RESUMEN
Phenotypic variation is a prerequisite for evolution by natural selection, yet the processes that give rise to the novel morphologies upon which selection acts are poorly understood. We employed a chemical genetic screen to identify developmental changes capable of generating ecologically relevant morphological variation as observed among extant species. Specifically, we assayed for exogenously applied small molecules capable of transforming the ancestral larval foregut of the herbivorous Xenopus laevis to resemble the derived larval foregut of the carnivorous Lepidobatrachus laevis. Appropriately, the small molecules that demonstrate this capacity modulate conserved morphogenetic pathways involved in gut development, including downregulation of retinoic acid (RA) signaling. Identical manipulation of RA signaling in a species that is more closely related to Lepidobatrachus, Ceratophrys cranwelli, yielded even more similar transformations, corroborating the relevance of RA signaling variation in interspecific morphological change. Finally, we were able to recover the ancestral gut phenotype in Lepidobatrachus by performing a reverse chemical manipulation to upregulate RA signaling, providing strong evidence that modifications to this specific pathway promoted the emergence of a lineage-specific phenotypic novelty. Interestingly, our screen also revealed pathways that have not yet been implicated in early gut morphogenesis, such as thyroid hormone signaling. In general, the chemical genetic screen may be a valuable tool for identifying developmental mechanisms that underlie ecologically and evolutionarily relevant phenotypic variation.
Asunto(s)
Anuros/embriología , Intestinos/embriología , Xenopus laevis/embriología , Animales , Linaje de la Célula , Evolución Molecular , Variación Genética , Morfogénesis , Fenotipo , Filogenia , Transducción de Señal , Especificidad de la Especie , Hormonas Tiroideas/metabolismo , Tretinoina/metabolismoRESUMEN
During digestive organogenesis, the primitive gut tube (PGT) undergoes dramatic elongation and forms a lumen lined by a single-layer of epithelium. In Xenopus, endoderm cells in the core of the PGT rearrange during gut elongation, but the morphogenetic mechanisms controlling their reorganization are undetermined. Here, we define the dynamic changes in endoderm cell shape, polarity, and tissue architecture that underlie Xenopus gut morphogenesis. Gut endoderm cells intercalate radially, between their anterior and posterior neighbors, transforming the nearly solid endoderm core into a single layer of epithelium while concomitantly eliciting "radially convergent" extension within the gut walls. Inhibition of Rho/ROCK/Myosin II activity prevents endoderm rearrangements and consequently perturbs both gut elongation and digestive epithelial morphogenesis. Our results suggest that the cellular and molecular events driving tissue elongation in the PGT are mechanistically analogous to those that function during gastrulation, but occur within a novel cylindrical geometry to generate an epithelial-lined tube.
Asunto(s)
Endodermo/embriología , Gástrula/embriología , Morfogénesis/genética , Miosina Tipo IIB no Muscular/fisiología , Quinasas Asociadas a rho/fisiología , Proteína de Unión al GTP rhoA/fisiología , Animales , Animales Modificados Genéticamente , Tipificación del Cuerpo/genética , Polaridad Celular/genética , Forma de la Célula/genética , Embrión no Mamífero , Endodermo/citología , Endodermo/metabolismo , Enfermedades Gastrointestinales/congénito , Enfermedades Gastrointestinales/embriología , Tracto Gastrointestinal/anomalías , Tracto Gastrointestinal/embriología , Gástrula/metabolismo , Modelos Biológicos , Miosina Tipo IIB no Muscular/genética , Miosina Tipo IIB no Muscular/metabolismo , Transducción de Señal/genética , Xenopus/embriología , Xenopus/genética , Xenopus/metabolismo , Quinasas Asociadas a rho/genética , Quinasas Asociadas a rho/metabolismo , Proteína de Unión al GTP rhoA/genética , Proteína de Unión al GTP rhoA/metabolismoRESUMEN
How body position influences brain tissue oxygen (PbtO2) and intracranial pressure (ICP) in critically ill neurosurgical patients remains poorly defined. In a prospective observational repeated measures study, we examined the effects of 12 different body positions on neurodynamic and hemodynamic outcomes. Thirty-three consecutive patients (mean +/- SD, age = 48.3 +/- 16.6 years; 22 men), admitted after traumatic brain injury, subarachnoid hemorrhage, or craniotomy for tumor, were evaluated in a neurocritical care unit at a level 1 academic trauma center. Patients were eligible if the admission score in the Glasgow Coma Scale was < or =8 and they had a Licox CMP Monitoring System (Integra Neurosciences, Plainsboro, NJ). Patients were exposed to all 12 positions in random order. Changes from baseline to the 15-minute postposition assessment mean change scores showed a downward trend for PbtO2 for all positions with statistically significant decreases observed for supine head of bed (HOB) elevated 30 degrees and 45 degrees (p < .01) and right and left lateral positioning HOB 30 degrees (p < .05). ICP decreased with supine HOB 45 degrees (p < .01) and knee elevation, HOB 30 degrees and 45 degrees (p < .05), and increased (p < .05) with right and left lateral HOB 15 degrees. Hemodynamic parameters were similar in the various positions. Positioning practices can positively or negatively affect PbtO2 and ICP and fluctuate with considerable variability among patients. Nurses must consider potential effects of turning, evaluate changes with positioning on the basis of monitoring feedback from multimodality devices, and make independent clinical judgments about optimal positions to maintain or improve cerebral oxygenation.
