Effect of rumen fluke on feed digestibility and methane emissions in sheep.

Thirty-six ewes aged 18 months were assigned to one of three groups (n = 12) on the basis of faecal egg count (FEC) for rumen fluke; C: control (FEC = 0-5 epg), T: affected (FEC ≥ 6 epg) and treated, NT: affected (FEC ≥ 6 epg) and not treated. T ewes were treated with oxyclozanide 14 days prior to the trial commencing. Ewes were fed grass silage ad libitum. Digestibility crates and respiration chambers were used to measure feed digestibility and methane production. Dry matter intake (DMI), feed digestibility and CH4/DMI did not differ (P > 0.05) between treatments. However, CH4/live weight (LW) was significantly greater (P < 0.001) for NT ewes, while that of C and T ewes were similar. This study indicates that a high rumen fluke burden can increase methane emissions but only when expressed on a body weight basis (per kg LW). As the link between rumen fluke infection and methane production has not previously been investigated within the literature, these findings demonstrate the need for further research to better understand these relationships.

Comparison of polysaccharide glycoconjugates as candidate vaccines to combat Clostridiodes (Clostridium) difficile.

Two known Clostridiodes (Clostridium) difficile surface antigens, a lipoteichoic acid (LTA) and a polysaccharide (PS-II) were isolated and purified in order to prepare glycoconjugate vaccines to the carrier protein human serum albumin utilising a reductive amination strategy. Mice and rabbits were immunized with a prime and two boost strategy and the resulting sera were examined for their ability to recognise the purified homologous antigens and subsequently killed whole cells of C. difficile strains and other Clostridia species. Immunisation derived antisera from rabbits and mice, recognised all strains of C. difficile vegetative cells examined, with generally similar titers from animals that received the LTA or the PS-II conjugates. Sera raised to the LTA conjugates were able to recognise other Clostridia species C. butyricum, C. bifermentans and C. subterminale whereas sera raised to the PS-II conjugates were not. These LTA and PS-II sera recognised live cells in an immunofluorescence assay and were also able to recognise the spore form of the bacterium. This study has confirmed that the LTA and PS-II polysaccharides are both highly conserved surface polymers of C. difficile that are easily accessible to the immune system and as such may have potential as vaccine antigens or as targets for therapeutics to combat C. difficile infection.

[Cervicofacial nontuberculous mycobacteria in children: Clinical, microbiological and therapeutic features. A retrospective study and literature review]. / Adénites cervico-faciales à mycobactéries atypiques de l'enfant : caractéristiques cliniques, microbiologiques et thérapeutiques. Étude rétrospective et revue de la littérature.

BACKGROUND: Nontuberculous mycobacteria (NTM) is a common cause of lymphadenitis. A rise in incidence has been reported. Our main aim was to describe the clinical features, microbiological aspects and treatment of the disease. METHODS: We conducted a retrospective, monocentric study between January 2008 and December 2017 (University Hospital of Nantes). INCLUSION CRITERIA: age<18 years, 1 positive lymph node specimen with identification of the species in culture, head-and-neck localization. RESULTS: Forty-nine patients were enrolled from 2008 to 2017. Median age was 28 months (range: 6-141 months). Median time to confirmation of diagnosis was 2.1 months (range: 0.7-6 months). The sites encountered were mandibular (45%), cervical (33%), and parotid (16%). The main clinical signs were a tender nodule (70%), purplish nodule (59%) or painless nodule (83%), without fever (88%). The species identified were: Mycobacterium avium (n=26), M. lentiflavum (n=13), M. intracellulare (n=7), M. malmoense (n=2) and M. scrofulaceum (n=1). Antibiotic treatment was frequent (77% of cases). DISCUSSION: This study is the second largest French cohort of NTM lymphadenitis in children with microbiological confirmation. The most frequent presentation was a tender, purplish, and painless mandibular nodule. The predominant species was M. avium. M. lentiflavum, which emerged during our study, does not figure in any European studies before 2014 but appears in the most recent studies. The effects of discontinuation of mandatory BCG immunization in France in NMT is not statistically demonstrable here due to lack of relevant data prior to 2007. CONCLUSION: A possible diagnosis of NTM lymphadenitis should not be overlooked in children presenting painless, purplish, cervicofacial tumefaction.

