Targeting lymphatic vessel functions through tyrosine kinases

The lymphatic vascular system is actively involved in tissue fluid homeostasis, immune surveillance and fatty acid transport. Pathological conditions can arise from injury to the lymphatics, or they can be recruited in the context of cancer to facilitate metastasis. Protein tyrosine kinases are central players in signal transduction networks and regulation of cell behavior. In the lymphatic endothelium, tyrosine kinases are involved in processes such as the maintenance of existing lymphatic vessels, growth and maturation of new vessels and modulation of their identity and function. As such, they are attractive targets for both existing inhibitors and the development of new inhibitors which affect lymphangiogenesis in pathological states such as cancer. RNAi screening provides an opportunity to identify the functional role of tyrosine kinases in the lymphatics. This review will discuss the role of tyrosine kinases in lymphatic biology and the potential use of inhibitors for anti-lymphangiogenic therapy

Differential receptor binding and regulatory mechanisms for the lymphangiogenic growth factors VEGF-C and VEGF-D

VEGF-C and VEGF-D are secreted glycoproteins that induce angiogenesis and lymphangiogenesis in cancer, thereby promoting tumor growth and spread. They exhibit structural homology and activate VEGFR-2 and VEGFR-3, receptors on endothelial cells that signal for growth of blood vessels and lymphatics. VEGF-C and VEGF-D were thought to exhibit similar bioactivities, yet recent studies indicated distinct signaling mechanisms (e.g. tumor-derived VEGF-C promoted expression of the prostaglandin biosynthetic enzyme COX-2 in lymphatics, a response thought to facilitate metastasis via the lymphatic vasculature, whereas VEGF-D did not). Here we explore the basis of the distinct bioactivities of VEGF-D using a neutralizing antibody, peptide mapping, and mutagenesis to demonstrate that the N-terminal α-helix of mature VEGF-D (Phe(93)–Arg(108)) is critical for binding VEGFR-2 and VEGFR-3. Importantly, the N-terminal part of this α-helix, from Phe(93) to Thr(98), is required for binding VEGFR-3 but not VEGFR-2. Surprisingly, the corresponding part of the α-helix in mature VEGF-C did not influence binding to either VEGFR-2 or VEGFR-3, indicating distinct determinants of receptor binding by these growth factors. A variant of mature VEGF-D harboring a mutation in the N-terminal α-helix, D103A, exhibited enhanced potency for activating VEGFR-3, was able to promote increased COX-2 mRNA levels in lymphatic endothelial cells, and had enhanced capacity to induce lymphatic sprouting in vivo. This mutant may be useful for developing protein-based therapeutics to drive lymphangiogenesis in clinical settings, such as lymphedema. Our studies shed light on the VEGF-D structure/function relationship and provide a basis for understanding functional differences compared with VEGF-C

Solidification of Al-Sn-Cu based immiscible alloys under intense shearing

The official published version of the Article can be accessed from the link below - Copyright @ 2009 The Minerals, Metals & Materials Society and ASM InternationalThe growing importance of Al-Sn based alloys as materials for engineering applications necessitates the development of uniform microstructures with improved performance. Guided by the recently thermodynamically assessed Al-Sn-Cu system, two model immiscible alloys, Al-45Sn-10Cu and Al-20Sn-10Cu, were selected to investigate the effects of intensive melt shearing provided by the novel melt conditioning by advanced shear technology (MCAST) unit on the uniform dispersion of the soft Sn phase in a hard Al matrix. Our experimental results have confirmed that intensive melt shearing is an effective way to achieve fine and uniform dispersion of the soft phase without macro-demixing, and that such dispersed microstructure can be further refined in alloys with precipitation of the primary Al phase prior to the demixing reaction. In addition, it was found that melt shearing at 200 rpm and 60 seconds will be adequate to produce fine and uniform dispersion of the Sn phase, and that higher shearing speed and prolonged shearing time can only achieve minor further refinement.This work is funded by the EPSRC and DT

Development Of Al-B-C Master Alloy Under External Fields

This study investigates the application of external fields in the development of an Al-B-C alloy, with the aim of synthesizing in situ Al3BC particles. A combination of ultrasonic cavitation and distributive mixing was applied for uniform dispersion of insoluble graphite particles in the Al melt, improving their wettability and its subsequent incorporation into the Al matrix. Lower operating temperatures facilitated the reduction in the amount of large clusters of reaction phases, with Al3BC being identified as the main phase in XRD analysis. The distribution of Al3BC particles was quantitatively evaluated. Grain refinement experiments reveal that Al-B-C alloy can act as a master alloy for Al-4Cu and AZ91D alloys, with average grain size reduction around 50% each at 1wt%Al-1.5B-2C additions

Solidification behavior of intensively sheared hypoeutectic Al-Si alloy liquid

The official published version of this article can be found at the link below.The effect of the processing temperature on the microstructural and mechanical properties of Al-Si (hypoeutectic) alloy solidified from intensively sheared liquid metal has been investigated systematically. Intensive shearing gives a significant refinement in grain size and intermetallic particle size. It also is observed that the morphology of intermetallics, defect bands, and microscopic defects in high-pressure die cast components are affected by intensive shearing the liquid metal. We attempt to discuss the possible mechanism for these effects.Funded by the EPSRC

