Macrosystematics of eutherian mammals combining HTS data to expand taxon coverage.

In the last few years high-throughput sequencing technologies have permitted significant advances in mammalian phylogenetic studies from a genomic perspective. However, these studies have been restricted to a sparse number of species with available reference genomes. Thus, several issues inside the eutherian mammals phylogeny remain unresolved. This may be due in part to limited taxon sampling, as taxonomic density is known to affect phylogenetic resolution. In this context, we present a protocol to increase taxon coverage using high-throughput sequencing data (RNA or DNA) generated for other biological studies and available in public databases. Following this procedure we addressed pending or controversial issues concerning the phylogenetic position of Dermoptera, Pholidota and Chiroptera, considering multiple and independent loci. Also for Chiroptera and Arctoidea we evaluated the relationships of the lineages that compose it. Although the maximum number of genes used is moderate (95), in some cases taxon coverage doubles that of previous related studies. Globally, all coalescent-based (STAR, MP-EST and ASTRAL) and concatenated (IQ-TREE and BEAST2) methods used for species tree reconstruction were consistent to each other and most of interrogated nodes received high statistical support.

Single-cell view of deep-sea microbial activity and intracommunity heterogeneity.

Microbial activity in the deep sea is cumulatively important for global elemental cycling yet is difficult to quantify and characterize due to low cell density and slow growth. Here, we investigated microbial activity off the California coast, 50-4000 m water depth, using sensitive single-cell measurements of stable-isotope uptake and nucleic acid sequencing. We observed the highest yet reported proportion of active cells in the bathypelagic (up to 78%) and calculated that deep-sea cells (200-4000 m) are responsible for up to 34% of total microbial biomass synthesis in the water column. More cells assimilated nitrogen derived from amino acids than ammonium, and at higher rates. Nitrogen was assimilated preferentially to carbon from amino acids in surface waters, while the reverse was true at depth. We introduce and apply the Gini coefficient, an established equality metric in economics, to quantify intracommunity heterogeneity in microbial anabolic activity. We found that heterogeneity increased with water depth, suggesting a minority of cells contribute disproportionately to total activity in the deep sea. This observation was supported by higher RNA/DNA ratios for low abundance taxa at depth. Intracommunity activity heterogeneity is a fundamental and rarely measured ecosystem parameter and may have implications for community function and resilience.

The composition of the lipid, protein and mineral fractions of quail breast meat obtained from wild and farmed specimens of Common quail (Coturnix coturnix) and farmed Japanese quail (Coturnix japonica domestica).

The present study was intended to answer 2 scientific hypotheses: 1) the quail species has a significant influence in quail breast meat composition; 2) the wild quail's meat presents healthier composition than their farmed counterparts. An analysis of the pectoral muscles of wild and captive common quails (Coturnix coturnix) and domestic quails (Coturnix japonica domestica) was performed. The content of fatty acids (FA), amino acids, total cholesterol, and vitamin E, some basic macro- and microminerals in the pectoral muscles of the 2 species of the genus Coturnix were analyzed. Regarding the quail species influence on meat composition, Japanese Quail (JQ) revealed better lipid composition, characterized by lower saturated FA (SFA; less 3.17 g/100 g of total fatty acids), higher polyunsaturated FA contents (PUFA; more 5.5 g/100 g of total fatty acids) and healthier polyunsaturated FA/saturated FA (P/S) and n-6/n-3 ratios and TI value (1.08, 9.54 and 0.60 vs. 0.76, 12.58, and 0.75, correspondingly). The absence of differences observed on amino acids partial sums and ratios reveals equality between species on protein nutritional quality. On the other hand, Common Quail (CQ) proved to be a better source of copper (0.181 mg/100 g of meat), iron (2.757 mg/100 g of meat), manganese (0.020 mg/100 g of meat), and zinc (0.093 mg/100 g of meat) than JQ. The comparison of farmed and wild specimens within CQ, showed that wild birds presented lower total cholesterol (less 8.32 mg/g of fresh meat) and total PUFA (less 4.26 g/100 g of total fatty acids), and higher n-3 PUFA contents (more 1.53 g/100 g of total fatty acids), which contributed to healthier P/S and n-6/n-3 ratios, but worst PI (1.60, 8.08, and 113.1 vs. 0.76, 12.58, and 100.8, respectively). The wild species revealed higher α-tocopherol content (2.40 vs. 1.49 µg/g of fresh meat. Differences observed on their mineral composition counterbalance each other. Under intensive production system and similar feeding and management conditions, the CQ develops better nutritional qualities than JQ. The comparison of wild and farmed species within CQ reveals more similarities than differences. Quail´s meat presents good nutritional quality and introduces variability to human's diet, which is much valued by consumers.

