Efficient electroporation of neuronal cells using synthetic oligonucleotides: identifying duplex RNA and antisense oligonucleotide activators of human frataxin expression.

Oligonucleotide drugs are experiencing greater success in the clinic, encouraging the initiation of new projects. Resources are insufficient to develop every potentially important project, and persuasive experimental data using cell lines close to disease target tissue is needed to prioritize candidates. Friedreich's ataxia (FRDA) is a devastating and currently incurable disease caused by insufficient expression of the enzyme frataxin (FXN). We have previously shown that synthetic nucleic acids can activate FXN expression in human patient-derived fibroblast cells. We chose to further test these compounds in induced pluripotent stem cell-derived neuronal progenitor cells (iPSC-NPCs). Here we describe methods to deliver oligonucleotides and duplex RNAs into iPSC-NPCs using electroporation. Activation of FXN expression is potent, easily reproducible, and potencies parallel those determined using patient-derived fibroblast cells. A duplex RNA and several antisense oligonucleotides (ASOs) with different combinations of 2'-methoxyethyl (2'-MOE), 2'-fluoro (2'-F), and constrained ethyl (cEt) were active, providing multiple starting points for further development and highlighting improved potency as an important goal for preclinical development. Our data support the conclusion that ASO-mediated activation of FXN is a feasible approach for treating FRDA and that electroporation is a robust method for introducing ASOs to modulate gene expressions in neuronal cells.

International Nursing Program Accreditation.

The Accreditation Commission for Education in Nursing (ACEN) is committed to being a supportive partner in strengthening the quality of nursing education for all levels of nursing programs domestically and internationally. With a longstanding history of accreditation dating back 66 years, the ACEN accredited its first international program in 2004 adding international accreditation to its repertoire. Recognizing geographic, cultural, and national differences, the ACEN common core of Standards and Criteria equip faculty with autonomy to embrace unique attributes of their programs regardless of location, culture, and nationality. Further, the ACEN review process fosters self-evaluation, peer review, and the promotion of educational equity, access, and mobility. As a result, the number of international nursing programs pursuing and attaining accreditation with the ACEN has increased thus validating the inclusiveness and relevance of the ACEN Standards and Criteria. The purpose of this article is to highlight ways in which ACEN Standards and Criteria apply to domestic and international nursing programs.

ACEN accreditation perspectives: Distance education.

Many nursing programs implemented the use of distance education in response to the COVID-19 pandemic. Faculty must consider the mission of the governing organization and the fiscal, physical, technological, and learning resources available while ensuring the quality of the education being provided when using any type of distance learning method of delivery. The purpose of this article is to discuss how selected 2017 ACEN Accreditation Standards and Criteria can assist the faculty in ensuring quality nursing education in distance education methods of delivery.

Development of the Doctor of Nursing Practice Nurse Practitioner Minimum Data Set (DNP NP MDS).

BACKGROUND: While the number of Doctor of Nursing Practice (DNP) programs has grown steadily, there is limited data on how national organizations are collecting data on DNP-prepared nurse practitioners (NPs) and no standard instrument exists to collect data on DNP-prepared NPs. PURPOSE: The purpose of this study was to develop a universal minimum data set (MDS) for the DNP-prepared NP population. METHOD: Instrument development consisted of several sequential stages, including conceptualization and item generation, preliminary evaluation of items, field testing the survey, and analysis of scale development data. FINDINGS: A set of 16 core variables and 19 additional variables were developed to collect standardized data on the demographics, education, and practice patterns of DNP-prepared NPs. Pilot testing revealed high correlations between the activities DNP-prepared NPs are prepared for and typically participate in a typical workweek and in their career. The MDS demonstrated high reliability in our sample. DISCUSSION: The DNP NP MDS can be used for data collection by various stakeholders, including national organizations, to facilitate improved tracking of outcome data for the DNP-prepared NP workforce. It can also provide data-driven support for the need and significance of the DNP degree for NPs.

Responding to the pandemic: Nursing education and the ACEN.

The purpose of this article is to provide an overview of some of the ways in which nursing education programs and the Accreditation Commission for Education in Nursing (ACEN) adapted to the challenges of the COVID-19 pandemic. Although the longer-term impacts of the pandemic cannot be known, it is clear that nurse educators rose to the challenge of educating nurses during a pandemic through rapid implementation of innovative teaching strategies and student support. The article also provides some guidance for ACEN-accredited nursing education programs regarding substantive changes that may be permanently implemented as a result of what programs learned during the pandemic.

