Insights into amebiasis using a human 3D‐intestinal model

Entamoeba histolytica is the causative agent of amebiasis, an infectious disease targeting the intestine and the liver in humans. Two types of intestinal infection are caused by this parasite: silent infection, which occurs in the majority of cases, and invasive disease, which affects 10% of infected persons. To understand the intestinal pathogenic process, several in vitro models, such as cell cultures, human tissue explants or human intestine xenografts in mice, have been employed. Nevertheless, our knowledge on the early steps of amebic intestinal infection and the molecules involved during human–parasite interaction is scarce, in part due to limitations in the experimental settings. In the present work, we took advantage of tissue engineering approaches to build a three‐dimensional (3D)‐intestinal model that is able to replicate the general characteristics of the human colon. This system consists of an epithelial layer that develops tight and adherens junctions, a mucus layer and a lamina propria‐like compartment made up of collagen containing macrophages and fibroblast. By means of microscopy imaging, omics assays and the evaluation of immune responses, we show a very dynamic interaction between E. histolytica and the 3D‐intestinal model. Our data highlight the importance of several virulence markers occurring in patients or in experimental models, but they also demonstrate the involvement of under described molecules and regulatory factors in the amoebic invasive process.     

Towards a model for force predictions in the human shoulder.

In this paper the concept of a three-dimensional biomechanical model of the human shoulder is introduced. This model is used to analyze static load sharing between the muscles, the bones and the ligaments. The model consists of all shoulder structures, which means that different positions and different load situations may be analyzed using the same model. Solutions can be found for the complete range of shoulder motion. However, this article focuses only on elevation in the scapular plane and on forces in structures attached to the humerus. The intention is to expand the model in future studies to also involve the forces acting on the other shoulder bones: the scapula and the clavicle. The musculoskeletal forces in the shoulder complex are predicted utilizing the optimization technique with the sum of squared muscle stresses as an objective function. Numerical results predict that among the muscles crossing the glenohumeral joint parts of the deltoideus, the infraspinatus, the supraspinatus, the subscapularis, the pectoralis major, the coracobrachialis and the biceps are the muscles most activated during this sort of abduction. Muscle-force levels reached values of 150 N when the hand load was 1 kg. The results from the model seem to be qualitatively accurate, but it is concluded that in the future development of the model the direction of the contact force in the glenohumeral joint must be constrained.     

The Xenopus retinal ganglion cell as a model neuron to study the establishment of neuronal connectivity

Neurons receive inputs through their multiple branched dendrites and pass this information on to the next neuron via long axons, which branch within the target. The shape the neuron acquires is thus the key to its proper functioning in the neural circuit in which it participates. Both axons and dendrites grow in a directed fashion to their target partner neurons by responding to a large number of molecular cues in the milieu through which they extend. They then go through the process of synaptogenesis, first choosing a neuron on which to synapse, and then the appropriate subcellular location. How a neuron acquires its unique shape, establishes and modifies appropriate synaptic connectivity, and the molecular signals involved, are key questions in developmental neurobiology. Such questions of nervous system wiring are being pursued actively with a variety of different animal models and neuron types, each with its own unique advantages. Among these, the developing retinal ganglion cell (RGC) of the South African clawed frog, Xenopus laevis, has proven particularly fruitful for revealing the secrets of how axons and dendrites acquire their final morphology and connectivity. In this review, we describe how this system can be used to understand the multiple molecular events that instruct the incorporation of RGCs into the neural circuit that controls vision.     

Experimental muscular amebiasis in hamsters as a biological model.

Trophozoites of Entamoeba histolytica maintained in vitro in Pavlova's medium were inoculated by deep intramuscular injection into the proximal left hindleg of hamsters. Thioglycollate medium was utilized as a successful vehicle to induce the infection. The invasion of the muscular tissue by the vegetative forms caused the formation of abscesses with great destruction of muscular fibers. The lesions were limited to the muscular tissue of the femoral area. The number of trophozoites, the medium of thioglycollate as a vehicle, the volume of the inoculum and the trauma caused by the needle were important elements in the evolution of the muscular amebic abscesses. A limited trial of the amebicidal activity of metronidazole utilizing the amebic intramuscular infection was also performed.     

Towards a quantitative model of the post-synaptic proteome

The postsynaptic compartment of the excitatory glutamatergic synapse contains hundreds of distinct polypeptides with a wide range of functions (signalling, trafficking, cell-adhesion, etc.). Structural dynamics in the post-synaptic density (PSD) are believed to underpin cognitive processes. Although functionally and morphologically diverse, PSD proteins are generally enriched with specific domains, which precisely define the mode of clustering essential for signal processing. We applied a stochastic calculus of domain binding provided by a rule-based modelling approach to formalise the highly combinatorial signalling pathway in the PSD and perform the numerical analysis of the relative distribution of protein complexes and their sizes. We specified the combinatorics of protein interactions in the PSD by rules, taking into account protein domain structure, specific domain affinity and relative protein availability. With this model we interrogated the critical conditions for the protein aggregation into large complexes and distribution of both size and composition. The presented approach extends existing qualitative protein-protein interaction maps by considering the quantitative information for stoichiometry and binding properties for the elements of the network. This results in a more realistic view of the postsynaptic proteome at the molecular level.     

