RESUMO
Rett syndrome (RTT) is a debilitating neurological disorder caused by mutations in the gene encoding the transcription factor Methyl CpG Binding Protein 2 (MECP2). A distinct disorder results from MECP2 gene duplication, suggesting that therapeutic approaches must restore close to normal levels of MECP2. Here, we apply the approach of site-directed RNA editing to repair, at the mRNA level, a disease-causing guanosine to adenosine (G > A) mutation in the mouse MeCP2 DNA binding domain. To mediate repair, we exploit the catalytic domain of Adenosine Deaminase Acting on RNA (ADAR2) that deaminates A to inosine (I) residues that are subsequently translated as G. We fuse the ADAR2 domain, tagged with a nuclear localization signal, to an RNA binding peptide from bacteriophage lambda. In cultured neurons from mice that harbor an RTT patient G > A mutation and express engineered ADAR2, along with an appropriate RNA guide to target the enzyme, 72% of Mecp2 mRNA is repaired. Levels of MeCP2 protein are also increased significantly. Importantly, as in wild-type neurons, the repaired MeCP2 protein is enriched in heterochromatic foci, reflecting restoration of normal MeCP2 binding to methylated DNA. This successful use of site-directed RNA editing to repair an endogenous mRNA and restore protein function opens the door to future in vivo applications to treat RTT and other diseases.
Assuntos
Proteína 2 de Ligação a Metil-CpG/genética , Neurônios/fisiologia , RNA/genética , Adenosina Desaminase/genética , Animais , Células Cultivadas , Metilação de DNA/genética , Modelos Animais de Doenças , Humanos , Camundongos , Mutação/genética , RNA Mensageiro/genética , Proteínas de Ligação a RNA/genética , Síndrome de Rett/genéticaRESUMO
Programmable RNA editing is gaining momentum as an approach to repair mutations, but its efficiency in repairing endogenous mutant RNA in complex tissue is unknown. Here we apply this approach to the brain and successfully repair a guanosine-to-adenosine mutation in methyl CpG binding protein 2 RNA that causes the neurodevelopmental disease Rett syndrome. Repair is mediated by hippocampal injections of juvenile Mecp2317G>A mice with an adeno-associated virus expressing the hyperactive catalytic domain of adenosine deaminase acting on RNA 2 and Mecp2 guide. After 1 month, 50% of Mecp2 RNA is recoded in three different hippocampal neuronal populations. MeCP2 protein localization to heterochromatin is restored in neurons to 50% of wild-type levels. Whole-transcriptome RNA analysis of one neuronal population indicates that the majority of off-target editing sites exhibit rates of 30% or less. This study demonstrates that programmable RNA editing can be utilized to repair mutations in mouse models of neurological disease.
Assuntos
Terapia Genética , Proteína 2 de Ligação a Metil-CpG/genética , Doenças do Sistema Nervoso/genética , Doenças do Sistema Nervoso/terapia , Edição de RNA/genética , Sequência de Aminoácidos , Animais , Linhagem Celular , Perfilação da Expressão Gênica , Células HEK293 , Heterocromatina/metabolismo , Hipocampo/metabolismo , Humanos , Masculino , Proteína 2 de Ligação a Metil-CpG/química , Camundongos , RNA/genética , Técnicas EstereotáxicasRESUMO
Aim: To compare perioperative opioid consumption for patients undergoing mastectomy surgery with or without pectoralis nerve (PECS) plane blocks. Patients & methods: Retrospective study evaluating 152 adult females with mastectomies. Demographics, postanesthesia care unit stay duration and opioid consumption data at three time points were collected and analyzed for statistical significance. Results: 98 patients were included in the PECS block group, 54 patients were in the general anesthesia only group. Age and BMI were comparable. Total perioperative intravenous opioid consumption was less in the PECS block group (50.88 mg) compared with the general anesthesia only group (67.83 mg), p < 0.001. Conclusion: Acute pain after mastectomy is often severe. PECS plane block may decrease perioperative opioid consumption after mastectomy surgery compared with general anesthesia alone.
