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Asma , Citocinas , Humanos , Citocinas/metabolismo , Alarminas/metabolismo , Esputo/metabolismo , Asma/diagnóstico , Asma/metabolismo , FenotipoRESUMEN
BACKGROUND: It remains unclear whether patients with asthma and/or chronic obstructive pulmonary disease (COPD) are at increased risk for severe coronavirus disease 2019 (COVID-19). OBJECTIVE: Compare in-hospital COVID-19 outcomes among patients with asthma, COPD, and no airway disease. METHODS: A retrospective cohort study was conducted on 8,395 patients admitted with COVID-19 between March 2020 and April 2021. Airway disease diagnoses were defined using International Classification of Diseases, 10th Revision codes. Mortality and sequential organ failure assessment (SOFA) scores were compared among groups. Logistic regression analysis was used to identify and adjust for confounding clinical features associated with mortality. RESULTS: The median SOFA score in patients without airway disease was 0.32 and mortality was 11%. In comparison, asthma patients had lower SOFA scores (median 0.15; P < .01) and decreased mortality, even after adjusting for age, diabetes, and other confounders (odds ratio 0.65; P = .01). Patients with COPD had higher SOFA scores (median 0.86; P < .01) and increased adjusted odds of mortality (odds ratio 1.40; P < .01). Blood eosinophil count of 200 cells/µL or greater, a marker of type 2 inflammation, was associated with lower mortality across all groups. Importantly, patients with asthma showed improved outcomes even after adjusting for eosinophilia, indicating that noneosinophilic asthma was associated with protection as well. CONCLUSIONS: COVID-19 severity was increased in patients with COPD and decreased in those with asthma, eosinophilia, and noneosinophilic asthma, independent of clinical confounders. These findings suggest that COVID-19 severity may be influenced by intrinsic immunological factors in patients with airway diseases, such as type 2 inflammation.
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Asma , COVID-19 , Diabetes Mellitus Tipo 2 , Eosinofilia , Enfermedad Pulmonar Obstructiva Crónica , Humanos , Estudios Retrospectivos , COVID-19/complicaciones , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Asma/diagnóstico , Inflamación , Eosinofilia/complicacionesRESUMEN
Whether or not to give research results back to individuals whose specimens are used for biomedical research is a subject of considerable controversy. Much of the debate has been focused around the ethical and legal concerns with some consideration of broader social issues such as whether or not people will be affected by such information for employment or health care. Much less attention has been paid to biobanks that collect the specimens used to generate the research findings and the issues and operational requirements for implementing return of individual research results. In this article, we give the biobanks' perspective and highlight that given the diversity among the types of biobanks, it may be difficult to design and implement a blanket policy in this complex area. We discuss the variability in the types of biobanks and some important issues that should be considered in determining whether or not research results should be provided to individuals whose specimens are used in biomedical research. We also discuss challenges that should be considered in implementing any approaches to the return of research results.
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Investigación Biomédica/ética , Informática Médica/ética , Sujetos de Investigación , Bancos de Tejidos/ética , Investigación Biomédica/métodos , Investigación Biomédica/estadística & datos numéricos , Humanos , Informática Médica/métodos , Informática Médica/estadística & datos numéricos , Relaciones Investigador-Sujeto/ética , Revelación de la Verdad/éticaRESUMEN
Fluorescence intensities of propranolol and atenolol in binary solvent mixtures at various temperatures are measured and mathematical models are proposed to represent the fluorescence intensity data. The results showed that the proposed models are able to correlate/predict the data with reasonable error. The fluorescence intensity of pyridoxal HCl in binary solvents at 25 °C is also determined and represented by the proposed model as an additional test probe.
