Co-incubation with IL-18 potentiates antigen-specific IFN-γ response in a whole-blood stimulation assay for measurement of cell-mediated immune responses in pigs experimentally infected with Lawsonia intracellularis

The whole-blood interferon-gamma (IFN-γ) assay is a quantitative in vitro assay for a direct read-out of Ag-specific cell-mediated immune (CMI) responses to infectious diseases. The IFN-γ assay is robust in severe intracellular infections like Brucella or mycobacteria, but more difficult to evaluate for less severe or immunocompromising infections. Here we investigated the performance of the assay when recombinant co-stimulatory cytokines IL-12 and/or IL-18 were added along with Ag or PBS to cultures of whole-blood from pigs infected with Lawsonia intracellularis. In pigs recovering from a natural infection, addition of rIL-12 or rIL-18 alone increased the Ag-specific IFN-γ release while addition of both cytokines resulted in increased IFN-γ release also in PBS cultures. In analyses after experimental infections with L. intracellularis, significant increased levels of Ag-specific IFN-γ production were measured in Ag+rIL-18 cultures from infected pigs compared to the background response in PBS+rIL-18 control samples (p<0.01) or to Ag+rIL-18 cultures from non-inoculated control pigs (p<0.05). Flow cytometry identified two lymphocyte subsets as the Ag-specific IFN-γ producers. The highest IFN-γ production was by CD4(+)CD8(+) cells while a more numerous population of CD4(-)CD8(+) cells produced lower amounts of IFN-γ in response to rIL-18 and L. intracellularis Ag.     

Primary infection protects pigs against re-infection with Lawsonia intracellularis in experimental challenge studies

In two separate trials pigs were experimentally infected with Lawsonia intracellularis at 5-6 weeks of age followed by antibiotic treatment and resolution of the primary infection and then re-inoculated at 12-13 weeks of age. A treatment-control group of pigs received the primary infection and antibiotic treatment only, and served as control for the antibiotic treatment of the primary infection. A challenge-control group of pigs received the second inoculation dose only at 12-13 weeks of age to control infectivity of the challenge-dose and susceptibility of pigs to L. intracellularis at this age. Pigs were monitored for shedding of L. intracellularis in faeces by PCR, and for the development of antibodies and responses of acute phase proteins in serum. The presence of L. intracellularis antigen in the intestinal mucosa was examined in post mortem samples by immunohistochemistry. In both trials primary infected pigs were protected from infection after challenge inoculation as evidenced by absence of faecal shedding of L. intracellularis, lack of changes in acute phase protein concentrations after challenge and with low levels of bacterial antigen in the intestinal mucosa of re-inoculated pigs comparable to that of the treatment-control pigs. In contrast, challenge-control pigs shed L. intracellularis in faeces, had L. intracellularis antigen extensively present within all layers of the intestinal mucosa and developed a significant acute phase protein response in serum after the experimental infection. The acute phase protein response to L. intracellularis infection was detected as an increased rise in the serum concentrations of C-reactive protein and haptoglobin from day-6 post infection, and increased serum concentrations of haptoglobin were generally seen 2-3 weeks after inoculation both at 5-6 and 12-13 weeks of age. In conclusion substantial protection against L. intracellularis infection was found in the re-inoculated pigs in contrast to the development of infection in age-matched control pigs. The acute phase protein responses reflected both the observed protection against L. intracellularis infection upon secondary challenge and that increased resistance to the infection develops with age.     

A common variant in the FTO gene is associated with body mass index and predisposes to childhood and adult obesity

Obesity is a serious international health problem that increases the risk of several common diseases. The genetic factors predisposing to obesity are poorly understood. A genome-wide search for type 2 diabetes-susceptibility genes identified a common variant in the FTO (fat mass and obesity associated) gene that predisposes to diabetes through an effect on body mass index (BMI). An additive association of the variant with BMI was replicated in 13 cohorts with 38,759 participants. The 16% of adults who are homozygous for the risk allele weighed about 3 kilograms more and had 1.67-fold increased odds of obesity when compared with those not inheriting a risk allele. This association was observed from age 7 years upward and reflects a specific increase in fat mass.     