Towards a mathematical model of the aortic reservoir

It is well known from experiments that the circulation is going on under physiological parameters, only if the rigidity of the aortic wall is sufficiently small. We show that the result can be confirmed by the one-dimensional mathematical model describing the dynamics of the aortic reservoir (AR) by means of a hyperbolic system of PDE.     

Towards a predictive systems-level model of the human microbiome: progress,challenges, and opportunities

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Percutaneous cryoablation of the kidney in a porcine model

Several reports have documented that renal cryoablation when used either via open or laparoscopic surgical techniques can be well tolerated in animal models. We sought to examine the feasibility of performing renal cryoablation percutaneously, under ultrasound guidance, in a porcine model. A total of 13 juvenile swine was treated. In each animal the lower pole of the left kidney was used as the target. Each animal had serum drawn for Hct, Wbc, CK, BUN, creatinine, and myoglobin pre- and postcryoablation and again at the time of sacrifice. Animals were sacrificed at 0-42 days postoperatively, and the treated renal units were harvested and submitted for histologic examination. The procedures were divided into three groups based on the extent of the freeze time. Group 1 (n = 4) was treated for 3-4 min, single freeze; Group 2 (n = 3), 6-7 min, single freeze; and Group 3 (n = 6), a double freeze-thaw cycle, each for 10 min. None of the animals in this experiment died or experienced significant clinical deteriorations at any time post-treatment. No significant differences in any of the measured serum markers were noted between pre- and post-treatment values. For animals in Groups 1 and 2 discrete cryolesions were created with sharp 1-mm borders and no perirenal reaction/damage to surrounding structures. For animals in Group 3 large areas of renal infarction/cryolesions were produced, with significant perirenal reaction in 5/6 animals and gross hydronephrosis/total renal loss in 2/6. Percutaneous renal cryoablation appears to be well tolerated in the porcine model which we used. Associated morbidity appears to be related to the extent of freezing, with a safety tolerance in the present study limited to target areas of approximately 3-5 cm3. These findings suggest that a pilot study of percutaneous renal cryoablation for patients with 3-4-cm exophytic lesions may be warranted.     

Towards personalized diagnostics via longitudinal study of the human plasma N-glycome

Facilitated by substantial advances in analytical methods, plasma N-glycans have emerged as potential candidates for biomarkers. In the recent years, several investigations could link aberrant plasma N-glycosylation to numerous diseases. However, due to often limited specificity and sensitivity, only a very limited number of glycan biomarkers were approved by the authorities up to now. The inter-individual heterogeneity of the plasma N-glycomes might mask disease related changes in conventional large cross-sectional cohort studies, with a one-time sampling approach. But, a possible benefit of longitudinal sampling in biomarker discovery could be, that already small changes during disease progression are revealed, by monitoring the plasma N-glycome of individuals over time.To evaluate this, we collected blood plasma samples of five healthy donors over a time period of up to six years (min. 1.5 years). The plasma N-glycome was analyzed by xCGE-LIF, to investigate the intra-individual N-glycome variability over time. It is shown, that the plasma N-glycome of an individual is remarkably stable over a period of several years, and that observed small longitudinal changes are independent from seasons, but significantly correlated with lifestyle and environmental factors. Thus, the potential of future longitudinal biomarker discovery studies could be demonstrated, which is a further step towards personalized diagnostics. This article is part of a Special Issue entitled “Glycans in personalised medicine” Guest Editor: Professor Gordan Lauc.     

Ingraft chimerism in lung transplantation - a study in a porcine model of obliterative bronchiolitis

Background

Bronchial epithelium is a target of the alloimmune response in lung transplantation, and intact epithelium may protect allografts from rejection and obliterative bronchiolitis (OB). Herein we study the influence of chimerism on bronchial epithelium and OB development in pigs.

Methods

A total of 54 immunosuppressed and unimmunosuppressed bronchial allografts were serially obtained 2-90 days after transplantation. Histology (H&E) was assessed and the fluorescence in situ hybridization (FISH) method for Y chromosomes using pig-specific DNA-label was used to detect recipient derived cells in graft epithelium and bronchial wall, and donor cell migration to recipient organs. Ingraft chimerism was studied by using male recipients with female donors, whereas donor cell migration to recipient organs was studied using female recipients with male donors.