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Simulación por Computador , Fluorescencia , Colorantes Fluorescentes/química , Solventes/química , Alcoholes/química , Solubilidad , Termodinámica , Agua/químicaRESUMEN
PURPOSE: The aim of this study was to identify the dominant factors affecting the stability of nanoemulsions, using artificial neural networks (ANNs). METHODS: A nanoemulsion preparation of budesonide containing polysorbate 80, ethanol, medium chain triglycerides and saline solution was designed, and the particle size of samples with various compositions, prepared using different rates and amounts of applied ultrasonic energy, was measured 30 min and 30 days after preparation. Using ANNs, data were modelled and assessed. The derived predictive model was validated statistically and then used to determine the effect of different formulation and processing input variables on particle size growth of the nanoemulsion preparation as an indicator of the preparation stability. RESULTS: The results indicated that the data can be satisfactorily modelled using ANNs, while showing a high degree of complexity between the dominant factors affecting the stability of the preparation. CONCLUSION: The total amount of applied energy and concentration of ethanol were found to be the dominant factors controlling the particle size growth.
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Budesonida/química , Emulsiones/química , Nanotecnología/métodos , Redes Neurales de la Computación , Composición de Medicamentos/métodos , Estabilidad de Medicamentos , Etanol/química , Tamaño de la Partícula , Polisorbatos/química , Cloruro de Sodio/química , Factores de Tiempo , Triglicéridos/química , UltrasonidoRESUMEN
The aim of this work was to evaluate the in vitro performance of a nebulized nanoemulsion formulation which had been optimised previously. To do so, a transparent nanoemulsion preparation containing 1.5 mg/ml of budesonide was prepared and diluted to achieve concentrations of 250 and 500 µg/ml budesonide. The in vitro characteristics of the diluted nanoemulsions were then compared with the commercially available suspension of budesonide (Pulmicort Respules®) when nebulized using a jet and a vibrating mesh nebulizer. A smaller MMAD with improved aerosol output was observed in the nanoemulsion preparations compared with the corresponding suspension formulations indicating an improved in vitro performance for the nanoemulsion-based preparations.
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Aerosoles/administración & dosificación , Aerosoles/química , Emulsiones/síntesis química , Nanopartículas/química , Nebulizadores y Vaporizadores , Fármacos del Sistema Respiratorio/administración & dosificación , Fármacos del Sistema Respiratorio/química , Administración por Inhalación , Composición de Medicamentos/métodos , Ensayo de Materiales , Nanopartículas/administración & dosificaciónRESUMEN
A new rapid, sensitive and validated HPLC method has been developed for the determination of methylxanthines and their metabolites in asthmatic patients. The method was initiated by using spiked urine samples on a silica monolithic column as a novel packing material. The mobile phase consisted of 10 mM potassium dihydrogen phosphate buffer/methanol (87.5:12.5 v/v), at a flow rate 1 mL/min. Detection was set at 274 nm. The LOQ for all the compounds ranged from 14 to 41 ng/mL. Excellent linearity was achieved over the studied range of concentration with correlation coefficients 0.9991-0.9998 (n = 6). The developed method was validated by precision and accuracy with RSD <2.55%. On extraction of the drugs and metabolites from the urine samples high recoveries were achieved ranging from 82.06 to 98.34% w/w on RP18 cartridges and methanol/chloroform (20:80 v/v) as the extraction solvent. This method has advantages over other methods using conventional C18 packings.
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Cromatografía Líquida de Alta Presión/instrumentación , Cromatografía Líquida de Alta Presión/métodos , Orina/química , Xantinas/análisis , Xantinas/metabolismo , Humanos , Modelos Lineales , Estructura Molecular , Reproducibilidad de los Resultados , Extracción en Fase Sólida , Factores de Tiempo , Xantinas/químicaRESUMEN
Herein we report the results of a comparative study on the performance of Monolithic RP-18e and Platinum C(18) 3 mum columns for isocratic separation of acidic and basic test compounds. The inter- and intraday precision of different practical parameters including number of theoretical plates (N), capacity factor (K'), tailing factor (T(0.05)), and resolution (R(s)) were determined for both columns. Two different production batches were used for each column and batch to batch reproducibility of both columns was evaluated. The column backpressure drop over flow rate range 0.5-2 mL/min at the monolithic columns was two- to three-times lower than that on the platinum column without loss of the column efficiency. The plate heights were used to estimate the columns efficiency using Van Deemter plots. Both types of columns were able to separate the tested compounds well with sufficient resolution and peak symmetry but they differ in the analysis time and column backpressure, significantly. Monolithic column was more convenient as it enables the analytical run under low backpressure at shorter time with sufficient separation efficiency.
