p53 status correlates with histopathological response in patients with soft tissue sarcomas treated using isolated limb perfusion with TNF-alpha and melphalan.

BACKGROUND: Recombinant tumor necrosis factor-alpha (TNF-alpha) combined to melphalan is clinically administered through isolated limb perfusion (ILP) for regionally advanced soft tissue sarcomas of the limbs. In preclinical studies, wild-type p53 gene is involved in the regulation of cytotoxic action of TNF-alpha and loss of p53 function contributes to the resistance of tumour cells to TNF-alpha. The relationship between p53 status and response to TNF-alpha and melphalan in patients undergoing ILP is unknown. PATIENTS AND METHODS: We studied 110 cases of unresectable limbs sarcomas treated by ILP. Immunohistochemistry was carried out using DO7mAb, which reacts with an antigenic determinant from the N-terminal region of both the wild-type and mutant forms of the p53 protein, and PAb1620mAb, which reacts with the 1620 epitope characteristic of the wild-type native conformation of the p53 protein. The immunohistochemistry data were then correlated with various clinical parameters. RESULTS: P53DO7 was found expressed at high levels in 28 patients, whereas PAb1620 was negative in 20. The tumours with poor histological response to ILP with TNF-alpha and melphalan showed significantly higher levels of p53-mutated protein. CONCLUSIONS: Our results might be a clue to a role of p53 protein status in TNF-alpha and melphalan response in clinical use.

Thin Maintenance Surfaces - Phase II, TR-435, 2003

In recent years there has been renewed interest in using preventive maintenance techniques to extend pavement life and to ensure low life cycle costs for our road infrastructure network. Thin maintenance surfaces can be an important part of a preventive maintenance program for asphalt cement concrete roads. The Iowa Highway Research Board has sponsored Phase Two of this research project to demonstrate the use of thin maintenance surfaces in Iowa and to develop guidelines for thin maintenance surface uses that are specific to Iowa. This report documents the results of test section construction and monitoring started in Phase One and continued in Phase Two. The report provides a recommended seal coat design process based on the McLeod method and guidance on seal coat aggregates and binders. An update on the use of local aggregates for micro-surfacing in Iowa is included. Winter maintenance guidelines for thin maintenance surfaces are reported herein. Finally, Phase One's interim, qualitative thin maintenance surface guidelines are supplemented with Phase two's revised, quantitative guidelines. When thin maintenance surfaces are properly selected and applied, they can improve the pavement surface condition index and the skid resistance of pavements. For success to occur, several requirements must be met, including proper material selection, design, application rate, workmanship, and material compatibility, as well as favorable weather during application and curing. Specific guidance and recommendations for many types of thin maintenance surfaces and conditions are included in the report.

Peroxisome proliferator-activated receptor mediates cross-talk with thyroid hormone receptor by competition for retinoid X receptor. Possible role of a leucine zipper-like heptad repeat.

The peroxisome proliferator-activated receptors (PPAR) and thyroid hormone receptors (TR) are members of the nuclear receptor superfamily, which regulate lipid metabolism and tissue differentiation. In order to bind to DNA and activate transcription, PPAR requires the formation of heterodimers with the retinoid X receptor (RXR). In addition to activating transcription through its own response elements, PPAR is able to selectively down-regulate the transcriptional activity of TR, but not vitamin D receptor. The molecular basis of this functional interaction has not been fully elucidated. By means of site-directed mutagenesis of hPPAR alpha we mapped its inhibitory action on TR to a leucine zipper-like motif in the ligand binding domain of PPAR, which is highly conserved among all subtypes of this receptor and mediates heterodimerization with RXR. Replacement of a single leucine by arginine at position 433 of hPPAR alpha (L433R) abolished heterodimerization of PPAR with RXR and consequently its trans-activating capacity. However, a similar mutation of a leucine residue to arginine at position 422 showed no alteration of heterodimerization, DNA binding, or transcriptional activation. The dimerization deficient mutant L433R was no longer able to inhibit TR action, demonstrating that the selective inhibitory effect of PPAR results from the competition for RXR as well as possibly for other TR-auxiliary proteins. In contrast, abolition of DNA binding by a mutation in the P-box of PPAR (C122S) did not eliminate the inhibition of TR trans-activation, indicating that competition for DNA binding is not involved. Additionally, no evidence for the formation of PPAR:TR heterodimers was found in co-immunoprecipitation experiments. In summary, we have demonstrated that PPAR selectively inhibits the transcriptional activity of TRs by competition for RXR and possibly non-RXR TR-auxiliary proteins. In contrast, this functional interaction is independent of the formation of PPAR:TR heterodimers or competition for DNA binding.