Analytical validation of a novel multiplex test for detection of advanced adenoma and colorectal cancer in symptomatic patients.

Early detection of colorectal cancer (CRC) is key to reducing associated mortality. Despite the importance of early detection, approximately 40% of individuals in the United States between the ages of 50-75 have never been screened for CRC. The low compliance with colonoscopy and fecal-based screening may be addressed with a non-invasive alternative such as a blood-based test. We describe here the analytical validation of a multiplexed blood-based assay that measures the plasma concentrations of 15 proteins to assess advanced adenoma (AA) and CRC risk in symptomatic patients. The test was developed on an electrochemiluminescent immunoassay platform employing four multi-marker panels, to be implemented in the clinic as a laboratory developed test (LDT). Under the Clinical Laboratory Improvement Amendments (CLIA) and College of American Pathologists (CAP) regulations, a United States-based clinical laboratory utilizing an LDT must establish performance characteristics relating to analytical validity prior to releasing patient test results. This report describes a series of studies demonstrating the precision, accuracy, analytical sensitivity, and analytical specificity for each of the 15 assays, as required by CLIA/CAP. In addition, the report describes studies characterizing each of the assays' dynamic range, parallelism, tolerance to common interfering substances, spike recovery, and stability to sample freeze-thaw cycles. Upon completion of the analytical characterization, a clinical accuracy study was performed to evaluate concordance of AA and CRC classifier model calls using the analytical method intended for use in the clinic. Of 434 symptomatic patient samples tested, the percent agreement with original CRC and AA calls was 87% and 92% respectively. All studies followed CLSI guidelines and met the regulatory requirements for implementation of a new LDT. The results provide the analytical evidence to support the implementation of the novel multi-marker test as a clinical test for evaluating CRC and AA risk in symptomatic individuals.

Detection and Characterization of Circulating Tumour Cells in Multiple Myeloma.

Multiple myeloma (MM) remains an incurable disease despite recent therapeutic improvements. The ability to detect and characterize MM circulating tumour cells (CTCs) in peripheral blood provides an alternative to replace or augment invasive bone marrow (BM) biopsies with a simple blood draw, providing real-time, clinically relevant information leading to improved disease management and therapy selection. Here we have developed and qualified an enrichment-free, cell-based immunofluorescence MM CTC assay that utilizes an automated digital pathology algorithm to distinguish MM CTCs from white blood cells (WBCs) on the basis of CD138 and CD45 expression levels, as well as a number of morphological parameters. These MM CTCs were further characterized for expression of phospho-ribosomal protein S6 (pS6) as a readout for PI3K/AKT pathway activation. Clinical feasibility of the assay was established by testing blood samples from a small cohort of patients, where we detected populations of both CD138pos and CD138neg MM CTCs. In this study, we developed an immunofluorescent cell-based assay to detect and characterize CTCs in MM.

Detection and Characterization of Circulating Tumour Cells from Frozen Peripheral Blood Mononuclear Cells.

Retrospective analysis of patient tumour samples is a cornerstone of clinical research. CTC biomarker characterization offers a non-invasive method to analyse patient samples. However, current CTC technologies require prospective blood collection, thereby reducing the ability to utilize archived clinical cohorts with long-term outcome data. We sought to investigate CTC recovery from frozen, archived patient PBMC pellets. Matched samples from both mCRPC patients and mock samples, which were prepared by spiking healthy donor blood with cultured prostate cancer cell line cells, were processed "fresh" via Epic CTC Platform or from "frozen" PBMC pellets. Samples were analysed for CTC enumeration and biomarker characterization via immunofluorescent (IF) biomarkers, fluorescence in-situ hybridization (FISH) and CTC morphology. In the frozen patient PMBC samples, the median CTC recovery was 18%, compared to the freshly processed blood. However, abundance and localization of cytokeratin (CK) and androgen receptor (AR) protein, as measured by IF, were largely concordant between the fresh and frozen CTCs. Furthermore, a FISH analysis of PTEN loss showed high concordance in fresh vs. frozen. The observed data indicate that CTC biomarker characterization from frozen archival samples is feasible and representative of prospectively collected samples.