Regulation and Release of Vasoactive Endoglin by Brain Endothelium in Response to Hypoxia/Reoxygenation in Stroke

In large vessel occlusion stroke, recanalization to restore cerebral perfusion is essential but not necessarily sufficient for a favorable outcome. Paradoxically, in some patients, reperfusion carries the risk of increased tissue damage and cerebral hemorrhage. Experimental and clinical data suggest that endothelial cells, representing the interface for detrimental platelet and leukocyte responses, likely play a crucial role in the phenomenon referred to as ischemia/reperfusion (I/R)-injury, but the mechanisms are unknown. We aimed to determine the role of endoglin in cerebral I/R-injury; endoglin is a membrane-bound protein abundantly expressed by endothelial cells that has previously been shown to be involved in the maintenance of vascular homeostasis. We investigated the expression of membranous endoglin (using Western blotting and RT-PCR) and the generation of soluble endoglin (using an enzyme-linked immunosorbent assay of cell culture supernatants) after hypoxia and subsequent reoxygenation in human non-immortalized brain endothelial cells. To validate these in vitro data, we additionally examined endoglin expression in an intraluminal monofilament model of permanent and transient middle cerebral artery occlusion in mice. Subsequently, the effects of recombinant human soluble endoglin were assessed by label-free impedance-based measurement of endothelial monolayer integrity (using the xCELLigence DP system) and immunocytochemistry. Endoglin expression is highly inducible by hypoxia in human brain endothelial monolayers in vitro, and subsequent reoxygenation induced its shedding. These findings were corroborated in mice during MCAO; an upregulation of endoglin was displayed in the infarcted hemispheres under occlusion, whereas endoglin expression was significantly diminished after transient MCAO, which is indicative of shedding. Of note is the finding that soluble endoglin induced an inflammatory phenotype in endothelial monolayers. The treatment of HBMEC with endoglin resulted in a decrease in transendothelial resistance and the downregulation of VE-cadherin. Our data establish a novel mechanism in which hypoxia triggers the initial endothelial upregulation of endoglin and subsequent reoxygenation triggers its release as a vasoactive mediator that, when rinsed into adjacent vascular beds after recanalization, can contribute to cerebral reperfusion injury.     

Metabolic effects in mice of cream formulation: Addition of both thickener and emulsifier does not alter lipid metabolism but modulates mucus cells and intestinal endoplasmic reticulum stress

Additives stabilize or improve the organoleptic or functional properties (or both) of many dairy products including whipping cream. Their influence on the metabolic effect of dairy cream is scarcely known. We tested the hypothesis that added emulsifier (lactic acid esters of mono- and diglycerides; MAG/DAG), thickener (carrageenan, CGN), or both, could modify the metabolic effect, notably in the intestine and liver. Nine-week-old male C57Bl/6J mice were fed UHT cream (indirect treatment) mixed with nonlipidic powder (final: 13% milkfat) for 1 or 4 wk. We compared creams (1) without additive (Ctl), (2) with thickener (Th), 0.02% of κ-CGN, and (3) with both thickener and emulsifier, 0.1% of MAG/DAG esters (Th/Em). We analyzed plasma parameters, intestine, and liver. Fasting glycemia, insulinemia, triglyceridemia, nonesterified fatty acids, body weight gain, and liver weight did not differ among groups. After 1 wk, Th/Em had higher expression in the duodenum of some of the genes involved in (1) intestinal lipid absorption and (2) tight junction proteins versus Ctl and Th. After 4 wk, mucus cell number in the small intestine was higher in Th/Em versus Ctl and Th. Genes involved in endoplasmic reticulum (ER) stress in the duodenum were more expressed in Th/Em after 1 wk. After 4 wk, in the colon, a higher expression of ER stress genes was observed for Th versus Th/Em and Ctl. Liver damage score was not altered by additives. Adding both CGN (0.02%) and MAG/DAG esters (0.1%) in dairy cream did not result in deleterious outcomes in mice after 4 wk regarding lipid metabolism, intestinal permeability, and liver disorders. The longer term effect of intestinal ER stress modulation deserves further investigation.     

Subclavian venogram as a guide to lead implantation

Melatonin synthesis in retinal photoreceptors is stimulated at night by a circadian oscillator and suppressed acutely by light. To identify photoreceptor mechanisms involved in the acute suppression of melatonin synthesis, an action spectrum was measured for dark-adapted Xenopus laevis eyecups at night. Intensity-response curves at six wavelengths from 400 to 650 nm were parallel, suggesting that a single photopigment predominates in melatonin suppression. Half-saturating intensities at 400, 440, 480, and 533 nm were not significantly different from one another, at 1-2 x 10(8) quanta cm(-2) s(-1). Significantly higher intensities of 580- and 650-nm light were required for melatonin suppression. These results indicate a predominant role for the principal green-absorbing rods in acute regulation of retinal melatonin synthesis in response to light, and argue against an important role for the red-absorbing cones. Higher than expected sensitivity at short wavelengths suggests that photoreceptors sensitive to blue and/or violet light may also contribute to melatonin suppression.     

Fitting in and feeling fine: Conformity and coping motives as mediators of the relationship between social anxiety and problematic drinking.

The present research was conducted to clarify the relationships among social anxiety, alcohol consumption, alcohol-related problems, and negative-reinforcement drinking motives among college students. Heavy drinking students (N = 316, 53.80% female) completed self-report measures of social anxiety, alcohol consumption, alcohol-related problems, and drinking motives. Findings indicated that students higher in social anxiety consumed less alcohol but experienced more negative consequences. Moreover, the relationship between social anxiety and negative consequences was mediated by coping and conformity drinking motives in addition to alcohol consumption. In the context of social anxiety, the current research demonstrates the importance of examining problematic drinking as distinct constructs: alcohol consumption and negative consequences. Findings are also discussed in terms of implications for interventions with socially anxious students. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Different metabolic and absorption patterns of betaine in response to dietary intake of whole-wheat grain, wheat aleurone or rye aleurone in catheterized pigs

In the present study, a novel method of measuring the uptake of nutrients by 1H NMR spectroscopy was performed to determine dietary effects. Six pigs catheterized in the portal vein and mesenteric artery were fed breads made from whole-wheat grain (WWG), wheat aleurone flour (WAF; 19?% aleurone of bread) or rye aleurone flour (RAF; 15?% aleurone of bread) in a repeated 3?×?3 crossover design. Three meals were provided daily (at 0, 5 and 10?h), and each period comprised of 1?week. Portal and arterial blood samples were collected at fasting (?30?min) on day 4–7, and on day 7, pooled blood samples were collected at 0–2.5, 2.5–5, 5–7.5 and 7.5–10?h after first (0?h) daily meal. Using the arterial-venous difference and ANOVA–simultaneous component analysis, plasma betaine was found to accumulate during the experimental period. On day 7, plasma betaine concentrations was 20–40?% higher than when feeding the WAF diet compared with RAF and WWG (P?=?0.001), whereas the content of betaine in the WAF diet was about twice as high. The arterial-venous difference showed a tendency (P?=?0.078) for the diet–time interaction of betaine absorption in the absorption phases but there was no diet effect (P?=?0.31), which indicates a steady-state absorption has been reached on day 7. Furthermore, high plasma betaine levels correlated with low plasma creatine of endogenous origin. In conclusion, the explorative method revealed that the plasma concentration and absorption of betaine were differentially modified by the diets.