Nitro-oleic acid regulates T cell activation through post-translational modification of calcineurin.

Nitro-fatty acids (NO2-FAs) are unsaturated fatty acid nitration products that exhibit anti-inflammatory actions in experimental mouse models of autoimmune and allergic diseases. These electrophilic molecules interfere with intracellular signaling pathways by reversible post-translational modification of nucleophilic amino-acid residues. Several regulatory proteins have been identified as targets of NO2-FAs, modifying their activity and promoting gene expression changes that result in anti-inflammatory effects. Herein, we report the effects of nitro-oleic acid (NO2-OA) on pro-inflammatory T cell functions, showing that 9- and 10-NOA, but not their oleic acid precursor, decrease T cell proliferation, expression of activation markers CD25 and CD71 on the plasma membrane, and IL-2, IL-4, and IFN-γ cytokine gene expressions. Moreover, we have found that NO2-OA inhibits the transcriptional activity of nuclear factor of activated T cells (NFAT) and that this inhibition takes place through the regulation of the phosphatase activity of calcineurin (CaN), hindering NFAT dephosphorylation, and nuclear translocation in activated T cells. Finally, using mass spectrometry-based approaches, we have found that NO2-OA nitroalkylates CaNA on four Cys (Cys129, 228, 266, and 372), of which only nitroalkylation on Cys372 was of importance for the regulation of CaN phosphatase activity in cells, disturbing functional CaNA/CaNB heterodimer formation. These results provide evidence for an additional mechanism by which NO2-FAs exert their anti-inflammatory actions, pointing to their potential as therapeutic bioactive lipids for the modulation of harmful T cell-mediated immune responses.

Biochar reduces volatile organic compounds generated during chicken manure composting.

The efficiency of biochar for reducing the levels of volatile organic compounds (VOC) was investigated in a composting mixture containing 90% poultry manure and 10% straw (with and without 3% biochar addition) at three different stages of the process. The use of a low application rate of biochar reduced the concentration of VOC during the thermophilic phase. Biochar significantly reduced the levels of nitrogen volatile compounds, which are the most abundant VOC family, originated from microbial transformation of the N-compounds originally present in manure. The most efficient VOC reduction was observed in oxygenated volatile compounds (ketones, phenols and organic acids), which are intermediates of organic matter degradation, whereas there was no effect on other VOC families (aliphatic, aromatic and terpenes). These results suggest the importance of not only the sorption capacity of biochar but also its impact in the composting progress as main drivers for VOC reduction.

Biochar reduces the efficiency of nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) mitigating N2O emissions.

Among strategies suggested to decrease agricultural soil N2O losses, the use of nitrification inhibitors such as DMPP (3,4-dimethylpyrazole phosphate) has been proposed. However, the efficiency of DMPP might be affected by soil amendments, such as biochar, which has been shown to reduce N2O emissions. This study evaluated the synergic effect of a woody biochar applied with DMPP on soil N2O emissions. A incubation study was conducted with a silt loam soil and a biochar obtained from Pinus taeda at 500 °C. Two biochar rates (0 and 2% (w/w)) and three different nitrogen treatments (unfertilized, fertilized and fertilized + DMPP) were assayed under two contrasting soil water content levels (40% and 80% of water filled pore space (WFPS)) over a 163 day incubation period. Results showed that DMPP reduced N2O emissions by reducing ammonia-oxidizing bacteria (AOB) populations and promoting the last step of denitrification (measured by the ratio nosZI + nosZII/nirS + nirK genes). Biochar mitigated N2O emissions only at 40% WFPS due to a reduction in AOB population. However, when DMPP was applied to the biochar amended soil, a counteracting effect was observed, since the N2O mitigation induced by DMPP was lower than in control soil, demonstrating that this biochar diminishes the efficiency of the DMPP both at low and high soil water contents.

Relationships between emitted volatile organic compounds and their concentration in the pile during municipal solid waste composting.

