A monoclonal antibody to equine interleukin-4

Interleukin-4 (IL-4) is secreted by T helper type 2 cells, mast cells, basophils and eosinophils. Detection of IL-4 can contribute the evaluation of cellular immune responses during infectious diseases, immunological disorders or vaccination. We used recombinant equine IL-4 to generate a monoclonal antibody (mAb) to equine IL-4. The mAb detected recombinant IL-4 in mammalian cells transfected with different plasmids containing IL-4 cDNA. After mitogen stimulation of equine peripheral blood mononuclear cells, an intracellular protein was recognized by the new mAb in 1–2% of lymphocytes using flow cytometric analysis. In the presence of the secretion blocker Brefeldin A, the protein accumulated and was detected in 4–8% of lymphocytes stimulated with phorbol 12-myristate 13-acetate and ionomycin. Double staining with the new mAb and T-cell or B-cell markers identified a subpopulation of CD4⁺ T-cells expressing the protein recognized by the mAb. In addition, the protein was detectable in cell culture supernatants of mitogen stimulated cells by ELISA when using the new mAb for coating of the plates and a polyclonal antiserum to equine IL-4 for detection. In conclusion, the new mAb detects equine IL-4 and can be used for intracellular staining and ELISA to measure this important cytokine.     

The competitive advantage of sanctioning institutions

Understanding the fundamental patterns and determinants of human cooperation and the maintenance of social order in human societies is a challenge across disciplines. The existing empirical evidence for the higher levels of cooperation when altruistic punishment is present versus when it is absent systematically ignores the institutional competition inherent in human societies. Whether punishment would be deliberately adopted and would similarly enhance cooperation when directly competing with nonpunishment institutions is highly controversial in light of recent findings on the detrimental effects of punishment. We show experimentally that a sanctioning institution is the undisputed winner in a competition with a sanction-free institution. Despite initial aversion, the entire population migrates successively to the sanctioning institution and strongly cooperates, whereas the sanction-free society becomes fully depopulated. The findings demonstrate the competitive advantage of sanctioning institutions and exemplify the emergence and manifestation of social order driven by institutional selection.     

Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis

Background: Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. Objectives: The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Methods: Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–Whitney U test. Results: Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α₁‐globulin (1.9–3.1 g/L), α₂‐globulin (5.3–8.7 g/L), β₁‐globulin (2.8–7.3g/L), β₂‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65). Conclusion: Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.     

Diversity,spatial variation,and temporal dynamics of virulences in the German leaf rust (<Emphasis Type="Italic">Puccinia recondita</Emphasis> f. sp. <Emphasis Type="Italic">secalis</Emphasis>) population in winter rye

A large collection of German rye leaf rust isolates was analysed to characterize the diversity, spatial variation and temporal dynamics of virulences. Virulence-avirulence phenotypes (=pathotypes) were determined on 23 host differentials. We found 93 pathotypes among 177 single-uredinial isolates in 2000, 201 pathotypes among 437 isolates in 2001, and 125 pathotypes among 213 isolates in 2002. In total, the 827 analyzed isolates represented 317 pathotypes. Frequency of virulences on the individual differentials varied from 2% to 97%. Eight of the differentials showed a high resistance level with virulence frequencies <10%. Virulence complexity of the isolates ranged from 3 to 21 with a mean of nine. The percentages of highly virulent isolates (>14 virulences) increased from 4 to 15% during the sampling period. A high level of virulence diversity was observed within and between individual sampling sites with Simpson indices around 0.9. Evenness indices ranged from 0.88 to 0.92. Four of the five most frequent pathotypes were found in each year but their frequency never exceeded 10%. Isolates with unusual virulence combinations could be clearly separated by principal component analysis. Location-specific pathotype frequencies were revealed in each year, but the frequency patterns varied across years. On four fields a considerable increase of highly virulent pathotypes occurred within 6 weeks during the epidemic. The high diversity of pathotypes as well as the fast accumulation of highly virulent pathotypes favour the adaptation of the pathogen to race-specific host resistances. More durable resistance might be achievable by combining new effective race-specific resistances with adult-plant and/or race-non-specific quantitative resistances.     

