A RAC-GEF network critical for early intestinal tumourigenesis.

RAC1 activity is critical for intestinal homeostasis, and is required for hyperproliferation driven by loss of the tumour suppressor gene Apc in the murine intestine. To avoid the impact of direct targeting upon homeostasis, we reasoned that indirect targeting of RAC1 via RAC-GEFs might be effective. Transcriptional profiling of Apc deficient intestinal tissue identified Vav3 and Tiam1 as key targets. Deletion of these indicated that while TIAM1 deficiency could suppress Apc-driven hyperproliferation, it had no impact upon tumourigenesis, while VAV3 deficiency had no effect. Intriguingly, deletion of either gene resulted in upregulation of Vav2, with subsequent targeting of all three (Vav2-/- Vav3-/- Tiam1-/-), profoundly suppressing hyperproliferation, tumourigenesis and RAC1 activity, without impacting normal homeostasis. Critically, the observed RAC-GEF dependency was negated by oncogenic KRAS mutation. Together, these data demonstrate that while targeting RAC-GEF molecules may have therapeutic impact at early stages, this benefit may be lost in late stage disease.

Contact line stick-slip motion and meniscus evolution on micrometer-size wavy fibres.

HYPOTHESIS: The architecture of complex-shaped fibres affects the motion of the contact line and the evolution of its associated menisci when a fibre is immersed into a liquid. Understanding and predicting the motion of the contact line is critical in the design of complex-shaped fibres for many engineering applications as well as for surface science. While wetting on classic circular cylinders has been well studied, singularities during the wetting process of complex-shaped fibres are not yet well understood. EXPERIMENTS: The dynamic wetting behaviour of axisymmetric sinus-shaped fibres immersed vertically in a liquid volume was investigated. Fibres were 3D-printed down to micrometre dimensions, and the Wilhelmy method was used in parallel with meniscus shape analysis. Moreover, a quasi-static theoretical model predicting the contact line movement and free energy of the system evolution on these fibres is also proposed. FINDINGS: The observation of liquid advancing and receding fronts highlighted a stick-slip motion of the meniscus depending on both the fibre surface curvature and its intrinsic wettability. The model predicts that the behaviour of the seemingly pinned and then jumping contact line, with associated changes in apparent contact angles, can be explained by the interplay between a constant local contact angle and the movement of the bulk liquid, leading to the storage of energy which is suddenly released when the contact line passes a given point of fibre curvature. Besides, acceleration/deceleration events that take place before and after the jumps are experimentally observed in good agreement with the model.

Multi-institutional dosimetric and geometric commissioning of image-guided small animal irradiators.

PURPOSE: To compare the dosimetric and geometric properties of a commercial x-ray based image-guided small animal irradiation system, installed at three institutions and to establish a complete and broadly accessible commissioning procedure. METHODS: The system consists of a 225 kVp x-ray tube with fixed field size collimators ranging from 1 to 44 mm equivalent diameter. The x-ray tube is mounted opposite a flat-panel imaging detector, on a C-arm gantry with 360° coplanar rotation. Each institution performed a full commissioning of their system, including half-value layer, absolute dosimetry, relative dosimetry (profiles, percent depth dose, and relative output factors), and characterization of the system geometry and mechanical flex of the x-ray tube and detector. Dosimetric measurements were made using Farmer-type ionization chambers, small volume air and liquid ionization chambers, and radiochromic film. The results between the three institutions were compared. RESULTS: At 225 kVp, with 0.3 mm Cu added filtration, the first half value layer ranged from 0.9 to 1.0 mm Cu. The dose-rate in-air for a 40 × 40 mm(2) field size, at a source-to-axis distance of 30 cm, ranged from 3.5 to 3.9 Gy/min between the three institutions. For field sizes between 2.5 mm diameter and 40 × 40 mm(2), the differences between percent depth dose curves up to depths of 3.5 cm were between 1% and 4% on average, with the maximum difference being 7%. The profiles agreed very well for fields >5 mm diameter. The relative output factors differed by up to 6% for fields larger than 10 mm diameter, but differed by up to 49% for fields ≤5 mm diameter. The mechanical characteristics of the system (source-to-axis and source-to-detector distances) were consistent between all three institutions. There were substantial differences in the flex of each system. CONCLUSIONS: With the exception of the half-value layer, and mechanical properties, there were significant differences between the dosimetric and geometric properties of the three systems. This underscores the need for careful commissioning of each individual system for use in radiobiological experiments.

Generalized inflammation during peritonitis evidenced by intracutaneous E-selectin expression.

E-selectin is a human endothelial leukocyte adhesion molecule which is expressed on endothelial cells after exposure to inflammatory mediators and which is known to be involved in the adhesion of polymorphonuclear cells to the endothelium in vitro. Data on E-selectin expression in vivo are limited. In the present report, we studied the expression of E-selectin in skin biopsies from patients with peritonitis due to a perforation of the gastrointestinal tract. Substantial E-selectin expression was observed on the vasculature of the skin from six out of eight of these severely ill patients. Skin obtained from healthy individuals stained negative, or showed a faint patchy staining in 30% of biopsies tested. These results provide evidence that E-selectin expression was induced at a distance from the primary inflammatory process on the vascular endothelium of the skin during severe peritonitis. Cutaneous E-selectin expression thus reflected on the surface of the body a state of generalized activated endothelium.