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1.
Commun Med (Lond) ; 2: 86, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35865358

RESUMEN

Easy access to large quantities of accurate health data is required to understand medical and scientific information in real-time; evaluate public health measures before, during, and after times of crisis; and prevent medical errors. Introducing a system in the USA that allows for efficient access to such health data and ensures auditability of data facts, while avoiding data silos, will require fundamental changes in current practices. Here, we recommend the implementation of standardized data collection and transmission systems, universal identifiers for individual patients and end users, a reference standard infrastructure to support calibration and integration of laboratory results from equivalent tests, and modernized working practices. Requiring comprehensive and binding standards, rather than incentivizing voluntary and often piecemeal efforts for data exchange, will allow us to achieve the analytical information environment that patients need.

2.
Am J Med ; 130(7): 864-866, 2017 07.
Artículo en Inglés | MEDLINE | ID: mdl-28344147

RESUMEN

BACKGROUND: Recent studies have shed light on the continued prescription of inpatient medications upon hospital discharge, despite the original intent of short-term inpatient therapy. Amiodarone, an antiarrhythmic associated with significant adverse effects with long-term use, is commonly used for new-onset atrial fibrillation in critical illness (NAFCI). Although it is often preferred in this setting of hemodynamic instability, a prescription for long-term use should be carefully considered, preferably by a cardiologist. This study was conducted to evaluate the incidence of patients discharged on amiodarone without a cardiology consult or referral after being initiated on amiodarone for NAFCI. METHODS: We conducted a retrospective review of all patients newly prescribed amiodarone for NAFCI over a 2-year period. The primary outcome was the percentage of patients who were continued on amiodarone upon hospital discharge without review by or outpatient referral to a cardiologist. RESULTS: Of the 100 patients who met inclusion criteria, 59 patients were prescribed amiodarone upon hospital discharge. Of these, 48 patients (81.4%) had converted to normal sinus rhythm with the resolution of critical illness. Of 100 patients, 23 received prescriptions for amiodarone upon discharge without a cardiology consult or referral. CONCLUSION: Amiodarone was frequently continued upon discharge without referral to a cardiologist in patients initiated on this therapy for NAFCI. This may contribute to unnecessary long-term therapy, thereby increasing the risk for significant side effects, drug interactions, and increased healthcare costs. This study suggests that careful medication reconciliation through all transitions of care, including discharge, is essential.


Asunto(s)
Amiodarona/uso terapéutico , Antiarrítmicos/uso terapéutico , Fibrilación Atrial/tratamiento farmacológico , Enfermedad Crítica , Deprescripciones , Alta del Paciente , Anciano , Amiodarona/efectos adversos , Amiodarona/economía , Antiarrítmicos/efectos adversos , Antiarrítmicos/economía , Cardiólogos , Esquema de Medicación , Costos de los Medicamentos , Femenino , Humanos , Masculino , Conciliación de Medicamentos , Derivación y Consulta , Estudios Retrospectivos
3.
Ann Am Thorac Soc ; 14(3): 368-375, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-27983871

RESUMEN

RATIONALE: Video laryngoscopy has overcome the need to align the anatomic axes to obtain a view of the glottic opening to place a tracheal tube. However, despite this advantage, a large number of attempts are unsuccessful. There are no existing data on anatomic characteristics in critically ill patients associated with a failed first attempt at laryngoscopy when using video laryngoscopy. OBJECTIVES: To identify characteristics associated with first-attempt failure at intubation when using video laryngoscopy in the intensive care unit (ICU). METHODS: This is an observational study of 906 consecutive patients intubated in the ICU with a video laryngoscope between January 2012 and January 2016 in a single-center academic medical ICU. After each intubation, the operator completed a data collection form, which included information on difficult airway characteristics, device used, and outcome of each attempt. Multivariable regression models were constructed to determine the difficult airway characteristics associated with a failed first attempt at intubation. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in sex, age, reason for intubation, or device used between first-attempt failures and first-attempt successes. First-attempt successes more commonly reported no difficult airway characteristics were present (23.9%; 95% confidence interval [CI], 20.7-27.0% vs. 13.3%; 95% CI, 8.0-18.8%). In logistic regression analysis of the entire 906-patient database, blood in the airway (odds ratio [OR], 2.63; 95% CI, 1.64-4.20), airway edema (OR, 2.85; 95% CI, 1.48-5.45), and obesity (OR, 1.59; 95% CI, 1.08-2.32) were significantly associated with first-attempt failure. Data collection on limited mouth opening and secretions began after the first 133 intubations, and we fit a second logistic model to examine cases in which these additional difficult airway characteristics were collected. In this subset (n = 773), the presence of blood (OR, 2.73; 95% CI, 1.60-4.64), cervical immobility (OR, 3.34; 95% CI, 1.28-8.72), and airway edema (OR, 3.10; 95% CI, 1.42-6.70) were associated with first-attempt failure. CONCLUSIONS: In this single-center study, presence of blood in the airway, airway edema, cervical immobility, and obesity are associated with higher odds of first-attempt failure, when intubation was performed with video laryngoscopy in an ICU.


