RESUMEN
Investigating therapeutic "outliers" that show exceptional responses to anti-cancer treatment can uncover biomarkers of drug sensitivity. We performed preclinical trials investigating primary murine acute myeloid leukemias (AMLs) generated by retroviral insertional mutagenesis in KrasG12D "knockin" mice with the MEK inhibitor PD0325901 (PD901). One outlier AML responded and exhibited intrinsic drug resistance at relapse. Loss of wild-type (WT) Kras enhanced the fitness of the dominant clone and rendered it sensitive to MEK inhibition. Similarly, human colorectal cancer cell lines with increased KRAS mutant allele frequency were more sensitive to MAP kinase inhibition, and CRISPR-Cas9-mediated replacement of WT KRAS with a mutant allele sensitized heterozygous mutant HCT116 cells to treatment. In a prospectively characterized cohort of patients with advanced cancer, 642 of 1,168 (55%) with KRAS mutations exhibited allelic imbalance. These studies demonstrate that serial genetic changes at the Kras/KRAS locus are frequent in cancer and modulate competitive fitness and MEK dependency.
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Antineoplásicos/uso terapéutico , Benzamidas/uso terapéutico , Neoplasias Colorrectales/genética , Difenilamina/análogos & derivados , Sistema de Señalización de MAP Quinasas , Proteínas Proto-Oncogénicas p21(ras)/genética , Animales , Antineoplásicos/farmacología , Benzamidas/farmacología , Línea Celular Tumoral , Evolución Clonal , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/patología , Difenilamina/farmacología , Difenilamina/uso terapéutico , Resistencia a Antineoplásicos , Humanos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Mutación , RetroviridaeRESUMEN
The COVID-19 pandemic has disproportionately affected racial and ethnic minority groups in the United States. Whereas racial and ethnic disparities in severe COVID-19-associated outcomes, including mortality, have been documented (1-3), less is known about population-based disparities in infection with SARS-CoV-2, the virus that causes COVID-19. In addition, although persons aged <30 years account for approximately one third of reported infections,§ there is limited information on racial and ethnic disparities in infection among young persons over time and by sex and age. Based on 689,672 U.S. COVID-19 cases reported to CDC's case-based surveillance system by jurisdictional health departments, racial and ethnic disparities in COVID-19 incidence among persons aged <25 years in 16 U.S. jurisdictions¶ were described by age group and sex and across three periods during January 1-December 31, 2020. During January-April, COVID-19 incidence was substantially higher among most racial and ethnic minority groups compared with that among non-Hispanic White (White) persons (rate ratio [RR] range = 1.09-4.62). During May-August, the RR increased from 2.49 to 4.57 among non-Hispanic Native Hawaiian and Pacific Islander (NH/PI) persons but decreased among other racial and ethnic minority groups (RR range = 0.52-2.82). Decreases in disparities were observed during September-December (RR range = 0.37-1.69); these decreases were largely because of a greater increase in incidence among White persons, rather than a decline in incidence among racial and ethnic minority groups. NH/PI, non-Hispanic American Indian or Alaska Native (AI/AN), and Hispanic or Latino (Hispanic) persons experienced the largest persistent disparities over the entire period. Ensuring equitable and timely access to preventive measures, including testing, safe work and education settings, and vaccination when eligible is important to address racial/ethnic disparities.
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COVID-19/etnología , Etnicidad/estadística & datos numéricos , Disparidades en el Estado de Salud , Grupos Minoritarios/estadística & datos numéricos , Grupos Raciales/estadística & datos numéricos , Adolescente , Distribución por Edad , Niño , Preescolar , Femenino , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Distribución por Sexo , Factores de Tiempo , Estados Unidos/epidemiología , Adulto JovenRESUMEN
There is accumulating evidence that functional alteration(s) of the bone marrow (BM) microenvironment contribute to the development of some myeloid disorders, such as myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). In addition to a cell-intrinsic role of WNT activation in leukemia stem cells, WNT activation in the BM niche is also thought to contribute to the pathogenesis of MDS and AML. We previously showed that the Apc-haploinsufficient mice (Apcdel/+ ) model MDS induced by an aberrant BM microenvironment. We sought to determine whether Apc, a multifunctional protein and key negative regulator of the canonical ß-catenin (Ctnnb1)/WNT-signaling pathway, mediates this disease through modulating WNT signaling, and whether inhibition of WNT signaling prevents the development of MDS in Apcdel/+ mice. Here, we demonstrate that loss of 1 copy of Ctnnb1 is sufficient to prevent the development of MDS in Apcdel/+ mice and that altered canonical WNT signaling in the microenvironment is responsible for the disease. Furthermore, the US Food and Drug Administration (FDA)-approved drug pyrvinium delays and/or inhibits disease in Apcdel/+ mice, even when it is administered after the presentation of anemia. Other groups have observed increased nuclear CTNNB1 in stromal cells from a high frequency of MDS/AML patients, a finding that together with our results highlights a potential new strategy for treating some myeloid disorders.