The P72R Polymorphism in R248Q/W p53 Mutants Modifies the Mutant Effect on Epithelial to Mesenchymal Transition Phenotype and Cell Invasion via CXCL1 Expression

High-grade serous carcinoma (HGSC), the most lethal subtype of epithelial ovarian cancer (EOC), is characterized by widespread TP53 mutations (\u3e90%), most of which are missense mutations (\u3e70%). The objective of this study was to investigate differential transcriptional targets affected by a common germline P72R SNP (rs1042522) in two p53 hotspot mutants, R248Q and R248W, and identify the mechanism through which the P72R SNP affects the neomorphic properties of these mutants. Using isogenic cell line models, transcriptomic analysis, xenografts, and patient data, we found that the P72R SNP modifies the effect of p53 hotspot mutants on cellular morphology and invasion properties. Most importantly, RNA sequencing studies identified CXCL1 a critical factor that is differentially affected by P72R SNP in R248Q and R248W mutants and is responsible for differences in cellular morphology and functional properties observed in these p53 mutants. We show that the mutants with the P72 SNP promote a reversion of the EMT phenotype to epithelial characteristics, whereas its R72 counterpart promotes a mesenchymal transition via the chemokine CXCL1. These studies reveal a new role of the P72R SNP in modulating the neomorphic properties of p53 mutants via CXCL1, which has significant implications for tumor invasion and metastasis

Processing of ultrafine-size particulate metal matrix composites by advanced shear technology

Copyright @ 2009 ASM International. This paper was published in Metallurgical & Materials Transactions A 40A(3) and is made available as an electronic reprint with the permission of ASM International. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplications of any material in this paper for a fee or for commercial purposes, or modification of the content of this paper are prohibited.Lack of efficient mixing technology to achieve a uniform distribution of fine-size reinforcement within the matrix and the high cost of producing components have hindered the widespread adaptation of particulate metal matrix composites (PMMCs) for engineering applications. A new rheo-processing method, the melt-conditioning high-pressure die-cast (MC-HPDC) process, has been developed for manufacturing near-net-shape components of high integrity. The MC-HPDC process adapts the well-established high shear dispersive mixing action of a twin-screw mechanism to the task of overcoming the cohesive force of the agglomerates under a high shear rate and high intensity of turbulence. This is followed by direct shaping of the slurry into near-net-shape components using an existing cold-chamber die-casting process. The results indicate that the MC-HPDC samples have a uniform distribution of ultrafine-sized SiC particles throughout the entire sample in the as-cast condition. Compared to those produced by conventional high-pressure die casting (HPDC), MC-HPDC samples have a much improved tensile strength and ductility.EP-SR

Clinical study of the factors affecting radioulnar deviation of the wrist joint

<p>Abstract</p> <p>Background</p> <p>The radioulnar carpal joint is critical for hand and wrist function. Radioulnar deviation indicates distal radioulnar joint flexibility and reflects the structure and function of the carpal bones, ulna, radius and ligaments. The present study examined whether radioulnar deviation is affected by gender, manual labor, playing a musical instrument, playing sport, handedness, previous fracture or prior inflammation. The study used clinical findings based on anatomical landmarks</p> <p>Methods</p> <p>The ulnar, radial and total deviations for both left and right hands were measured in 300 subjects (157 men and 143 women) of mean age 21.7 years. Measurements were made with the forearm in a fixed pronated position using a novel specially designed goniometer. The gender of each subject was recorded, and information on playing of sport, playing a musical instrument, manual labor, handedness, and history of fracture or inflammation was sought. Data were analyzed using a multifactor ANOVA test.</p> <p>Results</p> <p>No statistically significant difference (p-value > 0.05) was found between those comparing groups except the total deviation of athletes' left hand versus the total deviation of non athletes' left hand (p-value 0.041 < 0.05) and the radial deviation of manual workers' left hand and non manual workers' left hand (p-value 0.002 < 0.05).</p> <p>Conclusions</p> <p>This study was based on clinical findings using anatomical landmarks. We found that manual workers and athletes showed greater left hand flexibility. This suggests that activities that place chronic stress on the radiocarpal joint can independently affect radioulnar deviation.</p

The Interplay Between Lymphatic Vessels and Chemokines

Chemokines are a family of small protein cytokines that act as chemoattractants to migrating cells, in particular those of the immune system. They are categorized functionally as either homeostatic, constitutively produced by tissues for basal levels of cell migration, or inflammatory, where they are generated in association with a pathological inflammatory response. While the extravasation of leukocytes via blood vessels is a key step in cells entering the tissues, the lymphatic vessels also serve as a conduit for cells that are recruited and localized through chemoattractant gradients. Furthermore, the growth and remodeling of lymphatic vessels in pathologies is influenced by chemokines and their receptors expressed by lymphatic endothelial cells (LECs) in and around the pathological tissue. In this review we summarize the diverse role played by specific chemokines and their receptors in shaping the interaction of lymphatic vessels, immune cells, and other pathological cell types in physiology and disease