The untargeted lipidomic profile of quarter milk from dairy cows with subclinical intramammary infection by non-aureus staphylococci.

This observational study determined the lipidome of cow milk during subclinical intramammary infection (IMI) by non-aureus staphylococci (NAS), also defined as coagulase-negative staphylococci, using an untargeted approach. Among the pathogens causing bovine IMI, NAS have become the most frequently isolated bacteria from milk samples. Although the application of system biology approaches to mastitis has provided pivotal information by investigating the transcriptome, proteome, peptidome, and metabolome, the milk lipidome during mammary gland inflammation remains undisclosed. To cover this gap, we determined the milk lipidome of 17 dairy cows with IMI caused by NAS (NAS-IMI), and we compared the results with those of healthy quarter milk from 11 cows. The lipidome was determined following a liquid chromatography-quadrupole time-of-flight mass spectrometry approach. Sixteen subclasses of lipids were identified in both groups of animals. From 2,556 measured lipids, the abundance of 597 changed more than 10-fold in quarter milk with NAS-IMI compared with healthy quarters. The results demonstrate the influence of NAS-IMI on the milk lipidome, implying significant changes in lipid species belonging to the family of triacylglycerols and sphingomyelins, and contribute to the understanding of inflammatory processes in the bovine udder, highlighting potential novel biomarkers for improving mastitis diagnostics.

Differential microRNA expression between decidual and peripheral blood natural killer cells in early pregnancy.

STUDY QUESTION: Have decidual natural killer (dNK) cells a different microRNA (miRNA or miR) expression pattern compared to NK cells circulating in the peripheral blood (pb) of healthy pregnant women in the first trimester of gestation? SUMMARY ANSWER: dNK cells have a unique miRNA profile, showing exclusive expression of a set of miRNAs and significant up- or down-regulation of most of the miRNAs shared with pbNK cells. WHAT IS KNOWN ALREADY: dNK cells differ from pbNK cells both phenotypically and functionally, and their origin is still debated. Many studies have indicated that miRNAs regulate several important aspects of NK cell biology, such as development, activation and effector functions. STUDY DESIGN, SIZE, DURATION: Decidua basalis and peripheral blood specimens were collected from women (n = 7) undergoing voluntary termination of gestation in the first trimester of pregnancy. dNK and pbNK cells were then highly purified by cell sorting. PARTICIPANTS/MATERIALS, SETTING, METHODS: miRNAs expression was analysed by quantitative RT-PCR (qRT-PCR)-based arrays using RNA purified from freshly isolated and highly purified pbNK and dNK cells. Results from arrays were validated by qRT-PCR assays. The bioinformatics tool ingenuity pathway analysis (IPA) was applied to determine the cellular network targeted by validated miRNAs and the correlated biological functions. MAIN RESULTS AND THE ROLE OF CHANCE: Herein, we identified the most differentially expressed miRNAs in NK cells isolated from peripheral blood and uterine decidua of pregnant women. We found that 36 miRNAs were expressed only in dNK cells and two miRNAs only in pbNK cells. Moreover, 48 miRNAs were commonly expressed by both NK cell preparations although at different levels: 28 were upregulated in dNK cells, while 15 were downregulated compared to pbNK cells. Validation of a selected set (n = 11) of these miRNAs confirmed the differential expression of nine miRNAs: miR-10b and miR-214 expressed only in dNK cells and miR-200a-3p expressed only in pbNK cells; miR-130b-3p, miR-125a-5p, miR-212-3p and miR-454 were upregulated while miR-210-3p and miR-132 were downregulated in dNK cells compared to pbNK cells. IPA network analysis identified a single network connecting all the miRNAs as well as their significant involvement in several classes of functions: 'Organismal injury, Reproductive system disease, Inflammatory disease' and 'Cellular development'. These miRNAs target molecules such as argonaute 2, tumour protein p53, insulin and other genes that belong to the same network and significantly influence cell differentiation and pregnancy. LIMITATIONS, REASONS FOR CAUTION: In the present study, the cellular network and biological functions modulated by miRNAs differentially expressed in dNK and pbNK cells were identified by IPA considering only molecules and relationships that were with confidence 'experimentally observed' in leucocytes. The decidual and pbNK cells that were analysed here are a heterogeneous population and further study will help to disentangle whether there are differences in miRNA production by the different subsets of NK cells. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study describing a different miRNA expression profile in dNK cells compared to matched pbNK cells during the first trimester of pregnancy. Our findings improved the body of knowledge on dNK cell biology and strongly suggest further investigation into the roles of miRNAs that are differentially expressed in human dNK compared to pbNK cells. Our results suggest that specific miRNAs can modulate dNK cell origin and functions, highlighting a potential role of this miRNA signature in human development and diseases. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by grants from the Istituto Pasteur, Fondazione Cenci Bolognetti, the European NoE EMBIC within FP6 (Contract number LSHN-CT-2004-512040), Istituto Italiano di Tecnologia, and Ministero dell'Istruzione, dell'Università e della Ricerca (Ricerche Universitarie), and from Università Politecnica delle Marche. There are no conflicts of interest to declare.

