RESUMEN
CD30 is expressed on a variety of B-cell lymphomas, such as Hodgkin lymphoma, primary effusion lymphoma, and a diffuse large B-cell lymphoma subgroup. In normal tissues, CD30 is expressed on some activated B and T lymphocytes. However, the physiological function of CD30 signaling and its contribution to the generation of CD30+ lymphomas are still poorly understood. To gain a better understanding of CD30 signaling in B cells, we studied the expression of CD30 in different murine B-cell populations. We show that B1 cells expressed higher levels of CD30 than B2 cells and that CD30 was upregulated in IRF4+ plasmablasts (PBs). Furthermore, we generated and analyzed mice expressing a constitutively active CD30 receptor in B lymphocytes. These mice displayed an increase in B1 cells in the peritoneal cavity (PerC) and secondary lymphoid organs as well as increased numbers of plasma cells (PCs). TI-2 immunization resulted in a further expansion of B1 cells and PCs. We provide evidence that the expanded B1 population in the spleen included a fraction of PBs. CD30 signals seemed to enhance PC differentiation by increasing activation of NF-κB and promoting higher levels of phosphorylated STAT3 and STAT6 and nuclear IRF4. In addition, chronic CD30 signaling led to B-cell lymphomagenesis in aged mice. These lymphomas were localized in the spleen and PerC and had a B1-like/plasmablastic phenotype. We conclude that our mouse model mirrors chronic B-cell activation with increased numbers of CD30+ lymphocytes and provides experimental proof that chronic CD30 signaling increases the risk of B-cell lymphomagenesis.
Asunto(s)
Linfocitos B/inmunología , Linfocitos B/patología , Transformación Celular Neoplásica/patología , Antígeno Ki-1/inmunología , Linfoma de Células B/metabolismo , Animales , Antígeno Ki-1/metabolismo , Linfoma de Células B/inmunología , Linfoma de Células B/patología , Ratones , Ratones Transgénicos , Células Plasmáticas/metabolismo , Células Plasmáticas/patología , Células Precursoras de Linfocitos B/metabolismo , Células Precursoras de Linfocitos B/patología , Transducción de Señal/fisiologíaRESUMEN
BACKGROUND: Since June 1st, 2014 the use of cage beds, a physical coercive measure in psychiatric wards, is forbidden by Austrian law. The 3rd Psychiatric Department at the Otto Wagner Hospital in Vienna carried out a pilot-study prior to the application of this law and compared 6 months with and 6 months without the use of cage beds in psychiatric acute treatment. The investigation focused on the use of mechanical restraints, especially of 4-point-restraints at admission time. METHODS: The study collected clinical and epidemiological data from the patients and analyzed interviews taken with patients and staff-members after the use of coercive measures. RESULTS: The prohibition of cage beds was associated with a relative increase of the use of 4-point-restraints. Time spent in 4-point-restraints is less than time spent in the cage beds. A trend could also be observed by a decrease of time spent in 4-point-restraints. Data from the interviews showed that there was no preference by patients of either kind of physical restraint. They strongly urged for more personal talks and non-physical and non-pharmacological therapies during admission. CONCLUSIONS: Team members required more human resources for the treatment of these acute psychiatric patients.
