Vesicle motion during sustained exocytosis in chromaffin cells

Chromaffin cells release catecholamines by exocytosis, a process that includes vesicle docking, priming and fusion. Although all these steps have been intensively studied, some aspects of their mechanisms, particularly those regarding vesicle transport to the active sites situated at the membrane, are still unclear. In this work, we show that it is possible to extract information on vesicle motion in Chromaffin cells from the combination of Langevin simulations and amperometric measurements. We developed a numerical model based on Langevin simulations of vesicle motion towards the cell membrane and on the statistical analysis of vesicle arrival times. We also performed amperometric experiments in bovine-adrenal Chromaffin cells under Ba2+ stimulation to capture neurotransmitter releases during sustained exocytosis. In the sustained phase, each amperometric peak can be related to a single release from a new vesicle arriving at the active site. The amperometric signal can then be mapped into a spike-series of release events. We normalized the spike-series resulting from the current peaks using a time-rescaling transformation, thus making signals coming from different cells comparable. We discuss why the obtained spike-series may contain information about the motion of all vesicles leading to release of catecholamines. We show that the release statistics in our experiments considerably deviate from Poisson processes. Moreover, the interspike-time probability is reasonably well described by two-parameter gamma distributions. In order to interpret this result we computed the vesicles’ arrival statistics from our Langevin simulations. As expected, assuming purely diffusive vesicle motion we obtain Poisson statistics. However, if we assume that all vesicles are guided toward the membrane by an attractive harmonic potential, simulations also lead to gamma distributions of the interspike-time probability, in remarkably good agreement with experiment. We also show that including the fusion-time statistics in our model does not produce any significant changes on the results. These findings indicate that the motion of the whole ensemble of vesicles towards the membrane is directed and reflected in the amperometric signals. Our results confirm the conclusions of previous imaging studies performed on single vesicles that vesicles’ motion underneath plasma membranes is not purely random, but biased towards the membrane.

Surface registration from range image fusion

The registration of full 3-D models is an important task in computer vision. Range finders only reconstruct a partial view of the object. Many authors have proposed several techniques to register 3D surfaces from multiple views in which there are basically two aspects to consider. First, poor registration in which some sort of correspondences are established. Second, accurate registration in order to obtain a better solution. A survey of the most common techniques is presented and includes experimental results of some of them

A review of sensor fusion techniques for underwater vehicle navigation

This work provides a general description of the multi sensor data fusion concept, along with a new classification of currently used sensor fusion techniques for unmanned underwater vehicles (UUV). Unlike previous proposals that focus the classification on the sensors involved in the fusion, we propose a synthetic approach that is focused on the techniques involved in the fusion and their applications in UUV navigation. We believe that our approach is better oriented towards the development of sensor fusion systems, since a sensor fusion architecture should be first of all focused on its goals and then on the fused sensors

Functional studies of the deubiquitinating enzymes USP19, USP4 and UCH-L1

El marcaje de proteínas con ubiquitina, conocido como ubiquitinación, cumple diferentes funciones que incluyen la regulación de varios procesos celulares, tales como: la degradación de proteínas por medio del proteosoma, la reparación del ADN, la señalización mediada por receptores de membrana, y la endocitosis, entre otras (1). Las moléculas de ubiquitina pueden ser removidas de sus sustratos gracias a la acción de un gran grupo de proteasas, llamadas enzimas deubiquitinizantes (DUBs) (2). Las DUBs son esenciales para la manutención de la homeostasis de la ubiquitina y para la regulación del estado de ubiquitinación de diferentes sustratos. El gran número y la diversidad de DUBs descritas refleja tanto su especificidad como su utilización para regular un amplio espectro de sustratos y vías celulares. Aunque muchas DUBs han sido estudiadas a profundidad, actualmente se desconocen los sustratos y las funciones biológicas de la mayoría de ellas. En este trabajo se investigaron las funciones de las DUBs: USP19, USP4 y UCH-L1. Utilizando varias técnicas de biología molecular y celular se encontró que: i) USP19 es regulada por las ubiquitin ligasas SIAH1 y SIAH2 ii) USP19 es importante para regular HIF-1α, un factor de transcripción clave en la respuesta celular a hipoxia, iii) USP4 interactúa con el proteosoma, iv) La quimera mCherry-UCH-L1 reproduce parcialmente los fenotipos que nuestro grupo ha descrito previamente al usar otros constructos de la misma enzima, y v) UCH-L1 promueve la internalización de la bacteria Yersinia pseudotuberculosis.