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Genes APC , Síndromes Mielodisplásicos/genética , Síndromes Mielodisplásicos/prevención & control , Nicho de Células Madre/genética , Vía de Señalización Wnt , Animales , Modelos Animales de Enfermedad , Haploinsuficiencia , Humanos , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/patología , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Síndromes Mielodisplásicos/patología , Compuestos de Pirvinio/farmacología , Vía de Señalización Wnt/genética , beta Catenina/genéticaRESUMEN
BRCA1 is critical for maintenance of genomic stability and interacts directly with several proteins that regulate hematopoietic stem cell function and are part of the Fanconi anemia (FA) double-strand break DNA repair pathway. The effects of complete BRCA1 deficiency on bone marrow (BM) function are unknown. To test the hypothesis that Brca1 is essential in hematopoiesis, we developed a conditional mouse model with Mx1-Cre-mediated Brca1 deletion. Mice lacking Brca1 in the BM have baseline cytopenias and develop spontaneous bone marrow failure or diverse hematologic malignancies by 6 months of age. Brca1(-/-) BM cells have a reduced capacity to form hematopoietic colonies in vitro and to reconstitute hematopoiesis in irradiated recipients, consistent with a hematopoietic progenitor functional defect. Brca1(-/-) BM cells also show FA-like hypersensitivity to the DNA crosslinking agent mitomycin C, and karyotypes feature genomic instability. Taken together, our results show that loss of Brca1 in murine BM causes hematopoietic defects similar to those seen in people with FA, which provides strong evidence that Brca1 is critical for normal hematopoiesis and that Brca1 is a bona fide FA-like gene.
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Proteína BRCA1/deficiencia , Proteína BRCA1/genética , Neoplasias Hematológicas/genética , Hemoglobinuria Paroxística/genética , Anemia Aplásica , Animales , Médula Ósea/patología , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/patología , Enfermedades de la Médula Ósea , Trastornos de Fallo de la Médula Ósea , Modelos Animales de Enfermedad , Inestabilidad Genómica , Neoplasias Hematológicas/diagnóstico , Hematopoyesis/genética , Hemoglobinuria Paroxística/diagnóstico , Humanos , Infiltración Leucémica , Recuento de Leucocitos , Ratones , Ratones Noqueados , Pancitopenia/genética , Pancitopenia/patología , FenotipoRESUMEN
Deregulated activation of ß-catenin in cancer has been correlated with genomic instability. During thymocyte development, ß-catenin activates transcription in partnership with T-cell-specific transcription factor 1 (Tcf-1). We previously reported that targeted activation of ß-catenin in thymocytes (CAT mice) induces lymphomas that depend on recombination activating gene (RAG) and myelocytomatosis oncogene (Myc) activities. Here we show that these lymphomas have recurring Tcra/Myc translocations that resulted from illegitimate RAG recombination events and resembled oncogenic translocations previously described in human T-ALL. We therefore used the CAT animal model to obtain mechanistic insights into the transformation process. ChIP-seq analysis uncovered a link between Tcf-1 and RAG2 showing that the two proteins shared binding sites marked by trimethylated histone-3 lysine-4 (H3K4me3) throughout the genome, including near the translocation sites. Pretransformed CAT thymocytes had increased DNA damage at the translocating loci and showed altered repair of RAG-induced DNA double strand breaks. These cells were able to survive despite DNA damage because activated ß-catenin promoted an antiapoptosis gene expression profile. Thus, activated ß-catenin promotes genomic instability that leads to T-cell lymphomas as a consequence of altered double strand break repair and increased survival of thymocytes with damaged DNA.
