Coculture of bladder urothelial and smooth muscle cells on small intestinal submucosa: potential applications for tissue engineering technology.

PURPOSE: Small intestinal submucosa is a xenogenic, acellular, collagen rich membrane with inherent growth factors that has previously been shown to promote in vivo bladder regeneration. We evaluate in vitro use of small intestinal submucosa to support the individual and combined growth of bladder urothelial cells and smooth muscle cells for potential use in tissue engineering techniques, and in vitro study of the cellular mechanisms involved in bladder regeneration. MATERIALS AND METHODS: Primary cultures of human bladder urothelial cells and smooth muscle cells were established using standard enzymatic digestion or explant techniques. Cultured cells were then seeded on small intestinal submucosa at a density of 1 x 105 cells per cm.2, incubated and harvested at 3, 7, 14 and 28 days. The 5 separate culture methods evaluated were urothelial cells seeded alone on the mucosal surface of small intestinal submucosa, smooth muscle cells seeded alone on the mucosal surface, layered coculture of smooth muscle cells seeded on the mucosal surface followed by urothelial cells 1 hour later, sandwich coculture of smooth muscle cells seeded on the serosal surface followed by seeding of urothelial cells on the mucosal surface 24 hours later, and mixed coculture of urothelial cells and smooth muscle cells mixed and seeded together on the mucosal surface. Following harvesting at the designated time points small intestinal submucosa cell constructs were formalin fixed and processed for routine histology including Masson trichrome staining. Specific cell growth characteristics were studied with particular attention to cell morphology, cell proliferation and layering, cell sorting, presence of a pseudostratified urothelium and matrix penetrance. To aid in the identification of smooth muscle cells and urothelial cells in the coculture groups, immunohistochemical analysis was performed with antibodies to alpha-smooth muscle actin and cytokeratins AE1/AE3. RESULTS: Progressive 3-dimensional growth of urothelial cells and smooth muscle cells occurred in vitro on small intestinal submucosa. When seeded alone urothelial cells and smooth muscle cells grew in several layers with minimal to no matrix penetration. In contrast, layered, mixed and sandwich coculture methods demonstrated significant enhancement of smooth muscle cell penetration of the membrane. The layered and sandwich coculture techniques resulted in organized cell sorting, formation of a well-defined pseudostratified urothelium and multilayered smooth muscle cells with enhanced matrix penetration. With the mixed coculture technique there was no evidence of cell sorting although matrix penetrance by the smooth muscle cells was evident. Immunohistochemical studies demonstrated that urothelial cells and smooth muscle cells maintain the expression of the phenotypic markers of differentiation alpha-smooth muscle actin and cytokeratins AE1/AE3. CONCLUSIONS: Small intestinal submucosa supports the 3-dimensional growth of human bladder cells in vitro. Successful combined growth of bladder cells on small intestinal submucosa with different seeding techniques has important future clinical implications with respect to tissue engineering technology. The results of our study demonstrate that there are important smooth muscle cell-epithelial cell interactions involved in determining the type of in vitro cell growth that occurs on small intestinal submucosa. Small intestinal submucosa is a valuable tool for in vitro study of the cell-cell and cell-matrix interactions that are involved in regeneration and various disease processes of the bladder.

Temporary Traffic Control and Enforcement of Traffic Laws in Closed Road Sections, June 2007

Public travel by motor vehicles is often necessary in road and street sections that have been officially closed for construction, repair, and/or other reasons. This authorization is permitted in order to provide access to homes and businesses located beyond the point of closure. The MUTCD does address appropriate use of specific regulatory signs at the entrance to closed sections; however, direct guidance for temporary traffic control measures within these areas is not included but may be needed. Interpretation and enforcement of common practices may vary among transportation agencies. For example, some law enforcement officers in Iowa have indicated a concern regarding enforcement and jurisdiction of traffic laws in these areas because the Code of Iowa only appears to address violations on roadways open to “public travel.” Enforcement of traffic laws in closed road sections is desirable to maintain safety for workers and for specifically authorized road users. In addition, occasional unauthorized entry by motor vehicles is experienced in closed road areas causing property damage. Citations beyond simple trespass may be advisable to provide better security for construction sites, reduce economic losses from damage to completed work, and create safer work zones.

