Miguel Lopez

Social motility in african trypanosomes.

African trypanosomes are devastating human and animal pathogens that cause significant human mortality and limit economic development in sub-Saharan Africa. Studies of trypanosome biology generally consider these protozoan parasites as individual cells in suspension cultures or in animal models of infection. Here we report that the procyclic form of the African trypanosome Trypanosoma brucei engages in social behavior when cultivated on semisolid agarose surfaces. This behavior is characterized by trypanosomes assembling into multicellular communities that engage in polarized migrations across the agarose surface and cooperate to divert their movements in response to external signals. These cooperative movements are flagellum-mediated, since they do not occur in trypanin knockdown parasites that lack normal flagellum motility. We term this behavior social motility based on features shared with social motility and other types of surface-induced social behavior in bacteria. Social motility represents a novel and unexpected aspect of trypanosome biology and offers new paradigms for considering host-parasite interactions.

Propulsion of African trypanosomes is driven by bihelical waves with alternating chirality separated by kinks.

Trypanosoma brucei, a parasitic protist with a single flagellum, is the causative agent of African sleeping sickness. Propulsion of T. brucei was long believed to be by a drill-like, helical motion. Using millisecond differential interference-contrast microscopy and analyzing image sequences of cultured procyclic-form and bloodstream-form parasites, as well as bloodstream-form cells in infected mouse blood, we find that, instead, motility of T. brucei is by the propagation of kinks, separating left-handed and right-handed helical waves. Kink-driven motility, previously encountered in prokaryotes, permits T. brucei a helical propagation mechanism while avoiding the large viscous drag associated with a net rotation of the broad end of its tapering body. Our study demonstrates that millisecond differential interference-contrast microscopy can be a useful tool for uncovering important short-time features of microorganism locomotion.

Predictive Torque Control of a Multidrive System Fed by a Dual Indirect Matrix Converter

This paper presents an experimental implementation of a predictive control strategy for a system made up of two induction machines fed by a six-leg indirect matrix converter. The objective is to evaluate the practical feasibility of this method, considering the large computational cost, and to incorporate available market technologies. This strategy achieves tracking of torque, speed, and flux references on both machines, minimizing the instantaneous input reactive power present in the system. A theoretical explanation of the main concepts that are used in the predictive control strategy is presented, along with comments on the experimental implementation, experimental setup, and results obtained under both balanced and unbalanced grid conditions.

Insect Stage-Specific Adenylate Cyclases Regulate Social Motility in African Trypanosomes

ABSTRACT Sophisticated systems for cell-cell communication enable unicellular microbes to act as multicellular entities capable of group-level behaviors that are not evident in individuals. These group behaviors influence microbe physiology, and the underlying signaling pathways are considered potential drug targets in microbial pathogens. Trypanosoma brucei is a protozoan parasite that causes substantial human suffering and economic hardship in some of the most impoverished regions of the world. T. brucei lives on host tissue surfaces during transmission through its tsetse fly vector, and cultivation on surfaces causes the parasites to assemble into multicellular communities in which individual cells coordinate their movements in response to external signals. This behavior is termed “social motility,” based on its similarities with surface-induced social motility in bacteria, and it demonstrates that trypanosomes are capable of group-level behavior. Mechanisms governing T. brucei social motility are unknown. Here we report that a subset of receptor-type adenylate cyclases (ACs) in the trypanosome flagellum regulate social motility. RNA interference-mediated knockdown of adenylate cyclase 6 (AC6), or dual knockdown of AC1 and AC2, causes a hypersocial phenotype but has no discernible effect on individual cells in suspension culture. Mutation of the AC6 catalytic domain phenocopies AC6 knockdown, demonstrating that loss of adenylate cyclase activity is responsible for the phenotype. Notably, knockdown of other ACs did not affect social motility, indicating segregation of AC functions. These studies reveal interesting parallels in systems that control social behavior in trypanosomes and bacteria and provide insight into a feature of parasite biology that may be exploited for novel intervention strategies.