Stranded pellets in Fildes Peninsula (King George Island, Antarctica): New evidence of Southern Ocean connectivity.

Plastic and microplastic debris is transported by ocean currents over long distances, reaching remote areas, far from its original source. In Polar Regions, microplastics (MPs) can come from local activities or be transported from lower latitudes, with the former being the likely and major source. Although historically Antarctica was considered isolated from the global ocean, there is recent evidence of materials and organisms being transported in and out of the Southern Ocean, despite its multi-front structure. During the austral summer of 2019, beach surveys were conducted on the NW coast of the Fildes Peninsula (King George Island). The beach was characterised, and the first 2 cm of sediment from 5 quadrants (50 × 50 cm) along 100 m of the highest strandline were collected. Large microplastics (LMPs) and mesoplastics (MesoPs) were isolated, counted, measured, weighed and classified by shape. Polymer composition was analysed by FTIR and ageing estimated by Carbonyl Index. We found 293 items of LMPs (188 items) and MesoPs (105 items), with a total average density (±SD) of 234.4 ± 166 items m-2. Foams (130.4 ± 76.3), fragments (58.4 ± 56.0) and pellets (44.0 ± 50.5) were the most abundant shapes. The main polymers found were polystyrene, polypropylene, and polyethylene. We found pellets among the MesoPs, being the first record for beaches in Antarctica. The presence of these primary MPs south of 62°S not only alerts about their possible direct consequences on Antarctic ecosystems, but also gives empirical evidence for the passive entry of plastic debris from lower latitudes through cross-frontal exchanges, providing new evidence of a global connectivity of the Southern Ocean. Despite increasing research, knowledge of plastics dynamics and their impact in the Southern Ocean and Antarctica is still limited but certainly necessary.

[Staphylococcus aureus prostatic abscess and subdural empyema: a case report]. / Absceso prostático por Staphilococo aureus y empiema subdural: presentación de un caso.

INTRODUCTION AND OBJECTIVES: To report one case of prostatic abscess and subdural empyema by Staphylococcus aureus. METHODS: We describe the case of a 51 year old male patient who was diagnosed of prostatic abscess and subdural empyema by Staphilococcus aureus. We use clinical presentation and physical exploration based on rectal digital examination, as diagnostic approach method. And computerized axial tomography and transrectal ultrasonography, which allows the guided needle drainage of the abscess, as diagnostic confirmation methods. RESULTS: The clinical picture resolved with the transrectal ultrasonography guided needle aspiration of the abscess and conservative treatment with antibiotics and urinary diversion. CONCLUSIONS: Prostatic abscess is an uncommon entity nowadays. Provided the great variety of symptoms, a great degree of clinical suspicion is needed for the diagnosis, and once it is got it, immediate aggressive treatment must be initiated. Transrectal ultrasonography allows not only the diagnosis, but also the drainage of the abscess. The culture of the obtained material identifies the etiological agent and the most specific antibiotic therapy.

Evaluation of the bioaccessible gastric and intestinal fractions of heavy metals in contaminated soils by means of a simple bioaccessibility extraction test.

A study is made to evaluate the bioaccessibility of heavy metals in contaminated soils through a simple bioaccessibility extraction test (SBET), applied to the analysis of both the gastric and intestinal phases. Soils with high metal content of the Mapocho, Cachapoal, and Rancagua series were studied; they are located in suburban areas of large cities in the central valley of Chile. The bioaccessible concentrations of Cd, Cr, Cu, Ni, Pb, and Zn were related to the main physicochemical characteristics of the soils and to the chemical forms obtained by sequential extraction. The elements Cd, Cu, Ni, and Zn are distributed in the soils between the exchangeable fractions, bound to oxides, to organic matter, and in the residual fraction. On the other hand, Cr and Pb are found mainly in the fractions bound to organic matter and in the residual fraction. The three soils have a high Cu content, (640-2060 mg/kg), in the order Cachapoal > Rancagua > Mapocho. The SBET test allowed establishing a different bioaccessibility for the elements in the soil. Cu was notoriously bioaccessible in both the gastric and intestinal phases in the three soils, reaching more than 50% in the Cachapoal and Rancagua soils. The other elements, regardless of the soil, were bioaccessible only in one of the phases, more frequently in the gastric phase. The multiple correlation study indicates that the metal forms have a higher incidence than the soil's physicochemical factors on the extractability to evaluate the human oral bioaccessibility of the metals.