[Inhaled corticosteroids: Which regimens are appropriate?]. / Corticoïdes inhalés : quels schémas ?

Inhaled corticosteroids are the cornerstone of asthma management. Inhaled corticosteroid regimens differ slightly in various international guidelines on asthma management but are based on the principles of continuous treatment and titration to the lowest effective dose. Several recent studies, nevertheless, appear to demonstrate the potential value of preemptive or "pro re nata" regimens in infants and children. These studies were included in GINA 2015 for children 5 years of age and younger in whom discontinuous treatment is proposed as a second-line option. Should we change our practices after a critical reading of these studies?

Melatonin promotes Bax sequestration to mitochondria reducing cell susceptibility to apoptosis via the lipoxygenase metabolite 5-hydroxyeicosatetraenoic acid.

Extra-neurological functions of melatonin include control of the immune system and modulation of apoptosis. We previously showed that melatonin inhibits the intrinsic apoptotic pathway in leukocytes via stimulation of high affinity MT1/MT2 receptors, thereby promoting re-localization of the anti-apoptotic Bcl-2 protein to mitochondria. Here we show that Bcl-2 sequesters pro-apoptotic Bax into mitochondria in an inactive form after melatonin treatment, thus reducing cell propensity to apoptosis. Bax translocation and the anti-apoptotic effect of melatonin are strictly dependent on the presence of Bcl-2, and on the 5-lipoxygenase (5-LOX) metabolite 5-hydroxyeicosatetraenoic acid (5-HETE), which we have previously shown to be produced as a consequence of melatonin binding to its low affinity target calmodulin. Therefore, the anti-apoptotic effect of melatonin requires the simultaneous, independent interaction with high (MT1/MT2) and low (calmodulin) affinity targets, eliciting two independent signal transduction pathways converging into Bax sequestration and inactivation. MT1/MT2 vs. lipoxygenase pathways are activated by 10(-9) vs. 10(-5)M melatonin, respectively; the anti-apoptotic effect of melatonin is achieved at 10(-5)M, but drops to 10(-9)M upon addition of exogenous 5-HETE, revealing that lipoxygenase activation is the rate-limiting pathway. Therefore, in areas of inflammation with increased 5-HETE levels, physiological nanomolar concentrations of melatonin may suffice to maintain leukocyte viability.

[Acute cough in infants: impact of national guidelines on medical practices in a French department]. / Recommandations de l'Agence nationale de sécurité du médicament concernant le traitement de la toux aiguë du nourrisson : évaluation des pratiques médicales dans les Alpes-Maritimes.

