Probing the interaction between feline immunodeficiency virus and CD134 by using the novel monoclonal antibody 7D6 and the CD134 (Ox40) ligand.

The feline immunodeficiency virus (FIV) targets activated CD4-positive helper T cells preferentially, inducing an AIDS-like immunodeficiency in its natural host species, the domestic cat. The primary receptor for FIV is CD134, a member of the tumor necrosis factor receptor superfamily, and all primary viral strains tested to date use CD134 for infection. We examined the expression of CD134 in the cat using a novel anti-feline CD134 monoclonal antibody (MAb), 7D6, and showed that as in rats and humans, CD134 expression is restricted tightly to CD4+, and not CD8+, T cells, consistent with the selective targeting of these cells by FIV. However, FIV is also macrophage tropic, and in chronic infection the viral tropism broadens to include B cells and CD8+ T cells. Using 7D6, we revealed CD134 expression on a B220-positive (B-cell) population and on cultured macrophages but not peripheral blood monocytes. Moreover, macrophage CD134 expression and FIV infection were enhanced by activation in response to bacterial lipopolysaccharide. Consistent with CD134 expression on human and murine T cells, feline CD134 was abundant on mitogen-stimulated CD4+ T cells, with weaker expression on CD8+ T cells, concordant with the expansion of FIV into CD8+ T cells with progression of the infection. The interaction between FIV and CD134 was probed using MAb 7D6 and soluble CD134 ligand (CD134L), revealing strain-specific differences in sensitivity to both 7D6 and CD134L. Infection with isolates such as PPR and B2542 was inhibited well by both 7D6 and CD134L, suggesting a lower affinity of interaction. In contrast, GL8, CPG, and NCSU were relatively refractory to inhibition by both 7D6 and CD134L and, accordingly, may have a higher-affinity interaction with CD134, permitting infection of cells where CD134 levels are limiting.

Evaluation of Recycled Rubber in Asphalt Concrete, HR-330, 1996

The purpose of this research was to evaluate the performance and the use of asphalt rubber binders and recycled rubber granules in asphalt pavement in the state of Iowa. This five year research project was initiated in June 1991 and it was incorporated into Muscatine County Construction Project US 61 from Muscatine to Blue Grass over an existing 10 in. (25.4 cm) by 24 ft (7.3 m) jointed rigid concrete pavement constructed in 1957. The research site consisted of four experimental sections (one section containing rubber chip, one section containing reacted asphalt rubber in both binder and surface, and two sections containing reacted asphalt rubber in surface) and four control sections. This report contains findings of the University of Northern Iowa research team covering selected responsibilities of the research project "Determination of the aging and changing of the conventional asphalt binder and asphalt-rubber binder". Based on the laboratory test, the inclusion of recycled crumb rubber into asphalt affects the ductility of modified binder at various temperatures.

NFκB activation by modified vaccinia virus as a novel strategy to enhance neutrophil migration and HIV-specific T-cell responses.

Neutrophils are antigen-transporting cells that generate vaccinia virus (VACV)-specific T-cell responses, yet how VACV modulates neutrophil recruitment and its significance in the immune response are unknown. We generated an attenuated VACV strain that expresses HIV-1 clade C antigens but lacks three specific viral genes (A52R, K7R, and B15R). We found that these genes act together to inhibit the NFκB signaling pathway. Triple ablation in modified virus restored NFκB function in macrophages. After virus infection of mice, NFκB pathway activation led to expression of several cytokines/chemokines that increased the migration of neutrophil populations (Nα and Nβ) to the infection site. Nβ cells displayed features of antigen-presenting cells and activated virus-specific CD8 T cells. Enhanced neutrophil trafficking to the infection site correlated with an increased T-cell response to HIV vector-delivered antigens. These results identify a mechanism for poxvirus-induced immune response and alternatives for vaccine vector design.

Identification and characterization of novel classes of macrophage migration inhibitory factor (MIF) inhibitors with distinct mechanisms of action.