Anderson Mobility Gap Probed by Dynamic Coherent Backscattering.

We use dynamic coherent backscattering to study one of the Anderson mobility gaps in the vibrational spectrum of strongly disordered three-dimensional mesoglasses. Comparison of experimental results with the self-consistent theory of localization allows us to estimate the localization (correlation) length as a function of frequency in a wide spectral range covering bands of diffuse transport and a mobility gap delimited by two mobility edges. The results are corroborated by transmission measurements on one of our samples.

Ruling out false-positive urinary Legionella pneumophila serogroup 1 and Streptococcus pneumoniae antigen test results by heating urine.

We report here false-positive urinary Legionella pneumophila serogroup 1 and Streptococcus pneumoniae antigen test results due to rabbit antilymphocyte serum treatment and provide a simple and fast solution to rule them out by heating urine.

Recurrent scattering and memory effect at the Anderson localization transition.

We report on ultrasonic measurements of the propagation operator in a strongly scattering mesoglass. The backscattered field is shown to display a deterministic spatial coherence due to a remarkably large memory effect induced by long recurrent trajectories. Investigation of the recurrent scattering contribution directly yields the probability for a wave to come back close to its starting spot. The decay of this quantity with time is shown to change dramatically near the Anderson localization transition. The singular value decomposition of the propagation operator reveals the dominance of very intense recurrent scattering paths near the mobility edge.

Pulmonary Embolism Rule-out Criteria vs D-dimer testing in low-risk patients for pulmonary embolism: a retrospective study.

STUDY OBJECTIVES: The Pulmonary Embolism Rule-out Criteria (PERC) score has shown excellent negative predictive value; however, its use in the European population with high prevalence of PE is controversial. In Europe, PERC is not part of routine practice. For low-risk patients, guidelines recommend D-dimer testing, followed if positive by imaging study. We aimed to study the rate of diagnosis of PE after D-dimer testing in PERC-negative patients that could have been discharged if PERC was applied. METHOD: This was a multicenter retrospective study in Paris, France. We included all patients with a suspicion of PE who had D-dimer testing in the emergency department, low pre-test probability, and a negative PERC score (that was retrospectively calculated). Patients with insufficient record to calculate PERC score were excluded. The primary end point was the rate of PE diagnosis before discharge in this population. Secondary end points included rate of invasive imaging studies and subsequent adverse events. RESULTS: We screened 4301 patients who had D-dimer testing, 1070 of whom were PERC negative and could be analyzed. The mean age was 35 years and 46% were men. D-dimer was positive (>500 ng/L) in 167 (16%) of them; CTPA or V/Q scan was performed in 153 (14%) cases. PE was confirmed in 5 cases (total rate 0.5%, 95% confidence interval 0.1%-1.1%). Fifteen patients (1%) experienced non-severe adverse events. CONCLUSION: D-dimer testing in PERC-negative patients led to a diagnosis of PE in 0.5% of them, with 15% of patients undergoing unnecessary irradiative imaging studies.

Comparison of greenhouse gas emissions from sheep measured using both respiration and portable accumulation chambers.

