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The lack of successful clinical trials in acute respiratory distress syndrome (ARDS) has highlighted the unmet need for biomarkers predicting ARDS mortality and for novel therapeutics to reduce ARDS mortality. We utilized a systems biology multi-"omics" approach to identify predictive biomarkers for ARDS mortality. Integrating analyses were designed to differentiate ARDS non-survivors and survivors (568 subjects, 27% overall 28-day mortality) using datasets derived from multiple 'omics' studies in a multi-institution ARDS cohort (54% European descent, 40% African descent). 'Omics' data was available for each subject and included genome-wide association studies (GWAS, n = 297), RNA sequencing (n = 93), DNA methylation data (n = 61), and selective proteomic network analysis (n = 240). Integration of available "omic" data identified a 9-gene set (TNPO1, NUP214, HDAC1, HNRNPA1, GATAD2A, FOSB, DDX17, PHF20, CREBBP) that differentiated ARDS survivors/non-survivors, results that were validated utilizing a longitudinal transcription dataset. Pathway analysis identified TP53-, HDAC1-, TGF-ß-, and IL-6-signaling pathways to be associated with ARDS mortality. Predictive biomarker discovery identified transcription levels of the 9-gene set (AUC-0.83) and Day 7 angiopoietin 2 protein levels as potential candidate predictors of ARDS mortality (AUC-0.70). These results underscore the value of utilizing integrated "multi-omics" approaches in underpowered datasets from racially diverse ARDS subjects.
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Síndrome do Desconforto Respiratório/mortalidade , Adulto , Idoso , Biomarcadores/análise , Biomarcadores/metabolismo , Metilação de DNA , Conjuntos de Dados como Assunto , Feminino , Perfilação da Expressão Gênica , Estudo de Associação Genômica Ampla , Humanos , Masculino , Pessoa de Meia-Idade , Proteômica , Síndrome do Desconforto Respiratório/sangue , Síndrome do Desconforto Respiratório/tratamento farmacológico , Síndrome do Desconforto Respiratório/genética , Medição de Risco/métodos , Análise de Sequência de RNA , Resultado do TratamentoRESUMO
INTRODUCTION: Patient flow is the process by which movement of patients and clinical productivity is achieved. The objectives of this study were to implement and evaluate the NHS Improvement SAFER patient flow bundle, evaluate the impact of the Red2Green initiative, and assess the impact of frailty on patient flow. MATERIALS AND METHODS: All patients admitted to a neurosurgery unit from 1 September to 30 November 2017 were included. Using guidance set out by NHS, data were prospectively collected from daily ward lists and patient notes, including demographics, admission and discharge details, length of stay, anticipated discharge date, red days with reasons and frailty (Rockwood Clinical Frailty Scale). NHS reference costs were used for cost analyses. RESULTS: A total of 420 patients (55% elective) were included, totalling 3909 bed days. All patients received daily senior reviews before midday, and anticipated discharge dates were set at daily multidisciplinary team meetings. Ten per cent of patients were discharged before midday. There were 21% (837) red days, significantly more (76%) for emergency patients (639 vs 198 elective; P < 0.001); 63% red days were attributed to awaiting a bed in a local hospital; 25% (106) patients were classed as frail (50 elective), which was associated with a significantly longer length of stay (17.3 vs 6; P < 0.01), and more red days (615 vs 222; p<0.01). Considering excess bed charges and lost revenue (with penalties), red days cost over £1 million per year. CONCLUSIONS: SAFER has identified areas for improvement in patient flow, with obvious cost implications. It has created a platform for discussion within the referral network and identified a role for a geriatric liaison service.