Application of Visualization and Simulation Program to Improve Work Zone Safety and Mobility, January 2010

A previous study sponsored by the Smart Work Zone Deployment Initiative, “Feasibility of Visualization and Simulation Applications to Improve Work Zone Safety and Mobility,” demonstrated the feasibility of combining readily available, inexpensive software programs, such as SketchUp and Google Earth, with standard two-dimensional civil engineering design programs, such as MicroStation, to create animations of construction work zones. The animations reflect changes in work zone configurations as the project progresses, representing an opportunity to visually present complex information to drivers, construction workers, agency personnel, and the general public. The purpose of this study is to continue the work from the previous study to determine the added value and resource demands created by including more complex data, specifically traffic volume, movement, and vehicle type. This report describes the changes that were made to the simulation, including incorporating additional data and converting the simulation from a desktop application to a web application.

Reclaimed Fly Ash as Select Fill Under PCC Pavement, November 1999

With the support of the Iowa Fly Ash Affiliates, research on reclaimed fly ash for use as a construction material has been ongoing since 1991. The material exhibits engineering properties similar to those of soft limestone or sandstone and a lightweight aggregate. It is unique in that it is rich in calcium, silica, and aluminum and exhibits pozzolanic properties (i.e. gains strength over time) when used untreated or when a calcium activator is added. Reclaimed Class C fly ashes have been successfully used as a base material on a variety of construction projects in southern and western Iowa. A pavement design guide has been developed with the support of the Iowa Fly Ash Affiliates. Soils in Iowa generally rate fair to poor as subgrade soils for paving projects. This is especially true in the southern quarter of the state and for many areas of eastern and western Iowa. Many of the soil types encountered for highway projects are unsuitable soils under the current Iowa DOT specifications. The bulk of the remaining soils are Class 10 soils. Select soils for use directly under the pavement are often difficult to find on a project, and in many instances are economically unavailable. This was the case for a 4.43-mile grading (STP-S- 90(22)-SE-90) and paving project in Wapello County. The project begins at the Alliant Utilities generating station in Chillicothe, Iowa, and runs west to the Monroe-Wapello county line. This road carries a significant amount of truck traffic hauling coal from the generating station to the Cargill corn processing plant in Eddyville, Iowa. The proposed 10-inch Portland Cement Concrete (PCC) pavement was for construction directly on a Class 10 soil subgrade, which is not a desirable condition if other alternatives are available. Wapello County Engineer Wendell Folkerts supported the use of reclaimed fly ash for a portion of the project. Construction of about three miles of the project was accomplished using 10 inches of reclaimed fly ash as a select fill beneath the PCC slab. The remaining mile was constructed according to the original design to be used as a control section for performance monitoring. The project was graded during the summers of 1998 and 1999. Paving was completed in the fall of 1999. This report presents the results of design considerations and laboratory and field testing results during construction. Recommendations for use of reclaimed fly ash as a select fill are also presented.

Handbook of Simplified Practice for Traffic Studies, November 2002

The Iowa Highway Research Board has identified the development of a simplified handbook of transportation studies as a high priority for the state of Iowa. The Center for Transportation Research and Education (CTRE) at Iowa State University was chosen to develop such a handbook. A well-executed, well-documented study is critical in the decision-making process for many transportation-related projects and in reporting to elected officials and members of the community. As more research is conducted in the area of transportation, study procedures in many cases have become more complex. It is often difficult for local jurisdictions with limited staff, training, experience, and time availability to perform these studies. The most commonly used publication for traffic studies is geared toward transportation professionals and professional engineers. That defining document, Manual of Transportation Studies (Institute of Transportation Engineers, 2000), is over 500 pages and includes several dozen types of transportation studies. Many of the transportation studies described in the manual are rarely (if ever) used by local jurisdictions. Further, those studies that are frequently used are at times very complex and possibly very costly to perform exactly as described. Local jurisdictions without the staff expertise to understand and apply the manual’s various studies have a need for a simplified handbook of procedures to perform common traffic studies themselves or properly define a scope of work to hire a consultant to perform the studies. This handbook describes simplified procedures that are easy to apply and are written for all potential users (civil engineers and traffic engineers, public works mangers, city managers and attorneys, and the general public).