Methane (CH4) is a potent gas produced by ruminants, and new measurement techniques are required to generate large datasets suitable for genetic analysis. One such technique are portable accumulation chambers (PAC), a short-term sampling method. The objectives of the current study were to explore the relationship between CH4 and carbon dioxide (CO2) output measured using both PAC and respiration chambers (RC) in growing lambs, and separately investigate the relationship among CH4, CO2 and measured ad libitum DM intake (DMI). Methane, CO2 and DMI were measured on 30 Suffolk and 30 Texel ewe lambs (age 253 ± 12 days) using the RC and PAC sequentially. The experiment was conducted over a 14-day period, with DMI measured from days 1 to 14; measurements in RC were conducted from days 10 to 12, while measurements in PAC were taken twice, the day immediately prior to the lambs entering the RC (day 9; PAC Pre-RC) and on the day lambs exited the RC (day 13; PAC Post-RC). Greater CH4 and CO2 output was measured in the RC than in the PAC (P < 0.01); similarly mean CH4 yield was greater when measured in the RC (15.39 ± 0.452 g CH4/kg DMI) compared to PAC (8.01 ± 0.767 g CH4/kg DMI). A moderate correlation of 0.37 was found between CH4 output measured in PAC Pre-RC and the RC, the corresponding regression coefficient of CH4 output measured in the RC regressed on CH4 output measured in PAC Pre-RC was close to unity (0.74; SE 0.224). The variance of CH4 and CO2 output within the measurement technique did not differ from each other (P > 0.05). Moderate to strong correlations were found between CH4 and CO2 per kg of live weight and CH4 and CO2 yield. Results from this study highlight the suitability of PAC as a ranking tool to rank animals based on their gaseous output when compared to the RC. However, repeated measurements separated by several days may be beneficial if precise rankings are required. Given the close to unity regression coefficient of CH4 output measured in the RC regressed on CH4 output measured in PAC Pre-RC suggests that PAC could also be potentially used to estimate absolute CH4 output; however, further research is required to substantiate this claim. When DMI is unknown, CH4 and CO2 per kg of live weight are a suitable alternative to the measurement of CH4 and CO2 yield.

Long-term control of vancomycin-resistant Enterococcus faecium at the scale of a large multihospital institution: a seven-year experience.

Repeated outbreaks of vancomycin-resistant Enterococcus faecium (VRE) occurred between 2004 and 2010 in Assistance Publique--Hôpitaux de Paris (AP-HP), a 23,000-bed multi-hospital institution. From August 2004 to December 2005, the French guidelines for preventing cross-transmission of multiresistant bacteria were applied. Because the number of VRE cases continued to increase, an institutional control programme was implemented from January 2006 onwards: it foresees stopping transfer of VRE and contact patients, separating VRE and contact patients in distinct cohorts, intervention of a central infection control team to support local teams, and quick application of measures as soon as first VRE cases are identified. Between August 2004 and December 2010, 45 VRE outbreaks occurred in 21 of the 38 AP-HP hospitals, comprising 533 cases. Time series analysis showed that the mean number of cases increased by 0.8 cases per month (95% confidence interval (CI): 0.3 to 1.3, p=0.001) before, and decreased by 0.7 cases per month after implementation of the programme (95% CI: -0.9 to -0.5, p<0.001), resulting in a significant trend change of -1.5 cases per month (95% CI: -2.1 to -0.9, p<0.001). The number of cases per outbreak was significantly lower after implementation of the programme. A sustained and coordinated strategy can control emerging bacteria at the level of a large regional multihospital institution.

Drug resistance in leprosy: An update following 70years of chemotherapy.

Leprosy is one of the oldest infectious diseases, reported for more than 2000years. Leprosy elimination goal as a public health problem set by the World Health Organization, aiming for a global prevalence rate<1 patient in a population of 10,000, was achieved in 2000 mainly thanks to the worldwide use of leprosy drugs starting in the 1980s and their access at no cost for patients since 1995. However, around 200,000 new cases are still reported each year, particularly in India, Brazil, and Indonesia. As with other bacteria of medical interest, antimicrobial resistance is observed in Mycobacterium leprae strains in several parts of the world, despite multidrug therapy being the recommended standard leprosy treatment to avoid resistance selection since 1982. Therefore, identifying and monitoring resistance is necessary. We provide an overview of the historical facts that led to the current drug resistance situation, the antibiotics effective against M. leprae, their mechanisms of action and resistance, and resistance detection methods. We also discuss therapeutic management of the resistant cases, new genes with potential roles in drug resistance and bacterial adaptation, new drugs under investigation, and the risk for resistance selection with the chemoprophylaxis measures.

