Two-photon microscopy : sequential imaging studies in vivo

Microscopists have always desired to look inside various organ tissues to study structure, function and dysfunction of their cellular constituents. In the past, this has frequently required tissue extraction and histological preparation to gain access. Traditional optical microscopy techniques, which use linear (one-photon) absorption processes for contrast generation, are limited to use near the tissue surface (< 80 µm) because at greater depths strong and multiple light scattering blurs the images. Scattering particularly strongly affects signal strength in confocal microscopy, which achieves three-dimensional resolution and optical sectioning with a detection pinhole that rejects all light that appears not to originate from the focus. New optical microscopy techniques have been developed that use nonlinear light-matter interactions to generate signal contrast only within a thin raster-scanned plane. Since its first demonstration over a decade ago, two-photon microscopy has been applied to a variety of imaging tasks and has now become the technique of choice for fluorescence microscopy in thick tissue preparations and in live animals. The gain in resolution over conventional in vivo imaging techniques has been several orders of magnitude. Neuroscientists have used it to measure calcium dynamics deep in brain slices and in live animals, blood flow measurement, neuronal plasticity and to monitor neurodegenerative disease models in brain slices and in live rodents. These types of applications define the most important niche for two-photon microscopy - high-resolution imaging of physiology, morphology and cell-cell interactions in intact tissue. Clearly the biggest advantage of two-photon microscopy is in longitudinal monitoring of rodent models of disease or plasticity over days to weeks. The aim of this article is to discuss some methodological principles, and show some applications of this technique obtained from our laboratory in the area of acute experimental stroke research.peer-reviewe

Vulnerability of white matter to ischemia varies during development

Stroke is the one of the leading causes of mortality and morbidity in developed countries. The central role of injury to white matter in the pathophysiology of stroke has been recognised over the recent years. Stroke can affect a wide range of the population (from the premature infant to the elderly) and therefore the mechanism of injury of central white matter may vary with age. The main aim of this review paper is to shed some light on the difference in maturation of injury to the axon-oligodendrocyte unit following an ischemic insult between different developmental stages. Both components of this unit exhibit varying degrees of susceptibility to ischemia throughout their development. Axons are particularly resistant to ischemia in the neonatal stage. However, they show a marked decreased in tolerance to ischemia during the period of myelination. Late oligodendrocyte progenitor cells (OPC) are the most sensitive type of oligodendrocyte, and their role in periventricular leukomalacia (PVL) is well known. On the other hand, early OPC are particularly resistant to ischemia. Studying the effect of ischemia on white matter in the brain during the different developmental stages will lead to a better understanding of the pathophysiology of white matter injury and hopefully, in the future, to the development of new therapeutic strategies of the various white matter diseases.peer-reviewe

Lateral Habenula contribution in Nicotine addiction : focus on Dopamine, GABA and Serotonin Interactions

Compelling evidence has shown a pivotal role of dopaminergic function in drug addiction. Recently, the Habenula (Hb) has attracted a great deal of attention as another target for nicotine in the brain because of its role in regulating dopamine (DA), gamma-aminobutyric acid (GABA) and serotonin (5-HT) systems. Nicotine acts binding to acetylcholine receptors that are widely distributed in the brain. Interestingly, the receptor subtypes that mediate nicotine withdrawal responses are highly expressed in the Hb. Moreover, the block of habenular nicotinic receptors in animals chronically treated with nicotine enhances withdrawal responses once nicotine is discontinued. Furthermore, it has been shown how a high dose of nicotine can cause massive degeneration almost exclusively in the medial habenula (MHb) and its output tract, the fasciculus retroflexus. Thus, symptoms associated with nicotine withdrawal may be caused by dysfunctions of the Hb output. Therefore, Hb might be of fundamental importance in the expression of nicotine reinforcing properties and withdrawal. Here, we will focus on the role of the lateral habenula (LHb) on nicotine modulation of DA function and we will evaluate LHb interaction with the rostromedial tegmental nucleus (RMTg), a GABAergic area, and the serotonergic raphé nuclei. Furthermore, as LHb has high density expression of 5-HT2C receptors, these subtypes might be important in the control of its neuronal activity and output to the midbrain monoaminergic and GABAergic systems.peer-reviewe

Relations between organic and terrigenous matter deposition in Holocene fluvial sediments: heterodox observations and conclusions.

