Human Neutrophils Couple Nitric Oxide Production and Extracellular Traps Formation in Allergic Asthma.

RATIONALE: Nitric oxide (NO) is elevated in the airways and serum of allergic asthmatic patients, suggesting an important role in asthma. NO production has been widely attributed to the canonical inducible nitric oxide synthase (iNOS). Much effort has been made to inhibit this enzyme with two outcomes: no asthma improvement; and partial NO reduction, suggesting the involvement of an iNOS-independent source. OBJECTIVES: Neutrophils produce NO under inflammatory conditions and their role in asthma has been overlooked. The present study analyzes their possible role as source of NO. METHODS: Our hypothesis was tested in 99 allergic patients with intermittent bronchial asthma and 26 healthy donors. NO production by blood and sputum neutrophils in response to allergens, anti-IgE, and anti-IgE receptors Abs was assessed by Griess, flow cytometry and confocal microscopy. Extracellular traps (ETs) formation, as a possible consequence of NO production, was quantified by western blot and confocal microscopy, and reactive oxygen species by luminol-enhanced chemiluminescence. RESULTS: Among blood and sputum granulocytes from allergic asthmatic patients, only neutrophils, produce NO by an IgE-dependent mechanism. This production is independent of NOS, but dependent on a reaction between L-arginine and reactive oxygen species from NOX2. NO and ETosis are induced in parallel, and NO amplifies ETs formation, which is a key mediator in asthma. CONCLUSIONS: Our findings reveal a novel role of neutrophils as the unique allergen/IgE-dependent NO source in allergic asthma enhancing ETs formation. These results suggest that NO produced by neutrophils needs further consideration in the treatment of allergic asthma.

Grazing livestock move by Lévy walks: Implications for soil health and environment.

Grazing livestock plays an important role in the context of food security, agricultural sustainability and climate change. Understanding how livestock move and interact with their environment may offer new insights on how grazing practices impact soil and ecosystem functions at spatial and temporal scales where knowledge is currently limited. We characterized daily and seasonal grazing patterns using Global Positioning System (GPS) data from two grazing strategies: conventionally- and rotationally-grazed pastures. Livestock movement was consistent with the so-called Lévy walks, and could thus be simulated with Lévy-walk based probability density functions. Our newly introduced "Moovement model" links grazing patterns with soil structure and related functions by coupling animal movement and soil structure dynamics models, allowing to predict spatially-explicit changes in key soil properties. Predicted post-grazing management-specific bulk densities were consistent with field measurements and confirmed that rotational grazing produced similar disturbance as conventional grazing despite hosting higher stock densities. Harnessing information on livestock movement and its impacts in soil structure within a modelling framework can help testing and optimizing grazing strategies for ameliorating their impact on soil health and environment.

Response of soil health indicators to dung, urine and mineral fertilizer application in temperate pastures.

Healthy soils are key to sustainability and food security. In temperate grasslands, not many studies have focused on soil health comparisons between contrasting pasture systems under different management strategies and treatment applications (e.g. manures and inorganic fertilisers). The aim of this study was to assess the responses of soil health indicators to dung, urine and inorganic N fertiliser in three temperate swards: permanent pasture not ploughed for at least 20 years (PP), high sugar ryegrass with white clover targeted at 30% coverage reseeded in 2013 (WC), and high sugar ryegrass reseeded in 2014 (HG). This study was conducted on the North Wyke Farm Platform (UK) from April 2017 to October 2017. Soil health indicators including soil organic carbon (SOC, measured by loss of ignition and elemental analyser), dissolved organic carbon (DOC), total nitrogen (TN), C:N ratio, soil C and N bulk isotopes, pH, bulk density (BD), aggregate stability, ergosterol concentration (as a proxy for fungi biomass), and earthworms (abundance, mass and density) were measured and analysed before and after application of dung and N fertilizer, urine and N fertiliser, and only N fertiliser. The highest SOC, TN, DOC, ergosterol concentration and earthworms as well as the lowest BD were found in PP, likely due to the lack of ploughing. Differences among treatments were observed due to the application of dung, resulting in an improvement in chemical indicators of soil health after 50 days of its application. Ergosterol concentration was significantly higher before treatment applications than at the end of the experiment. No changes were detected in BD and aggregate stability after treatment applications. We conclude that not enough time had passed for the soil to recover after the ploughing and reseeding of the permanent pasture, independently of the sward composition (HG or WC). Our results highlight the strong influence of the soil management legacy in temperate pasture and the positive effects of dung application on soil health over the short term. In addition, we point out the relevance of using standardised methods to report soil health indicators and some methodological limitations.