Microstructural characterization of degradation in a high-chromium alloy in the sugar industry.

A new high (24%)-chromium alloy was experimentally manufactured by combining the particular chemical composition of various materials with the use of thermal treatments to produce pump impellers for the sugar industry. The alloy metal was tested in situ in a sugar medium and then evaluated by electrochemical measurements; the two former procedures were associated with a nontraditional method to assess the susceptibility and resistance to corrosion. Potentiostatic and atomic absorption measurements were carried out to characterize the degradation and damage morphology by scanning electron microscopy (SEM) observations. The trials proved a good performance for the new alloy in the sugar medium in spite of nonsignificant localized damages involving erosion-corrosion mechanisms and surface microcrackings of the material.

[Advices for a better drug prescription]. / Conseils pour une meilleure prescription médicamenteuse.

The prescription of a pharmacological agent concludes most medical visits. The rational basis of drug prescription by the physician relies on numerous pharmacodynamic, pharmacokinetic, pragmatic and economic considerations. However, it is a pity to notice that one patient out of two does not strictly follow the pharmacological treatment as it was prescribed. This article analyses some problems of the patient-doctor relationship, which may reduce medication compliance, and provides practical advice for improving such a situation in order to enhance both efficacy and safety of drug prescribing.

The dopamine antagonist sch 23390 reverses dizocilpine-induced blockade of cocaine sensitization.

The present work examined the effects of pre-treatment with Sch 23390, a selective D(1) receptor antagonist, on the dizocilpine-induced blockade of sensitization to the locomotor-stimulating effect of cocaine. Rats were given either cocaine [15mgkg(-1)day(-1), intraperitoneally (i.p.)] from day 1 to day 5 (cocaine-experienced rats) or vehicle (cocaine-naïve rats). From day 6 to day 15, animals remained drug-free in their home cages. On day 16 rats received a challenge injection of cocaine (15mgkg(-1)) or vehicle, and were tested for sensitization to the locomotor-stimulating effect of cocaine. In cocaine-naïve rats the acute effect of cocaine was a 1.5 times increase in locomotor activity. In cocaine-experienced rats, the acute effects of cocaine were considerably more pronounced than in cocaine-naïve rats; the stimulating effect of cocaine in these animals was a doubling in locomotor activity. In cocaine-naïve rats, pre-treatment with dizocilpine (100microgkg(-1)), Sch 23390 (100microgkg(-1)) or a combination of the two drugs from day 1 to day 5 changed neither spontaneous locomotor activity nor cocaine stimulant activity. By contrast, cocaine-experienced animals that had been given 100microgkg(-1) dizocilpine from day 1 to day 5 failed to show the increase in locomotor activity when challenged with cocaine on day 16. Pre-treatment with Sch 23390 (100microgkg(-1)day(-1), i.p.) from day 1 to day 5 was found to prevent completely the cocaine anti-sensitization properties of 100microgkg(-1) dizocilpine, but failed to prevent cocaine sensitization. On the other hand, horizontal activity in cocaine-experienced rats that had been given dizocilpine (100microgkg(-1)) 15min before cocaine challenge on day 16 was higher than in corresponding controls. It is concluded that prevention of cocaine sensitization by dizocilpine may be related to the events set into motion by the NMDA antagonist at the level of dopaminergic transmission involving D(1) receptors.

Anticonvulsant and sodium channel-blocking properties of novel 10,11-dihydro-5H-dibenz[b,f]azepine-5-carboxamide derivatives.

