Differential cytokine gene expression profiles in the three pathological forms of sheep paratuberculosis

<p>Abstract</p> <p>Background</p> <p>Johne's disease is a chronic inflammatory disease of the gut caused by infection with <it>Mycobacterium avium </it>subspecies <it>paratuberculosis </it>(MAP). Symptoms include wasting, diarrhoea, loss of condition and eventual death. Three forms of Johne's disease have been described in sheep – paucibacillary, multibacillary and asymptomatic. The paucibacillary form is characterized by an inflammatory, Th1-type immune response. The multibacillary form of the disease, which disseminates the infection, is characterized by macrophage infiltration mediated by a Th2-type immune response, and asymptomatic animals have no clinical symptoms or pathology but are infected with MAP. What determines these three forms of the disease is unknown. To further understand these differences, we used real-time RT-PCR to compare the expression of thirteen cytokine and cytokine-related genes in ileal tissue from sheep with the three forms of the disease.</p> <p>Results</p> <p>Three pathological forms of sheep paratuberculosis were defined on the basis of histopathology, cytochemistry (Zeihl-Neelsen) and IS900 PCR. Paucibacillary lesions have largely T cell and eosinophil infiltration and are ZN negative; multibacillary lesions have macrophage infiltration and large numbers of acid-fast bacteria. The pauci- and multibacillary forms are linked to the differential expression of IFNγ and IL-10 respectively. In addition the increased levels of the proinflammatory cytokines (IL-1β and TNFα), IL-8, IL-18 and TRAF-1 in both diseased forms is indicative of persistent inflammatory lesions. No changes were seen in IL-1α in any sheep ileum tissues. Asymptomatic animals are IS900+ with normal histology but have significantly decreased levels of IL-18 and increased levels TNFα.</p> <p>Conclusion</p> <p>We have quantified the expression levels of thirteen cytokine and cytokine related genes in three forms of ovine paratuberculosis using real-time PCR analyses and confirm that sheep pauci- and multibacillary disease are linked to type 1 and type 2 T cell responses respectively. The expression patterns of other cytokines shows that both disease forms have an inflammatory aetiology but that the central role for IL-1α in bovine paratuberculosis is not seen in the sheep infection. Asymptomatic animals are infected and show no pathology but can be distinguished, in terms of cytokine expression pattern, from uninfected controls.</p

Evaluation of a dual-scales method to measure weight-bearing through the legs, and effects of weight-bearing inequalities on hip bone mineral density and leg lean tissue mass.

Copyright © 2013 Foundation for Rehabilitation InformationOBJECTIVE: To investigate: the accuracy of measuring relative left/right weight-bearing using two identically calibrated weighing scales; the short-term weight-bearing tendencies in a general population of 9 participants and long-term in 42 females; the effect weight-bearing inequalities on hip bone mineral density and leg lean tissue mass. Method: Participants were measured standing astride two scales. Short-term volunteers were measured 10 times on one visit, with repositioning between measurements and the long-term group were measured on three visits at 6 month intervals. Baseline bilateral hip and total body Dual X-ray Absorptiometry scans were performed on the long-term group. RESULTS: The short-term Coefficient of Variation is 5.41% and long-term 7.01%. No significant correlations were found between hip bone density differences and weight-bearing inequalities, although a weak correlation of r = 0.31 (p = 0.047) was found for differences in leg lean tissue mass. CONCLUSION: Left/right weight-bearing measured using two scales is a consistent method for evaluating weight distribution through the legs. The short- and long-term weight-bearing tendencies showed a similar degree of variation. Weight-bearing inequalities were not associated with any significant left/right differences in bone mineral density at the hip, but were weakly associated with left-right differences in leg muscle mass

Duration of antibiotic treatment for common infections in English primary care: cross sectional analysis and comparison with guidelines