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Inestabilidad Genómica , Activación de Linfocitos , Linfoma/genética , Linfocitos T/citología , beta Catenina/metabolismo , Animales , Apoptosis , Secuencia de Bases , Supervivencia Celular , Roturas del ADN de Doble Cadena , Metilación de ADN , Reparación del ADN , Modelos Animales de Enfermedad , Genes RAG-1/genética , Factor Nuclear 1-alfa del Hepatocito , Histonas/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Recombinación Genética , Factor 1 de Transcripción de Linfocitos T/metabolismo , Timocitos/citología , Translocación Genética , beta Catenina/genéticaRESUMEN
An interstitial deletion of chromosome 5, del(5q), is the most common structural abnormality in primary myelodysplastic syndromes (MDS) and therapy-related myeloid neoplasms (t-MNs) after cytotoxic therapy. Loss of TP53 activity, through mutation or deletion, is highly associated with t-MNs with a del(5q). We previously demonstrated that haploinsufficiency of Egr1 and Apc, 2 genes lost in the 5q deletion, are key players in the progression of MDS with a del(5q). Using genetically engineered mice, we now show that reduction or loss of Tp53 expression, in combination with Egr1 haploinsufficiency, increased the rate of development of hematologic neoplasms and influenced the disease spectrum, but did not lead to overt myeloid leukemia, suggesting that altered function of additional gene(s) on 5q are likely required for myeloid leukemia development. Next, we demonstrated that cell intrinsic loss of Tp53 in hematopoietic stem and progenitor cells haploinsufficient for both Egr1 and Apc led to the development of acute myeloid leukemia (AML) in 17% of mice. The long latency (234-299 days) and clonal chromosomal abnormalities in the AMLs suggest that additional genetic changes may be required for full transformation. Thus, loss of Tp53 activity in cooperation with Egr1 and Apc haploinsufficiency creates an environment that is permissive for malignant transformation and the development of AML.
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Deleción Cromosómica , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Genes APC , Genes p53/fisiología , Haploinsuficiencia , Leucemia Mieloide Aguda/genética , Animales , Transformación Celular Neoplásica/genética , Células Cultivadas , Eliminación de Gen , Humanos , Ratones , Ratones Endogámicos C57BL , Ratones TransgénicosRESUMEN
The drivers of abnormal DNA methylation in human cancers include widespread aberrant splicing of the DNMT3B gene, producing abnormal transcripts that encode truncated proteins that may act as dominant negative isoforms. To test whether reduced Dnmt3b dosage can alter tumorigenesis, we bred Dnmt3b(+/-) mice to Eµ-Myc mice, a mouse model susceptible to B-cell lymphomas. Eµ-Myc/Dnmt3b(+/-) mice showed a dramatic acceleration of lymphomagenesis, greater even than that observed in Eµ-Myc mice that express a truncated DNMT3B isoform found in human tumors, DNMT3B7. This finding indicates that Dnmt3b can act as a haploinsufficient tumor suppressor gene. Although reduction in both Dnmt3b dosage and expression of DNMT3B7 within the Eµ-Myc system had similar effects on tumorigenesis and DNA hypermethylation, different molecular mechanisms appear to underlie these changes. This study offers insight into how de novo DNA methyltransferases function as tumor suppressors and the sensitivity of Myc-induced lymphomas to DNA methylation.