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Departamentos Hospitalares/estatística & dados numéricos , Procedimentos Neurocirúrgicos/estatística & dados numéricos , Aceitação pelo Paciente de Cuidados de Saúde/estatística & dados numéricos , Idoso , Ocupação de Leitos/estatística & dados numéricos , Procedimentos Cirúrgicos Eletivos/estatística & dados numéricos , Utilização de Instalações e Serviços , Feminino , Fragilidade/terapia , Humanos , Tempo de Internação/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Equipe de Assistência ao Paciente/estatística & dados numéricos , Alta do Paciente/estatística & dados numéricos , Segurança do Paciente , Estudos Prospectivos , Estudos Retrospectivos , Triagem/métodos , Triagem/estatística & dados numéricosRESUMO
We investigate the role of excited states in high-order harmonic generation by studying the spectral, spatial, and temporal characteristics of the radiation produced near the ionization threshold of argon by few-cycle laser pulses. We show that the population of excited states can lead either to direct extreme ultraviolet emission through free induction decay or to the generation of high-order harmonics through ionization from these states and recombination to the ground state. By using the attosecond lighthouse technique, we demonstrate that the high-harmonic emission from excited states is temporally delayed by a few femtoseconds compared to the usual harmonics, leading to a strong nonadiabatic spectral redshift.
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BACKGROUND: As an intraoperative imaging modality, ultrasound is a user-friendly and cost-effective real-time imaging technique. Despite this, it is still not routinely employed for brain tumor surgery. This may be due to the poor image quality in inexperienced hands, and the well-documented learning curve. However, with regular use, the operator issues are addressed, and intraoperative ultrasound can provide valuable real-time information. The aim of this review is to provide an understanding for neurosurgeons of the development and use of ultrasound in intracranial tumor surgery, and possible future advances. METHODS: A systematic search of the electronic databases Embase, Medline OvidSP, PubMed, Cochrane, and Google Scholar regarding the use of ultrasound in intracranial tumor surgery was undertaken. RESULTS AND DISCUSSION: Intraoperative ultrasound has been shown to be able to accurately account for brain shift and has potential for regular use in brain tumor surgery. Further developments in probe size, resolution, and image reconstruction techniques will ensure that intraoperative ultrasound is more accessible and attractive to the neuro-oncological surgeon. CONCLUSIONS: This review has summarized the development of ultrasound and its uses with particular reference to brain tumor surgery, detailing the ongoing challenges in this area.
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Neoplasias Encefálicas/cirurgia , Imageamento por Ressonância Magnética/métodos , Monitorização Intraoperatória/métodos , Procedimentos Neurocirúrgicos/métodos , Ultrassonografia/métodos , HumanosRESUMO
The integrity of the spinal accessory nerve is fundamental to thoracoscapular function and essential for scapulohumeral rhythm. This nerve is vulnerable along its superficial course. This study assessed the delay in diagnosis and referral for management of damage to this nerve, clarified its anatomical course and function, and documented the results of repair. From examination of our records, 111 patients with lesions of the spinal accessory nerve were treated between 1984 and 2007. In 89 patients (80.2%) the damage was iatropathic. Recognition and referral were seldom made by the surgeon responsible for the injury, leading to a marked delay in instituting treatment. Most referrals were made for painful loss of shoulder function. The clinical diagnosis is straightforward. There is a characteristic downward and lateral displacement of the scapula, with narrowing of the inferior scapulohumeral angle and loss of function, with pain commonly present. In all, 80 nerves were explored and 65 were repaired. The course of the spinal accessory nerve in relation to the sternocleidomastoid muscle was constant, with branches from the cervical plexus rarely conveying motor fibres. Damage to the nerve was predominantly posterior to this muscle. Despite the delay, the results of repair were surprising, with early relief of pain, implying a neuropathic source, which preceded generally good recovery of muscle function.