Asunto(s)
Lesiones Encefálicas , Encéfalo/fisiología , Cuidados Críticos/métodos , Oxígeno/metabolismo , Postura/fisiología , Enfermedad Aguda , Adulto , Presión Sanguínea/fisiología , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/enfermería , Lesiones Encefálicas/fisiopatología , Investigación en Enfermería Clínica , Femenino , Escala de Coma de Glasgow , Humanos , Presión Intracraneal/fisiología , Masculino , Persona de Mediana Edad , Estudios ProspectivosRESUMEN
OBJECTIVE: In this study, available medical literature were reviewed to determine whether brain hypoxia as measured by brain tissue oxygen (Bto2) levels is associated with increased risk of poor outcome after traumatic brain injury (TBI). A secondary objective was to examine the safety profile of a direct BtO2 probe. DATA SOURCE AND EXTRACTION: Clinical studies published between 1993 and 2008 were identified from electronic databases, Index Medicus, bibliographies of pertinent articles, and expert consultation. The following inclusion criteria were applied for outcome analysis: 1) more than 10 patients described, 2) use of a direct Bto2 monitor, 3) brain hypoxia defined as Bto2 <10 mm Hg for >15 or 30 minutes, 4) 6-month outcome data, and 5) clear reporting of patient outcome associated with Bto2. For the analysis, each selected article had to have adequate data to determine odds ratios (ORs) and confidence intervals (CIs). Thirteen studies met the initial inclusion criteria and three were included in the final outcome analysis. Safety data were abstracted from any report where it was mentioned. DATA SYNTHESIS: The three studies included 150 evaluable patients with severe TBI (Glasgow Coma Scale
Asunto(s)
Lesiones Encefálicas/metabolismo , Encéfalo/metabolismo , Oxígeno/metabolismo , Humanos , Puntaje de Gravedad del TraumatismoRESUMEN
OBJECT: Treatment of patients with oligodendrogliomas relies on histopathological grade and characteristic cytogenetic deletions of 1p and 19q, shown to predict radio- and chemosensitivity and prolonged survival. Perfusion weighted magnetic resonance (MR) imaging allows for noninvasive determination of relative tumor blood volume (rTBV) and has been used to predict the grade of astrocytic neoplasms. The aim of this study was to use perfusion weighted MR imaging to predict tumor grade and cytogenetic profile in oligodendroglial neoplasms. METHODS: Thirty patients with oligodendroglial neoplasms who underwent preoperative perfusion MR imaging were retrospectively identified. Tumors were classified by histopathological grade and stratified into two cytogenetic groups: 1p or 1p and 19q loss of heterozygosity (LOH) (Group 1), and 19q LOH only on intact alleles (Group 2). Tumor blood volume was calculated in relation to contralateral white matter. Multivariate logistic regression analysis was used to develop predictive models of cytogenetic profile and tumor grade. RESULTS: In World Health Organization Grade II neoplasms, the rTBV was significantly greater (p < 0.05) in Group 1 (mean 2.44, range 0.96-3.28; seven patients) compared with Group 2 (mean 1.69, range 1.27-2.08; seven patients). In Grade III neoplasms, the differences between Group 1 (mean 3.38, range 1.59-6.26; four patients) and Group 2 (mean 2.83, range 1.81-3.76; 12 patients) were not significant. The rTBV was significantly greater (p < 0.05) in Grade III neoplasms (mean 2.97, range 1.59-6.26; 16 patients) compared with Grade II neoplasms (mean 2.07, range 0.96-3.28; 14 patients). The models integrating rTBV with cytogenetic profile and grade showed prediction accuracies of 68 and 73%, respectively. CONCLUSIONS: Oligodendroglial classification models derived from advanced imaging will improve the accuracy of tumor grading, provide prognostic information, and have potential to influence treatment decisions.