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Transformación Celular Neoplásica/genética , ADN (Citosina-5-)-Metiltransferasas/fisiología , Genes Supresores de Tumor/fisiología , Haploinsuficiencia/fisiología , Linfoma de Células B/genética , Proteínas Proto-Oncogénicas c-myc/fisiología , Animales , ADN (Citosina-5-)-Metiltransferasas/genética , ADN (Citosina-5-)-Metiltransferasas/metabolismo , Metilación de ADN/genética , Haploinsuficiencia/genética , Linfoma de Células B/patología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Modelos Biológicos , Regiones Promotoras Genéticas/fisiología , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , ADN Metiltransferasa 3BRESUMEN
OBJECTIVE: This study aims to evaluate ChatGPT's performance in addressing real-world otolaryngology patient questions, focusing on accuracy, comprehensiveness, and patient safety, to assess its suitability for integration into healthcare. METHODS: A cross-sectional study was conducted using patient questions from the public online forum Reddit's r/AskDocs, where medical advice is sought from healthcare professionals. Patient questions were input into ChatGPT (GPT-3.5), and responses were reviewed by 5 board-certified otolaryngologists. The evaluation criteria included difficulty, accuracy, comprehensiveness, and bedside manner/empathy. Statistical analysis explored the relationship between patient question characteristics and ChatGPT response scores. Potentially dangerous responses were also identified. RESULTS: Patient questions averaged 224.93 words, while ChatGPT responses were longer at 414.93 words. The accuracy scores for ChatGPT responses were 3.76/5, comprehensiveness scores were 3.59/5, and bedside manner/empathy scores were 4.28/5. Longer patient questions did not correlate with higher response ratings. However, longer ChatGPT responses scored higher in bedside manner/empathy. Higher question difficulty correlated with lower comprehensiveness. Five responses were flagged as potentially dangerous. CONCLUSION: While ChatGPT exhibits promise in addressing otolaryngology patient questions, this study demonstrates its limitations, particularly in accuracy and comprehensiveness. The identification of potentially dangerous responses underscores the need for a cautious approach to AI in medical advice. Responsible integration of AI into healthcare necessitates thorough assessments of model performance and ethical considerations for patient safety.
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Otolaringología , Humanos , Estudios Transversales , Relaciones Médico-Paciente/ética , Seguridad del PacienteRESUMEN
Therapy-related myeloid neoplasms (t-MNs) share many clinical and molecular characteristics with AML de novo in the elderly. One common factor is that they arise in the setting of chronic inflammation, likely because of advanced age or chemotherapy-induced senescence. Here, we examined the effect of haploinsufficient loss of the del(5q) tumor suppressor gene, EGR1, commonly deleted in high-risk MNs. In mice, under the exogenous stress of either serial transplant or successive doses of the alkylating agent N-ethyl-nitrosourea (ENU), Egr1-haploinsufficient hematopoietic stem cells (HSCs) exhibit a clonal advantage. Complete loss of EGR1 function is incompatible with transformation; mutations of EGR1 are rare and are not observed in the remaining allele in del(5q) patients, and complete knockout of Egr1 in mice leads to HSC exhaustion. Using chromatin immunoprecipitation sequencing (ChIP-seq), we identified EGR1 binding sites in human CD34+ cord blood-derived stem and progenitor cells (HSPCs) and found that EGR1 binds genes critical for stem cell differentiation, inflammatory signaling, and the DNA damage response. Notably, in the chromosome 5 sequences frequently deleted in patients, there is a significant enrichment of innate and inflammatory genes, which may confer a fitness advantage in an inflammatory environment. Short hairpin RNA (shRNA)-mediated silencing of EGR1 biases HSPCs toward a self-renewal transcriptional signature. In the absence of EGR1, HSPCs are characterized by upregulated MYC-driven proliferative signals, downregulated CDKN1A (p21), disrupted DNA damage response, and downregulated inflammation-adaptations anticipated to confer a relative fitness advantage for stem cells especially in an environment of chronic inflammation.
