Time-dependent alterations in gene expression of interleukin-8 in the bronchial epithelium of horses with recurrent airway obstruction

OBJECTIVE: To evaluate time-dependent alterations in gene expression of chemokines in bronchial epithelium of recurrent airway obstruction (RAO)-affected horses and whether alterations resulted from increases in gene expression of interleukin (IL)-17 in cells isolated from bronchoalveolar lavage fluid (BALF). ANIMALS: 8 RAO-susceptible horses and 9 control horses. PROCEDURE: In 2 experiments, both groups of horses were evaluated after being maintained on pasture and after being stabled and fed dusty hay for 1, 14, 35, and 49 days (experiment 1) or 14 and 28 days (experiment 2). In experiment 1, gene expression of IL-8, chemokine (C-X-C motif) ligand 1 (CXCL1), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and Toll-like receptor 4 (TLR4) in epithelium and IL-8, IL-17, and TLR4 in BALF cells was measured. In experiment 2, bronchial biopsy specimens were evaluated for IL-8 immunoreactivity. RESULTS: In RAO-susceptible horses after 14 days of challenge exposure, there was a 3- and 10-fold increase in gene expression of IL-8 for epithelial and BALF cells and an increase in IL-8 immunoreactivity in epithelial cells. Challenge exposure failed to alter gene expression of CXCL1, GM-CSF, G-CSF, and TLR4 in epithelial cells of any horses at any time point. During challenge exposure, gene expression of BALF cell IL-17 was downregulated in control horses (day 1) and upregulated in RAO-affected horses (day 35). CONCLUSIONS AND CLINICAL RELEVANCE: Epithelial-derived IL-8 may promote airway neutrophilia, but the inciting stimulus is unlikely to be IL-17 because upregulation of this gene is subsequent to that of IL-8 in epithelial cells.     

CO<Subscript>2</Subscript>-assimilation and chlorophyll fluorescence as indirect selection criteria for host tolerance against <Emphasis Type="Italic">Striga</Emphasis>

Striga hermonthica (Del.) Benth. is a parasitic weed on tropical cereals causing serious yield losses in Africa. The use of host crop varieties with improved resistance and tolerance against this parasite is a key component of an integrated control strategy. Breeding for tolerance is however seriously hampered by the absence of reliable and yet practical selection measures. The observation that the photosynthetic rate of tolerant genotypes is less sensitive to Striga infection was used as a starting point to search for suitable selection measures. In a greenhouse pot experiment the effect of Striga infection on the photosynthesis of four sorghum (Sorghum bicolor [L.] Moench) genotypes, differing in Striga tolerance level, was measured at three moments in time (26, 48 and 75 days after sowing). Genotypes were CK60-B, E36-1, Framida and Tiémarifing. Measurements involved CO₂-assimilation (A) and three chlorophyll fluorescence characteristics (electron transport rate through photosystem II [ETR], photochemical [Pq] and non-photochemical quenching [NPq]). Striga infection negatively affected A, ETR and Pq. Based on A and Pq, genotypes with superior levels of tolerance (Tiémarifing) could be discriminated from genotypes with superior level of resistance (Framida). Both A and Pq showed high heritabilities and consequently clear and predictable differences between genotypes. Using discriminative ability, heritability and cost effectiveness as main criteria, photochemical quenching (Pq) was concluded to possess the highest potential to serve as indirect selection measure for host plant tolerance to Striga. Screening should preferably be conducted at relatively high Striga infestation levels, between Striga emergence and host plant flowering.     

Effect of grazing intensity and soil characteristics on soil organic carbon and nitrogen stocks in a temperate long-term grassland

The effects of different grazing pressures (GPs) on soil properties are not sufficiently understood. The objectives were to analyse the effects of three different extensive GPs on stocks of soil organic C and total N, soil microbial biomass C, basal respiration and mineral N in three different soil depths of a long-term pasture in Central Germany (FORBIOBEN field trial). No significant (p ≤ 0.05) effects of GP on weighted stocks of soil organic C, total N, soil microbial biomass C, mineral N and basal respiration rate were observed, suggesting that the C and N cycles are coupled in the three grazing treatments. Oxalate soluble Fe contents explained a marked part of the variation of soil organic C (multiple linear regression: R² = 0.64) and total N contents (R² = 0.64) in the soils, whereas almost all of the variability of soil microbial biomass C contents and basal respiration was explained by soil organic C contents. Overall, variabilities of soil organic C and N contents were largely explained by oxalate soluble Fe contents, whereas grazing intensity did not affect the C and N dynamics.     