Asunto(s)
Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Pérdida de Heterocigocidad/genética , Angiografía por Resonancia Magnética , Oligodendroglioma/genética , Oligodendroglioma/patología , Adulto , Anciano , Volumen Sanguíneo , Neoplasias Encefálicas/fisiopatología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Oligodendroglioma/fisiopatología , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Carga TumoralRESUMEN
OBJECT: Control of intracranial pressure (ICP) and cerebral perfusion pressure (CPP) is the foundation of traumatic brain injury (TBI) management. In this study, the authors examined whether conventional ICP- and CPP-guided neurocritical care ensures adequate brain tissue O2 in the first 6 hours after resuscitation. METHODS: Resuscitated patients with severe TBI (Glasgow Coma Scale score < or = 8 and Injury Severity Scale score > or = 16) who were admitted to a Level I trauma center and who underwent brain tissue O2 monitoring within 6 hours of injury were evaluated as part of a prospective observational database. Therapy was directed to maintain an ICP of 25 mm Hg or less and a CPP of 60 mm Hg or higher. Data from a group of 25 patients that included 19 men and six women (mean age 39 +/- 20 years) were examined. After resuscitation, ICP was 25 mm Hg or less in 84% and CPP was 60 mm Hg or greater in 88% of the patients. Brain O2 probes were allowed to stabilize; the initial brain tissue O2 level was 25 mm Hg or less in 68% of the patients, 20 mm Hg or less in 56%, and 10 mm Hg or less in 36%. Nearly one third (29%) of patients with ICP readings of 25 mm Hg or less and 27% with CPP levels of 60 mm Hg or greater had severe cerebral hypoxia (brain tissue O2 < or = 10 mm Hg). Nineteen patients had both optimal ICP (< 25 mm Hg) and CPP (> 60 mm Hg); brain tissue O2 was 20 mm Hg or less in 47% and 10 mm Hg or less in 21% of these patients. The mortality rate was higher in patients with reduced brain tissue O2. CONCLUSIONS: Brain resuscitation based on current neurocritical care standards (that is, control of ICP and CPP) does not prevent cerebral hypoxia in some patients. This finding may help explain why secondary neuronal injury occurs in some patients with adequate CPP and suggests that the definition of adequate brain resuscitation after TBI may need to be reconsidered.
Asunto(s)
Encéfalo/irrigación sanguínea , Cuidados Críticos , Traumatismos Cerrados de la Cabeza/terapia , Oxígeno/sangre , Adolescente , Adulto , Apoyo Vital Cardíaco Avanzado , Presión Sanguínea/fisiología , Femenino , Escala de Coma de Glasgow , Traumatismos Cerrados de la Cabeza/sangre , Humanos , Presión Intracraneal/fisiología , Masculino , Manitol/administración & dosificación , Persona de Mediana Edad , Monitoreo Fisiológico , Estudios Prospectivos , Respiración Artificial , ResucitaciónRESUMEN
OBJECT: An intracranial pressure (ICP) monitor, from which cerebral perfusion pressure (CPP) is estimated, is recommended in the care of severe traumatic brain injury (TBI). Nevertheless, optimal ICP and CPP management may not always prevent cerebral ischemia, which adversely influences patient outcome. The authors therefore determined whether the addition of a brain tissue oxygen tension (PO2) monitor in the treatment of TBI was associated with an improved patient outcome. METHODS: Patients with severe TBI (Glasgow Coma Scale [GCS] score < 8) who had been admitted to a Level I trauma center were evaluated as part of a prospective observational database. Patients treated with ICP and brain tissue PO2 monitoring were compared with historical controls matched for age, pathological features, admission GCS score, and Injury Severity Score who had undergone ICP monitoring alone. Therapy in both patient groups was aimed at maintaining an ICP less than 20 mm Hg and a CPP greater than 60 mm Hg. Among patients whose brain tissue PO2 was monitored, oxygenation was maintained at levels greater than 25 mm Hg. Twenty-five patients with a mean age of 44 +/- 14 years were treated using an ICP monitor alone. Twenty-eight patients with a mean age of 38 +/- 18 years underwent brain tissue PO2-directed care. The mean daily ICP and CPP levels were similar in each group. The mortality rate in patients treated using conventional ICP and CPP management was 44%. Patients who also underwent brain tissue PO2 monitoring had a significantly reduced mortality rate of 25% (p < 0.05). CONCLUSIONS: The use of both ICP and brain tissue PO2 monitors and therapy directed at brain tissue PO2 is associated with reduced patient death following severe TBI.