Assuntos
Dor Aguda , Analgésicos Opioides/administração & dosagem , Anestesia Geral/estatística & dados numéricos , Neoplasias da Mama/cirurgia , Mastectomia/estatística & dados numéricos , Bloqueio Nervoso/estatística & dados numéricos , Manejo da Dor/estatística & dados numéricos , Dor Pós-Operatória , Cuidados Pré-Operatórios/estatística & dados numéricos , Dor Aguda/tratamento farmacológico , Dor Aguda/prevenção & controle , Adulto , Feminino , Humanos , Mastectomia/efeitos adversos , Pessoa de Meia-Idade , Dor Pós-Operatória/tratamento farmacológico , Dor Pós-Operatória/prevenção & controle , Estudos RetrospectivosRESUMO
We adapted and tested a previously published questionnaire battery eliciting sensory and cognitive symptoms during (acute) and immediately after (post-acute) GHB intoxication. Studying 125 GHB users, we assessed the instrument's internal consistency using Cronbach's alpha (CA) and responsiveness to change comparing acute and post-acute symptoms. The final 14-item battery demonstrated good internal consistency (CA >or= 0.85, both acute and post-acute). The median symptom score (possible range 0-64) was 30 (acute) and 6 (post-acute; difference p < 0.001). This modified substance-specific symptom battery, which is easily administered, demonstrated excellent performance characteristics. It can be used to study GHB and, potentially, related drugs of abuse.
Assuntos
Adjuvantes Anestésicos/toxicidade , Afeto/efeitos dos fármacos , Cognição/efeitos dos fármacos , Desempenho Psicomotor/efeitos dos fármacos , Oxibato de Sódio/toxicidade , Transtornos Relacionados ao Uso de Substâncias/psicologia , Inquéritos e Questionários , Adulto , Feminino , Grupos Focais , Inquéritos Epidemiológicos , Humanos , Entrevistas como Assunto , Masculino , Motivação , Fumar/epidemiologia , Estados UnidosRESUMO
STUDY OBJECTIVE: To analyze changes in gamma-hydroxybutyrate (GHB) case reporting, we review GHB or congener drug cases reported to the California Poison Control System, comparing these to other data sets. METHODS: We identified cases from the California Poison Control System computerized database using standardized codes and key terms for GHB and congener drugs ("gamma butyrolactone," "1,4-butanediol," "gamma valerolactone"). We noted California Poison Control System date, caller and exposure site, patient age and sex, reported coingestions, and outcomes. We compared California Poison Control System data to case incidence from American Association of Poison Control Centers and Drug Abuse Warning Network data and drug use prevalence from National Institute for Drug Abuse survey data. RESULTS: A total of 1,331 patients were included over the 5-year period (1999-2003). California Poison Control System-reported GHB exposures decreased by 76% from baseline (n=426) to the final study year (n=101). The absolute decrease was present across all case types, although there was a significant proportional decrease in routine drug abuse cases and an increase in malicious events, including GHB-facilitated sexual assault (P=.002). American Association of Poison Control Centers data showed a similar decrease from 2001 to 2003. Drug Abuse Warning Network incidence flattened from 2001 to 2002 and decreased sharply in 2003. National Institute for Drug Abuse survey time trends were inconsistent across age groups. CONCLUSION: Based on the precipitous decrease in California Poison Control System case incidence for GHB during 5 years, the parallel trend in American Association of Poison Control Centers data, and a more recent decrease in Drug Abuse Warning Network cases, a true decrease in case incidence is likely. This could be due to decreased abuse rates or because fewer abusers seek emergency medical care. Case reporting may account for part of the decrease in the incidence of poison center contacts involving GHB.
