IDENTIFICATION OF BACTERIOCIN-PRODUCING LACTIC ACID BACTERIA AND IN VITRO EVALUATION OF THEIR POTENTIAL ROLE IN MASTITIS CONTROL

Bovine mastitis is one of the most significant causes of economic losses for the dairy industry. On the other hand, public health authorities advise prudent use of antibiotics because they could promote bacterial resistance and leave residues in food chain. The dairy industry could benefit from the development of safe antimicrobial agents and bacteriocins could be attractive alternatives to antibiotics. Due to the safety of Lactic acid bacteria (LAB), their bacteriocins have the potential to be used as antimicrobials in veterinary clinical application. We analyzed the efficacy of antibacterial substances produced by bacteriocinogenic Lactococcus lactis subsp. lactis strains against contagious and environmental mastitis pathogens. Thereafter, we investigated how lactococcal strains or their bacteriocins could influence mammary gland innate immune response in vitro. Out of 65 LAB strains tested, 3 were active against mastitis pathogens: 2 strains produced Nisin, one Lacticin 481 and in addition a novel molecule with likely antibacterial activity. To analyze the immune response of mammary epithelial cells when stimulated with lactococcal strains or bacteriocins, a stabilized epithelial cell line, BME-UV1, was used. Both lactococcal live cultures and their antibacterial products were shown to modulate the non-specific immune response of BME-UV1 cells: Lysozyme and N-acetil-\u3b2-D-glucosaminidase excretion were overall enhanced by bacteriocins and live-culture treatments, while intracellular amounts were unaffected by treatments. Proinflammatory cytokine expression of treated BME-UV1 was similar to that observed in control cells, except for Lactococcus lactis subsp. lactis SL153. Such strain induced a significant reduction of TNF\u3b1 transcriptional level. The stimulation of enzyme secretion due to the administration of lactococci or of their antibacterial products, with potential enhancement of pathogens cleaning, can be of interest for the prevention of intra mammary infections. In addition, Lactococcus lactis subsp. lactis SL153 strain could be advantageous for its potential anti-inflammatory properties and could be of interest for the development of intra-mammary probiotic treatments

Causal inference concepts applied to three observational studies in the context of vaccine development : from theory to practice

Background Randomized controlled trials are considered the gold standard to evaluate causal associations, whereas assessing causality in observational studies is challenging. Methods We applied Hill's Criteria, counterfactual reasoning, and causal diagrams to evaluate a potentially causal relationship between an exposure and outcome in three published observational studies: a) one burden of disease cohort study to determine the association between type 2 diabetes and herpes zoster, b) one post-authorization safety cohort study to assess the effect of AS04-HPV-16/18 vaccine on the risk of autoimmune diseases, and c) one matched case-control study to evaluate the effectiveness of a rotavirus vaccine in preventing hospitalization for rotavirus gastroenteritis. Results Among the 9 Hill's criteria, 8 (Strength, Consistency, Specificity, Temporality, Plausibility, Coherence, Analogy, Experiment) were considered as met for study c, 3 (Temporality, Plausibility, Coherence) for study a, and 2 (Temporary, Plausibility) for study b. For counterfactual reasoning criteria, exchangeability, the most critical assumption, could not be tested. Using these tools, we concluded that causality was very unlikely in study b, unlikely in study a, and very likely in study c. Directed acyclic graphs provided complementary visual structures that identified confounding bias and helped determine the most accurate design and analysis to assess causality. Conclusions Based on our assessment we found causal Hill's criteria and counterfactual thinking valuable in determining some level of certainty about causality in observational studies. Application of causal inference frameworks should be considered in designing and interpreting observational studies

Combinatorial Discriminant Analysis Applied to RNAseq Data Reveals a Set of 10 Transcripts as Signatures of Exposure of Cattle to Mycobacterium avium subsp. paratuberculosis

