Microfluidic Squeezing Enables MHC Class I Antigen Presentation by Diverse Immune Cells to Elicit CD8+ T Cell Responses with Antitumor Activity.

CD8+ T cell responses are the foundation of the recent clinical success of immunotherapy in oncologic indications. Although checkpoint inhibitors have enhanced the activity of existing CD8+ T cell responses, therapeutic approaches to generate Ag-specific CD8+ T cell responses have had limited success. Here, we demonstrate that cytosolic delivery of Ag through microfluidic squeezing enables MHC class I presentation to CD8+ T cells by diverse cell types. In murine dendritic cells (DCs), squeezed DCs were ∼1000-fold more potent at eliciting CD8+ T cell responses than DCs cross-presenting the same amount of protein Ag. The approach also enabled engineering of less conventional APCs, such as T cells, for effective priming of CD8+ T cells in vitro and in vivo. Mixtures of immune cells, such as murine splenocytes, also elicited CD8+ T cell responses in vivo when squeezed with Ag. We demonstrate that squeezing enables effective MHC class I presentation by human DCs, T cells, B cells, and PBMCs and that, in clinical scale formats, the system can squeeze up to 2 billion cells per minute. Using the human papillomavirus 16 (HPV16) murine model, TC-1, we demonstrate that squeezed B cells, T cells, and unfractionated splenocytes elicit antitumor immunity and correlate with an influx of HPV-specific CD8+ T cells such that >80% of CD8s in the tumor were HPV specific. Together, these findings demonstrate the potential of cytosolic Ag delivery to drive robust CD8+ T cell responses and illustrate the potential for an autologous cell-based vaccine with minimal turnaround time for patients.

Outcomes of patients in Chagas disease of the central nervous system: a systematic review.

Chagas disease is a parasitic infection caused by the protozoan Trypanosoma cruzi. One of the complications of the disease is the infection of the central nervous system (CNS), as it can result from either the acute phase or by reactivation during the chronic phase, exhibiting high mortality in immunocompromised patients. This systematic review aimed to determine clinical and paraclinical characteristics of patients with Chagas disease in the CNS. Articles were searched from PubMed, Scopus and LILACS until January 2023. From 2325 articles, 59 case reports and 13 case series of patients with Chagas in the CNS were retrieved from which 138 patients were identified. In this population, 77% of the patients were male, with a median age of 35 years old, from which most of them came from Argentina and Brazil. Most of the individuals were immunocompromised from which 89% were HIV-positive, and 54 patients had an average of 48 cells per mm3 CD4+ T cells. Motor deficits and seizures were the most common manifestation of CNS compromise. Furthermore, 90 patients had a documented CNS lesion by imaging from which 89% were supratentorial and 86% were in the anterior/middle cranial fossa. The overall mortality was of 74%. Among patients who were empirically treated with anti-toxoplasma drugs, 70% died. This review shows how Chagas disease in the CNS is a devastating complication requiring prompt diagnosis and treatment to improve patients' outcomes.

PAs and NPs in employee health during the COVID-19 pandemic.

ABSTRACT: COVID-19 created unprecedented occupational health challenges for hospitals. To meet these demands at a large county safety-net hospital, a COVID-19 employee response team led by PAs and NPs was created. From April 2020 through February 2022, this team managed more than 14,000 discrete employee contacts related to COVID-19 employee concerns. This article describes our experience in creating this team and highlights key strengths and lessons for other institutions seeking to adopt similar models.

False positive serology of prepandemic chagasic samples with SARS-CoV-2 antigen.

OBJECTIVE: To determine whether prepandemic sera from patients with Chagas disease recognise SARS-CoV-2 antigens. MATERIALS AND METHODS: Forty sera from patients with Chagas disease were tested for the presence of IgG cross-reactivity against the nucleocapsid protein (NP) and spike (S) SARS-CoV-2 proteins by ELISA. Positive samples were tested again using a different ELISA and CLIA, both against NP. RESULTS: None of the sera from patients with Chagas disease, previously confirmed as positive for the presence of anti-Trypanosoma cruzi antibodies reacted against the SARS-CoV-2 S protein, and six samples tested positive for the NP antigen (15%). The six positive samples were re-tested, five remained positive by ELISA and all were negative by CLIA. CONCLUSION: According to our data, false-positive results might be a concern in the detection of SARS-CoV-2 antibodies in patients with Chagas disease.

