A Transgenic Model for Conditional Induction and Rescue of Portal Hypertension Reveals a Role of VEGF-Mediated Regulation of Sinusoidal Fenestrations

Portal hypertension (PH) is a common complication and a leading cause of death in patients with chronic liver diseases. PH is underlined by structural and functional derangement of liver sinusoid vessels and its fenestrated endothelium. Because in most clinical settings PH is accompanied by parenchymal injury, it has been difficult to determine the precise role of microvascular perturbations in causing PH. Reasoning that Vascular Endothelial Growth Factor (VEGF) is required to maintain functional integrity of the hepatic microcirculation, we developed a transgenic mouse system for a liver-specific-, reversible VEGF inhibition. The system is based on conditional induction and de-induction of a VEGF decoy receptor that sequesters VEGF and preclude signaling. VEGF blockade results in sinusoidal endothelial cells (SECs) fenestrations closure and in accumulation and transformation of the normally quiescent hepatic stellate cells, i.e. provoking the two processes underlying sinusoidal capillarization. Importantly, sinusoidal capillarization was sufficient to cause PH and its typical sequela, ascites, splenomegaly and venous collateralization without inflicting parenchymal damage or fibrosis. Remarkably, these dramatic phenotypes were fully reversed within few days from lifting-off VEGF blockade and resultant re-opening of SECs' fenestrations. This study not only uncovered an indispensible role for VEGF in maintaining structure and function of mature SECs, but also highlights the vasculo-centric nature of PH pathogenesis. Unprecedented ability to rescue PH and its secondary manifestations via manipulating a single vascular factor may also be harnessed for examining the potential utility of de-capillarization treatment modalities

The Use of Drones to Determine Rodent Location and Damage in Agricultural Crops

Rodent pests cause extensive damage to agricultural crops worldwide. Farmers’ ability to monitor rodent activity and damage within crops is limited due to their inability to simultaneously survey vast agricultural areas for rodent activity, the inability to enter certain fields, and the difficulty of monitoring rodent numbers, as well as using traps due to trap shyness and high labor costs. Drones can potentially be used to monitor rodent numbers and damage because they can cover large areas quickly without damaging crops and carry sensors that provide high-resolution imagery. Here, we investigated whether rodent activity (Levant voles Microtus guentheri and house mice Mus musculus) is related to vegetation health and biomass in Alfalfa (Medicago sativa) fields. We used a drone to photograph one hundred and twenty 10 × 10 m plots in nine fields and calculate the plots’ normalized difference vegetation index (NDVI) and biomass. On each plot, we also trapped rodents, counted rodent burrows, and evaluated the harvested dry crop yield. The number of burrows was positively related to the number of Levant voles trapped (F1,110 = 12.08, p 1,110 = 5.23, p 1,83 = 3.81, p 1,95 = 73.14, p 1,95 = 79.58, p < 0.001) were negatively related to the number of Levant voles trapped, and the number of burrows were not related to the number of house mice trapped. We demonstrate that drones can be used to assist farmers in determining the Levant vole presence and damage within crop fields to control rodents using precision agriculture methods, such as adding rodenticides in specific areas, thus increasing efficiency and decreasing the amount of pesticides used

The Use of Drones to Determine Rodent Location and Damage in Agricultural Crops

Rodent pests cause extensive damage to agricultural crops worldwide. Farmers&rsquo; ability to monitor rodent activity and damage within crops is limited due to their inability to simultaneously survey vast agricultural areas for rodent activity, the inability to enter certain fields, and the difficulty of monitoring rodent numbers, as well as using traps due to trap shyness and high labor costs. Drones can potentially be used to monitor rodent numbers and damage because they can cover large areas quickly without damaging crops and carry sensors that provide high-resolution imagery. Here, we investigated whether rodent activity (Levant voles Microtus guentheri and house mice Mus musculus) is related to vegetation health and biomass in Alfalfa (Medicago sativa) fields. We used a drone to photograph one hundred and twenty 10 &times; 10 m plots in nine fields and calculate the plots&rsquo; normalized difference vegetation index (NDVI) and biomass. On each plot, we also trapped rodents, counted rodent burrows, and evaluated the harvested dry crop yield. The number of burrows was positively related to the number of Levant voles trapped (F1,110 = 12.08, p &lt; 0.01) and negatively related to the number of house mice trapped (F1,110 = 5.23, p &lt; 0.05). Biomass extracted from drone images was positively related to the yield harvested by hand (F1,83 = 3.81, p &lt; 0.05). Farmers, therefore, can use burrow counting in place of trapping Levant voles, and biomass estimates from drones can be used in place of manual yield calculations. NDVI (F1,95 = 73.14, p &lt; 0.001) and biomass (F1,95 = 79.58, p &lt; 0.001) were negatively related to the number of Levant voles trapped, and the number of burrows were not related to the number of house mice trapped. We demonstrate that drones can be used to assist farmers in determining the Levant vole presence and damage within crop fields to control rodents using precision agriculture methods, such as adding rodenticides in specific areas, thus increasing efficiency and decreasing the amount of pesticides used

