Limitations in the methods employed to generate micrometric colloidal droplets hinder the emergence of key applications in the fields of material science and drug delivery. Through the use of dedicated nanofluidic devices and by taking advantage of an original physical effect called capillary focusing, we could circumvent some of these limitations. The nanofluidic (i.e., submicrometric) devices introduced herein are made of soft materials, and their fabrication relies upon rapid technologies. The objects that we have generated are simple droplets, multiple droplets, particles, and Janus particles whose sizes lie between 900 nm and 3 mum (i.e., within the colloidal range). Colloidal droplets have been assembled on-chip into clusters and crystals, yielding discrete diffraction patterns. We illustrate potential applications in the field of drug delivery by demonstrating the ability of multiple droplets to be phagocytosed by murine macrophage-type cells.

Background: In Europe, sensitization to ash pollen induces pollinosis with cross-reactivities with other pollen sources. The aim of the study was to identify the repertoire of ash pollen allergens and evaluate the extent of the diversity of the IgE response in ash allergic patients. Methods: The IgE reactivities of 114 ash pollen- and eight grass pollen-sensitized patients were screened by 1D immunoblot (SDS-PAGE) against ash pollen extract. The IgE reactivities of 13 ash pollen- and two grass pollen-sensitized patients were then evaluated in 2D immunoblots. Some IgE- and non-IgE-reactive proteins were identified by mass spectrometry. Results: In 1D analysis, 86% of sera showed binding to Fra e 1 (18-20 kDa), 23% to Fra e 2 (14 kDa), 3% to Fra e 3 (10 kDa) and 57% to High Molecular Weight allergens (HMW,>30 kDa). Individual analysis of 2D immunoblots showed several IgE-binding protein areas among which three were more often recognized:(i) Fra e 1 comprising, at least, 15 isoforms,(ii) a series of acidic spots (45 kDa), and (iii) Fra e 2, the ash profilin. HMW allergens could be resolved in four areas; two unidentified, one homologous to beta-galactosidase and the other to sugar transport proteins. A malate deshydrogenase and calmodulin were shown to be IgE-binding proteins and 10 non-IgE reactive proteins were identified. Conclusions: No direct correlation was evidenced between IgE profile and the degree of sensitization even though 2 spectrotypes could be distinguished. Our data contribute to a better delineation of ash pollen allergens and patterns of sensitization.

Summary White coat colour in horses is inherited as a monogenic autosomal dominant trait showing a variable expression of coat depigmentation. Mutations in the KIT gene have previously been shown to cause white coat colour phenotypes in pigs, mice and humans. We recently also demonstrated that four independent mutations in the equine KIT gene are responsible for the dominant white coat colour phenotype in various horse breeds. We have now analysed additional horse families segregating for white coat colour phenotypes and report seven new KIT mutations in independent Thoroughbred, Icelandic Horse, German Holstein, Quarter Horse and South German Draft Horse families. In four of the seven families, only one single white horse, presumably representing the founder for each of the four respective mutations, was available for genotyping. The newly reported mutations comprise two frameshift mutations (c.1126_1129delGAAC; c.2193delG), two missense mutations (c.856G>A; c.1789G>A) and three splice site mutations (c.338-1G>C; c.2222-1G>A; c.2684+1G>A). White phenotypes in horses show a remarkable allelic heterogeneity. In fact, a higher number of alleles are molecularly characterized at the equine KIT gene than for any other known gene in livestock species.

The KIT receptor protein-tyrosine kinase plays an important role during embryonic development. Activation of KIT is crucial for the development of various cell lineages such as melanoblasts, stem cells of the haematopoietic system, spermatogonia and intestinal cells of Cajal. In mice, many mutations in the Kit gene cause pigmentation disorders accompanied by pleiotropic effects on blood cells and male fertility. Previous work has demonstrated that dominant white Franches-Montagnes horses carry one copy of the KIT gene with the p.Y717X mutation. The targeted breeding of white horses would be ethically questionable if white horses were known to suffer from anaemia or leukopenia. The present study demonstrates that no statistically significant differences in peripheral blood parameters are detectable between dominant white and solid-coloured Franches-Montagnes horses. The data indicate that KIT mutations may have different effects in mice, pigs, and horses. The KIT p.Y717X mutation does not have a major negative effect on the haematopoietic system of dominant white horses.

White markings and spotting patterns in animal species are thought to be a result of the domestication process. They often serve for the identification of individuals but sometimes are accompanied by complex pathological syndromes. In the Swiss Franches-Montagnes horse population, white markings increased vastly in size and occurrence during the past 30 years, although the breeding goal demands a horse with as little depigmented areas as possible. In order to improve selection and avoid more excessive depigmentation on the population level, we estimated population parameters and breeding values for white head and anterior and posterior leg markings. Heritabilities and genetic correlations for the traits were high (h(2)> 0.5). A strong positive correlation was found between the chestnut allele at the melanocortin-1-receptor gene locus and the extent of white markings. Segregation analysis revealed that our data fit best to a model including a polygenic effect and a biallelic locus with a dominant-recessive mode of inheritance. The recessive allele was found to be the white trait-increasing allele. Multilocus linkage disequilibrium analysis allowed the mapping of the putative major locus to a chromosomal region on ECA3q harboring the KIT gene.

