Trogocytosis by Entamoeba histolytica Mediates Acquisition and Display of Human Cell Membrane Proteins and Evasion of Lysis by Human Serum.

We previously showed that Entamoeba histolytica kills human cells through a mechanism that we termed trogocytosis ("trogo-" means "nibble"), due to its resemblance to trogocytosis in other organisms. In microbial eukaryotes like E. histolytica, trogocytosis is used to kill host cells. In multicellular eukaryotes, trogocytosis is used for cell killing and cell-cell communication in a variety of contexts. Thus, nibbling is an emerging theme in cell-cell interactions both within and between species. When trogocytosis occurs between mammalian immune cells, cell membrane proteins from the nibbled cell are acquired and displayed by the recipient cell. In this study, we tested the hypothesis that through trogocytosis, amoebae acquire and display human cell membrane proteins. We demonstrate that E. histolytica acquires and displays human cell membrane proteins through trogocytosis and that this leads to protection from lysis by human serum. Protection from human serum occurs only after amoebae have undergone trogocytosis of live cells but not phagocytosis of dead cells. Likewise, mutant amoebae defective in phagocytosis, but unaltered in their capacity to perform trogocytosis, are protected from human serum. Our studies are the first to reveal that amoebae can display human cell membrane proteins and suggest that the acquisition and display of membrane proteins is a general feature of trogocytosis. These studies have major implications for interactions between E. histolytica and the immune system and also reveal a novel strategy for immune evasion by a pathogen. Since other microbial eukaryotes use trogocytosis for cell killing, our findings may apply to the pathogenesis of other infections.IMPORTANCE Entamoeba histolytica causes amoebiasis, a potentially fatal diarrheal disease. Abscesses in organs such as the liver can occur when amoebae are able to breach the intestinal wall and travel through the bloodstream to other areas of the body. Therefore, understanding how E. histolytica evades immune detection is of great interest. Here, we demonstrate for the first time that E. histolytica acquires and displays human cell membrane proteins by taking "bites" of human cell material in a process named trogocytosis ("trogo-" means "nibble"), and that this allows amoebae to survive in human serum. Display of acquired proteins through trogocytosis has been previously characterized only in mammalian immune cells. Our study suggests that this is a more general feature of trogocytosis not restricted to immune cells and broadens our knowledge of eukaryotic biology. These findings also reveal a novel strategy for immune evasion by a pathogen and may apply to the pathogenesis of other infections

Trypanin, a Component of the Flagellar Dynein Regulatory Complex, Is Essential in Bloodstream Form African Trypanosomes

The Trypanosoma brucei flagellum is a multifunctional organelle with critical roles in motility, cellular morphogenesis, and cell division. Although motility is thought to be important throughout the trypanosome lifecycle, most studies of flagellum structure and function have been restricted to the procyclic lifecycle stage, and our knowledge of the bloodstream form flagellum is limited. We have previously shown that trypanin functions as part of a flagellar dynein regulatory system that transmits regulatory signals from the central pair apparatus and radial spokes to axonemal dyneins. Here we investigate the requirement for this dynein regulatory system in bloodstream form trypanosomes. We demonstrate that trypanin is localized to the flagellum of bloodstream form trypanosomes, in a pattern identical to that seen in procyclic cells. Surprisingly, trypanin RNA interference is lethal in the bloodstream form. These knockdown mutants fail to initiate cytokinesis, but undergo multiple rounds of organelle replication, accumulating multiple flagella, nuclei, kinetoplasts, mitochondria, and flagellum attachment zone structures. These findings suggest that normal flagellar beat is essential in bloodstream form trypanosomes and underscore the emerging concept that there is a dichotomy between trypanosome lifecycle stages with respect to factors that contribute to cell division and cell morphogenesis. This is the first time that a defined dynein regulatory complex has been shown to be essential in any organism and implicates the dynein regulatory complex and other enzymatic regulators of flagellar motility as candidate drug targets for the treatment of African sleeping sickness

Hadron Helicity Violation in Exclusive Processes: Quantitative Calculations in Leading Order QCD

We study a new mechanism for hadronic helicity flip in high energy hard exclusive reactions. The mechanism proceeds in the limit of perfect chiral symmetry, namely without any need to flip a quark helicity. The fundamental feature of the new mechanism is the breaking of rotational symmetry of the hard collision by a scattering plane in processes involving independent quark scattering. We show that in the impulse approximation there is no evidence for of the helicity violating process as the energy or momentum transfer $Q^2$ is increased over the region 1 GeV^2 < Q^2 < 100 GeV^2. In the asymptotic region Q^2> 1000 GeV^2, a saddle point approximation with doubly logarithmic accuracy yields suppression by a fraction of power of Q^2. ``Chirally--odd" exclusive wave functions which carry non--zero orbital angular momentum and yet are leading order in the high energy limit, play an important role.Comment: uuencoded LaTeX file (21 pages) and PostScript figure

