Endothelial Mechanosignaling: Does One Sensor Fit All?

Significance: Forces are important in the cardiovascular system, acting as regulators of vascular physiology and pathology. Residing at the blood vessel interface, cells (endothelial cell, EC) are constantly exposed to vascular forces, including shear stress. Shear stress is the frictional force exerted by blood flow, and its patterns differ based on vessel geometry and type. These patterns range from uniform laminar flow to nonuniform disturbed flow. Although ECs sense and differentially respond to flow patterns unique to their microenvironment, the mechanisms underlying endothelial mechanosensing remain incompletely understood. Recent Advances: A large body of work suggests that ECs possess many mechanosensors that decorate their apical, junctional, and basal surfaces. These potential mechanosensors sense blood flow, translating physical force into biochemical signaling events. Critical Issues: Understanding the mechanisms by which proposed mechanosensors sense and respond to shear stress requires an integrative approach. It is also critical to understand the role of these mechanosensors not only during embryonic development but also in the different vascular beds in the adult. Possible cross talk and integration of mechanosensing via the various mechanosensors remain a challenge. Future Directions: Determination of the hierarchy of endothelial mechanosensors is critical for future work, as is determination of the extent to which mechanosensors work together to achieve force-dependent signaling. The role and primary sensors of shear stress during development also remain an open question. Finally, integrative approaches must be used to determine absolute mechanosensory function of potential mechanosensors. Antioxid. Redox Signal. 25, 373–388

Hemodynamic Mechanisms Regulating Inflammatory Vascular Remodeling

Hemodynamic forces are critical regulators of vascular health and disease. Shear stress, the frictional force of blood flowing over the endothelium, is a major hemodynamic input into vascular function. For example, inflammatory blood flow patterns induce programs of intercellular signaling and gene expression that cause local vascular inflammation. Endothelial cells mediate the interactions between blood flow and blood vessels, and as such are exquisitely sensitive to forces. Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1), a well-known endothelial mechanosensor, is required for flow-induced endothelial inflammation, yet the signaling pathways connecting hemodynamics, PECAM-1, and vascular inflammation remain to be elucidated. This dissertation provides mechanistic insight into two PECAM-1 dependent inflammatory pathways: assembly of fibronectin (FN) into fibrils and the role of the adaptor protein Shc in inflammatory vascular remodeling. FN is an extracellular matrix protein that plays major roles in vascular development and disease. Required for embryonic development of the vascular plexus and heart, FN is also heavily deposited in atherosclerotic plaques and chronically inflamed vessels. As FN is known to sustain endothelial inflammation, knowledge of its assembly is critical to understanding vascular disease. In Chapter 2, I demonstrate that FN assembly is controlled by altered hemodynamics via PECAM-1. Additionally, I delineate a mechanism by which the small GTPase RhoA and β1 integrins regulate FN assembly in a PECAM-1 dependent manner. This is the first time that FN assembly has been mechanistically linked to hemodynamics, and suggests a route by which chronic inflammation may take hold in the endothelium. Vascular inflammation may result in remodeling of blood vessels, and depending on the physical cues at work, this remodeling may result in widening or narrowing of the blood vessels. The adaptor protein Shc is a known regulator of endothelial inflammation and outward vascular remodeling, and is phosphorylated upon the onset of shear. In Chapter 3, I demonstrate that Shc tyrosine phosphorylation is required for outward collateral remodeling in response to femoral artery ligation. I also demonstrate that Shc is required for atherogenesis, which is pathological inward remodeling of arteries in response to inflammatory flow. Critically, I show the involvement of Shc in atherogenesis to be responsive to changes in hemodynamics. These data suggest that Shc is a major regulator of vascular remodeling, as well as suggest potential targets for downstream vascular therapeutics.Doctor of Philosoph

Haemodynamics Regulate Fibronectin Assembly via PECAM

Fibronectin (FN) assembly and fibrillogenesis are critically important in both development and the adult organism, but their importance in vascular functions is not fully understood. Here we identify a novel pathway by which haemodynamic forces regulate FN assembly and fibrillogenesis during vascular remodelling. Induction of disturbed shear stress in vivo and in vitro resulted in complex FN fibril assembly that was dependent on the mechanosensor PECAM. Loss of PECAM also inhibited the cell-intrinsic ability to remodel FN. Gain- and loss-of-function experiments revealed that PECAM-dependent RhoA activation is required for FN assembly. Furthermore, PECAM−/− mice exhibited reduced levels of active β1 integrin that were responsible for reduced RhoA activation and downstream FN assembly. These data identify a new pathway by which endothelial mechanotransduction regulates FN assembly and flow-mediated vascular remodelling

Effect of milk type and processing on iodine concentration of organic and conventional winter milk at retail: implications for nutrition

Milk is the largest source of iodine in UK diets and an earlier study showed that organic summer milk had significantly lower iodine concentration than conventional milk. There are no comparable studies with winter milk or the effect of milk fat class or heat processing method. Two retail studies with winter milk are reported. Study 1 showed no effect of fat class but organic milk was 32.2% lower in iodine than conventional milk (404 vs. 595 μg/L; P < 0.001). Study 2 found no difference between conventional and Channel Island milk but organic milk contained 35.5% less iodine than conventional milk (474 vs. 306 μg/L; P < 0.001). UHT and branded organic milk also had lower iodine concentrations than conventional milk (331 μg/L; P < 0.001 and 268 μg/L: P < 0.0001 respectively). The results indicate that replacement of conventional milk by organic or UHT milk will increase the risk of sub-optimal iodine status especially for pregnant/lactating women