SDF 1-alpha (CXCL12) triggers glutamate exocytosis from astrocytes on a millisecond time scale: Imaging analysis at the single-vesicle level with TIRF microscopy

Chemokines are small chemotactic molecules widely expressed throughout the central nervous system. A number of papers, during the past few years, have suggested that they have physiological functions in addition to their roles in neuroinflammatory diseases. In this context, the best evidence concerns the CXC-chemokine stromal cell-derived factor (SDF-1alpha or CXCL12) and its receptor CXCR4, whose signalling cascade is also implicated in the glutamate release process from astrocytes. Recently, astrocytic synaptic like microvesicles (SLMVs) that express vesicular glutamate transporters (VGLUTs) and are able to release glutamate by Ca(2+)-dependent regulated exocytosis, have been described both in tissue and in cultured astrocytes. Here, in order to elucidate whether SDF-1alpha/CXCR4 system can participate to the brain fast communication systems, we investigated whether the activation of CXCR4 receptor triggers glutamate exocytosis in astrocytes. By using total internal reflection (TIRF) microscopy and the membrane-fluorescent styryl dye FM4-64, we adapted an imaging methodology recently developed to measure exocytosis and recycling in synaptic terminals, and monitored the CXCR4-mediated exocytosis of SLMVs in astrocytes. We analyzed the co-localization of VGLUT with the FM dye at single-vesicle level, and observed the kinetics of the FM dye release during single fusion events. We found that the activation of CXCR4 receptors triggered a burst of exocytosis on a millisecond time scale that involved the release of Ca(2+) from internal stores. These results support the idea that astrocytes can respond to external stimuli and communicate with the neighboring cells via fast release of glutamate.

A polymorphic microsatellite in the tumor necrosis factor alpha promoter identifies an allele unique to the NZW mouse strain.

We have amplified a (CA)n:(GT)n microsatellite from the TNF promoters of a panel of mouse strains using the polymerase chain reaction. The length of the microsatellites was polymorphic, with eight alleles observed among 15 inbred strains bearing seven distinct H-2 haplotypes, and four outbred strains. In B10 congenic strains, the TNF allele detected by microsatellite polymorphism segregated with the MHC, and in recombinant haplotypes (NOD, NZW), it segregated with H-2D. The TNF allele found in the NZW strain (H-2z) was distinct from those of all other haplotypes, consistent with the hypothesis that this strain may carry a genetic defect in TNF production.

Two Cases of Interferon-Alpha-Induced Sarcoidosis Koebnerized along Venous Drainage Lines: New Pathogenic Insights and Review of the Literature of Interferon-Induced Sarcoidosis.

Sarcoidosis is a systemic granulomatous disorder of unknown origin commonly affecting the lung, the lymphoid system and the skin. We report here two cases of cutaneous sarcoidosis in two former intravenous drug users following interferon (IFN)-α and ribavirin therapy for chronic hepatitis C. Both patients developed skin sarcoidosis along venous drainage lines of both forearms, coinciding with the areas of prior drug injections. The unique distribution of the skin lesions suggests that tissue damage induced by repeated percutaneous drug injections represents a trigger for the local skin manifestation of sarcoidosis. Interestingly, skin damage was recently found to induce the local expression IFN-α, a well-known trigger of sarcoidosis in predisposed individuals. Here we review the literature on sarcoidosis elicited in the context of IFN-α therapy and propose a new link between the endogenous expression of IFN-α and the induction of disease manifestations in injured skin. © 2013 S. Karger AG, Basel.

A role for the alpha-chain connecting peptide motif in mediating TCR-CD8 cooperation.

To generate peripheral T cells that are both self-MHC restricted and self-MHC tolerant, thymocytes are subjected to positive and negative selection. How the TCR discriminates between positive and negative selection ligands is not well understood, although there is substantial evidence that the CD4 and CD8 coreceptors play an important role in this cell fate decision. We have previously identified an evolutionarily conserved motif in the TCR, the alpha-chain connecting peptide motif (alpha-CPM), which allows the TCR to deliver positive selection signals. Thymocytes expressing alpha-CPM-deficient receptors do not undergo positive selection, whereas their negative selection is not impaired. In this work we studied the ligand binding and receptor function of alpha-CPM-deficient TCRs by generating T cell hybridomas expressing wild-type or alpha-CPM-deficient forms of the T1 TCR. This K(d)-restricted TCR is specific for a photoreactive derivative of the Plasmodium berghei circumsporozoite peptide(252-260) IASA-YIPSAEK(ABA)I and is therefore amenable to TCR photoaffinity labeling. The experiments presented in this work show that alpha-CPM-deficient TCRs fail to cooperate with CD8 to enhance ligand binding and functional responses.

