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Purpose: This methodological study developed a scale that reflects the social support characteristics of nursing students who have experienced clinical practice and verified their reliability and validity. Methods: The 37 initial items of the scale were derived based on the results of a previous study that analyzed the concept of nursing students’ social support. The initial items were revised through content validity verification, and 29 preliminary items were finally selected. Data were collected from 220 students enrolled in a nursing department in Jeollabuk-do, who had clinical practice experience, and 205 surveys were used for the final analysis. The data collection period was from March 11 to April 26, 2022. An exploratory factor analysis was performed using maximum likelihood factor extraction and varimax rotation. Results: The social support scale for nursing students who have experienced clinical practice consisted of a total of three sub-factors and 17 items. The three sub-factors were ‘support from family and friends’ (eight items), ‘support from school, professors, and clinical instructors’ (seven items) and ‘support from department seniors’ (two items). The reliability of the developed scale was found to be high, with a Cronbach’s alpha of .93. As a result of verifying the criterion validity of the developed scale, the correlation between the criterion tool and the scale developed in this study was statistically significant. Conclusion: The significance of this study is that it developed a scale to measure social support for the first time among nursing students in Korea.

The estrogen receptor (ER/ESR1) is expressed in a majority of breast cancers and drugs that inhibit ER signaling are the cornerstone of breast cancer pharmacotherapy. Currently, aromatase inhibitors are the frontline endocrine interventions of choice although their durability in metastatic disease is limited by activating point mutations within the ligand-binding domain of ESR1 that permit ligand-independent activation of the receptor. It has been suggested that the most commonly occurring ESR1 mutations would likely compromise the clinical activity of selective estrogen receptor downregulators and selective estrogen receptor modulators (SERMs) when used as second-line therapies. It was unclear, however, how these mutations, which are likely coexpressed in cells with ERWT, may impact response to ER ligands in a clinically meaningful manner. To address this issue, we dissected the molecular mechanism(s) underlying ESR1-mutant pharmacology in models relevant to metastatic disease. These studies revealed that the response of ESR1 mutations to ligands was dictated primarily by the relative coexpression of ERWT in cells. Specifically, dysregulated pharmacology was only evident in cells in which the mutants were overexpressed relative to ligand-activated ERWT; a finding that highlights the role of allelism in determining ER-mutant pharmacology. Importantly, we demonstrated that the antagonist activity of the SERM, lasofoxifene, was not impacted by mutant status; a finding that has led to its clinical evaluation as a treatment for patients with advanced ER-positive breast cancer whose tumors harbor ESR1 mutations.

Supplemental Figure 1: Structures of clinically relevant compounds used in this study. Supplemental Figure 2: Cells expressing both the ERWT and ERmuts have similar pharmacological responses to antiestrogens when compared to cells only expressing ERWT. Supplemental Figure 3: Albeit differences in clonal variability, there is indistinguishable differences in T47D proliferation in response to antiestrogens. Supplemental Figure 4: Downregulation of ER in MCF7B cells is unaffected by the presence of the mutant receptor. Supplemental Figure 5: Validation of ER mutation status in engineered cell lines. Supplemental Figure 6: Protein levels of transfected constructs in SKBR3 cells are consistent with MCF7 expression level. Supplemental Figure 7: ER expression in MCF7I cells is titratable with doxycycline treatment. Supplemental Figure 8: Overexpression of ERY537S or ERD538G, not ERWT, in MCF7I cells changes ligand potency. Supplemental Figure 9: Western blot validation of relative ER expression levels in SKBR3. Supplemental Figure 10: Agonist activation of relative expression levels of ERWT and ERmuts in SKBR3 cells. Supplemental Figure 11: In-Cell Western validation of ER expression levels for siRNA knockdown and cDNA overexpression in MCF7i. Supplemental Figure 12: Agonist activation of relative expression levels of ERWT and ERmuts in MCF7i cells. Supplemental Figure 13: The altered potency of fulvestrant against the ERmuts on endogenous ER target genes is only evident when expressed at a level higher than the WT receptor.