VEGF modulates early heart valve formation

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Conditional switching of VEGF provides new insights into adult neovascularization and pro-angiogenic therapy

To gain insight into neovascularization of adult organs and to uncover inherent obstacles in vascular endothelial growth factor (VEGF)-based therapeutic angiogenesis, a transgenic system for conditional switching of VEGF expression was devised. The system allows for a reversible induction of VEGF specifically in the heart muscle or liver at any selected schedule, thereby circumventing embryonic lethality due to developmental misexpression of VEGF. Using this system, we demonstrate a progressive, unlimited ramification of the existing vasculature. In the absence of spatial cues, however, abnormal vascular trees were produced, a consequence of chaotic connections with the existing network and formation of irregularly shaped sac-like vessels. VEGF also caused a massive and highly disruptive edema. Importantly, premature cessation of the VEGF stimulus led to regression of most acquired vessels, thus challenging the utility of therapeutic approaches relying on short stimulus duration. A critical transition point was defined beyond which remodeled new vessels persisted for months after withdrawing VEGF, conferring a long-term improvement in organ perfusion. This novel genetic system thus highlights remaining problems in the implementation of pro-angiogenic therapy

Conditional islet hypovascularisation does not preclude beta cell expansion during pregnancy in mice

Endothelial-endocrine cell interactions and vascular endothelial growth factor (VEGF)-A signalling are deemed essential for maternal islet vascularisation, glucose control and beta cell expansion during mouse pregnancy. The aim of this study was to assess whether pregnancy-associated beta cell expansion was affected under conditions of islet hypovascularisation. Soluble fms-like tyrosine kinase 1 (sFLT1), a VEGF-A decoy receptor, was conditionally overexpressed in maternal mouse beta cells from 1.5 to 14.5 days post coitum. Islet vascularisation, glycaemic control, beta cell proliferation, individual beta cell size and total beta cell volume were assessed in both pregnant mice and non-pregnant littermates. Conditional overexpression of sFLT1 in beta cells resulted in islet hypovascularisation and glucose intolerance in both pregnant and non-pregnant mice. In contrast to non-pregnant littermates, glucose intolerance in pregnant mice was transient. sFLT1 overexpression did not affect pregnancy-associated changes in beta cell proliferation, individual beta cell size or total beta cell volume. Reduced intra-islet VEGF-A signalling results in maternal islet hypovascularisation and impaired glycaemic control but does not preclude beta cell expansion during mouse pregnancy

Role of HIF-1alpha in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis.

As a result of deprivation of oxygen (hypoxia) and nutrients, the growth and viability of cells is reduced. Hypoxia-inducible factor (HIF)-1alpha helps to restore oxygen homeostasis by inducing glycolysis, erythropoiesis and angiogenesis. Here we show that hypoxia and hypoglycaemia reduce proliferation and increase apoptosis in wild-type (HIF-1alpha+/+) embryonic stem (ES) cells, but not in ES cells with inactivated HIF-1alpha genes (HIF-1alpha-/-); however, a deficiency of HIF-1alpha does not affect apoptosis induced by cytokines. We find that hypoxia/hypoglycaemia-regulated genes involved in controlling the cell cycle are either HIF-1alpha-dependent (those encoding the proteins p53, p21, Bcl-2) or HIF-1alpha-independent (p27, GADD153), suggesting that there are at least two different adaptive responses to being deprived of oxygen and nutrients. Loss of HIF-1alpha reduces hypoxia-induced expression of vascular endothelial growth factor, prevents formation of large vessels in ES-derived tumours, and impairs vascular function, resulting in hypoxic microenvironments within the tumour mass. However, growth of HIF-1alpha tumours was not retarded but was accelerated, owing to decreased hypoxia-induced apoptosis and increased stress-induced proliferation. As hypoxic stress contributes to many (patho)biological disorders, this new role for HIF-1alpha in hypoxic control of cell growth and death may be of general pathophysiological importance

Loss of HIF-2alpha and inhibition of VEGF impair fetal lung maturation, whereas treatment with VEGF prevents fatal respiratory distress in premature mice

Respiratory distress syndrome (RDS) due to insufficient production of surfactant is a common and severe complication of preterm delivery. Here, we report that loss of the hypoxia-inducible transcription factor-2alpha (HIF-2alpha) caused fatal RDS in neonatal mice due to insufficient surfactant production by alveolar type 2 cells. VEGF, a target of HIF-2alpha, regulates fetal lung maturation: because VEGF levels in alveolar cells were reduced in HIF-2alpha-deficient fetuses; mice with a deficiency of the VEGF(164) and VEGF(188) isoforms or of the HIF-binding site in the VEGF promotor died of RDS; intrauterine delivery of anti-VEGF-receptor-2 antibodies caused RDS and VEGF stimulated production of surfactant proteins by cultured type 2 pneumocytes. Intrauterine delivery or postnatal intratracheal instillation of VEGF stimulated conversion of glycogen to surfactant and protected preterm mice against RDS. The pneumotrophic effect of VEGF may have therapeutic potential for lung maturation in preterm infants.status: publishe