Rapid IL-4 production by Leishmania homolog of mammalian RACK1-reactive CD4(+) T cells in resistant mice treated once with anti-IL-12 or -IFN-gamma antibodies at the onset of infection with Leishmania major instructs Th2 cell development, resulting in nonhealing lesions.

Rapid production of IL-4 by Leishmania homolog of mammalian RACK1 (LACK)-reactive CD4(+) T cells expressing the V beta 4-V alpha 8 TCR chains has been shown to drive aberrant Th2 cell development and susceptibility to Leishmania major in BALB/c mice. In contrast, mice from resistant strains fail to express this early IL-4 response. However, administration of either anti-IL-12 or -IFN-gamma at the initiation of infection allows the expression of this early IL-4 response in resistant mice. In this work we show that Leishmania homolog of mammalian RACK1-reactive CD4(+) T cells also expressing the V beta 4-V alpha 8 TCR chains are the source of the early IL-4 response to L. major in resistant mice given anti-IL-12 or -IFN-gamma Abs only at the onset of infection. Strikingly, these cells were found to be required for the reversal of the natural resistance of C57BL/6 mice following a single administration of anti-IL-12 or -IFN-gamma Abs. Together these results suggest that a deficiency in mechanisms capable of down-regulating the early IL-4 response to L. major contributes to the exquisite susceptibility of BALB/c mice to L. major.

Different antigen-presenting cells differ in their capacity to induce lymphokine production and proliferation of an apo-cytochrome c-specific T cell clone.

The activation of an apo-cytochrome c-specific T cell clone was found to differ, depending on the antigen-presenting cell population. Whereas total syngeneic spleen cells and bone marrow macrophages could be shown to trigger proliferation, IL 2, and MAF production by the T cell clone, a B cell lymphoma only induced MAF secretion. Further studies demonstrated that this effect was not due to a different antigen processing by the B lymphoma or to limiting amounts of Ia and antigen molecules on the B lymphoma cell surface. The dissociation of induction of MAF production from IL-2 production/proliferation found with the different antigen-presenting cells indicates strongly that molecules other than Ia and antigen may be required for the complete functional activation of antigen-specific T cell clones.

T-cell hybridoma specific for a cytochrome c peptide: specific antigen binding and interleukin 2 production.

T-cell hybridomas were obtained after fusion of BW 5147 thymoma and long-term cultured T cells specific for cytochrome c peptide 66-80 derivatized with a 2,4-dinitroaminophenyl (DNAP) group. The resulting hybridomas were selected for their capacity to specifically bind to soluble radiolabeled peptide antigen. One T-cell hybrid was positive for antigen binding. This hybrid T cell exhibits surface phenotypic markers of the parent antigen-specific T cells. The binding could be inhibited either by an excess of unlabeled homologous antigen or by cytochrome c peptide 11-25 derivatized with a 2-nitrophenylsulfenyl group. Several other peptide antigens tested failed to inhibit binding of the radioactive peptide. This suggests that a specific amino acid sequence, modified by a DNAP group, is the antigenic structure recognized by the putative T-cell receptor. In addition, direct interaction of DNAP-66-80 peptide with the hybridoma cell line induced production of the T-cell growth factor interleukin 2. Furthermore, supernatants derived from syngeneic macrophages pulsed with the relevant peptide also induced the antigen-specific hybridoma to produce interleukin 2.

Behavior of Asphalts in the Production of Asphaltic Concrete, HR-107, 1965

Project 540-S of the Iowa Engineering Experiment Station (Project HR-107, Iowa Highway Research Board) was started in June, 1964. During the year ten 2-gallon samples of asphalt cement and ten 100-lb samples of asphaltic concrete were studied by the personnel of the Bituminous Research Laboratory, Iowa State University. The samples were from tanks and mixers of asphalt plants at various Iowa State Highway Commission paving jobs. The laboratory's research was in two phases: 1. To ascertain if properties of asphalt cement changed during mixing operations. 2. To determine whether one or more of the several tests of asphalt cements were enough to indicate behavior of the heated asphalt cements. If the reliability of one or more tests could be proved, the behavior of asphalts would be more simply and rapidly predicted.