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Haploinsuficiencia , Células Madre Hematopoyéticas , Humanos , Ratones , Animales , Anciano , Células Madre Hematopoyéticas/metabolismo , Antígenos CD34/metabolismo , Etilnitrosourea/metabolismo , Inflamación/metabolismo , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Proteína 1 de la Respuesta de Crecimiento Precoz/metabolismoRESUMEN
Importance: Understanding risk factors for hospitalization in vaccinated persons and the association of COVID-19 vaccines with hospitalization rates is critical for public health efforts to control COVID-19. Objective: To determine characteristics of COVID-19-associated hospitalizations among vaccinated persons and comparative hospitalization rates in unvaccinated and vaccinated persons. Design, Setting, and Participants: From January 1, 2021, to April 30, 2022, patients 18 years or older with laboratory-confirmed SARS-CoV-2 infection were identified from more than 250 hospitals in the population-based COVID-19-Associated Hospitalization Surveillance Network. State immunization information system data were linked to cases, and the vaccination coverage data of the defined catchment population were used to compare hospitalization rates in unvaccinated and vaccinated individuals. Vaccinated and unvaccinated patient characteristics were compared in a representative sample with detailed medical record review; unweighted case counts and weighted percentages were calculated. Exposures: Laboratory-confirmed COVID-19-associated hospitalization, defined as a positive SARS-CoV-2 test result within 14 days before or during hospitalization. Main Outcomes and Measures: COVID-19-associated hospitalization rates among vaccinated vs unvaccinated persons and factors associated with COVID-19-associated hospitalization in vaccinated persons were assessed. Results: Using representative data from 192â¯509 hospitalizations (see Table 1 for demographic information), monthly COVID-19-associated hospitalization rates ranged from 3.5 times to 17.7 times higher in unvaccinated persons than vaccinated persons regardless of booster dose status. From January to April 2022, when the Omicron variant was predominant, hospitalization rates were 10.5 times higher in unvaccinated persons and 2.5 times higher in vaccinated persons with no booster dose, respectively, compared with those who had received a booster dose. Among sampled cases, vaccinated hospitalized patients with COVID-19 were older than those who were unvaccinated (median [IQR] age, 70 [58-80] years vs 58 [46-70] years, respectively; P < .001) and more likely to have 3 or more underlying medical conditions (1926 [77.8%] vs 4124 [51.6%], respectively; P < .001). Conclusions and Relevance: In this cross-sectional study of US adults hospitalized with COVID-19, unvaccinated adults were more likely to be hospitalized compared with vaccinated adults; hospitalization rates were lowest in those who had received a booster dose. Hospitalized vaccinated persons were older and more likely to have 3 or more underlying medical conditions and be long-term care facility residents compared with hospitalized unvaccinated persons. The study results suggest that clinicians and public health practitioners should continue to promote vaccination with all recommended doses for eligible persons.
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COVID-19 , Vacunas contra la Influenza , Adulto , Anciano , COVID-19/epidemiología , COVID-19/prevención & control , Vacunas contra la COVID-19 , Estudios Transversales , Hospitalización , Humanos , SARS-CoV-2RESUMEN
Common fragile sites (CFSs) represent large, highly unstable regions of the human genome. CFS sequences are sensitive to perturbation of replication; however, the molecular basis for the instability at CFSs is poorly understood. We hypothesized that a unique epigenetic pattern may underlie the unusual sensitivity of CFSs to replication interference. To examine this hypothesis, we analyzed chromatin modification patterns within the six human CFSs with the highest levels of breakage, and their surrounding non-fragile regions (NCFSs). Chromatin at most of the CFSs analyzed has significantly less histone acetylation than that of their surrounding NCFSs. Trichostatin A and/or 5-azadeoxycytidine treatment reduced chromosome breakage at CFSs. Furthermore, chromatin at the most commonly expressed CFS, the FRA3B, is more resistant to micrococcal nuclease than that of the flanking non-fragile sequences. These results demonstrate that histone hypoacetylation is a characteristic epigenetic pattern of CFSs, and chromatin within CFSs might be relatively more compact than that of the NCFSs, indicating a role for chromatin conformation in genomic instability at CFSs. Moreover, lack of histone acetylation at CFSs may contribute to the defective response to replication stress characteristic of CFSs, leading to the genetic instability characteristic of this regions.
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Sitios Frágiles del Cromosoma , Genoma Humano , Histonas/metabolismo , Acetilación , Línea Celular , Cromatina/genética , Cromatina/metabolismo , Epigénesis Genética , Femenino , Inestabilidad Genómica , Histonas/genética , HumanosRESUMEN
Investigations into medical student study strategies have seen an increase in recent years, but we have also seen a move to more integrated medical curricula during this time. This manuscript endeavors to assess the changes in study plans and students' reported study strategies that are associated with a move from a traditional stand-alone anatomy curriculum to an integrated, standardized curriculum. Previously validated study strategy surveys were given to medical students at the beginning of their anatomy course and again at the end of the course. These responses were then correlated with basic demographic information and outcomes in anatomy. Results indicate that this change in curriculum does correlate with changes to students' study plans and reported study strategies. In particular, the plans for and use of web-based resources appear higher in the new curriculum while the use of self-quizzing and attendance appear lower, with potentially negative implications for understanding and long-term retention. Differences were also seen between genders and student ages. Finally, a few associations with outcomes are also noted for increased use of web-based resources and student confidence going into the exam.