GeoHealth: Geographic Information System for Health Management and Clinical, Epidemiological and Translational Research.

This project aims to develop a system for clinical, epidemiological and translational research capable of associating contextual variables and geospatial data with clinical patient information. The GeoHealth system will include a section to perform exploratory analysis that will help identify risk factors to optimize clinical decision making.

Empiric antibiotics for community-acquired pneumonia in adult patients: a systematic review and a network meta-analysis.

OBJECTIVE: The main aim of this network meta-analysis is to identify the empiric antibiotic (Em-ATB) with the highest probability of being the best (HPBB) in terms of (1) cure rate and (2) mortality rate in hospitalised patients with community acquired pneumonia (CAP) . METHOD: Inclusion criteria: (1) adult patients (>16 years old) diagnosed with CAP that required hospitalisation; (2) randomised to at least two different Em-ATBs, (3) that report cure rate and (4) are written in English or Spanish. EXCLUSION CRITERIA: (1) ambiguous antibiotics protocol and (2) published exclusively in abstract or letter format. DATA SOURCES: Medline, Embase, Cochrane and citation reviews from 1 January 2000 to 31 December 2018. Risk of bias: Cochrane's tool. Quality of the systematic review (SR): A MeaSurement Tool to Assess systematic Reviews-2. Certainity of the evidence: Grading of Recommendations Assessment, Development and Evaluation. STATISTICAL ANALYSES: frequentist method performed with the 'netmeta' library, R package. RESULTS: 27 randomised controlled trials (RCTs) from the initial 41 307 screened citations were included. Regarding the risk of bias, more than one quarter of the studies presented low risk and no study presented high risk in all domains. The SR quality is moderate. For cure, two networks were constructed. Thus, two Em-ATBs have the HPBB: cetaroline 600 mg (two times a day) and piperacillin 2000 mg (two times a day). For mortality, three networks were constructed. Thus, three Em-ATBs have the HPBB: ceftriaxone 2000 mg (once a day) plus levofloxacin 500 (two times a day), ertapenem 1000 mg (two times a day) and amikacin 250 mg (two times a day) plus clarithromycin 500 mg (two times a day). The certainity of evidence for each results is moderate. CONCLUSION: For cure rate, ceftaroline and piperaciline are the options with the HPBB. However, for mortality rate, the options are ceftriaxone plus levofloxacin, ertapenem and amikacin plus clarithromycin. It seems necessary to conduct an RCT that compares treatments with the HPBB for each event (cure or mortality) (CRD42017060692).

Implications of afforestation vs. secondary succession for soil properties under a semiarid climate.

Afforestation or secondary succession after cropland abandonment are different strategies to restore soil ecosystem services such as nutrient cycling, soil conservation, and carbon sequestration. However, the studies on the effects on soil-property dynamics after land-use changes are limited in semiarid regions. In this context, an experimental area with a semiarid climate allowed the assessment and comparison of physicochemical soil properties (soil organic C [SOC], soil total N [TN], available P [AP], available K [AK], cation-exchange capacity [CEC], bulk density [BD], pH, available water-holding capacity [AWHC], and C:N ratio) after Pinus halepensis afforestation and secondary succession following agriculture abandonment in 1994. The impact of 12 soil-preparation treatments for planting on soil properties was also evaluated. For this, soil samples (0-10 cm deep) from the afforestation were taken in 1998, 2002, 2007, 2010, 2013, and 2016, and from abandoned cropland in 2010, 2013, and 2016. In afforestation, soil-preparation treatments did not alter differentially the soil properties after 22 years. Significant differences among years were found in SOC, TN, AP, AK, CEC, pH, and C:N. BD changes were detected neither in afforestation nor in abandoned cropland. After 22 years, only SOC, AK and the C:N ratio proved significantly higher in afforestation than in abandoned cropland. In general, soil properties improvement (i.e. SOC, TN, AP, AK, and CEC) was slow after afforestation and abandoned cropland likely due to the legacy of the previous land use (cereal crops) and the semiarid climate influence.