Production of Asphalt Rubber Concrete by the Dry Process, HR-1062, 1994

Disposal of used tires has been a problem throughout the United States. The 1991 Intermodal Surface Transportation Efficiency Act (ISTEA) requires the use of recycled rubber in asphalt concrete starting in FY94. A moratorium has delayed this requirement until FY95. The Iowa DOT has researched six projects using crumb rubber modifier in asphalt concrete using the wet process. This process involves using a blender-reactor to blend the asphalt cement and crumb rubber. Using the wet process the asphalt cement has to reach a hotter temperature, than is normally required, for reaction to occur. The wet process is also much more expensive than conventional asphalt. This research deals with using a dry process to incorporate crumb rubber into the asphalt concrete mix. The project was constructed by Western Engineering of Harlan, Iowa, on IA 37 between Earling, Iowa and US 59. It was completed in September 1993. Western Engineering used a double drum mixer to produce the crumb rubber modified asphalt concrete by the dry process. The production and construction went well with minor difficulty and the dry process is a less expensive procedure for producing crumb rubber modified asphalt concrete.

Rapid, simple and high yield production of recombinant proteins in mammalian cells using a versatile episomal system.

Many research projects in life sciences require purified biologically active recombinant protein. In addition, different formats of a given protein may be needed at different steps of experimental studies. Thus, the number of protein variants to be expressed and purified in short periods of time can expand very quickly. We have therefore developed a rapid and flexible expression system based on described episomal vector replication to generate semi-stable cell pools that secrete recombinant proteins. We cultured these pools in serum-containing medium to avoid time-consuming adaptation of cells to serum-free conditions, maintain cell viability and reuse the cultures for multiple rounds of protein production. As such, an efficient single step affinity process to purify recombinant proteins from serum-containing medium was optimized. Furthermore, a series of multi-cistronic vectors were designed to enable simultaneous expression of proteins and their biotinylation in vivo as well as fast selection of protein-expressing cell pools. Combining these improved procedures and innovative steps, exemplified with seven cytokines and cytokine receptors, we were able to produce biologically active recombinant endotoxin free protein at the milligram scale in 4-6weeks from molecular cloning to protein purification.

IL-1β Suppresses Innate IL-25 and IL-33 Production and Maintains Helminth Chronicity.

Approximately 2 billion people currently suffer from intestinal helminth infections, which are typically chronic in nature and result in growth retardation, vitamin A deficiency, anemia and poor cognitive function. Such chronicity results from co-evolution between helminths and their mammalian hosts; however, the molecular mechanisms by which these organisms avert immune rejection are not clear. We have found that the natural murine helminth, Heligmosomoides polygyrus bakeri (Hp) elicits the secretion of IL-1β in vivo and in vitro and that this cytokine is critical for shaping a mucosal environment suited to helminth chronicity. Indeed in mice deficient for IL-1β (IL-1β(-/-)), or treated with the soluble IL-1βR antagonist, Anakinra, helminth infection results in enhanced type 2 immunity and accelerated parasite expulsion. IL-1β acts to decrease production of IL-25 and IL-33 at early time points following infection and parasite rejection was determined to require IL-25. Taken together, these data indicate that Hp promotes the release of host-derived IL-1β that suppresses the release of innate cytokines, resulting in suboptimal type 2 immunity and allowing pathogen chronicity.

Behavior of Asphalts During Production of Asphaltic Concrete Mixes, HR-107, 1965

When mixing asphalt in thin film and at high temperatures, as in the production of asphalt concrete, it has been shown that asphalt will harden due essentially to two factors: (1) losses of volatiles and (2) oxidation. The degree of hardening as expressed by percent loss in penetration varied from as low as 7% to about 57% depending on mixing temperatures, aggregate types, gradation, asphalt content, penetration and other characteristics of asphalts used. Methods used to predict hardening during mixing include loss on heat and thin film oven tests, with the latter showing better correlation with the field findings. However, information on other physical and chemical changes that may occur as a result of mixing in the production of hot-mix asphaltic concrete is limited, The purpose of this research project was to ascertain the changes of asphalt cement properties, both physical and chemical, during mixing operation and to determine whether one or more of the several tests of asphalt cements were critical enough to indicate these changes.