Supplementary Figure from Comprehensive Analysis of Alternative Splicing in Gastric Cancer Identifies Epithelial–Mesenchymal Transition Subtypes Associated with Survival

Supplementary Data from Comprehensive Analysis of Alternative Splicing in Gastric Cancer Identifies Epithelial–Mesenchymal Transition Subtypes Associated with Survival

Supplementary Table from Comprehensive Analysis of Alternative Splicing in Gastric Cancer Identifies Epithelial–Mesenchymal Transition Subtypes Associated with Survival

Alternatively spliced RNA isoforms are a hallmark of tumors, but their nature, prevalence, and clinical implications in gastric cancer have not been comprehensively characterized. We systematically profiled the splicing landscape of 83 gastric tumors and matched normal mucosa, identifying and experimentally validating eight splicing events that can classify all gastric cancers into three subtypes: epithelial-splicing (EpiS), mesenchymal-splicing (MesS), and hybrid-splicing. These subtypes were associated with distinct molecular signatures and epithelial–mesenchymal transition markers. Subtype-specific splicing events were enriched in motifs for splicing factors RBM24 and ESRP1, which were upregulated in MesS and EpiS tumors, respectively. A simple classifier based only on RNA levels of RBM24 and ESRP1, which can be readily implemented in the clinic, was sufficient to distinguish gastric cancer subtypes and predict patient survival in multiple independent patient cohorts. Overall, this study provides insights into alternative splicing in gastric cancer and the potential clinical utility of splicing-based patient classification.Significance:This study presents a comprehensive analysis of alternative splicing in the context of patient classification, molecular mechanisms, and prognosis in gastric cancer.

Alternatively spliced RNA isoforms are a hallmark of tumors, but their nature, prevalence, and clinical implications in gastric cancer have not been comprehensively characterized. We systematically profiled the splicing landscape of 83 gastric tumors and matched normal mucosa, identifying and experimentally validating eight splicing events that can classify all gastric cancers into three subtypes: epithelial-splicing (EpiS), mesenchymal-splicing (MesS), and hybrid-splicing. These subtypes were associated with distinct molecular signatures and epithelial–mesenchymal transition markers. Subtype-specific splicing events were enriched in motifs for splicing factors RBM24 and ESRP1, which were upregulated in MesS and EpiS tumors, respectively. A simple classifier based only on RNA levels of RBM24 and ESRP1, which can be readily implemented in the clinic, was sufficient to distinguish gastric cancer subtypes and predict patient survival in multiple independent patient cohorts. Overall, this study provides insights into alternative splicing in gastric cancer and the potential clinical utility of splicing-based patient classification. Significance: This study presents a comprehensive analysis of alternative splicing in the context of patient classification, molecular mechanisms, and prognosis in gastric cancer.

Background Centers for Disease Control and Prevention (CDC) recommends 5 to 20 days of isolation for COVID-19 patients depending on symptom duration and severity regardless of genomic PCR results or vaccination history. However, in real clinical practice, more individualized approach is required. We thus developed clinical scoring system to predict viable viral shedding in a given patient by using various factors affecting viable viral shedding. Methods We prospectively enrolled adult patients with SARS-CoV-2 infection admitted to tertiary hospital and day care center between February 2020 and January 2022. The daily dense respiratory sampling (i.e. saliva, sputum, or nasopharyngeal swabs) during the hospital and day care center stay were obtained. Genomic RNA viral load and viral culture were performed for these samples. Clinical predictors of negative viral culture results were identified using survival analysis and multivariable analysis. Results A total of 612 samples from 121 patients of varying degrees of severity were obtained. Of these, 494 (81%) samples were saliva, 63 (10%) were nasopharyngeal swab, and the remaining 55 (9%) were sputum. Of these 612 specimens, 154 (25%) samples revealed positive viral culture results. Univariate and multivariable Cox's time varying proportional hazard model revealed that symptom onset day, viral copy number, disease severity, organ transplant recipient, gender, and vaccination status were independently associated with viral culture results. We thus developed the 5-factor model from -3 to 3 points: viral copy number (-3 to 3 points depending on copy number), disease severity (1 point to moderate to critical diseases), organ transplant recipient (2 points), gender (-1 points to male), and vaccination status (-2 points to fully vaccinated status). The predictive culture-negative rates were calculated through the symptom onset day and the score of the day the sample was collected. Table 1.Prediction scores for negative viral culture resultsTable 2.Predicted Culture-negative Rates Values in the table expressed as percentages. Conclusion Our clinical scoring system can provide objective probability of negative culture results in a given COVID-19 patient with genomic viral load, and appears to be useful to decide de-isolation policy depending on individualized factors associated with viable viral shedding beyond simple symptom-based isolation strategy by CDC. Disclosures All Authors: No reported disclosures.

