteen institutions and over one hundred students participated in this event. Even though these practices emerged as a necessity during the COVID-19 pandemic, the Twitter presentation strategy shared in this paper can be used widely. 

Background: Neoadjuvant chemotherapy (NACT) is an increasingly used approach for treatment of breast cancer. The pathological complete response (pCR) is considered a good predictor of disease-specific survival. This study investigated whether circulating exosomal microRNAs could predict pCR in breast cancer patients treated with NACT. Method: Plasma samples of 20 breast cancer patients treated with NACT were collected prior to and after the first cycle. RNA sequencing was used to determine microRNA profiling. The Cancer Genome Atlas (TCGA) was used to explore the expression patterns and survivability of the candidate miRNAs, and their potential targets based on the expression levels and copy number variation (CNV) data. Results: Three miRNAs before that NACT (miR-30b, miR-328 and miR-423) predicted pCR in all of the analyzed samples. Upregulation of miR-127 correlated with pCR in triple-negative breast cancer (TNBC). After the first NACT dose, pCR was predicted by exo-miR-141, while miR-34a, exo-miR182, and exo-miR-183 predicted non-pCR. A significant correlation between the candidate miRNAs and the overall survival, subtype, and metastasis in breast cancer, suggesting their potential role as predictive biomarkers of pCR. Conclusions: If the miRNAs identified in this study are validated in a large cohort of patients, they might serve as predictive non-invasive liquid biopsy biomarkers for monitoring pCR to NACT in breast cancer. 

ABSTRACT Course-based undergraduate research experiences (CUREs) provide a way for students to gain research experience in a classroom setting. Few examples of cell culture CUREs or online CUREs exist in the literature. The Cell Biology Education Consortium (CBEC) provides a network and resources for instructors working to incorporate cell-culture based research into the classroom. In this article, we provide examples from six instructors from the CBEC network on how they structure their cell-culture CUREs and how they transitioned the labs to online in the spring semester of 2020. We intend for these examples to provide instructors with ideas for strategies to set up cell culture CUREs, how to change that design mid-term, and for creating online CUREs in the future. 

At institutions with an emphasis on authentic research experiences as an integral part of the biology curriculum, COVID created a huge challenge for course instructors whose learning objectives were designed for such experiences. Moving such laboratory experiences online when remote learning became necessary has resulted in a new model for CUREs that utilizes free online databases to provide not only a novel research experience for students, but also the opportunity to engage in big data analysis. Cancer BioPortal (cBioPortal) is an open-access collective cancer research resource for storing and exploring clinical, genomic, proteomic, and transcriptomic data. cBioPortal eliminates the computational barrier of interpreting complex genomic data by providing easily understandable visualization that can be interpreted and translated into relevant biological insights. Because no prior computational knowledge is required, cBioPortal is an ideal educational tool for either in-person or distance learning environments. We developed a pedagogical approach, video tutorials, and data analysis workflows centered on using cBioPortal. Pedagogically, students develop an initial research outline that is continually updated and graded throughout the project. Progress during the project or course is assessed by a series of student presentations that are 5 to 15 minutes in length and are aimed at explaining the approach used in data acquisition, interpretation of the data, and relevance to the initial hypothesis. While cancer-specific, this analysis platform appeals to a wide range of classes and student interests. Further, the project has been successfully done both as an independent research experience and as part of a virtual class-based research project. 

Exosomes are extracellular vesicles ranging in size from 40 to 100nm that are used for extracellular communication to control the cell niche environment with regards to differentiation. Initial studies in our lab have shown that exosomes isolated from differentiating neurites can cause differentiation in cells absent typical neuronal hormones. However, these studies only used exosomes isolated after complete differentiation. To further clarify the role of exosomes in cell differentiation, we isolated exosomes from NGF-treated PC12 cells after two, four and six days. The exosomes were taken at the specific times due to a slight majority of cells showing differentiation at day three, with an overwhelming majority showing differentiation at day six. Day two, four and six were thus good markers in the development of cell differentiation. Each set of exosomes was added to a different well of untreated cells, which were then observed for differentiation daily over a six-day period. The results showed a considerably larger amount of percent differentiation in cells treated with the day six exosomes compared to cells treated by the day two and day four exosomes. The change in number of differentiated cells in different exosome treatments indicate that exosomal contents change over time. In an effort to make sure that no NGF contamination occurred in our treatments, an NGF-ELISA was completed testing all the treatment exosomes with results showing no NGF. Additionally, exosomes were isolated from NGF spiked media to show that no NGF was carried over in our exosome isolation process. Both tests further evidence that our exosomes did not contain NGF. Our overall results indicate that the isolated exosomes changed contents over time and that their contents caused greater percent differentiation over time. Funding provided by the Cell Biology Education Consortium (CBEC) and through the AR-EPSCoR Center for Advanced Surface Engineering.