Do we have today a reliable method to detect the moment of loss of consciousness during induction of general anaesthesia? / ¿Contamos actualmente con un método fiable para detectar el momento de pérdida de consciencia durante la inducción de la anestesia?

This review aims to give an overview of the current state of monitoring depth of anaesthesia and detecting the moment of loss of consciousness, from the first clinical signs involved in anaesthesia to the latest technologies used in this area. Such techniques are extremely important for the development of automatic systems for anaesthesia control, including preventing intraoperative awareness episodes and overdoses. A search in the databases Pubmed and IEEE Xplore was performed using terms such anaesthetic monitoring, depth of anaesthesia, loss of consciousness, as well as anaesthesia indexes, namely BIS. Despite the several methods capable of monitoring the hypnotic state of anaesthesia, there is still no methodology to accurate detect the moment of loss of consciousness during induction of general anaesthesia.

Acquisition and analysis of 3D mandibular movement using a device based on electromagnetic sensors and a neural network.

In dental medicine, the study of mandibular movement has an important role in oral rehabilitation as it allows diagnosis of pathologies in the temporomandibular joint and the definition of adequate treatment plans. In this paper, a new device specially developed for the acquisition and analysis of 3D mandibular movement is presented. A facial arc is adopted as its main support structure and electromagnetic sensors are employed to acquire the mandibular movement. A neural network is used to transform the electrical signals output by the sensors into 3D Cartesian coordinates. A custom-made computer application is developed to display and analyse the movement acquired. The device is shown to be easy to use, comfortable for patients and capable of being produced at an affordable price.

Inhibition of interferon-gamma-induced nitric oxide production in endotoxin-activated macrophages by cytolethal distending toxin.

INTRODUCTION: Cytolethal distending toxin (CDT) is a DNA-targeting agent produced by certain pathogenic gram-negative bacteria such as the periodontopathogenic organism Aggregatibacter actinomycetemcomitans. CDT targets lymphocytes and other cells causing cell cycle arrest and apoptosis, impairing the host immune response and contributing to the persistence of infections caused by this microorganism. In this study we explored the effects of CDT on the innate immune response, by investigating how it affects production of nitric oxide (NO) by macrophages. METHODS: Murine peritoneal macrophages were stimulated with Escherichia coli sonicates and NO production was measured in the presence or not of active CDT. RESULTS: We observed that CDT promptly and significantly inhibited NO production by inducible nitric oxide synthase (iNOS) in a dose-dependent manner. This inhibition is directed towards interferon-gamma-dependent pathways and is not mediated by either interleukin-4 or interleukin-10. CONCLUSION: This mechanism may constitute an important aspect of the immunosuppression mediated by CDT and may have potential clinical implications in A. actinomycetemcomitans infections.

Cloning of a thymic stromal cell capable of protecting thymocytes from apoptosis.

A clone of thymic stromal cells, namely 2BH4, was established by primary culture, cellular transfection and limiting dilution. Morphological analysis by transmission electron microscopy revealed that these cells grow as multilayers, producing a well-defined basement membrane to which they attach and frequently form structures similar to hemidesmosomes. The adjoining cells are connected by intercellular junctions, as tight junctions, intermediate junctions, and desmosome-like junctions, as well as interdigitations. Their cytoplasm contains microtubules, strands of actin filaments, and scarce intermediate filaments. Fluorescence microscopy revealed that 2BH4 cells stain with anti-cytokeratin antibodies, the majority of them giving a faint reaction. In addition, they express Thy-1.1, LFA-1, ICAM-1, and the gp23 epithelial antigen, and synthesize laminin. They have a doubling time of 16 hr and are able to bind thymocytes. Thymocytes cultured in the presence of 2BH4 cells are partially protected from both spontaneous and PMA- or dexamethasone-induced apoptosis. This protection is conferred neither by soluble factors normally produced by the 2BH4 cells nor by the sole contact with fixed 2BH4 cells. Rather, thymocytes must interact with metabolically active 2BH4 cells in order to receive the protective signal(s).

Colorimetric assay for the measurement of thymocyte/thymic stromal cell adhesion.

1. We describe a simple and accurate colorimetric adhesion assay (CAA) and illustrate the assay by measuring the adhesion of mouse thymocytes to mouse 2BH4 cells. 2. The assay is based on the crystal violet staining of thymocytes adhered to a subconfluent layer of 2BH4 cells (plated at 2 x 10(4) cells/well for 24 h). The optimal incubation time was shown to be 1 h and washing in PBS of non-bound and non-specifically bound thymocytes is the critical step for the precision and accuracy of the assay. 3. Saturation curves were obtained for thymocytes adhered to plated 2BH4 cells. The blank (only 2BH4 cells) was near 0.200 +/- 0.010 (mean +/- SD) and was quite reproducible. As expected, the extent of adhesion was also dependent on the number of plated 2BH4 cells. Standard curves need to be run with each assay for quantitative measurements. The intra-assay and interassay coefficients of variation were 5% and 20%, respectively. 4. The specificity of the reaction was demonstrated by the reduction of adherence by trypsin pretreatment of thymocytes, and the dose-dependent inhibition of adherence by rabbit anti-mouse thymocyte antisera but not rabbit anti-mouse immunoglobulin antisera. 5. The proposed method is simple and requires less effort than the counting of adhered cells with the light microscope and does not require the use of radioactive material as when labelling with Na2(51)CrO4 is utilized.