Background Nontuberculous mycobacteria (NTM) species rarely cause vertebral osteomyelitis (VO). The clinical characteristics and radiological findings of NTM VO are not well described since the number of cases are limited. Here we present the clinical characteristics of culture positive NTM VO, compared to those of tuberculous (TB) VO. Methods We performed a retrospective, multicenter study at 5 hospitals. Patients with radiologic findings consistent with VO, and positive NTM or TB cultures on biopsy were included. Demographic characteristics and symptoms were collected, along with laboratory, radiologic, and pathologic findings. Results Nineteen patients with NTM VO and 105 patients with TB VO were included. M. intracellulare (52.6%) and M. abscessus (26.3%) were the most common NTM pathogens. The mean age of patients was significantly older in the NTM VO group (68.0 vs. 58.5 years, P = .01). Previous steroid or immunosuppressant use was more frequent in the NTM VO group (21.1% vs. 2.9%, P = .01; 15.8% vs. 2.9%, P = .046, respectively), as was a history of vertebral surgery (47.4% vs. 13.3%, P < .001). Symptoms and laboratory findings were similar between the 2 groups, although the proportion of patients with positive interferon-gamma release assays (IGRA) was significantly higher in the TB VO group (14.3% vs. 86.4%, P < .001). Radiologic findings were also similar, with lumbar involvement being the most common (57.9% and 44.8%, respectively). Chronic granulomatous inflammation was more frequently seen in TB VO patients (33.3% vs. 64.0%, P = .04), while inflammation alone was more common in NTM VO. Although there was no significant difference in survival after 1 year, survival at last follow-up (median, 24 months) was significantly lower in the NTM VO group (75.0% vs. 98.8%, P = .02). Conclusion NTM VO should be considered in older patients with a history of immunosuppression or previous vertebral surgery. The pathologic findings and radiologic findings of NTM VO were similar to that of TB VO. Granulomatous inflammation and positive IGRA tests were more common in TB than NTM VO. Disclosures All Authors: No reported disclosures.

Background The coronavirus disease 2019 (COVID-19) outbreak reached peak levels in South Korea as the Delta variant, dominant from late August until the end of 2021, was rapidly overtaken by the Omicron variant at the start of 2022. In studies conducted near the start of the pandemic, the occurrence of bacteremia in COVID-19 patients was relatively low. We aimed to determine if there was a change in the rate of bacteremia in COVID-19 patients with the progression of the pandemic. Methods We performed a retrospective study of patients who were diagnosed with COVID-19 at a referral hospital between September 2021 and March 2022. Blood culture results were recorded, along with demographic characteristics and clinical outcomes. Contamination was considered when a single blood culture was positive for coagulase-negative staphylococci (CoNS), Corynebacterium species, or Bacillus species. Clinically relevant bacteremia was defined as bacteremia due to clinically significant pathogens between 7 days before COVID-19 diagnosis to 14 days after diagnosis. Results Among the 360 patients included in the study, 46 cases from 43 (11.9%) patients were considered to be clinically relevant bacteremia. Enterococcus faecalis (17.4%) and CoNS (17.4%) were the most common pathogens, followed by Acinetobacter baumannii (15.2%), Staphylococcus aureus (10.7%) and Escherichia coli (10.7%). The median number of days from COVID-19 diagnosis to identification of bacteremia was 2 days. There was no significant difference in the rate of bacteremia between the Delta (September-December 2021) and Omicron variant eras (January-March 2022) (12.2% vs. 11.7%, P = .88). In the subgroup analysis of patients who received more than 2 days of intensive care, there was no statistical difference in the rate of bacteremia (14.5% [9/62] in the Delta variant era vs. 16.9% [14/83] in the Omicron variant era; P = .70). Mortality was significantly higher in patients with clinically relevant bacteremia (48.8% vs. 19.2%, P < .001). Conclusion Many COVID-19 patients had bacteremia in the Omicron variant era, especially in the intensive care unit. Clinicians should suspect bacterial co-infection when a COVID-19 patient is clinically aggravated. Disclosures All Authors: No reported disclosures.