Composting operations taking place at municipal solid waste (MSW) treatment plants represent a source of volatile organic compounds (VOC) to the atmosphere. Understanding the variables governing the release of VOC at these facilities is crucial to assess potential health risks for site workers and local residents. In this work the changes in the VOC composition of a composting pile were monitored and compared to the VOC emmited from the same pile in order to understand the impact of composting operations on the release of VOC. More than one hundred VOC were indentified in the solid phase of the composting piles, which were dominated by terpenes (about 50% of the total amount of VOC) and in a lower quantity alcohols, volatile fatty acids and aromatic compounds. There was a reduction in the total concentration of VOC in the pile during composting, from 45 to 35 mg/kg, but the compostion and distribution of VOC families remained stable in the pile even in the mature compost. However, there was no correlation between the emitted VOC and their concentration in the composting pile. The VOC emission pattern was affected by the biological activity in the pile (measured by temperature, CO2 evolution and the presence of CH4 emissions). The highest VOC emissions were detected at early stages of the process, alongside with the generation of CH4 in the pile, and then decreased sharply in the mature compost as a consequence of biodegradation and volatilisation. These results pointed to the importance of composting operation rather than the composition of the raw materials on the release of VOC in composting plants.

Role of biochar as an additive in organic waste composting.

The use of biochar in organic waste composting has attracted interest in the last decade due to the environmental and agronomical benefits obtained during the process. Biochar presents favourable physicochemical properties, such as large porosity, surface area and high cation exchange capacity, enabling interaction with major nutrient cycles and favouring microbial growth in the composting pile. The enhanced environmental conditions can promote a change in the microbial communities that can affect important microbially mediated biogeochemical cycles: organic matter degradation and humification, nitrification, denitrification and methanogenesis. The main benefits of the use of biochar in composting are reviewed in this article, with special attention to those related to the process performance, compost microbiology, organic matter degradation and humification, reduction of N losses and greenhouse gas emissions and fate of heavy metals.

Biochar accelerates organic matter degradation and enhances N mineralisation during composting of poultry manure without a relevant impact on gas emissions.

A composting study was performed to assess the impact of biochar addition to a mixture of poultry manure and barley straw. Two treatments: control (78% poultry manure + 22% barley straw, dry weight) and the same mixture amended with biochar (3% dry weight), were composted in duplicated windrows during 19 weeks. Typical monitoring parameters and gaseous emissions (CO2, CO, CH4, N2O and H2S) were evaluated during the process as well as the agronomical quality of the end-products. Biochar accelerated organic matter degradation and ammonium formation during the thermophilic phase and enhanced nitrification during the maturation phase. Our results suggest that biochar, as composting additive, improved the physical properties of the mixture by preventing the formation of clumps larger than 70 mm. It favoured microbiological activity without a relevant impact on N losses and gaseous emissions. It was estimated that biochar addition at 3% could reduce the composting time by 20%.

High concentrations of polycyclic aromatic hydrocarbons (naphthalene, phenanthrene and pyrene) failed to explain biochar's capacity to reduce soil nitrous oxide emissions.

The presence of polycyclic aromatic hydrocarbons (PAHs) has been postulated as a mechanism by which biochar might mitigate N(2)O emissions. We studied whether and to what extent N(2)O emissions were influenced by the three most abundant PAHs in biochar: naphthalene, phenanthrene and pyrene. We hypothesised that biochars contaminated with PAHs would show a larger N(2)O mitigation capacity and that increasing PAH concentrations in biochar would lead to higher mitigation potentials. Our results demonstrate that the high-temperature biochar (550 °C) had a higher capacity to mitigate soil N(2)O emissions than the low-temperature biochar (350 °C). At low PAH concentrations, PAHs do not significantly contribute to the reductions in soil N(2)O emissions; while biochar stimulated soil N(2)O emissions when it was spiked with high concentrations of PAHs. This study suggests that the impact of biochar on soil N(2)O emissions is due to other compositional and/or structural properties of biochar rather than to PAH concentration.

P2X(7) receptor activation enhances SK3 channels- and cystein cathepsin-dependent cancer cells invasiveness.

