In utero alcohol exposure impairs vessel-associated positioning and differentiation of oligodendrocytes in the developing neocortex.

Prenatal alcohol exposure (PAE) is a major cause of nongenetic mental retardation and can lead to fetal alcohol syndrome (FAS), the most severe manifestation of fetal alcohol spectrum disorder (FASD). FASD infants present behavioral disabilities resulting from neurodevelopmental defects. Both grey and white matter lesions have been characterized and are associated with apoptotic death and/or ectopic migration profiles. In the last decade, it was shown that PAE impairs brain angiogenesis, and the radial organization of cortical microvessels is lost. Concurrently, several studies have reported that tangential migration of oligodendrocyte precursors (OPCs) originating from ganglionic eminences is vascular associated. Because numerous migrating oligodendrocytes enter the developing neocortex, the present study aimed to determine whether migrating OPCs interacted with radial cortical microvessels and whether alcohol-induced vascular impairments were associated with altered positioning and differentiation of cortical oligodendrocytes. Using a 3D morphometric analysis, the results revealed that in both human and mouse cortices, 15 to 40% of Olig2-positive cells were in close association with radial cortical microvessels, respectively. Despite perinatal vascular disorganization, PAE did not modify the vessel association of Olig2-positive cells but impaired their positioning between deep and superficial cortical layers. At the molecular level, PAE markedly but transiently reduced the expression of CNPase and MBP, two differentiation markers of immature and mature oligodendrocytes. In particular, PAE inverted their distribution profiles in cortical layers V and VI and reduced the thickness of the myelin sheath of efferent axons. These perinatal oligo-vascular defects were associated with motor disabilities that persisted in adults. Altogether, the present study provides the first evidence that Olig2-positive cells entering the neocortex are associated with radial microvessels. PAE disorganized the cortical microvasculature and delayed the positioning and differentiation of oligodendrocytes. Although most of these oligovascular defects occurred in perinatal life, the offspring developed long-term motor troubles. Altogether, these data suggest that alcohol-induced oligo-vascular impairments contribute to the neurodevelopmental issues described in FASD.

A drop does not fall in a straight line: a rationale for the width of stalagmites.

Drops loaded in calcium ions detach from stalactites and impact the underlying stalagmites, thereby allowing these latter to grow through calcite precipitation. Nevertheless, little is known about the influence of the drop free fall and splash dynamics on stalagmite shape and width. Through high-speed imaging of impacting drops on stalagmites from several caves, we observed that the impact point position of the drops is scattered, sometimes over several centimetres. We show that this dispersal has no external cause and must, therefore, be self-induced. Using a Langevin-like equation, we then propose a prediction of the impact point dispersal as a function of the falling height travelled by the drops. We finally show that measured stalagmite widths are correlated to the dispersal in the impact point position of the drop.

Electron microscopy approach for the visualization of the epithelial and endothelial glycocalyx.

This study presents a methodological approach for the visualization of the glycocalyx by electron microscopy. The glycocalyx is a three dimensional network mainly composed of glycolipids, glycoproteins and proteoglycans associated with the plasma membrane. Since less than a decade, the epithelial and endothelial glycocalyx proved to play an important role in physiology and pathology, increasing its research interest especially in vascular functions. Therefore, visualization of the glycocalyx requires reliable techniques and its preservation remains challenging due to its fragile and dynamic organization, which is highly sensitive to the different process steps for electron microscopy sampling. In this study, chemical fixation was performed by perfusion as a good alternative to conventional fixation. Additional lanthanum nitrate in the fixative enhances staining of the glycocalyx in transmission electron microscopy bright field and improves its visualization by detecting the elastic scattered electrons, thus providing a chemical contrast.

Conservation of prehistoric caves and stability of their inner climate: lessons from Chauvet and other French caves.