RESUMEN
Therapy-related myeloid neoplasms (t-MNs) following treatment with alkylating agents are characterized by a del(5q), complex karyotypes, alterations of TP53, and a dismal prognosis. To decipher the molecular pathway(s) leading to the pathogenesis of del(5q) t-MN and the effect(s) of cytotoxic therapy on the marrow microenvironment, we developed a mouse model with loss of two key del(5q) genes, EGR1 and APC, in hematopoietic cells. We used the well-characterized drug, N-ethyl-N-nitrosurea (ENU) to demonstrate that alkylating agent exposure of stromal cells in the microenvironment increases the incidence of myeloid disease. In addition, loss of Trp53 with Egr1 and Apc was required to drive the development of a transplantable leukemia, and accompanied by the acquisition of somatic mutations in DNA damage response genes. ENU treatment of mesenchymal stromal cells induced cellular senescence, and led to the acquisition of a senescence-associated secretory phenotype, which may be a critical microenvironmental alteration in the pathogenesis of myeloid neoplasms.
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Antineoplásicos Alquilantes , Médula Ósea , Leucemia Mieloide , Neoplasias Primarias Secundarias , Animales , Antineoplásicos Alquilantes/efectos adversos , Antineoplásicos Alquilantes/uso terapéutico , Deleción Cromosómica , Genes p53 , Leucemia Mieloide/inducido químicamente , Leucemia Mieloide/genética , Ratones , Neoplasias Primarias Secundarias/inducido químicamente , Neoplasias Primarias Secundarias/genética , Células del Estroma , Microambiente Tumoral/genéticaRESUMEN
This study explores the construction of risk and patient identity in medical discourse directed toward women with breast cancer. Eleven documents produced by the National Cancer Institute on the topic of breast cancer are studied using narrative analysis. A distinct patient narrative presents all women as at risk for breast cancer and creates an idealized patient identity that serves a prescriptive function for women. The narrative constructs an early-cancer experience where the patient is treatable and cancer is cured or controlled. There are no significant changes in the narrative after time.
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Neoplasias de la Mama/psicología , Medición de Riesgo , Autoimagen , Actitud Frente a la Salud , Imagen Corporal , Mama , Características Culturales , Femenino , Humanos , Folletos , Investigación CualitativaRESUMEN
The FRA3B, at 3p14.2, lies within the fragile histidine triad (FHIT) gene and is the most highly expressed of the common fragile sites observed when DNA replication is perturbed by aphidicolin. Common fragile sites are highly unstable regions of the genome. Large intragenic deletions within FHIT, localized within the FRA3B sequences, have been identified in a variety of tumor cells. To characterize the FRA3B deletions in tumor cells and identify FRA3B sequences that are required for fragile site induction, we used microcell-mediated chromosome transfer to isolate hybrid cell clones that retain chromosome 3 homologues with various deletions within FRA3B. Detailed molecular mapping of the FHIT/FRA3B locus in the resultant hybrid cells revealed a complex pattern of instability within FRA3B. Each tumor cell line contained multiple chromosome 3 homologues with variable deletion patterns, often with discontinuous deletions, suggesting that the process of breakage and repair within FRA3B is an ongoing one. By comparing the approximate location of the breakpoints in the hybrid clones, we identified 11 recurring breakpoint/repair regions within the FRA3B. A comparison of the frequency of breaks/gaps within FRA3B in the hybrid clones with various deletions of FRA3B sequences revealed that the loss of FRA3B sequences does not reduce the overall rate of breakage and instability within the remaining FRA3B sequences. The majority of breaks occurred in the proximal portion of the FRA3B, in a 300-kb interval between exon 4 and the proximal 50 kb of intron 5. Our observations suggest that there is no single sequence within the FRA3B that influences breakage or recombination within this region; however, we cannot rule out the presence of multiple "hot spots" within the FHIT/FRA3B locus. Together, the results suggest that factors other than the DNA sequence per se are responsible for the formation of DNA breaks/gaps.