ATP-gated P2X(7) receptors (P2X(7)R) are unusual plasma membrane ion channels that have been extensively studied in immune cells. More recently, P2X(7)R have been described as potential cancer cell biomarkers. However, mechanistic links between P2X(7)R and cancer cell processes are unknown. Here, we show, in the highly aggressive human breast cancer cell line MDA-MB-435s, that P2X(7) receptor is highly expressed and fully functional. Its activation is responsible for the extension of neurite-like cellular prolongations, of the increase in cell migration by 35% and in cell invasion through extracellular matrix by 150%. The change in cancer cell morphology and the increased migration appeared to be due to the activation of Ca(2+)-activated SK3 potassium channels. The enhanced invasion through the extracellular matrix was related to the increase of mature forms of cysteine cathepsins in the extracellular medium, which was independent of SK3 channel activity and not associated with cell death. Pharmacological targeting of P2X(7)R in vivo was crucial for cell invasiveness in a zebrafish model of metastases. Our results demonstrate a novel mechanistic link between P2X(7)R functionality in cancer cells and invasiveness, a key parameter in tumour growth and in the development of metastases. They also suggest a potential therapeutic role for the newly developed P2X(7)R antagonists.

Potential of olive mill waste and compost as biobased pesticides against weeds, fungi, and nematodes.

The phytotoxic and antimicrobial properties of olive mill wastes have been widely investigated and demonstrated over the past decade. However, their potential utilization as biodegradable pesticides against plant pathogens is still poorly understood. In this study, a series of laboratory bioassays was designed to test the inhibitory effects of sterile water extracts of two-phase olive mill waste (TPOMW) and TPOMW composts with different degrees of stabilization on several different plant pathogens. Fungicidal properties of TPOMW extracts, assayed in a microwell assay format, showed that the growth of Phytophthora capsici was consistently and strongly inhibited by all TPOMW extracts diluted 1:10 (w:v). In contrast, suppression of Pythium ultimum and Botrytis cinerea by the extracts was not as strong and depended on the specific TPOMW sample. Mature compost inhibited P. capsici and B. cinerea at dilutions as great as 1:50, w:v. Neither TPOMW nor TPOMW compost extracts were able to inhibit the growth of the basidiomycete root rot agent Rhizoctonia solani. In addition, studies were conducted on the allelopathic effects of TPOMW extracts on seed germination of four highly invasive and globally distributed weeds (Amaranthus retroflexus, Solanum nigrum, Chenopodium album and Sorghum halepense). Both the TPOMW and immature TPOMW compost extracts substantially inhibited germination of A. retroflexus and S. nigrum, whereas mature composts extracts only partially reduced the germination of S. nigrum. Finally, TPOMW extracts strongly inhibited egg hatch and second-stage juvenile (J2) motility of the root-knot nematode Meloidogyne incognita. However, only higher concentrations of stage-one and stage-two TPOMW compost extracts exerted a suppressive effect on both J2 motility and on egg hatch. The study shows the high potential of naturally occurring chemicals present in TPOMW and TPOMW composts that should be further investigated as bio-pesticides for their use in sustainable agricultural systems.

The mineralisation of fresh and humified soil organic matter by the soil microbial biomass.

Soil organic matter comprises all dead plant and animal residues, from the most recent inputs to the most intensively humified. We have found that traces of fresh substrates at microg g(-1) soil concentrations (termed 'trigger molecules') activate the biomass to expend more energy than is contained in the original 'trigger molecules'. In contrast, we suggest that the rate limiting step in soil organic matter mineralisation is independent of microbial activity, but is governed by abiological processes (which we term the Regulatory Gate theory). These two findings have important implications for our understanding of carbon mineralisation in soil, a fundamental process in the sequestration of soil organic matter.

Carbon mineralization dynamics in soils amended with meat meals under laboratory conditions.

Meat and bone meal (MBM) is obtained from the wastes produced during slaughtering operations. Its high concentration of N and P makes it interesting as an organic fertiliser but its use in soil has been barely studied previously. In this work four laboratory experiments were performed to study the influence of different variables (MBM composition, rate of application, temperature of incubation and the type of soil) on C mineralization dynamics of MBM in agricultural soils. The total CO2-C evolved (as % of added C) after 2 weeks ranged between 10% and 20%. The kinetics of mineralization were rapid, with C evolved as CO2 within the first 4 days representing more than 50% of total C mineralized. A linear correlation was found between the rate of application (added-C) and CO2-C evolved (r2: 0.997; P<0.001). A temperature coefficient (Q10) was used to assess the difference in biological activity at 5 degrees C intervals. Q10, which ranged from 1.0 to 2.7 (250h), was higher for the lower temperature range (Q10 (15-20 degrees C)>Q10 (20-25 degrees C)) and it was found to be related to the soil properties. Finally, the mineralization process was found to be highly dependent upon the different soil factors, although no simple linear correlation was found between mineralization and soil properties.

