RESUMEN
Diagnostic electron microscopy for infectious diseases has the advantage that "everything" in the specimen can be observed, without a priori knowledge of the likely identity of the microorganisms present in the sample. The classical specimen preparation method used employs a droplet of sample, which allows particles to adsorb to a support film, and is subsequently negative stained. This "grid on drop" procedure has a sensitivity range of approximately 106 viruses per mL if no enrichment procedures are used. In the current investigation we present a novel use of filtration that allows us to detect viruses at concentrations as low as 102 viruses per mL. We present here methods based on filtration, in which total virus, and not virus concentration, is the limiting factor for detection. We show that filtration is more sensitive than conventional negative staining and can detect as few as 5 × 103 particles per sample.
Asunto(s)
Filtración/métodos , Microscopía Electrónica de Rastreo/métodos , Microscopía Electrónica de Transmisión/métodos , Animales , Bacteriófagos/ultraestructura , Línea Celular , Chlorocebus aethiops , Cricetinae , Leptospira/ultraestructura , Células Vero , Virus/ultraestructuraRESUMEN
We identified the interaction between HBV X (HBx) protein and the oncogene AIB1 (amplified in breast cancer 1). A serine/proline motif (SSPSPS) in HBx was found to be required for the interaction. Two LXD motifs [LLXX(X)L, X means any amino acids], LLRNSL and LLDQLHTLL in AIB1 were also found to be involved in the HBx-AIB1 interaction. The HBx-AIB1 interaction was important for the activation of NFκB signal transduction, the HBx mutant that did not interact with AIB1showed dramatically lower NFκB activation activity than the WT HBx. These findings contribute to the new understanding on signal transduction activation mechanisms of HBx.
Asunto(s)
Carcinógenos , FN-kappa B/metabolismo , Coactivador 3 de Receptor Nuclear/metabolismo , Transactivadores/metabolismo , Secuencia de Aminoácidos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Línea Celular Tumoral , Secuencia Conservada , Humanos , Datos de Secuencia Molecular , Mutación , Coactivador 3 de Receptor Nuclear/genética , Dominios y Motivos de Interacción de Proteínas , Serina/genética , Serina/metabolismo , Transducción de Señal , Transactivadores/genética , Técnicas del Sistema de Dos Híbridos , Proteínas Reguladoras y Accesorias ViralesRESUMEN
Homozygous sickle cell disease (SCD) presents a multitude of challenges in patients undergoing cardiac surgery with cardiopulmonary bypass. Special consideration must be made in such patients and routine practice modified to prevent hypoxia, hypothermia, acidaemia and low-flow states which may potentially trigger a fatal sickling crisis perioperatively. We discuss several perioperative management strategies including a preoperative exchange transfusion, high flow normothermic bypass and warm blood cardioplegia that was utilized in a woman with homozygous SCD who underwent a successful double valve procedure.
Asunto(s)
Anemia de Células Falciformes/complicaciones , Válvula Mitral/cirugía , Atención Perioperativa/métodos , Válvula Tricúspide/cirugía , Adulto , Anemia de Células Falciformes/fisiopatología , Puente Cardiopulmonar/métodos , Femenino , Homocigoto , Humanos , Resultado del TratamientoRESUMEN
OBJECTIVES: Sickle cell disease (SCD) is an inherited blood disorder characterized by recurrent 'crisis' pain, which is the most common reason for repeated hospital admission. The nature of this pain, however, is poorly understood, and the pain is often sub-optimally managed. METHODS: A focus group format, interpreted using thematic analysis, was used to gain a greater understanding of the barriers that SCD patients face in managing their pain and their perceptions of the treatment that they receive from healthcare professionals. RESULTS: Key issues emerging from the focus groups that adversely affected participants' pain management included: feeling isolated by their experience of 'crisis' pain, not being listened to, and limitations to social support networks. Specific issues relating to seeking medical care included lack of understanding about SCD by nonspecialist clinicians, feelings of being low priority due to the 'invisible' nature of their pain, and feeling mistrusted by medical staff when seeking analgesia. DISCUSSION: Feelings of isolation may drive maladaptive coping strategies and manifest in anger, aggression and active avoidance of service use. Suggested service improvements include the active targeting of isolated individuals in the form of pain discussion groups or self-help groups, and greater provision of specialised services.