Suppression of anchorage-independent growth and matrigel invasion and delayed tumor formation by elevated expression of fibulin-1D in human fibrosarcoma-derived cell lines

Using differential display, we identified an mRNA that is markedly down-regulated in cell line 6A/SB1, derived from a fibrosarcoma formed in an athymic mouse following injection of carcinogen-transformed MSU-1.1 cells. The nontumorigenic parental cell strain, MSU-1.1, expresses high levels of this mRNA. Sequencing of the corresponding cDNA fragment revealed that it corresponded to an expressed sequence tag, which ultimately led to its identification as the fibulin-1D gene. Fibulin-1 is a cysteine-rich, calcium-binding extracellular matrix and plasma protein, which has four isoforms, A-D, derived from alternative splicing. Northern and Western blotting analysis of 16 cell lines established from tumors formed in athymic mice by MSU-1.1-derived cell strains independently transformed in culture showed that 44% exhibited low level or lack of expression of fibulin-1D mRNA and protein. In a similar analysis of 15 malignant cell lines derived from patients, 80% showed low level or no expression. To study the role of fibulin-1D in transformation, we transfected 6A/SB1 cells and a human fibrosarcoma-derived cell line (SHAC) with a fibulin-1D cDNA expression construct. Transfectants displaying high levels of fibulin-1D were isolated and characterized. Elevated expression of fibulin-1D led to reduced ability to form colonies in soft agar and reduced invasive potential as tested in a matrigel in vitro invasion assay. Furthermore, expression of fibulin-1D resulted in a markedly extended latency in tumor formation in athymic mice. These results indicate that low expression of fibulin-1D plays a role in tumor formation and invasion.     

Stabilization loop of a two axes gimbal system using self-tuning PID type fuzzy controller

The application of inertial stabilization system is to stabilize the sensor's line of sight toward a target by isolating the sensor from the disturbances induced by the operating environment. The aim of this paper is to present two axes gimbal system. The gimbals torque relationships are derived using Lagrange equation considering the base angular motion and dynamic mass unbalance. The stabilization loops are constructed with cross coupling unit utilizing proposed fuzzy PID type controller. The overall control system is simulated and validated using MATLAB. Then, the performance of proposed controller is evaluated comparing with conventional PI controller in terms of transient response analysis and quantitative study of error analysis. The simulation results obtained in different conditions prove the efficiency of the proposed fuzzy controller which offers a better response than the classical one, and improves further the transient and steady-state performance.     

Hepatocyte growth factor-stimulated renal tubular mitogenesis: effects on expression of c-myc, c-fos, c-met, VEGF and the VHL tumour-suppressor and related genes

Hepatocyte growth factor (HGF/SF) is a potent renal proximal tubular cell (PTEC) mitogen involved in renal development. HGF/SF is the functional ligand for the c-met proto-oncogene, and germline c-met mutations are associated with familial papillary renal cell carcinoma. Somatic von Hippel-Lindau disease tumour-suppressor gene (VHL) mutations are frequently detected in sporadic clear cell renal cell carcinomas (RCC), and germline VHL mutations are the commonest cause of familial clear cell RCC. pVHL binds to the positive regulatory components of the trimeric elongin (SIII) complex (elongins B and C) and has been observed to deregulate expression of the vascular endothelial growth factor (VEGF) gene. HGF/SF has similarly been reported to up-regulate expression of the VEGF gene in non-renal experimental systems. To investigate the mechanism of HGF/SF action in PTECs and, specifically, to examine potential interactions between the HGF/c-met and the VHL-mediated pathways for renal tubular growth control, we have isolated untransformed PTECs from normal kidneys, developed conditions for their culture in vitro and used these cells to investigate changes in mRNA levels of the VHL, elongin A, B and C, VEGF, c-myc, c-fos and c-met genes after HGF/SF exposure. Significant elevations in the mRNA levels of VEGF, c-myc, c-fos, c-met and elongins A, B and C, but not VHL, were detected after HGF/SF stimulation of human PTECs (P < 0.02), with a consistent order of peak levels observed over successive replicates (c-fos at 1 h, VEGF at 2-4 h, c-myc, at 4 h, followed by c-met and all three elongin subunits at 8 h). This study highlights the spectrum of changes in gene expression observed in PTECs after HGF/SF stimulation and has identified possible candidate mediators of the HGF/SF-induced mitogenic response. Our evidence would suggest that the changes in PTEC VEGF expression induced by HGF/SF are mediated by a VHL-independent pathway.     

