Review: leucocyte-endothelial cell crosstalk at the blood-brain barrier: a prerequisite for successful immune cell entry to the brain

Leucocyte migration into the central nervous system is a key stage in the development of multiple sclerosis. While much has been learnt regarding the sequential steps of leucocyte capture, adhesion and migration across the vasculature, the molecular basis of leucocyte extravasation is only just being unravelled. It is now recognized that bidirectional crosstalk between the immune cell and endothelium is an essential element in mediating diapedesis during both normal immune surveillance and under inflammatory conditions. The induction of various signalling networks, through engagement of cell surface molecules such as integrins on the leucocyte and immunoglobulin superfamily cell adhesion molecules on the endothelial cell, play a major role in determining the pattern and route of leucocyte emigration. In this review we discuss the extent of our knowledge regarding leucocyte migration across the blood-brain barrier and in particular the endothelial cell signalling pathways contributing to this process.

Going against the tide--how encephalitogenic T cells breach the blood-brain barrier

During multiple sclerosis or its animal model, experimental autoimmune encephalomyelitis, circulating immune cells enter the central nervous system (CNS) causing neuroinflammation. Extravasation from the blood circulation across the vessel wall occurs through a multistep process regulated by adhesion and signal transducing molecules on the immune cells and on the endothelium. Since the CNS is shielded by the highly specialized blood-brain barrier (BBB), immune cell extravasation into the CNS requires breaching this particularly tight endothelial border. Consequently, travelling into the CNS demands unique adaptations which account for the extreme tightness of the BBB. Modern imaging tools have shown that after arresting on BBB endothelium, in vivo or in vitro encephalitogenic effector/memory T cells crawl for long distances, possibly exceeding 150 µm along the surface of the BBB endothelium before rapidly crossing the BBB. Interestingly, in addition to the distance of crawling, the preferred direction of crawling against the flow is unique for T cell crawling on the luminal surface of CNS microvessels. In this review, we will summarize the cellular and molecular mechanisms involved in the unique T cell behavior that is obviously required for finding a site permissive for diapedesis across the unique vascular bed of the BBB.

Surface uplift and climate change - the geomorphic evolution of the western escarpment of the Andes of northern Chile between the Miocene and present

The Western Escarpment of the Andes at 18.30°S (Arica area, northern Chile) is a classical example for a transient state in landscape evolution. This part of the Andes is characterized by the presence of >10,000 km2 plains that formed between the Miocene and the present, and >1500 m deeply incised valleys. Although processes in these valleys scale the rates of landscape evolution, determinations of ages of incision, and more importantly, interpretations of possible controls on valley formation have been controversial. This paper uses morphometric data and observations, stratigraphic information, and estimates of sediment yields for the time interval between ca. 7.5 Ma and present to illustrate that the formation of these valleys was driven by two probably unrelated components. The first component is a phase of base-level lowering with magnitudes of∼300–500 m in the Coastal Cordillera. This period of base-level change in the Arica area, that started at ca. 7.5 Ma according to stratigraphic data, caused the trunk streams to dissect headward into the plains. The headward erosion interpretation is based on the presence of well-defined knickzones in stream profiles and the decrease in valley widths from the coast toward these knickzones. The second component is a change in paleoclimate. This interpretation is based on (1) the increase in the size of the largest alluvial boulders (from dm to m scale) with distal sources during the last 7.5 m.y., and (2) the calculated increase in minimum fluvial incision rates of ∼0.2 mm/yr between ca. 7.5 Ma and 3 Ma to ∼0.3 mm/yr subsequently. These trends suggest an increase in effective water discharge for systems sourced in the Western Cordillera (distal source). During the same time, however, valleys with headwaters in the coastal region (local source) lack any evidence of fluvial incision. This implies that the Coastal Cordillera became hyperarid sometime after 7.5 Ma. Furthermore, between 7.5 Ma and present, the sediment yields have been consistently higher in the catchments with distal sources (∼15 m/m.y.) than in the headwaters of rivers with local sources (<7 m/m.y.). The positive correlation between sediment yields and the altitude of the headwaters (distal versus local sources) seems to reflect the effect of orographic precipitation on surface erosion. It appears that base-level change in the coastal region, in combination with an increase in the orographic effect of precipitation, has controlled the topographic evolution of the northern Chilean Andes.