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Traumatismos do Nervo Acessório , Nervo Acessório/fisiopatologia , Nervo Acessório/cirurgia , Acidentes de Trânsito , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transferência de Nervo , Dor/etiologia , Nervos Periféricos/transplante , Recuperação de Função Fisiológica , Articulação do Ombro/fisiopatologia , Resultado do Tratamento , Violência , Adulto JovemRESUMO
Intraspinal re-implantation after traumatic avulsion of the brachial plexus is a relatively new technique. Three different approaches to the spinal cord have been described to date, namely the posterior scapular, anterolateral interscalenic multilevel oblique corpectomy and the pure lateral. We describe an anatomical study of the pure lateral approach, based on our clinical experience and studies on cadavers.
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Plexo Braquial/lesões , Plexo Braquial/cirurgia , Reimplante/métodos , Medula Espinal/cirurgia , Idoso , Idoso de 80 Anos ou mais , Plexo Braquial/anatomia & histologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/anatomia & histologia , Pescoço/anatomia & histologia , Posicionamento do Paciente/métodosRESUMO
Novel therapies are needed to address the vascular endothelial cell (EC) barrier disruption that occurs in inflammatory diseases such as acute lung injury (ALI). We previously demonstrated the potent barrier-enhancing effects of both sphingosine 1-phosphate (S1P) and the structurally similar compound FTY720 [2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol] in inflammatory lung injury. In this study, we examined the therapeutic potential of several novel FTY720 analogs to reduce vascular leak. Similar to S1P and FTY720, the (R)- and (S)-enantiomers of FTY720 phosphonate and enephosphonate analogs produce sustained EC barrier enhancement in vitro, as seen by increases in transendothelial electrical resistance (TER). In contrast, the (R)- and (S)-enantiomers of FTY720-regioisomeric analogs disrupt EC barrier integrity in a dose-dependent manner. Barrier-enhancing FTY720 analogs demonstrate a wider protective concentration range in vitro (1-50 microM) and greater potency than either S1P or FTY720. In contrast to FTY720-induced EC barrier enhancement, S1P and the FTY720 analogs dramatically increase TER within minutes in association with cortical actin ring formation. Unlike S1P, these FTY720 analogs exhibit differential phosphorylation effects without altering the intracellular calcium level. Inhibitor studies indicate that barrier enhancement by these analogs involves signaling via G(i)-coupled receptors, tyrosine kinases, and lipid rafts. Consistent with these in vitro responses, the (S)-phosphonate analog of FTY720 significantly reduces multiple indices of alveolar and vascular permeability in a lipopolysaccharide-mediated murine model of ALI (without significant alterations in leukocyte counts). These results demonstrate the capacity for FTY720 analogs to significantly decrease pulmonary vascular leakage and inflammation in vitro and in vivo.
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Permeabilidade Capilar/efeitos dos fármacos , Permeabilidade Capilar/fisiologia , Cloridrato de Fingolimode/análogos & derivados , Mediadores da Inflamação/síntese química , Mediadores da Inflamação/farmacologia , Organofosfonatos/síntese química , Organofosfonatos/farmacologia , Propilenoglicóis/síntese química , Propilenoglicóis/farmacologia , Artéria Pulmonar/efeitos dos fármacos , Esfingosina/análogos & derivados , Animais , Linhagem Celular , Cloridrato de Fingolimode/síntese química , Cloridrato de Fingolimode/farmacologia , Humanos , Pulmão/irrigação sanguínea , Pulmão/efeitos dos fármacos , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Artéria Pulmonar/patologia , Esfingosina/síntese química , Esfingosina/farmacologiaRESUMO
Our quality improvement program began in 2004 to improve cardiac surgery outcomes. Early tracheal extubation in the cardiovascular intensive unit was utilized as a multidisciplinary driver for the quality improvement program. Continuous improvement in the rate of early extubation to drive multidisciplinary quality improvement in cardiac critical care correlated with decreased mortality, morbidity, and improved operational efficiency. Supportive educational efforts included, but were not limited to, principles of change, trust, competing values, crew resource management, evidence based medicine, and quality improvement.