Macrophage migration inhibitory factor (MIF), a proinflammatory cytokine, is considered an attractive therapeutic target in multiple inflammatory and autoimmune disorders. In addition to its known biologic activities, MIF can also function as a tautomerase. Several small molecules have been reported to be effective inhibitors of MIF tautomerase activity in vitro. Herein we employed a robust activity-based assay to identify different classes of novel inhibitors of the catalytic and biological activities of MIF. Several novel chemical classes of inhibitors of the catalytic activity of MIF with IC(50) values in the range of 0.2-15.5 microm were identified and validated. The interaction site and mechanism of action of these inhibitors were defined using structure-activity studies and a battery of biochemical and biophysical methods. MIF inhibitors emerging from these studies could be divided into three categories based on their mechanism of action: 1) molecules that covalently modify the catalytic site at the N-terminal proline residue, Pro(1); 2) a novel class of catalytic site inhibitors; and finally 3) molecules that disrupt the trimeric structure of MIF. Importantly, all inhibitors demonstrated total inhibition of MIF-mediated glucocorticoid overriding and AKT phosphorylation, whereas ebselen, a trimer-disrupting inhibitor, additionally acted as a potent hyperagonist in MIF-mediated chemotactic migration. The identification of biologically active compounds with known toxicity, pharmacokinetic properties, and biological activities in vivo should accelerate the development of clinically relevant MIF inhibitors. Furthermore, the diversity of chemical structures and mechanisms of action of our inhibitors makes them ideal mechanistic probes for elucidating the structure-function relationships of MIF and to further determine the role of the oligomerization state and catalytic activity of MIF in regulating the function(s) of MIF in health and disease.

The G0/G1 switch gene 2 is a novel PPAR target gene.

PPARs (peroxisome-proliferator-activated receptors) alpha, beta/delta and gamma are a group of transcription factors that are involved in numerous processes, including lipid metabolism and adipogenesis. By comparing liver mRNAs of wild-type and PPARalpha-null mice using microarrays, a novel putative target gene of PPARalpha, G0S2 (G0/G1 switch gene 2), was identified. Hepatic expression of G0S2 was up-regulated by fasting and by the PPARalpha agonist Wy14643 in a PPARalpha-dependent manner. Surprisingly, the G0S2 mRNA level was highest in brown and white adipose tissue and was greatly up-regulated during mouse 3T3-L1 and human SGBS (Simpson-Golabi-Behmel syndrome) adipogenesis. Transactivation, gel shift and chromatin immunoprecipitation assays indicated that G0S2 is a direct PPARgamma and probable PPARalpha target gene with a functional PPRE (PPAR-responsive element) in its promoter. Up-regulation of G0S2 mRNA seemed to be specific for adipogenesis, and was not observed during osteogenesis or myogenesis. In 3T3-L1 fibroblasts, expression of G0S2 was associated with growth arrest, which is required for 3T3-L1 adipogenesis. Together, these data indicate that G0S2 is a novel target gene of PPARs that may be involved in adipocyte differentiation.

Evaluation of Air Pollution Control Devices for Asphalt Pavement Recycling Operations, HR-188, Progress Report, 1977

This report discusses the asphalt pavement recycling project designated Project HR-188 in Kossuth County, Iowa. Specific objectives were: (a) to determine the effectiveness of drum mixing plant modifications designed to control air pollution within limits specified by the Iowa Department of Environmental Quality; (b) to assess the impact of varying the proportions of recycled and virgin aggregates, (c) to assess the impact of varying the production rate of the plant, and (d) to assess the impact of varying the mixing temperature. The discussion includes information on the proposed use of research funds, project location and description, the project planning conference, plan development, bid letting, asphalt plant configuration, actual plant operation, why this method is successful, probable process limitations, pollution results, recycled pavement test results, and the cost of virgin vs. recycled asphalt pavements.

Dust Control Using an Asphalt Emulsion, HR-196, 1977

In an effort to control fugitive dust on a gravel surfaced roadway in Boone County, a cationic asphalt emulsion was blended with warm water and applied with an asphalt distributor. The test included various application procedures. After visual observations, it was concluded that this procedure utilizing a dilute asphalt emulsion was not an effective method of dust control.

Improved Asphalt Surfaces and Asphalt Resurfacing Performance through Crack Maintenance, HR-213, 1987

In 1980, a Vanguard High Pressure Water Blaster capable of providing 10 gallons of water per minute at 2000 psi was purchased to evaluate water blasting as a crack cleaning method prior to crack filling on asphalt concrete pavements. Afer some iniital trials demonstrated its effectiveness of removing dirt, debris and vegetation, it was included in joint and crack maintenance research on Iowa 7 in Webster County. The objective of the research was to evaluate six crack preparation methods and seven "sealant" materials. The cleaning and sealing was performed in the spring of 1983. Visual evaluations of the performance were made in the fall of 1983 and spring of 1985. Compressed air and/or high pressure water did not adequately prepare cracks less than 3/8 inch wide. Routing or sawing was necessary to provide a sealant reservoir. The water blaster was more effective than compressed air in removing dirt, debris and vegetation but this did not yield significant improvement in sealant adhesion or longevity. Periodic crack filling is necessary on ACC surfaces throughout the remaining life of the pavement.