Asunto(s)
Intubación Intratraqueal/métodos , Laringoscopía/métodos , Sistema Respiratorio/fisiopatología , Grabación en Video , Anciano , Arizona , Cuidados Críticos/métodos , Bases de Datos Factuales , Femenino , Humanos , Unidades de Cuidados Intensivos , Intubación Intratraqueal/instrumentación , Modelos Logísticos , Lesión Pulmonar/etiología , Masculino , Persona de Mediana Edad , Análisis Multivariante , Oportunidad Relativa , Evaluación de Procesos y Resultados en Atención de Salud
4.
Intern Emerg Med ; 12(8): 1235-1243, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27738960

RESUMEN

The purpose of this investigation was to investigate the association between first attempt success and intubation-related complications in the Intensive Care Unit after the widespread adoption of video laryngoscopy. We further sought to characterize and identify the predictors of complications that occur despite first attempt success. This was a prospective observational study of consecutive intubations performed with video laryngoscopy at an academic medical Intensive Care Unit. Operator, procedural, and complication data were collected. Multivariable logistic regression was used to examine the relationship between the intubation attempts and the occurrence of one or more complications. A total of 905 patients were intubated using a video laryngoscope. First attempt success occurred in 739 (81.7 %), whereas >1 attempt was needed in 166 (18.3 %). One or more complications occurred in 146 (19.8 %) of those intubated on the first attempt versus 107 (64.5 %, p < 0.001) of those requiring more than one attempt. Logistic regression analysis shows that >1 attempt is associated with 6.4 (95 % CI 4.4-9.3) times the adjusted odds of at least one complication. Pre-intubation predictors of at least one complication despite first attempt success include vomit or edema in the airway as well as the presence of hypoxemia or hypotension. There are increased odds of complications with even a second attempt at intubation in the Intensive Care Unit. Complications occur frequently despite a successful first attempt, and as such, the goal of airway management should not be simply first attempt success, but instead first attempt success without complications.


Asunto(s)
Intubación Intratraqueal/normas , Laringoscopía/efectos adversos , Laringoscopía/normas , Anciano , Manejo de la Vía Aérea/normas , Femenino , Humanos , Unidades de Cuidados Intensivos/organización & administración , Unidades de Cuidados Intensivos/estadística & datos numéricos , Intubación Intratraqueal/estadística & datos numéricos , Laringoscopía/instrumentación , Modelos Logísticos , Masculino , Persona de Mediana Edad , Neumonía por Aspiración/complicaciones
5.
Ann Am Thorac Soc ; 13(3): 382-90, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26653096

RESUMEN

RATIONALE: Urgent tracheal intubation is performed frequently in intensive care units and incurs higher risk than when intubation is performed under more controlled circumstances. Video laryngoscopy may improve the chances of successful tracheal intubation on the first attempt; however, existing comparative data on outcomes are limited. OBJECTIVES: To compare first-attempt success and complication rates during intubation when using video laryngoscopy compared with traditional direct laryngoscopy in a tertiary academic medical intensive care unit. METHODS: We prospectively collected and analyzed data from a continuous quality improvement database of all intubations in one medical intensive care unit between January 1, 2012, and December 31, 2014. Propensity matching and multivariable logistic regression were used to reduce the risk of bias and control for confounding. MEASUREMENTS AND MAIN RESULTS: A total of 809 intubations took place over the study period. Of these, 673 (83.2%) were performed using video laryngoscopy and 136 (16.8%) using direct laryngoscopy. First-attempt success with video laryngoscopy was 80.4% (95% confidence interval [CI], 77.2-83.3%) compared with 65.4% (95% CI, 56.8-73.4%) for intubations performed with direct laryngoscopy (P < 0.001). In a propensity-matched analysis, the odds ratio for first-attempt success with video laryngoscopy versus direct laryngoscopy was 2.81 (95% CI, 2.27-3.59). The rate of arterial oxygen desaturation events during the first intubation attempt was significantly lower for video laryngoscopy than for direct laryngoscopy (18.3% vs. 25.9%; P = 0.04). The rate of esophageal intubation during any attempt was also significantly lower for video laryngoscopy (2.1% vs. 6.6%; P = 0.008). CONCLUSIONS: Video laryngoscopy was associated with significantly improved odds of first-attempt success at tracheal intubation by nonanesthesiologists in a medical intensive care unit. Esophageal intubation and oxygen desaturation occurred less frequently with the use of video laryngoscopy. Randomized clinical trials are needed to confirm these findings.