Paratuberculosis or Johne's disease in cattle is a chronic granulomatous gastroenteritis caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP). Paratuberculosis is not treatable; therefore, the early identification and isolation of infected animals is a key point to reduce its incidence. In this paper, we analyse RNAseq experimental data of 5 ELISA-negative cattle exposed to MAP in a positive herd, compared to 5 negative-unexposed controls. The purpose was to find a small set of differentially expressed genes able to discriminate between exposed animals in a preclinical phase from non-exposed controls. Our results identified 10 transcripts that differentiate between ELISA-negative, clinically healthy, and exposed animals belonging to paratuberculosis-positive herds and negative-unexposed animals. Of the 10 transcripts, five (TRPV4, RIC8B, IL5RA, ERF, CDC40) showed significant differential expression between the three groups while the remaining 5 (RDM1, EPHX1, STAU1, TLE1, ASB8) did not show a significant difference in at least one of the pairwise comparisons. When tested in a larger cohort, these findings may contribute to the development of a new diagnostic test for paratuberculosis based on a gene expression signature. Such a diagnostic tool could allow early interventions to reduce the risk of the infection spreading

Combinatorial Discriminant Analysis Applied to RNAseq Data Reveals a Set of 10 Transcripts as Signatures of Exposure of Cattle to Mycobacterium avium subsp. paratuberculosis

Paratuberculosis or Johne's disease in cattle is a chronic granulomatous gastroenteritis caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP). Paratuberculosis is not treatable; therefore, the early identification and isolation of infected animals is a key point to reduce its incidence. In this paper, we analyse RNAseq experimental data of 5 ELISA-negative cattle exposed to MAP in a positive herd, compared to 5 negative-unexposed controls. The purpose was to find a small set of differentially expressed genes able to discriminate between exposed animals in a preclinical phase from non-exposed controls. Our results identified 10 transcripts that differentiate between ELISA-negative, clinically healthy, and exposed animals belonging to paratuberculosis-positive herds and negative-unexposed animals. Of the 10 transcripts, five (TRPV4, RIC8B, IL5RA, ERF, CDC40) showed significant differential expression between the three groups while the remaining 5 (RDM1, EPHX1, STAU1, TLE1, ASB8) did not show a significant difference in at least one of the pairwise comparisons. When tested in a larger cohort, these findings may contribute to the development of a new diagnostic test for paratuberculosis based on a gene expression signature. Such a diagnostic tool could allow early interventions to reduce the risk of the infection spreading

Exploring the concept of functional vitamin D deficiency in pregnancy: impact of the interaction between 25-hydroxyvitamin D and parathyroid hormone on perinatal outcomes

Background: Associations of vitamin D with perinatal outcomes are inconsistent and few studies have considered the wider calcium metabolic system. Objectives: We aimed to explore functional vitamin D deficiency in pregnancy by investigating associations between vitamin D status, parathyroid hormone (PTH), and perinatal outcomes. Design: SCOPE (Screening for Pregnancy Endpoints) Ireland is a prospective cohort study of low-risk, nulliparous pregnant women. We measured serum 25-hydroxyvitamin D [25(OH)D] and PTH at 15 wk of gestation in 1754 participants. Results: Mean ± SD 25(OH)D was 56.6 ± 25.8 nmol/L (22.7 ± 10.3 ng/mL) and geometric mean (95% CI) PTH was 7.84 pg/mL (7.7, 8.0 pg/mL) [0.86 pmol/L (0.85, 0.88 pmol/L)]. PTH was elevated in 34.3% of women who had 25(OH)D <30 nmol/L and in 13.9% of those with 25(OH)D ≥75 nmol/L. Whereas 17% had 25(OH)D <30 nmol/L, 5.5% had functional vitamin D deficiency, defined as 25(OH)D <30 nmol/L with elevated PTH. Elevated mean arterial pressure (MAP), gestational hypertension, pre-eclampsia, and small-for-gestational-age (SGA) birth were confirmed in 9.2%, 11.9%, 3.8%, and 10.6% of participants, respectively. In fully adjusted regression models, neither low 25(OH)D nor elevated PTH alone increased the risk of any individual outcome. The prevalence of elevated MAP (19.1% compared with 9.7%) and SGA (16.0% compared with 6.7%) were highest (P < 0.05) in those with functional vitamin D deficiency compared with the reference group [25(OH)D ≥75 nmol/L and normal PTH]. The adjusted prevalence ratio (PR) and RR (95% CIs) for elevated MAP and SGA were 1.83 (1.02, 3.27) and 1.53 (0.80, 2.93), respectively. There was no effect of functional vitamin D deficiency on the risk of gestational hypertension (adjusted RR: 1.00; 95% CI: 0.60, 1.67) or pre-eclampsia (adjusted RR: 1.17; 95% CI: 0.32, 4.20). Conclusion: The concept of functional vitamin D deficiency, reflecting calcium metabolic stress, should be considered in studies of vitamin D in pregnancy. The SCOPE pregnancy cohort is registered at http://www.anzctr.org.au as ACTRN12607000551493