International audienceAccumulation of organic matter in fens of fluvial valleys is often related to a low terrigenous matter delivery and to palaeoenvironmental conditions inducing low mechanical erosion. These assumptions come from the interpretation of contents in organic (MO) and mineral (MM) matters in sediments, expressed in percents, and then exactly anticorrelated. Calculation of mass accumulation rates of MO (TaMO) and MM (TaMM), expressed in g m−2 yr−1, shows that TaMO and TaMM generally are not anticorrelated and that high values of TaMO and TaMM could appear simultaneously. That expression of MO and MM accumulation makes it possible to precise the climatic and human impact on sedimentation

Storage and release of fossil organic carbon related to weathering of sedimentary rocks

International audienceThe biogeochemical carbon cycle, which plays an undeniable role in global climate change, is defined both by the size of carbon reservoirs (such as the atmosphere, biomass, soil and bedrock) and the exchange between them of various mineral and organic carbon forms. Among these carbon forms, fossil organic carbon (FOC) (i.e., the ancient organic matter stored in sedimentary rocks) is widely observed in modern environments but is not included in the supergene carbon budget. Using a digitized map of the world and an existing model of CO2 consumption associated with rock weathering, we establish the global distribution of FOC stored in the first meter of sedimentary rocks and a first estimation of annual FOC delivery to the modern environment resulting from chemical weathering of these rocks. Results are given for the world's 40 major river basins and extended to the entire continental surface. With a mean value of 1100 109 t, mainly controlled by shale distribution, the global FOC stock is significant and comparable to that of soil organic carbon (1500 109 t). The annual chemical delivery of FOC, estimated at 43 106 t yr− 1 and controlled by the areal distribution of shales and runoff, is of the same order of magnitude as the FOC output flux to oceans. Chemical weathering of bedrock within the Amazon basin produces one-quarter of the total global flux of FOC derived from chemical weathering, and thus is expected to govern FOC release on a global scale. These results raise important questions concerning the role of FOC in the modern carbon cycle as well as the origin and the budget of carbon in soils and river

Estimation of the annual organic carbon yield related to carbonated rocks chemical weathering : implications for the global organic carbon cycle understanding.

The aim of this paper is to propose an initial estimation of the annual organic matter yield induced by chemical weathering of carbonates and shales, considering their global surface at outcrop and their organic matter content. The calculation also uses data on river fluxes resulting from carbonate rocks and shales weathering in major world watersheds, published by numerous authors. The results obtained from the studied watersheds have then been extrapolated to a global scale. Despite rather large uncertainty to such an approach, the calculated value of ca. 0.1 Gt implies that the annual organic carbon yield related to carbonates and shales chemical weathering might be a non-negligible component of the global carbon cycle. The individual contributions of different watersheds necessarily depend on the organic matter content of altered rocks. They are also obviously controlled by climatic parameters. The calculated yields do not constitute a direct supply to soils and rivers because of mineralisation when organic carbon is brought in contact with the atmosphere. Even so, the release of fossil organic matter would have implications for the global carbon cycle through the efficiency of the global chemical weathering as a carbon sink. Whatever the chosen hypothesis, the results of this study suggest that the recycled organic yield is a neglected component in the global organic carbon cycle assessment. Because it exists and, in addition, because it might represent a non-negligible carbon pool, fossil organic carbon deserves to be taken into account for a better evaluation of the organic stocks in soils and rivers presently only based on climatic data and current vegetal production