Asunto(s)
Unidades de Cuidados Intensivos/organización & administración , Intubación Intratraqueal/métodos , Laringoscopía/métodos , Cirugía Asistida por Video/métodos , Anciano , Arizona , Femenino , Humanos , Intubación Intratraqueal/normas , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Oportunidad Relativa , Complicaciones Posoperatorias/epidemiología , Estudios Prospectivos , Mejoramiento de la Calidad , Centros de Atención Terciaria
6.
Ann Intensive Care ; 5: 4, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25852964

RESUMEN

BACKGROUND: Noninvasive positive-pressure ventilation (NIPPV) use has increased in the treatment of patients with respiratory failure. However, despite decreasing the need for intubation in some patients, there are no data regarding the risk of intubation-related complications associated with delayed intubation in adult patients who fail NIPPV. The objective of this study is to evaluate the odds of a composite complication of intubation following failed NIPPV compared to patients intubated primarily in the medical intensive care unit (ICU). METHODS: This is a single-center retrospective cohort study of 235 patients intubated between 1 January 2012 and 30 June 2013 in a medical ICU of a university medical center. A total of 125 patients were intubated after failing NIPPV, 110 patients were intubated without a trial of NIPPV. Intubation-related data were collected prospectively through a continuous quality improvement (CQI) program and retrospectively extracted from the medical record on all patients intubated on the medical ICU. A propensity adjustment for the factors expected to affect the decision to initially use NIPPV was used, and the adjusted multivariate regression analysis was performed to evaluate the odds of a composite complication (desaturation, hypotension, or aspiration) with intubation following failed NIPPV versus primary intubation. RESULTS: A propensity-adjusted multivariate regression analysis revealed that the odds of a composite complication of intubation in patients who fail NIPPV was 2.20 (CI 1.14 to 4.25), when corrected for the presence of pneumonia or acute respiratory distress syndrome (ARDS), and adjusted for factors known to increase complications of intubation (total attempts and operator experience). When a composite complication occurred, the unadjusted odds of death in the ICU were 1.79 (95% CI 1.03 to 3.12). CONCLUSIONS: After controlling for potential confounders, this propensity-adjusted analysis demonstrates an increased odds of a composite complication with intubation following failed NIPPV. Further, the presence of a composite complication during intubation is associated with an increased odds of death in the ICU.

7.
Ann Am Thorac Soc ; 12(5): 734-41, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25719512

RESUMEN

RATIONALE: The use of neuromuscular blocking agents (NMBAs) has been shown to be valuable in improving successful tracheal intubation in the operating room and emergency department. However, data on NMBA use in critically ill intensive care unit (ICU) patients are lacking. Furthermore, there are no data on NMBA use with video laryngoscopy. OBJECTIVES: To evaluate the effect of NMBA use on first-attempt success (FAS) with tracheal intubation in the ICU. METHODS: Single-center observational study of 709 consecutive patients intubated in the medical ICU of a university medical center from January 1, 2012 to June 30, 2014. Data were collected prospectively through a continuous quality improvement program on all patients intubated in the ICU over the study period. Data relating to patient demographics, intubation, and complications were analyzed. We used propensity score (propensity to use an NMBA) matching to generate 5,000 data sets of cases (failed first intubation attempts) matched to controls (successful first attempts) and conditional logistic regression to analyze the results. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in patient demographics, except median total difficult airway characteristics were higher in the non-NMBA group (2 vs. 1, P < 0.001). There were significant differences in the sedative used between groups and the operator level of training. More patients who were given NMBAs received etomidate (83 vs. 35%) and more patients in the non-NMBA group received ketamine (39 vs. 9%) (P < 0.001). The FAS for NMBA use was 80.9% (401/496) compared with 69.6% (117/168) for non-NMBA use (P = 0.003). The summary odds ratio for FAS when an NMBA was used from the propensity matched analyses was 2.37 (95% confidence interval, 1.36-4.88). In the subgroup of patients intubated with a video laryngoscope, propensity-adjusted odds of FAS with the use of an NMBA was 2.50 (1.43-4.37; P < 0.001). There were no differences in procedurally related complications between groups. CONCLUSIONS: After controlling for potential confounders, this propensity-adjusted analysis demonstrates improved odds of FAS at intubation in the ICU with the use of an NMBA. This improvement in FAS is seen even with the use of a video laryngoscope.


