RESUMO
Human cytomegalovirus (HCMV) replication requires host metabolism. Infection alters the activity in multiple metabolic pathways, including increasing fatty acid elongation and lipid synthesis. The virus-host interactions regulating the metabolic changes associated with replication are essential for infection. While multiple host factors, including kinases and transcription factors, important for metabolic changes that occur following HCMV infection have been identified, little is known about the viral factors required to alter metabolism. In this study, we tested the hypothesis that pUL37x1 is important for the metabolic remodeling that is necessary for HCMV replication using a combination of metabolomics, lipidomics, and metabolic tracers to measure fatty acid elongation. We observed that fibroblast cells infected with wild-type (WT) HCMV had levels of metabolites similar to those in cells infected with a mutant virus lacking the UL37x1 gene, subUL37x1. However, we found that relative to WT-infected cells, subUL37x1-infected cells had reduced levels of two host proteins that were previously demonstrated to be important for lipid metabolism during HCMV infection: fatty acid elongase 7 (ELOVL7) and the endoplasmic reticulum (ER) stress-related kinase PERK. Moreover, we observed that HCMV infection results in an increase in phospholipids with very-long-chain fatty acid tails (PL-VLCFAs) that contain 26 or more carbons in one of their two tails. The levels of many PL-VLCFAs were lower in subUL37x1-infected cells than in WT-infected cells. Overall, we conclude that although pUL37x1 is not necessary for network-wide metabolic changes associated with HCMV infection, it is important for the remodeling of a subset of metabolic changes that occur during infection.IMPORTANCE Human cytomegalovirus (HCMV) is a common pathogen that asymptomatically infects most people and establishes a lifelong infection. However, HCMV can cause end-organ disease that results in death in the immunosuppressed and is a leading cause of birth defects. HCMV infection depends on host metabolism, including lipid metabolism. However, the viral mechanisms for remodeling of metabolism are poorly understood. In this study, we demonstrate that the viral UL37x1 protein (pUL37x1) is important for infection-associated increases in lipid metabolism, including fatty acid elongation to produce very-long-chain fatty acids (VLCFAs). Furthermore, we found that HCMV infection results in a significant increase in phospholipids, particularly those with VLCFA tails (PL-VLCFAs). We found that pUL37x1 was important for the high levels of fatty acid elongation and PL-VLCFA accumulation that occur in HCMV-infected cells. Our findings identify a viral protein that is important for changes in lipid metabolism that occur following HCMV infection.
Assuntos
Infecções por Citomegalovirus/metabolismo , Citomegalovirus/fisiologia , Proteínas Imediatamente Precoces/metabolismo , Metabolismo dos Lipídeos , Linhagem Celular , Citomegalovirus/genética , Citomegalovirus/metabolismo , Infecções por Citomegalovirus/virologia , Elongases de Ácidos Graxos/metabolismo , Ácidos Graxos/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Proteínas Imediatamente Precoces/genética , Lipidômica , Metaboloma , Mutação , Fosfolipídeos/química , Fosfolipídeos/metabolismo , Replicação Viral , eIF-2 Quinase/metabolismoRESUMO
OBJECTIVE: To evaluate the impact of the recent changes to the urology residency application process on the criteria utilized by residency program directors (PDs) for interview invitations and their perspectives concerning these changes. METHODS: One hundred thirty-seven urology residency PDs were invited to participate in an anonymous survey to explore interview selection criteria and the impact of the increase in preference signals (PS) per applicant. RESULTS: Fifty-eight PDs (42.8%) completed the survey. The highest-ranked criteria were letters of recommendation (LoR) and successful sub-internship (sub-I) at the PD's institution, without statistically significant differences between these 2. Gender, ethnicity, and medical school prestige were the lowest rated criteria, without significant differences between these 3. Compared to before the increase in the number of PS per applicant, 80.7% of PDs reported that not receiving a PS from an applicant this cycle would more negatively impact the chances of offering an interview to that applicant. Moreover, 12.2% stated they would not interview any applicants who did not send a PS. Finally, 62.1% of PDs believed recent changes worsened the process. CONCLUSION: Recent changes impacted PDs applicant evaluation, with the highest ranked criteria being LoRs and sub-I. Paradoxically, the increase in the number of PS per applicant has increased their importance as applicants are much less likely to receive interview offers from programs they have not signaled. Lastly, most PDs believe changes have worsened the evaluation process.
Assuntos
Internato e Residência , Seleção de Pessoal , Urologia , Urologia/educação , Humanos , Masculino , Feminino , Seleção de Pessoal/normas , Seleção de Pessoal/métodos , Critérios de Admissão Escolar , Inquéritos e Questionários , Diretores Médicos , Entrevistas como AssuntoRESUMO
OBJECTIVE: To assess the rationale behind the choice of programs for preference signaling (PS) and subinternships by urology applicants in the 2023 cycle. METHODS: We emailed an anonymous, multiple-choice survey to the 403 prospective candidates who applied to our institution for the 2023 Urology Residency Match. RESULTS: 121 applicants (30.0%) responded to the survey. 81.8% were in favor of the continuation of PS, with 4.1% against it. When choosing where to PS or subinternship, geographic location and perceived quality of clinical training were the highest-ranked criteria. Racial/ethnic diversity of the residents influenced PS or subinternship choice for 77.8% of Black, 72.7% of Asian, 57.1% of Latinos, and 46.5% of White respondents (P < .05 for Black and Asian vs White). Institutional statements on diversity influenced PS or subinternship choice for 88.9% of Black, 55% of Asian, 64.3% of Latino, and 25.4% of White respondents (P < .05 for Black, Asian and Latino vs White). Females had an increased likelihood of PS or choosing subinternship programs with gender diversity of residents (81.6% vs 33.8, P = .002). A program with PS was associated with a 2.74 increase in likelihood of obtaining an interview compared to programs that were not PS. Of 107 matched applicants, 47.5% matched at a program where they completed a subinternship, and 25.7% matched at a signaled program. CONCLUSION: PS plays a major role in the match process. When choosing where to PS, applicants prioritize geographic location and perceived quality of training. Diversity of residents plays a major role in recruiting diverse applicants.