Distinct triterpene synthases in the laticifers of Euphorbia lathyris.

Euphorbia lathyris was proposed about fifty years ago as a potential agroenergetic crop. The tremendous amounts of triterpenes present in its latex has driven investigations for transforming this particular biological fluid into an industrial hydrocarbon source. The huge accumulation of terpenes in the latex of many plant species represent a challenging question regarding cellular homeostasis. In fact, the enzymes, the mechanisms and the controllers that tune the amount of products accumulated in specialized compartments (to fulfill ecological roles) or deposited at important sites (as essential factors) are not known. Here, we have isolated oxidosqualene cyclases highly expressed in the latex of Euphorbia lathyris. This triterpene biosynthetic machinery is made of distinct paralogous enzymes responsible for the massive accumulation of steroidal and non-steroidal tetracyclic triterpenes. More than eighty years after the isolation of butyrospermol from shea butter (Heilbronn IM, Moffet GL, and Spring FS J. Chem. Soc. 1934, 1583), a butyrospermol synthase is characterized in this work using yeast and in folia heterologous expression assays.

Riesgos psicosociales considerados por jueces de familia en decisiones sobre pérdida de patria potestad: Estudio exploratorio / Psychosocial risks considered by family judges in decisions on loss of parental authority: An exploratory study

La investigación realizada tuvo como objetivo identificar riesgos psicosociales considerados por jueces en decisiones sobre pérdida de la patria potestad. El tipo y diseño de la investigación fue cualitativo. Las fuentes de información eran dos: (1) se preseleccionaron 50 documentos judiciales en las siguientes ciudades de Colombia: Bogotá, Tunja y Duitama y (2) se hizo una entrevista grupal con ocho jueces. Los documentos fueron analizados con un protocolo y para diseñarlo, se determinaron categorías orientadas por los objetivos y sustento teórico. La entrevista consistió en un grupo focal. El audio de la entrevista se transcribió literalmente y se realizó un análisis categorial inductivo. Los hallazgos indican que en la mayor parte de casos, la causal más común corresponde al abandono. Allí se identifica más un interés por viajar con el niño fuera del país sin tener que pedir permiso al progenitor demandado. Igualmente se observa que poco se tiene en cuenta la interdisciplinariedad y el interés superior del menor dentro de las decisiones. En el momento de tomar la decisión los jueces utilizan más la lógica silogística que la realista. Empero, en la discusión generada dentro del contexto de la entrevista grupal, anteponen la experiencia profesional y posiblemente vivencias propias para describir las situaciones evaluadas (lógica realista). De la misma manera, reconocen riesgos psicosociales que no quedan expuestos en el documento judicial. Finalmente, se encuentra la necesidad de evaluar más detalladamente la historia relacional de la pareja parental, ya que suelen existir secuelas psicológicas que afectan las competencias parentales.