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INTRODUCTION: Early tracheal extubation is a common goal after cardiac surgery. Our study aims to examine whether timing of tracheal extubation predicts improved postoperative outcomes and late survival after cardiac surgery. We also evaluated the optimal timing of extubation and its association with better postoperative outcomes. METHODS: Between 2002 and 2006, 1164 patients underwent early tracheal extubation (<6 hours after surgery) and 1571 had conventional extubation (>6 hours after surgery). Propensity score adjustment and multivariable logistic regression analysis were used to adjust for imbalances in the patients' preoperative characteristics. Receiver operating characteristic curves (ROC) were used to identify the best timing of extubation and improved postoperative outcomes. Cox regression analysis was used to identify whether early extubation is a risk factor for decreased late mortality. RESULTS: Results - Early extubation was associated with lower propensity score-adjusted rate of operative mortality (Odds Ratio =0.55, 95% Confidence Intervals =0.31-0.98, p=0.043). Extubation within 9 hours emerged as the best predictor of improved postoperative morbidity and mortality (sensitivity =85.5%, specificity =52.7%, accuracy =64.5%). Early extubation also predicted decreased late mortality (Hazard Ratio =0.45, 95% Confidence Intervals 0.31-0.67, p<0.001). CONCLUSIONS: Early extubation may predict improved outcomes after cardiac surgery. Extubation within 9 hours after surgery was the best predictor of uncomplicated recovery after cardiac surgery. Those patients intubated longer than 16 hours have a poorer postoperative prognosis. Early extubation predicts prolonged survival up to 16 months after surgery.
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A 39 year-old farmer sustained a closed rupture of the left brachial artery, which was successfully managed by emergency vein graft repair of the artery. Adjacent nerve trunks were seen to be intact, but stretched. Burning pain in the distribution of the ulnar nerve started at day seven postoperatively, and worsened over the ensuing years. There was no response to membrane stabilising drugs, amitryptiline, nor to regional sympatholytic or local anaesthetic blocks. Neurolysis of the ulnar nerve, which was densely adherent to the dilated vein graft, abolished his pain.
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Dor Intratável/cirurgia , Nervo Ulnar , Adulto , Humanos , Masculino , Medição da Dor , Fatores de Tempo , Nervo Ulnar/cirurgiaRESUMO
Novel therapeutic strategies are needed to reverse the loss of endothelial cell (EC) barrier integrity that occurs during inflammatory disease states such as acute lung injury. We previously demonstrated potent EC barrier augmentation in vivo and in vitro by the platelet-derived phospholipid, sphingosine 1-phosphate (S1P) via ligation of the S1P1 receptor. The S1P analogue, FTY720, similarly exerts barrier-protective vascular effects via presumed S1P1 receptor ligation. We examined the role of the S1P1 receptor in sphingolipid-mediated human lung EC barrier enhancement. Both S1P and FTY-induced sustained, dose-dependent barrier enhancement, reflected by increases in transendothelial electrical resistance (TER), which was abolished by pertussis toxin indicating Gi-coupled receptor activation. FTY-mediated increases in TER exhibited significantly delayed onset and intensity relative to the S1P response. Reduction of S1P1R expression (via siRNA) attenuated S1P-induced TER elevations whereas the TER response to FTY was unaffected. Both S1P and FTY rapidly (within 5 min) induced S1P1R accumulation in membrane lipid rafts, but only S1P stimulated S1P1R phosphorylation on threonine residues. Inhibition of PI3 kinase activity attenuated S1P-mediated TER increases but failed to alter FTY-induced TER elevation. Finally, S1P, but not FTY, induced significant myosin light chain phosphorylation and dramatic actin cytoskeletal rearrangement whereas reduced expression of the cytoskeletal effectors, Rac1 and cortactin (via siRNA), attenuated S1P-, but not FTY-induced TER elevations. These results mechanistically characterize pulmonary vascular barrier regulation by FTY720, suggesting a novel barrier-enhancing pathway for modulating vascular permeability.