Diversity of bacterial laccase-like multicopper oxidase genes in forest and grassland Cambisol soil samples

Laccases- or laccase-like multicopper oxidases (LMCO) catalyze the oxidation of various substrates, such as phenols, diamines and metals, coupled with the reduction of molecular oxygen to water. Compared to studies on function and diversity of LMCO in plants and fungi, little is known about this enzyme type in bacteria and especially on their possible implication in degradation of organic matter in soils. This study presents a molecular investigation of the diversity and distribution of bacterial LMCO genes among three upper horizons of a forest Cambisol and in a grassland Cambisol. Some culture strains of soil bacteria were also analyzed at the molecular level and for their capability to oxidize naturally occurring 2,6-dimethoxyphenol, a LMCO substrate. A high LMCO gene diversity was found in the Cambisol soil samples with 16 distinct sequence type clades, of which approximately one half was not matching with any reference sequence of known bacteria. The highest richness of bacterial LMCO genes was observed in the organic horizon of the forest soil, which is concomitant with a previous analysis of the diversity of fungal laccase genes and corresponding soil laccase activity. Some clusters of sequence types showed a specific distribution in one of the soils or in horizons, while others appeared more ubiquist. Multiple bacterial LMCO genes were described in Agromyces salentinus and Sinorhizobium morelense, what so far was only known from fungi.     

Feasibility of using retinol-binding protein from capillary blood specimens to estimate serum retinol concentrations and the prevalence of vitamin A deficiency in low-resource settings

Vitamin A deficiency (VAD) is a significant public health problem in many countries. While cost-effective interventions are available to control VAD, reliable information is needed to the track progress of control programmes. However, assessment of VAD is uncommon because current approaches are expensive and not feasible in low-resource settings. The present study explores the utility of retinol-binding protein (RBP), analysed by enzyme-linked immunosorbent assay from capillary blood, as an alternative measure of serum retinol concentrations in populations. The study collected matched panels of venous and capillary blood from pre-school children in Chiang Mai, Thailand. Of a total sample of 195 children, there were no differences between RBP from venous blood, RBP from capillary blood or retinol from capillary blood relative to retinol from venous blood. Receiver-operating characteristic curve analysis suggested a cut-off of RBP < 0.825 micromol l(-1) had optimal screening proficiency relative to retinol <0.70 micromol l(-1). For the purpose of population assessment, all three parameters performed well in screening for VAD relative to retinol from venous blood. There were no differences in the estimates of VAD between children stratified by inflammation status. Lower RBP concentrations were found in children in the early convalescent stage of infection than in children with no infection or in the late convalescent stage. This study provided evidence of the biological comparability between retinol and RBP estimated from venous blood and capillary blood. This is a critical observation as it provides empirical evidence that RBP from capillary blood is a surrogate measure of serum retinol concentrations.     

Genotype × environment interaction and yield stability in barley (Hordeum vulgare L.) genotypes in the central highland of Ethiopia

Journal of Crop Science and Biotechnology - Barley is one of the most important cereal crops cultivated over a wider environment in the diverse agro-ecologies in Ethiopia. Study on genotype by...     

Reduced incidence of insect-bite hypersensitivity in Icelandic horses is associated with a down-regulation of interleukin-4 by interleukin-10 and transforming growth factor-beta1