A series of esters of the major metabolite of oxcarbazepine (2), 10, 11-dihydro-10-hydroxy-5H-dibenz[b,f]azepine-5-carboxamide, were synthesized and evaluated for their anticonvulsant and brain sodium channel-blocking properties. The compounds were assayed intraperitoneally and per os in rats against seizures induced by maximal electroshock (MES). Neurologic deficit was evaluated by the rotarod test. The enantiomeric acetates (R)-11 and (S)-12 were the most active of the series against MES-induced seizures with oral ED(50) values at t(max) of 10.9 +/- 2.3 and 4.7 +/- 0.9 mg/kg, respectively. After intraperitoneal administration, carbamazepine (1) behaved more potently than 2 and all other new dibenz[b, f]azepine-5-carboxamide derivatives in the MES test; compounds 2 and 12 were equally potent. In the rotarod test, low doses of 1 produced considerable motor impairment, which did not occur with 2, enantiomeric alcohols (S)-6, (R)-7, and racemic alcohol 8, or racemic acetate 10 or (R)-11. The potencies of the racemic and enantiomerically pure alcohols 8, (S)-6, and (R)-7 derived from 2 in the MES and rotarod test were found to be similar between them, and consequently they exhibit similar protective index values. All three forms of the alcohol and their corresponding acetates (pairs 8 & 10, 6 & 12, and 7 & 11) were found to differ in the MES or rotarod tests; the ED(50) value for (S)-6 against MES-induced seizures was nearly 3-fold that for (S)-12. The protective index also differed markedly between all stereoisomers of the alcohol and their corresponding acetates, most pronouncedly for compound (S)-12 which attained the highest value (12.5) among all compounds tested. Blockade of voltage-sensitive sodium channels was studied by investigating [(3)H]batrachotoxinin A 20-alpha-benzoate ([(3)H]BTX) binding. Acetates (R)-11 and (S)-12 were more potent than the standards 1 and 2 at inhibiting the binding of [(3)H]BTX to sodium channels and the influx of (22)Na(+) into rat brain synaptosomes. It is concluded that acetates (R)-11 and (S)-12 are not simple metabolic precursors of alcohols (R)-7 and (S)-6 in rodents but that they possess anticonvulsant and sodium channel-blocking properties in their own right.

The O-methylated derivative of L-DOPA, 3-O-methyl-L-DOPA, fails to inhibit neuronal and non-neuronal aromatic L-amino acid decarboxylase.

The present study examined whether the O-methylated derivative of L-DOPA, 3-O-methyl-L-DOPA (3-OM-L-DOPA), inhibits neuronal (brain) and non-neuronal (liver and kidney) aromatic L-amino acid decarboxylase (AADC) activity. The incubation of brain, liver and kidney homogenates with 3-OM-L-DOPA (5 mM) did not result in the formation of 3-methoxytyramine, the compound expected to result from the decarboxylation of 3-OM-L-DOPA. Incubation of tissue homogenates with L-DOPA resulted in a concentration-dependent formation of dopamine, revealing K(m) values (in mM) of similar magnitude for brain (0.8), liver (1.6) and kidney (1.0). Both benserazide and L-5-hydroxytryptophan (L-5-HTP) were found to produce concentration dependent decreases in AADC activity with K(i) values in the microM range. By contrast, 3-OM-L95% reduction) in liver and kidney AADC activity accompanied by a marked decrease (49% reduction) in brain AADC activity. By contrast, the administration of 30 mg/kg (p.o.) 3-OM-L-DOPA, which generates levels in brain, liver and kidney six-fold those in L-DOPA-treated rats, was found to change neither neuronal nor non-neuronal AADC activity. In conclusion, 3-OM-L-DOPA fails to interact with neuronal and non-neuronal AADC, either as substrate or inhibitor.

BIA 3-202, a novel catechol-O-methyltransferase inhibitor, enhances the availability of L-DOPA to the brain and reduces its O-methylation.

1-[3,4-Dihydroxy-5-nitrophenyl]-2-phenyl-ethanone (BIA 3-202) is a new long-acting catechol-O-methyltransferase (COMT) inhibitor with limited access to the brain. The present study evaluated the interference of BIA 3-202 upon levels of L-3,4-dihydroxyphenylalanine (L-DOPA) and metabolites in plasma (3-O-methyl-L-DOPA) and brain [3-O-methyl-L-DOPA, dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA)] in rats orally treated with L-DOPA (20 mg/kg) plus benserazide (30 mg/kg). At different time points (1, 3 and 6 h) after the administration of BIA 3-202 (0, 3, 10 and 30 mg/kg) or L-DOPA plus benserazide, rats were sacrificed and the right striatum was quickly dissected out and stored for the assay of L-DOPA, 3-O-methyl-L-DOPA, dopamine and amine metabolites. Levels of L-DOPA, 3-O-methyl-L-DOPA, dopamine, DOPAC and HVA in the striatum in L-DOPA plus benserazide-treated rats were higher than in vehicle-treated rats. However, this increase in striatal L-DOPA, dopamine, DOPAC and HVA was, in a dose- and time-dependent manner, even higher (P<0.05) in rats given BIA 3-202 (3, 10 and 30 mg/kg). This effect was accompanied by a marked decrease in 3-O-methyl-L-DOPA levels in the striatum of L-DOPA plus benserazide-treated rats. Increases in levels of L-DOPA and decreases in 3-O-methyl-L-DOPA levels in plasma also accompanied the administration of BIA 3-202. BIA 3-202 did not significantly affect levels of DOPAC and HVA in the striatum in vehicle-treated rats. It is concluded that administration of BIA 3-202 enhances the availability of L-DOPA to the brain by reducing its O-methylation in the periphery, which may prove beneficial in parkinsonian patients treated with L-DOPA plus an aromatic amino acid decarboxylase inhibitor.

