Comprehensive microRNA profiling in B-cells of human centenarians by massively parallel sequencing

Background: MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression and play a critical role in development, homeostasis, and disease. Despite their demonstrated roles in age-associated pathologies, little is known about the role of miRNAs in human aging and longevity.Results: We employed massively parallel sequencing technology to identify miRNAs expressed in B-cells from Ashkenazi Jewish centenarians, i.e., those living to a hundred and a human model of exceptional longevity, and younger controls without a family history of longevity. With data from 26.7 million read

Evaluation of GPT-3.5 and GPT-4 for supporting real-world information needs in healthcare delivery

Despite growing interest in using large language models (LLMs) in healthcare, current explorations do not assess the real-world utility and safety of LLMs in clinical settings. Our objective was to determine whether two LLMs can serve information needs submitted by physicians as questions to an informatics consultation service in a safe and concordant manner. Sixty six questions from an informatics consult service were submitted to GPT-3.5 and GPT-4 via simple prompts. 12 physicians assessed the LLM responses' possibility of patient harm and concordance with existing reports from an informatics consultation service. Physician assessments were summarized based on majority vote. For no questions did a majority of physicians deem either LLM response as harmful. For GPT-3.5, responses to 8 questions were concordant with the informatics consult report, 20 discordant, and 9 were unable to be assessed. There were 29 responses with no majority on "Agree", "Disagree", and "Unable to assess". For GPT-4, responses to 13 questions were concordant, 15 discordant, and 3 were unable to be assessed. There were 35 responses with no majority. Responses from both LLMs were largely devoid of overt harm, but less than 20% of the responses agreed with an answer from an informatics consultation service, responses contained hallucinated references, and physicians were divided on what constitutes harm. These results suggest that while general purpose LLMs are able to provide safe and credible responses, they often do not meet the specific information need of a given question. A definitive evaluation of the usefulness of LLMs in healthcare settings will likely require additional research on prompt engineering, calibration, and custom-tailoring of general purpose models.Comment: 27 pages including supplemental informatio

Implementation of automated calculation of free and bioavailable testosterone in Epic Beaker laboratory information system

Background: Automated calculations by laboratory information system (LIS) are efficient and accurate ways of providing calculated laboratory test results. Due to the lack of established advanced mathematical functions and equation logic in LIS software, calculations beyond simple arithmetic functions require a tedious workaround. Free and bioavailable testosterone (BT) calculations require a quadratic solver currently unavailable as ready to use the function on most commercial LIS platforms. We aimed to develop a module within the Epic Beaker LIS to enable automatic quadratic equation solving capability and real-time reporting of calculated free and BT values. Materials and Methods: We developed and implemented an advanced calculation module from the ground up using existing basic calculation programming functions in the Epic Beaker LIS. A set of calculation variables were created, and mathematical logic and functions were used to link the variables and perform the actual quadratic equation based calculations. Calculations were performed in real-time during result entry events, and calculated results populated the result components in LIS automatically. Results: Free and BT were calculated using instrument measured results of total testosterone, sex hormone binding globulin, and/or serum albumin, by applying equations widely adopted in laboratory medicine for endocrine diseases and disorders. Calculated results in Epic Beaker LIS were then compared and confirmed by manual calculations using Microsoft Excel spreadsheets and scientific calculators to have no discrepancies. Conclusions: Automated calculations of free and BT were successfully implemented and validated, the first of such implementation for the Epic Beaker LIS platform, eliminating the need of offline manual calculations, potential transcription error, and with improved turnaround time. It may serve as a model to build similarly complex equations when the clinical need arises

Epigenetics Decouples Mutational from Environmental Robustness. Did It Also Facilitate Multicellularity?

<div><p>The evolution of ever increasing complex life forms has required innovations at the molecular level in order to overcome existing barriers. For example, evolving processes for cell differentiation, such as epigenetic mechanisms, facilitated the transition to multicellularity. At the same time, studies using gene regulatory network models, and corroborated in single-celled model organisms, have shown that mutational robustness and environmental robustness are correlated. Such correlation may constitute a barrier to the evolution of multicellularity since cell differentiation requires sensitivity to cues in the internal environment during development. To investigate how this barrier might be overcome, we used a gene regulatory network model which includes epigenetic control based on the mechanism of histone modification via Polycomb Group Proteins, which evolved in tandem with the transition to multicellularity. Incorporating the Polycomb mechanism allowed decoupling of mutational and environmental robustness, thus allowing the system to be simultaneously robust to mutations while increasing sensitivity to the environment. In turn, this decoupling facilitated cell differentiation which we tested by evaluating the capacity of the system for producing novel output states in response to altered initial conditions. In the absence of the Polycomb mechanism, the system was frequently incapable of adding new states, whereas with the Polycomb mechanism successful addition of new states was nearly certain. The Polycomb mechanism, which dynamically reshapes the network structure during development as a function of expression dynamics, decouples mutational and environmental robustness, thus providing a necessary step in the evolution of multicellularity.</p></div

Polycomb functionality promotes the creation of multiple input/output mappings for the same network.

<p>In order to determine if a certain expression thresholds are required for the ability to add new input/output mappings we tested the Polycomb threshold (γ) over a wide range of values. When a threshold of 0 is selected no genes are ever be repressed by polycomb (the same as the default cis-regulatory case). At a threshold of 1.0 every gene with a PRE would be repressed, effectively reducing the number of genes in the network. We see the threshold of the Polycomb is not essential for the ability to add new states just the general functionality, N = 200. Inset: With all parameters remaining constant the introduction of polycomb functionality allows the system to add states 99.5% of the time where-as the standard cis-regulatory network is only able to add states 53% of the time. (n = 200, p = 8.62×10⁻²²).</p

Polycomb functionality decouples environmental and mutational robustness.

<p>A gene regulatory network evolving under stabilizing selection without Polycomb, both mutational and environmental robustness increase together over time (blue). With Polycomb the system shows reduced environmental robustness while development of mutational robustness continues (red). All data shown are averages over 200 independent trials using a randomly selected founder individual; error bars represent the SEM, ,  = 0.05,  = 2 (see <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1003450#s4" target="_blank">Methods</a>).</p

Effective network stratifications by environmental conditions via Polycomb.

<p>A) The Polycomb mechanism allows cells to have drastically different phenotypes even though the genotype is identical. Activation of Polycomb during development forces Polycomb-susceptible genes with low levels of expression to be permanently repressed, via chromatin modification, effectively trimming the network (faded interactions). (B and C) Different environmental conditions allow a single network to be trimmed effectively into different networks distinct output states. The final possible outputs of the networks are intimately tied to the original environmental inputs.</p

Estimating the efficacy of symptom-based screening for COVID-19

Abstract There is substantial interest in using presenting symptoms to prioritize testing for COVID-19 and establish symptom-based surveillance. However, little is currently known about the specificity of COVID-19 symptoms. To assess the feasibility of symptom-based screening for COVID-19, we used data from tests for common respiratory viruses and SARS-CoV-2 in our health system to measure the ability to correctly classify virus test results based on presenting symptoms. Based on these results, symptom-based screening may not be an effective strategy to identify individuals who should be tested for SARS-CoV-2 infection or to obtain a leading indicator of new COVID-19 cases