Development of an Improved Agricultural-Based Deicing Product, TR-581, 2010

Snow and ice removal on public streets is a critical part of the work of departments of transportation in northern U.S. states, including Iowa. Iowa is also a state rich in agricultural resources, some of which undergo industrial processes that generate a number of byproducts, e.g., in converting corn to ethanol or soy to biodiesel. It would be desirable to find those that, with a minimum of additional processing, can be used as a deicing compound, either alone or in combination with products currently in use. The focus of this work is therefore to investigate by-products from agricultural processes that may be suitable for use as deicing applications. This topic has been investigated in the past by others, with many patented products described in the literature. An initial screening was carried out to assess the potential acceptability of selected commercial products, as well as a glycerol developed for this project. Based on the variety of parameters tested, the product combination that shows the greatest promise for future application consists of 80% glycerol with 20% NaCl.

Hypothyroid myopathy as a complication of interferon alpha therapy for chronic hepatitis C virus infection.

Interferon alpha (IFN-alpha) therapy is associated with a number of immunological side-effects, including autoimmune diseases and a 10% prevalence of thyroiditis. Hepatitis C virus (HCV) infection itself predisposes to autoimmune phenomena including hypothyroidism and myositis. The development of clinical hypothyroidism in the presence of positive thyroid antibodies in patients infected with HCV and treated with IFN-alpha suggests a possible association between the viral disease and the therapy. HCV infection may predispose to autoimmune thyroid disease and IFN-alpha therapy may secondarily lead to the development of thyroid dysfunctions. We report the single case of a female patient who developed a severe proximal myopathy in conjunction with primary hypothyroidism (Hoffmann's syndrome) secondarily to IFN-alpha therapy for HCV infection. This case highlights the need for careful clinical and biological monitoring for potential side-effects in such patients.

Embankment Quality Phase II Final Report, TR-401, 1999

Phase II research included the following: (1) develop and evaluate alternative soil design and embankment construction specifications based on soil type, moisture, density, stability, and compaction process; (2) assess various quality control and acceptance procedures with a variety of in-situ test methods including the Dual-mass Dynamic Cone Penetrometer (DCP); and (3) develop and design rapid field soil identification methods. At the start of the research, soils were divided into cohesive and cohesionless soil types, with each category being addressed separately. Cohesionless soils were designated as having less than 36% fines content (material passing the No. 200 sieve) and cohesive soils as having greater than 36% fines content. Subsequently, soil categories were refined based not only on fines content but soil plasticity as well. Research activities included observations of fill placement, in-place moisture and density testing, and dual-mass DCP index testing on several highway embankment projects throughout Iowa. Experiments involving rubber-tired and vibratory compaction, lift thickness changes, and disk aeration were carried out for the full range of Iowa soils. By testing for soil stability the DCP was found to be a valuable field tool for quality control, whereby shortcomings from density testing (density gradients) were avoided. Furthermore, critical DCP index values were established based on soil type and compaction moisture content.

Access Management Awareness Program Phase II Report, TR-402, 1997

This report presents the results of a number of detailed Iowa access management case studies. Case studies were selected to provide a cross-section of locations and community sizes in Iowa as well as a variety of project types. Generally, access management projects completed during the mid-1990s were chosen as case studies. Projects ranging from driveway consolidation to full raised medians were analyzed on a before and after basis in terms of traffic safety, traffic operations, and adjacent business vitality. Sources of information used for the case study analysis included: road project files; traffic accident records; state sales tax records; and personal interviews of business owners, business customers, and local officials. The case study results from Iowa essentially confirm results of previous access management research from around the nation. Recent access in Iowa had significant, positive impacts in terms of traffic safety. The average reduction of annual accidents and accident rates on improved roadways was approximately 40%. Improvements in access management also led to significantly better roadway operations for most case studies. Although a small number of individual businesses do report sales losses and/or customer complaints once projects have been completed, access management projects in Iowa have not had an adverse impact on the majority of businesses located along them. In fact, some access management projects in Iowa seem to have contributed to an improved business environment along the corridors that have been improved. The results from the Iowa case studies presented in this report will be used to develop access management education materials for Iowa transportation professionals and other audiences interested in the impacts of access management.