Metabolism of two new antiepileptic drugs and their principal metabolites S(+)- and R(-)-10,11-dihydro-10-hydroxy carbamazepine.

BIA 2-093 and BIA 2-059 are two stereoisomers under development as new antiepileptic drugs. They act as prodrugs for the corresponding hydroxy derivatives (S(+)- or R(-)-10,11-dihydro-10-hydroxy carbamazepine, respectively) which are known to be the active metabolites of the antiepileptic drug oxcarbazepine (OXC). The purpose of this study was to define the metabolic pathway especially in terms of stereoselectivity, and to estimate the possibility of racemization in humans. For in vivo studies, the rat, mouse and rabbit were chosen as models in order to cover a broad spectrum of metabolic activity. In addition, incubations with liver microsomes from these three species plus dog and monkey were compared to results obtained with human liver microsomes. It was found that both drugs were almost instantly hydrolysed to the corresponding 10-hydroxy compounds in mice, rats and rabbits. Mice and rabbits were not able to oxidize the 10-hydroxy compounds to OXC in significant amounts. In the rat, BIA 2-093 also gave origin to OXC, whereas BIA 2-059 resulted in the formation of OXC and the trans-diol metabolite in equal amounts. It could be shown that the rat is able to reduce the formed OXC in liver to S(+)-10-hydroxy metabolite, resulting in a loss of enantiomeric purity after treatment with BIA 2-059 rather than in the case of BIA 2-093. Human liver microsomes hydrolysed BIA 2-093 and BIA 2-059 to their corresponding 10-hydroxy compounds and to OXC in a very small extent with BIA 2-093 only. Therefore, BIA 2-093 and BIA 2-059 seem to be preferable drugs over OXC since they most likely exhibit a 'cleaner' metabolism. From a therapeutic point of view BIA 2-059 would be less appropriate than BIA 2-093 for the purpose of treating epileptic patients due to its propensity to undergo inactivation to the trans-diol.

Synthesis, anticonvulsant properties and pharmacokinetic profile of novel 10,11-dihydro-10-oxo-5H-dibenz/b,f/azepine-5-carboxamide derivatives.

A series of novel derivatives of oxcarbazepine (5), 10,11-dihydro-10-oxo-5H-dibenz/b,f/azepine-5-carboxamide was synthesised and evaluated for their anticonvulsant activity and sodium channel blocking properties. The oxime 8 was found to be the most active compound from this series, displaying greater potency than its geometric isomer 9 and exhibiting also the highest protective index value. Importantly, the metabolic profile of 8 differs from the already established dibenz/b,f/azepine-5-carboxamide drugs such as 1 and 5 which undergo rapid and complete conversion in vivo to several biologically active metabolites. In contrast 8 is metabolised to only a very minor extent leading to the conclusion that the observed anti-convulsant effect is solely attributable to 8. It is concluded that 8 may be as effective as 1 and 5 at controlling seizures and that the low toxicity and consequently high protective index should provide the compound with an improved side-effect profile.

Toxicity of a GM3 cancer vaccine in Macaca fascicularis monkey: a 12-month study.