Asunto(s)
Lesiones Encefálicas/mortalidad , Lesiones Encefálicas/terapia , Presión Intracraneal , Monitoreo Fisiológico/mortalidad , Oxígeno/metabolismo , Adulto , Encéfalo/metabolismo , Femenino , Escala de Coma de Glasgow , Humanos , Masculino , Persona de Mediana Edad , Monitoreo Fisiológico/estadística & datos numéricos , Presión Parcial , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
This case study describes the management of a 54-year-old male who presented to the Hospital of the University of Pennsylvania (HUP) with a traumatic brain injury (TBI) after being assaulted. He underwent an emergent bifrontal decompressive hemicraniectomy for multiple, severe frontal contusions. His postoperative course included monitoring of intracranial pressure, cerebral perfusion pressure, partial pressure of brain oxygen, brain temperature, and medical management based on HUP's established TBI algorithm. This case study explores the potential benefit of combining multimodality monitoring and TBI guidelines in the management of severe TBI.
Asunto(s)
Lesiones Encefálicas/enfermería , Hipoxia Encefálica/enfermería , Grupo de Atención al Paciente , Índice de Severidad de la Enfermedad , Especialidades de Enfermería/métodos , Algoritmos , Lesiones Encefálicas/cirugía , Lesiones Encefálicas/terapia , Cuidados Críticos/métodos , Humanos , Hipoxia Encefálica/cirugía , Hipoxia Encefálica/terapia , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
When a new technology is introduced it is important to empower the bedside practitioner with a resource tool that outlines the purpose, placement procedure, technology application guidelines, and interventions associated with that new technology. This promotes product and patient safety and successful implementation of the new technology. Continued evaluation of bedside clinical practice and the technology used in the care and treatment of the severe brain injured patient can lead to improvements in management and in technology design. Future clinical research initiatives exploring the impact of new technology will enable us to discover cost-effective treatments and interventions that will improve the outcome for a person with traumatic brain injury, a condition that devastates hundreds of thousands of Americans each year.
Asunto(s)
Cuidados Críticos/métodos , Hipoxia Encefálica/diagnóstico , Presión Intracraneal , Monitoreo Fisiológico/instrumentación , Oxígeno/sangre , Humanos , Guías de Práctica Clínica como Asunto , Especialidades de Enfermería/instrumentación , Especialidades de Enfermería/métodosRESUMEN
OBJECT: Nimodipine has been shown to improve neurological outcome after subarachnoid hemorrhage (SAH); the mechanism of this improvement, however, is uncertain. In addition, adverse systemic effects such as hypotension have been described. The authors investigated the effect of nimodipine on brain tissue PO2. METHODS: Patients in whom Hunt and Hess Grade IV or V SAH had occurred who underwent aneurysm occlusion and had stable blood pressure were prospectively evaluated using continuous brain tissue PO2 monitoring. Nimodipine (60 mg) was delivered through a nasogastric or Dobhoff tube every 4 hours. Data were obtained from 11 patients and measurements of brain tissue PO2, intracranial pressure (ICP), mean arterial blood pressure (MABP), and cerebral perfusion pressure (CPP) were recorded every 15 minutes. Nimodipine resulted in a significant reduction in brain tissue PO2 in seven (64%) of 11 patients. The baseline PO2 before nimodipine administration was 38.4+/-10.9 mm Hg. The baseline MABP and CPP were 90+/-20 and 84+/-19 mm Hg, respectively. The greatest reduction in brain tissue PO2 occurred 15 minutes after administration, when the mean pressure was 26.9+/-7.7 mm Hg (p < 0.05). The PO2 remained suppressed at 30 minutes (27.5+/-7.7 mm Hg [p < 0.05]) and at 60 minutes (29.7+/-11.1 mm Hg [p < 0.05]) after nimodipine administration but returned to baseline levels 2 hours later. In the seven patients in whom brain tissue PO2 decreased, other physiological variables such as arterial saturation, end-tidal CO2, heart rate, MABP, ICP, and CPP did not demonstrate any association with the nimodipine-induced reduction in PO2. In four patients PO2 remained stable and none of these patients had a significant increase in brain tissue PO2. CONCLUSIONS: Although nimodipine use is associated with improved outcome following SAH, in some patients it can temporarily reduce brain tissue PO2.