Potential of dry fractionation of wheat bran for the development of food ingredients,part I: Influence of ultra-fine grinding

Wheat bran is an undervalued by-product of white flour and has great nutritional potential due to its high content in fibres and bioactive compounds. Micronized bran could be used as a food ingredient to improve the nutritional potential of cereal products, or be used as a starting material for other processes (bioactive compound extraction or bran fractionation). The aim of this work was to find a way to efficiently decrease the particle size of bran. The influence of the grinding temperature (ambient or cryogenic grinding) on the granulometric distribution of particles, their composition, and their microstructure was studied, at lab-scale and pilot-scale. It showed that the intrinsic characteristics of bran (glass transition within intermediate layers at −46 °C) had more influence on its grinding behaviour than the type of grinding device used: the particles size distributions obtained after grinding at lab-scale and pilot-scale were very similar. At both scales, the granulometric curves were narrow for cryogenic grinding, while for ambient grinding they were spread over the whole particle size range. Ultrafine particles were obtained in both ambient and cryogenic conditions. Negative temperatures, by increasing the material’s brittleness, favoured a fast fragmentation of bran: one step of cryogenic grinding allowed a median particle size of nearly 50 μm to be reached, whereas three successive steps of ambient grinding were needed for the same result. On the other hand, ambient temperature favoured the dissociation of the different constituent layers of wheat bran, and produced less composite particles than cryogenic grinding.     

BMAA Inhibits Nitrogen Fixation in the Cyanobacterium Nostoc sp. PCC 7120

Cyanobacteria produce a range of secondary metabolites, one being the neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA), proposed to be a causative agent of human neurodegeneration. As for most cyanotoxins, the function of BMAA in cyanobacteria is unknown. Here, we examined the effects of BMAA on the physiology of the filamentous nitrogen-fixing cyanobacterium Nostoc sp. PCC 7120. Our data show that exogenously applied BMAA rapidly inhibits nitrogenase activity (acetylene reduction assay), even at micromolar concentrations, and that the inhibition was considerably more severe than that induced by combined nitrogen sources and most other amino acids. BMAA also caused growth arrest and massive cellular glycogen accumulation, as observed by electron microscopy. With nitrogen fixation being a process highly sensitive to oxygen species we propose that the BMAA effects found here may be related to the production of reactive oxygen species, as reported for other organisms.     

Influence of macronutrient composition,TIBA and dark treatment on shoot formation and nitrogen content in petiole explants of Senecio × hybridus

Summary

The content of ammonium, nitrate and potassium was varied in the macronutrient solutions intended for formation of adventitious shoots from petiole expiants of Senecio × hybridus. The other components of the macronutrients were according to Murashige and Skoog (1962). The largest number of expiants which formed shoots was obtained when the nitrogen concentration in the Murashige-Skoog solution was lowered from 60 to 30 mM and the potassium concentration from 20 to 15 mM. Addition of 1.0 μM TIBA to the medium as well as the standard addition of 4.44 μM BAP and 28.5 μM IAA favoured shoot formation. Even growth in darkness for two weeks immediately after expiant excision increased shoot number. The nitrogen content in the tissue decreased as the nitrogen concentration in the medium decreased, although an increased concentration in the medium from 60 to 75 mM did not increase the nitrogen content in the tissue. When the potassium concentration was changed from 20 to 15 mM, in a medium with 30 mM nitrogen, the nitrogen concentration in the tissue increased. On the other hand, when using a medium with 60 mM nitrogen, the potassium concentration (30 and 20 mM) did not affect the nitrogen content of the tissue.     