Determinazione del carvacrolo mediante HRGC-MS/FID nell'orata (Sparus aurata) di acquacoltura

Le proprietà antimicrobiche del carvacrolo sono state ampiamente descritte soprattutto sulla base di osservazioni in vitro. L’ampia diffusione “zootecnica” di Sparus aurata ha determinato in questi ultimi decenni il diffondersi di patologie batteriche come la pasteurellosi e la vibriosi. La sperimentazione è stata condotta per 45 giorni su orate (Sparus aurata) del peso medio di 150 g la cui razione alimentare giornaliera è stata incorporata con l’1% di carvacrolo, e su orate alimentate con il mangime base nello stesso periodo (gruppo controllo). La frazione lipidica è stata estratta dal muscolo mediante metodo di Folch modificato. L’identificazione del carvacrolo è stata effettuata via HRGC-MS. L’analisi quantitativa è stata valutata nelle stesse condizioni cromatografiche ma con detector a ionizzazione di fiamma (FID) in presenza di standard interno (n-dodecanolo). La procedura ha consentito il recupero del 98,97% circa del carvacrolo con un limite di rilevabilità pari a 0,5 ppm

A crisis of protection and safe passage: violence experienced by migrants/refugees travelling along the Western Balkan corridor to Northern Europe.

BACKGROUND: Pushed by ongoing conflicts and pulled by the desire for a better life, over one million migrants/refugees transited Balkan countries and arrived in Europe during 2015 and early 2016. To curb this influx, European countries instituted restrictive migration policies often characterized by building of razor-wire border fences and border closures. Among migrants/refugees who received mental health care in Serbia while travelling through Balkan countries to Northern Europe, we assessed the prevalence and patterns of violent events experienced including physical trauma. METHODS: A mixed methods study among migrants/refugees attending mobile mental health clinics run by Médecins sans Frontières (MSF) between July 2015 and June 2016, in Serbia - a main transit hub to European countries. Clinics were conducted according to MSF guidelines by experienced psychologists who were supported by cultural mediators. The main outcome measures were violent events and associated physical trauma. RESULTS: Of 992 migrants/refugees attending MSF mental health clinics, the majority (72%) were from Syria and Afghanistan and included vulnerable groups (14%) such as unaccompanied minors and pregnant women. The most frequent mental health symptoms/signs were anxiety (29%) and adjustment reactions (26%). Of the 992 migrants/refugees, 270 (27%) had experienced violent events during their journey. Signs of physical trauma due to acts of violence were seen in 223(22%) of the 992 individuals, 144 (65%) being perpetrated by State authorities and involving women (11%) and children (13%). Border closures along the Balkan route were associated with a dramatic decrease in registered migrants/refugee arrivals in Serbia. Conversely, among those that made it across the borders, an increasing linear trend in reported violent events was observed at MSF mental health clinics (X2 for linear trend, P <0 · 001). Qualitative evidence corroborated with quantitative findings. CONCLUSIONS: Nearly one-in-three migrants/refugees seen in MSF clinics experienced violent events including physical trauma along their journey. State authorities, including those in European countries were the perpetrators in over half of such events which were associated with border closures. There is "a crisis of protection and safe passage" which needs to change towards one of respect for the principles of international human rights and refugee law