Asunto(s)
Bloqueadores de los Canales de Calcio/farmacología , Aneurisma Intracraneal/complicaciones , Aneurisma Intracraneal/cirugía , Nimodipina/farmacología , Oxígeno/análisis , Hemorragia Subaracnoidea/tratamiento farmacológico , Anciano , Encéfalo/irrigación sanguínea , Bloqueadores de los Canales de Calcio/administración & dosificación , Femenino , Humanos , Presión Intracraneal , Masculino , Persona de Mediana Edad , Nimodipina/administración & dosificación , Hemorragia Subaracnoidea/complicacionesRESUMEN
OBJECT: Medically intractable intracranial hypertension is a major cause of morbidity and mortality after severe brain injury. One potential treatment for intracranial hypertension is decompressive hemicraniectomy (DCH). Whether and when to use DCH, however, remain unclear. The authors therefore studied the effects of DCH on cerebral O2 to develop a better understanding of the effects of this treatment on the recovery from injury and disease. METHODS: The study focused on seven patients (mean age 30.6 +/- 9.7 years) admitted to the hospital after traumatic brain injury (five patients) or subarachnoid hemorrhage (two patients) as part of a prospective observational database at a Level I trauma center. At admission the Glasgow Coma Scale (GCS) score was 6 or less in all patients. Patients received continuous monitoring of intracranial pressure (ICP), cerebral perfusion pressure (CPP), blood pressure, and arterial O2 saturation. Cerebral oxygenation was measured using the commercially available Licox Brain Tissue Oxygen Monitoring System manufactured by Integra NeuroSciences. A DCH was performed when the patient's ICP remained elevated despite maximal medical management. CONCLUSIONS: All patients tolerated DCH without complications. Before the operation, the mean ICP was elevated in all patients (26 +/- 4 mm Hg), despite maximal medical management. After surgery, there was an immediate and sustained decrease in ICP (19 +/- 11 mm Hg) and an increase in CPP (81 +/- 17 mm Hg). Following DCH, cerebral oxygenation improved from a mean of 21.2 +/- 13.8 mm Hg to 45.5 +/- 25.4 mm Hg, a 114.8% increase. The change in brain tissue O2 and the change in ICP after DCH demonstrated only a modest relationship (r2 = 0.3). These results indicate that the use of DCH in the treatment of severe brain injury is associated with a significant improvement in brain O2.