Colorimetric assay for the measurement of thymocyte/thymic stromal cell adhesion

1. We describe a simple and accurate colorimetric adhesion assay (CAA) and illustrate the assay by measuring the adhesion of mouse thymocytes to mouse 2BH4 cells. 2. The assay is based on the crystal violet staining of thymocytes adhered to a subconfluent layer of 2BH4 cells (plated at 2 x 10(4) cells/well for 24 h). The optimal incubation time was shown to be 1 h and washing in PBS of non-bound and non-specifically bound thymocytes is the critical step for the precision and accuracy of the assay. 3. Saturation curves were obtained for thymocytes adhered to plated 2BH4 cells. The blank (only 2BH4 cells) was near 0.200 +/- 0.010 (mean +/- SD) and was quite reproducible. As expected, the extent of adhesion was also dependent on the number of plated 2BH4 cells. Standard curves need to be run with each assay for quantitative measurements. The intra-assay and interassay coefficients of variation were 5 and 20, respectively. 4. The specificity of the reaction was demonstrated by the reduction of adherence by trypsin pretreatment of thymocytes, and the dose-dependent inhibition of adherence by rabbit anti-mouse thymocyte antisera but not rabbit anti-mouse immunoglobulin antisera. 5. The proposed method is simple and requires less effort than the counting of adhered cells with the light microscope and does not require the use of radioactive material as when labelling with Na2(51)CrO4 is utilized.

Identification of a 16-kDa thymocyte membrane glycoprotein involved in the thymocyte/thymic medullary epithelial cell interaction.

We have previously described a type of lymphoepithelial interaction involving CD4+ CD8+ thymocytes and a medullary epithelial cell line (E-5). This interaction is mediated by the recently described gp23/45 epithelial adhesion molecule and an as yet unknown thymocyte receptor. The present work describes a thymocyte surface glycoprotein of 16 kDa which binds both to E-5 cells and to the purified gp23/45 adhesion molecule. In addition, a thymic lymphoma cell line (Ti-6), which interacts with the E-5 cells via the gp23/45 receptor, also present a 16-kDa glycoprotein on its surface. Taken together, the data suggest that the 16-kDa thymocyte surface glycoprotein participates in the binding between these cells and the thymic epithelium.

Suppression of antibody response to an unrelated antigen in experimental murine paracoccidioidomycosis: effect of cyclophosphamide and indomethacin.

A suppressive effect of experimental paracoccidioidomycosis on the IgE antibody response to an unrelated antigen (ovalbumin) has been previously observed in mice. This effect was restricted to a short period, reaching maximum levels when OA was administered on the third day of Pb-infection. In order to study possible mechanisms involved in the establishment of this suppression, resistant (A/SN) and susceptible (B10.A) mice were treated with either a low dose of cyclophosphamide (CY) or indomethacin (INDO), a potent inhibitor of prostaglandin synthesis. While treatment with the first drug in A/SN mice induced only a recovery of the IgE anti-OA antibody response, in B10.A mice this effect was extended to IgG1, IgG2a and total levels of anti-OA antibodies. On the other hand, treatment with INDO reverted the anti-OA antibody suppression regarding each antibody class tested and in both strains of mice. These results suggest the participation of prostaglandins and of a cyclophosphamide-sensitive mechanism in the induction of the suppressive phenomenon in the human system.

Suppression of IgE antibody production against an unrelated antigen in experimental murine paracoccidioidomycosis.

A suppression of the IgE antibody response to ovalbumin was obtained in susceptible mice infected with Paracoccidioides brasiliensis yeast cells a few days prior to immunization with the former antigen plus adjuvant. A direct relationship between the number of injected fungi and the suppressive effect was established. When infection with a pathogenic isolate of P. brasiliensis (Pb 18) was compared to a non-pathogenic isolate (IVIC Pb267), the IgE anti-ovalbumin response was reduced by both. A similar effect was observed if mice were injected with dead yeast cells prior to immunization. Two strains of mice with completely opposite susceptibilities to infection with Pb18 cells (B10.A--susceptible and A/SN--resistant) both showed suppressed IgE anti-ovalbumin antibody production when infected 3 days prior to immunization. Injection of both strains of mice with P. brasiliensis antigen on the same day as immunization also had the same suppressive effect. These results suggest that the suppression of IgE response to an unrelated antigen in experimental murine paracoccidioidomycosis could be due to antigenic competition or to a suppressive component present in P. brasiliensis cells.