Suspension of custody as a right of the child may lead to psychosocial risks considered as potential dangers to the well-being of the child, his family or his community in various dimensions. It is therefore important to consider the possibility of these psychosocial risks when making these judicial decisions about the loss of parental rights. The phenomenon of psychosocial risk has been more studied in working environments and also linked to health problems. This fact assigns a new connotation to this study, since it opens the door to the interdisciplinarity from Psychology and Law, when investigating if the psychosocial risks in the judicial decisions on loss of parental authority are taken into account. The research aims to identify psychosocial risks considered by family judges in decisions about loss of custody. The type and design research is qualitative, since one of its general pretensions is the understanding. The sources of information are two. The first corresponds to 50 pre-selected court documents in the Colombian cities of Bogota, Tunja and Duitama. The other is a group interview with 8 family judges. The documents were analyzed by means of a protocol. For the design and implementation of the protocol, categories oriented by interdisciplinary research team objectives, theoretical support and expectations were made. The interview consisted of a focus group interview with family judges (four men and four women). The audio of the interview was transcribed literally and an inductive categorical analysis was made. Results of the analysis of the interview and documents were contrasted. Judicial documents reviewed show that judges use syllogistic logic when seeking to accommodate situations to the causal. Within the judicial decisions it is observed that the best interest of the minor is not privileged. The analysis of the interview in which the use of experience and intuition is observed following a realistic logic, invites us to reflect on aspects such as: the subjectivity of udges in the evaluation, the pressure they face on the number of lawsuits and the speed of judgment, and the need to revise the causal deprivation of liberty superior to one year. The judges manifest a deficit in the interdisciplinary work and the lack of more training in aspects of mental health that would make possible the use of psychological reports and the greater understanding of causes such as disability. It was possible to identify a pattern that leads to the decision of loss of custody power by abandonment (most common causal): (1) Search for legal advice. (2) To promote the notoriety of the physical and emotional distance between the defendant and the child. (3) Establish the claim. (4) Accumulate evidential evidence on the causal. (5) Obtain witnesses. In the discussion generated in the group interview, judges recognize psychosocial risks that are not exposed in the court document or sentence. Finally there is the need to assess in more detail the relational history of the parental couple, as they tend to be psychological consequences that affect parenting skills. The discussion between the researchers of Family Psychology and those of Family Law appear questions that build an interdisciplinary bridge. One limitation of the study was that it was not possible to have sufficient time and resources to conduct group and individual interviews with a greater number of family judges in other cities of the country and achieve more generalizable results. The research question on whether judges take into account the risks to make the decisions is answered that it attends to the grounds provided in the law taking the decision guided by the syllogistic logic. Causals can be equated with psychosocial risks.

An Oleuropein ß-Glucosidase from Olive Fruit Is Involved in Determining the Phenolic Composition of Virgin Olive Oil.

Phenolic composition of virgin olive oil is determined by the enzymatic and/or chemical reactions that take place during olive fruit processing. Of these enzymes, ß-glucosidase activity plays a relevant role in the transformation of the phenolic glycosides present in the olive fruit, generating different secoiridoid derivatives. The main goal of the present study was to characterize olive fruit ß-glucosidase genes and enzymes responsible for the phenolic composition of virgin olive oil. To achieve that, we have isolated an olive ß-glucosidase gene from cultivar Picual (OepGLU), expressed in Nicotiana benthamiana leaves and purified its corresponding recombinant enzyme. Western blot analysis showed that recombinant OepGLU protein is detected by an antibody raised against the purified native olive mesocarp ß-glucosidase enzyme, and exhibits a deduced molecular mass of 65.0 kDa. The recombinant OepGLU enzyme showed activity on the major olive phenolic glycosides, with the highest levels with respect to oleuropein, followed by ligstroside and demethyloleuropein. In addition, expression analysis showed that olive GLU transcript level in olive fruit is spatially and temporally regulated in a cultivar-dependent manner. Furthermore, temperature, light and water regime regulate olive GLU gene expression in olive fruit mesocarp. All these data are consistent with the involvement of OepGLU enzyme in the formation of the major phenolic compounds present in virgin olive oil.

Exciton Transfer and Emergent Excitonic States in Oppositely-Charged Conjugated Polyelectrolyte Complexes.

Photosynthetic organisms have mastered the use of "soft" macromolecular assemblies for light absorption and concentration of electronic excitation energy. Nature's design centers on an optically inactive protein-based backbone that acts as a host matrix for an array of light-harvesting pigment molecules. The pigments are organized in space such that excited states can migrate between molecules, ultimately delivering the energy to the reaction center. Here we report our investigation of an artificial light-harvesting energy transfer antenna based on complexes of oppositely charged conjugated polyelectrolytes (CPEs). The conjugated backbone and the charged side chains of the CPE lead to an architecture that simultaneously functions as a structural scaffold and an electronic energy "highway". We find that the process of ionic complex formation leads to a remarkable change in the excitonic wavefunction of the energy acceptor, which manifests in a dramatic increase in the fluorescence quantum yield. We argue that the extended backbone of the donor CPE effectively templates a planarized acceptor polymer, leading to excited states that are highly delocalized along the polymer backbone.