Asunto(s)
Manejo de la Vía Aérea/métodos , Unidades de Cuidados Intensivos , Intubación Intratraqueal/métodos , Laringoscopía/métodos , Bloqueo Neuromuscular/métodos , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Reproducibilidad de los Resultados , Grabación en Video
8.
Ann Am Thorac Soc ; 12(4): 539-48, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25715227

RESUMEN

RATIONALE: Airway management in the intensive care unit (ICU) is challenging, as many patients have limited physiologic reserve and are at risk for clinical deterioration if the airway is not quickly secured. In academic medical centers, ICU intubations are often performed by trainees, making airway management education paramount for pulmonary and critical care trainees. OBJECTIVES: To improve airway management education for our trainees, we developed a comprehensive training program including an 11-month simulation-based curriculum. The curriculum emphasizes recognition of and preparation for potentially difficult intubations and procedural skills to maximize patient safety and increase the likelihood of first-attempt success. METHODS: Training is provided in small group sessions twice monthly using a high-fidelity simulation program under the guidance of a core group of two to three advanced providers. The curriculum is designed with progressively more difficult scenarios requiring critical planning and execution of airway management by the trainees. Trainees consider patient position, preoxygenation, optimization of hemodynamics, choice of induction agents, selection of appropriate devices for the scenario, anticipation of difficulties, back-up plans, and immediate postintubation management. Clinical performance is monitored through a continuous quality improvement program. MEASUREMENTS AND MAIN RESULTS: Sixteen fellows have completed the program since July 1, 2013. In the 18 months since the start of the curriculum (July 1, 2013-December 31, 2014), first-attempt success has improved from 74% (358/487) to 82% (305/374) compared with the 18 months before implementation (P = 0.006). During that time there were no serious complications related to airway management. Desaturation rates decreased from 26 to 17% (P = 0.002). Other complication rates are low, including aspiration (2.1%), esophageal intubation (2.7%), dental trauma (0.8%), and hypotension (8.3%). First-attempt success in a 6-month period after implementation (July 1, 2014-December 31, 2014) was significantly higher (82.1 compared with 70.9%, P = 0.03) than during a similar 6-month period before implementation (July 1, 2012-December 31, 2012). CONCLUSIONS: This comprehensive airway curriculum is associated with improved first-attempt success rate for intensive care unit intubations. Such a curriculum holds the potential to improve patient care.


Asunto(s)
Competencia Clínica , Cuidados Críticos , Becas/métodos , Intubación Intratraqueal , Laringoscopía/educación , Neumología/educación , Entrenamiento Simulado/métodos , Anciano , Manejo de la Vía Aérea , Curriculum , Femenino , Humanos , Masculino , Persona de Mediana Edad
9.
J Crit Care ; 29(4): 645-9, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24768532