A randomised controlled feasibility trial of intermittent theta burst stimulation with an open longer-term follow-up for young people with persistent anorexia nervosa (RaISE):Study protocol

OBJECTIVE: We present the protocol of a feasibility randomised controlled trial (RCT) of intermittent theta burst stimulation (iTBS) for young people with anorexia nervosa (AN). Effective first-line psychological therapies exist for young people with AN, but little is known about how to treat those who do not respond. Non-invasive neuromodulation, such as iTBS, could address unmet treatment needs by targeting neurocircuitry associated with the development and/or maintenance of AN.DESIGN: Sixty-six young people (aged 13-30 years) with persistent AN will be randomly allocated to receive 20 sessions of real or sham iTBS over the left dorsolateral prefrontal cortex in addition to their usual treatment. Outcomes will be measured at baseline, post-treatment (1-month post-randomisation) and 4-months post-randomisation (when unblinding will occur). Additional open follow-ups will be conducted at 12- and 24-months post-randomisation. The primary feasibility outcome is the proportion of participants retained in the study at 4-months. Secondary outcomes include AN symptomatology, other psychopathology, quality of life, service utilisation, neurocognitive processes, and neuroimaging measures.DISCUSSION: Findings will inform the development of a future large-scale RCT. They will also provide exploratory data on treatment efficacy, and neural and neurocognitive predictors and correlates of treatment response to iTBS in AN.</p

Antenatal vitamin D status is not associated with standard neurodevelopmental assessments at age 5 Years in a well-characterized prospective maternal-infant cohort

Background: Although animal studies show evidence for a role of vitamin D during brain development, data from human studies show conflicting signals. Objective: We aimed to explore associations between maternal and neonatal vitamin D status with childhood neurodevelopmental outcomes. Methods: Comprehensive clinical, demographic, and lifestyle data were collected prospectively in 734 maternal-infant dyads from the Cork BASELINE Birth Cohort Study. Serum 25-hydroxyvitamin D [25(OH)D] concentrations were quantified at 15 weeks of gestation and in umbilical cord sera at birth via a CDC-accredited liquid chromatography-tandem mass spectrometry method. Children were assessed at age 5 y through the use of the Kaufman Brief Intelligence Test (2nd Edition, KBIT-2) and the Child Behaviour Checklist (CBCL). Linear regression was used to explore associations between 25(OH)D and neurodevelopmental outcomes. Results: 25(OH)D concentrations were <30 nmol/L in 15% of maternal and 45% of umbilical cord sera and <50 nmol/L in 42% of mothers and 80% of cords. At age 5 y, the mean ± SD KBIT-2 intelligence quotient (IQ) composite score was 104.6 ± 8.6; scores were 107.2 ± 10.0 in verbal and 99.8 ± 8.8 in nonverbal tasks. Developmental delay (scores <85) was seen in <3% of children across all domains. The mean ± SD CBCL total problem score was 21.3 ± 17.5; scores in the abnormal/clinical range for internal, external, and total problem scales were present in 12%, 4%, and 6% of participants, respectively. KBIT-2 and CBCL subscale scores at 5 y were not different between children exposed to low antenatal vitamin D status, either at 30 or 50 nmol/L 25(OH)D thresholds. Neither maternal nor cord 25(OH)D (per 10 nmol/L) were associated with KBIT-2 IQ composite scores [adjusted β (95% CI): maternal –0.01 (−0.03, 0.02); cord 0.01 (−0.03, 0.04] or CBCL total problem scores [maternal 0.01 (−0.04, 0.05); cord 0.01 (−0.07, 0.09)]. Conclusion: In this well-characterized prospective maternal-infant cohort, we found no evidence that antenatal 25(OH)D concentrations are associated with neurodevelopmental outcomes at 5 y. The BASELINE Study was registered at www.clinicaltrials.gov as NCT01498965; the SCOPE Study was registered at http://www.anzctr.org.au as ACTRN1260700055149