Potential of olive mill wastes for soil C sequestration.

The present work deals with the potential of olive mill wastes as a C source for soil C sequestration strategy, which is based on the high lignocellulosic content that makes these wastes to degrade slowly during composting and after land application. A C balance was performed during the whole life cycle of two different two-phase olive mill wastes (TPOMW): C losses were calculated during the composting process and after soil application of the composting mixtures under laboratory conditions. The effect of the degree of stabilization of TPOMW on the overall C waste conservation efficiency was also evaluated. C losses after 34 weeks of TPOMW composting ranged from 40.58% to 45.19% of the initial C, whereas the amount of C evolved as CO2 after 8 months of incubation of soil amended with mature composts only represented between 20.6% and 21.9% of the added C. The total C losses considering the whole life cycle of the TPOMW showed lower losses compared to composts prepared with organic residues of different origin. Conversely to the typical behaviour of other organic wastes, the stabilisation degree of the TPOMW composting mixtures did not show any significant effect on the total C losses measured during composting and later land application. The low rate of degradation of TPOMW both during composting and after soil application makes the use of TPOMW as a C source an attractive strategy for soil C sequestration.

Chemical properties and hydrolytic enzyme activities for the characterisation of two-phase olive mill wastes composting.

Two-phase olive mill waste (TPOMW) is a semisolid sludge generated during the extraction of olive oil by the two-phase centrifugation system. Among all the available disposal options, composting is gaining interest as a sustainable strategy to recycle TPOMW for agricultural purposes. The quality of compost for agronomical use depends on the degree of organic matter stabilization, but despite several studies on the topic, there is not a single method available which alone can give a certain indication of compost stability. In addition, information on the biological and biochemical properties, including the enzymatic activity (EA) of compost, is rare. The aim of this work was to investigate the suitability of some enzymatic activities (beta-glucosidase, arylsulphatase, acid-phosphatase, alkaline-phosphatase, urease and fluorescein diacetate hydrolysis (FDA)) as parameters to evaluate organic matter stability during the composting of TPOMW. These enzymatic indices were also compared to conventional stability indices. For this purpose two composting piles were prepared by mixing TPOMW with sheep manure and grape stalks in different proportions, with forced aeration and occasional turnings. The composting of TPOMW followed the common pattern reported previously for this kind of material with a reduction of 40-50% of organic matter, a gradual increase in pH, disappearance of phytotoxicity and formation of humic-like C. All EA increased during composting except acid-phosphatase. Significant correlations were found between EA and some important conventional stability indices indicating that EA can be a simple and reliable tool to determine the degree of stability of TPOMW composts.

Duckweed (Lemna gibba) growth inhibition bioassay for evaluating the toxicity of olive mill wastes before and during composting.

Two-phase olive mill waste (TPOMW) is considered the main problem confronting the modern oil extraction and processing industry. Composting has been recently proposed as a suitable method to treat TPOMW so that it is suitable for use in agriculture. In the work reported here, the Lemna gibba bioassay was tested to assess the toxicity of TPOMW before and during the composting process. The method was compared with the Lepidium sativum bioassay and with other chemical maturity indices traditionally reported in the literature. The L. gibba test proved to be a simple, sensitive, and accurate method to evaluate toxicity before and during the composting of TPOMW. Plant growth response was measured by two methods: counting the number of fronds (leaves) and measuring total frond area (TFA) with image analysis software. Compared to the counting of fronds (L. gibba) or seeds (L. sativum), the use of area-measuring software permitted a very rapid, unbiased and easy way of analysing the toxicity of TPOMW before and during composting. Although the accuracy of the frond count method was similar to the traditional cress seed test, data analysis showed that the TFA measurement method was statistically more accurate (significantly lower variance) than the frond count approach. Highly significant correlations were found between TFA and some important maturation indices commonly reported in literature indicating that the L. gibba bioassay can be a useful tool to determine the degree of maturity of TPOMW composts.

Influence of fungicides on grape yeast content and its evolution in the fermentation.