Bromination of different morphological parts of spruce wood (Picea abies)

Summary It has been found that the lignin in a middle lamella fraction isolated by a sieving technique has the same reactivity to bromine as the whole wood lignin. A plataue value of 1 mole bromine/mole C₉-unit was obtained for both samples. The lignin in compression wood had a considerably lower reactivity to bromine than the normal wood lignin. A plataue value of 0.68 mole bromine/mol C₉-unit was obtained with such lignin. The discrepancy between this and earlier results from the bromination of middle lamella and the implication of the results for lignin determination by the SEM- and TEM-EDXA technique are discussed.     

Growth and nutrient uptake of ectomycorrhizal Pinus sylvestris seedlings in a natural substrate treated with elevated Al concentrations

Models of the effects of elevated concentrations of aluminum (Al) on growth and nutrient uptake of forest trees frequently ignore the effects of mycorrhizal fungi. In this study, we present novel data indicating that ectomycorrhizal mycelia may prevent leaching of base cations and Al. Mycorrhizal and non-mycorrhizal Pinus sylvestris L. seedlings were grown in sand obtained from the B-horizon of a local forest. In Experiment 1, non-mycorrhizal seedlings and seedlings inoculated with Hebeloma cf. longicaudum (Pers.: Fr.) Kumm. ss. Lange or Laccaria bicolor (Maire) Orton were provided with nutrient solution containing 2.5 mM Al. Aluminum did not affect growth of non-mycorrhizal seedlings or seedlings inoculated with L. bicolor. Seedlings colonized by H. cf. longicaudum had the highest biomass production of all seedlings grown without added Al, but the fungus did not tolerate Al. Shoots of seedlings colonized by L. bicolor had the lowest nitrogen (N) concentrations but the highest phosphorus (P) concentrations of all seedlings. The treatments had small but significant effects on shoot and root Al concentrations. In Experiment 2, inoculation with L. bicolor was factorially combined with the addition of a complete nutrient solution, or a solution lacking the base cations K, Ca and Mg, and solutions containing 0 or 0.74 mM Al. Seedling growth decreased in response to 0.74 mM Al, but the effect was significant only for non-mycorrhizal seedlings. Mycorrhizal seedlings generally had higher P concentrations than non-mycorrhizal seedlings. Aluminum reduced P uptake in non-mycorrhizal plants but had no effect on P uptake in mycorrhizal plants. Mycorrhizal colonization increased the pH of the soil solution by about 0.5 units and addition of Al decreased the pH by the same amount. We conclude that the presence of ectomycorrhizal mycelia decreased leaching of base cations and Al from the soil.     

Effect of different water supply on plant growth and fruit quality of Lycopersicon esculentum

It is well known from earlier work that water stress and salinity results in depressed plant growth and high fruit quality of tomato (e.g. increased sugar and acid levels), but generally is associated with a low marketable fruit yield. In the present work we investigated whether even a small reduction in water supply (without visible symptoms of water stress) also results in a high fruit quality together with high marketable fruit proportions. To characterize fruit quality sugars (glucose and fructose), titratable acids, odour-active aroma volatiles and vitamin C were investigated. Tomato plants (Lycopersicon esculentum Mill. cv Vanessa) were grown in soil and with the onset of fruit development water supply was varied (70% and 50% water capacity). In the treatment with lower water supply plant growth, and in particular the number of fruit settings were depressed and the sugar and vitamin C concentrations in the fruits were significantly increased, especially during fruit ripening. Furthermore, with lower water supply the concentrations of titratable acids and of C₆ aldehydes (hexanal, (Z)-3-hexenal and (E)-2-hexenal) were significantly increased in the red fruits. Fruit growth was identical in both treatments. The higher levels of sugars, titratable acids, aroma volatiles and vitamin C are responsible for the higher fruit quality under conditions of lower water supply. Since not all fruits of the well watered plants became mature, the marketable yield in both treatments was rather similar and hence, together with a higher fruit quality in the treatment with lower water supply, high proportions of marketable fruits can be harvested.