Suicide risk and prevention in LGBTQ+ youth.

ABSTRACT: Lesbian, gay, bisexual, transgender, questioning (or queer), and other sexual and gender minority (LGBTQ+) youth face various forms of bullying and mistreatment that may lead to suicide. Nurses can help recognize, address, and mitigate challenges associated with suicide among LGBTQ+ youth.

Diminished mitogen-induced T cell proliferation by Trypanosoma cruzi antigens associated with antigen-presenting cell modulation and CD3 signaling.

Chronic infection by Trypanosoma cruzi decreases T cell proliferation and it is most likely accompanied by changes in signals required for activation. We assessed the effect of T. cruzi antigens on mitogen-induced proliferation of T cells from uninfected individuals and the association with the expression of molecules involved in antigen presentation, T cell costimulation and activation, and cytokine production. T. cruzi antigen exposure reduced mitogen-induced proliferation of CD4+ and CD8+ T cells in PBMC cultures, but only reduced mitogen-induced proliferation in the CD4+ T cells from sorted cell cultures cocultured with antigen-pulsed CD3- cells. CD40/CD80 and CD86 expression were reduced in antigen-pulsed DCs and monocytes, respectively. TNF-α, IL-10 and CCL17 levels were increased in cultures with antigen-pulsed CD3- cells, while CD3ζ chain expression was reduced in T cells from cultures with antigen. Our findings suggest that T. cruzi could alter T cell proliferation indirectly by downregulating costimulatory molecules and inducing the secretion of IL-10 and directly by decreasing TCR signaling.

Biallelic KARS pathogenic variants cause an early-onset progressive leukodystrophy.

The leukodystrophies cause severe neurodevelopmental defects from birth and follow an incurable and progressive course that often leads to premature death. It has recently been reported that abnormalities in aminoacyl t-RNA synthetase (ARS) genes are linked to various unique leukodystrophies and leukoencephalopathies. Aminoacyl t-RNA synthetase proteins are fundamentally known as the first enzymes of translation, catalysing the conjugation of amino acids to cognate tRNAs for protein synthesis. It is known that certain aminoacyl t-RNA synthetase have multiple non-canonical roles in both transcription and translation, and their disruption results in varied and complicated phenotypes. We clinically and genetically studied seven patients (six male and one female; aged 2 to 12 years) from five unrelated families who all showed the same phenotypes of severe developmental delay or arrest (7/7), hypotonia (6/7), deafness (7/7) and inability to speak (6/7). The subjects further developed intractable epilepsy (7/7) and nystagmus (6/6) with increasing age. They demonstrated characteristic laboratory data, including increased lactate and/or pyruvate levels (7/7), and imaging findings (7/7), including calcification and abnormal signals in the white matter and pathological involvement (2/2) of the corticospinal tracts. Through whole-exome sequencing, we discovered genetic abnormalities in lysyl-tRNA synthetase (KARS). All patients harboured the variant [c.1786C>T, p.Leu596Phe] KARS isoform 1 ([c.1702C>T, p.Leu568Phe] of KARS isoform 2) either in the homozygous state or compound heterozygous state with the following KARS variants, [c.879+1G>A; c.1786C>T, p.Glu252_Glu293del; p.Leu596Phe] ([c.795+1G>A; c.1702C>T, p.Glu224_Glu255del; p.Leu568Phe]) and [c.650G>A; c.1786C>T, p.Gly217Asp; p.Leu596Phe] ([c.566G>A; c.1702C>T, p.Gly189Asp; p.Leu568Phe]). Moreover, similarly disrupted lysyl-tRNA synthetase (LysRS) proteins showed reduced enzymatic activities and abnormal CNSs in Xenopus embryos. Additionally, LysRS acts as a non-canonical inducer of the immune response and has transcriptional activity. We speculated that the complex functions of the abnormal LysRS proteins led to the severe phenotypes in our patients. These KARS pathological variants are novel, including the variant [c.1786C>T; p.Leu596Phe] (c.1702C>T; p.Leu568Phe) shared by all patients in the homozygous or compound-heterozygous state. This common position may play an important role in the development of severe progressive leukodystrophy. Further research is warranted to further elucidate this relationship and to investigate how specific mutated LysRS proteins function to understand the broad spectrum of KARS-related diseases.

mRNA Delivery for Therapeutic Anti-HER2 Antibody Expression In Vivo.