Performance Based Specifications, TR-403, 1998

The goal in highway construction and operation has shifted from method based specifications to specifications relating desired performance attributes to materials, mix designs, and construction methods. Shifting from method specifications to performance based specifications can work as an incentive or disincentive for the contractor to improve performance or extend pavement life. This literature search was directed at a review of existing portland cement concrete performance specification development, and the criteria that can effectively measure pavement performance. The criteria identified in the literature include concrete strength, slab thickness, air content, initial smoothness, water-cement ratio, unit weight, and slump. A description of each criterion, along with the advantages, disadvantages, and test methods for each are identified. Also included are the results from a survey that was sent out to various state, federal, and trade agencies. The responses indicated that 53% currently use or are developing a performance based specification program. Of the 47% of agencies that do not use a performance based specification program, over 34% indicated that they would consider a similar program. The most commonly measured characteristics include thickness, strength, smoothness, and air content. Lastly recommendations and conclusions are made regarding other factors that affect pavement performance and a proposed second phase of the research is suggested. The research team suggests that a regional expert task group be formed to identify performance levels and criteria. The results of that effort will guide the research team in the development of new or revised specifications.

Maximizing the Use of Roadway Weather Information Systems: Final Report, TR-404, 1998

This project was proposed as Phase I of a 2-phase program to evaluate the present use of weather information by Iowa Department of Transportation (IaDOT) personnel, recommend revised procedures, and then implement the resulting recommendations. Midway through Phase I (evaluation phase) the FORETELL project was funded. This project is a multi-state venture that engages the National Weather Service (NWS) and the Forecast Systems Laboratory of the National Oceanic and Atmospheric Administration and proposes to supplant the current weather information-generation and distribution system with an advanced system based on state-of-the-art technologies. The focus of the present project was therefore refined to consider use of weather data by IaDOT personnel, and the training programs needed to more effectively use these data. Results of the survey revealed that two major areas - training of personnel on use of data from whatever source and more precise information of frost formation - are not addressed in the FORETELL project. These aspects have been the focus of the present project.

Evaluation of Photoacoustic Spectroscopy for Quality Control of Cement, TR-409, 1998

This report discusses the feasibility of using infrared photoacoustic spectroscopy (PAS) as a viable technique that can quickly provide information on cement composition prior to use. The PAS technique is of interest because the cost is much lower than for other types of instrumentation used for mineral analysis, it requires virtually no sample preparation, and it can perform multi-component analysis in a matter of minutes. Feasibility of the technique was based on the ability of PAS to identify and quantify sulfate species and major cement matrix components. Strengths and limitations of the technique are presented.

Investigation of Two Bridge Alternatives for Low Volume Roads - Phase II Volume 2 of 2, Concept 2: Beam in Slab Bridge, TR-410, 2000

This project continues the research which addresses the numerous bridge problems on the Iowa secondary road system. It is a continuation (Phase 2) of Project HR-382, in which two replacement alternatives (Concept 1: Steel Beam Precast Units and Concept 2: Modification of the Benton County Beam-in-Slab Bridge) were investigated. In previous research for concept 1, a precast unit bridge was developed through laboratory testing. The steel-beam precast unit bridge requires the fabrication of precast double-tee (PCDT) units, each consisting of two steel beams connected by a reinforced concrete deck. The weight of each PCDT unit is minimized by limiting the deck thickness to 4 in., which permits the units to be constructed off-site and then transported to the bridge site. The number of units required is a function of the width of bridge desired. Once the PCDT units are connected, a cast-in-place reinforced concrete deck is cast over the PCDT units and the bridge railing attached. Since the steel beam PCDT unit bridge design is intended primarily for use on low-volume roads, used steel beams can be utilized for a significant cost savings. In previous research for concept 2, an alternate shear connector (ASC) was developed and subjected to static loading. In this investigation, the ASC was subjected to cyclic loading in both pushout specimens and composite beam tests. Based on these tests, the fatigue strength of the ASC was determined to be significantly greater than that required in typical low volume road single span bridges. Based upon the construction and service load testing, the steel-beam precast unit bridge was successfully shown to be a viable low volume road bridge alternative. The construction process utilized standard methods resulting in a simple system that can be completed with a limited staff. Results from the service load tests indicated adequate strength for all legal loads. An inspection of the bridge one year after its construction revealed no change in the bridge's performance. Each of the systems previously described are relatively easy to construct. Use of the ASC rather than the welded studs significantly simplified the work, equipment, and materials required to develop composite action between the steel beams and the concrete deck.