The influence of six fungicides (famoxadone, fenhexamid, fluquinconazole, kresoxim-methyl, quinoxyfen, trifloxystrobin) on the yeast content in harvested grapes has been studied along with their effect on it during the wine-making process. Two treatments were carried out with authorized formulates at the manufacturer doses. The first was carried out under good agricultural practices, obeying the security times, and the second one under critical conditions, applied on the day of harvesting. The grapes were harvested two hours after the application. During the wine-making process, samples were taken at 1, 5, 12 and 20 days after the start. The levels observed in the control sample (1.9 log CFU/cm2) were similar to previous studies. The counts in all treated samples were higher than the control sample, without adverse effect of the pesticides on the yeast count even in the grapes treated on the day of harvest. As with the observations on grapes, no effect of the pesticides on the fermentation count was observed. All counts on the treated samples were higher than the control sample. The treatment on the day of harvest did not have any effect on the fermentation count, with superior results being obtained for all the pesticides.

An overview on olive mill wastes and their valorisation methods.

Olive mill wastes represent an important environmental problem in Mediterranean areas where they are generated in huge quantities in short periods of time. Their high phenol, lipid and organic acid concentrations turn them into phytotoxic materials, but these wastes also contain valuable resources such as a large proportion of organic matter and a wide range of nutrients that could be recycled. In this article, recent research studies for the valorisation of olive mill wastes performed by several authors were reviewed: second oil extraction, combustion, gasification, anaerobic digestion, composting and solid fermentation are some of the methods proposed. Special attention was paid to the new solid waste generated during the extraction of olive oil by the two-phase system. The peculiar physicochemical properties of the new solid waste, called two-phase olive mill waste, caused specific management problems in the olive mills that have led to the adaptation and transformation of the traditional valorisation strategies. The selection of the most suitable or appropriate valorisation strategy will depend on the social, agricultural or industrial environment of the olive mill. Although some methods are strongly consolidated in this sector, other options, more respectful with the environment, should also be considered.

The use of elemental sulphur as organic alternative to control pH during composting of olive mill wastes.

High values of pH may represent a limitation for the agricultural use of the composts, not only when used as soil-less substrate but also as soil amendment in high pH soils. The addition of elemental S during the maturation phase of the composting process was evaluated as suitable method to reduce pH of the composts under the organic agriculture regulations. A compost prepared with two phase olive mill waste (OMW) and sheep litter (SL) was used to study the effect of elemental sulphur addition on the pH of the composting mixture. Initially, different bench scale experiments were designed in order to study the influence of moisture, sulphur concentration, and incubation temperature on the sulphur oxidation rate and thus on the pH of the compost. A concentration of 0.5% in sulphur (dry weight basis) and moisture of 40% were proposed as the optimum conditions to decrease the compost pH by 1.1 units without increasing in EC to levels that may suppose a limitation for its agricultural use. Finally, these optimum experimental conditions found at bench scale were tested at full scale in a commercial composting plant treating the same organic materials by windrowing. The pH values of the composting mixture were reduced by one unit after 2 weeks following the addition of elemental S causing no negative effects on the final compost quality.

Germline mutational dynamics in myotonic dystrophy type 1 males: allele length and age effects.

BACKGROUND: The CTG repeat expansion causing myotonic dystrophy type 1 is unstable in the germline, and frequent intergenerational length changes are observed, giving rise to the unusual genetics of the disorder. The repeat is also somatically unstable, and expanded alleles accumulate throughout life, thus compromising simple measures of intergenerational stability. OBJECTIVE: To gain a better understanding of the intergenerational dynamics of the DM1 repeat in the male germline. METHODS: We used sensitive small pool PCR procedures to analyze sperm and somatic DNA from 22 DM1 men of different ages, CTG repeat length, and clinical form. RESULTS: High levels of repeat length variation heavily biased toward further expansions were observed in the sperm of all DM1 men. Progenitor allele length was revealed as a major modifier of interindividual variation, with the largest length changes observed for premutation and protomutation alleles and the highest frequency of contractions in full mutation alleles. However, despite clear increases in the degree of somatic mosaicism, no differences were observed in replicate sperm samples obtained from two men during a 4-year period. CONCLUSIONS: Progenitor allele length is a major modifier of the mutational dynamics of the DM1 repeat in the male germline, but surprisingly age is not. Therefore, other as yet unidentified modifiers must be responsible for the considerable residual interindividual variation that cannot be accounted for by these factors.