The effect of matrix metalloproteinase 2 and matrix metalloproteinase 2/9 deletion in experimental post-thrombotic vein wall remodeling.

BACKGROUND: Vein wall fibrotic injury following deep venous thrombosis (VT) is associated with elevated matrix metalloproteinases (MMPs). Whether and by what mechanism MMP2 contributes to vein wall remodeling after VT is unknown. METHODS: Stasis VT was produced by ligation of the inferior vena cava and tissue was harvested at 2, 8, and 21 days in MMP2 -/- and genetic wild type (WT) mice. Tissue analysis by immunohistochemistry, enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and zymography was performed. RESULTS: Thrombus resolution was less at 8 days in MMP2 -/- compared with WT, evidenced by a 51% increase in VT size (P < .01), and threefold fewer von Willebrand's factor positive channels (P < .05). In MMP2 -/- mice, the main phenotypic fibrotic differences occurred at 8 days post-VT, with significantly less vein wall collagen content (P = .013), fourfold lower procollagen III gene expression (P < .01), but no difference in procollagen I compared with WT. Decreased inflammation in MMP2 -/- vein walls was suggested by ∼ threefold reduced TNFα and IL-1ß at 2 days and 8 days post-VT (P < .05). A fourfold increase in vein wall monocytes (P = .03) with threefold decreased apoptosis (P < .05), but no difference in cellular proliferation at 8 days was found in MMP2 -/- compared with WT. As increased compensatory MMP9 activity was observed in the MMP2 -/-mice, MMP2/9 double null mice had thrombus induced with VT harvest at 8 days. Consistently, twofold larger VT, a threefold decrease in vein wall collagen, and a threefold increase in monocytes were found (all P < .05). Similar findings were observed in MMP9 -/- mice administered an exogenous MMP2 inhibitor. CONCLUSIONS: In stasis VT, deletion of MMP2 was associated with less midterm vein wall fibrosis and inflammation, despite an increase in monocytes. Consideration that VT resolution was impaired with MMP2 (and MMP2/9) deletion suggests direct inhibition will likely also require anticoagulant therapy.

Inclusion of Food Safety Information in Home-delivered U.K. Meal-kit Recipes.

Home-delivered meal-kit recipe boxes provide consumers with fresh, measured ingredients requiring storage, preparation, and cooking by following a recipe card.Previous literature has suggested that including food safety information within recipes may positively impact behavior. Therefore, there is a need to better understand what food safety information is included on the meal-kit recipe cards. Content analysis was performed on U.K. meal-kit provider recipe cards (n = 359) obtained using citizen science methods. Although 46% of recipes referred to handwashing at the start of recipe preparation, these stated 'wash hands' with no further advice regarding hand hygiene, and half (48%) did not refer to handwashing during recipe preparation. Most recipes included produce (99%) and referred to washing fruit and vegetables (88%). For meal-kits that provided animal proteins (n = 332), 50% referred to storing ingredients in the fridge, and only one recipe referred to recommended temperatures (≤5°C). Where applicable (n = 346), food safety advice to prevent cross-contamination was present in 51% of recipes.Statements concerning cooking adequacy of animal proteins (n = 1306) included subjective cooking guidance, with 35% referring to visual assessment of color and 26% cooking duration. For best practice food safety guidance, two recipes referred to end-point temperature, and one stated to use a temperature probe. While all meal-kit providers provided some food safety-related information in reviewed recipes, information was often not sufficient to inform consumers about food safety risk-reducing behaviors in the domestic setting. Observational research is needed to understand consumer engagement with how food safety information in meal-kit recipes impacts the behavior of consumers.

Toll-like receptor 9 signaling is critical for early experimental deep vein thrombosis resolution.

OBJECTIVE: Toll-like receptors (TLR) bridge innate immunity and host responses, including inflammation. Sterile inflammation such as a venous thrombus (Vt) may involve TLR signaling, including TLR9. METHODS AND RESULTS: TLR9 signaling on thrombus resolution was investigated using a mouse model of stasis Vt. Vt were significantly larger in TLR9-/- mice compared with wild-type (WT) at 2 and 8 days, despite a 2-fold increase in thrombus polymorphonucleic neutrophils at 2 days and monocytes at 8 days, whereas thrombus collagen and neovascularization was 55% and 37% less, respectively, at 8 days. Coincidently, decreased fibrinogen and increased thrombin-antithrombin complex were observed in TLR9-/- mouse thrombi. Vein wall interferon-α, interleukin-1α, and interleukin-2 were significantly reduced in TLR9-/- mice compared with WT. Thrombus cell death pathway markers were not significantly altered at 2 days, but caspase-1 was reduced in TLR9-/- thrombi at 8 days. MyD88 confers TLR9 intracellular signaling, but MyD88-/- mice had Vt resolution similar to that of WT. However, inhibition of the NOTCH ligand δ-like 4 was associated with larger Vt. Finally, stimulation with a TLR9 agonist was associated with smaller Vt. CONCLUSIONS: TLR9 signaling is integral for early and mid-Vt resolution through modulation of sterile inflammation, maintaining a TH1 milieu, and effects on the thrombosis pathway.