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Células Endoteliais/efeitos dos fármacos , Imunossupressores/farmacologia , Propilenoglicóis/farmacologia , Receptores de Lisoesfingolipídeo/metabolismo , Esfingosina/análogos & derivados , Adenoviridae/genética , Permeabilidade Capilar , Células Cultivadas , Citoesqueleto/metabolismo , Impedância Elétrica , Endotélio Vascular/citologia , Cloridrato de Fingolimode , Humanos , Pulmão/citologia , Modelos Biológicos , Fosforilação , Artéria Pulmonar/citologia , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Esfingosina/farmacologia , Treonina/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismoRESUMO
There are few longitudinal studies of cognition in patients with multiple sclerosis, and the results of these studies remain inconclusive. No serial neuropsychological data of an exclusively primary progressive series are available. Cross-sectional analyses have revealed significant correlations between cognition and magnetic resonance imaging (MRI) parameters in primary progressive multiple sclerosis (PPMS). This study investigated cognitive and MRI change in 99 PPMS patients from five European centres for 2 years. They were assessed at 12 month intervals using the Brief Repeatable Battery, a reasoning test, and a measure of depression. The MRI parameters of T1 hypointensity load, T2 lesion load, and partial brain volume were also calculated at each time point. There were no significant differences between the mean cognitive scores of the patients at year 0 and year 2. However, one-third of the patients demonstrated absolute cognitive decline on individual test scores. Results indicated that initial cognitive status on entry into the study was a good predictor of cognitive ability at 2 years. There was only a small number of significant correlations between changes in cognition and changes on MRI, notably T1 hypointensity load with the two attentional tasks (r = -0.266, P = 0.017; r = -0.303, P = 0.012). It is probable that multiple factors underlie this weak relation between the cognitive and MRI measures.
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Transtornos Cognitivos/psicologia , Esclerose Múltipla/psicologia , Adulto , Encéfalo/patologia , Transtornos Cognitivos/patologia , Progressão da Doença , Feminino , Humanos , Estudos Longitudinais , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/patologia , Testes Neuropsicológicos , Escalas de Graduação Psiquiátrica , Testes Psicológicos , Estatísticas não ParamétricasRESUMO
TorsinA is a novel protein identified in the search for mutations underlying the human neurologic movement disorder, early onset torsion dystonia. Relatively little is understood about the normal function of torsinA or the physiological effects of the codon deletion associated with most cases of disease. Overexpression of wild-type torsinA in cultured cells by DNA transfection results in a reticular distribution of immunoreactive protein that co-localizes with endoplasmic reticulum resident chaperones, while the dystonia-related mutant form accumulates within concentric membrane whorls and nuclear-associated membrane stacks. In this study we examined the biogenesis of mutant torsinA-positive membrane inclusions using tetracycline-regulated herpes simplex virus amplicon vectors. At low expression levels, mutant torsinA was localized predominantly around the nucleus, while at high levels it was also concentrated within cytosolic spheroid inclusions. In contrast, the distribution of wild-type torsinA did not vary, appearing diffuse and reticular at all expression levels. These observations are consistent with descriptions of inducible membrane synthesis in other systems in which cytosolic membrane whorls are derived from multilayered membrane stacks that first form around the nuclear envelope. These results also suggest that formation of mutant torsinA-positive inclusions occurs at high expression levels in culture, whereas the perinuclear accumulation of the mutant protein is present even at low expression levels that are more likely to resemble those of the endogenous protein. These nuclear-associated membrane structures enriched in mutant torsinA may therefore be of greater relevance to understanding how the dystonia-related mutation compromises cellular physiology.