CONTEXT AND AIM: Guidelines have been published regularly since 2010 by the ANSM (the Health Products Safety Agency) advising against antitussive drugs for infants because of their inefficacy and their side effects (convulsions, respiratory congestion). Antihistamines, mucolytics, and terpene-based suppositories have theoretically disappeared from infant pharmacopoeia. We assessed the degree of compliance with these guidelines on the part of health professionals. MATERIALS AND METHODS: From June 2012 to August 2012, 198 general practitioners and 44 pediatricians were assessed in the Alpes-Maritimes department of France by means of questionnaires. At the same time, an analysis of medical prescriptions was sought from the Regional Directorate of Medical Services (RDMS) of the PACA-Corse region Health Insurance. RESULTS: The rate of noncompliance with the contraindications was 14.9% (17.7% general practitioners and 2.3% pediatricians). The RDMS study showed a slight decrease in prescriptions for antitussives not recommended from 2011 to 2012: -21% for pediatricians, -15.5% for generalist physicians. CONCLUSION: Continued educational and informative campaigns are therefore needed to stop medical prescriptions of cough medicines for infants, which should also involve parents.

[Acute cough in infants: impact on families and pharmacists of contraindications of the Agence nationale de sécurité du médicament et des produits de santé (ANSM)]. / Recommandations de l'Agence nationale de sécurité du médicament et des produits de santé (ANSM) concernant la prise en charge de la toux aiguë du nourrisson : impact sur les familles et les pharmacies.

CONTEXT AND OBJECTIVE: Although commonplace and usually not serious, acute cough in the context of upper respiratory tract infection is a frequent reason for consultation and generates significant parental anxiety. Parents often request active drug intervention. Following the contraindications in infants of mucolytics, Hélicidine(®), antihistamines, and terpene-based suppositories, issued between 2010 and 2011 by the Agence nationale de sécurité du médicament et des produits de santé (ANSM), we wished to assess whether these contraindications were known by parents and applied in pharmacies. MATERIALS AND METHODS: An assessment of compliance with these contraindications was made by a double declarative prospective observational study in Nice, first with 29 pharmacists and pharmacy technicians (4-25 August 2012) and then with 289 parents of infants (December 2011 to April 2012). RESULTS: The rate of noncompliance with contraindications was 23.8 % for parents and 34.5 % of pharmacists and pharmacy technicians. Consumption of cough medicines was inversely correlated to the ability to perform a correct nasal wash (OR=2.3). Only 21 % of parents used nasal wash properly. Full-time work was a risk factor for noncompliance with contraindications (OR=1.91). CONCLUSION: ANSM contraindications still have a limited impact on pharmacists and families. Therefore, efforts must be pursued to stop delivering cough medicines for infants. The information and educational campaigns should also involve parents and help to improve nasal wash use.

[Impact on the indoor environment of allergic children of the medical counselor on indoor environment, after two successive visits at 6 months interval]. / Impact sur l'habitat de l'enfant atteint d'une affection respiratoire allergique du conseiller médical en environnement intérieur après deux visites successives à 6 mois d'intervalle.

BACKGROUND: The aim of this retrospective study was to assess the impact of a medical indoor environment counselor (MIEC) on the allergic child's indoor home environment, as well as the real-life experience of patients' families. METHODS: We enrolled 50 children (age, 4-18 years) with allergic respiratory illness (96 % asthmatics) from March 2011 to January 2012. During the first visit, the CMEI gave advice according to the results of the assessment. Home environmental exposures were assessed 6 months later. A satisfaction questionnaire was completed by the parents. RESULTS: We found a significant decrease in the presence of house dust mites (P = 0.0047), humidity, and molds (P = 0.0047) as well as volatile organic compounds (P = 0.0047). Smoking habits were not significantly changed (P = 0.083), nor was the presence of domestic pets (P = 0.3173). Over 74 % of the families were very satisfied with the CMEI's intervention. DISCUSSION: According to de Blay's study, a home visit by the MEIC increased compliance with mite reduction. The intervention to advise parents of asthmatic children on the risks of passive smoking was ineffective in reducing their children's exposure to environmental tobacco smoke. The advice given by the MEIC was better understood by the patients than that expressed by the medical teams. CONCLUSION: A targeted home-based environmental intervention increased the compliance to mite, humidity, and mold reduction. The role of the CMEI will undoubtedly develop: follow-up studies are necessary to justify their activity (cost-efficacy ratio of their intervention).