Objectives: To evaluate antibiotic therapy durations for common infections in English primary care and to compare this with guidelines. Design: Cross-sectional study. Setting: General practices contributing to The Health Improvement Network database, 2013-2015. Participants: 931,015 consultations that resulted in an antibiotic prescription for one of the following indications: acute sinusitis, acute sore throat, acute cough and bronchitis, pneumonia, acute exacerbation of chronic obstructive pulmonary disease (COPD), acute otitis media, acute cystitis, prostatitis, pyelonephritis, cellulitis, impetigo, scarlet fever and gastroenteritis. Main outcome measures: The main outcomes were the percentage of antibiotic prescriptions with a duration exceeding the guideline recommendation and the total number of days beyond the recommended duration for each indication. Results: The most common reasons for the prescriptions were patients consulting with acute bronchitis and cough (386,972), acute sore throat (239,231), acute otitis media (83,054), and acute sinusitis (76,683). Antibiotic treatments for upper respiratory indications and acute bronchitis accounted for more than two thirds of the total prescriptions considered, and ≥80% of these treatment courses exceeded guideline recommendations. Notable exceptions were acute sinusitis, where only 9.6% (95% CI 9.4 to 9.9%) of prescriptions exceeded 7 days and acute sore throat where only 2.1% (95% CI 2.0 to 2.1) exceed 10 days (recent guidance recommends 5 days). More than half of antibiotic prescriptions were longer than guidelines recommend for acute cystitis among females (54.6%, 95% CI 54.1 to 55.0%). The percentage of antibiotic prescriptions exceeding the recommended duration was lower for most non-respiratory infections. For the 931,015 included consultations resulting in antibiotic prescriptions, approximately 1.3 million days were beyond the durations recommended by the guidelines. Conclusion: For most common infections treated in primary care, a substantial proportion of antibiotic prescriptions have durations exceeding those recommended in guidelines. Substantial reductions in antibiotic exposure can be accomplished by aligning antibiotic prescription durations with guidelines

Spatial patterns and source attribution of urban methane in the Los Angeles Basin

Urban areas are increasingly recognized as a globally important source of methane to the atmosphere; however, the location of methane sources and relative contributions of source sectors are not well known. Recent atmospheric measurements in Los Angeles, California, USA, show that more than a third of the city's methane emissions are unaccounted for in inventories and suggest that fugitive fossil emissions are the unknown source. We made on-road measurements to quantify fine-scale structure of methane and a suite of complementary trace gases across the Los Angeles Basin in June 2013. Enhanced methane levels were observed across the basin but were unevenly distributed in space. We identified 213 methane hot spots from unknown emission sources. We made direct measurements of ethane to methane (C_2H_6/CH_4) ratios of known methane emission sources in the region, including cattle, geologic seeps, landfills, and compressed natural gas fueling stations, and used these ratios to determine the contribution of biogenic and fossil methane sources to unknown hot spots and to local urban background air. We found that 75% of hot spots were of fossil origin, 20% were biogenic, and 5% of indeterminate source. In regionally integrated air, we observed a wider range of C_2H_6/CH_4 values than observed previously. Fossil fuel sources accounted for 58–65% of methane emissions, with the range depending on the assumed C_2H_6/CH_4 ratio of source end-members and model structure. These surveys demonstrated the prevalence of fugitive methane emissions across the Los Angeles urban landscape and suggested that uninventoried methane sources were widely distributed and primarily of fossil origin

Inflammasome activation causes dual recruitment of NLRC4 and NLRP3 to the same macromolecular complex.

Pathogen recognition by nucleotide-binding oligomerization domain-like receptor (NLR) results in the formation of a macromolecular protein complex (inflammasome) that drives protective inflammatory responses in the host. It is thought that the number of inflammasome complexes forming in a cell is determined by the number of NLRs being activated, with each NLR initiating its own inflammasome assembly independent of one another; however, we show here that the important foodborne pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) simultaneously activates at least two NLRs, whereas only a single inflammasome complex is formed in a macrophage. Both nucleotide-binding domain and leucine-rich repeat caspase recruitment domain 4 and nucleotide-binding domain and leucine-rich repeat pyrin domain 3 are simultaneously present in the same inflammasome, where both NLRs are required to drive IL-1β processing within the Salmonella-infected cell and to regulate the bacterial burden in mice. Superresolution imaging of Salmonella-infected macrophages revealed a macromolecular complex with an outer ring of apoptosis-associated speck-like protein containing a caspase activation and recruitment domain and an inner ring of NLRs, with active caspase effectors containing the pro-IL-1β substrate localized internal to the ring structure. Our data reveal the spatial localization of different components of the inflammasome and how different members of the NLR family cooperate to drive robust IL-1β processing during Salmonella infection.S.M.M was supported by a Cambridge International Scholarship. T.P.M was supported by a Wellcome Trust Research Career Development Fellowship (WT085090MA). This study was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grants (BB/H003916/1 and BB/K006436/1) and a BBSRC Research Development Fellowship (BB/H021930/1) awarded to C.E.B.http://www.pnas.org/content/early/2014/05/05/1402911111.abstrac