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Ácidos/análisis , Cromatografía Líquida de Alta Presión/instrumentación , Cromatografía Líquida de Alta Presión/métodos , Concentración de Iones de Hidrógeno , Estructura Molecular , Preparaciones Farmacéuticas/análisis , Preparaciones Farmacéuticas/química , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The purpose of this study was to use Artificial Neural Networks (ANNs) in identifying factors, in addition to surfactant and internal phase content, that influence the particle size of nanoemulsions. The phase diagram and rheometric characteristics of a nanoemulsion system containing polysorbate 80, ethanol, medium chain triglycerides and normal saline loaded with budesonide were investigated. The particle size of samples of various compositions prepared using different rates and amounts of applied energy was measured. Data, divided into training, test and validation sets, were modelled by ANNs. The developed model was assessed and found to be of high quality. The model was then used to explore the effect of composition and processing factors on particle size of the nanoemulsion preparation. The study demonstrates the potential of ANNs in identifying critical parameters controlling preparation for this system, with the total amount of applied energy during preparation found to be the dominant factor in controlling the final particle size.
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Emulsiones , Nanopartículas , Algoritmos , Antiinflamatorios/administración & dosificación , Antiinflamatorios/química , Budesonida/administración & dosificación , Budesonida/química , Interpretación Estadística de Datos , Etanol/química , Excipientes , Indicadores y Reactivos , Redes Neurales de la Computación , Tamaño de la Partícula , Polisorbatos , Reproducibilidad de los Resultados , Programas Informáticos , Solventes , Triglicéridos/administración & dosificación , Triglicéridos/químicaRESUMEN
To determine the usefulness of polymerase chain reaction (PCR) analyses in the diagnosis of lymphoid infiltrate cells in ocular samples, PCR was performed using oligonucleotide primers specific for immunoglobulin heavy chain rearrangement at framework 2, framework 3, and t(14;18) translocation of the bcl-2 gene. These were used to successfully generate amplicons of 220 to 230 bp, 110 to 120 bp, and 175 to 200 bp, respectively. After PCR amplification, primers directed against the t(14;18) detected 10 pg of B-cell lymphoma DNA. PCR against Fr2 and Fr3 IgH rearrangement detected 10 fg and 10 pg in the seminested PCR, respectively. Conventional pathological methods were highly accurate at establishing the correct final diagnosis in formalin-fixed, paraffin-embedded samples but were much less sensitive and predictive in cytological specimens of intraocular fluid. A combination of the three PCR reactions was an equally successful diagnostic approach on paraffin-embedded samples, whereas single PCR reactions did not significantly improve diagnosis over histopathological diagnostic techniques. Thus, a combination of PCR reactions is useful in the detection of B-cell monoclonality, aids the differentiation between lymphomatous and inflammatory infiltrates, and is more powerful as a diagnostic method than single PCR or conventional cytopathology for lymphoid infiltrates in ocular fluid aspirates.
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Humor Acuoso/citología , Neoplasias del Ojo/genética , Linfoma de Células B Grandes Difuso/genética , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Cartilla de ADN , Estudios de Evaluación como Asunto , Reordenamiento Génico de Linfocito B/genética , Genes bcl-2/genética , Humanos , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Translocación Genética/genéticaRESUMEN
The difficulties with 'retained organs' in the UK have resulted in a new legislation relating to human organs, tissues, and bodies - the Human Tissue Act 2004 and the Human Tissue Act Scotland 2006 are now in place. The new laws apply to a wide range of activities including transplantation, education, clinical audit, the practice of autopsies, anatomical examination and others, including the use of human tissues in research. Pathobiology research that uses human tissues is now undertaken in a regulated environment in the UK. The details of these regulations are described and the consequences discussed. In the second part of the paper the patient's views and expectations in this new setting are forwarded.