Insect Stage-Specific Receptor Adenylate Cyclases Are Localized to Distinct Subdomains of the Trypanosoma brucei Flagellar Membrane

ABSTRACT Increasing evidence indicates that the Trypanosoma brucei flagellum (synonymous with cilium) plays important roles in host-parasite interactions. Several studies have identified virulence factors and signaling proteins in the flagellar membrane of bloodstream-stage T. brucei, but less is known about flagellar membrane proteins in procyclic, insect-stage parasites. Here we report on the identification of several receptor-type flagellar adenylate cyclases (ACs) that are specifically upregulated in procyclic T. brucei parasites. Identification of insect stage-specific ACs is novel, as previously studied ACs were constitutively expressed or confined to bloodstream-stage parasites. We show that procyclic stage-specific ACs are glycosylated, surface-exposed proteins that dimerize and possess catalytic activity. We used gene-specific tags to examine the distribution of individual AC isoforms. All ACs examined localized to the flagellum. Notably, however, while some ACs were distributed along the length of the flagellum, others specifically localized to the flagellum tip. These are the first transmembrane domain proteins to be localized specifically at the flagellum tip in T. brucei, emphasizing that the flagellum membrane is organized into specific subdomains. Deletion analysis reveals that C-terminal sequences are critical for targeting ACs to the flagellum, and sequence comparisons suggest that differential subflagellar localization might be specified by isoform-specific C termini. Our combined results suggest insect stage-specific roles for a subset of flagellar adenylate cyclases and support a microdomain model for flagellar cyclic AMP (cAMP) signaling in T. brucei. In this model, cAMP production is compartmentalized through differential localization of individual ACs, thereby allowing diverse cellular responses to be controlled by a common signaling molecule.

Approaches for Functional Analysis of Flagellar Proteins in African Trypanosomes

The eukaryotic flagellum is a highly conserved organelle serving motility, sensory, and transport functions. Although genetic, genomic, and proteomic studies have led to the identification of hundreds of flagellar and putative flagellar proteins, precisely how these proteins function individually and collectively to drive flagellum motility and other functions remains to be determined. In this chapter we provide an overview of tools and approaches available for studying flagellum protein function in the protozoan parasite Trypanosoma brucei. We begin by outlining techniques for in vitro cultivation of both T. brucei life cycle stages, as well as transfection protocols for the delivery of DNA constructs. We then describe specific assays used to assess flagellum function including flagellum preparation and quantitative motility assays. We conclude the chapter with a description of molecular genetic approaches for manipulating gene function. In summary, the availability of potent molecular tools, as well as the health and economic relevance of T. brucei as a pathogen, combine to make the parasite an attractive and integral experimental system for the functional analysis of flagellar proteins.

A Simple Current Control Strategy for a Four-Leg Indirect Matrix Converter

In this paper, the experimental validation of a predictive current control strategy for a four-leg indirect matrix converter is presented. The four-leg indirect matrix converter can supply energy to an unbalanced three-phase load while providing a path for the zero sequence load. The predictive current control technique is based on the optimal selection among the valid switching states of the converter by evaluating a cost function, resulting in a simple approach without the necessity for modulators. Furthermore, zero dc-link current commutation is achieved by synchronizing the state changes in the input stage with the application of a zero-voltage space vector in the inverter stage. Simulation results are presented and the strategy is experimentally validated using a laboratory prototype.

A stochastic economic dispatch model with renewable energies considering demand and generation uncertainties

This paper proposes a methodology to model and solve the problem of stochastic economic dispatch incorporating renewable energies. In this context, demand and generation randomness (wind speed, solar radiation and rates of failure) are considered. Demand, wind speed, solar radiation and unavailability are modeled through Normal, Weibull, Beta and Uniform distributions respectively. The problem is therefore recognized as a stochastic process. Consequently, the cost of load shedding is considered. In order to define the optimal power allocation for each generator, the proposed methodology uses Group SO (3) orthogonal matrices (Lies algebra), the marginal costs of the generators, the customer damage cost and Monte-Carlo trials. The result contains generation, marginal cost and load shedding statistics, among others.

Predictive torque control of a multi-drive system fed by a six-leg indirect matrix converter

This paper proposes a predictive torque and flux control for a multi-motor drive system. The scheme is based on a six-leg indirect matrix converter used to drive two induction machines operating at the same speed under different load torque conditions. By using a mathematical model of the converter and machines, the proposed control scheme selects the switching state that minimizes error in the torque and flux predictions according to their reference values. Through simulation results it is shown that the predictive approach can be easily implemented with a good tracking of the controlled variables to their respective references, verifying the fast dynamic response with a good torque tracking conditions and flux regulation in both machines.

Predictive current control of a four-leg indirect matrix converter with imposed source currents and common-mode voltage reduction

This paper presents a new control strategy for a four-leg indirect matrix converter that effectively mitigates common-mode voltages and gives optimal control of source and load currents. This method uses the commutation state of the converter in the subsequent sampling time according to an optimization algorithm given by a simple cost function and a discrete system model. The control goals are the regulation of the output current according to an arbitrary reference and tracking of the source current reference, which is imposed in order to obtain sinusoidal waveforms with low distortion. The technique is enhanced by a reduction of the common-mode voltage using an extra term in the cost function to reduce early motor winding failure and bearing deterioration. Simulation results are presented to support the theoretical development.