Design and interpretation of cell trajectory assays

Cell trajectory data are often reported in the experimental cell biology literature to distinguish between different types of cell migration. Unfortunately, there is no accepted protocol for designing or interpreting such experiments and this makes it difficult to quantitatively compare different published datasets and to understand how changes in experimental design influence our ability to interpret different experiments. Here, we use an individual-based mathematical model to simulate the key features of a cell trajectory experiment. This shows that our ability to correctly interpret trajectory data is extremely sensitive to the geometry and timing of the experiment, the degree of motility bias and the number of experimental replicates. We show that cell trajectory experiments produce data that are most reliable when the experiment is performed in a quasi-one-dimensional geometry with a large number of identically prepared experiments conducted over a relatively short time-interval rather than a few trajectories recorded over particularly long time-intervals.     

Ultrasmall Nanoplatforms as Calcium‐Responsive Contrast Agents for Magnetic Resonance Imaging

The preparation of ultrasmall and rigid platforms (USRPs) that are covalently coupled to macrocycle‐based, calcium‐responsive/smart contrast agents (SCAs), and the initial in vitro and in vivo validation of the resulting nanosized probes (SCA‐USRPs) by means of magnetic resonance imaging (MRI) is reported. The synthetic procedure is robust, allowing preparation of the SCA‐USRPs on a multigram scale. The resulting platforms display the desired MRI activity—i.e., longitudinal relaxivity increases almost twice at 7 T magnetic field strength upon saturation with Ca²⁺. Cell viability is probed with the MTT assay using HEK‐293 cells, which show good tolerance for lower contrast agent concentrations over longer periods of time. On intravenous administration of SCA‐USRPs in living mice, MRI studies indicate their rapid accumulation in the renal pelvis and parenchyma. Importantly, the MRI signal increases in both kidney compartments when CaCl₂ is also administrated. Laser‐induced breakdown spectroscopy experiments confirm accumulation of SCA‐USRPs in the renal cortex. To the best of our knowledge, these are the first studies which demonstrate calcium‐sensitive MRI signal changes in vivo. Continuing contrast agent and MRI protocol optimizations should lead to wider application of these responsive probes and development of superior functional methods for monitoring calcium‐dependent physiological and pathological processes in a dynamic manner.     

LAIR1, an ITIM-Containing Receptor Involved in Immune Disorders and in Hematological Neoplasms

Leukocyte-associated immunoglobulin (Ig)-like receptor 1 (LAIR1, CD305) belongs to the family of immune-inhibitory receptors and is widely expressed on hematopoietic mature cells, particularly on immune cells. Four different types of ligands of LAIR1 have been described, including collagens, suggesting a potential immune-regulatory function on the extracellular matrix. By modulating cytokine secretion and cellular functions, LAIR1 displays distinct patterns of expression among NK cell and T/B lymphocyte subsets during their differentiation and cellular activation and plays a major negative immunoregulatory role. Beyond its implications in physiology, the activity of LAIR1 can be inappropriately involved in various autoimmune or inflammatory disorders and has been implicated in cancer physiopathology, including hematological neoplasms. Its action as an inhibitory receptor can result in the dysregulation of immune cellular responses and in immune escape within the tumor microenvironment. Furthermore, when expressed by tumor cells, LAIR1 can modulate their proliferation or invasion properties, with contradictory pro- or anti-tumoral effects depending on tumor type. In this review, we will focus on its role in normal physiological conditions, as well as during pathological situations, including hematological malignancies. We will also discuss potential therapeutic strategies targeting LAIR1 for the treatment of various autoimmune diseases and cancer settings.     

Tie-Over Bolster Pressure Dressing Improves Outcomes of Skin Substitutes Xenografts on Athymic Mice

The efficacy of skin substitutes is established for the treatment of burn injuries, but its use is not limited to this condition. This technology has the potential to improve the treatment of various conditions by offering highly advanced and personalized treatments. In vivo studies are challenging but essential to move to clinical use in humans. Mice are the most widely used species in preclinical studies, but the main drawback of this model is the limited surface area of the graft in long-term transplantation studies caused by the displacement and the contraction of the graft. We improved the conventional surgical procedures by stabilizing the chamber covering the graft with intramuscular sutures and by adding a tie-over bolster dressing. The current study was therefore performed to compare outcomes of skin grafts between the conventional and optimized skin graft model. Human self-assembled skin substitutes (SASSs) were prepared and grafted to athymic mice either by the conventional method or by the new grafting method. Graft healing and complications were assessed using digital photographs on postoperative days 7, 14, and 21. Similar structure and organization were observed by histological staining. The new grafting method reduced medium and large displacement events by 1.26-fold and medium and large contraction events by 1.8-fold, leading to a 1.6-fold increase in graft surface area compared to skin substitutes grafted with the usual method. This innovation ensures better reproducibility and consistency of skin substitute transplants on mice.     