Background We compared neutralizing antibodies in healthy healthcare workers against two different coronavirus 2019 disease (Covid-19) vaccines, a messenger RNA (mRNA)-based vaccine (BNT162b2) and a genetically modified organism (virus vector) vaccine (ChAdOx1), to obtain immunity. In our cohort, we tried to compare the level of neutralizing antibody production after a BNT162b2 vaccine booster and to analyze the level of neutralizing antibody titers for delta and omicron variant. Methods On November 23 to 25, 2021 and December 23 to 24, 2021, a total of 2,133 HCWs at Soonchunhyang University Bucheon Hospital had been vaccinated with BNT162b2 (Pfizer-BioNTech). Among the 115 HCWs who participated in the preceding study, all participants had no history of Covid-19 infection or suspected symptoms at the time of registration. We collected blood samples from participants four weeks after a third dose. All blood samples were analyzed using the commercial virus neutralization test kit (Genscript Biotech Corporation, Piscataway, NJ, USA). Results Titers were measured 2 months after the initial vaccination and before the booster of the BNT162b2 vaccine (i.e. 6 months after the initial vaccination), and 4 weeks after the booster. Neutralizing antibody level measured by percentage inhibition of surrogate virus neutralization test (sVNT) readings at negative control (unvaccinated) and 4 weeks after booster vaccination. BNT/BNT/BNT group was defined as a booster dose with BNT162b2 (Pfizer) after two doses of BNT162b2 vaccine (Pfizer) 3 weeks apart. ChA/ChA/BNT group was defined as a booster dose with BNT162b2 (Pfizer) after two doses of ChAdOx1 vaccine (AstraZeneca/Oxford) 12 weeks apart. Conclusion We found that the neutralizing antibody levels against omicron variant were higher among those who received a booster dose of the BNT162b2 vaccine. Disclosures All Authors: No reported disclosures.

Over half of BRAF mutant melanoma patients with response to targeted therapy recur within 15 months. Resistance is postulated to arise from certain clonal populations that enter a slow cycling persister state, evade treatment with relative dormancy, and repopulate the tumor when reactivated. Via longitudinal profiling we define expression states of clonal populations and track their evolutionary progression. BRAF mutant patient derived melanoma xenografts were treated with BRAF/MEK inhibitors through maximum tumor size reduction and re-growth. Spatial transcriptomics (ST) was performed at 5 timepoints across 94 days of treatment, tracking clonal populations with intact tissue structure. Deep learning on H&E-stained images automatically detected histological features that co-localize with expression levels. A novel computational pipeline performed clustering, differential expression and pathway enrichment analysis, copy number variation detection, and pseudotemporal ordering, allowing spatial recreation of clonal phylogenies and cell fate trajectory inference during the treatment course. Three clonal categories were defined; Sensitive: predominant in treatment-naïve samples, Persister: remaining clusters in maximally shrunken specimens, Resistant: re-emergent, fast-proliferating. Analysis of clonal composition identified GAPDH and CCND1 as highly expressed in sensitive clones, suppressed in persisters, and reactivated in resistant clones; TYRP1, DCT, MITF, and NGFR showed an opposite pattern. Persister clones evolved from oxidative phosphorylation toward glycolysis and MAPK regulation. Spatial analysis showed increased glycolysis:oxidative phosphorylation ratio in centrally located sensitive clusters, with more balanced peripheral expression. Resistant clones showed upregulated DUSP6 expression and enrichment in Orexin signaling and MAPK regulation. Imaging- and expression-based clones were highly concordant. Spatial profiling during melanoma treatment identified transient persister and emergent resistant clones, defining expression profiles and phenotypic features. Altered ratio of oxidative to glycolytic metabolic pathways was a hallmark of clonal evolution in both space and time. Specific MAPK pathway re-entry points were identified as candidate resistance mechanisms, suggesting potential therapeutic targets. Deep learning features derived from histological images showed good correlation with ST profiles, providing a promising method for clonal tracking via images alone. This longitudinal experiment mimics a tumor’s clinical course, inducing persistence and resistance via treatment that parallels that seen by patients. Combining ST and imaging techniques, we provide insight into clonal dynamics with novel spatiotemporal resolution, defining an evolutionary roadmap to acquired treatment resistance. Citation Format: Jill Carol Rubinstein, Sergii Domanskyi, Todd B. Sheridan, Brian J. Sanderson, SungHee Park, Jessica Kaster, Haiyin Li, Olga Anczukow, Meenhard Herlyn, Jeffrey H. Chuang. Spatiotemporal profiling defines persister and resistance signatures in targeted treatment of melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1164.