Insect bite hypersensitivity (IBH) is an allergic dermatitis of horses caused by IgE-mediated reactions to bites of insects of the genus Culicoides. IBH does not occur in Iceland due to the absence of Culicoides. However, Icelandic horses exported to mainland Europe as adults (1st generation) have a > or =50% incidence of developing IBH. In contrast, their progeny (2nd generation) has a <10% incidence of IBH. Here we show that peripheral blood mononuclear cells (PBMC) from Icelandic horses born in mainland Europe and belonging either to the IBH or healthy subgroup produce less interleukin (IL)-4 after polyclonal or allergen-specific stimulation when compared with counterparts from horses born in Iceland. We examined a role of IL-10 and transforming growth factor (TGF)-beta1 in down-regulation of IL-4 in healthy 2nd generation Icelandic horses. Supernatants of PBMC from 2nd generation healthy horses down-regulated the proportion of IL-4-producing cells and IL-4 production in stimulated cultures of PBMC from 1st generation IBH. This inhibition was mimicked by a combination of IL-10 and TGF-beta1 but not by the single cytokines. Cultures of stimulated PBMC of healthy 2nd generation horses produced a low level of IL-4, but IL-4 production was increased by anti-equine IL-10 and anti-human TGF-beta1. This shows for the first time that in horses, IL-10 and TGF-beta1 combined regulate IL-4 production in vitro. It is suggested that in this naturally occurring IgE-mediated allergy, IL-10 and TGF-beta1 have a role in the down-regulation of IL-4-induced allergen-specific Th2 cells, thereby reducing the incidence of IBH.     

A histamine release assay to identify sensitization to Culicoides allergens in horses with skin hypersensitivity

Skin hypersensitivity is an allergic disease induced in horses by allergens of Culicoides midges. The condition is typically diagnosed by clinical signs and in some horses in combination with allergy testing such as intradermal skin testing or serological allergen-specific IgE determination. Here, we describe an alternative method for allergy testing: a histamine release assay (HRA) that combines the functional aspects of skin testing with the convenience of submitting a blood sample. The assay is based on the principle that crosslinking of allergen-specific IgE bound via high-affinity IgE receptors to the surfaces of mast cells and basophils induces the release of inflammatory mediators. One of these mediators is histamine. The histamine was then detected by a colorimetric enzyme-linked immunosorbent assay. The histamine assay was used to test 33 horses with skin hypersensitivity and 20 clinically healthy control animals for histamine release from their peripheral blood basophils after stimulation with Culicoides allergen extract or monoclonal anti-IgE antibody. An increased histamine release was observed in the horses with skin hypersensitivity compared to the control group after allergen-specific stimulation with Culicoides extract (p=0.023). In contrast, stimulation with anti-IgE induced similar amounts of released histamine in both groups (p=0.46). For further evaluation of the HRA, we prepared a receiver operating-characteristic (ROC) curve and performed a likelihood-ratio analysis for assay interpretation. Our results suggested that the assay is a valuable diagnostic tool to identify sensitization to Culicoides allergens in horses. Because some of the clinically healthy horses also showed sensitization to Culicoides extract, the assay cannot be used to distinguish allergic from non-allergic animals. The observation that sensitization is sometimes detectable in non-affected animals suggested that clinically healthy horses use immune mechanisms to control the reaction to Culicoides allergens that are different or absent in allergic horses.     

Characterization of Streptococcus equi subsp. ruminatorum isolated from spotted hyenas (Crocuta crocuta) and plains zebras (Equus burchelli), and identification of a M-like protein (SrM) encoding gene

Thirteen strains of Streptococcus equi subsp. ruminatorum from free-ranging spotted hyenas (Crocuta crocuta) and plains zebras (Equus burchelli) in Tanzania were characterized by biochemical and molecular-biological methods. Although the colony appearance of the S.e. ruminatorum wildlife strains differed from that of the S.e. ruminatorum type strain CECT 5772(T), all biochemical reactions of the wildlife strains were similar to those of the type strain. In addition, all wildlife strains produced hyaluronidase and were capable of hydrolysing arginine, three strains (23%) synthesized acetoin, but only eight strains (62%) produced acid from ribose. rep-PCR indicated that different clones of S.e. ruminatorum were distributed among the hyena and zebra populations in the study area. Identical rep-PCR patterns in hyena and zebra strains suggest that a direct transmission of S.e. ruminatorum between these species may occur. The presence of a M-like protein (SrM) gene was demonstrated in all S.e. ruminatorum strains including the type strain. Sequencing of the M-like protein gene revealed a hypervariable region within the deduced amino acid sequence. Most of the strains clustered with previously described strains based on the hypervariable region of the S.e. zooepidemicus SzP protein. Sequencing also demonstrated that identical SrM protein sequences were shared among S.e. ruminatorum strains from different host species.