Prevalence of owner-reported behaviours in dogs separated from the litter at two different ages.

The present study examined the prevalence of behaviours in dogs separated from the litter for adoption at different ages. Seventy adult dogs separated from their dam and littermates and adopted between the ages of 30 and 40 days were compared with 70 adult dogs that had been taken from the litter for adoption at two months. Owners were asked to complete a questionnaire eliciting information on whether their dog exhibited potentially problematic behaviours when in its usual environment. Binary logistic regression analysis was performed to investigate whether the age at which the dog was separated from the litter might predispose it to developing undesirable behaviours. The odds of displaying destructiveness, excessive barking, fearfulness on walks, reactivity to noises, toy possessiveness, food possessiveness and attention-seeking were significantly greater for the dogs that had been removed from the litter earlier during the socialisation period. In addition, dogs purchased from a pet shop at 30 to 40 days of age were reported to exhibit some of the listed behaviours with a significantly higher frequency than dogs purchased from a pet shop at two months. No significant differences were observed with dogs obtained from other types of sources. The dogs in the youngest age group (18 to 36 months) had a higher probability of displaying destructiveness and tail chasing. These findings indicate that, compared with dogs that remained with their social group for 60 days, dogs that had been separated from the litter earlier were more likely to exhibit potentially problematic behaviours, especially if they came from a pet shop.

Benefit of the basophil activation test in deciding when to reintroduce cow's milk in allergic children.

BACKGROUND: Oral challenges are required to establish the persistence or resolution of IgE-mediated cow's milk allergy (CMA). Determining the appropriate timing for challenging is the main difficulty. The benefit of the basophil activation test (BAT) in predicting a child's reaction to the oral challenge was evaluated and compared to the specific IgE and skin prick tests' (SPT) results. METHODS: One hundred and twelve consecutive children with CMA admitted for an oral challenge to reassess their allergy were included. Allergen-induced basophil activation was detected as a CD63-upregulation by flow cytometry. RESULTS: Thirty-six children (32%) had a positive oral challenge. The percentage of activated basophils in patients with a positive challenge (mean = 20.9; SD = 18.8) was significantly higher than that of patients with a negative challenge (mean = 3.9; SD = 9.8, P < 0.0001), and was well correlated with the eliciting dose of cow's milk (P < 0.0001). The BAT had an efficiency of 90%, a sensitivity of 91%, a specificity of 90%, and positive and negative predictive values of 81% and 96% in detecting persistently allergic patients. The area under the ROC curve was 0.866. These scores were higher than those obtained with SPT and IgE values, whichever positivity cut-point was chosen. Referring to a decisional algorithm combining BAT, specific IgE and SPT allowed the correct identification of 94% of patients as tolerant or persistently allergic to cow's milk proteins (CMP) in our cohort. CONCLUSION: The BAT could be a valuable tool in the management of paediatric CMA in addition to specific IgE quantification and SPT, by contributing in determining whether an oral challenge can safely be undertaken.

Lipoxygenase-mediated pro-radical effect of melatonin via stimulation of arachidonic acid metabolism.