The central role of aquaporins in the pathophysiology of ischemic stroke

Stroke is a complex and devastating neurological condition with limited treatment options. Brain edema is a serious complication of stroke. Early edema formation can significantly contribute to infarct formation and thus represents a promising target. Aquaporin (AQP) water channels contribute to water homeostasis by regulating water transport and are implicated in several disease pathways. At least 7 AQP subtypes have been identified in the rodent brain and the use of transgenic mice has greatly aided our understanding of their functions. AQP4, the most abundant channel in the brain, is up-regulated around the peri-infarct border in transient cerebral ischemia and AQP4 knockout mice demonstrate significantly reduced cerebral edema and improved neurological outcome. In models of vasogenic edema, brain swelling is more pronounced in AQP4-null mice than wild-type providing strong evidence of the dual role of AQP4 in the formation and resolution of both vasogenic and cytotoxic edema. AQP4 is co-localized with inwardly rectifying K+-channels (Kir4.1) and glial K+ uptake is attenuated in AQP4 knockout mice compared to wild-type, indicating some form of functional interaction. AQP4-null mice also exhibit a reduction in calcium signaling, suggesting that this channel may also be involved in triggering pathological downstream signaling events. Associations with the gap junction protein Cx43 possibly recapitulate its role in edema dissipation within the astroglial syncytium. Other roles ascribed to AQP4 include facilitation of astrocyte migration, glial scar formation, modulation of inflammation and signaling functions. Treatment of ischemic cerebral edema is based on the various mechanisms in which fluid content in different brain compartments can be modified. The identification of modulators and inhibitors of AQP4 offer new therapeutic avenues in the hope of reducing the extent of morbidity and mortality in stroke.peer-reviewe

Past Holocene detritism quantification and modeling from lacustrine archives in order to deconvoluate human-climate interactions on natural ecosystem over long time-scale

International audienceWater budget is one of the main challenges to paleoclimate researchers in relation to present-day global warming and its consequences for human societies

Using Rock-Eval 6 pyrolysis for tracking fossil organic carbon in modern environments: implications for the roles of erosion and weathering.

This work relates to the debate on the fossil organic carbon (FOC) input in modern environments and its possible implication for the carbon cycle, and suggests the use of Rock-Eval 6 pyrolysis as a relevant tool for tracking FOC in such environments. Considering that such a delivery is mainly due to supergene processes affecting the continental surface, we studied organic matter in different reservoirs such as bedrocks, alterites, soils and rivers in two experimental catchments at Draix (Alpes de Haute Provence, France). Samples were subjected to geochemical (Rock-Eval 6 pyrolysis) investigations and artificial bacterial degradations. After comparing the geochemical fingerprint of samples, geochemical markers of FOC were defined and tracked in the different reservoirs. Our results confirm the contribution of FOC in modern soils and rivers and display the various influences of weathering and erosional processes on the fate of FOC during its exchange between these pools. In addition, the contrasting behaviour of these markers upon the supergene processes has also highlighted the refractory or labile characters of the fossil organic matter (FOM). Bedrock to river fluxes, controlled by gully erosion, are characterized by a qualitative and quantitative preservation of FOM. Bedrock to alterite fluxes, governed by chemical weathering, are characterized by FOC mineralization without qualitative changes in deeper alterites. Alterite to soils fluxes, controlled by (bio)chemical weathering, are characterized by strong FOC mineralization and qualitative changes of FOM. Thus weathering and erosional processes induce different FOM evolution and affect the fate of FOC towards the global carbon cycle. In this study, gully erosion would involve maintenance of an ancient sink for the global carbon cycle, while (bio)chemical processes provide a source of CO2. Finally, this study suggests that Rock-Eval 6 pyrolysis can be considered as a relevant tool for tracking FOC in modern environments

Screening for Tuberculosis in Health Care Workers. Experience in an Italian Teaching Hospital

Health care workers (HCW) are particularly at risk of acquiring tuberculosis (TB), even in countries with low TB incidence. Therefore, TB screening in HCW is a useful prevention strategy in countries with both low and high TB incidence. Tuberculin skin test (TST) is widely used although it suffers of low specificity; on the contrary, the in vitro enzyme immunoassay tests (IGRA) show superior specificity and sensitivity but are more expensive. The present study reports the results of a three-year TB surveillance among HCW in a large teaching hospital in Rome, using TST (by standard Mantoux technique) and IGRA (by QuantiFERON-TB) as first- and second-level screening tests, respectively. Out of 2290 HCW enrolled, 141 (6.1%) had a positive TST; among them, 99 (70.2%) underwent the IGRA and 16 tested positive (16.1%). The frequency of HCW tested positive for TB seems not far from other experiences in low incidence countries. Our results confirm the higher specificity of IGRA, but, due to its higher cost, TST can be considered a good first level screening test, whose positive results should be further confirmed by IGRA before the patients undergo X-ray diagnosis and/or chemotherapy