Ionizing radiation-induced bystander mutagenesis and adaptation: quantitative and temporal aspects.

This work explores several quantitative aspects of radiation-induced bystander mutagenesis in WTK1 human lymphoblast cells. Gamma-irradiation of cells was used to generate conditioned medium containing bystander signals, and that medium was transferred onto naïve recipient cells. Kinetic studies revealed that it required up to 1h to generate sufficient signal to induce the maximal level of mutations at the thymidine kinase locus in the bystander cells receiving the conditioned medium. Furthermore, it required at least 1h of exposure to the signal in the bystander cells to induce mutations. Bystander signal was fairly stable in the medium, requiring 12-24h to diminish. Medium that contained bystander signal was rendered ineffective by a 4-fold dilution; in contrast a greater than 20-fold decrease in the cell number irradiated to generate a bystander signal was needed to eliminate bystander-induced mutagenesis. This suggested some sort of feedback inhibition by bystander signal that prevented the signaling cells from releasing more signal. Finally, an ionizing radiation-induced adaptive response was shown to be effective in reducing bystander mutagenesis; in addition, low levels of exposure to bystander signal in the transferred medium induced adaptation that was effective in reducing mutations induced by subsequent gamma-ray exposures.

The Perceived Size and Shape of Objects in Peripheral Vision.

Little is known about how we perceive the size and shape of objects in far peripheral vision. Observations made during an artistic study of visual space suggest that objects appear smaller and compressed in the periphery compared with central vision. To test this, we conducted three experiments. In Experiment 1, we asked participants to draw how a set of peripheral discs appeared when viewed peripherally without time or eye movement constraints. In Experiment 2, we used the method of constant stimuli to measure when a briefly presented peripheral stimulus appeared bigger or smaller compared with a central fixated one. In Experiment 3, we measured how accurate participants were in discriminating shapes presented briefly in the periphery. In Experiment 1, the peripheral discs were reported as appearing significantly smaller than the central disc, and as having an elliptical or polygonal contour. In Experiment 2, participants judged the size of peripheral discs as being significantly smaller when compared with the central disc across most of the peripheral field, and in Experiment 3, participants were quite accurate in reporting the shape of the peripheral object, except in the far periphery. Our results show that objects in the visual periphery are perceived as diminished in size when presented for long and brief exposures, suggesting diminution is an intrinsic feature of the structure of the visual space. Shape distortions, however, are reported only with longer exposures.

Comparing artistic and geometrical perspective depictions of space in the visual field.

Which is the most accurate way to depict space in our visual field? Linear perspective, a form of geometrical perspective, has traditionally been regarded as the correct method of depicting visual space. But artists have often found it is limited in the angle of view it can depict; wide-angle scenes require uncomfortably close picture viewing distances or impractical degrees of enlargement to be seen properly. Other forms of geometrical perspective, such as fisheye projections, can represent wider views but typically produce pictures in which objects appear distorted. In this study we created an artistic rendering of a hemispherical visual space that encompassed the full visual field. We compared it to a number of geometrical perspective projections of the same space by asking participants to rate which best matched their visual experience. We found the artistic rendering performed significantly better than the geometrically generated projections.

Urokinase plasminogen activator independent early experimental thrombus resolution: MMP2 as an alternative mechanism.

Deep-vein thrombosis (DVT) resolution is thought to be primarily a urokinase plasminogen activator (uPA) -dependent mechanism, although observations suggest other non-fibrinolytic mechanisms may exist. We explored the role of matrix metalloproteinase (MMP) -2 and -9 in early DVT resolution in uPA-deficient mice. Male B6/SVEV (WT) and genetically matched uPA -/- mice underwent inferior vena cava (IVC) ligation to create stasis venous thrombi, with IVC and thrombus harvest. Thrombus size was similar between WT and uPA -/- mice at day 4, suggesting early non uPA-dependent resolution. Intrathrombus neutrophils and monocytes were reduced 3- and 3.5-fold in uPA -/- mice as compared with WT. By ELISA, tumour necrosis factor α and interleukin 1ß were not altered, while interferon (IFN)γ was significantly elevated in uPA -/- mice. A compensatory increase in thrombus tPA was not observed, plasmin activity was reduced and PAI-1 was elevated 2.5-fold in uPA -/- mice. Active MMP2, but not MMP9, was elevated 3-fold in uPA -/- mice as compared with WT as well as MMP-14, an MMP2 activator. Collagen type IV and fibrinogen were reduced in uPA -/- mice thrombi as compared with WT. IFNγ induces MMP2, and blockade of IFNγ was associated with larger venous thrombi and reduced active MMP2, as compared with WT. Consistently, MMP2 -/- mice had larger VT as compared with WT controls, despite normal thrombus plasmin levels. Taken together, early experimental venous thrombus resolution is independent of uPA, and, in part, inflammatory cell influx. MMP2-dependent thrombolysis is an important compensatory mechanism of venous thrombus resolution, possibly by collagen type IV metabolism, and may represent an exploitable therapeutic avenue.