"A Sequence in Subdomain 2 of DBL1a of Plasmodium falciparum Erythrocyte Membrane Protein 1 Induces Strain Transcending Antibodies"

Immunity to severe malaria is the first level of immunity acquired to Plasmodium falciparum. Antibodies to the variant antigen PfEMP1 (P. falciparum erythrocyte membrane protein 1) present at the surface of the parasitized red blood cell (pRBC) confer protection by blocking microvascular sequestration. Here we have generated antibodies to peptide sequences of subdomain 2 of PfEMP1-DBL1a previously identified to be associated with severe or mild malaria. A set of sera generated to the amino acid sequence KLQTLTLHQVREYWWALNRKEVWKA, containing the motif ALNRKE, stained the live pRBC. 50% of parasites tested (7/14) were positive both in flow cytometry and immunofluorescence assays with live pRBCs including both laboratory strains and in vitro adapted clinical isolates. Antibodies that reacted selectively with the sequence REYWWALNRKEVWKA in a 15-mer peptide array of DBL1a-domains were also found to react with the pRBC surface. By utilizing a peptide array to map the binding properties of the elicited anti-DBL1a antibodies, the amino acids WxxNRx were found essential for antibody binding. Complementary experiments using 135 degenerate RDSM peptide sequences obtained from 93 Ugandan patient-isolates showed that antibody binding occurred when the amino acids WxLNRKE/D were present in the peptide. The data suggests that the ALNRKE sequence motif, associated with severe malaria, induces strain-transcending antibodies that react with the pRBC surface

A sequence in subdomain 2 of DBL1 alpha of plasmodium falciparum erythrocyte membrane protein 1 induces strain transcending antibodies

Immunity to severe malaria is the first level of immunity acquired to Plasmodium falciparum. Antibodies to the variant antigen PfEMP1 (P. falciparum erythrocyte membrane protein 1) present at the surface of the parasitized red blood cell (pRBC) confer protection by blocking microvascular sequestration. Here we have generated antibodies to peptide sequences of subdomain 2 of PfEMP1-DBL1 alpha previously identified to be associated with severe or mild malaria. A set of sera generated to the amino acid sequence KLQTLTLHQVREYWWALNRKEVWKA, containing the motif ALNRKE, stained the live pRBC. 50% of parasites tested (7/14) were positive both in flow cytometry and immunofluorescence assays with live pRBCs including both laboratory strains and in vitro adapted clinical isolates. Antibodies that reacted selectively with the sequence REYWWALNRKEVWKA in a 15-mer peptide array of DBL1 alpha-domains were also found to react with the pRBC surface. By utilizing a peptide array to map the binding properties of the elicited anti-DBL1 alpha antibodies, the amino acids WxxNRx were found essential for antibody binding. Complementary experiments using 135 degenerate RDSM peptide sequences obtained from 93 Ugandan patient-isolates showed that antibody binding occurred when the amino acids WxLNRKE/D were present in the peptide. The data suggests that the ALNRKE sequence motif, associated with severe malaria, induces strain-transcending antibodies that react with the pRBC surface.

Fusion 1 : science 11-14.

