Interactions of Haemophilus parasuis and its LOS with porcine brain microvascular endothelial cells

Haemophilus parasuis is a swine pathogen that causes Gl?sser's disease, which is characterized by polyserositis and meningitis. The pathogenesis of the H. parasuis infection is poorly understood. To cause meningitis, H. parasuis has to cross the blood-brain barrier (BBB) to gain access to the central nervous system (CNS). We recently showed that H. parasuis adheres to and invades porcine brain microvascular endothelial cells (PBMEC). The aim of this study was to evaluate the role of H. parasuis lipooligosaccharide (LOS) in the adhesion to PBMEC and to determine if H. parasuis (and/or its LOS) is able to induce apoptosis and activation of PBMEC. Results showed that adhesion of H. parasuis to PBMEC was partially mediated by LOS. Moreover, H. parasuis induces caspase-3-mediated apoptosis of PBMEC in a time--and dose--dependent manner, but its LOS did not seem to be involved in such a process. Furthermore, H. parasuis and, to a lesser extent, its LOS, was able to induce the release of IL-8 and IL-6 by PBMEC. Field strains of H. parasuis serotypes 4 and 5 induced similar levels of these inflammatory mediators. Our data suggest that H. parasuis uses cellular adhesion, induction of apoptosis and up-regulation of inflammatory mediators as mechanisms to invade the CNS via the BBB, and that LOS would play a certain but limited role in such pathological process.     

Microscopical Study of the Destructuring of Waxy Maize and Smooth Pea Starches by Shear and Heat at Low Hydration

Smooth pea and waxy maize starches were processed under controlled temperature (<160 °C) conditions at low moisture content (30%) using a rheometer with pre-shearing, to simulate extrusion. Specific mechanical energy was measured and remained lower than 10³J/g. Starch modification was analysed at different structural levels by differential scanning calorimetry (DSC), X-ray diffractometry, light microscopy and at the granular level by transmission electron microscopy (TEM). Thermal inputs induced crystallite melting and internal disorganisation of granular structure, whereas mechanical input (SME>10²J/g) induced granule fragmentation. Under shearing and heating, the concomitant destructuring mechanisms led to a higher rate of modification. Fragmentation itself induced only partial loss of crystallinity (<50%). Small fragments finally melted due to a local temperature increase contributed by interparticular friction. Susceptibility to fragmentation was related to the mechanical properties of starch granules associated with their architecture and botanical origin.     

Antifungal activity of Piper guineense of Cameroon

The antifungal activity of the ethanol extract of the seeds of Piper guineense and some of its fractions obtained by column chromatography was investigated using filamentous fungi and yeasts. The results indicated a significant antifungal effect. No toxic effect was observed in mice treated per os with the extract.     

Antifungal activities of five Combretaceae used in Togolese traditional medicine

Five species of Combretaceae growing in Togo were investigated for their antifungal activity against 20 pathogenic fungi (10 yeast and 10 filamentous fungi). The five hydroethanolic extracts of Terminalia glaucescens and Anogeissus leiocarpus appeared to be the most active, their MICs ranging from 0.25 mg/ml to 4 mg/ml. The results confirm the traditional therapeutic properties of these plants.     

3D Reconstruction of the Mouse's Mesonephros

The present work reports on the three-dimensional reconstruction of the segmented mesonephros during the embryonic development of the mouse. With a light microscope and an automatic reconstruction of surfaces, aspects of the mesonephros are described. These surfaces are obtained by using digitized contour lines. A new interpolation method called DSI (Discrete Smooth Interpolation) enables correction of the distortion induced by microtomy in paraffin sections. After a tri-angulation step, this method uses a smoothing algorithm, which implies a spatial redistribution of the vertices of the triangles to correct the rotational and translational misalignment. The use of this 3D program improves the understanding of the development patterns and helps us to appreciate changes in the rebuilt mesonephros. By 10.5 embryonic days, tubules emerge from the Wolffian body and begin their formation, then between 11.5 and 13.5 embryonic days, tortuous mesonephric tubules bound to the Wolffian duct form small curls, which grow and finally unwind. At the same time, mesonephric tubules unbound to the Wolffian duct appear, and on 13.5 embryonic days, the Müllerian duct is visible. After 14.5 embryonic days, the segmented mesonephros keeps its general aspect but decreases in size. At this time, each gonad is provided with both Wolffian and Müllerian ducts. Later, the Wolffian duct differentiates into the definitive male duct system, whilst the Müllerian duct regresses. Conversely, the paramesonephric duct differentiates into the definitive female duct system, whilst the mesonephric duct in turn degenerates. By this time degeneration has begun in the cranial portion of the mesonephros and this process progresses caudally. The spatial organization of the mesonephric tubules and the precise organization of all connections between these elements and the ducts may be well defined. Such approach can allow for a high definition of the normal pattern of mesonephros differentiation.     