Biotechnological production of taxanes: a molecular approach

Plant cell cultures constitute a promise for the production of a high number of phytochemicals, although the majority ofbioprocesses that have been developed so far have not resultedcommercially successful. An overview indicates that most of theresearch carried out until now is of the empirical type. For this reason,there is a need for a rational approach to the molecular and cellularbasis of metabolic pathways and their regulation in order to stimulatefuture advances.The empirical investigations are based on the optimization of theculture system, exclusively considering input factors such as theselection of cellular lines, type and parameters of culture, bioreactordesign and elicitor addition, and output factors such as cellular growth,the uptake system of nutrients, production and yield. In a rationalapproach towards the elucidation of taxol and related taxaneproduction, our group has studied the relationship between the taxaneprofile and production and the expression of genes codifying forenzymes that participate in early, intermediate and late steps of theirbiosynthesis in elicited Taxus spp cell cultures. Our results show that elicitors induce a dramatic reprogramming of gene expression in Taxus cell cultures, whichlikely accounts for the enhanced production of taxol and related taxanes and we have alsodetermined some genes that control the main flux limiting steps. The application ofmetabolic engineering techniques for the production of taxol and taxanes of interest is also discussed.

Effect of intercropping wheat with forage legumes on wheat production and ground cover

The use of winter legumes in southern Brazil is hindered by the slow growth of these species during establishment exposing soil surface to erosion. Introduction of these species along with spring wheat (Triticum aestivum L.) was studied as a means of increasing ground cover during their initial establishment period, without reducing wheat grain yield. Two experiments were conducted in nearby areas, one in each year. Birdsfoot trefoil (Lotus corniculatus L.), red clover (Trifolium pratense L.) cultivar Quiñequelli, white clover (T. repens L.), and arrowleaf clover (T. vesiculosum Savi) did not reduce cereal yield in either year. Wheat yield was reduced by intercropped red clover cultivar Kenland and by subclover (T. subterraneum L.) in the first year. No grain yield differences due to intercropping with any legume were detected in the second year, when rainfall was below normal. Intercropping with wheat showed to be a practical alternative to enhance ground cover at establishing forage legumes.

Effects of group size and space allocation on physiological, behavioural and production-related welfare parameters in farmed silver fox cubs

Selostus: Ryhmäkoon ja käytössä olevan tilan vaikutus tarhattujen hopeakettupentujen hyvinvointiin

Can protoplast production from in vitro cultured shoots of Tanacetum vary during the season?

Selostus: Vaikuttaako vuodenaika Tanacetum-lajien in vitro kasvuun ja eristettävien protoplastien määrään?

Non-viral gene therapy for GDNF production in RCS rat: the crucial role of the plasmid dose.

Glial cell line-derived neurotrophic factor (GDNF) is one of the candidate molecules among neurotrophic factors proposed for a potential treatment of retinitis pigmentosa (RP). It must be administered repeatedly or through sustained releasing systems to exert prolonged neuroprotective effects. In the dystrophic Royal College of Surgeon's (RCS) rat model of RP, we found that endogenous GDNF levels dropped during retinal degeneration time course, opening a therapeutic window for GDNF supplementation. We showed that after a single electrotransfer of 30 μg of GDNF-encoding plasmid in the rat ciliary muscle, GDNF was produced for at least 7 months. Morphometric, electroretinographic and optokinetic analyses highlighted that this continuous release of GDNF delayed photoreceptors (PRs) as well as retinal functions loss until at least 70 days of age in RCS rats. Unexpectedly, increasing the GDNF secretion level accelerated PR degeneration and the loss of electrophysiological responses. This is the first report: (i) demonstrating the efficacy of GDNF delivery through non-viral gene therapy in RP; (ii) establishing the efficacy of intravitreal administration of GDNF in RP associated with a mutation in the retinal pigment epithelium; and (iii) warning against potential toxic effects of GDNF within the eye/retina.