Antibody-based drugs are a leading class of biologics used to treat a variety of diseases, including cancer. However, wide antibody implementation is hindered by manufacturing challenges and high production cost. Use of in-vitro-transcribed mRNA (IVT-mRNA) for endogenous protein expression has the potential to circumvent many of the shortcomings of antibody production and therapeutic application. Here, we describe the development of an IVT-mRNA system for in vivo delivery of a humanized anti-HER2 (also known as ERBB2) antibody, trastuzumab, and demonstrate its anticancer activity. We engineered the IVT-mRNA sequence to maximize expression, then formulated the IVT-mRNA into lipid-based nanoparticles (LNPs) to protect the mRNA from degradation and enable efficient in vivo delivery. Systemic delivery of the optimized IVT-mRNA loaded into LNPs resulted in antibody serum concentrations of 45 ± 8.6 µg/mL for 14 days after LNP injection. Further studies demonstrated an improved pharmacokinetic profile of the produced protein compared to injection of trastuzumab protein. Finally, treatment of tumor-bearing mice with trastuzumab IVT-mRNA LNPs selectively reduced the volume of HER2-positive tumors and improved animal survival. Taken together, the results of our study demonstrate that using IVT-mRNA LNPs to express full-size therapeutic antibodies in the liver can provide an effective strategy for cancer treatment and offers an alternative to protein administration.

Antiparasitic Treatment Induces an Improved CD8+ T Cell Response in Chronic Chagasic Patients.

Chagas disease is a chronic infection caused by Trypanosoma cruzi, an intracellular protozoan parasite. Chronic chagasic patients (CCPs) have dysfunctional CD8+ T cells that are characterized by impaired cytokine production, high coexpression of inhibitory receptors, and advanced cellular differentiation. Most patients diagnosed in the chronic phase of Chagas disease already exhibit heart involvement, and there is no vaccination that protects against the disease. Antiparasitic treatment is controversial as to its indication for this stage of the disease. There is a lack of biological markers to evaluate the effectiveness of antiparasitic treatment, and little is known about the effect of the treatment on CD8+ T cells. Thus, the aim of the current study was to analyze the early effects of antiparasitic treatment on CD8+ T cells from CCPs with asymptomatic clinical forms of disease. To evaluate the CD8+ T cell subsets, expression of inhibitory receptors, and functionality of T cells in CCPs, PBMCs were isolated. The results showed that treatment of CCPs with the asymptomatic form of the disease induces an increase in the frequency of CD8+ central memory T cells and terminal effector T cells, a decrease in the coexpression of inhibitory receptors, an improved Ag-specific CD8+ T cell response exhibited by the individual production of IFN-γ or IL-2, and a multifunctional CD8+ T cell profile of up to four functions (IFN-γ+IL-2+Perforin+Granzyme B+). These findings suggest that, in CCPs, antiparasitic treatment improved the quality of Ag-specific CD8+ T cell responses associated with a decrease in inhibitory receptor coexpression, which could serve as biomarkers for monitoring the effectiveness of antiparasitic treatment.

Living a Good Way of Life: Perspectives from American Indian and First Nation Young Adults.

In this study, we respond to calls for strengths-based Indigenous research by highlighting American Indian and First Nations (Anishinaabe) perspectives on wellness. We engaged with Anishinaabe community members by using an iterative, collaborative Group Concept Mapping methodology to define strengths from a within-culture lens. Participants (n = 13) shared what it means to live a good way of life/have wellness for Anishinaabe young adults, ranked/sorted their ideas, and shared their understanding of the map. Results were represented by nine clusters of wellness, which addressed aspects of self-care, self-determination, actualization, community connectedness, traditional knowledge, responsibility to family, compassionate respect toward others, enculturation, and connectedness with earth/ancestors. The clusters were interrelated, primarily in the relationship between self-care and focus on others. The results are interpreted by the authors and Anishinaabe community members though the use of the Seven Grandfather Teachings, which provide a framework for understanding Anishinaabe wellness. The Seven Grandfather Teachings include Honesty (Gwayakwaadiziwin), Respect (Manaadendamowin), Humility (Dabaadendiziwin), Love (Zaagi'idiwin), Wisdom (Nibwaakaawin), Bravery/Courage (Aakode'ewin), and Truth (Debwewin).