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Investigación Biomédica/legislación & jurisprudencia , Satisfacción del Paciente , Bancos de Tejidos/legislación & jurisprudencia , Investigación Biomédica/ética , Humanos , Consentimiento Informado , Bancos de Tejidos/ética , Reino UnidoRESUMEN
Quantitative gene expression data are often normalized to the expression levels of control or so-called "housekeeping" genes. An inherent assumption in the use of housekeeping genes is that expression of the genes remains constant in the cells or tissues under investigation. Although exceptions to this assumption are well documented, housekeeping genes are of value in fully characterized systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is one of the most commonly used housekeeping genes used in comparisons of gene expression data. To investigate the value of GAPDH as a housekeeping gene in human tissues, the expression of GAPDH mRNA was measured in a panel of 72 different pathologically normal human tissue types. Measurements were obtained from 371,088 multiplexed, quantitative real-time RT-PCRs with specific target genes. Significant differences in the expression levels of GAPDH mRNA were observed between tissue types and between donors of the same tissue. A 15-fold difference in GAPDH mRNA copy numbers was observed between the highest and lowest expressing tissue types, skeletal muscle and breast, respectively. No specific effect of either age or gender was observed on GAPDH mRNA expression. These data provide an extensive analysis of GAPDH mRNA expression in human tissues and confirm previous reports of the marked variability of GAPDH expression between tissue types. These data establish comparative levels of expression and can be used to add value to gene expression data in which GAPDH is used as the internal control.
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Gliceraldehído-3-Fosfato Deshidrogenasas/genética , ARN Mensajero/genética , Actinas/genética , Cadáver , Femenino , Regulación Enzimológica de la Expresión Génica , Humanos , Masculino , Especificidad de Órganos , ARN Mensajero/aislamiento & purificación , Receptores de Transferrina/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción GenéticaRESUMEN
Phototransduction in Drosophila is a phosphoinositide-mediated signalling pathway. Phosphatidylinositol 4,5-bisphosphate (PIP2) plays a central role in this process, and its levels are tightly regulated. A photoreceptor-specific form of the enzyme CDP-diacylglycerol synthase (CDS), which catalyzes the formation of CDP-diacylglycerol from phosphatidic acid, is a key regulator of the amount of PIP2 available for signalling. cds mutants develop light-induced retinal degeneration. We report here the isolation and characterization of two murine genes encoding this enzyme, Cds1 and Cds2. The genes encode proteins that are 73% identical and 92% similar but exhibit very different expression patterns. Cds1 shows a very restricted expression pattern but is expressed in the inner segments of the photoreceptors whilst Cds2 shows a ubiquitous pattern of expression. Using fluorescent in situ hybridization we have mapped Cds1 and Cds2 to chromosomes 5E3 and 2G1 respectively. These are regions of synteny with the corresponding human gene localization (4q21 and 20p13). Transient transfection experiments with epitope tagged proteins have also demonstrated that both are associated with the endoplasmic reticulum.
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Diacilglicerol Colinafosfotransferasa/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Células CHO , Mapeo Cromosómico , Cromosomas de los Mamíferos/genética , Cricetinae , Cricetulus , ADN Complementario/química , ADN Complementario/genética , ADN Complementario/aislamiento & purificación , Diacilglicerol Colinafosfotransferasa/metabolismo , Retículo Endoplásmico/metabolismo , Exones , Femenino , Perfilación de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Genes/genética , Hibridación Fluorescente in Situ , Intrones , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Ratones , Microscopía Confocal , Microscopía Fluorescente , Datos de Secuencia Molecular , Plásmidos/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , TransfecciónRESUMEN
PURPOSE: The Leber congenital amaurosis (LCA) protein AIPL1 is present only in the rod photoreceptors of the adult human retina and is excluded from the cone photoreceptors. LCA, however, is characterized by an absence of both rod and cone function at birth or shortly thereafter. Therefore, this study was conducted to determine whether AIPL1 is present in the rod and cone photoreceptors of the developing human retina. In addition, the expression of NUB1, a putative AIPL1-interacting partner, was examined. METHODS: A comprehensive spatiotemporal examination of AIPL1 distribution during development was performed by immunohistochemistry, using a previously characterized AIPL1 anti-serum. Immunofluorescence confocal microscopy was used to examine the coexpression of AIPL1 with the long/medium (L/M) and short (S) wavelength-sensitive cone photoreceptors in the developing human retina. The spatiotemporal distribution of NUB1 was also examined by immunohistochemistry, using a newly developed anti-serum to the C terminus of NUB1. RESULTS: AIPL1 protein was detected by 11.8 fetal weeks in the central fetal human retina. With continued development, AIPL1 expression spread gradually toward peripheral retina. AIPL1 was expressed in the L/M and S cone photoreceptors in addition to the rods of the developing human retina. NUB1 was localized in cell nuclei throughout the human fetal and adult eye at all time points. CONCLUSIONS: The pattern of AIPL1 expression closely follows the centroperipheral gradient in photoreceptor development. The data suggest that AIPL1 is essential for the normal development of both rod and cone photoreceptor cells and that mutations in the AIPL1 gene cause the death or dysfunction of photoreceptors early in development resulting in blindness or severely impaired vision at birth.