RESUMEN

PURPOSE: The purpose of this study is to describe the current state of bedside ultrasound use and training among critical care (CC) training programs in the United States. MATERIALS AND METHODS: This was a cross-sectional survey of all program directors for Accreditation Council for Graduate Medical Education accredited programs during the 2012 to 2013 academic year in CC medicine, surgical CC, pulmonary and critical care, and anesthesia CC. Availability, current use, and barriers to training in CC ultrasound were assessed. RESULTS: Sixty of 195 (31%; 95% confidence interval [CI], 24%-38%) program directors responded. Most of the responding programs had an ultrasound system available for use (54/60, 90%; 95% CI, 79%-96%) and identified ultrasound training as useful (59/60, 98%; 95% CI, 91%-100%) but lacked a formal curriculum (25/60, 42%; 95% CI, 29%-55%) or trained faculty (mean percentage of faculty trained in ultrasound: pulmonary and critical care, 25%; surgical CC, 33%; anesthesia CC, 20%; CC medicine, 7%), and relied on informal teaching (45/60, 77%; 95% CI, 62%-85%). Faculty with expertise (53/60, 88%; 95% CI, 77%-95%), simulation training (60/60, 100%; 95% CI, 94%-100%), establishing and meeting required number of examinations (47/60, 78%; 95% CI, 66%-88%), and regular review sessions (49/60, 82%; 95% CI, 70%-90%) were identified as necessary to improve ultrasound training. Most responding programs (32/35 91%; 95% CI, 77%-98%) without a formal curriculum plan to create one in the next 5 years. CONCLUSIONS: This study identified deficiencies in current training, suggesting a need for a formal curriculum for bedside ultrasound training in CC fellowship programs.


Asunto(s)
Cuidados Críticos , Sistemas de Atención de Punto , Ultrasonido/educación , Acreditación , Estudios Transversales , Recolección de Datos , Docentes Médicos/provisión & distribución , Becas , Femenino , Humanos , Ultrasonido/organización & administración , Ultrasonido/estadística & datos numéricos , Estados Unidos
10.
Crit Care ; 17(5): R237, 2013 Oct 14.
Artículo en Inglés | MEDLINE | ID: mdl-24125064

RESUMEN

INTRODUCTION: Tracheal intubation in the Intensive Care Unit (ICU) can be challenging as patients often have anatomic and physiologic characteristics that make intubation particularly difficult. Video laryngoscopy (VL) has been shown to improve first attempt success compared to direct laryngoscopy (DL) in many clinical settings and may be an option for ICU intubations. METHODS: All intubations performed in this academic medical ICU during a 13-month period were entered into a prospectively collected quality control database. After each intubation, the operator completed a standardized form evaluating multiple aspects of the intubation including: patient demographics, difficult airway characteristics (DACs), method and device(s) used, medications used, outcomes and complications of each attempt. Primary outcome was first attempt success. Secondary outcomes were grade of laryngoscopic view, ultimate success, esophageal intubations, and desaturation. Multivariate logistic regression was performed for first attempt and ultimate success. RESULTS: Over the 13-month study period (January 2012-February 2013), a total of 234 patients were intubated using VL and 56 patients were intubated with DL. First attempt success for VL was 184/234 (78.6%; 95% CI 72.8 to 83.7) while DL was 34/56 patients (60.7%; 95% CI 46.8 to 73.5). Ultimate success for VL was 230/234 (98.3%; 95% CI 95.1 to 99.3) while DL was 52/56 patients (91.2%; 95% CI 81.3 to 97.2). In the multivariate regression model, VL was predictive of first attempt success with an odds ratio of 7.67 (95% CI 3.18 to 18.45). VL was predictive of ultimate success with an odds ratio of 15.77 (95% CI 1.92 to 129). Cormack-Lehane I or II view occurred 199/234 times (85.8%; 95% CI 79.5 to 89.1) and a median POGO (Percentage of Glottic Opening) of 82% (IQR 60 to 100) with VL, while Cormack-Lehane I or II view occurred 34/56 times (61.8%; 95% CI 45.7 to 71.9) and a median POGO of 45% (IQR 0 to 78%) with DL. VL reduced the esophageal intubation rate from 12.5% with DL to 1.3% (P = 0.001) but there was no difference in desaturation rates. CONCLUSIONS: In the medical ICU, video laryngoscopy resulted in higher first attempt and ultimate intubation success rates and improved grade of laryngoscopic view while reducing the esophageal intubation rate compared to direct laryngoscopy.