United States beef quality as chronicled by the National Beef Quality Audits, Beef Consumer Satisfaction Projects, and National Beef Tenderness Surveys - A review.

Meat quality is a very difficult term to define because it means different things to different people. When purchasing beef, consumers in the United States are likely to consider color, price, marbling level, subcutaneous fat trim, or cut thickness when determining the quality of beef. Once consumers have consumed the product, meat quality becomes exponentially more difficult to define due to the subjective nature of this term. Traditionally, tenderness, juiciness, and flavor have been considered the three most important factors that determine the palatability of beef. Therefore, American meat science beef research and industry focus has turned to measuring and quantifying these 3 attributes objectively and subjectively, and to determining what influences them. In reviewing the scientific literature, attempting to meaningfully summarize the findings of the thousands of studies on beef meat quality is impossible due to the inherent differences in the objective and methodology of studies. Fortunately, the United States beef industry and their national trade association, the National Cattlemen's Beef Association (NCBA), have conducted numerous surveys and audits to characterize the quality of the products being produced and marketed by their cattlemen and the palatability perceptions of their consumers. The data produced by these studies is quite large and impossible to summarize in entirety in this review. Therefore, this review concentrates on the most important attributes that determine the value of a beef carcass and objectively measured and consumer-assessed palatability characteristics of fresh meat from these carcasses from 1987 through 2010.

Reduced skeletal muscle fiber size following caloric restriction is associated with calpain-mediated proteolysis and attenuation of IGF-1 signaling.

Caloric restriction decreases skeletal muscle mass in mammals, principally due to a reduction in fiber size. The effect of suboptimal nutrient intake on skeletal muscle metabolic properties in neonatal calves was examined. The longissimus muscle (LM) was collected after a control (CON) or caloric restricted (CR) diet was cosnumed for 8 wk and muscle fiber size, gene expression, and metabolic signal transduction activity were measured. Results revealed that CR animals had smaller (P < 0.05) LM fiber cross-sectional area than CON, as expected. Western blot analysis detected equivalent amounts of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC1α) but reduced (P < 0.05) amounts of the splice-variant, PGC1α-4 in CR LM. Expression of IGF-1, a PGC1α-4 target gene, was 40% less (P < 0.05) in CR than CON. Downstream mediators of autocrine IGF-1 signaling also are attenuated in CR by comparison with CON. The amount of phosphorylated AKT1 was less (P < 0.05) in CR than CON. The ratio of p4EBP1T37/46 to total 4EBP1, a downstream mediator of AKT1, did not differ between CON and CR. By contrast, protein lysates from CR LM contained less (P < 0.05) total glycogen synthase kinase-3ß (GSK3ß) and phosphorylated GSK3ß than CON LM, suggesting blunted protein synthesis. Smaller CR LM fiber size associates with increased (P < 0.05) calpain 1 (CAPN1) activity coupled with lower (P < 0.05) expression of calpastatin, the endogenous inhibitor of CAPN1. Atrogin-1 and MuRF expression and autophagy components were unaffected by CR. Thus CR suppresses the hypertrophic PGC1α-4/IGF-1/AKT1 pathway while promoting activation of the calpain system.

Inhibitory Receptor Expression on CD8+ T Cells Is Linked to Functional Responses against Trypanosoma cruzi Antigens in Chronic Chagasic Patients.

In mammals, chronic diseases resulting from infectious agents have been associated with functional T cell response deficiency, a high frequency of terminally differentiated T cells, the presence of monofunctional Ag-specific T cells, and increased expression of inhibitory receptors. Similar to other chronic diseases, the progressive loss of certain functional activities during Trypanosoma cruzi infection might result in the inability to control replication of this parasite. To examine this hypothesis, we evaluated the differentiation and cell effector function of CD8(+) T cells and characterized the expression of inhibitory receptors and the presence of the parasite in the bloodstream of chagasic patients. The results showed that patients at an advanced severe disease stage had a higher frequency of terminally differentiated CD8(+) T cells than patients at an early stage of the disease. A monofunctional CD8(+) T cell response was observed in patients at an advanced stage, whereas the coexpression of markers that perform three and four functions in response to parasite Ags was observed in patients at a less severe disease stage. The frequency of CD8(+) T cells producing granzyme B and perforin and those expressing inhibitory receptors was higher in symptomatic patients than in asymptomatic patients. Taken together, these findings suggest that during the course of Chagas disease, CD8(+) T cells undergo a gradual loss of function characterized by impaired cytokine production, the presence of advanced differentiation, and increased inhibitory receptor coexpression.