In the last 150 years, some prehistoric painted caves suffered irreversible degradations due to misperception of conservation issues and subsequent mismanagement. These sites presented naturally an exceptional stability of their internal climate allowing conservation in situ of outstanding fragile remains, some for nearly 40,000 years. This is for a large part due to exchanges of air, CO2, heat and water with the karstic system in which these caves are included. We introduce the concept of underground confinement, based on the stability of the inner cave climate parameters, especially its temperature. Confined caves present the best conservative properties. It is emphasized that this confined state implies slow exchanges with the surrounding karst and that a stable cave cannot be viewed as a closed system. This is illustrated on four case studies of French caves of various confinement states evidenced by long term continuous monitoring and on strategies to improve their conservation properties. The Chauvet cave presents optimal conservation properties. It is wholly confined as shown by the stability of its internal parameters since its discovery in 1994. In Marsoulas cave, archeological works removed the entrance scree and let a strong opening situation of the decorated zone. Remediation is expected by adding a buffer structure at the entrance. In Pech Merle tourist cave, recurrent painting fading was related to natural seasonal drying of walls. Improvement of the cave closure system restored a confined state insuring optimal visibility of the paintings. In Gargas tourist cave, optimization of closures, lighting system and number of visitors, allowed it to gradually reach a semi-confined state that improved the conservation properties. Conclusions are drawn on the characterization of confinement state of caves and on the ways to improve their conservation properties by restoring their initial regulation mechanisms and to avoid threats to their stability.

[Secondary opacification of hydrophilic acrylic intraocular lenses: report of two cases]. / Opacification secondaire d'implants intraoculaires acryliques hydrophiles : à propos de deux cas.

PURPOSE: To describe two cases of secondary opacification of hydrophilic acrylic IOLs after vitreoretinal surgery. METHODS: Analysis of IOL deposits with anterior segment optical coherence tomography (OCT Visante(®)) and high-frequency ultrasound biomicroscopy (CineScan HF, Quantel Medical(®)) was carried out preoperatively. The explanted IOLs were analyzed with optical and electron microscopy, and energy dispersive spectrometry (EDS). RESULTS: In both patients, the deposits were located on the surface and within the implant and were composed of calcium phosphate crystals. Vitreoretinal surgery performed a few months prior to the secondary opacification was identified as a risk factor. CONCLUSION: When a patient presents with a secondary opacification of the IOL involving the visual axis, explantation is sometimes necessary, fortunately with typically good functional recovery in the postoperative period. While this complication is rare and the exact pathophysiology poorly understood, it must be considered in a pseudophakic patient with an unexplained decrease in visual acuity who has undergone more than one intraocular procedure.

Precise dating of Dansgaard-Oeschger climate oscillations in western Europe from stalagmite data.

The signature of Dansgaard-Oeschger events--millennial-scale abrupt climate oscillations during the last glacial period--is well established in ice cores and marine records. But the effects of such events in continental settings are not as clear, and their absolute chronology is uncertain beyond the limit of (14)C dating and annual layer counting for marine records and ice cores, respectively. Here we present carbon and oxygen isotope records from a stalagmite collected in southwest France which have been precisely dated using 234U/230Th ratios. We find rapid climate oscillations coincident with the established Dansgaard-Oeschger events between 83,000 and 32,000 years ago in both isotope records. The oxygen isotope signature is similar to a record from Soreq cave, Israel, and deep-sea records, indicating the large spatial scale of the climate oscillations. The signal in the carbon isotopes gives evidence of drastic and rapid vegetation changes in western Europe, an important site in human cultural evolution. We also find evidence for a long phase of extremely cold climate in southwest France between 61.2 +/- 0.6 and 67.4 +/- 0.9 kyr ago.

Transmucosal transport of phytohemagglutinin, concanavalin A and alpha-lactalbumin across rabbit ileum in vitro.