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Proteínas de Transporte/metabolismo , Núcleo Celular/metabolismo , Corpos de Inclusão/metabolismo , Membranas Intracelulares/metabolismo , Chaperonas Moleculares/metabolismo , Organelas/metabolismo , Animais , Biomarcadores , Proteínas de Transporte/genética , Linhagem Celular , Núcleo Celular/genética , Núcleo Celular/patologia , Citosol/metabolismo , Citosol/patologia , Distonia Muscular Deformante/genética , Distonia Muscular Deformante/metabolismo , Distonia Muscular Deformante/fisiopatologia , Genes Reporter/genética , Vetores Genéticos/genética , Herpes Simples/genética , Humanos , Corpos de Inclusão/genética , Corpos de Inclusão/patologia , Membranas Intracelulares/patologia , Chaperonas Moleculares/genética , Mutação/genética , Membrana Nuclear/metabolismo , Membrana Nuclear/patologia , Organelas/genética , Organelas/patologia , Tetraciclina/farmacologia , Transgenes/genéticaRESUMO
Removing the prescription requirement for Plan B will help ensure that the product plays a larger role nationally in the reduction of unintended pregnancy and abortion-important public health goals. Over-the-counter (OTC) sale of Plan B should present no serious safety issues. OTC consumers are able to understand and follow the instructions for proper use of Plan B. Efficacy of the OTC product is likely to be the same as, or better than, the prescription product, given more timely access to treatment. Based on the results of a growing body of literature and foreign marketing experience, the risk of unintended health consequences also appears to be minimal. There is no evidence to suggest that American women will abuse Plan B as an OTC product.
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Anticoncepcionais Orais Sintéticos/provisão & distribuição , Anticoncepcionais Pós-Coito/provisão & distribuição , Levanogestrel/provisão & distribuição , Medicamentos sem Prescrição/provisão & distribuição , Feminino , Humanos , Legislação de Medicamentos , Assistência Farmacêutica , Medição de Risco , Estados UnidosRESUMO
Herpes simplex virus type 1/adeno-associated virus (HSV/AAV) rep(+) hybrid amplicon vectors containing AAV inverted terminal repeats (ITRs) and rep gene sequences can mediate site-specific integration into the human genome. In this study, we have generated and characterized the first transgenic mice that bear the full-length (8.2 kb) human AAVS1 locus. Immortalized mouse embryonic fibroblasts from this mouse line were transduced with the rep(+), rep(-) (containing only ITRs flanking the transgene) hybrid amplicon vectors, and the standard amplicon vector to determine stable integration frequency and the site of integration. Transduction of transgenic fibroblasts resulted in a 10-fold higher stable integration frequency with rep(+) hybrid amplicon vector than with rep(-) or standard amplicon vectors. Southern blot analysis of genomic DNA from transgenic cells stably transduced with the rep(+) hybrid amplicon vector revealed site-specific integration of transgenes at the AAVS1 locus in 50% of clones. Some site-specific and random integration events were limited to the ITR-flanked transgene cassette. In contrast, transduction of transgenic mouse cells with the rep(-) or standard amplicon vectors resulted in random integrations of the entire rep(-) hybrid amplicon or amplicon DNA that were incorporated into the host genome as a concatenate of various sizes. These results demonstrate for the first time that the genome of transgenic mice bearing the human AAVS1 locus serves as a platform for site-specific integration of AAV ITR-flanked transgene cassettes within the hybrid amplicon vector in the presence of Rep.
