Iván Gómez-Conde

Intravital Placenta Imaging Reveals Microcirculatory Dynamics Impact on Sequestration and Phagocytosis of Plasmodium-Infected Erythrocytes

Malaria in pregnancy is exquisitely aggressive, causing a range of adverse maternal and fetal outcomes prominently linked to Plasmodium-infected erythrocyte cytoadherence to fetal trophoblast. To elucidate the physiopathology of infected erythrocytes (IE) sequestration in the placenta we devised an experimental system for intravital placental examination of P. berghei-infected mice. BALB/c females were mated to C57Bl/6 CFP+ male mice and infected with GFP+ P. berghei IE, and at gestational day 18, placentas were exposed for time-lapse imaging acquisition under two-photon microscopy. Real-time images and quantitative measurements revealed that trophoblast conformational changes transiently restrain blood flow in the mouse placental labyrinth. The complex dynamics of placental microcirculation promotes IE accumulation in maternal blood spaces with low blood flow and allows the establishment of stable IE-trophoblast contacts. Further, we show that the fate of sequestered IE includes engulfment by both macrophagic and trophoblastic fetal-derived cells. These findings reinforce the current paradigm that IE interact with the trophoblast and provide definitive evidence on two novel pathogenesis mechanisms: (1) trophoblast layer controls placental microcirculation promoting IE sequestration; and (2) fetal-derived placental cells engulf sequestered IE.

Early skin immunological disturbance after Plasmodium-infected mosquito bites.

Although the role of regulatory T cells (Tregs) during malaria infection has been studied extensively, such studies have focused exclusively on the role of Treg during the blood stage of infection; little is known about the detailed mechanisms of Tregs and sporozoite deposition in the dermis by mosquito bites. In this paper we show that sporozoites introduced into the skin by mosquito bites increase the mobility of skin Tregs and dendritic cells (DCs). We also show differences in MHC class II and/or CD86 expression on skin-resident dendritic cell subtypes and macrophages. From the observed decrease of the number of APCs into draining lymph nodes, suppression of CD28 expression in conventional CD4 T cells, and a low homeostatic proliferation of skin-migrated CD4 T found in nude mice indicate that Tregs may play a fundamental role during the initial phase of malaria parasite inoculation into the mammalian host.

A KPCA spatio-temporal differential geometric trajectory cloud classifier for recognizing human actions in a CBVR system

A CBVR based upon spatio-temporal trajectories of human motion.Trajectories encode two motion scales: large (hyperplanes); small (local differential geometry).New fuzzy KNN classifies trajectories with proximity, orientation and approximate membership.Validation of video retrieval system from standard datasets and feature length films. We describe a new algorithm for distinguishing human actions in videos, called the differential geometric trajectory cloud (DGTC) method that captures both fine and large scale structure of the covariant transformed spatio-temporal optical flow field. We show the utility of our algorithm in the context of a content based video retrieval (CBVR) system, where specific frames from a full length video (or separate video shots in a database) are identified containing a queried human action. In the DGTC method, the local geometry of the spatio-temporal covariant eigenspace curves, unique to each human action, are characterized by the Frenet-Serret basis equations, thereby specifying the local time averaged curvature and torsion, as well as providing a means for defining a mean osculating hyperplane for the entire trajectory. To classify a human action from a query, our system uses an adaptive distance metric between the covariant transformed query trajectory and each of the trajectories from all of the actions in the training set. Based upon the separation of between the query and each class, the distance uses either large or small scale information about the trajectory: for large separations, the distance is the separation between trajectory cloud centroids, while for small and intermediate separations the distance is based upon the mean hyperplane orientation obtained from the time averaged curvature and torsion of the trajectory. Our system can function in real-time and has an accuracy greater than 93% for multiple action recognition within video repositories. We also demonstrate the use of our CBVR system locating specific frame positions of trained actions in two full featured films.

Automatic recognition and scoring of olympic rhythmic gymnastic movements

We describe a conceptually simple algorithm for assigning judgement scores to rhythmic gymnastic movements, which could improve scoring objectivity and reduce judgemental bias during competitions. Our method, implemented as a real-time computer vision software, takes a video shot or a live performance video stream as input and extracts detailed velocity field information from body movements, transforming them into specialized spatio-temporal image templates. The collection of such images over time, when projected into a velocity covariance eigenspace, trace out unique but similar trajectories for a particular gymnastic movement type. By comparing separate executions of the same atomic gymnastic routine, our method assigns a quality judgement score that is related to the distance between the respective spatio-temporal trajectories. For several standard gymnastic movements, the method accurately assigns scores that are comparable to those assigned by expert judges. We also describe our rhythmic gymnastic video shot database, which we have made freely available to the human movement research community. The database can be obtained at http://www.milegroup.net/apps/gymdb/.

Tracking T and B cells from two-photon microscopy imaging using constrained SMC clusters.

This paper describes a novel software algorithm, called constrained Sequential Monte Carlo (SMC) clusters, for tracking a large collection of individual cells from intra-vital two-photon microscopy image sequences. We show how our method and software tool, implemented in python, is useful for quantifying the motility of T and B lymphocytes involved in an immune response vs lymphocytes under non immune conditions. We describe the theory behind our algorithm and briefly discuss the architecture of our software. Finally, we demonstrate both the functionality and utility of software by applying it to two practical examples from videos displaying lymphocyte motility in B cell zones (follicles) and T cell zones of lymph nodes.

Intravital microscopy technique to study parasite dynamics in the labyrinth layer of the mouse placenta.

Intravital imaging techniques are the best approach to investigate in situ cellular behavior under physiological conditions. Many techniques have emerged during these last few years for this purpose. We recently described an intravital imaging technique that allows for the observation of placenta physiological responses at the labyrinth layer of this tissue. This technique will be very useful to study many placental opportunistic infections and in this article we reinforce its usefulness by analyzing placental physiological entrapment of beads and parasites. In particular, our results show that small beads (1.0 μm) or Plasmodium chabaudi-GFP-infected-Red Blood Cells (Pc-GFP-iRBCs) cannot get trapped inside small or large blood vessels of popliteal lymph nodes (PLNs). Inside the placenta, clusters of beads could only be found inside the maternal blood vessels. However, Pc-GFP-iRBCs were found inside and outside the maternal blood vessels. We observed that trophoblasts can ingest infected-Red Blood Cells (iRBCs) in vitro and immunofluorescence of placenta revealed Pc-GFP-iRBCs inside and outside the maternal blood vessels. Taken together, we conclude that fast deposition of particles inside blood vessels seems to be an intrinsic characteristic of placenta blood flow, but iRBCs could be internalized by trophoblast cells. Thus these results represent one of the many possible uses of our intravital imaging technique to address important questions inside the parasitological field.