Long-term oral antibiotic use in people with acne vulgaris in UK primary care: a drug utilization study

BACKGROUND: The inappropriate use of antibiotics is understood to contribute to antimicrobial resistance. Oral antibiotics are regularly used to treat moderate-to-severe acne vulgaris. In practice, we do not know the typical length of oral antibiotic treatment courses for acne in routine primary care and what proportion of people receive more than one course of treatment following a new acne diagnosis. OBJECTIVES: To describe how oral antibiotics are prescribed for acne over time in UK primary care. METHODS: We conducted a descriptive longitudinal drug utilization study using routinely collected primary care data from the Clinical Practice Research Datalink GOLD (2004-2019). We included individuals (8-50 years) with a new acne diagnosis recorded between 1 January 2004 and 31 July 2019. RESULTS: We identified 217 410 people with a new acne diagnosis. The median age was 17 years [interquartile range (IQR) 15-25] and median follow-up was 4.3 years (IQR 1.9-7.6). Among people with a new acne diagnosis, 96 703 (44.5%) received 248 560 prescriptions for long-term oral antibiotics during a median follow-up of 5.3 years (IQR 2.8-8.5). The median number of continuous courses of antibiotic therapy (≥ 28 days) per person was four (IQR 2-6). The majority (n = 59 010, 61.0%) of first oral antibiotic prescriptions in those with a recorded acne diagnosis were between the ages of 12 and 18. Most (n = 71 544, 74.0%) first courses for oral antibiotics were for between 28 and 90 days. The median duration of the first course of treatment was 56 days (IQR 50-93 days) and 18 127 (18.7%) of prescriptions of ≥ 28 days were for < 6 weeks. Among people who received a first course of oral antibiotic for ≥ 28 days, 56 261 (58.2%) received a second course after a treatment gap of ≥ 28 days. The median time between first and second courses was 135 days (IQR 67-302). The cumulative duration of exposure to oral antibiotics during follow-up was 255 days (8.5 months). CONCLUSIONS: Further work is needed to understand the consequences of using antibiotics for shorter periods than recommended. Suboptimal treatment duration may result in reduced clinical effectiveness or repeated exposures, potentially contributing to antimicrobial resistance

Interactions Between Zooplankton and Karenia brevis in the Gulf of Mexico.

Blooms of the toxic dinoflagellate K. brevis are common in the Gulf of Mexico, yet no in situ studies of the interactions between zooplankton and K. brevis in the Gulf of Mexico have been conducted. Zooplankton numerical abundance, biomass and taxonomic composition of nonbloom and K. brevis bloom stations within the ECOHAB study area were compared. At nonbloom stations, the most important determinant species were Parvolcalanus crassirostris, Oithona colcarva and Paracalanus quasimodo at the 5-m isobath and P. quasimodo, O. colcarva and Oikopleura dioka at the 25-m isobath. There was considerable overlap between the 5 and 25-m isobaths, with 9 species contributing to the top 90% of numerical abundance at both isobaths. Within K. brevis blooms Acartia tonsa, Centropages velificatus, Temora turbinata, Evadne tergestina, O. colcarva, O. dioika, and P. crassirostris were consistently dominant. Variations between non-bloom and bloom assemblages were evident, including variations in numerical abundance and biomass and the reduction in numerical abundance of 3 key species. Calculated grazing pressure proved insufficient to terminate K. brevis blooms, despite occasional grazing hot spots

The European Centre for Disease Prevention and Control (ECDC) pilot point prevalence survey of healthcare-associated infections and antimicrobial use