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Dependovirus/genética , Fibroblastos/metabolismo , Terapia Genética/métodos , Vetores Genéticos/genética , Simplexvirus/genética , Animais , Linhagem Celular Transformada , Engenharia Genética , Vetores Genéticos/administração & dosagem , Humanos , Camundongos , Camundongos Endogâmicos , Camundongos Transgênicos , Mutagênese Sítio-Dirigida , Transdução Genética/métodosRESUMO
Enrofloxacin was administered i.v. to five adult mares at a dose of 5 mg/kg. After administration, blood and endometrial biopsy samples were collected at regular intervals for 24 h. The plasma and tissue samples were analyzed for enrofloxacin and the metabolite ciprofloxacin by high-pressure liquid chromatography. In plasma, enrofloxacin had a terminal half-life (t(1/2)), volume of distribution (area method), and systemic clearance of 6.7 +/- 2.9 h, 1.9 +/- 0.4 L/kg, and 3.7 +/- 1.4 mL/kg/min, respectively. Ciprofloxacin had a maximum plasma concentration (Cmax) of 0.28 +/- 0.09 microg/mL. In endometrial tissue, the enrofloxacin Cmax was 1.7 +/- 0.5 microg/g, and the t(1/2) was 7.8 +/- 3.7 h. Ciprofloxacin Cmax in tissues was 0.15 +/- 0.04 microg/g and the t(1/2) was 5.2 +/- 2.0 h. The tissue:plasma enrofloxacin concentration ratios (w/w:w/v) were 0.175 +/- 0.08 and 0.47 +/- 0.06 for Cmax and AUC, respectively. For ciprofloxacin, these values were 0.55 +/- 0.13 and 0.58 +/- 0.31, respectively. We concluded that plasma concentrations achieved after 5 mg/kg i.v. are high enough to meet surrogate markers for antibacterial activity (Cmax:MIC ratio, and AUC:MIC ratio) considered effective for most susceptible gram-negative bacteria. Endometrial tissue concentrations taken from the mares after dosing showed that enrofloxacin and ciprofloxacin both penetrate this tissue adequately after systemic administration and would attain concentrations high enough in the tissue fluids to treat infections of the endometrium caused by susceptible bacteria.
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Anti-Infecciosos/farmacocinética , Ciprofloxacina/farmacocinética , Endométrio/metabolismo , Fluoroquinolonas , Cavalos/metabolismo , Quinolonas/farmacocinética , Animais , Anti-Infecciosos/administração & dosagem , Anti-Infecciosos/sangue , Área Sob a Curva , Cromatografia Líquida de Alta Pressão/veterinária , Ciprofloxacina/administração & dosagem , Ciprofloxacina/sangue , Enrofloxacina , Feminino , Infusões Intravenosas/veterinária , Quinolonas/administração & dosagem , Quinolonas/sangueRESUMO
Herpes simplex virus type 1 (HSV-1) amplicon vectors are promising gene delivery tools, but their utility in gene therapy has been impeded to some extent by their inability to achieve stable transgene expression. In this study, we examined the possibility of improving transduction stability in cultured human cells via site-specific genomic integration mediated by adeno-associated virus (AAV) Rep and inverted terminal repeats (ITRs). A rep(-) HSV/AAV hybrid amplicon vector was made by inserting a transgene cassette flanked with AAV ITRs into an HSV-1 amplicon backbone, and a rep(+) HSV/AAV hybrid amplicon was made by inserting rep68/78 outside the rep(-) vector 3' AAV ITR sequence. Both vectors also had a pair of loxP sites flanking the ITRs. The resulting hybrid amplicon vectors were successfully packaged and compared to a standard amplicon vector for stable transduction frequency (STF) in human 293 and Gli36 cell lines and primary myoblasts. The rep(+), but not the rep(-), hybrid vector improved STF in all three types of cells; 84% of Gli36 and 40% of 293 stable clones transduced by the rep(+) hybrid vector integrated the transgene into the AAVS1 site. Due to the difficulty in expanding primary myoblasts, we did not assess site-specific integration in these cells. A strategy to attempt further improvement of STF by "deconcatenating" the hybrid amplicon DNA via Cre-loxP recombination was tested, but it did not increase STF. These data demonstrate that introducing the integrating elements of AAV into HSV-1 amplicon vectors can significantly improve their ability to achieve stable gene transduction by conferring the AAV-like capability of site-specific genomic integration in dividing cells.