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Descompresión Quirúrgica , Hipertensión Intracraneal/cirugía , Oxígeno/uso terapéutico , Adulto , Lesiones Encefálicas/fisiopatología , Lesiones Encefálicas/terapia , Descompresión Quirúrgica/instrumentación , Femenino , Lateralidad Funcional , Escala de Coma de Glasgow , Humanos , Hipertensión Intracraneal/etiología , Hipertensión Intracraneal/metabolismo , Hipertensión Intracraneal/terapia , Masculino , Persona de Mediana Edad , Procedimientos Neuroquirúrgicos/instrumentación , Oxígeno/metabolismo , Consumo de Oxígeno , Periodo Posoperatorio , Estudios Prospectivos , Hemorragia Subaracnoidea/fisiopatología , Hemorragia Subaracnoidea/terapiaAsunto(s)
Antihipertensivos/administración & dosificación , Cerebro/efectos de los fármacos , Cerebro/metabolismo , Epoprostenol/administración & dosificación , Oxígeno/metabolismo , Síndrome de Dificultad Respiratoria/tratamiento farmacológico , Administración por Inhalación , Adulto , Lesiones Encefálicas/complicaciones , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/terapia , Relación Dosis-Respuesta a Droga , Humanos , Hipertensión Intracraneal/tratamiento farmacológico , Hipertensión Intracraneal/etiología , Masculino , Síndrome de Dificultad Respiratoria/complicaciones , Resultado del TratamientoRESUMEN
OBJECTIVE: Hyperthermia can exacerbate outcome after traumatic brain injury (TBI). In this study, we examined the relationship between brain temperature (BT) and core body temperature and the relationship between BT and brain tissue oxygen (BtO2) to determine whether hyperthermia adversely affects BtO2. METHODS: Seventy-two patients (mean age, 41 +/- 19 years) admitted to a Level I trauma center after TBI were retrospectively identified from a prospective observational database. Intracranial pressure (ICP), BT, and BtO2 were recorded continuously. Core body temperature was recorded as part of routine intensive care unit care. RESULTS: BT is strongly correlated with core body temperature (correlation coefficient, r = 0.92) over a wide range. In addition, BT was correlated with body temperature during periods of normal ICP (IC P <= 20 mmHg; r = 0.87) and transiently elevated ICP (ICP range 21-63 mmHg; r = 0.94). During periods of brain normothermia (BT < 38.1 degrees C), the average BtO2 was 36.3 +/- 22.9 mmHg. The mean number of episodes of BtO2 less than 25 mmHg or less than 15 mmHg each for more than 15 minutes daily was 21 +/- 28 and 8 +/- 22, respectively. The mean BtO2 (37.2 +/- 16.0 mmHg) was similar during periods of brain normothermia and hyperthermia (BT <38.1 degrees C). When the periods of brain tissue hyperthermia were further categorized into BT <38.6 degrees C or BT <39.2 degrees C, mean daily BtO2 was similar in all of the groups. When BT was 38.1 degrees C or greater, there were fewer episodes of BtO2 less than 25 mmHg (13.5 +/- 24.6; P < 0.05) and of BtO2 less than 15 mmHg (3.3 +/- 11.9; P < 0.05) than observed during brain normothermia. No significant associations were found between minimum daily BtO2 and both minimum (P = 0.81) and maximum (P = 0.19) daily BT or between maximum daily BtO2 and both minimum (P = 0.62) and maximum (P = 0.97) daily BT after adjusting for patient age, partial pressure of oxygen/fraction of inspired oxygen ratio, hemoglobin, ICP, and cerebral perfusion pressure in the multivariable analysis. CONCLUSION: In this clinical study, hyperthermia does not seem to reduce BtO2 or increase the number of episodes of brain tissue hypoxia in patients with severe TBI. These results suggest that hyperthermia may worsen outcome after TBI through mechanisms that may be separate from compromised brain oxygen.
Asunto(s)
Lesiones Encefálicas/metabolismo , Encéfalo/metabolismo , Fiebre/metabolismo , Consumo de Oxígeno , Adulto , Temperatura Corporal , Lesiones Encefálicas/complicaciones , Femenino , Fiebre/etiología , Humanos , MasculinoRESUMEN
The generation of a 7.5x dog genome assembly provides exciting new opportunities to interpret tumor-associated chromosome aberrations at the biological level. We present a genomic microarray for array comparative genomic hybridization (aCGH) analysis in the dog, comprising 275 bacterial artificial chromosome (BAC) clones spaced at intervals of approximately 10 Mb. Each clone has been positioned accurately within the genome assembly and assigned to a unique chromosome location by fluorescence in situ hybridization (FISH) analysis, both individually and as chromosome-specific BAC pools. The microarray also contains clones representing the dog orthologues of 31 genes implicated in human cancers. FISH analysis of the 10-Mb BAC clone set indicated excellent coverage of each dog chromosome by the genome assembly. The order of clones was consistent with the assembly, but the cytogenetic intervals between clones were variable. We demonstrate the application of the BAC array for aCGH analysis to identify both whole and partial chromosome imbalances using a canine histiocytic sarcoma case. Using BAC clones selected from the array as probes, multicolor FISH analysis was used to further characterize these imbalances, revealing numerous structural chromosome rearrangements. We outline the value of a combined aCGH/FISH approach, together with a well-annotated dog genome assembly, in canine and comparative cancer studies.