Eosinophilic meningitis is defined as the presence of more than 10 eosinophils per μl in the cerebrospinal fluid (CSF), or eosinophils accounting for more than 10% of CSF leukocytes in patients with acute meningitis. Parasites are the most common cause of eosinophilic meningitis worldwide, but there is limited research on patients in Korea. Patients diagnosed with eosinophilic meningitis between January 2004 and June 2018 at a tertiary hospital in Seoul, Korea were retrospectively reviewed. The etiology and clinical characteristics of each patient were identified. Of the 22 patients included in the study, 11 (50%) had parasitic causes, of whom 8 (36%) were diagnosed as neurocysticercosis and 3 (14%) as Toxocara meningitis. Four (18%) patients were diagnosed with fungal meningitis, and underlying immunodeficiency was found in 2 of these patients. The etiology of another 4 (18%) patients was suspected to be tuberculosis, which is endemic in Korea. Viral and bacterial meningitis were relatively rare causes of eosinophilic meningitis, accounting for 2 (9%) and 1 (5%) patients, respectively. One patient with neurocysticercosis and 1 patient with fungal meningitis died, and 5 (23%) had neurologic sequelae. Parasite infections, especially neurocysticercosis and toxocariasis, were the most common cause of eosinophilic meningitis in Korean patients. Fungal meningitis, while relatively rare, is often aggressive and must be considered when searching for the cause of eosinophilic meningitis.

Rebaudioside A is a promising natural alternative sweetener but they produce increased bitterness, astringency, and unpleasant aftertastes. Glycosylation and blending with different sweeteners are known to improve the sensory characteristics of rebaudioside A. The present study was conducted to identify the relative sweetness and sensory profile of glycosyl rebaudioside A (g-reb A). The relative sweetness of g-reb A compared to 5% sucrose was determined using the two-alternative forced choice method. The sensory profiles of g-reb A and its mixtures with allulose and maltitol (1:1 ratio) were compared to those of rebaudioside A, rebaudioside D, rebaudioside M, sucralose, allulose, maltitol, and sucrose using descriptive analysis conducted by eight trained panelists. The relative sweetness of g-reb A was 155, which was lower than that of rebaudioside A. In addition, the bitter taste and aftertaste, astringency, and sweet onset of g-reb A were decreased compared to those of rebaudioside A.

The estrogen receptor (ER/ESR1) is expressed in a majority of breast cancers and drugs that inhibit ER signaling are the cornerstone of breast cancer pharmacotherapy. Currently, aromatase inhibitors are the frontline endocrine interventions of choice although their durability in metastatic disease is limited by activating point mutations within the ligand-binding domain of ESR1 that permit ligand-independent activation of the receptor. It has been suggested that the most commonly occurring ESR1 mutations would likely compromise the clinical activity of selective estrogen receptor downregulators and selective estrogen receptor modulators (SERMs) when used as second-line therapies. It was unclear, however, how these mutations, which are likely coexpressed in cells with ERWT, may impact response to ER ligands in a clinically meaningful manner. To address this issue, we dissected the molecular mechanism(s) underlying ESR1-mutant pharmacology in models relevant to metastatic disease. These studies revealed that the response of ESR1 mutations to ligands was dictated primarily by the relative coexpression of ERWT in cells. Specifically, dysregulated pharmacology was only evident in cells in which the mutants were overexpressed relative to ligand-activated ERWT; a finding that highlights the role of allelism in determining ER-mutant pharmacology. Importantly, we demonstrated that the antagonist activity of the SERM, lasofoxifene, was not impacted by mutant status; a finding that has led to its clinical evaluation as a treatment for patients with advanced ER-positive breast cancer whose tumors harbor ESR1 mutations.