Assuntos
Centros de Controle de Intoxicações/estatística & dados numéricos , Centros de Controle de Intoxicações/tendências , Oxibato de Sódio/intoxicação , Adolescente , Adulto , Distribuição por Idade , California/epidemiologia , Causalidade , Criança , Comorbidade , Bases de Dados Factuais , Feminino , Hospitalização/estatística & dados numéricos , Humanos , Masculino , Pessoa de Meia-Idade , Intoxicação/epidemiologia , Prevalência , Encaminhamento e Consulta/estatística & dados numéricos , Estudos Retrospectivos , Distribuição por Sexo , Transtornos Relacionados ao Uso de Substâncias/epidemiologia , Análise de SobrevidaRESUMO
STUDY OBJECTIVE: We sought to evaluate the safety and efficacy of a shorter N -acetylcysteine (NAC) regimen in the treatment of acute acetaminophen overdose. METHODS: We performed a retrospective case series in a large urban county hospital. Of 305 patients identified through the emergency department, 75 patients met the criteria inclusion: an acute overdose ingestion, serum acetaminophen concentration in toxic range according to the Rumack-Matthew nomogram, and oral NAC treatment initiated within 24 hours of the ingestion. The regional poison control center recommended oral treatment with NAC 140 mg/kg, followed by maintenance doses of 70 mg/kg every 4 hours until the serum acetaminophen level was no longer detectable, rather than the standard 72-hour treatment regimen. RESULTS: The primary outcome measure was the development of hepatotoxicity. Twenty-five (33.3%) patients were treated for a period of less than 24 hours, 25 (33.3%) were treated for 24 to 36 hours, and 25 (33.3%) were treated for 37 to 64 hours; the mean and median duration of treatment was 31 hours. None of the patients treated for less than 24 hours had evidence of hepatotoxicity (aspartate aminotransferase [AST] or alanine aminotransferase [ALT] level >1,000 IU/L); hepatotoxicity developed in 2 (8%) patients treated for 24 to 36 hours and 4 (16%) patients treated for 37 to 64 hours. There were no deaths or patients who received liver transplantation. The overall incidence of hepatotoxicity in our patients was similar to that found in other protocols with administration of oral NAC for 72 hours or intravenous NAC for 20 or 48 hours. CONCLUSION: This observational study suggests that a shorter course of oral NAC therapy in patients who do not show evidence of hepatotoxicity within 36 hours of an acute acetaminophen overdose is safe and effective. [Woo OF, Mueller PD, Olson KR, Anderson IB, Kim SY. Shorter duration of oral N -acetylcysteine therapy for acute acetami-nophen overdose. Ann Emerg Med . April 2000;35:363-368.].
RESUMO
AIM: To test the hypothesis that, in ELBW infants who did not receive antenatal MgSO4, lower baseline serum Mg is associated with poorer neurodevelopmental outcomes (NDO). STUDY DESIGN: The study was conducted in two phases: phase 1-- retrospective, and phase 2--prospective. SUBJECTS: Extremely low birth weight infants. OUTCOME MEASURES: Mortality and adverse NDO were assessed in relation to initial serum Mg measured in the first 12 hours of age. RESULTS: We studied 156 ELBW infants. In phase 1 (n=102): initial serum Mg (median [IQ range]) was greater in the infants who died compared to those who survived (1.7 [1.5-2.2] mg/dL vs. 1.6 [1.4-1.7] mg/dL, p=0.034). In phase 2 (n=54): initial serum Mg was greater in infants who died or had adverse NDO at 9 months when compared to those who survived with better NDO (1.7 [1.55-2.1] mg/dL vs. 1.5 [1.4-1.68] mg/dL, p=0.008). Using receiver operating characteristic (ROC) curve, increased Mg concentration in the first 12 hours>1.6 mg/dL was associated with unfavorable outcomes with sensitivity of 73%, specificity of 67%, and odds ratio of 5.5 (CI=1.2-24.8, p=0.037). CONCLUSIONS: In a cohort of preterm infants without antenatal exposure to MgSO4, initial serum Mg concentrations associated positively with poor outcomes. Further studies are needed in ELBW infants with poor NDO to determine whether they have a dysfunctional transport system that prevents Mg from entering into cells, or they have an active process that excretes Mg extracellularly.