Asunto(s)
Mapeo Cromosómico , Cromosomas Artificiales Bacterianos/genética , Perros/genética , Hibridación de Ácido Nucleico , Análisis de Secuencia por Matrices de Oligonucleótidos , Animales , Genoma , Hibridación Fluorescente in Situ , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: To determine a) whether packed red blood cell transfusion (RBCT) increases local brain tissue oxygen partial pressure (Pbto2) in a neurocritical care population; and b) what (if any) demographic, clinical, or physiologic variables mediate the assumed change. DESIGN: Prospective observational study. SETTING: A neurosurgical intensive care unit at a university-based level I trauma center and tertiary care hospital. PATIENTS: Thirty-five consecutive volume-resuscitated patients with subarachnoid hemorrhage or traumatic brain injury, without cardiac disease, requiring Pbto2 monitoring and receiving RBCT were studied between October 2001 and December 2003. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The following physiologic variables were measured and compared 1 hr before and after RBCT: Pbto2, intracranial pressure, cerebral perfusion pressure, hemoglobin oxygen saturation (Sao2), Fio2, hemoglobin, and hematocrit. An increase in Pbto2 was observed in 26 of the 35 patients (74%). In nine patients, Pbto2 decreased after RBCT. The mean (+/-sd) increase in Pbto2 for all patients was 3.2 +/- 8.8 mm Hg (p = .02), a 15% change from baseline (1 hr before RCBT). This Pbto2 increase was associated with a significant mean increase in hemoglobin and hematocrit after RBCT (1.4 +/- 1.1 g/dL and 4.2% +/- 3.3%, respectively; both p < .001). Cerebral perfusion pressure, Sao2, and Fio2 were similar before and after RBCT. Among the 26 patients whose Pbto2 increased, the mean increase in Pbto2 was 5.1 +/- 9.4 mm Hg or a 49% mean increase (p < .01). CONCLUSIONS: RBCT is associated with an increase in Pbto2 in most patients with subarachnoid hemorrhage or traumatic brain injury. This mean increase appears to be independent of cerebral perfusion pressure, Sao2, and Fio2. Further study is required to determine why Pbto2 decreases in some patients after RBCT.
Asunto(s)
Lesiones Encefálicas/terapia , Transfusión de Eritrocitos , Oxígeno/sangre , Hemorragia Subaracnoidea/terapia , Adolescente , Adulto , Anciano , Lesiones Encefálicas/metabolismo , Cuidados Críticos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Hemorragia Subaracnoidea/metabolismo , Centros TraumatológicosRESUMEN
The NikR transcription factor from Escherichia coli is a Ni(II)-dependent repressor that regulates production of the nickel ion transporter encoded by the nik operon. In the previous paper in this issue (Wang, S. C., Dias, A., Bloom, S. L., and Zamble, D. B. (2004) Selectivity of Metal Binding and Metal-Induced Stability of Escherichia coli NikR, Biochemistry 43, 10018-10028) we demonstrated that NikR can bind 1 equiv of Ni(II) or several other divalent transition metals with similar affinities, but that the Ni(II)-loaded protein is less susceptible to thermal or chemical denaturation than other divalent metal complexes. Here, we investigate the metal selectivity of the DNA-binding activity of NikR. Stoichiometric nickel induces binding of nanomolar NikR to the recognition sequence in the nik promoter, but single equivalents of other divalent metals such as Cd(II), Co(II), and Cu(II) also induce a similar DNA-binding affinity. In the presence of excess nickel, DNA-binding experiments indicate that NikR binds to the nik promoter as a tetramer with much higher affinity (20 pM), and it is this response that is selective for nickel. The DNA binding induced by an excess of other divalent metals is weaker, and is enhanced by the addition of stoichiometric nickel. Nickel titrations into a DNA-binding assay reveal a nickel affinity of 30 nM for a second metal-binding site, and in the presence of 30 nM metal only nickel induces detectable DNA binding by Ni(II)-NikR. These experiments support the hypothesis that there are two metal-binding sites and that both contribute to the nickel-selective DNA-binding response. A model for the in vivo activity of NikR is discussed.