Proporciona una serie de recursos sobre el funcionamiento integrado del trabajo científico a los alumnos de la etapa 3 de secundaria (key stage 3). Está escrito para cumplir con los requisitos del programa de estudios de 2008, ayuda a desarrollar los conocimientos y habilidades científicos e incluye además actividades e investigaciones prácticas en cada una de las lecciones. Se acompaña del recurso para el profesor.

Fusion 1. Teacher book : science 11-14.

Ayuda a los profesores a planificar las clases de ciencias, contiene referencias al plan de estudios, objetivos y resultados del aprendizaje, habilidades funcionales, notas y sugerencias de deberes para completar las lecciones. Cumple con los requisitos del programa de estudios de 2008 y va destinado a los alumnos de la etapa 3 de secundaria (key stage 3). Se acompaña del libro del alumno.

Fusion 2 : science 11-14.

Proporciona una serie de recursos sobre el funcionamiento integrado del trabajo científico a los alumnos de la etapa 3 de secundaria (key stage 3). Está escrito para cumplir con los requisitos del programa de estudios de 2008, ayuda a desarrollar los conocimientos y habilidades científicos e incluye además actividades e investigaciones prácticas en cada una de las lecciones. Se acompaña del recurso para el profesor.

Fusion 2. Teacher book : science 11-14.

Ayuda a los profesores a planificar las clases de ciencias, contiene referencias al plan de estudios, objetivos y resultados del aprendizaje, habilidades funcionales, notas y sugerencias de deberes para completar las lecciones. Cumple con los requisitos del programa de estudios de 2008 y va destinado a los alumnos de la etapa 3 de secundaria (key stage 3). Se acompaña del libro del alumno.

Fusion 3 : science 11-14.

Proporciona una serie de recursos sobre el funcionamiento integrado del trabajo científico a los alumnos de la etapa 3 de secundaria (key stage 3). Está escrito para cumplir con los requisitos del programa de estudios de 2008, ayuda a desarrollar los conocimientos y habilidades científicos e incluye además actividades e investigaciones prácticas en cada una de las lecciones. Se acompaña del recurso para el profesor.

Fusion 3. Teacher book : science 11-14.

Ayuda a los profesores a planificar las clases de ciencias, contiene referencias al plan de estudios, objetivos y resultados del aprendizaje, habilidades funcionales, notas y sugerencias de deberes para completar las lecciones. Cumple con los requisitos del programa de estudios de 2008 y va destinado a los alumnos de la etapa 3 de secundaria (key stage 3). Se acompaña del libro del alumno.

Development of a decision support system for the integrated control of membrane bioreactors

The work presented in this PhD thesis includes various partial studies aimed at developing a decision support system for membrane bioreactor integrated control. The decision support systems (DSS) have as a main goal to facilitate the operation of complex processes due to the multiple variables that are processed. For this reason, the research used has focused on aspects related to nutrient removal, and on the development of indicators or sensors capable of facilitating, automating and controlling the filtration process in an integrated way with the biological processes that taking place. Work has also been done on the design, development, implementation and validation of tools based on the knowledge made available by the automatic control and the supervision of the MBRs

Development of an air-scour control system for membrane bioreactors

The thesis involves the development and implementation of a new and robust control system based on permeability trends but at the same time capable of reducing aeration proportionally to permeate flux. Permeability was made a key parameter for directly comparing temporary changes in membrane performance. Transmembrane pressure and flux were gathered every 10 seconds and permeability values were automatically calculated; different mathematical algorithms were applied for the signal filtering of on-line data. Short term and long term permeability trends were compared once a day, and a control action was applied proportionally to the short term/long term permeability ratio without exceeding the aeration flow recommended by the membrane suppliers.

The effects of extracorporeal membrane oxygenation (ECMO) on auditory brainstem responses (ABR) in neonates

This paper presents a study of the effects of extracorporeal membrane oxygenation (ECMO) on the auditory brainstem response (ABR) of twenty-three neonates, and whether there was asymmetric ABRs in the neonates who had ECMO.