Purpose: The purpose of this study was to explore the experiences of perception of nursing students' rights participating in clinical practice. Methods: This descriptive study carried out purposeful sampling. The participants were 17 nursing students, who had experience of participating in clinical practice for more than 12 weeks. Data were collected through focus group interviews. Twelve subjects were in their third year and five in their fourth year. All were practicing in secondary general and tertiary university hospitals. The data were analyzed using classical content analysis method. Results: The researchers extracted 23 codes representing the nursing students' rights of clinical practice, which were grouped into 4 categories and 11 subcategories. The 4 categories were 'deep disappointment as an alienated person in a clinical field', 'clinical practice experience that cannot be given up despite difficulties', 'need for a practice environment that takes care of nursing students', and 'hope for support, advocacy and respect'. Conclusion: Nursing students cannot claim rights at this time, but expressed the desire to build a support system so that these parts can be improved in the future. Therefore, nursing education institutions and clinical fields should maintain diverse efforts through reciprocal relationships.

Summary: Misregulation of alternative splicing is a hallmark of human tumors, yet to what extent and how it contributes to malignancy are only beginning to be unraveled. Here, we define which members of the splicing factor SR and SR-like families contribute to breast cancer and uncover differences and redundancies in their targets and biological functions. We identify splicing factors frequently altered in human breast tumors and assay their oncogenic functions using breast organoid models. We demonstrate that not all splicing factors affect mammary tumorigenesis in MCF-10A cells. Specifically, the upregulation of SRSF4, SRSF6, or TRA2β disrupts acinar morphogenesis and promotes cell proliferation and invasion in MCF-10A cells. By characterizing the targets of these oncogenic splicing factors, we identify shared spliced isoforms associated with well-established cancer hallmarks. Finally, we demonstrate that TRA2β is regulated by the MYC oncogene, plays a role in metastasis maintenance in vivo, and its levels correlate with breast cancer patient survival. : Park et al. demonstrate that >50% of human breast tumors exhibit an alteration in one of the splicing factors from the SR protein family. Using in vitro and in vivo breast cancer models, they identify three splicing factors that promote cell proliferation and invasion by regulating isoforms associated with cancer hallmarks. Keywords: alternative RNA splicing, breast cancer, splicing factor, SR protein, metastasis, MYC, TRA2-beta, triple negative breast cancer

MYC is dysregulated in >50% of cancers, but direct targeting of MYC has been clinically unsuccessful. Targeting downstream MYC effector pathways represents an attractive alternative. MYC regulates alternative mRNA splicing, a hallmark of cancer, but the mechanistic links between MYC and the splicing machinery remain underexplored. Here, we identify a network of splicing factors (SFs) co-expressed as SF-modules in MYC-active breast tumors. Of these, one is a pan-cancer SF-module, correlating with MYC-activity across 33 tumor types. In mammary cell models, MYC activation leads to co-upregulation of pan-cancer module SFs and to changes in >4,000 splicing events. In breast cancer organoids, co-overexpression of the pan-cancer SF-module is sufficient to induce splicing events that are also MYC-regulated in patient tumors and to increase organoid size and invasiveness, while its knockdown decreases organoid size. Finally, we uncover a pan-cancer splicing signature of MYC activity which correlates with survival in multiple tumor types. Our findings provide insight into the mechanisms and function of MYC-regulated splicing and for the development of therapeutics for MYC-driven tumors.