[Nesting ecology of Crocodylus acutus (Reptilia: Crocodylidae) in Bahia Portete, La Guajira, Colombia]. / Ecología de la anidación de crocodylus acutus (ReptiliaL Crocodylidae) en Bahía Portete, La Guajira, Colombia.

C. acutus is cataloged in critical danger in Colombia. We studied its population at Bahia Portete, in order to survey the nesting activities and to obtain valuable information for a conservation management program. This study was undertaken with the participation of the Wayuü community using the Agreement Conservation Model, and took place during July 2007, August 2008, June and August 2009, and March to August 2010 and 2011. Sampling surveys were made by the use of transects along the coast, in order to find nests. For each nesting area found, we recorded the nests biometrics, eggs and hatchlings, fertility of the eggs and reproductive events. We explored a total of 55.12 km, and determined four nesting areas. Colonial nesting and the environmental variables were factors that affected the hatchling success. From the total of nests found, 37 % were oriented towards North, and 48 % were built next to Stenocereus griseus. Juyi island substrate was mostly constituted by sand (71.9 %, 71.44 %). The average number of eggs by nest was 28.42 ± 6.63, the largest egg diameter was 71.84 ± 3.54 mm, the average nest weight was 81.54 ± 9.99 g, and the hatchlings presented an average length of 25.47 ± 1.16 cm. For this specific site in Colombia, the reproductive period begins in March and ends in August. The few areas of nesting and the 95 % fertility suggest the performance of a conservation management program for this species at Bahía Portete.

Altering the motility of Trypanosoma cruzi with rabbit polyclonal anti-peptide antibodies reduces infection to susceptible mammalian cells.

Trypanosoma cruzi's trypomastigotes are highly active and their incessant motility seems to be important for mammalian host cell infection. The kinetoplastid membrane protein-11 (KMP-11) is a protein expressed in all parasite stages, which induces a cellular and humoral immune response in the infected host, and is hypothesized to participate in the parasite's motility. An N-terminal peptide from KMP-11, termed K1 or TcTLE, induced polyclonal antibodies that inhibit parasitic invasion of Vero cells. The goal of this study was to evaluate the motility and infectivity of T. cruzi when exposed to polyclonal anti-TcTLE antibodies. Rabbits were immunized with TcTLE peptide along with FIS peptide as an immunomodulator. ELISA assay results showed that post-immunization sera contained high titers of polyclonal anti-TcTLE antibodies, which were also reactive against the native KMP-11 protein and live parasites as detected by immunofluorescence and flow cytometry assays. Trypomastigotes of T. cruzi were incubated with pre- or post-immunization sera, and infectivity to human astrocytes was assessed by Giemsa staining/light microscope and flow cytometry using carboxyfluorescein diacetate succinimidyl ester (CFSE) labeled parasites. T. cruzi infection in astrocytes decreased approximately by 30% upon incubation with post-immunization sera compared with pre-immunization sera. Furthermore, trypomastigotes were recorded by video microscopy and the parasite's flagellar speed was calculated by tracking the flagella. Trypomastigotes exposed to post-immunization sera had qualitative alterations in motility and significantly slower flagella (45.5 µm/s), compared with those exposed to pre-immunization sera (69.2 µm/s). In summary, polyclonal anti-TcTLE serum significantly reduced the parasite's flagellar speed and cell infectivity. These findings support that KMP-11 could be important for parasite motility, and that by targeting its N-terminal peptide infectivity can be reduced.

Five-step authorship framework to improve transparency in disclosing contributors to industry-sponsored clinical trial publications.

Authorship guidelines have established criteria to guide author selection based on significance of contribution and helped to define associated responsibilities and accountabilities for the published findings. However, low awareness, variable interpretation, and inconsistent application of these guidelines can lead to confusion and a lack of transparency when recognizing those who merit authorship. This article describes a research project led by the Medical Publishing Insights and Practices (MPIP) Initiative to identify current challenges when determining authorship for industry-sponsored clinical trials and develop an improved approach to facilitate decision-making when recognizing authors from related publications. A total of 498 clinical investigators, journal editors, publication professionals and medical writers were surveyed to understand better how they would adjudicate challenging, real-world authorship case scenarios, determine the perceived frequency of each scenario and rate their confidence in the responses provided. Multiple rounds of discussions about these results with journal editors, clinical investigators and industry representatives led to the development of key recommendations intended to enhance transparency when determining authorship. These included forming a representative group to establish authorship criteria early in a trial, having all trial contributors agree to these criteria and documenting trial contributions to objectively determine who warrants an invitation to participate in the manuscript development process. The resulting Five-step Authorship Framework is designed to create a more standardized approach when determining authorship for clinical trial publications. Overall, these recommendations aim to facilitate more transparent authorship decisions and help readers better assess the credibility of results and perspectives of the authors for medical research more broadly. Please see related article: http://www.biomedcentral.com/1741-7015/12/214.