Asunto(s)
Lechos , Coerción , Trastornos Mentales/epidemiología , Trastornos Mentales/terapia , Servicio de Psiquiatría en Hospital/legislación & jurisprudencia , Servicio de Psiquiatría en Hospital/estadística & datos numéricos , Restricción Física/legislación & jurisprudencia , Restricción Física/estadística & datos numéricos , Adulto , Austria , Femenino , Humanos , Entrevista Psicológica , Masculino , Trastornos Mentales/psicología , Persona de Mediana Edad , Admisión del Paciente , Satisfacción del Paciente , Proyectos Piloto , Revisión de Utilización de RecursosRESUMEN
A novel approach for the selective extraction of organic target compounds from water samples has been developed using a mixed-bed solid phase extraction (mixed-bed SPE) technique. The molecularly imprinted polymer (MIP) particles are embedded in a network of silica gel to form a stable uniform porous bed. The capabilities of this method are demonstrated using atrazine as a model compound. In comparison to conventional molecularly imprinted-solid phase extraction (MISPE), the proposed mixed-bed MISPE method in combination with gas chromatography-mass spectrometry (GC-MS) analysis enables more reproducible and efficient extraction performance. After optimization of operational parameters (polymerization conditions, bed matrix ingredients, polymer to silica gel ratio, pH of the sample solution, breakthrough volume plus washing and elution conditions), improved LODs (1.34 µg L(-1) in comparison to 2.25 µg L(-1) obtained using MISPE) and limits of quantification (4.5 µg L(-1) for mixed-bed MISPE and 7.5 µg L(-1) for MISPE) were observed for the analysis of atrazine. Furthermore, the relative standard deviations (RSDs) for atrazine at concentrations between 5 and 200 µg L(-1) ranged between 1.8% and 6.3% compared to MISPE (3.5-12.1%). Additionally, the column-to-column reproducibility for the mixed-bed MISPE was significantly improved to 16.1%, compared with 53% that was observed for MISPE. Due to the reduced bed-mass sorbent and at optimized conditions, the total amount of organic solvents required for conditioning, washing and elution steps reduced from more than 25 mL for conventional MISPE to less than 2 mL for mixed-bed MISPE. Besides reduced organic solvent consumption, total sample preparation time of the mixed-bed MISPE method relative to the conventional MISPE was reduced from more than 20 min to less than 10 min. The amount of organic solvent required for complete elution diminished from 3 mL (conventional MISPE) to less than 0.4 mL with the mixed-bed technique shows its inherent potential for online operation with an analytical instrument. In order to evaluate the selectivity and matrix effects of the developed mixed-bed MISPE method, it was applied as an extraction technique for atrazine from environmental wastewater and river water samples.
Asunto(s)
Atrazina/análisis , Herbicidas/análisis , Polímeros/química , Extracción en Fase Sólida/métodos , Contaminantes Químicos del Agua/análisis , Cromatografía de Gases y Espectrometría de Masas , Geles/química , Concentración de Iones de Hidrógeno , Impresión Molecular , Compuestos Orgánicos/química , Reproducibilidad de los Resultados , Dióxido de Silicio/química , Solventes/química , Estireno/química , Temperatura , Agua/químicaRESUMEN
INTRODUCTION: Staff time is a relevant resource in the delivery of health care interventions. Its measurement is a prerequisite for unit costing but usually complex. The aim of this study was to analyse the distribution of surgeons' work time among types and places of activities. A second aim was to use these data to calculate costs per unit of output. METHODS: A self-reporting work sampling study was carried out at a department of Urology. All of twelve surgeons involved in clinical care participated in a two-week analysis of their work time. RESULTS: A total of 2,485 data-points were collected, representing about 1,242 hours of work time. Surgeons spent the greater part of their work time in direct patient care, but substantial shares were required for documentation and organisation. Assistants were mainly required at the wards and consultants at the operating theatre and the outpatient unit. Staff costs of surgeons were 32 and 29 per patient day at the wards, respectively, 1.30 per minute at the operating theatre and 32 per visit at the outpatient unit. CONCLUSION: Results provided a basis for costing of health care interventions at the study site. However, future research should focus on the establishment of standardised terminology in order to increase transferability of results.
Asunto(s)
Procedimientos Quirúrgicos Urológicos , Servicio de Urología en Hospital , Urología , Carga de Trabajo , Femenino , Humanos , MasculinoRESUMEN
Phytoscreening of phytoremediation-based plantings is discussed as a promising monitoring tool in literature. We developed and applied an analytical procedure for the in vivo sampling of methyl tert-butyl ether (MTBE) in the common reed (Phragmites australis) from a phytoremediation site highly polluted with MTBE. The approach uses solid-phase microextraction (SPME) with the SPME fibre directly introduced into the aerenchyma of the plant stem. For optimising the analytical procedure and estimating the capability of the proposed method, laboratory tests on the microcosm scale and field studies over one vegetation period were carried out. Furthermore, the results of in vivo SPME sampling were compared with those obtained with the traditional approach for analysing plants using dynamic headspace analysis. The MTBE signals detected within the plants were also correlated with the concentration in the water phase. The discussion of results showed the feasibility of the proposed method for a qualitative phytoscreening of volatile organic compounds present in wetland plants.