Although general translation declines during fasting, maintaining the translation of a subset or proteins is necessary for metabolic homeostasis and cell viability. Using unbiased proteome analysis of hepatic cells during starvation, we identified a novel pathway in which Esrra-mediated transcription of Rplp1-dependent translation of lysosomal proteins declined during early starvation and recovered after prolonged starvation to restore autophagy-lysosome function. Interestingly, hepatic Esrra-Rplp1-dependent translation rate of lysosomal proteins also was impaired in patients and mice with non-alcoholic steatohepatitis (NASH), and translational response to starvation was dysregulated in mice with NASH. Remarkably, activation of Esrra pharmacologically, genetically, or by alternate day fasting restored protein translation, increased expression of lysosomal proteins, induced autophagy, and reduced lipotoxicity, inflammation, and fibrosis in cell culture andin vivomodels of NASH. Thus, hepatic Esrra is essential for ribosome-dependent translation of lysosomal proteins during starvation, and prevention of lipotoxicity and progression in NASH.Lay summaryFasting for weight loss improves non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH); however, the mechanism is not well understood. Our study shows that a nuclear protein, estrogen related receptor alpha (Esrra), increases ribosome-mediated translation of autophagy and lysosome proteins during chronic starvation to maintain essential metabolic pathways for cell survival. Surprisingly, this translational pathway is impaired during NASH with reduced lysosome-autophagy activity accompanied by increased inflammation and fibrosis gene expression in the liver. Pharmacologic, genetic and dietary activation of Esrra decreases lipid-mediated toxicity in liver cells as well as inflammation and fibrosis in livers from mice with NASH. These findings suggest that the Esrra-ribosome-lysosome pathway is important for liver response to fasting and NASH and thus may be a good therapeutic target for the treatment of NASH.

Background Data on the clinical and virological characteristics of the Delta variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are limited. This prospective cohort study compared the characteristics of the Delta variant to other variants. Methods Adult patients with mild coronavirus disease 2019 (COVID-19) who agreed to daily saliva sampling at a community isolation facility in South Korea between July and August 2021 were enrolled. Scores of 28 COVID-19-related symptoms were recorded daily. The genomic RNA and subgenomic RNA from saliva samples were measured by real-time reverse-transcription polymerase chain reaction (PCR). Cell cultures were performed on saliva samples with positive genomic RNA results. Results A total of 141 patients (Delta group, n = 108 [77%]; non-Delta group, n = 33 [23%]) were enrolled. Myalgia was more common in the Delta group than in the non-Delta group (52% vs 27%, P = .03). Total symptom scores were significantly higher in the Delta group between days 3 and 10 after symptom onset. Initial genomic RNA titers were similar between the 2 groups; however, during the late course of disease, genomic RNA titers were higher in the Delta group. Negative conversion of subgenomic RNA was slower in the Delta group (median 9 vs 5 days; P Conclusions COVID-19 patients infected with the Delta variant exhibited prolonged viable viral shedding with more severe symptoms than those infected with non-Delta variants.

MYC is dysregulated in50% of cancers, but direct targeting of MYC has been clinically unsuccessful. Targeting downstream MYC effector pathways represents an attractive alternative. MYC regulates alternative mRNA splicing, but the mechanistic links between MYC and the splicing machinery in cancer remain underexplored. Here, we identify a network of co-expressed splicing factors (SF-modules) in MYC-active breast tumors. Of these, one is a pan-cancer SF-module correlating with MYC activity across 33 tumor types. In mammary cell models, MYC activation leads to co-upregulation of pan-cancer module SFs and to changes in4,000 splicing events. In breast cancer organoids, co-overexpression of the pan-cancer SF-module induces MYC-regulated splicing events and increases organoid size and invasiveness, while knockdown decreases organoid size. Finally, we uncover a MYC-activity pan-cancer splicing signature correlating with survival across tumor types. Our findings provide insight into the mechanisms of MYC-regulated splicing and for the development of therapeutics for MYC-driven tumors.