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Ácido Anhídrido Hidrolasas , Fragilidad Cromosómica/genética , Cromosomas Humanos Par 3/genética , Proteínas de Neoplasias/genética , Neoplasias/genética , Animales , Células CHO , Carcinoma de Pulmón de Células no Pequeñas/genética , Sitios Frágiles del Cromosoma , Mapeo Cromosómico , Cricetinae , Femenino , Eliminación de Gen , Técnicas de Transferencia de Gen , Humanos , Neoplasias Pulmonares/genética , Neoplasias Gástricas/genética , Células Tumorales Cultivadas , Neoplasias del Cuello Uterino/genéticaRESUMEN
Chromosomal rearrangements involving the immunoglobulin heavy chain gene (IGH) at 14q32 are observed in approximately 50% of patients with B-cell non-Hodgkin's lymphoma (NHL). The 5' end of the IGH gene is located within 8 kb of the telomeric repeats of 14q. Translocations involving the IGH locus and the telomeric band of a partner chromosome are difficult to identify, because most terminal bands of human chromosomes appear pale by conventional G-banding techniques. To determine whether there are cryptic translocations involving the IGH locus, we used dual-color fluorescence in situ hybridization (FISH) of 5' and 3' IGH genomic clones containing the variable sequences, or the J(H) and the 5' constant regions, respectively. We examined cells from 51 patients with B-cell NHL who had a normal karyotype (3 patients), clonal abnormalities not involving 14q32 (35 patients), or alterations of 14q32 other than recurring translocations, i.e., add(14)(q32) (13 patients). FISH detected 17 IGH translocations in 16 of 51 (31%) cases. Of the 13 cases with add(14)(q32), FISH identified the partner chromosome in 9 cases (69%; 3q27, 6 cases; 2p13, 19p13.3, and 18q21.3, 1 case each). Six of thirty-eight (16%) patients without visible alterations of 14q32 and 2 of 13 (15%) patients with an abnormality of one chromosome 14 had masked (5 patients) or cryptic IGH translocations (3 patients), involving 3q27 (3 patients), 5p15.3 (2 patients), 19p13.3 (3 patients), or 14q32 (1 patient; 1 patient had two rearrangements). We identified two novel, recurring, cryptic translocations: t(5;14)(p15.3;q32) (2 patients) and t(14;19)(q32;p13.3) (3 patients). In summary, FISH permitted the detection of cryptic or masked IGH rearrangements in approximately 20% of lymphoma cases without visible rearrangements of 14q32 analyzed retrospectively.
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Genes de Inmunoglobulinas/genética , Linfoma de Células B/genética , Adulto , Anciano , Anciano de 80 o más Años , Preescolar , Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 5/genética , Femenino , Humanos , Hibridación Fluorescente in Situ , Linfoma de Células B/inmunología , Linfoma de Células B/patología , Masculino , Persona de Mediana Edad , Translocación GenéticaRESUMEN
BACKGROUND: Angioedema of the upper aerodigestive tract can lead to significant airway obstruction. To date no articles have delineated risk factors for progression after initial evaluation. METHODS: This article presents the results of a retrospective study of patients with angioedema at a single institution. Patients included were consecutive otolaryngology consultations for angioedema in the emergency department (ED) from 1999 to 2003. All patients were evaluated by an otolaryngologist and underwent fiber-optic laryngoscopy, which was repeated serially depending on findings. Data was collected on demographics, comorbidities, intubation, disposition, and progression of angioedema. RESULTS: A total of 177 patients were included in the study: 32 (18%) patients required intubation; 25 (14%) on initial presentation and 7 (4%) who progressed from an initially stable airway to requiring intervention after reevaluation. Analysis of variance (ANOVA) demonstrated a statistically significant variance between location of edema and rate of intubation, with higher rates in the pharynx and larynx vs the lip and face. Patients who required intubation after progression between serial evaluations were statistically more likely to have edema that involved deeper portions of the aerodigestive tract. Patients who required intubation were statistically more likely to be older (average age 61.8 vs 55.1 years, p = 0.03). CONCLUSION: In this large series of patients managed for aerodigestive angioedema we demonstrate risk factors associated with airway intervention, and risk factors associated with clinical progression on serial examination to airway intervention. In addition, we demonstrate a successful management algorithm for patients with aerodigestive angioedema.