GM3 is a ganglioside that has been biochemically identified as dominating the cell surface of several human tumours, but is also found on human normal cells at much lower density. Since GM3 is widely distributed in essentially all types of animal cells, there is a conflict with the concepts of tumour-associated antigen, immunogen, and toxicity. We have designed a GM3-based cancer vaccine for the treatment of human breast and melanoma tumours. Prior to the Phase I clinical trial, we carried out a 12-month dose repeated toxicity study in five male Macaca fascicularis monkeys. Four male monkeys were treated with placebo in a similar way. During the study, no differences were observed between control and treated monkeys related to daily clinical observations (other than local damage) including rectal temperature, blood pressure, respiratory and cardiac rates, weight gain, biochemical and hematological parameters (with the exception of transitory pathological changes), and anti-DNA and anti-nuclear antibodies, although treated monkeys consistently developed both IgM- and IgG-specific anti-GM3 antibodies. Sixty per cent of treated monkeys developed moderate local reactions at the injection site, which disappeared without sequels. We concluded that this GM3 cancer vaccine overcame in monkeys the natural tolerance to GM3 ganglioside evidenced by a strong immune response, while the local reactions elicited-were transitory without apparent important systemic toxicity effects.

Endoscopic papillotomy under ultra-sonographic control.

A new endoscopic method for performing endoscopic sphincterotomy is described. Only after correct positioning of the papillotomy device has been ascertained can papillotomy be carried out. We have used ultra-sonographic examination to confirm the correct placement of the papillotomy device. This technique was first employed on a pregnant patient in 6-13-1988. After that we performed this procedure in another twelve patients. We believe it is an easy procedure and perhaps may be used for all patients with common bile duct stones of less than 1.2 cm of diameter without other bilio-pancreatic pathological condition.

[Levosimendan reduces mortality in cardiac surgery: a systematic review and meta-analysis]. / Levosimendán disminuye la mortalidad en cirugía cardíaca: revisión sistemática y metaanálisis.

OBJECTIVES: To evaluate whether Levosimendan has any impact on peri-operative mortality in cardiac surgery. MATERIAL AND METHODS: A literature search was performed with the main purpose of looking for mortality in cardiac surgery. Data tabulated by both the fixed effects model and the random effects model were analysed based on the method by Peto. The statistical heterogeneity and inconsistency was estimated and quantified, as well as performing a sensitivity analysis to test the influence of individual studies on the overall result. All the calculations and statistical analyses were performed using the software; "Comprehensive Meta-Analysis" V2.0 and "StatsDirect statistical software" V 2.7.8. At the same time, possible publication bias was also determined. RESULTS: A total of 169 articles were found, from which 13 studies were finally included in the meta-analysis; 12 randomised and one of them retrospective with paired data. The global analysis showed that the use of levosimendan was associated with a significant reduction in postoperative mortality. CONCLUSIONS: The meta-analysis showed that the use of Levosimendan was associated with a reduction in mortality in patients subjected to cardiac surgery, although there is still no definitively clear evidence. Additional randomised and multicentre clinical studies, with a much larger number of patients are required, in which hospital mortality and mortality at 6 months are analysed as the primary outcomes.

Profiling allergic asthma volatile metabolic patterns using a headspace-solid phase microextraction/gas chromatography based methodology.

Allergic asthma represents an important public health issue with significant growth over the years, especially in the paediatric population. Exhaled breath is a non-invasive, easily performed and rapid method for obtaining samples from the lower respiratory tract. In the present manuscript, the metabolic volatile profiles of allergic asthma and control children were evaluated by headspace solid-phase microextraction combined with gas chromatography-quadrupole mass spectrometry (HS-SPME/GC-qMS). The lack of studies in breath of allergic asthmatic children by HS-SPME led to the development of an experimental design to optimize SPME parameters. To fulfil this objective, three important HS-SPME experimental parameters that influence the extraction efficiency, namely fibre coating, temperature and time extractions were considered. The selected conditions that promoted higher extraction efficiency corresponding to the higher GC peak areas and number of compounds were: DVB/CAR/PDMS coating fibre, 22 °C and 60 min as the extraction temperature and time, respectively. The suitability of two containers, 1L Tedlar® bags and BIOVOC®, for breath collection and intra-individual variability were also investigated. The developed methodology was then applied to the analysis of children exhaled breath with allergic asthma (35), from which 13 had also allergic rhinitis, and healthy control children (15), allowing to identify 44 volatiles distributed over the chemical families of alkanes (linear and ramified) ketones, aromatic hydrocarbons, aldehydes, acids, among others. Multivariate studies were performed by Partial Least Squares-Discriminant Analysis (PLS-DA) using a set of 28 selected metabolites and discrimination between allergic asthma and control children was attained with a classification rate of 88%. The allergic asthma paediatric population was characterized mainly by the compounds linked to oxidative stress, such as alkanes and aldehydes. Furthermore, more detailed information was achieved combining the volatile metabolic data, suggested by PLS-DA model, and clinical data.