Machine learning to improve the interpretation of intercalating dye-based quantitative PCR results.

This study aimed to evaluate the contribution of Machine Learning (ML) approach in the interpretation of intercalating dye-based quantitative PCR (IDqPCR) signals applied to the diagnosis of mucormycosis. The ML-based classification approach was applied to 734 results of IDqPCR categorized as positive (n = 74) or negative (n = 660) for mucormycosis after combining "visual reading" of the amplification and denaturation curves with clinical, radiological and microbiological criteria. Fourteen features were calculated to characterize the curves and injected in several pipelines including four ML-algorithms. An initial subset (n = 345) was used for the conception of classifiers. The classifier predictions were combined with majority voting to estimate performances of 48 meta-classifiers on an external dataset (n = 389). The visual reading returned 57 (7.7%), 568 (77.4%) and 109 (14.8%) positive, negative and doubtful results respectively. The Kappa coefficients of all the meta-classifiers were greater than 0.83 for the classification of IDqPCR results on the external dataset. Among these meta-classifiers, 6 exhibited Kappa coefficients at 1. The proposed ML-based approach allows a rigorous interpretation of IDqPCR curves, making the diagnosis of mucormycosis available for non-specialists in molecular diagnosis. A free online application was developed to classify IDqPCR from the raw data of the thermal cycler output ( http://gepamy-sat.asso.st/ ).

Dual nature of magnetic nanoparticle dispersions enables control over short-range attraction and long-range repulsion interactions.

Competition between attractive and repulsive interactions drives the formation of complex phases in colloidal suspensions. A major experimental challenge lies in decoupling independent roles of attractive and repulsive forces in governing the equilibrium morphology and long-range spatial distribution of assemblies. Here, we uncover the 'dual nature' of magnetic nanoparticle dispersions, particulate and continuous, enabling control of the short-range attraction and long-range repulsion (SALR) between suspended microparticles. We show that non-magnetic microparticles suspended in an aqueous magnetic nanoparticle dispersion simultaneously experience a short-range depletion attraction due to the particulate nature of the fluid in competition with an in situ tunable long-range magnetic dipolar repulsion attributed to the continuous nature of the fluid. The study presents an experimental platform for achieving in situ control over SALR between colloids leading to the formation of reconfigurable structures of unusual morphologies, which are not obtained using external fields or depletion interactions alone.

Exploiting the time-reversal operator for adaptive optics, selective focusing, and scattering pattern analysis.

We report on the experimental measurement of the backscattering matrix of a weakly scattering medium in optics, composed of a few dispersed gold nanobeads. The decomposition of the time-reversal operator is applied to this matrix and we demonstrate selective and efficient focusing on individual scatterers, even through an aberrating layer. Moreover, we show that this approach provides the decomposition of the scattering pattern of a single nanoparticle. These results open important perspectives for optical imaging, characterization, and selective excitation of nanoparticles.

Effect of a high-fat meal on the pharmacokinetics of dapagliflozin, a selective SGLT2 inhibitor, in healthy subjects.

Dapagliflozin is a potent and selective inhibitor of sodium-glucose co-transporter type 2 that is being developed for the treatment of type 2 diabetes mellitus. This open-label, randomized, two-period, two-treatment (single doses of 10-mg dapagliflozin fasted or fed), crossover study was conducted to evaluate the effect of a high-fat meal on the pharmacokinetics of dapagliflozin in 14 healthy subjects. Compared to the fasted state, a high-fat meal decreased mean dapagliflozin maximum plasma concentrations (C(max) ) by 31%, increased the time to C(max) (T(max) ) by 1 h, but did not affect overall dapagliflozin systemic exposure [area under the plasma concentration-time curve (AUC)]. As the cumulative (daily) amount of glucose excreted in the urine induced by dapagliflozin is dependent upon dapagliflozin AUC, the effect of food on dapagliflozin C(max) is unlikely to have a clinically meaningful effect on dapagliflozin's efficacy. On the basis of these findings, dapagliflozin can be administered without regard to meals.