Assessment of volatile compound profiles and the deduced sensory significance of virgin olive oils from the progeny of Picual×Arbequina cultivars.

Volatile compounds are responsible for most of the sensory qualities of virgin olive oil and they are synthesized when enzymes and substrates come together as olive fruit is crushed during the industrial process to obtain the oil. Here we have studied the variability among the major volatile compounds in virgin olive oil prepared from the progeny of a cross of Picual and Arbequina olive cultivars (Olea europaea L.). The volatile compounds were isolated by SPME, and analyzed by HRGC-MS and HRGC-FID. Most of the volatile compounds found in the progeny's oil are produced by the enzymes in the so-called lipoxygenase pathway, and they may be clustered into different groups according to their chain length and polyunsaturated fatty acid origin (linoleic and linolenic acids). In addition, a group of compounds derived from amino acid metabolism and two terpenes also contributed significantly to the volatile fraction, some of which had significant odor values in most of the genotypes evaluated. The volatile compound content of the progeny was very varied, widely transgressing the progenitor levels, suggesting that in breeding programs it might be more effective to consider a larger number of individuals within the same cross than using different crosses with fewer individuals. Multivariate analysis allowed genotypes with particularly interesting volatile compositions to be identified and their flavor quality deduced.

Modulating oxidoreductase activity modifies the phenolic content of virgin olive oil.

The effect of modifying polyphenol oxidase (PPO) and peroxidase (POX) activity during the extraction of virgin olive oil has been assessed in terms of its influence on the phenolic profile of the oil produced. These enzymes were modified by adding exogenous enzyme or specific inhibitors during the milling and subsequent kneading step, studying the effect on specific phenolic compounds in the oils. PPO is the main enzyme involved in phenolic oxidation at the milling step whereas POX activity seems to be the main influence during the kneading step. The data obtained suggest it is possible to increase the nutritional and organoleptic quality of virgin olive oil by inhibiting these enzymes during olive fruit processing. Treatment with the PPO inhibitor tropolone produced a twofold increase in the phenolic fraction, which would therefore seem to be an interesting strategy to improve the nutritional and organoleptic properties of virgin olive oil.

Variability of virgin olive oil phenolic compounds in a segregating progeny from a single cross in Olea europaea L. and sensory and nutritional quality implications.

Virgin olive oil phenolic compounds are responsible for its nutritional and sensory quality. The synthesis of phenolic compounds occurs when enzymes and substrates meet as olive fruit is crushed during the industrial process to obtain the oil. The genetic variability of the major phenolic compounds of virgin olive oil was studied in a progeny of the cross of Picual x Arbequina olive cultivars (Olea europaea L.). They belong to four different groups: compounds that included tyrosol or hydroxytyrosol in their molecules, lignans, flavonoids, and phenolic acids. Data of phenolics in the oils showed that the progeny displayed a large degree of variability, widely transgressing the genitor levels. This high variability can be of interest on breeding programs. Thus, multivariate analysis allowed to identify genotypes within the progeny particularly interesting in terms of phenolic composition and deduced organoleptic and nutritional quality. The present study has demonstrated that it is possible to obtain enough degree of variability with a single cross of olive cultivars for compounds related to the nutritional and organoleptic properties of virgin olive oil.

Epidemiology and clinical impact of infection in patients awaiting heart transplantation.