White coat color has been a highly valued trait in horses for at least 2,000 years. Dominant white (W) is one of several known depigmentation phenotypes in horses. It shows considerable phenotypic variation, ranging from approximately 50% depigmented areas up to a completely white coat. In the horse, the four depigmentation phenotypes roan, sabino, tobiano, and dominant white were independently mapped to a chromosomal region on ECA 3 harboring the KIT gene. KIT plays an important role in melanoblast survival during embryonic development. We determined the sequence and genomic organization of the approximately 82 kb equine KIT gene. A mutation analysis of all 21 KIT exons in white Franches-Montagnes Horses revealed a nonsense mutation in exon 15 (c.2151C>G, p.Y717X). We analyzed the KIT exons in horses characterized as dominant white from other populations and found three additional candidate causative mutations. Three almost completely white Arabians carried a different nonsense mutation in exon 4 (c.706A>T, p.K236X). Six Camarillo White Horses had a missense mutation in exon 12 (c.1805C>T, p.A602V), and five white Thoroughbreds had yet another missense mutation in exon 13 (c.1960G>A, p.G654R). Our results indicate that the dominant white color in Franches-Montagnes Horses is caused by a nonsense mutation in the KIT gene and that multiple independent mutations within this gene appear to be responsible for dominant white in several other modern horse populations.

BACKGROUND: Standard modalities to assist in determining the extent of chest wall developmental deformities in patients include x-ray and computed tomography (CT). The purpose of this study is to describe an optical imaging technique that provides accurate cross-sectional images of the chest, and to compare these with standard CT-derived images of chest wall abnormalities. PATIENTS AND METHODS: Ten patients (5 pectus excavatum and 5 pectus carinatum) underwent imaging that included limited CT and optical cross-sectional imaging. Severity indices of the deformity using the standard Haller index (HI) were calculated from CT scans. A similar severity measurement of deformity was derived from the outline of torso cross sections (ie, from skin to skin measurements) obtained from optical images. To assess the severity of carinatum defects, a modified pectus index was derived, which measures the anterior chest protrusion from the central chord of the chest cross section. We performed regression analyses, comparing the indices obtained from CT and optical imaging methodologies. RESULTS: Optical measures of cross-sectional deformities correlated well with standard HI (r(2)= 0.94) and even better with the modified pectus index (r(2)= 0.96). Adaptation of the HI for pectus carinatum deformity evaluation was effective, and consistent with the torso surface deformity measures. CONCLUSIONS: Torso models from optical imaging offer 3-D images of the chest wall deformity with no radiation exposure. This preliminary study showed promising results for the use of torso surface measurement as an alternative index of pectus deformities; if validated in larger studies, these measures may be useful for following chest wall abnormalities, using repeated studies in patients.

A study carried out on 49 horses showed that it is possible to measure the attention time by operant conditioning. After teaching horses an instrumental task using a signal, we were then able to test their attention time by asking them to prolong it increasingly while setting success and failure criteria. Two tests were performed 3 weeks apart. The 2nd test was feasible without relearning, a proof of memory, and was repeatable, a proof of consistency in the attention time. A significant difference was observed between the 3 age groups. Young horses often performed very well during the 1st test but their attention dropped in the 2nd test while older horses were more stable with respect to attention and even increased it slightly. The study shows that there are individual differences but it was not possible to prove a significant influence of breed, gender and paternal influence. Consequently, learning appears to be one of the most interesting approaches for evaluating the attention of horses and for observing their behaviour.

Scoliosis is a common and poorly understood spinal disorder that is clinically monitored with a series of full spinal X-rays. The purpose of this study was to predict scoliosis future progression at 6- and 12-month intervals with successive spinal indices and a hybrid learning technique (i.e., the combination of fuzzy c-means clustering and artificial neural network (ANN)). Ultimately this could decrease scoliotic patients' radiation exposure and the associated cancer risk in growing adolescents. Seventy-two data sets were derived from a database of 56 acquisitions from 11 subjects (29.8 ± 9.6° Cobb angle, 11.4 ± 2.4 yr), each consisting of 4 sequential values of Cobb angle and lateral deviations at apices in 6- and 12-month intervals in the coronal plane. Progression patterns in Cobb angles (n = 10) and lateral deviations (n = 8) were successfully identified using a fuzzy c-means clustering algorithm. The accuracies of the trained ANN, having a structure of three input variables, four nonlinear hidden nodes, and one linear output variable, for training and test data sets were within 3.64°(± 2.58°) and 4.40°(± 1.86°) of Cobb angles, and within 3.59 (± 3.96) mm and 3.98 (± 3.41) mm of lateral deviations, respectively. Those results were twice the accuracy of typical clinical measurement (~10°) and in close agreement with those using cubic spline extrapolation and adaptive neuro-fuzzy inference system (ANFIS) techniques. The adapted technique for predicting the scoliosis deformity progression holds significant promise for clinical applications.