The transmucosal passage of alpha-lactalbumin (alpha-La), phytohemagglutinin (PHA), and concanavalin A (Con A) (1 mg/ml) was measured in the rabbit ileum mounted in the Ussing chamber, with and without 10(-2) M glucose or galactose. The transport of the radiolabelled proteins was assessed by radioisotopic determination, high pressure liquid chromatography (HPLC) and enzyme linked immunosorbent assay (ELISA). In the absence of galactose, the transmucosal transport was significantly higher for PHA (4.1 +/- 1.8 micrograms/h.cm2 mean +/- SE) than for alpha-La (2.9 +/- 1.2) and very low for Con A (0.6 +/- 0.5). HPLC analysis of the transported material revealed differential processing of the proteins. ELISA indicated that 3 percent of radiolabelled alpha-La that crossed the epithelium was in an immunoreactive form, whereas no immunoreactive forms of PHA and Con A were detected. The uptake or binding by the tissue was identical for PHA and Con A (7.8 +/- 2.9 and 5.8 +/- 2.8 micrograms/cm2, respectively), and significantly lower for alpha-La (1.5 +/- 0.31). 10(-2) M galactose did not modify the uptake or binding of alpha-La and Con A, but significantly decreased that of PHA to a level that was not significantly different from that of alpha-La. The present results indicate that the initial uptake of the proteins is most likely dependent upon their interactions with the luminal side of the epithelium. After uptake, the proteins are subjected to intracellular processing which also appeared differential. Thus, protein transport depends on the properties both of the compartment crossed (Glycocalyx, brush-border membrane, cytoplasm, basolateral membrane), and of the protein.

Transport of beta-lactoglobulin across rabbit ileum in vitro.

Intestinal transepithelial transport constitutes a major limiting step in the transfer of food protein antigens to the blood. This transport was studied in isolated rabbit ileum in Ussing chamber in vitro for the milk protein antigen beta-lactoglobulin (beta-Lg). The transepithelial passage of beta-Lg was measured by enzyme-linked immunosorbent assay (ELISA) and radiolabeled protein transfer and compared with that of the nonmetabolizable marker polyethylene glycol (PEG)-4000. When 1 mg/ml of beta-[14C]Lg or [3H]PEG was added to the mucosal side of the tissue, the total uptake, measured as the transfer of radiolabeled material across the ileum, was significantly higher for beta-Lg than for PEG (5.46 +/- 1.75 vs. 1.43 +/- 0.26 micrograms.h-1.cm-2). Measured by ELISA, 6-9% of the total amount of beta-Lg transported was absorbed in an intact antigenic form. This transport of intact beta-Lg was inhibited by the metabolic inhibitors 50 mM 2-deoxyglucose and 1 mM azide added simultaneously, was reduced by the microtubule assembly inhibitor 0.05 mM colchicine, and was enhanced by 20 mM ammonia, which inhibits lysosomal proteolytic activity. These results indicate that beta-Lg is efficiently absorbed by the intestinal mucosa of adult animals, partly in intact antigenic form and that beta-Lg transport is probably transcellular, as observed for other proteins. The finding that beta-Lg is absorbed in intact antigenic form agrees with other reports implying that beta-Lg is the main factor responsible for milk protein immunoreactivity and intolerance.

Permeability of milk protein antigens across the intestinal epithelium in vitro.

Degradations by proteolytic enzymes and intestinal epithelial permeability represent two major drawbacks to the transfer of food protein antigens to blood. These steps were studied in vitro for the milk protein antigens beta-lactoglobulin (beta-Lg), alpha-Lactalbumin (alpha-La) and beta-casein (beta-cas). Pepsin-trypsin hydrolysis and permeability in isolated rabbit ileum in Ussing chamber were suited by ELISA and radiolabelled-protein measurement. Pepsin-trypsin hydrolysis showed an increasing resistance in the order beta-cas less than alpha-La less than beta-Lg. The rate of absorption of the antigenic proteins by isolated rabbit ileum was in the same order, and the rate of absorption of the whole proteins (degraded and antigenic forms) was significantly higher for beta-Lg than for alpha-La and beta-cas. These results suggest a selective intestinal permeability for milk protein antigens. This selectivity is probably important in the mechanism of food protein sensitization via the oral route.