Vehicle Traffic Map, 2010

Vehicle Traffic Map produced by the Iowa Department of Transportation.

Truck Traffic Map, 2010

Vehicle Traffic Map produced by the Iowa Department of Transportation.

Annual Average Daily Traffic on the Interstate System, 2010

Vehicle Traffic Map produced by the Iowa Department of Transportation.

Metabolic engineering of plants for the synthesis of polyhydroxyalkanoates

Synthesis of polyhydroxyalkanoates (PHAs) in crop is viewed as an attractive approach for the production of this family of biodegradable plastics in large quantities and at low costs. Synthesisof PHAs containing various monomers has so far been demonstrated in the cytosol, plastids, and peroxisomes of plants. Several biochemical pathways have been modifies to achieve this, including the isoprenois pathway, the fatty acid biosynthetic pathway, and the fatty acid

Endocytic and secretory traffic in Arabidopsis merge in the trans-Golgi network/early endosome, an independent and highly dynamic organelle.

Plants constantly adjust their repertoire of plasma membrane proteins that mediates transduction of environmental and developmental signals as well as transport of ions, nutrients, and hormones. The importance of regulated secretory and endocytic trafficking is becoming increasingly clear; however, our knowledge of the compartments and molecular machinery involved is still fragmentary. We used immunogold electron microscopy and confocal laser scanning microscopy to trace the route of cargo molecules, including the BRASSINOSTEROID INSENSITIVE1 receptor and the REQUIRES HIGH BORON1 boron exporter, throughout the plant endomembrane system. Our results provide evidence that both endocytic and secretory cargo pass through the trans-Golgi network/early endosome (TGN/EE) and demonstrate that cargo in late endosomes/multivesicular bodies is destined for vacuolar degradation. Moreover, using spinning disc microscopy, we show that TGN/EEs move independently and are only transiently associated with an individual Golgi stack.

Effects of traffic control on the soil physical quality and the cultivation of sugarcane

The cultivation of sugarcane with intensive use of machinery, especially for harvest, induces soil compaction, affecting the crop development. The control of agricultural traffic is an alternative of management in the sector, with a view to preserve the soil physical quality, resulting in increased sugarcane root growth, productivity and technological quality. The objective of this study was to evaluate the physical quality of an Oxisol with and without control traffic and the resulting effects on sugarcane root development, productivity and technological quality. The following managements were tested: no traffic control (NTC), traffic control consisting of an adjustment of the track width of the tractor and sugarcane trailer (TC1) and traffic control consisting of an adjustment of the track width of the tractor and trailer and use of an autopilot (TC2). Soil samples were collected (layers 0.00-0.10; 0.10-0.20 and 0.20-0.30 m) in the plant rows, inter-row center and seedbed region, 0.30 m away from the plant row. The productivity was measured with a specific weighing scale. The technological variables of sugarcane were measured in each plot. Soil cores were collected to analyze the root system. In TC2, the soil bulk density and compaction degree were lowest and total porosity and macroporosity highest in the plant row. Soil penetration resistance in the plant row, was less than 2 MPa in TC1 and TC2. Soil aggregation and total organic carbon did not differ between the management systems. The root surface and volume were increased in TC1 and TC2, with higher productivity and sugar yield than under NTC. The sugarcane variables did not differ between the managements. The soil physical quality in the plant row was preserved under management TC1 and TC2, with an improved root development and increases of 18.72 and 20.29 % in productivity and sugar yield, respectively.

Vehicular Traffic Flow Map, 1988

Traffic volumes represented on this map are annual average daily traffic volumes between major traffic generators: highway junctions and cities.

Vehicular Traffic Flow Map, 1990

Traffic volumes represented on this map are annual average daily traffic volumes between major traffic generators: highway junctions and cities.

Vehicular Traffic Flow Map, 1992

Traffic volumes represented on this map are annual average daily traffic volumes between major traffic generators: highway junctions and cities.