Corrosion Inhibition of Aluminum Alloy 2024-T3 by Aqueous Vanadium Species

Nuclear magnetic resonance (NMR) measurements were made on aqueous vanadate solutions to characterize speciation as a function of pH and vanadate concentration. Additionally, potentiodynamic polarization measurements were carried out on Al alloy 2024-T3 in 50 mM NaCl solutions in which pH and vanadate concentration were systematically varied. Results showed that inhibition by vanadates occurred mainly in alkaline solutions where tetrahedrally coordinated vanadates, metavanadate and pyrovanadate, were abundant. Inhibition was not observed in solutions where octahedrally coordinated decavanadates predominated. Anodic inhibition, in the form of increased pitting potential, was observed in both aerated and deaerated solutions. In contrast, cathodic inhibition was observed only in aerated solutions acting primarily through the suppression of oxygen reduction. Energy-dispersive spectroscopy, used to collect chemical maps from aluminum coupons exposed to vanadate solutions, showed the suppression of Al_2CuMg particle dissolution compared to vanadate-free solutions. NMR measurements were also used to track changes in vanadate speciation with time, pH adjustment, and with exposure to metallic aluminum surfaces. NMR showed noninhibiting octahedrally coordinated decavanadates rapidly decompose into inhibiting tetrahedrally coordinated metavanadates and pyrovanadates after alkaline pH adjustment. While decomposition begins immediately upon pH adjustment, equilibrium may not be reached even after significant time periods

Exclusive Hadronic Processes and Color Transparency

We review the current status of high energy exclusive processes and color transparency.Comment: 17 pages, 8 figures, based on talk given at International Symposium on Nuclear Physics, Mumbai, Dec 18-22, 200

MEF2C regulates outflow tract alignment and transcriptional control of Tdgf1

Congenital heart defects are the most common birth defects in humans, and those that affect the proper alignment of the outflow tracts and septation of the ventricles are a highly significant cause of morbidity and mortality in infants. A late differentiating population of cardiac progenitors, referred to as the anterior second heart field (AHF), gives rise to the outflow tract and the majority of the right ventricle and provides an embryological context for understanding cardiac outflow tract alignment and membranous ventricular septal defects. However, the transcriptional pathways controlling AHF development and their roles in congenital heart defects remain incompletely elucidated. Here, we inactivated the gene encoding the transcription factor MEF2C in the AHF in mice. Loss of Mef2c function in the AHF results in a spectrum of outflow tract alignment defects ranging from overriding aorta to double-outlet right ventricle and dextro-transposition of the great arteries. We identify Tdgf1, which encodes a Nodal co-receptor (also known as Cripto), as a direct transcriptional target of MEF2C in the outflow tract via an AHFrestricted Tdgf1 enhancer. Importantly, both the MEF2C and TDGF1 genes are associated with congenital heart defects in humans. Thus, these studies establish a direct transcriptional pathway between the core cardiac transcription factor MEF2C and the human congenital heart disease gene TDGF1. Moreover, we found a range of outflow tract alignment defects resulting from a single genetic lesion, supporting the idea that AHF-derived outflow tract alignment defects may constitute an embryological spectrum rather than distinct anomalies

Multiple-Scattering Series For Color Transparency

Color transparency CT depends on the formation of a wavepacket of small spatial extent. It is useful to interpret experimental searches for CT with a multiple scattering scattering series based on wavepacket-nucleon scattering instead of the standard one using nucleon-nucleon scattering. We develop several new techniques which are valid for differing ranges of energy. These techniques are applied to verify some early approximations; study new forms of the wave-packet-nucleon interaction; examine effects of treating wave packets of non-zero size; and predict the production of $N^*$'s in electron scattering experiments.Comment: 26 pages, U.Wa. preprint 40427-23-N9

Time reversal in thermoacoustic tomography - an error estimate

The time reversal method in thermoacoustic tomography is used for approximating the initial pressure inside a biological object using measurements of the pressure wave made on a surface surrounding the object. This article presents error estimates for the time reversal method in the cases of variable, non-trapping sound speeds.Comment: 16 pages, 6 figures, expanded "Remarks and Conclusions" section, added one figure, added reference

Local Food Systems: Concepts, Impacts, and Issues

This comprehensive overview of local food systems explores alternative definitions of local food, estimates market size and reach, describes the characteristics of local consumers and producers, and examines early indications of the economic and health impacts of local food systems. There is no consensus on a definition of “local” or “local food systems” in terms of the geographic distance between production and consumption. But defining “local” based on marketing arrangements, such as farmers selling directly to consumers at regional farmers’ markets or to schools, is well recognized. Statistics suggest that local food markets account for a small, but growing, share of U.S. agricultural production. For smaller farms, direct marketing to consumers accounts for a higher percentage of their sales than for larger farms. Findings are mixed on the impact of local food systems on local economic development and better nutrition levels among consumers, and sparse literature is so far inconclusive about whether localization reduces energy use or greenhouse gas emissions.local food systems, farmers’ markets, direct-to-consumer marketing, direct-to-retail/ foodservice marketing, community supported agriculture, farm to school programs, Farmers’ Market Promotion Program, food miles, Community/Rural/Urban Development,