[Histopathological study of Mycobacterium marinum infection]. / Aspects histopathologiques de l'infection à Mycobacterium marinum.

INTRODUCTION: Skin infection by Mycobacterium marinum induces the classic granuloma of aquariums and swimming pools. The histopathological signs have been described primarily in small series of typical cases, generally with no bacteriological evidence. In a national survey of proven infection with M. marinum detailed data was collected for 63 patients. The aim of this new study was to describe microscopic signs of the infection based upon biopsies taken from these patients. PATIENTS AND METHODS: Unstained slides from 32 biopsies of the skin (n=24) or synovial biopsies (n=8) were prepared; they originated from 27 patients. They were examined after standard staining and after Ziehl-Neelsen staining, without knowledge of the clinical data. RESULTS: All biopsies were taken from the upper limb of 18 men and nine women of mean age 48 years. Tubercular granulomas were observed in only 60% of cases. The largest and most numerous were seen in the synovial samples. Due to their palisade appearance, they were occasionally impossible to distinguish from rheumatoid nodules. In 20% of cases, neutrophil collections were seen without granulomas and in remaining 20% of cases, relatively non-specific infiltrate was observed. Epidermal changes consisted in psoriasiform or pseudocarcinomatous hyperplasia, particularly at the edges of ulcerated areas; invasion of the dermo-epidermal junction was seen in five cases. Follicular necrosis was observed in four cases with lymphoplasmacytic infiltrates remote from the granulomas being seen in 22 biopsies. Ziehl-Neelsen staining revealed no bacilli. DISCUSSION: The originality of this series consists of bacteriological proof of M. marinum infection and the absence of biopsy selection based on clinical criteria. It shows that the typical granulomas are in fact present in less than two third of cases, and that these may be confused with rheumatoid nodules. The chief characteristic of these lesions is the very low concentration of microorganisms present, in contrast with other forms of mycobacterium, making them difficult to see; routine confirmation cannot thus be expected from specific staining procedures. In one case out of five, the infiltrate suggested no infectious origin, although deep skin biopsies and synovial biopsies provided more information. For all forms of necrotic granuloma, whether or not accompanied by collections of neutrophils, a culture should be carried out in a specific medium, even in the absence of microscopic evidence of bacilli.

First evaluation of the automated-multiplex-PCR Unyvero ITI G2 cartridge for rapid diagnosis of osteo-articular infections.

OBJECTIVE: Conventional microbiological methods (CMM), including long-term culture, for the diagnosis of osteo-articular infections (OAI) fail in at least 5% of all cases. Only one IOA dedicated molecular method has been commercialized, and only the first version of this kit has been studied. The aim of this work was to evaluate the concordance between test results obtained with the second version of the Unyvero ITI G2 cartridge (Curetis) and CMM. The cartridge, combining one-step automated lysis/DNA extraction with multiplex PCR and amplicon detection by array hybridization, allows for the detection of 102 prevalent pathogens and their antibiotic resistance markers directly in clinical specimens (liquid [n=8] or solid [n=32]). MATERIAL AND METHODS: Frozen samples from 40 patients who underwent orthopedic surgery at Pitié-Salpêtrière hospital were tested retrospectively with the cartridge: 5 were culture-negative, 25 revealed monomicrobial and 10 polymicrobial OAI. The 2 main surgical sites were hip (22.5%) and knee (17.5%). RESULTS: Extraction, amplification and hybridization reactions were completed in 28 of the 40 cases, failed in all cartridge chambers in 6 cases, and in 1 or 2 chambers in an additional 6 cases. Overall sensitivity and specificity for microorganism identification were estimated at 67.6% and 98.2%, when complete and partial failures were excluded. CONCLUSIONS: These results show that the performances of the second version of the Unyvero ITI G2 cartridge should be further enhanced before considering avoiding conventional microbiological methods.