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Células Fotorreceptoras Retinianas Conos/embriología , Células Fotorreceptoras Retinianas Bastones/embriología , Proteínas Adaptadoras Transductoras de Señales , Anciano , Western Blotting , Proteínas Portadoras , Desarrollo Embrionario y Fetal , Proteínas del Ojo , Edad Gestacional , Humanos , Inmunohistoquímica , Microscopía Confocal , Microscopía Fluorescente , Factores de Transcripción/metabolismoRESUMEN
PURPOSE: To investigate whether 173 patients with microphthalmia, anophthalmia, and coloboma have mutations in the eye-development gene SIX6. METHODS: The two exons of the SIX6 gene were amplified by PCR from patients' genomic DNA and directly sequenced to search for mutations. The PCR products of 75 patients were also analyzed by denaturing high-performance liquid chromatography (DHPLC). RESULTS: Six SIX6 polymorphisms were identified in the patient panel. Three of these polymorphisms change the encoded amino acid. However, all six polymorphisms were also identified in unaffected individuals. There was no statistically significant difference in genotypes between patients and control subjects. CONCLUSIONS: No evidence was found that SIX6 mutations underlie human congenital structural eye malformations.
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Anoftalmos/genética , Coloboma/genética , Proteínas del Ojo/genética , Proteínas de Homeodominio/genética , Microftalmía/genética , Mutación , Transactivadores/genética , Adulto , Cromatografía Líquida de Alta Presión , Análisis Mutacional de ADN , Exones/genética , Amplificación de Genes , Humanos , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Análisis de Secuencia de ADNRESUMEN
Gentamicin and netilmicin (internal standard) were extracted from urine using C18 solid-phase extraction cartridges (94.3% recovery) and then derivatised with o-phthalaldehyde and 3-mercaptopropionic acid. The derivative was stable for >6 h. The mobile phase methanol-glacial acetic acid-water (800:20:180, v/v), contained 0.02 M sodium heptanesulfonic acid, pH 3.4, and was passed at 1.0 ml min(-1) through a C18 column with fluorescence detection (excitation 340 nm, emission 418 nm). The four main components of gentamicin (C1, C1a, C2, C2a) and netilmicin, the internal standard, were separated. Using the C1a gentamicin peak, linearity was demonstrated from 0.5 to 10 microg ml(-1) and the limit of detection was 75 microg l(-1). Following 80-mg oral, 40-mg intravenous and 80-mg nebulised administration, the mean (SD) gentamicin urinary excretion was zero, 38.27 (0.96) and 1.93 (0.28) mg, respectively. Despite the relatively low lung deposition following inhalation of gentamicin the assay developed can be used to quantify the low urinary concentrations. Using this assay it should be possible to carry out urinary pharmacokinetic studies to identify the relative lung deposition of gentamicin following different methods of inhalation.