Collaborative measurement development as a tool in CBPR: measurement development and adaptation within the cultures of communities.

This paper describes the processes we engaged into develop a measurement protocol used to assess the outcomes in a community based suicide and alcohol abuse prevention project with two Alaska Native communities. While the literature on community-based participatory research (CBPR) is substantial regarding the importance of collaborations, few studies have reported on this collaboration in the process of developing measures to assess CBPR projects. We first tell a story of the processes around the standard issues of doing cross-cultural work on measurement development related to areas of equivalence. A second story is provided that highlights how community differences within the same cultural group can affect both the process and content of culturally relevant measurement selection, adaptation, and development.

Experiences of Workplace Violence in Graduate Nursing Clinical Education.

BACKGROUND: Exposure to workplace violence (WPV) is common in health care, and little is known about nurse practitioner (NP) students' experiences during graduate nursing clinical education. PURPOSE: This study described experiences of WPV among NP students during their clinical education. METHODS: We conducted a cross-sectional, quantitative survey of a random sample of NPs licensed in Texas examining graduate nursing education experiences. RESULTS: A total of 334 NPs responded, a 12% response rate. More than a quarter (27%) experienced WPV during their graduate nursing clinical experience. Preceptors were the most reported perpetrators (44%). Most NPs remained in their clinical site after their WPV experience (55%); a majority felt they had no alternative clinical placement option. CONCLUSIONS: Nurse practitioner students experience WPV, and there may be implications for educational persistence and their careers. Future work should attempt to reduce the frequency of student WPV experiences and examine implications for NP careers.

Operationalization of Clinical Placements in Graduate Nursing.

BACKGROUND: Nurse practitioner (NP) program accreditation standards require that programs secure clinical placements for all students. As NP programs increase enrollment to meet the demand for primary care providers, it is vital that they deploy a formalized clinical placement process that ensures all students have a clinical placement. PROBLEM: Although NP programs have consistently increased enrollment, the shortage of clinical sites and preceptors continues to be a barrier to admission. APPROACH: Described in this article is the operationalization of graduate nursing clinical placement at one large university with 7 NP tracks. OUTCOMES: A formalized clinical placement process ensures that all students receive an appropriate placement and graduate on time. Having a dedicated team of NP faculty members to provide clinical placements services for NP students is highly effective.

Pathophysiology and pharmacology course grades and prediction of success in a graduate nurse practitioner program.

BACKGROUND: During a program review, faculty identified that nurse practitioner (NP) students who received a C grade in Advanced Pathophysiology (Patho) and Advanced Pharmacology (Pharm) appeared to perform poorly in the later NP management courses and on other program outcomes. PURPOSE: The research aimed to determine whether grades in graduate Patho and Pharm courses could predict performance in NP management courses, program progression and completion, and certification pass rates. METHODOLOGY: This research included deidentified student data from 2016 to 2018 across seven NP specialty tracks ( n = 4,575). Nonparametric and parametric tests were used in the analysis. RESULTS: A significant correlation ( p < .001) existed between Patho and Pharm grades. Lower grades in these two courses were significantly related to each other and to lower management course grades. Logistic regression showed that graduate pathophysiology grades significantly predicted certification examination performance, with lower grades associated with lower certification examination performance. Graduate pharmacology grades, pathophysiology grades, composite management course grades, and admission grade point average (GPA) significantly predicted final cumulative GPA, with lower grades associated with lower performance for all variables. CONCLUSIONS: Results of this research support the hypothesis that grades of C in Patho or Pharm courses significantly predict C performance in the management NP courses and lower certification success rates. IMPLICATIONS: The project model can be used in future research. Study findings can be helpful to NP faculty when considering curriculum decisions.