A standardised methodology for a combined point prevalence survey (PPS) on healthcare-associated infections (HAIs) and antimicrobial use in European acute care hospitals developed by the European Centre for Disease Prevention and Control was piloted across Europe. Variables were collected at national, hospital and patient level in 66 hospitals from 23 countries. A patient-based and a unit-based protocol were available. Feasibility was assessed via national and hospital questionnaires. Of 19,888 surveyed patients, 7.1% had an HAI and 34.6% were receiving at least one antimicrobial agent. Prevalence results were highest in intensive care units, with 28.1% patients with HAI, and 61.4% patients with antimicrobial use. Pneumonia and other lower respiratory tract infections (2.0% of patients; 95% confidence interval (CI): 1.8-2.2%) represented the most common type (25.7%) of HAI. Surgical prophylaxis was the indication for 17.3% of used antimicrobials and exceeded one day in 60.7% of cases. Risk factors in the patient-based protocol were provided for 98% or more of the included patients and all were independently associated with both presence of HAI and receiving an antimicrobial agent. The patient-based protocol required more work than the unit-based protocol, but allowed collecting detailed data and analysis of risk factors for HAI and antimicrobial use.peer-reviewe

Burden of SARS-CoV-2 infection in healthcare workers during second wave in England and impact of vaccines : prospective multicentre cohort study (SIREN) and mathematical model

Funding: The study is funded by the Department of Health and Social Care (DHSC) and UK Health Security Agency (UKHSA; formally Public Health England), with contributions from the governments of Northern Ireland, Scotland, and Wales. Funding was also provided by the National Institute for Health Research (NIHR) as an Urgent Public Health Priority Study and through the Health Protection Research Unit in Healthcare Associated Infections and Antimicrobial Resistance (NIHR200915), a partnership between UKHSA and the University of Oxford.OBJECTIVE: To describe the incidence of, risk factors for, and impact of vaccines on primary SARS-CoV-2 infection during the second wave of the covid-19 pandemic in susceptible hospital healthcare workers in England. DESIGN: Multicentre prospective cohort study. SETTING: National Health Service secondary care health organisations (trusts) in England between 1 September 2020 and 30 April 2021. PARTICIPANTS: Clinical, support, and administrative staff enrolled in the SARS-CoV-2 Immunity and Reinfection Evaluation (SIREN) study with no evidence of previous infection. Vaccination status was obtained from national covid-19 vaccination registries and self-reported. MAIN OUTCOME MEASURE: SARS-CoV-2 infection confirmed by polymerase chain reaction. Mixed effects logistic regression was conducted to determine demographic and occupational risk factors for infection, and an individual based mathematical model was used to predict how large the burden could have been if vaccines had not been available from 8 December 2020 . RESULTS: During England's second wave, 12.9% (2353/18 284) of susceptible SIREN participants became infected with SARS-CoV-2. Infections peaked in late December 2020 and decreased from January 2021, concurrent with the cohort's rapid vaccination coverage and a national lockdown. In multivariable analysis, factors increasing the likelihood of infection in the second wave were being under 25 years old (20.3% (132/651); adjusted odds ratio 1.35, 95% confidence interval 1.07 to 1.69), living in a large household (15.8% (282/1781); 1.54, 1.23 to 1.94, for participants from households of five or more people), having frequent exposure to patients with covid-19 (19.2% (723/3762); 1.79, 1.56 to 2.06, for participants with exposure every shift), working in an emergency department or inpatient ward setting (20.8% (386/1855); 1.76, 1.45 to 2.14), and being a healthcare assistant (18.1% (267/1479); 1.43, 1.16 to 1.77). Time to first vaccination emerged as being strongly associated with infection (P<0.001), with each additional day multiplying a participant's adjusted odds ratio by 1.02. Mathematical model simulations indicated that an additional 9.9% of all patient facing hospital healthcare workers would have been infected were it not for the rapid vaccination coverage. CONCLUSIONS: The rapid covid-19 vaccine rollout from December 2020 averted infection in a large proportion of hospital healthcare workers in England: without vaccines, second wave infections could have been 69% higher. With booster vaccinations being needed for adequate protection from the omicron variant, and perhaps the need for further boosters for future variants, ensuring equitable delivery to healthcare workers is essential. The findings also highlight occupational risk factors that persisted in healthcare workers despite vaccine rollout; a greater understanding of the transmission dynamics responsible for these is needed to help to optimise the infection prevention and control policies that protect healthcare workers from infection and therefore to support staffing levels and maintain healthcare provision. TRIAL REGISTRATION: ISRCTN registry ISRCTN11041050.Publisher PDFPeer reviewe