Sampling strategy for bacteriological diagnosis of intrathoracic tuberculosis.

BACKGROUND: Pulmonary tuberculosis (TB) is the most frequent site of TB and the one leading its spread worldwide. Multiple specimens are commonly collected for TB diagnosis including those requiring invasive procedures. This study aimed to review the sampling strategy for the microbiological diagnosis of pulmonary TB. METHODS: A retrospective analysis of collected samples from September 1st 2014 to May 1st 2016 in the Bacteriology laboratory of Pitié-Salpêtrière Hospital (Paris, France) was performed. All the samples collected in patients aged over 18 years for the bacteriological diagnosis of pulmonary TB were included. RESULTS: A total of 6267 samples were collected in 2187 patients. One hundred and twenty-six patients (6%) had a culture confirmed pulmonary TB. Among them, multiple sputum collections were sufficient for TB diagnosis in 63.5%, gastric lavages permitted to avoid bronchoscopy in only 7.1%, and bronchoscopy was necessary in 29.4%. The culture positivity of sputa (8.6%) was higher than that of bronchial aspirations (3.1%), bronchiolo-alveolar lavages (BAL) (2.3%) or gastric lavages (4.8%) (P<0.001). From its 70.0% theoretical PPV value, the 46.1% selection in bronchial aspirations allocated to molecular test increased PPV up to 88.9%. CONCLUSIONS: Based on our data, we suggest to collect sputum consistently. If smear negative a bronchoscopy should be performed and molecular diagnosis be performed on a subset of bronchial aspirations based on expertise of the bronchoscopist.

A processed pseudogene codes for a new antigen recognized by a CD8(+) T cell clone on melanoma.

The M88.7 T cell clone recognizes an antigen presented by HLA B*1302 on the melanoma cell line M88. A cDNA encoding this antigen (NA88-A) was isolated using a library transfection approach. Analysis of the genomic gene's sequence identified it is a processed pseudogene, derived from a retrotranscript of mRNA coding for homeoprotein HPX42B. The NA88-A gene exhibits several premature stop codons, deletions, and insertions relative to the HPX42B gene. In NA88-A RNA, a short open reading frame codes for the peptide MTQGQHFLQKV from which antigenic peptides are derived; a stop codon follows the peptide's COOH-terminal Val codon. Part of the HPX42B mRNA's 3' untranslated region codes for a peptide of similar sequence (MTQGQHFSQKV). If produced, this peptide can be recognized by M88.7 T cells. However, in HPX42B mRNA, the peptide's COOH-terminal Val codon is followed by a Trp codon. As a result, expression of HPX42B mRNA does not lead to antigen production. A model is proposed for events that participated in creation of a gene coding for a melanoma antigen from a pseudogene.

T cell response to Epstein-Barr virus transactivators in chronic rheumatoid arthritis.

Rheumatoid arthritis is a multistep disorder associated with autoimmune features of yet unknown etiology. Implication of viruses such as Epstein-Barr virus (EBV) in rheumatoid arthritis pathogenesis has been suspected on the basis of several indirect observations, but thus far, a direct link between EBV and rheumatoid arthritis has not been provided. Here we show that a large fraction of T cells infiltrating affected joints from a patient with chronic rheumatoid arthritis recognizes two EBV transactivators (BZLF1 and BMLF1) in a major histocompatibility complex-restricted fashion. Responses to these EBV antigens by synovial lymphocytes from several other chronic rheumatoid arthritis patients were readily detectable. Thus these results suggest a direct contribution of EBV to chronic rheumatoid arthritis pathogenesis. They also demonstrate for the first time the occurrence of T cell responses against EBV transactivating factors, which might be central in the control of virus reactivation.