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Proteínas de Ligação a DNA/genética , Dependovirus/genética , Vetores Genéticos , Herpesvirus Humano 1/genética , Transdução Genética , Transgenes , Proteínas Virais/genética , Linhagem Celular , Proteínas de Ligação a DNA/metabolismo , Dependovirus/metabolismo , Amplificação de Genes , Expressão Gênica , Técnicas de Transferência de Genes , Herpesvirus Humano 1/metabolismo , Humanos , Recombinação Genética , Sequências Repetidas Terminais/genética , Proteínas Virais/metabolismo , Integração ViralRESUMO
Memory tests were often developed for healthy populations. The accuracy of these measures is reduced when administered to patients with neurological diseases, who may experience physical and/or cognitive symptoms. Also, methodological factors, for example, spanning the ability spectrum, and content/format artefacts, may contribute to a decline in test precision. The aim of this study was to develop a new test of memory, which addresses these issues. The new memory test comprises assessments of recall, paired association, and recognition, at a Task Familiarisation stage and two difficulty levels, for both the verbal and spatial modalities. It was administered to 85 healthy individuals and 100 patients with multiple sclerosis (MS). All patients were able to attempt each task of the new assessment and there was no influence of visual integrity or manual dexterity on memory test performance, supporting the applicability of the tasks to patients with multiple sclerosis. Both the standardisation and validation samples demonstrated a wide range of scores on each section of the new test suggesting that the measure spanned an acceptably broad range of abilities. It seems probable, therefore, that the new assessment offers a more exact measure of verbal and spatial recall, paired association, and recognition memory.
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Memória de Curto Prazo/fisiologia , Esclerose Múltipla/psicologia , Testes Psicológicos , Adulto , Idoso , Ansiedade , Cognição , Depressão/classificação , Inglaterra , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reconhecimento Psicológico , Valores de Referência , Reprodutibilidade dos Testes , Classe Social , Percepção Espacial , Fala , DesempregoRESUMO
OBJECTIVE: The effectiveness of federal prison-based residential drug and alcohol treatment programs was evaluated using event history procedures that addressed the problem of selection bias and included a wide range of control variables. METHODS: The sample comprised 760 treatment subjects and 809 comparison subjects. Treatment subjects were from 20 different prisons of medium, low, and minimum security levels. Comparison subjects were drawn from over 30 prisons. RESULTS: The results indicated that individuals who entered and completed in-prison residential treatment were less likely to experience the critical postrelease outcomes of new arrests and substance use during the first 6 months following release. CONCLUSIONS: Without controlling for selection bias, the effects of treatment would most likely have been attenuated. The results have greater generalizability than other studies of prison-based treatment. This study occurred within a multisite context of 20 programs serving both male and female inmates and operating within different security levels and different geographic regions.
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Internação Compulsória de Doente Mental , Prisões , Tratamento Domiciliar , Transtornos Relacionados ao Uso de Substâncias/epidemiologia , Transtornos Relacionados ao Uso de Substâncias/reabilitação , Adulto , Feminino , Humanos , Incidência , Masculino , Índice de Gravidade de Doença , Transtornos Relacionados ao Uso de Substâncias/diagnóstico , Fatores de Tempo , Resultado do TratamentoRESUMO
In an evaluation of prison-based residential drug treatment programs, the authors use three different regression-based approaches to estimating treatment effects. Two of the approaches, the instrumental variable and the Heckman approach, attempt to minimize selection bias as an explanation for treatment outcomes. Estimates from these approaches are compared with estimates from a regression in which treatment is represented by a dummy variable. The article discusses the advantage of using more than one method to increase confidence in findings when possible selection bias is a concern. Three-year outcome data for 2,315 federal inmates are used in analyses where the authors separately examine criminal recidivism and relapse to drug use for men and women. Statistical tests lead the authors to conclude that treatment reduces criminal recidivism and relapse to drug use. The treatment effect was largest when the inference was based on the Heckman approach, somewhat smaller when based on the instrumental variable approach, and smallest when based on the traditional dummy variable approach. Treatment effects for females were not statistically significant.