Identification of novel neutralizing single-chain antibodies against vascular endothelial growth factor receptor 2

Human vascular endothelial growth factor (VEGF) and its receptor (VEGFR-2/kinase domain receptor [KDR]) play a crucial role in angiogenesis, which makes the VEGFR-2 signaling pathway a major target for therapeutic applications. In this study, a single-chain antibody phage display library was constructed from spleen cells of mice immunized with recombinant human soluble extracellular VEGFR-2/KDR consisting of all seven extracellular domains (sKDR D1-7) to obtain antibodies that block VEGF binding to VEGFR-2. Two specific single-chain antibodies (KDR1.3 and KDR2.6) that recognized human VEGFR-2 were selected; diversity analysis of the clones was performed by BstNI fingerprinting and nucleotide sequencing. The single-chain variable fragments (scFvs) were expressed in soluble form and specificity of interactions between affinity purified scFvs and VEGFR-2 was confirmed by ELISA. Binding of the recombinant antibodies for VEGFR-2 receptors was investigated by surface plasmon resonance spectroscopy. In vitro cell culture assays showed that KDR1.3 and KDR2.6 scFvs significantly suppressed the mitogenic response of human umbilical vein endothelial cells to recombinant human VEGF 165 in a dose-dependent manner, and reduced VEGF-dependent cell proliferation by 60% and 40%, respectively. In vivo analysis of these recombinant antibodies in a rat cornea angiogenesis model revealed that both antibodies suppressed the development of new corneal vessels (p < 0.05). Overall, in vitro and in vivo results disclose strong interactions of KDR1.3 and KDR2.6 scFvs with VEGFR-2. These findings indicate that KDR1.3 and KDR2.6 scFvs are promising antiangiogenic therapeutic agents. © 2011 International Union of Biochemistry and Molecular Biology, Inc

Understanding the potential in vitro modes of action of bis(β‐diketonato) oxovanadium(IV) complexes

To understand the potential in vitro modes of action of bis(β-diketonato) oxovanadium(IV) complexes, nine compounds of varying functionality have been screened using a range of biological techniques. The antiproliferative activity against a range of cancerous and normal cell lines has been determined, and show these complexes are particularly sensitive against the lung carcinoma cell line, A549. Annexin V (apoptosis) and Caspase-3/7 assays were studied to confirm these complexes induce programmed cell death. While gel electrophoresis was used to determine DNA cleavage activity and production of reactive oxygen species (ROS), the Comet assay was used to determine induced genomic DNA damage. Additionally, Förster resonance energy transfer (FRET)-based DNA melting and fluorescent intercalation displacement assays have been used to determine the interaction of the complexes with double strand (DS) DNA and to establish preferential DNA base-pair binding (AT versus GC)

Evaluation of the oblique fissure on lateral chest radiographs in coal workers pneumoconiosis

Objective: The oblique fissure separates the upper lobe from the lower lobe in the left lung and the upper lobe from the lower and middle lobes in the right lung. The aim of the present study was to compare the projection of the oblique fissure observed on lateral chest radiographs of retired coal mine workers who had coal worker's pneumoconiosis (CWP) with that of healthy men. Materials and methods: The study included 120 men divided into three groups: retired coal miner patients diagnosed with CWP (group 1), men ?38 years of age with no CWP (group 2, first control group), and men ?37 years of age with no CWP (group 3, second control group). The projection of the oblique fissure observed on lateral chest radiographs (left side) were evaluated using geometric morphometrics. Geometric morphometric analysis of each sample was performed using 11 pre-specified landmarks designated on X-ray images, using tpsDig2 software. A text file containing the landmark coordinates of the 120 samples was created, and the data in the text file was statistically analyzed using Morpheus software. Results: The projection of the oblique fissure differed significantly between the CWP patients and the control groups (p<0.05). Conclusion: We suggest that the difference in the projection of the oblique fissure between men with and without CWP was attributable to a protrusion in the fissure caused by nodules that developed in the lungs of coal miners after years of dust inhalation

Evaluation of the oblique fissure on lateral chest radiographs in coal workers pneumoconiosis

WOS: 000321023200013PubMed ID: 23731051Objective: The oblique fissure separates the upper lobe from the lower lobe in the left lung and the upper lobe from the lower and middle lobes in the right lung. The aim of the present study was to compare the projection of the oblique fissure observed on lateral chest radiographs of retired coal mine workers who had coal worker's pneumoconiosis (CWP) with that of healthy men. Materials and methods: The study included 120 men divided into three groups: retired coal miner patients diagnosed with CWP (group 1), men >= 38 years of age with no CWP (group 2, first control group), and men 537 years of age with no CWP (group 3, second control group). The projection of the oblique fissure observed on lateral chest radiographs (left side) were evaluated using geometric morphometrics. Geometric morphometric analysis of each sample was performed using 11 pre-specified landmarks designated on X-ray images, using tpsDig2 software. A text file containing the landmark coordinates of the 120 samples was created, and the data in the text file was statistically analyzed using Morpheus software. Results: The projection of the oblique fissure differed significantly between the CWP patients and the control groups (p<0.05). Conclusion: We suggest that the difference in the projection of the oblique fissure between men with and without CWP was attributable to a protrusion in the fissure caused by nodules that developed in the lungs of coal miners after years of dust inhalation (Tab. 4, Fig. 4, Ref. 30). Full Text in PDF www.elis.sk

Policies for promoting entrepreneurship as a means of increasing social inclusion

The aim of this chapter is to show how an understanding of the policymaking process might lead to better formulation and outcomes of policies to support the social inclusion of minority and disadvantaged groups entrepreneurship. The chapter reviews the mixed evidence to date in relation to both enterprise policies more broadly and inclusive policies in particular and it proposes an alternative conceptualisation of the policy process that makes evident the need to take context more seriously. The chapter offers some pointers about the ways in which the needs and interests of minority entrepreneurs can be connected more clearly to the governmental agendas and the policy formulation that takes place at the governmental level