Model‐based evaluation of agri‐environmental measures in the Federal State of Brandenburg (Germany) concerning N pollution of groundwater and surface water

GIS‐based modeling of soil‐crop interactions and hydrological processes is a valuable instrument to assess land‐use effects on N pollution of water resources from the agricultural sector. A case study is presented using spatial information on soils, climatic zones, land use, and distribution of agri‐environmental measures within the federal State of Brandenburg (Germany) to assess the reduction effect of EU‐funded measures on N pollution of water resources. In a first step, the area was classified concerning the risk for groundwater and surface‐water pollution. For this, spatially distributed model calculations of the soil‐solution exchange frequency were intersected to a vulnerability map for groundwater derived from geological data and zones of different transit times from the root zone into surface waters. In a second step, model calculations of water and N dynamics in the soil‐crop system for different crop and management systems were performed to calculate nitrate leaching from the root zone and to estimate the effect of present agri‐environmental measures to reduce N pollution on groundwater and surface waters. The results indicated that 75% of the agri‐environmental measures were placed in areas with low impact on groundwater and surface waters. Therefore, the effectiveness of the agri‐environmental measures concerning water‐protection aims was moderate.     

Intact carbohydrate structures as part of the melanoidin skeleton

Model melanoidins from monomeric, oligomeric, and polymeric carbohydrates, and amino acids formed under aqueous as well as water-free reaction conditions, were submitted to acidic catalyzed hydrolysis. Their degradation products were detected qualitatively and quantitatively by HPTLC and HPLC-DAD. A considerable amount of monomer carbohydrates from hydrolysis of model melanoidins formed under water-free reaction conditions was detected. It can be seen clearly that the amount of carbohydrates released increased with increasing degree of polymerization of the carbohydrates used as starting material. In comparison, the hydrolysis of melanoidins formed in aqueous condition resulted in only a small glucose release. It seems that in the Maillard reaction under water-free conditions, a significant amount of di- and oligomer carbohydrates were incorporated into the melanoidin skeleton as complete oligomer with intact glycosidic bond, forming side chains at the melanoidin skeleton. Additional side chains could be formed by transglycosylation reactions. With increasing water content, hydrothermolytic as well as retro-aldol reactions of the starting carbonyl components became significant, and therefore the possibility of forming side chains decreased. The results are consistent with the postulated melanoidin structure being built up mainly from sugar degradation products, probably branched via amino compounds.     

Evaluation of immune responses following infection of ponies with an EHV-1 ORF1/2 deletion mutant

ABSTRACT: Equine herpesvirus-1 (EHV-1) infection remains a significant problem despite the widespread use of vaccines. The inability to generate a protective immune response to EHV-1 vaccination or infection is thought to be due to immunomodulatory properties of the virus, and the ORF1 and ORF2 gene products have been hypothesized as potential candidates with immunoregulatory properties. A pony infection study was performed to define immune responses to EHV-1, and to determine if an EHV-1 ORF1/2 deletion mutant (ΔORF1/2) would have different disease and immunoregulatory effects compared to wild type EHV-1 (WT). Infection with either virus led to cytokine responses that coincided with the course of clinical disease, particularly the biphasic pyrexia, which correlates with respiratory disease and viremia, respectively. Similarly, both viruses caused suppression of proliferative T-cell responses on day 7 post infection (pi). The ΔORF1/ORF2 virus caused significantly shorter primary pyrexia and significantly reduced nasal shedding, and an attenuated decrease in PBMC IL-8 as well as increased Tbet responses compared to WT-infected ponies. In conclusion, our findings are (i) that infection of ponies with EHV-1 leads to modulation of immune responses, which are correlated with disease pathogenesis, and (ii) that the ORF1/2 genes are of importance for disease outcome and modulation of cytokine responses.     

Intraocular nematodiasis in a llama (Lama glama)

This report describes a unique case of presumed migration of Parelaphastrongylus tenuis through the spinal cord into the eye of a llama where it survived and matured within the ocular environment. Blindness of the eye was most likely attributable to migration of the parasite through the central nervous tissue.