Asunto(s)
Éteres Metílicos/análisis , Poaceae/química , Microextracción en Fase Sólida/métodos , Compuestos Orgánicos Volátiles/análisis , Contaminantes Químicos del Agua/análisis , Biodegradación Ambiental , Monitoreo del Ambiente/métodos , HumedalesRESUMEN
In order to investigate the fate of volatile pollutants such as methyl tert-butyl ether (MTBE) and benzene during the treatment of contaminated water using constructed wetlands, appropriate analytical methods for the analysis of wetland marsh plants, in our case common reed (Phragmites australis), are required. Different sampling procedures and different headspace (HS) based sample preparation techniques were examined to select and establish the most suited procedure for determining the target analytes in plant material. Static HS, dynamic HS and HS solid phase microextraction (SPME) in combination with GC-MS were optimized and evaluated regarding the extraction yields and their capability for quantitative analysis. Only dynamic HS analysis at 80 degrees C for 45min with trapping the analytes on Tenax TA/Chromosorb-106 desorption tubes and the subsequent thermodesorption GC-MS permits the quantitative analysis of MTBE and benzene in reed in a concentration range from 4ng up to 4mug per sample weight (approximately 1g). Static HS and HS SPME analyses were found to be less reliable due to the lack of suitable reference materials. Therefore, these methods do not permit the accurate quantification of pollutant content. Additionally, the HS SPME method is characterized by a restricted linear range of calibration curves. The optimized dynamic HS method was successfully applied for the quantitative analysis of MTBE and benzene within the plants. Their distribution within the plant depending on its height shows a different behavior due to differences in degradability of both substances. While a strong decrease of the concentration of benzene with increasing height of plant was found, the decrease of the concentration of MTBE was not as obvious as observed for benzene. Furthermore, the assessment of plant uptake during phytoremediation was demonstrated by analyzing complete plants for the constructed wetlands investigated.
Asunto(s)
Benceno/análisis , Éteres Metílicos/análisis , Poaceae/metabolismo , Contaminantes Químicos del Agua/análisis , Humedales , Cromatografía de Gases y Espectrometría de Masas , Microextracción en Fase SólidaRESUMEN
Quidquid agis, prudenter agas et respice finem!-Whatever you do, do it wisely and consider the goal. In consideration of that sage advice, the chicken B-cell line DT40 is an excellent model cell system to study the function of vertebrate genes. In addition to being highly amenable to gene manipulations, the recent influx of genome and gene/protein resources allows for the straightforward selection, design, and targeting of candidate genes for knockout analysis. This chapter will give a step by step standardized protocol to creating a gene knockout mutant in DT40. With careful consideration, the methods and protocols described herein can be easily modified to allow for further gene manipulations such as creating a knockin or a conditional mutant.
Asunto(s)
Linfocitos B/metabolismo , Pollos/genética , Animales , Secuencia de Bases , Bolsa de Fabricio/citología , Bolsa de Fabricio/metabolismo , Línea Celular , Secuencia Conservada , Cartilla de ADN/genética , ADN Complementario/genética , Etiquetas de Secuencia Expresada , Eliminación de Gen , Técnicas Genéticas , Mutación , Fenotipo , TransfecciónRESUMEN
Ig hypermutation is limited to a region of approximately 2 kb downstream of the transcription start sites of the Ig loci. The process requires transcription and the presence of Ig enhancer sequences, and is initiated by the activation-induced cytidine deaminase (AID)-mediated deamination of cytidine bases. It remains unknown why AID causes mutations selectively in the Ig genes and not in most other transcribed loci of B cells. In this study, we report that the inactivation of the E2A gene strongly reduces the rate of Ig L chain mutations in the chicken B cell line DT40 without affecting the levels of surface Ig or AID expression. The defect is complemented by the expression of cDNAs corresponding to either of the two E2A splice variants E12 or E47. The results suggest that E2A-encoded proteins enhance Ig hypermutation by recruitment of AID to the Ig loci.