Asunto(s)
Unidades de Cuidados Intensivos , Intubación Intratraqueal/métodos , Laringoscopía/métodos , Grabación en Video , Esófago , Humanos , Resultado del Tratamiento
11.
Am J Crit Care ; 21(4): 280-6, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22751371

RESUMEN

BACKGROUND: Albumin is broadly prescribed for critically ill patients although it does not have a mortality benefit over crystalloids. One common use of albumin is to promote diuresis. Objectives To compare urine output in patients treated with furosemide with and without albumin and to assess other variables possibly associated with enhanced diuresis. METHODS: A retrospective study was conducted on patients in a medical intensive care unit who received furosemide therapy as a continuous infusion with and without 25% albumin for more than 6 hours. Primary end points were urine output and net fluid loss. RESULTS: A total of 31 patients were included in the final analysis. Mean urine output in patients treated with furosemide alone did not differ significantly from output in patients treated with furo-semide plus albumin at 6, 24, and 48 hours: mean output, 1119 (SD, 597) mL vs 1201 (SD, 612) mL, P = .56; 4323 (SD, 1717) mL vs 4615 (SD, 1741) mL, P = .42; and 7563 mL (SD, 2766) vs 7432 (SD, 2324) mL, P = .94, respectively. Additionally, net fluid loss did not differ significantly between the 2 groups at 6, 24, and 48 hours. Higher concentrations of serum albumin did not improve urine output. The only independent variable significantly associated with enhanced urine output at 24 and 48 hours was increased fluid intake. CONCLUSION: Addition of albumin to a furosemide infusion did not enhance diuresis obtained with furosemide alone in critically ill patients.


Asunto(s)
Diuresis/efectos de los fármacos , Furosemida/administración & dosificación , Hipoalbuminemia/tratamiento farmacológico , Albúmina Sérica/administración & dosificación , Adulto , Anciano , Anciano de 80 o más Años , Arizona , Cuidados Críticos/métodos , Diuréticos/administración & dosificación , Quimioterapia Combinada , Femenino , Humanos , Modelos Lineales , Masculino , Persona de Mediana Edad , Evaluación de Resultado en la Atención de Salud , Estudios Retrospectivos , Albúmina Sérica/farmacología , Adulto Joven
12.
Manag Care Interface ; 20(8): 29-34, 2007 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18041348

RESUMEN

A continuing goal of treatment of diabetes is to slow the progression of the disease through the preservation of beta-cell function. To assess efficacious and cost-effective methods of doing this, The Pharmacy & Therapeutics Society (Glastonbury, CT) assembled a study group of health plan medical directors. Based on a review of the data, the group studied the value of earlier diagnosis and the administration of thiazolidinediones. The potential role for health plans to support this approach to diabetes care is outlined below.


Asunto(s)
Diabetes Mellitus/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Células Secretoras de Insulina/efectos de los fármacos , Atención Primaria de Salud/normas , Tiazolidinedionas/uso terapéutico , Análisis Costo-Beneficio , Diabetes Mellitus/metabolismo , Progresión de la Enfermedad , Diagnóstico Precoz , Política de Salud , Humanos , Hipoglucemiantes/economía , Células Secretoras de Insulina/metabolismo , Programas Controlados de Atención en Salud/economía , Atención Primaria de Salud/economía , Calidad de Vida , Tiazolidinedionas/economía
14.
Mol Endocrinol ; 17(12): 2566-82, 2003 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-12920235

RESUMEN

We developed a molecular genetic model to investigate glucocorticoid receptor (GR) signaling in human bronchial epithelial cells in response to the therapeutic steroid budesonide. Based on a genetic selection scheme using the human Chago K1 cell line and integrated copies of a glucocorticoid-responsive herpes simplex virus thymidine kinase gene and a green fluorescent protein gene, we isolated five Chago K1 variants that grew in media containing budesonide and ganciclovir. Three spontaneous budesonide-resistant subclones were found to express low levels of GR, whereas two mutants isolated from ethylmethane sulfonate-treated cultures contained normal levels of GR protein. Analysis of the GR coding sequence in the budesonide-resistant subclone Ch-BdE5 identified a novel Val to Met mutation at amino acid position 575 (GRV575M) which caused an 80% decrease in transcriptional regulatory functions with only a minimal effect on ligand binding activity. Homology modeling of the GR structure in this region of the hormone binding domain and molecular dynamic simulations suggested that the GRV575M mutation would have a decreased affinity for the LXXLL motif of p160 coactivators. To test this prediction, we performed transactivation and glutathione-S-transferase pull-down assays using the p160 coactivator glucocorticoid interacting protein 1 (GRIP1)/transcriptional intermediary factor 2 and found that GRV575M transcriptional activity was not enhanced by GRIP1 in transfected cells nor was it able to bind GRIP1 in vitro. Identification of the novel GRV575M variant in human bronchial epithelial cells using a molecular genetic selection scheme suggests that functional assays performed in relevant cell types could identify subtle defects in GR signaling that contribute to reduced steroid sensitivities in vivo.