Effect of two bioabsorbable barrier membranes on bone regeneration of standardized defects in calvarial bone: a comparative histomorphometric study in pigs

BACKGROUND: The effect of two different bioabsorbable collagen membranes on bone regeneration was assessed in standardized, membrane-protected calvarial defects in pigs. METHODS: Two standardized defect types (6 x 6 x 6 mm and 9 x 9 x 9 mm) were produced in the calvaria of pigs: empty defects without a membrane (group 1; eight defects per size); defects filled with deproteinized bovine bone mineral (DBBM) without a membrane (group 2; eight defects per size); defects filled with DBBM and covered by a collagen membrane (group 3; eight defects per size); and defects filled with DBBM and covered by a cross-linked collagen membrane (CCM) (group 4; eight defects per size). Sacrifice took place 16 weeks after surgery, and the following parameters were analyzed: descriptive histology; semiquantitative histology (SQH), assessing bone regeneration in the whole defect area; and histomorphometric analysis of the percentage of bone and DBBM in the regenerated area at three different depth levels of the defect. RESULTS: Using SQH, both membrane types resulted in significantly better bone regeneration compared to groups 1 and 2, irrespective of the defect size (P <0.005), with no difference between the two membranes. In the histomorphometric analysis, the layer immediately below the surface exhibited a significantly higher percentage of bone in groups 3 (27%) and 4 (36%) versus the two other groups for the 9 x 9 x 9-mm defects. No such differences were apparent for the 6 x 6 x 6-mm defects or the other two depth levels (bottom and middle layer) for either defect size. CONCLUSIONS: The two collagen membranes tested significantly enhanced bone regeneration, especially in the superficial level of the calvarial bone defects. The prototype CCM did not provide any further advantage in the present animal model.

Effects of decontamination and implant surface characteristics on re-osseointegration following treatment of peri-implantitis

BACKGROUND: Although considerable bone fill may occur following treatment of peri-implantitis, re-osseointegration appears to be limited and unpredictable. Objectives: To evaluate the effects of various decontamination techniques and implant surface configurations on re-osseointegration of contaminated dental implants. MATERIAL AND METHODS: Three months after tooth extraction, implants consisting of a basal part and an exchangeable intraosseous implant cylinder (EIIC) were placed in the mandibles of dogs. The EIIC was machined (M), sandblasted and acid-etched (SLA), or titanium plasma sprayed (TPS). Ligature-induced peri-implantitis was initiated 8 weeks post-implantation and lasted until bone loss reached the junction of the two implant parts. Three treatment modalities were applied: (T1) the EIIC was exchanged for a pristine EIIC; (T2) the EIIC was sprayed in situ with saline; and (T3) the EIIC was removed, cleansed outside the mouth by spraying with saline, steam-sterilized, and remounted. A collagen barrier was placed over each fixture, and 3 months later, samples were processed for histology and histomorphometry. RESULTS: T2 revealed the highest bone-to-implant contact (BIC) level (significantly better than T1 and T3). T2 also yielded the highest bone crest level (significantly better than T1), followed by T3 (significantly better than T1). SLA showed the highest BIC level (significantly better than M), followed by TPS. There were no statistically significant differences in bone crest height between implant types. CONCLUSIONS: Both SLA implants and in situ cleansing resulted in the best re-osseointegration and bone fill of previously contaminated implants.

Loss of astrocyte polarity marks blood-brain barrier impairment during experimental autoimmune encephalomyelitis

In multiple sclerosis (MS), and its animal model experimental autoimmune encephalomyelitis (EAE), dysfunction of the blood-brain barrier (BBB) leads to edema formation within the central nervous system. The molecular mechanisms of edema formation in EAE/MS are poorly understood. We hypothesized that edema formation is due to imbalanced water transport across the BBB caused by a disturbed crosstalk between BBB endothelium and astrocytes. Here, we demonstrate at the light microscopic and ultrastructural level, the loss of polarized localization of the water channel protein aquaporin-4 (AQP4) in astrocytic endfeet surrounding microvessels during EAE. AQP4 was found to be redistributed over the entire astrocytic cell surface and lost its arrangement in orthogonal arrays of intramembranous particles as seen in the freeze-fracture replica. In addition, immunostaining for the astrocytic extracellular matrix receptor beta-dystroglycan disappeared from astroglial membranes in the vicinity of inflammatory cuffs, whereas immunostaining for the dystroglycan ligands agrin and laminin in the perivascular basement membrane remained unchanged. Our data suggest that during EAE, loss of beta-dystroglycan-mediated astrocyte foot process anchoring to the basement membrane leads to loss of polarized AQP4 localization in astrocytic endfeet, and thus to edema formation in EAE.