Asphalt Emulsion Bound Macadam in Dubuque County, Iowa, HR-216, Construction Report, 1981

This construction report describes a project investigating macadam base construction of roads. The objectives of this project were to: 1. identify a cost effective asphalt emulsion bound macadam typical cross section; 2. obtain useful data comparing seven typical cross sections; 3. determine the effectiveness of engineering fabric placed under macadam roadbeds; and 4. evaluate the use of emulsions in surface seal coats.

Evaluation of Recycled Rubber in Asphalt Concrete - Plymouth County, HR-330A, 1992

The Iowa Department of Transportation is evaluating the use of ground recycled crumb rubber from discarded tires in asphalt rubber cement. There were four projects completed during 1991 and another one constructed in 1992. This project is located on IA 140 north of Kingsley in Plymouth County. The project contains one section with reacted asphalt rubber cement (ARC) used in both binder and surface courses, one with reacted ARC used in the surface course and a conventional binder course, and a conventional mix control section. The reacted rubber binder course was placed on October 17, 1991 and the reacted rubber surface course was placed on October 17, 18, and 19. Inclement weather caused a slight delay in placing or constructing the surface. There was a minor problem with shoving and cracking of the binder course. The construction went well otherwise. Information included in this report consists of test results, construction reports, and cost comparisons.

Evaluation of Recycled Rubber in Asphalt Concrete - Dubuque County, HR-330C, 1992

The Iowa Department of Transportation is evaluating the use of discarded tires in asphalt rubber cement. There have been five projects completed in Iowa. This project is located on US 151 north of Cascade to US 61 in Dubuque. One section consists of an asphalt rubber cement surface and a conventional binder and two sections contain both asphalt rubber cement surface and binder. The control section of conventional asphalt was completed this spring. Information included in this report consists of test results, construction reports, and cost comparisons.

Evaluation of Recycled Rubber in Asphalt Concrete - Black Hawk County, HR-330D, 1992

The disposal of discarded tires has become a major problem. Different methods of recycling have been researched. Currently, Iowa is researching the use of ground recycled crumb rubber from discarded tires in asphalt rubber cement. Six projects have been completed in Iowa using asphalt rubber cement. This project is located on IA 947 (University Avenue) in Cedar Falls/Waterloo. The project contains one section with asphalt rubber cement used in both the binder and surface courses and one section using asphalt rubber cement in the surface course with a conventional binder. There are two control sections where conventional asphalt pavement was placed.

Field Evaluation of Engineering Fabrics for Asphalt Concrete Resurfacing - Audubon County, HR-360, Construction Report, 1994

An asphalt concrete (ACC) overlay is most often the rehabilitative effort used to maintain the serviceability of either an ACC or PCC pavement. The major problem in durability of this ACC overlay comes from reflective cracking. These cracks usually open, allowing water to enter the unsealed crack and strip the ACC in the overlay. The stripping of the ACC allows accelerated deterioration at the crack. Two engineering fabrics were evaluated in this project in order to determine their effectiveness in reducing reflective cracking. These two materials are: PavePrep, Contech Construction Products, Inc., and Pro-Guard, Phillips Fiber Corporation. A 4.2 km (2.6 mi) roadway in Audubon County was selected for the research project. The roadway was divided into eight test sections. Four of the test sections are conventional resurfacing. The other four sections are split between the two engineering fabrics (two Pro-Guard and two PavePrep). A 75 mm (3 in.) thick overlay was placed over the entire project.

Polymerized Asphalt Cement, HR-533, 1989

In 1986, a 0.34 mile experimental section of polymerized asphalt cement (PAC30) concrete was placed in the westbound driving lane of Interstate 80 in western Iowa. It was used in a 2" asphalt concrete inlay using 20% recycled asphalt pavement. The virgin aggregate included 41% crushed gravel, 25% crushed quartzite and 14% natural sand. The evaluation of the project was severely limited when a 1987 reconstruction project extended into the experimental section leaving only 395 feet. Rut depths under a 4-foot gage were taken for a period of two years. No significant rutting occurred in the experimental polymerized section. The frequency of transverse cracking in the polymerized AC section was the same as that of the comparative AC-20 section. The asphalt paving mixture made with polymerized AC cost 120% of the cost of the conventional mix.

Evaluation of Asphalt Stabilizing Additives, HR-542, 1994

Efforts are constantly being put forth by researchers, highway related industries and product suppliers to improve the life and performance of asphalt pavements. As a result of those efforts, a variety of asphalt modifiers have been developed and evaluated in experimental sections over the years. Evaluations of the polymer asphalt modifiers have been done and results were usually compared with conventional sections within each respective project. The research presented in this report is also a comparison of asphalt modifiers with each other as well as a comparison of a modifier with its respective conventional section, when they exist. Several of the modifiers showed some improvements in performance while others did not.