Asunto(s)
Proteínas Aviares/biosíntesis , Proteínas Aviares/genética , Hipermutación Somática de Inmunoglobulina , Factores de Transcripción TCF/biosíntesis , Factores de Transcripción TCF/genética , Animales , Proteínas Aviares/deficiencia , Proteínas Aviares/fisiología , Secuencia de Bases , Línea Celular , Pollos , Células Clonales , Citidina Desaminasa/biosíntesis , Citidina Desaminasa/genética , Citidina Desaminasa/metabolismo , ADN Complementario/biosíntesis , Silenciador del Gen , Prueba de Complementación Genética , Marcadores Genéticos , Cadenas Ligeras de Inmunoglobulina/genética , Datos de Secuencia Molecular , Receptores de Antígenos de Linfocitos B/deficiencia , Receptores de Antígenos de Linfocitos B/genética , Análisis de Secuencia de ADN , Factores de Transcripción TCF/deficiencia , Factores de Transcripción TCF/fisiologíaRESUMEN
BACKGROUND: Neuropsychological studies have provided evidence for deficits in psychiatric disorders, such as schizophrenia and mood disorders. However, neuropsychological function in Panic Disorder (PD) or PD with a comorbid diagnosis of Major Depressive Disorder (MDD) has not been comprehensively studied. The present study investigated neuropsychological functioning in patients with PD and PD + MDD by focusing on tasks that assess attention, psychomotor speed, executive function, decision-making, and affective processing. METHODS: Twenty-two unmedicated patients with PD, eleven of whom had a secondary diagnosis of MDD, were compared to twenty-two healthy controls, matched for gender, age, and intelligence on tasks of attention, memory, psychomotor speed, executive function, decision-making, and affective processing from the Cambridge Neuropsychological Test Automated Battery (CANTAB), Cambridge Gamble Task, and Affective Go/No-go Task. RESULTS: Relative to matched healthy controls, patients with PD + MDD displayed an attentional bias toward negatively-valenced verbal stimuli (Affective Go/No-go Task) and longer decision-making latencies (Cambridge Gamble Task). Furthermore, the PD + MDD group committed more errors on a task of memory and visual discrimination compared to their controls. In contrast, no group differences were found for PD patients relative to matched control subjects. LIMITATIONS: The sample size was limited, however, all patients were drug-free at the time of testing. CONCLUSIONS: The PD + MDD patients demonstrated deficits on a task involving visual discrimination and working memory, and an attentional bias towards negatively-valenced stimuli. In addition, patients with comorbid depression provided qualitatively different responses in the areas of affective and decision-making processes.
Asunto(s)
Toma de Decisiones , Trastorno Depresivo Mayor/psicología , Trastorno de Pánico/psicología , Psicofisiología , Adulto , Trastornos del Conocimiento/complicaciones , Trastorno Depresivo Mayor/complicaciones , Femenino , Humanos , Masculino , Pruebas Neuropsicológicas , Trastorno de Pánico/complicacionesRESUMEN
A large number of cDNA inserts were sequenced from a high-quality library of chicken bursal lymphocyte cDNAs. Comparisons to public gene databases indicate that the cDNA collection represents more than 2,000 new, full-length transcripts. This resource defines the structure and the coding potential of a large fraction of B-cell specific and housekeeping genes whose function can be analyzed by disruption in the chicken DT40 B-cell line.
Asunto(s)
Proteínas Aviares/genética , Proteínas Aviares/metabolismo , Bolsa de Fabricio/citología , Pollos/genética , Biología Computacional , ADN Complementario/genética , Linfocitos/metabolismo , Animales , Proteínas Aviares/química , Composición de Base/genética , Bolsa de Fabricio/metabolismo , Línea Celular , Clonación Molecular , Codón Iniciador/genética , Secuencia Conservada , Bases de Datos Genéticas , Especificidad de Órganos , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Análisis de Secuencia de ADN , Homología de Secuencia , Transcripción Genética/genéticaRESUMEN
BACKGROUND: Neuropsychological deficits have been reported in patients with major depressive disorder (MDD) during an acute episode of MDD. Little is known whether these abnormalities persist when patients are remitted. The purpose of the present study was to describe the neuropsychological functioning of fully remitted, unmedicated patients with a history of MDD by focusing on tasks related to prefrontal cortex functioning. METHODS: Twenty-eight young to middle-aged, unmedicated, fully remitted patients with MDD were compared to 23 healthy control subjects on tasks from the Cambridge Automated Neuropsychological Test Battery (CANTAB) and the California Verbal Learning Test (CVLT). RESULTS: Patients with remitted MDD relative to controls were impaired on tasks of rapid visual information processing (RVIP), psychomotor performance and spatial working memory (SWM). After correction for residual depressive symptoms, deficits in sustained attention remained significant. LIMITATIONS: CANTAB tasks are not equated for difficulty, and difficulty differences between the CANTAB tasks and the CVLT are not known. CONCLUSIONS: These findings suggest deficits in sustained attention as vulnerability marker for MDD. The functional importance of this finding and the neuronal networks involved remain to be elucidated.