A Closed-Form, Consistent and Robust Solution to Uncalibrated Photometric Stereo Via Local Diffuse Reflectance Maxima

Images of an object under different illumination are known to provide strong cues about the object surface. A mathematical formalization of how to recover the normal map of such a surface leads to the so-called uncalibrated photometric stereo problem. In the simplest instance, this problem can be reduced to the task of identifying only three parameters: the so-called generalized bas-relief (GBR) ambiguity. The challenge is to find additional general assumptions about the object, that identify these parameters uniquely. Current approaches are not consistent, i.e., they provide different solutions when run multiple times on the same data. To address this limitation, we propose exploiting local diffuse reflectance (LDR) maxima, i.e., points in the scene where the normal vector is parallel to the illumination direction (see Fig. 1). We demonstrate several noteworthy properties of these maxima: a closed-form solution, computational efficiency and GBR consistency. An LDR maximum yields a simple closed-form solution corresponding to a semi-circle in the GBR parameters space (see Fig. 2); because as few as two diffuse maxima in different images identify a unique solution, the identification of the GBR parameters can be achieved very efficiently; finally, the algorithm is consistent as it always returns the same solution given the same data. Our algorithm is also remarkably robust: It can obtain an accurate estimate of the GBR parameters even with extremely high levels of outliers in the detected maxima (up to 80 % of the observations). The method is validated on real data and achieves state-of-the-art results.

Soluble FLT1 binds lipid microdomains in podocytes to control cell morphology and glomerular barrier function.

Vascular endothelial growth factor and its receptors, FLK1/KDR and FLT1, are key regulators of angiogenesis. Unlike FLK1/KDR, the role of FLT1 has remained elusive. FLT1 is produced as soluble (sFLT1) and full-length isoforms. Here, we show that pericytes from multiple tissues produce sFLT1. To define the biologic role of sFLT1, we chose the glomerular microvasculature as a model system. Deletion of Flt1 from specialized glomerular pericytes, known as podocytes, causes reorganization of their cytoskeleton with massive proteinuria and kidney failure, characteristic features of nephrotic syndrome in humans. The kinase-deficient allele of Flt1 rescues this phenotype, demonstrating dispensability of the full-length isoform. Using cell imaging, proteomics, and lipidomics, we show that sFLT1 binds to the glycosphingolipid GM3 in lipid rafts on the surface of podocytes, promoting adhesion and rapid actin reorganization. sFLT1 also regulates pericyte function in vessels outside of the kidney. Our findings demonstrate an autocrine function for sFLT1 to control pericyte behavior.

Continental degassing of 4He by surficial discharge of deep groundwater

Radiogenic He is produced by the decay of uranium and thorium in the Earth’s mantle and crust. From here, it is degassed to the atmosphere and eventually escapes to space. Assuming that all of the 4He produced is degassed, about 70% of the total He degassed from Earth comes from the continental crust. However, the outgoing flux of crustal He has not been directly measured at the Earth’s surface and the migration pathways are poorly understood. Here we present measurements of helium isotopes and the long-lived cosmogenic radio-isotope Kr in the deep, continental-scale Guarani aquifer in Brazil and show that crustal He reaches the atmosphere primarily by the surficial discharge of deep groundwater. We estimate that He in Guarani groundwater discharge accounts for about 20% of the assumed global flux from continental crust, and that other large aquifers may account for about 33%. Old groundwater ages suggest that He in the Guarani aquifer accumulates over half- to one-millionyear timescales. We conclude that He degassing from the continents is regulated by groundwater discharge, rather than episodic tectonic events, and suggest that the assumed steady state between crustal production and degassing of He, and its resulting atmospheric residence time, should be re-examined.