OBJECTIVES: The aim of this study was to determine the epidemiology and clinical impact of infections in patients awaiting heart transplantation. METHODS: We evaluated all patients considered for a heart transplant in our center over a period of 18 months over a period of 18 months from 2007 to 2009. The patients were followed up for 8 months or until death, transplant, or loss to follow-up. RESULTS: Ninety patients were included in the study. During follow-up, 25 infections were recorded in 22 heart transplant candidates (24.4%). Respiratory infections were the most frequent infection (12 bronchitis; 48.0%), followed by skin and soft tissue infections (four infections; 16.0%), intra-abdominal infections (four infectious diarrhea; 16.0%), bacteremia (three infections; 12.0%), and urinary tract infections (two infections; 2.0%). Age, comorbidity, sex, and diabetes were not found to be risk factors for infection. Twenty-four patients (26.7%) were transplanted during follow-up. Infection before transplantation was not associated with an increased risk of mortality or a higher rate of infection in the immediate post-transplant period. CONCLUSIONS: Infections are common in heart transplant candidates, affecting almost 25% of them. Respiratory tract infections are the most frequent type of infection. However, they are not associated with increased mortality in the immediate post-transplant period.

Deficient long-term response to pandemic vaccine results in an insufficient antibody response to seasonal influenza vaccination in solid organ transplant recipients.

BACKGROUND: Little is known about the long-term antibody response to the 2009-H1N1 vaccine in solid organ transplant recipients (SOTR) and its clinical repercussion on the efficacy of following 2010-2011 influenza vaccine. METHODS: We performed a multicenter prospective study in SOTR receiving one dose of the nonadjuvant 2010-2011 seasonal influenza vaccine and determined the immunological response at 5 weeks after vaccination. RESULTS: One hundred SOTR were included. Long-term antibody titers to the previous vaccine were only detected in one third of the patients. Patients with baseline titers had significantly higher seroprotection for the 2009-H1N1 strain (100% vs. 73%, relative risks [RR] 1.37, 95% confidence intervals [CI] 1.19-1.57; P=0.006), for H3N2 strain (100% vs. 62.2%, RR 1.61, 95% CI 1.36-1.90; P=0.005), and for B strain (100% vs. 69%; P=0.02). The seroconversion rate in patients with baseline titers was 90.9% vs. 73% (RR 2.97, 95% CI 0.75-11.74; P=0.07) for the 2009-H1N1 strain, 92.2% vs. 62.2% (RR 5.29, 95% CI 0.8-35.7; P=0.02) for the H3N2 strain, and 58.3% vs. 69% (P=0.45) for the B strain. CONCLUSIONS: SOTR response to the 2010-2011 influenza vaccine was not optimal. The response was related to baseline titers; however, most of the patients did not exhibit detectable antibodies at vaccination lacking long-term response. New strategies are necessary to improve vaccination efficacy.

Factors limiting the synthesis of virgin olive oil volatile esters.

The aim of the present work was to establish the limiting factors affecting the biosynthesis of volatile esters present in virgin olive oil (VOO). Oil volatile fractions of the main Spanish olive cultivars, Arbequina and Picual, were analyzed. It was observed that acetate esters were the most abundant class of volatile esters in the oils, in concordance with the high content of acetyl-CoA found in olive fruit, and that the content of C6 alcohols is limited for the synthesis of volatile esters during the production of VOO. Thus, the increase of C6 alcohol availability during VOO production produced a significant increase of the corresponding ester in the oils in both cultivars at two different maturity stages. However, the increase of acetyl-CoA availability had no effect on the VOO volatile fraction. The low synthesis of these C6 alcohols seems not to be due to a shortage of precursors or cofactors for alcohol dehydrogenase (ADH) activity because their increase during VOO production had no effect on the C6 alcohol levels. The experimental findings are compatible with a deactivation of ADH activity during olive oil production in the cultivars under study. In this sense, a strong inhibition of olive ADH activity by compounds present in the different tissues of olive fruit has been observed.

Precursor uptake assays and metabolic analyses in isolated tomato fruit chromoplasts.