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Antibacterianos/orina , Cromatografía Líquida de Alta Presión/métodos , Gentamicinas/orina , o-Ftalaldehído/química , Adulto , Antibacterianos/administración & dosificación , Femenino , Gentamicinas/administración & dosificación , Humanos , Exposición por Inhalación , Espectrometría de FluorescenciaRESUMEN
The correlation ability and solubility prediction in supercritical carbon dioxide of a proposed equation were studied. The work involved the solubilities of nicotinic acid and p-acetoxyacetanilide in supercritical carbon dioxide using a dynamic flow solubility system at 35-75 degrees C and 100-200 bar. The generated experimental solubility data together with 21 data sets collected from the literature were used to evaluate the correlation ability of available empirical equations. The average absolute relative deviations (AARD) for the empirical equations are 12.6-24.8%. The prediction capability of the modified empirical relationship was studied with six experimental data points as a training set. Then, solubility at other temperatures and pressures were predicted. The AARD between predicted solubilities and observed values is 17%.
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Dióxido de Carbono/química , Acetanilidas/química , Algoritmos , Hipolipemiantes/química , Modelos Químicos , Niacina/química , Solubilidad , Solventes , TemperaturaRESUMEN
A mathematical model for calculating apparent acid dissociation constants (pK(a)) in hydroorganic mixtures with respect to the concentration of organic solvent in a binary mixture is proposed. The correlation ability of the proposed model is evaluated by employing pK(a) value of 75 different weak acids in 13 water-cosolvent systems. The results show that the equation is able to correlate the pK(a) values with an overall mean percentage differences (MPD) of 0.52+/-0.43%. In order to test the prediction capability of the model, four experimental pK(a) values for each data set have been employed to train the model, then the pK(a) values at other solvent compositions predicted and the overall MPD obtained is 1.41+/-1.15%. The applicability of the model to correlate/predict pK(a) values of structurally related drugs in a given binary solvent has been shown. The obtained overall MPD for correlation and prediction capabilities are 1.60+/-2.16 and 2.89+/-3.22%, respectively.
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Ácidos/química , Modelos Químicos , Solventes/química , 2-Propanol/química , Antiinflamatorios no Esteroideos/química , Fenómenos Químicos , Química Física , Cromatografía Líquida de Alta Presión , Interpretación Estadística de Datos , Electroforesis Capilar , Soluciones Farmacéuticas/química , Valor Predictivo de las Pruebas , Agua/químicaRESUMEN
Pigmented villonodular synovitis (PVNS) is an uncommon entity, which has the potential to cause severe pain. The gold standard for evaluation is MRI, and previous PET findings associated with PVNS have only been documented in the setting of concurrent malignancy. In the setting of recurrent disease, PET is being used to evaluate prebiological and postbiological treatment responses. Recurrent PVNS demonstrates greater hypermetabolic activity than previously documented, supporting the case as a potential mimic of malignant/metastatic disease. Post-treatment evaluations demonstrate decreased metabolic activity, which suggests response to treatment. This behaviour further supports the contention that there is a neoplastic origin to PVNS.
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Tomografía de Emisión de Positrones , Sinovitis Pigmentada Vellonodular/diagnóstico por imagen , Biopsia , Femenino , Pie , Humanos , Imagen por Resonancia Magnética , Persona de Mediana Edad , Imagen Multimodal , Sinovitis Pigmentada Vellonodular/terapia , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVES: Currently, suspensions prepared from micronised drug substances are the only delivery system marketed for nebulisation of steroids, and reported inconsistent or low bioavailability arising from their use provides a rationale for researching alternative formulations. Supercritical fluid processing of drug substances to obtain respirable-sized particles has been used over the last decade to formulate dry powder inhalers. We aimed thus to process budesonide powder to improve its deposition characteristics. METHODS: In an attempt to overcome the limitations of nebuliser suspensions when prepared from micronised drug particles, budesonide powder was processed using a supercritical fluid based process and suspended using Tween 80 as a surfactant to provide an aqueous nebuliser formulation. The in-vitro characteristics of the emitted dose on nebulisation for the prepared suspension were then compared to a commercially available suspension formulation of budesonide using a jet and a vibrating mesh nebuliser. KEY FINDINGS: The results showed a significant improvement of the in-vitro deposition properties of the suspension containing supercritical fluid engineered budesonide particles. CONCLUSIONS: The results indicated the benefit of such materials compared with traditionally micronised drug powders.