On-Surface Synthesis of BN-Substituted Heteroaromatic Networks

We report on the bottom-up fabrication of BN-substituted heteroaromatic networks achieved by surface-assisted polymerization and subsequent cyclodehydrogenation of specifically designed BN-substituted precursor monomers based on a borazine core structural element. To get insight into the cyclodehydrogenation pathway and the influence of molecular flexibility on network quality, two closely related precursor monomers with different degrees of internal cyclodehydrogenation have been employed. Scanning tunneling microscopy shows that, for both monomers, surface-assisted cyclodehydrogenation allows for complete monomer cyclization and the formation of covalently interlinked BN-substituted polyaromatic hydrocarbon networks on the Ag(111) surface. In agreement with experimental observations, density functional theory calculations reveal a significantly lower energy barrier for the cyclodehydrogenation of the conformationally more rigid precursor monomer, which is also reflected in a higher degree of long-range order of the obtained heteroaromatic network. Our proof-of-concept study will allow for the fabrication of atomically precise substitution patterns within BNC heterostructures.

Narrow dust jets in a diffuse gas coma: a natural product of small active regions on comets

Comets often display narrow dust jets but more diffuse gas comae when their eccentric orbits bring them into the inner solar system and sunlight sublimates the ice on the nucleus. Comets are also understood to have one or more active areas covering only a fraction of the total surface active with sublimating volatile ices. Calculations of the gas and dust distribution from a small active area on a comet’s nucleus show that as the gas moves out radially into the vacuum of space it expands tangentially, filling much of the hemisphere centered on the active region. The dust dragged by the gas remains more concentrated over the active area. This explains some puzzling appearances of comets having collimated dust jets but more diffuse gaseous atmospheres. Our test case is 67P/Churyumov–Gerasimenko, the Rosetta mission target comet, whose activity is dominated by a single area covering only 4% of its surface.

Paleohydraulic reconstruction of a 40 kyr-old terrace sequence implies that water discharge was larger than today

The evolution of landscapes crucially depends on the climate history. This is particularly evident in South America where landscape responses to orbital climate shifts have been well documented. However, while most studies have focused on inferring temperature variations from paleoclimate proxy data, estimates of water budget changes have been complicated because of a lack of adequate physical information. Here, we present a methodology and related results, which allowed us to extract water discharge values from the sedimentary record of the 40 Ka-old fluvial terrace deposits in the Pisco valley, western Peru. In particular, this valley hosts a Quaternary cut-and-fill succession that we used, in combination with beryllium-10 (10Be)-based sediment flux, gauging records, channel geometries and grain size measurements, to quantitatively assess sediment and water discharge values c. 40 Ka ago in relation to present-day conditions. We compare these discharge estimates to the discharge regime of the modern Pisco River and find that the water discharge of the paleo-Pisco River, during the Minchin pluvial period c. 40 Ka ago, was c. 7–8 times greater than the modern Pisco River if considering the mean and the maximum water discharge. In addition, the calculations show that inferred water discharge estimates are mainly dependent on channel gradients and grain size values, and to a lesser extent on channel width measures. Finally, we found that the c. 40 Ka-old Minchin terrace material was poorer sorted than the modern deposits, which might reflect that sediment transport during the past period was characterized by a larger divergence from equal mobility compared to the modern situation. In summary, the differences in grain size distribution and inferred water discharge estimates between the modern and the paleo-Pisco River suggests that the 40 Ka-old Minchin period was characterized by a wetter climate and more powerful flood events.

Real-Time In Vivo Characterization of Primary Liver Tumors With Diffuse Optical Spectroscopy During Percutaneous Needle Interventions: Feasibility Study in Woodchucks