We have shown that melatonin immediately and transiently stimulates intracellular free radical production on a set of leukocytes, possibly as a consequence of calmodulin binding. We show here that melatonin-induced ROS are produced by lipoxygenase (LOX), since they are prevented by a set of LOX inhibitors, and are accompanied by increase of the 5-LOX product 5-HETE. LOX activation is accompanied by strong liberation of AA; inhibition of Ca(2+)-independent, but not Ca(2+)-dependent, phospholipase A2 (PLA2), prevents both melatonin-induced arachidonic acid and ROS production, whereas LOX inhibition only prevents ROS, indicating that PLA2 is upstream with respect to LOX, as occurs in many signaling pathways. Chlorpromazine, an inhibitor of melatonin-calmodulin interaction, inhibits both ROS and arachidonic acid production, thus possibly placing calmodulin at the origin of a melatonin-induced pro-radical pathway. Interestingly, it is known that Ca(2+)-independent PLA2 binds to calmodulin: our results are compatible with PLA2 being liberated by melatonin from a steady-state calmodulin sequestration, thus initiating an arachidonate signal transduction. These results delineate a novel molecular pathway through which melatonin may participate to the inflammatory response.

[Experience of extensive screening of Mycobacterium tuberculosis transmission to patients exposed to a pediatrician]. / Expérience du dépistage à large échelle de la transmission de Mycobacterium tuberculosis par un pédiatre libéral à ses patients.

OBJECTIVE: This study reports the investigation of the patients of a private-practice pediatrician with smear-positive tuberculosis. METHODS AND PATIENTS: One thousand six hundred and fifty-six children were screened. Two screenings (T0 and 3 months later) were proposed, with a tuberculin skin testing (TST) and a chest radiograph. A T-cell-based assay was performed on children with intermediary values. RESULTS: No active tuberculosis was identified. Skin tests on 1171 children (83.1%) were negative on screenings. Nearly all chest radiographs were normal (96.9% on the initial screening, 98.8% on the 2nd). T-cell-based assays were negative. Of the 803 children younger than 2 years of age, 583 (72.6%) were directed to prophylaxis, children older than 2 years of age were treated based on the 2nd screening as latent tuberculosis infection. Sixty non serious side effects were reported 54 children, most of were digestive. Prophylaxis was stopped in 52 cases, in 22 cases a side effect. Compliance to the 2 visits was good (87.7%). CONCLUSION: This investigation concerning a large number of children confirms limited transmission of Mycobacterium tuberculosis from a pediatrician with smear-positive tuberculosis to pediatric outpatients. Coordination screening by the tuberculosis control section is essential. The T-cell-based assay could better correlate the contamination risk to the intensity and the length of exposure compared with TST and could avoid screening a large number of patients.

[Impact of rapid influenza test during influenza epidemic in all febrile children less than 6 years old in a pediatric emergency department]. / Intérêt du test de diagnostic rapide de la grippe aux urgences pédiatriques chez tout enfant âgé de moins de 6 ans et fébrile, en période épidémique.

AIM: To determine the impact of rapid influenza test (RIT) on the prescription of additional tests, antibiotics and oseltamivir, and the influence of oseltamivir on clinical signs and parents' day work stoppage. METHODS: Prospective study in the pediatric emergency department of Nice University Hospital from 29th January 2007 to 3rd March 2007 including children from 1 month to 6 years old with fever greater or equal to 38.5 degrees C for less than 48 h. Virologic research on nasopharyngeal aspiration was: immunofluorescence, cell culture and RIT Quickvue. Clinical informations, additional tests and treatments were registered for each child. An antiviral treatment (oseltamivir) was proposed to children older than 1 year with positive RIT. Evolution at 7 days was evaluated by phone contact. RESULTS: One hundred and seventy-seven children were included (mean age 24 months, sex-ratio 1.88). The RIT was positive in 42.3% (n=75). Compared with cell culture, the sensibility, specificity, positive predictive value and negative predictive value of the RIT were, respectively, 95.6, 91.6, 88 and 97%. Clinical signs significantly correlated to influenza were: impairment, rhinitis and acute otitis media. In the RIT positive group, there were significantly less additional tests (13 versus 36) and particularly urinalysis (5 versus 19), and more spreading in the family (p=0.0002). There was not any significant difference concerning hospitalizations, antibiotic prescriptions, or parents' day work stoppage. CONCLUSION: During influenza epidemic, in a pediatric emergency department, RIT allows a reduction of additional tests in febrile young children, particularly urinalysis.