Results

Early appearance of recipient-derived cells in the airway epithelium appeared predictive of epithelial destruction (R = 0.610 - 0.671 and p < 0.05) and of obliteration of the bronchial lumen (R = 0.698 and p < 0.01). All allografts with preserved epithelium showed epithelial chimerism throughout the follow-up. Antirejection medication did not prevent, but delayed the appearance of Y chromosome positive cells in the epithelium (p < 0.05), or bronchial wall (p < 0.05).

Conclusions

In this study we demonstrate that early appearance of Y chromosomes in the airway epithelium predicts features characteristic of OB. Chimerism occurred in all allografts, including those without features of OB. Therefore we suggest that ingraft chimerism may be a mechanism involved in the repair of alloimmune-mediated tissue injury after transplantation.     

Towards a rational modeling for the human placenta

An attempt is made to provide a general framework sufficient of handling analytically typical questions concerned with a physiological functioning of a human placenta. The principles of mechanics and thermodynamics, as they apply to a continuum theory of mixtures, are the essential tools in constructing the model. In general terms, it is assumed that each point of the placenta space is simultaneously occupied by placenta tissue, fetal blood, and maternal blood. These three constituents are allowed to interact mechanically, chemically, and thermally. One of the basic features of the model is its capability of providing quantitative information on the deformation of the placenta tissue and the pressure distribution of the fetal and maternal blood. Effects of inertia, deformation, and changes of shape and weight of the placenta during gestation are also taken into account.     

Towards a unified model of neuroendocrine-immune interaction

Although the neuroendocrine system has immunomodulating potential, studies examining the relationship between stress, immunity and infection have, until recently, largely been the preserve of behavioural psychologists. Over the last decade, however, immunologists have begun to increasingly appreciate that neuroendocrine-immune interactions hold the key to understanding the complex behaviour of the immune system in vivo. The nervous, endocrine and immune systems communicate bidirectionally via shared messenger molecules variously called neurotransmitters, cytokines or hormones. Their classification as neurotransmitters, cytokines or hormones is more serendipity than a true reflection of their sphere of influence. Rather than these systems being discrete entities we would propose that they constitute, in reality, a single higher-order entity. This paper reviews current knowledge of neuroendocrine-immune interaction and uses the example of T-cell subset differentiation to show the previously under-appreciated importance of neuroendocrine influences in the regulation of immune function and, in particular, Th1/Th2 balance and diurnal variation there of.     

Induction of the IFN-gamma gene and protein is linked to the establishment of cell polarity in a porcine epithelial cell line

We report here original properties of a porcine trophectoderm cell line, TBA B4-3, that developed a polarized phenotype with high transepithelial electrical resistance (TER) values and functional tight junctions (TJs) when grown on a microporous membrane. We found that treatment of polarized TBA B4-3 cells with a strong protein kinase C (PKC) agonist, phorbol 12-myristate-13-acetate (PMA), induced 3-4 days later a transient interferon-gamma (IFN-gamma) mRNA expression and vectorial IFN-gamma protein secretion toward the apical side of the monolayer. Exposure of TBA B4-3 cells to PMA first resulted in a rapid and profound disorganization of the monolayer structure mainly characterized by the appearance of multilayered polyp-like foci structures, a strong decrease of the TER, and a increase of permeability correlated with changes in the organization and localization of the TJ-associated proteins (ZO-1 and occludin) and filamentous actin (f-actin). After PMA removal, spontaneous return to the initial polarized monolayer state occurred, characterized by TER rising to prestimulation values, TJ protein relocalization, and multilayered cell structures fading. This return was strictly correlated with transient IFN-gamma gene induction. Our report represents the first example of an inducible expression of IFN-gamma by a polarized epithelial cell. After PMA treatment, the close correlation between establishment of cell polarity and IFN-gamma gene expression suggests a link between these phenomena. This also suggests a novel biological mechanism by which transient and reversible disorganization of a polarized monolayer of epithelial cells could trigger regulated expression of a cytokine gene by these cells.     

Towards a psycho-physiological model of thermal perception

Recommendations for indoor thermal requirements have been based upon verbalized responses on traditional assumptions that (1) minimal thermoregulatory activity may be equated to maximum subjective acceptability (2) sensations and levels of discomfort are synonymous and (3) perception of warmth is exclusively the function of thermal stimulus — physiological response. These concepts are reviewed in the light of recent researches which indicate the inadequacy of the existing physiological models and methods of research. In particular, recognition is made of higher levels of mental integration of information flows which, it is argued, must include parameters of past cultural and climatic experiences and expectations. The aim is to initiate a more holistic approach to research into human thermal environments, and, a clearer definition of concepts significant to practical application.