OBJECTIVE This study presents the first in vivo real-time optical tissue characterization during image-guided percutaneous intervention using near-infrared diffuse optical spectroscopy sensing at the tip of a needle. The goal of this study was to indicate transition boundaries from healthy tissue to tumors, namely, hepatic carcinoma, based on the real-time feedback derived from the optical measurements. MATERIALS AND METHODS Five woodchucks with hepatic carcinoma were used for this study. The woodchucks were imaged with contrast-enhanced cone beam computed tomography with a flat panel detector C-arm system to visualize the carcinoma in the liver. In each animal, 3 insertions were performed, starting from the skin surface toward the hepatic carcinoma under image guidance. In 2 woodchucks, each end point of the insertion was confirmed with pathologic examination of a biopsy sample. While advancing the needle in the animals under image guidance such as fluoroscopy overlaid with cone beam computed tomography slice and ultrasound, optical spectra were acquired at the distal end of the needles. Optical tissue characterization was determined by translating the acquired optical spectra into clinical parameters such as blood, water, lipid, and bile fractions; tissue oxygenation levels; and scattering amplitude related to tissue density. The Kruskal-Wallis test was used to study the difference in the derived clinical parameters from the measurements performed within the healthy tissue and the hepatic carcinoma. Kurtoses were calculated to assess the dispersion of these parameters within the healthy and carcinoma tissues. RESULTS Blood and lipid volume fractions as well as tissue oxygenation and reduced scattering amplitude showed to be significantly different between the healthy part of the liver and the hepatic carcinoma (P < 0.05) being higher in normal liver tissue. A decrease in blood and lipid volume fractions and tissue oxygenation as well as an increase in scattering amplitude were observed when the tip of the needle crossed the margin from the healthy liver tissue to the carcinoma. The kurtosis for each derived clinical parameter was high in the hepatic tumor as compared with that in the healthy liver indicating intracarcinoma variability. CONCLUSIONS Tissue blood content, oxygenation level, lipid content, and tissue density all showed significant differences when the needle tip was guided from the healthy tissue to the carcinoma and can therefore be used to identify tissue boundaries during percutaneous image-guided interventions.

Relation between acid back -diffusion and luminal surface hydrophobicity in canine gastric mucosa: Effects of salicylate and prostaglandin

The stomach is thought to be protected from luminal acid by a gastric mucosal barrier that restricts the diffusion of acid into tissue. This study tested the hypothesis that the hydrophobic luminal surface of canine gastric mucosa incubated in Ussing chambers, impedes the back-diffusion of luminal acid into the tissue. Isolated sheets of mucosa were treated with cimetidine to inhibit spontaneous acid secretion, and incubated under conditions that prevented significant secretion of luminal bicarbonate. By measuring acid loss from the luminal compartment using the pH-stat technique, acid back-diffusion was continuously monitored; potential difference (PD) was measured as an index of tissue viability. Tissue luminal surface hydrophobicity was estimated by contact angle analysis at the end of each experiment. Addition of 16,16-dimethyl prostaglandin E$\sb2$ to the nutrient compartment enhanced luminal surface hydrophobicity, but did not reduce acid back-diffusion in tissues that maintained a constant PD. 10 mM salicylate at pH 4.00 in the luminal compartment reduced surface hydrophobicity, but this decrease did not occur if 1 ug/ml prostaglandin was present in the nutrient solution. Despite possessing relatively hydrophilic and relatively hydrophobic surface properties, respectively, acid back-diffusion in the absence of salicylate was not significantly different between these two groups. Neither group maintained a PD after incubation with salicylate. Lastly, radiolabelled salicylate was used to calculate the free (non-salicylate associated) acid loss in tissues incubated with salicylate and/or prostaglandin. No significant correlation was found between free acid back-diffusion and luminal surface hydrophobicity. These data do not support the hypothesis that acid back-diffusion in impeded by the hydrophobic surface presented by isolated canine gastric mucosa. ^

Great Barrier Reef coral element/Ca and stable isotope data and U-Th ages from IODP Expedition 325

Tropical south-western Pacific temperatures are of vital importance to the Great Barrier Reef (GBR), but the role of sea surface temperatures (SSTs) in the growth of the GBR since the Last Glacial Maximum remains largely unknown. Here we present records of Sr/Ca and d18O for Last Glacial Maximum and deglacial corals that show a considerably steeper meridional SST gradient than the present day in the central GBR. We find a 1-2 °C larger temperature decrease between 17° and 20°S about 20,000 to 13,000 years ago. The result is best explained by the northward expansion of cooler subtropical waters due to a weakening of the South Pacific gyre and East Australian Current. Our findings indicate that the GBR experienced substantial meridional temperature change during the last deglaciation, and serve to explain anomalous deglacial drying of northeastern Australia. Overall, the GBR developed through significant SST change and may be more resilient than previously thought.