The oncogenic transcription factor MYC is one of the most frequently altered genes in cancer and is highly dysregulated in aggressive tumor types, including triple negative breast cancer (TNBC). Although direct targeting of MYC or its regulators in human tumors has proved difficult, the undisputed clinical relevance of this transcription factor warrants continued study for therapeutic targeting of MYC-driven processes in tumors. Recently, we and others have shown the tumorigenic capacity of MYC is critically dependent on the RNA splicing machinery and on several splicing factor RNA-binding proteins (RBPs). Dysregulation of alternative mRNA splicing, a key step in gene expression regulation, is a hallmark of cancer, and can lead to the expression of isoforms driving tumor initiation, progression or drug resistance. Understanding which component(s) of the RNA machinery are regulated by MYC, and the functions of MYC-induced spliced isoforms, may reveal new targets that circumvent prior failures of directly targeting MYC. Here, we examine MYC-induced changes in gene expression and alternative splicing through RNA-sequencing of breast cell lines and tumors. Using a mammary epithelial cell line with an inducible MYC system, we identified over 4,000 MYC-induced alternative splicing events as well as 378 MYC-induced RBPs. Co-expression network analysis of these RBPs in TCGA breast tumor RNA-seq data uncovered 8 RBP modules, four of which are highly correlated with MYC activity and highly expressed in TNBC. One of these modules is preserved across almost all other TCGA tumor types and remains highly correlated with MYC activity, suggesting a pan-cancer role of this co-expression module. We are now defining the functional role of these RBP modules in MYC-driven tumorigenesis in breast cancer models, as well as identifying their downstream oncogenic splicing targets. Our research project will define the role and regulation of alternative splicing in breast cancer, and more specifically in aggressive tumor types such as TNBC. Our findings will provide mechanistic insight into the role of MYC-driven splicing in tumor maintenance and will uncover novel therapeutic targets for MYC-active tumors. Citation Format: Laura Urbanski, Brittany Angarola, Mattia Brugiolo, Marina Yurieva, Sunghee Park, Olga Anczukow. Myc regulates alternative splicing through a network of RNA binding proteins in breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PD15-07.

Nursing students perform clinical training in a variety of clinical practice fields, so their rights are important. Efforts are needed to accurately identify and measure the awareness of nursing students’ rights. This study aimed to develop and evaluate the psychometric properties of nursing students’ rights awareness scale. The study procedure was carried out in four stages: the concept analysis, item development, scale development, and scale evaluation stage. First, in the concept analysis stage, the attributes of the concept were derived. Second, in the item development stage, preliminary items were derived, and the content validity was verified. Third, in the scale development stage, a preliminary and main survey were conducted, and item reduction was implemented. Fourth, in the scale evaluation stage, construct validity and reliability were verified. The collected data were analyzed using the SPSS 24.0, and item analysis was conducted using skewness, kurtosis, and item-total correlation scores. The construct validity was assessed by exploratory factor analysis, convergent validity, and divergent validity. The reliability was analyzed using Cronbach’s alpha coefficient and composite reliability. The final version of the scale was tested on 240 nursing students from three cities in South Korea. From the construct validity, three factors and 14 items were selected. The factors included “the rights to be protected, cared for, supported, and respected” (eight items), “the rights to be recognized as a member of a nursing team” (three items), and “the rights to learn” (three questions). The reliability of the scale was also verified. Through this study, the scale was developed to identify the rights of nursing students in clinical practice. The implication of this study is that it has laid the foundation for measuring the rights of nursing students applicable in clinical settings for the first time. The limitation of this study is that data were collected in some regions. Therefore, it is necessary to verify the validity and reliability of the scale in various cultures.

Summary: Most cancer cells exhibit metabolic flexibility, enabling them to withstand fluctuations in intratumoral concentrations of glucose (and other nutrients) and changes in oxygen availability. While these adaptive responses make it difficult to achieve clinically useful anti-tumor responses when targeting a single metabolic pathway, they can also serve as targetable metabolic vulnerabilities that can be therapeutically exploited. Previously, we demonstrated that inhibition of estrogen-related receptor alpha (ERRα) significantly disrupts mitochondrial metabolism and that this results in substantial antitumor activity in animal models of breast cancer. Here we show that ERRα inhibition interferes with pyruvate entry into mitochondria by inhibiting the expression of mitochondrial pyruvate carrier 1 (MPC1). This results in a dramatic increase in the reliance of cells on glutamine oxidation and the pentose phosphate pathway to maintain nicotinamide adenine dinucleotide phosphate (NADPH) homeostasis. In this manner, ERRα inhibition increases the efficacy of glutaminase and glucose-6-phosphate dehydrogenase inhibitors, a finding that has clinical significance. : Park et al. demonstrate that inhibition of mitochondrial metabolism can be accomplished using small molecule inhibitors of ERRα. Inhibiting the activity of this receptor decreases the expression of MPC1, interferes with pyruvate entry into the mitochondria, and increases cellular reliance on glutamine oxidation and the pentose phosphate pathway (PPP) to maintain NADPH homeostasis. Keywords: nuclear receptor, ERR⍺, mitochondrial metabolism, NADPH, glutaminase, pentose phosphate pathway, MPC1, oxidative stress, breast cancer, pyruvate carrier