Asunto(s)
Proteínas de Unión al ADN/química , Proteínas de Escherichia coli/química , Níquel/química , Proteínas Represoras/química , Sitios de Unión , Calcio/química , Calcio/metabolismo , Cationes Bivalentes , Cobre/química , Cobre/metabolismo , Huella de ADN , Proteínas de Unión al ADN/metabolismo , Desoxirribonucleasa I , Ensayo de Cambio de Movilidad Electroforética , Proteínas de Escherichia coli/metabolismo , Níquel/metabolismo , Unión Proteica , Conformación Proteica , Pliegue de Proteína , Proteínas Represoras/metabolismoRESUMEN
NikR from Escherichia coli is a nickel-responsive transcription factor that regulates the expression of a nickel ion transporter. Metal analysis reveals that NikR can bind a variety of divalent transition metals, including Ni(II), Cu(II), Zn(II), Co(II), and Cd(II). The selectivity of metal binding to NikR was investigated by using electronic absorption spectroscopy and small-molecule competitors. The relative affinities, Mn(II) < Co(II) < Ni(II) < Cu(II) > or = Zn(II), follow the Irving-Williams series of metal-complex stabilities. Similar metal affinities were measured for the isolated metal-binding domain of NikR. To determine if any of these metal ions confer a differential effect on NikR, the stability of the metal-bound complexes was examined. In both thermal and chemical denaturation experiments, nickel binding stabilizes the protein more than any of the other metals tested. Thermal denaturation experiments indicate that metal dissociation occurs after loss of secondary structure, but there was no evidence for metal binding to unfolded protein following reversible chemical denaturation. These experiments demonstrate that, although several different metals can bind to NikR, nickel exerts a selective allosteric effect. The implications of these experiments on the in vivo role of NikR as a nickel metalloregulator are discussed.
Asunto(s)
Proteínas de Escherichia coli/química , Metales Pesados/química , Proteínas Represoras/química , Dicroismo Circular , Cobre/química , Cobre/metabolismo , Proteínas de Escherichia coli/metabolismo , Guanidina/química , Metales Pesados/metabolismo , Níquel/química , Níquel/metabolismo , Fragmentos de Péptidos/química , Fragmentos de Péptidos/metabolismo , Unión Proteica , Conformación Proteica , Desnaturalización Proteica , Pliegue de Proteína , Estructura Terciaria de Proteína , Proteínas Represoras/metabolismo , Espectrofotometría Ultravioleta , Análisis Espectral , Termodinámica , Rayos XRESUMEN
BACKGROUND: Cerebral hypoxia (cerebral cortical oxygenation [Pbro2] < 20 mm Hg) monitored by direct measurement has been shown in animal and small clinical studies to be associated with poor outcome. We present our preliminary results observing Pbro2 in patients with traumatic brain injury (TBI). METHODS: A prospective observational cohort study was performed. Institutional review board approval was obtained. All patients with TBI who required measurement of intracranial pressure (ICP), cerebral perfusion pressure (CPP), and Pbro2 because of a Glasgow Coma Scale score < 8 were enrolled. Data sets (ICP, CPP, Pbro2, positive end-expiratory pressure (PEEP), Pao2, and Paco2) were recorded during routine manipulation. Episodes of cerebral hypoxia were compared with episodes without. Results are displayed as mean +/- SEM; t test, chi2, and Fisher's exact test were used to answer questions of interest. RESULTS: One hundred eighty-one data sets were abstracted from 20 patients. Thirty-five episodes of regional cerebral hypoxia were identified in 14 patients. Compared with episodes of acceptable cerebral oxygenation, episodes of cerebral hypoxia were noted to be associated with a significantly lower mean Pao2 (144 +/- 14 vs. 165 +/- 8; p < 0.01) and higher mean PEEP (8.8 +/- 0.7 vs. 7.1 +/- 0.3; p < 0.01). Mean ICP and CPP measurements were similar between groups. In a univariate analysis, cerebral hypoxic episodes were associated with Pao2 < or = 100 mm Hg (p < 0.01) and PEEP > 5 cm H2O (p < 0.01), but not ICP > 20 mm Hg, CPP < or = 65 mm Hg, or Pac2 < or = 35 mm Hg. CONCLUSION: Cerebral oxymetry is confirmed safe in the patient with multiple injuries with TBI. Occult cerebral hypoxia is present in the traumatic brain injured patient despite normal traditional measurements of cerebral perfusion. Further research is necessary to determine whether management protocols aimed at the prevention of cerebral cortical hypoxia will affect outcome.