Assuntos
Desenvolvimento Infantil , Recém-Nascido de Peso Extremamente Baixo ao Nascer/sangue , Recém-Nascido Prematuro/sangue , Magnésio/sangue , Estudos de Casos e Controles , Deficiências do Desenvolvimento/sangue , Deficiências do Desenvolvimento/diagnóstico , Feminino , Humanos , Recém-Nascido de Peso Extremamente Baixo ao Nascer/fisiologia , Recém-Nascido , Recém-Nascido Prematuro/fisiologia , Masculino , Doenças do Sistema Nervoso/sangue , Doenças do Sistema Nervoso/diagnóstico , Estudos ProspectivosRESUMO
INTRODUCTION: Little is known about behaviors linked to gamma hydroxybutyrate (GHB) morbidity. METHODS: We surveyed 131 GHB users, using logistic regression to test the associations between the high risk behaviors and hospital treatment for GHB (26 [20%] of subjects). RESULTS: Increased risk of GHB hospital treatment was associated with: co-ingestion of ethanol (OR 5.2; 95% CI 1.7-16), driving under the influence of GHB (OR 3.2; 95%, CI 1.3-7.8),use of GHB to treat withdrawal symptoms (OR 2.9; 95% CI 1.1-7.9), and co-ingestion of ketamine (OR 2.7; 95% CI 1.1-6.7). CONCLUSION: Targeted prevention activities could focus on selected high-risk behaviors.
Assuntos
Hospitalização/estatística & dados numéricos , Drogas Ilícitas/toxicidade , Assunção de Riscos , Oxibato de Sódio/toxicidade , Transtornos Relacionados ao Uso de Substâncias/epidemiologia , Adulto , Delirium por Abstinência Alcoólica/epidemiologia , Transtornos Relacionados ao Uso de Álcool/epidemiologia , Transtornos Relacionados ao Uso de Álcool/psicologia , Analgésicos , Condução de Veículo/psicologia , Condução de Veículo/estatística & dados numéricos , Comorbidade , Feminino , Inquéritos Epidemiológicos , Humanos , Ketamina , Masculino , Análise de Regressão , Transtornos Relacionados ao Uso de Substâncias/psicologia , Estados UnidosRESUMO
OBJECTIVE: There has been a recent proliferation of medical reference texts intended to guide practitioners whose patients use herbal therapies. We systematically assessed six herbal reference texts to evaluate the information they contain on herbal toxicity. METHODS: We selected six major herbal references published from 1996 to 2000 to evaluate the adequacy of their toxicological information in light of published adverse events. To identify herbs most relevant to toxicology, we reviewed herbal-related calls to our regional California Poison Control System, San Francisco division (CPCS-SF) in 1998 and identified the 12 herbs (defined as botanical dietary supplements) most frequently involved in these CPCS-SF referrals. We searched Medline (1966 to 2000) to identify published reports of adverse effects potentially related to these same 12 herbs. We scored each herbal reference text on the basis of information inclusiveness for the target 12 herbs, with a maximal overall score of 3. RESULTS: The herbs, identified on the basis of CPCS-SF call frequency were: St John's wort, ma huang, echinacea, guarana, ginkgo, ginseng, valerian, tea tree oil, goldenseal, arnica, yohimbe and kava kava. The overall herbal reference scores ranged from 2.2 to 0.4 (median 1.1). The Natural Medicines Comprehensive Database received the highest overall score and was the most complete and useful reference source. All of the references, however, lacked sufficient information on management of herbal medicine overdose, and several had incorrect overdose management guidelines that could negatively impact patient care. CONCLUSION: Current herbal reference texts do not contain sufficient information for the assessment and management of adverse health effects of botanical therapies.