Textural Images Analysis of Pasta Protein Networks to Determine Influence of Technological Processes

The structure of pasta is largely governed by the presence of a structured protein network. This work analyzed the protein network textures of various cooked pasta products through textural image analysis. Six different pasta types were investigated: reference pasta made from durum semolina; pasta enriched with gluten proteins from soft wheat flour at 10 and 20%; autoclaved pasta; soft wheat flour pasta; and pasta made from reconstituted flour fractions. Pasta samples were sectioned, and each crosssection consisted of three distinct zones (central, intermediate, and external) based on the state of swelling of starch granules for each pasta product. Digital images of the protein network in each zone were acquired using confocal laser scanning microscopy. Textural image analysis was then performed. Similarities and differences in protein network texture were assessed by principal component, stepwise discriminant, and variance analyses. With the exception of autoclaved pasta, protein network structure differed greatly with the position in the pasta. Furthermore the effect of technological treatments was greatly influenced by the position in pasta. The most significant differences in protein network structure were obtained with the autoclaved and 20% protein-enriched samples.     

Effects of two wheat cultivars on physico-chemical properties of wheat flours and digesta from two broiler chicken lines (D+ and D-) differing in digestion capacity

1. The current experiment is the second part of a study about the effects of wheat quality on digestibility of pelleted diets for broiler chickens. In the first part, it was shown that a hard cultivar resulted in a negative effect on starch digestibility in two divergent lines of chickens (D+ and D-) selected for digestion capacity. The aim of this second part was to investigate the reasons for this negative effect of a hard cultivar (Baltimor) compared to a soft one (Scipion) in D+ and D- lines. 2. Proventriculus pepsin activity and pancreas proteolytic and amylolytic activities were estimated in 4 pools of birds: 'D+ line (Baltimor fed)', 'D+ line (Scipion fed)', 'D- line (Baltimor fed)' and 'D- line (Scipion fed)'. Results suggested the greatest amount of pepsin units per g BW for D+ birds and the lowest amount of pancreas proteolytic units per g BW for D+ birds fed Scipion wheat. Pancreas showed very similar alpha-amylase activities among treatments. 3. In vitro hydrolyses of wheat gluten proteins with proventriculus extracts from pools of D+ and D- birds did not show any differences between hard and soft cultivars, whatever the origin of pools. 4. Pepsin hydrolysis of fine (300 to 425 microm) and coarse (1180 to 1600 microm) fractions from wheat flours (Baltimor or Scipion) showed that the 30 min proteolysis rate was highest for the fine fraction in both cultivars. No difference was observed with extended hydrolysis time. 5. In vitro digestion simulation of whole wheat flours confirmed the results previously obtained in vivo, with a negative effect of hard cultivar on starch digestion rate and no effect on protein digestion. 6. Laser particle size analyses showed that ileum digesta from birds fed with hard wheat cultivar showed the highest proportion of coarse particles. 7. Microscopic analyses of D+ ileum digesta revealed that the concentration of undigested starch granules in the subaleurone area of wheat bran particles was the highest with hard cultivar. 8. The results suggested that physical entrapment of starch granules in coarse particles was a major explanation for decreased starch digestibility values in chickens fed hard wheat diets.     

Wheat endosperm cell walls: Spatial heterogeneity of polysaccharide structure and composition using micro-scale enzymatic fingerprinting and FT-IR microspectroscopy

Micro-scale enzymatic fingerprinting and FT-IR microspectroscopy were used to investigate changes of polysaccharide structure and composition in cell walls from wheat endosperm. These methods were applied to transverse and longitudinal sections of wheat grains harvested at maturity and 270°D. Principal component analysis treatment of the data revealed marked differences in the (1,3)-(1,4)-beta-glucans (BG)/arabinoxylans (AX) ratio and in the structure of AX depending on both cell position in the grain and stage of development. Cell walls close to the germ were enriched in BG in both developing and mature grain. AX in developing grain were characterized by a higher proportion of di-substitution by arabinose as compared to mature grain AX. In addition, AX in mature grains exhibited a different structure depending on prismatic or central origin of cell walls in the grain. These results indicated a high spatial and temporal control of the biosynthesis of AX and BG in wheat endosperm.