Asunto(s)
Bronquios/fisiología , Budesonida/farmacología , Receptores de Glucocorticoides/genética , Mucosa Respiratoria/fisiología , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Secuencia de Bases , Sitios de Unión , Carcinoma Broncogénico , Línea Celular , Línea Celular Tumoral , Resistencia a Medicamentos , Ganciclovir/farmacología , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Humanos , Neoplasias Pulmonares , Metionina , Datos de Secuencia Molecular , Membrana Mucosa , Mutación Missense , Proteínas Recombinantes de Fusión/metabolismo , Mucosa Respiratoria/efectos de los fármacos , Transfección , Valina
15.
J Steroid Biochem Mol Biol ; 84(4): 441-52, 2003 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-12732289

RESUMEN

Molecular analysis of steroid-regulated gene expression in freshly isolated human eosinophils is difficult due to the inherent high rate of spontaneous apoptosis and elevated levels of endogenous ribonucleases. To circumvent these limitations, we determined if the human eosinophilic cell line EoL-1 could serve as an in vitro model of glucocorticoid signaling. We found by optimizing growth conditions in low serum-containing media that dexamethasone (Dex) treatment of EoL-1 cells induced an apoptotic pathway that was inhibited by interleukin-5 (IL-5). Moreover, gene expression profiling using RNA from untreated EoL-1 cells and from freshly isolated human eosinophils identified 380 commonly expressed genes, including the eosinophil markers granule major basic protein, prostaglandin-endoperoxide synthase 1 and arachidonate 15-lipoxygenase. Expression profiling was performed using EoL-1 cells that had been treated with dexamethasone for 0, 4, 12, 24 and 48h identifying 162 genes as differentially expressed. Two of the most highly upregulated genes based on expression profiling were the transcription factor Ets-2 and the MHC Class II genes (Q, R, and P). Expression of these genes in EoL-1 cells was shown to be dexamethasone-induced at the RNA and protein levels which is consistent with the known function of Ets-2 in controlling cell cycle progression and the role of MHC Class II antigens in mediating eosinophil functions.


Asunto(s)
Proteínas de Unión al ADN , Eosinófilos/metabolismo , Regulación Neoplásica de la Expresión Génica , Regulación de la Expresión Génica , Glucocorticoides/farmacología , Proteínas Represoras , Ribonucleasas , Factores de Transcripción , Regulación hacia Arriba , Adulto , Apoptosis , Araquidonato 15-Lipooxigenasa/biosíntesis , Proteínas Sanguíneas/biosíntesis , Northern Blotting , Western Blotting , Diferenciación Celular , Separación Celular , ADN Complementario/metabolismo , Dexametasona/farmacología , Relación Dosis-Respuesta a Droga , Proteínas en los Gránulos del Eosinófilo , Citometría de Flujo , Genes MHC Clase II , Humanos , Etiquetado Corte-Fin in Situ , Interleucina-5/antagonistas & inhibidores , Masculino , Análisis de Secuencia por Matrices de Oligonucleótidos , Prostaglandina-Endoperóxido Sintasas/biosíntesis , Proteína Proto-Oncogénica c-ets-2 , Proteínas Proto-Oncogénicas/biosíntesis , ARN/metabolismo , Transducción de Señal , Factores de Tiempo , Transactivadores/biosíntesis , Células Tumorales Cultivadas
16.
J Allergy Clin Immunol ; 111(1): 3-22; quiz 23, 2003 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-12532089

RESUMEN

Glucocorticoids (GCs) are the most common group of medications used in the treatment of allergic and autoimmune disorders. They produce potent anti-inflammatory effects by inducing or repressing the expression of target genes. Although most patients with allergic diseases and autoimmune disorders respond to GC therapy, a small subset of patients demonstrate persistent tissue inflammation despite treatment with high doses of GCs. This condition results from an interaction between susceptibility genes, the host's environment, and immunologic factors. The treatment of these patients requires a systematic approach to rule out underlying conditions that lead to steroid resistance or treatment failure, as well as the use of alternative strategies to inhibit tissue inflammation.


Asunto(s)
Glucocorticoides/farmacología , Glucocorticoides/uso terapéutico , Enfermedades Autoinmunes/tratamiento farmacológico , Resistencia a Medicamentos/genética , Hipersensibilidad/tratamiento farmacológico
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