SOS–A sample ordering system for delivering “assay-ready” compound plates for drug screening

Many bottlenecks in drug discovery have been addressed with the advent of new assay and instrument technologies. However, storing and processing chemical compounds for screening remains a challenge for many drug discovery laboratories. Although automated storage and retrieval systems are commercially available for medium to large collections of chemical samples, these samples are usually stored at a central site and are not readily accessible to satellite research labs.Drug discovery relies on the rapid testing of new chemical compounds in relevant biological assays. Therefore, newly synthesized compounds must be readily available in various formats to biologists performing screening assays. Until recently, our compounds were distributed in screw cap vials to assayists who would then manually transfer and dilute each sample in an “assay-ready” compound plate for screening. The vials would then be managed by the individuals in an ad hoc manner.To relieve the assayist from searching for compounds and preparing their own assay-ready compound plates, a newly customized compound storage system with an ordering software application was implemented at our research facility that eliminates these bottlenecks. The system stores and retrieves compounds in 1 mL-mini-tubes or microtiter plates, facilitates compound searching by identifier or structure, orders compounds at varying concentrations in specified wells on 96- or 384-well plates, requests the addition of controls (vehicle or reference compounds), etc. The orders are automatically processed and delivered to the assayist the following day for screening. An overview of our system will demonstrate that we minimize compound waste and ensure compound integrity and availability.     

EXTRACTION OF TRANSITION METALS FROM WOOD PULP FIBERS USING SUPERCRITICAL CARBON DIOXIDE

The supercritical fluid extraction of manganese and iron from solid matrix (wood pulp) is demonstrated experimentally. Supercritical carbon diodixe is used together with lithium bis(trifluoroethyl) dithiocarbonate (FDDC) as the ion-pairing reagent. The metal content of the pulp samples was determined by inductively couple plasma atomic emission spectroscopy (ICP-AES). Optimum conditions were determined to be 30 min each of static and dynamic extraction with 200 atm pressure at 40 °C. Thermomechanical pulp (TMP) extracted under these parameters exhibits an 80% decrease in manganese content, and 91% after a second, sequential extraction. The simultaneous removal of iron was found to be only 6% and supports evidence that it is much more strongly bound to the TMP matrix than manganese.     

Provisioning Methods for Bit-Rate-Differentiated Services in Hybrid Optical Networks

This paper addresses the problem of routing and wavelength assignment of bit-rate-differentiated optical services in a hybrid network. Hybrid optical networks are composed of resources, such as fiber links and photonic/electronic switches, that vary in their capabilities and transmission qualities. These networks are also responsible for the realization of optical services with varying quality-of-service (QoS) guarantees. In such networks, it is required to have a cost-effective assignment of the optical and electronic resources to these services in order to maximize the revenue of the network operator. This paper deals with optical services that are defined according to their tolerance to transmission impairments. We first divide the provisioning problem into two phases: (1) routing and (2) wavelength assignment and regeneration reservation. In the routing phase, a set of k-routes are generated to select from in the second phase, where each route optimizes a specific aspect of the problem (e.g., number of hops, maximum accumulated noise, etc.). The second phase, using the information about the resources along each route, attempts at finding the best wavelength allocation on that route such that the signal quality meets the service-level agreement (SLA). The second phase also uses the minimum number of regenerator ports on intermediate nodes for the purpose of wavelength translation and signal clean-up. Comparisons of the above scheme with a probing-based method, reveal substantial enhancements to the blocking performance with a maximum running time increase of 60%. In addition, the use of multiple routes provides higher reduction in the blocking probability over single-routing schemes. Moreover, the proposed, non-pessimistic, provisioning approach has a major impact on reducing the regeneration budget of the network.     

Design and Expected Performance of GISMO-2, a Two Color Millimeter Camera for the IRAM 30 m Telescope

We present the main design features for the GISMO-2 bolometer camera, which we build for background-limited operation at the IRAM 30 m telescope on Pico Veleta, Spain. GISMO-2 will operate simultaneously in the 1 and 2 mm atmospherical windows. The 1 mm channel uses a \(32 \times 40\) TES-based backshort under grid (BUG) bolometer array, the 2 mm channel operates with a \(16 \times 16\) BUG array. The camera utilizes almost the entire full field of view provided by the telescope. The optical design of GISMO-2 was strongly influenced by our experience with the GISMO 2 mm bolometer camera, which is successfully operating at the 30 m telescope. GISMO is accessible to the astronomical community through the regular IRAM call for proposals.