BACKGROUND: Carotenoids are the most widespread group of pigments found in nature. In addition to their role in the physiology of the plant, carotenoids also have nutritional relevance as their incorporation in the human diet provides health benefits. In non-photosynthetic tissues, carotenoids are synthesized and stored in specialized plastids called chromoplasts. At present very little is known about the origin of the metabolic precursors and cofactors required to sustain the high rate of carotenoid biosynthesis in these plastids. Recent proteomic data have revealed a number of biochemical and metabolic processes potentially operating in fruit chromoplasts. However, considering that chloroplast to chromoplast differentiation is a very rapid process during fruit ripening, there is the possibility that some of the proteins identified in the proteomic analysis could represent remnants no longer having a functional role in chromoplasts. Therefore, experimental validation is necessary to prove whether these predicted processes are actually operative in chromoplasts. RESULTS: A method has been established for high-yield purification of tomato fruit chromoplasts suitable for metabolic studies. Radiolabeled precursors were efficiently incorporated and further metabolized in isolated chromoplast. Analysis of labeled lipophilic compounds has revealed that lipid biosynthesis is a very efficient process in chromoplasts, while the relatively low incorporation levels found in carotenoids suggest that lipid production may represent a competing pathway for carotenoid biosynthesis. Malate and pyruvate are efficiently converted into acetyl-CoA, in agreement with the active operation of the malic enzyme and the pyruvate dehydrogenase complex in the chromoplast. Our results have also shown that isolated chromoplasts can actively sustain anabolic processes without the exogenous supply of ATP, thus suggesting that these organelles may generate this energetic cofactor in an autonomous way. CONCLUSIONS: We have set up a method for high yield purification of intact tomato fruit chromoplasts suitable for precursor uptake assays and metabolic analyses. Using targeted radiolabeled precursors we have been able to unravel novel biochemical and metabolic aspects related with carotenoid and lipid biosynthesis in tomato fruit chromoplasts. The reported chromoplast system could represent a valuable platform to address the validation and characterization of functional processes predicted from recent transcriptomic and proteomic data.

Synthesis of volatile compounds of virgin olive oil is limited by the lipoxygenase activity load during the oil extraction process.

The aim of this work was to determine whether the lipoxygenase (LOX) activity is a limiting factor for the biosynthesis of virgin olive oil (VOO) volatile compounds during the oil extraction process. For this purpose, LOX activity load was modified during this process using exogenous LOX activity and specific LOX inhibitors on olive cultivars producing oils with different volatile profiles (Arbequina and Picual). Experimental data suggest that LOX activity is a limiting factor for the synthesis of the oil volatile fraction, this limitation being significantly higher in Picual cultivar than in Arbequina, in line with the lowest content of volatile compounds in the oils obtained from the former. Moreover, there is evidence that this limitation of LOX activity takes place mostly during the milling step in the process of olive oil extraction.

The effect of challenge method on methacholine-induced changes in sensitivity and air trapping.

BACKGROUND: The methacholine challenge test performed with the tidal breathing method induces a greater fall in FEV(1) than the dosimeter method; however, the effect of the challenge method on methacholine-induced fall in FVC has not been investigated. OBJECTIVE: To determine the influence of the challenge method on methacholine-induced changes in FEV(1) and FVC. METHODS: Airway responsiveness to methacholine was determined by dosimeter method and tidal breathing method in 37 subjects with suspected asthma. The dosimeter was modified to deliver an identical volume to that obtained with the tidal breathing method and the same nebulizer model was used for the two challenges. The response was expressed by the provocative concentration of methacholine causing a 20% fall in FEV(1) (PC(20)) and by the percent fall in FVC at the PC(20) value relative to FVC after saline inhalation. RESULTS: The PC(20) values obtained with the tidal breathing method and the dosimeter method were similar, with geometric mean values of 3.15 (95%CI, 1.85-5.34 mg/mL) and 2.51 (1.37-4.61 mg/mL, P = 0.092), respectively. The percent fall in FVC at the PC(20) value obtained with the dosimeter was significantly greater than that obtained with the tidal breathing method, with mean values of 11.8 (95%CI, 10.0-13.5%) and 9.4 (95%CI, 8.1-10.8, P = 0.002), respectively. CONCLUSIONS: Differences in methacholine PC(20) values obtained with the two challenge methods recommended in guidelines may be overcome by introducing some technical modifications in the dosimeter method. However, the technical factors that affect methacholine sensitivity and air trapping are at least partially different.