Designed antitumor peptide for targeted siRNA delivery into cancer spheroids

Antimicrobial/anticancer peptides (AMPs/ACPs) have shown promising results as new therapeutic agents in cancer therapy. Among them, the designed amphiphilic α-helical peptide G(IIKK)3I-NH2 (G3) displayed great affinity and specificity in targeting cancer cells. Here, we report new insights on how G3 penetrates cancer cells. G3 showed high specificity to HCT-116 colon cancer cells compared to the HDFs (human neonatal primary dermal fibroblasts) control. With high concentrations of peptide, a clear cancer cell membrane disruption was observed through SEM. Gene knockdown of the endocytic pathways demonstrated that an energy-dependent endocytic pathway is required for the uptake of the peptide. In addition, G3 can protect and selectively deliver siRNAs into cancer cells and successfully modulated their gene expression. Gene delivery was also tested in 3D cancer spheroids and showed deep penetration delivery into the cancer spheroids. Finally, the in vivo toxicity of G3 was evaluated on zebrafish embryos, showing an increasing toxicity effect with concentration. However, the toxicity of the peptide was attenuated when complexed with siRNA. In addition, negligible toxicity was observed at the concentration range for efficient gene delivery. The current results demonstrate that G3 is promising as an excellent agent for cancer therapy

INNOVATIVE PROCUREMENT METHODS IN RAIL TRANSIT PROJECTS: BALTIMORE\u27S TURNKEY EXPERIENCE

The Federal Transit Administration\u27s (FTA\u27s) Turnkey Demonstration Program, as mandated by the Intermodal Surface Transportation Efficiency Act of 1991, is required to select a number of prototype projects for comparison of turnkey to traditional procurement methods. The Baltimore Central Light Rail Line (CLRL) Phase 2 Extensions project is one of four prototype projects currently selected for comparison. In 1992, the Maryland Transit Administration (MTA) started service on Phase 1 of the CLRL project--a 36.2-km-long system with 25 station stops, procured under the traditional method (that is, design/bid/build). The CLRL Phase 2 Extensions project, procured under the turnkey method, completed the 48.3-km plan with three extensions to the original 36.2 km. FTA has requested an evaluation of the effectiveness of the turnkey procurement method in advancing the design, construction, testing, start-up, and initial operation of the CLRL Phase 2 Extensions project. This evaluation effort focuses on the procurement phase of the project. As part of this evaluation, the contractual differences between the conventional contracting method used in Phase 2 and the design/build, or turnkey, contracting mechanisms and incentives that were used in Phase 2 Extensions project are discussed. The focus of this discussion is on the comparative differences of the specific contracting mechanisms and incentives used by MTA in the procurement phase of the two comparable projects, CLRL Phases 1 and 2. This discussion also examines the effectiveness of the procurement approach implemented by MTA as compared with other turnkey contracts

What Determines Soft Costs in Major Public Transit Fixed Guideway Projects? Examining Four Decades of Experience

Soft costs in major public transportation fixed guideway projects generally account for about 20% of the project\u27s capital cost, but wide variation in the relative magnitude of soft costs can make them a major factor in the public deliberation over whether to build a transit project. In addition, soft costs are often poorly understood and have varied considerably more than industry estimation techniques predicted. This paper examines the actual capital expenditures from 59 U.S. light and heavy rail projects and seeks to explain the wide variance in soft costs (from 11% to 54% of construction costs) by analyzing project characteristics such as complexity, physical attributes, sponsor agency traits, and the circumstances of project development. The analysis develops a multivariate regression establishing a cost relationship between 10 project characteristics and resulting soft costs as a percentage of hard (construction) costs. These results can be used to make more accurate estimates of soft costs for future projects