An E3‐14.7K Peptide that Promotes Microtubules‐Mediated Transport of Plasmid DNA Increases Polyplexes Transfection Efficiency

Chemical vectors as cationic polymers and cationic lipids are promising alternatives to viral vectors for gene therapy. Beside endosome escape and nuclear import, plasmid DNA (pDNA) migration in the cytosol toward the nuclear envelope is also regarded as a limiting step for efficient DNA transfection with non‐viral vectors. Here, the interaction between E3‐14.7K and FIP‐1 to favor migration of pDNA along microtubules is exploited. E3‐14.7K is an early protein of human adenoviruses that interacts via FIP‐1 (Fourteen.7K Interacting Protein 1) protein with the light‐chain components of the human microtubule motor protein dynein (TCTEL1). This peptide is conjugated with pDNA and mediates interaction of pDNA in vitro with isolated microtubules as well as with microtubules in cellulo. Videomicroscopy and tracking treatment of images clearly demonstrate that P79‐98/pDNA conjugate exhibits a linear transport with large amplitude along microtubules upon 2 h transfection with polyplexes whereas control pDNA conjugate exhibits small non‐directional movements in the cytoplasm. Remarkably, P79‐98/peGFP polyplexes enhance by a factor 2.5 (up to 76%) the number of transfected cells. The results demonstrate, for the first time, that the transfection efficiency of polyplexes can be drastically increased when the microtubules migration of pDNA is facilitated by a peptide allowing pDNA docking to TCTEL1. This is a real breakthrough in the non viral gene delivery field that opens hope to build artificial viruses.     

Process optimization in the self-reacting friction stir welding of aluminum 6061-T6

Self-reacting friction stir welding (SR-FSW), also called bobbin-tool friction stir welding (BT-FSW), is a solid state welding process similar to friction stir welding (FSW) except that the tool has two opposing shoulders instead of the shoulder and a backing plate found in FSW. The tool configuration results in greater heat input and a symmetrical weld macrostructure. A significant amount of information has been published in the literature concerning traditional FSW while little has been published about SR-FSW. An optimization experiment was performed using a factorial design to evaluate the effect of process parameters on the weld temperature, surface and internal quality, and mechanical properties of self-reacting friction stir welded aluminum alloy 6061-T6 butt joints. The parameters evaluated were tool rotational speed, traverse speed, and tool plunge force. A correlation between weld temperature, defect formation (specifically galling and void formation), and mechanical properties was found. Optimum parameters were determined for the welding of 8-mm-thick 6061-T6 plate.     

Essential fatty acids in trout serum lipoproteins,vitellogenin and egg lipids

This paper describes evidence of (n−3) and particularly of 22∶6 (n−3) fatty acid enrichment in trout lipoproteins as well as in vitellogenin, egg lipovitellin and oil globule. Among the lipoproteins, HDL and LDL were the main forms of blood lipid transport, whereas phospholipids and cholesteryl esters are the preferential chemical carriers for (n−3) fatty acid transport. However, cholesteryl esters were less important as esterified fatty acid carriers than in man. Taken together with the data obtained in mammals, our results suggest that there may be a relationship between EFA activity and the distribution of the EFA among the lipoprotein lipid fractions in vertebrates, irrespective of the EFA series. Administration of an (n−3) fatty acid deficient diet for three months prior to trout spawning produced a significant increase in egg lipid content, primarily as a result of the increase of the oil globule composed almost exclusively of triacylglycerols. This diet decreased the 22∶6 (n−3), as well as the (n−3) fatty acid contents of lipoproteins, lipovitellin, vitellogenin and the oil globule. In contrast, the (n−3) fatty acid level was always higher in lipoproteins and lipovitellin than in the vitellogenin and the oil globule. Moreover, the relative levels of 22∶6 (n−3) and total (n−3) fatty acids were quite similar in lipoproteins and lipovitellin on the one hand, and in vitellogenin and the oil globule on the other. These findings suggest a direct relationship between the two forms of plasma lipid transport and the two egg compartments. During ovogenesis, dietary lipids seemed to be diverted from the adipose tissue and essentially deposited in the egg.     

évaluation du développement de l'enfant au cours de la première année: l'utilisation de regroupements d'items du Bayley.

Compared the discriminative powers of (1) the mental and motor indexes of the Bayley Scales of Infant Development (BSID) (N. Bayley, 1969), (2) the 5-cluster regrouping of the BSID by R. Kohen-Raz (1967), and (3) the 7-cluster regrouping of the BSID by L. J. Yarrow et al (1975). Ss were 29 3.5-mo-old, 53 6-mo-old, and 57 9-mo-old Haitian, Canadian, and Vietnamese infants. Ss' performances on the BSID were analyzed with the mental and motor scales of the BSID, the 5-cluster regrouping, and the 7-cluster regrouping. The 3 systems were compared with regard to their ability to detect age, gender, and cultural differences. (English abstract) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The social patterning of injury repetitions among young car drivers in Sweden

A national register-based cohort study was conducted to explore whether the social patterning of car drivers suffering injury repetitions differs from that of once-injured drivers in a cohort of young persons in Sweden (aged 18-26). Injury repeaters were defined as individuals sustaining injuries as a car driver on more than one occasion over an 8-year period. Only subjects obtaining a driver's licence before the age of 27 were included in the study. The social variables considered were, in turn, gender, socioeconomic position of origin, and own educational attainment. Attention was also paid to age at licensing. Two types of comparisons were made, using odds ratios with 95% confidence intervals. First, the odds of injury repeaters were computed using the once-injured car drivers as a reference group. Second, odds were compiled for once, twice, and three or more times injured people, using the non-injured at all as a comparison group. The results show that, by and large, the injury-risk distribution of car drivers with injury repetition does not differ from that of one-injury drivers with regard to gender, education, or socioeconomic group. However, drivers from self-employed households show greater odds of injury repetition (OR 1.65, CI 1.02-2.67) than of one injury compared with drivers from the families of non-manual employees. Since the number of injury repeaters is low and their socioeconomic distribution is very similar to that of the once injured, there is no need to regard them as a group at excess risk or different from the one-time injured. Reducing risk levels and risk differentials for the one time injured should therefore receive priority with regard to traffic-injury prevention.