Asunto(s)
Factores de Edad , Obstrucción de las Vías Aéreas/epidemiología , Obstrucción de las Vías Aéreas/terapia , Angioedema/epidemiología , Angioedema/terapia , Intubación Intratraqueal , Sistema Respiratorio/metabolismo , Obstrucción de las Vías Aéreas/etiología , Algoritmos , Angioedema/complicaciones , Comorbilidad , Servicio de Urgencia en Hospital , Femenino , Humanos , Laringoscopía , Masculino , Persona de Mediana Edad , Sistema Respiratorio/patología , Estudios Retrospectivos , Factores de RiesgoRESUMEN
OBJECTIVES/HYPOTHESIS: Within otolaryngology residency training, the annual Otolaryngology Training Examination (OTE) is the primary method used to assess, quantify, and compare the factual knowledge acquired by each resident. The objective of this study was to develop a more frequent method for tracking of factual knowledge to prevent educational delay. STUDY DESIGN: Retrospective analysis of educational scores. METHODS: For each didactic lecture within a single otolaryngology residency training program, multiple choice questions were provided before and after each lecture. Questions were based on lecture objectives that were derived from the American Board of Otolaryngology curriculum. Scores were tracked over the course of 1 academic year and compared to the scores of residents on the OTE administered in that year to determine correlation with a validated measure of factual knowledge. The effect of remedial measures on improvement in OTE scores was determined. RESULTS: Over the course of 1 academic year, there were 328 questions presented to 12 residents before and after 32 lectures in the didactic program. Ten residents completed an average of 244 questions. Overall OTE scores demonstrated a significant and very strong correlation to lecture question scores (Pearson r = 0.86, P = .002). Remedial measures for residents during the previous 5 years who had inadequate OTE scores were successful in improving scores (P = .002). CONCLUSIONS: A structured didactic program that uses review questions to assess knowledge can be used to track acquisition of factual knowledge. Early identification of residents with deficiencies facilitates the development of individualized learning plans that result in successful remediation.
Asunto(s)
Competencia Clínica , Internado y Residencia , Otolaringología/educación , Estudios Retrospectivos , Factores de TiempoRESUMEN
Therapy-related myeloid neoplasm (t-MN) is a distinctive clinical syndrome occurring after exposure to chemotherapy or radiotherapy. t-MN arises in most cases from a multipotential hematopoietic stem cell or, less commonly, in a lineage committed progenitor cell. The prognosis for patients with t-MN is poor, as current forms of therapy are largely ineffective. Cytogenetic analysis, molecular analysis and gene expression profiling analysis of t-MN has revealed that there are distinct subtypes of the disease; however, our understanding of the genetic basis of t-MN is incomplete. Elucidating the genetic pathways and molecular networks that are perturbed in t-MNs, may facilitate the identification of therapeutic targets that can be exploited for the development of urgently-needed targeted therapies.
RESUMEN
Epigenetic changes are among the most common alterations observed in cancer cells, yet the mechanism by which cancer cells acquire and maintain abnormal DNA methylation patterns is not understood. Cancer cells have an altered distribution of DNA methylation and express aberrant DNA methyltransferase 3B transcripts, which encode truncated proteins, some of which lack the COOH-terminal catalytic domain. To test if a truncated DNMT3B isoform disrupts DNA methylation in vivo, we constructed two lines of transgenic mice expressing DNMT3B7, a truncated DNMT3B isoform commonly found in cancer cells. DNMT3B7 transgenic mice exhibit altered embryonic development, including lymphopenia, craniofacial abnormalities, and cardiac defects, similar to Dnmt3b-deficient animals, but rarely develop cancer. However, when DNMT3B7 transgenic mice are bred with Emicro-Myc transgenic mice, which model aggressive B-cell lymphoma, DNMT3B7 expression increases the frequency of mediastinal lymphomas in Emicro-Myc animals. Emicro-Myc/DNMT3B7 mediastinal lymphomas have more chromosomal rearrangements, increased global DNA methylation levels, and more locus-specific perturbations in DNA methylation patterns compared with Emicro-Myc lymphomas. These data represent the first in vivo modeling of cancer-associated DNA methylation changes and suggest that truncated DNMT3B isoforms contribute to the redistribution of DNA methylation characterizing virtually every human tumor.