Differential effects of oilseed supplements on methane production and milk fatty acid concentrations in dairy cows

It is known that supplementing dairy cow diets with full-fat oilseeds can be used as a strategy to mitigate methane emissions, through their action on rumen fermentation. However, direct comparisons of the effect of different oil sources are very few, as are studies implementing supplementation levels that reflect what is commonly fed on commercial farms. The objective was to investigate the effect of feeding different forms of supplemental plant oils on both methane emissions and milk fatty acid (FA) profile. Four multiparous, Holstein-Friesian cows in mid-lactation were randomly allocated to one of four treatment diets in a 4 x 4 Latin square design with 28-day periods. Diets were fed as a TMR with a 50:50 forage:concentrate ratio (dry matter, DM basis) with the forage consisting of 75:25 maize silage:grass silage (DM). Dietary treatments were a control diet containing no supplemental fat, and three treatment diets containing extruded linseed (EL), calcium salts of palm and linseed oil (CPLO) or milled rapeseed (MR) formulated to provide each cow with an estimated 500 g additional oil/d (22 g oil/kg diet DM). Dry matter intake (DMI), milk yield, milk composition and methane production were measured at the end of each experimental period when cows were housed in respiration chambers for 4 days. There was no effect of treatment diet on DMI or milk protein or lactose concentration, but oilseed-based supplements increased milk yield compared with the control diet and milk fat concentration relative to control was reduced by 4 g/kg by supplemental EL. Feeding CPLO reduced methane production, and both linseed-based supplements decreased methane yield (by 1.8 L/kg DMI) and intensity (by 2.7 L/kg milk yield) compared with the control diet, but feeding MR had no effect on methane emission. All the fat supplements decreased milk total saturated fatty acid (SFA) concentration compared with the control, and SFA were replaced with mainly cis-9 18:1 but also trans FA (and in the case of EL and CPLO there were increases in polyunsaturated FA concentration). Supplementing dairy cow diets with these oilseed-based preparations affected milk FA profile and increased milk yield. However, only the linseed-based supplements reduced methane production, yield, or intensity, whilst feeding MR had no effect

Cardiac contraction activates endocardial Notch signaling to modulate chamber maturation in zebrafish

Congenital heart disease often features structural abnormalities that emerge during development. Accumulating evidence indicates a crucial role for cardiac contraction and the resulting fluid forces in shaping the heart, yet the molecular basis of this function is largely unknown. Using the zebrafish as a model of early heart development, we investigated the role of cardiac contraction in chamber maturation, focusing on the formation of muscular protrusions called trabeculae. By genetic and pharmacological ablation of cardiac contraction, we showed that cardiac contraction is required for trabeculation through its role in regulating notch1b transcription in the ventricular endocardium. We also showed that Notch1 activation induces expression of ephrin b2a (efnb2a) and neuregulin 1 (nrg1) in the endocardium to promote trabeculation and that forced Notch activation in the absence of cardiac contraction rescues efnb2a and nrg1 expression. Using in vitro and in vivo systems, we showed that primary cilia are important mediators of fluid flow to stimulate Notch expression. Together, our findings describe an essential role for cardiac contraction-responsive transcriptional changes in endocardial cells to regulate cardiac chamber maturation

Further studies on the iodine concentration of conventional, organic and UHT semi-skimmed milk at retail in the UK

Milk is the largest source of iodine in UK diets and earlier studies showed organic summer and winter milk to be significantly lower in iodine than conventional milk. One study also showed UHT milk to have lower iodine concentration. The study on winter and UHT milk was small and accordingly a new study is reported here involving conventional, organic and UHT semi-skimmed milk from four supermarkets over a six-month period in summer and winter in two regions of the UK. The results showed organic milk to be 44% lower in iodine than conventional milk (427 vs. 241 mg/L, P < 0.001) and UHT milk was 27% lower in iodine than conventional milk (427 vs. 314 mg/L, P < 0.001) although the differences tended to be less in the summer. The results indicate that replacement of conventional milk by organic or UHT milk will increase the risk of sub-optimal iodine status especially for pregnant/lactating women

Further studies on the iodine concentration of conventional, organic and UHT semi-skimmed milk at retail in the UK

Milk is the largest source of iodine in UK diets and earlier studies showed organic summer and winter milk to be significantly lower in iodine than conventional milk. One study also showed UHT milk to have lower iodine concentration. The study on winter and UHT milk was small and accordingly a new study is reported here involving conventional, organic and UHT semi-skimmed milk from four supermarkets over a six-month period in summer and winter in two regions of the UK. The results showed organic milk to be 44% lower in iodine than conventional milk (427 vs. 241 mg/L, P < 0.001) and UHT milk was 27% lower in iodine than conventional milk (427 vs. 314 mg/L, P < 0.001) although the differences tended to be less in the summer. The results indicate that replacement of conventional milk by organic or UHT milk will increase the risk of sub-optimal iodine status especially for pregnant/lactating women