Faecal microbiota signatures of IBD and their relation to diagnosis, disease phenotype, inflammation, treatment escalation and anti-TNF response in a European Multicentre Study (IBD-Character).

METHOD: We examined faecal samples, using the GA-map™ Dysbiosis Test, to associate gut microbiota composition with Crohn's disease (CD) and ulcerative colitis (UC) and to identify markers for future biomarker identification. We conducted a prospective case-control study (EU-ref. no. 305676) in an inception cohort of 324 individuals (64 CD, 84 UC, 116 symptomatic non-IBD controls and 44 healthy controls) across five European centres and examined 54 predetermined bacterial markers. We categorized patients according to the Montreal Classification and calculated the dysbiosis index (DI). Non-parametric tests were used to compare groups and the Bonferroni correction to adjust for multiple comparisons. RESULTS: The fluorescent signals (FSSs) for Firmicutes and Eubacterium hallii were lower in inflammatory bowel disease (IBD) vs. symptomatic controls (p<.05). FSS for Firmicutes, Lachnospiraceae, Eubacterium hallii and Ruminococcus albus/bromii were lower, whereas the signal for Bacteroides Fragilis was higher in UC vs. symptomatic controls (p<.05). FSS was higher for Bifidobacterium spp., Eubacterium hallii, Actinobacteria and Firmicutes among patients with ulcerative proctitis, compared to extensive colitis (p<.05). In CD, we observed no association with disease location. The DI correlated with faecal-calprotectin in both CD and in UC (p<.001). In terms of treatment escalation and anti-TNF response, differences were observed for some bacterial markers, but none of these associations were statistically significant. CONCLUSION: Our data reveal that the GA-map™ Dysbiosis Test holds the potential to characterize the faecal microbiota composition and to assess the degree of dysbiosis in new-onset IBD. On the other hand, our results cannot demonstrate any proven diagnostic or predictive value of this method to support clinical decision making.

High frequency of occupied attB regions in Norwegian Staphylococcus aureus isolates supports a two-step MRSA screening algorithm.

Rapid nucleic acid amplification tests for methicillin-resistant Staphylococcus aureus (MRSA) diagnostics commonly target the mec resistance gene, genes specific for S. aureus, and the integration site for the SCCmec resistance cassette, orfX. Due to poor specificity when these target genes are used individually, additional culture is required to verify positive results. The combination of these targets is useful, but the optimal algorithm may depend on the presence of the genetic markers in S. aureus isolates, as well as the prevalence of MRSA in a population. The aim of the present study was to identify a rapid, low-cost, and functional screening algorithm in order to reduce the response time for MRSA diagnostics. An in-house orfX-SCCmec polymerase chain reaction (PCR) assay was established and evaluated. The results were compared with an existing mec/nuc PCR assay and traditional culture. Methicillin-sensitive S. aureus (MSSA) that tested false-positive in the orfX-SCCmec PCR assay were further investigated with full genome sequencing using the Ion PGM™ System to verify results and causality. Based on these data, a two-step screening algorithm with initial mec/nuc PCR followed by orfX-SCCmec PCR on positive samples was suggested and tested on 1443 patient samples. 22.5 % of MSSA isolates tested false-positive with the orfX-SCCmec PCR. Full genome sequencing of these isolates identified genetic variation in the attB region of S. aureus, including empty cassette variants and non-mec SCC. The suggested two-step MRSA screening algorithm allowed us to report MRSA results for 95.6 % of all samples and 99 % of MRSA-negative samples after one day.

Measuring laboratory-based influenza surveillance capacity: development of the 'International Influenza Laboratory Capacity Review' Tool.

OBJECTIVES: The 2005 International Health Regulations (IHR 2005) emphasized the importance of laboratory capacity to detect emerging diseases including novel influenza viruses. To support IHR 2005 requirements and the need to enhance influenza laboratory surveillance capacity, the Association of Public Health Laboratories (APHL) and the Centers for Disease Control and Prevention (CDC) Influenza Division developed the International Influenza Laboratory Capacity Review (Tool). STUDY DESIGN: Data from 37 assessments were reviewed and analyzed to verify that the quantitative analysis results accurately depicted a laboratory's capacity and capabilities. METHODS: Subject matter experts in influenza and laboratory practice used an iterative approach to develop the Tool incorporating feedback and lessons learnt through piloting and implementation. To systematically analyze assessment data, a quantitative framework for analysis was added to the Tool. RESULTS: The review indicated that changes in scores consistently reflected enhanced or decreased capacity. The review process also validated the utility of adding a quantitative analysis component to the assessments and the benefit of establishing a baseline from which to compare future assessments in a standardized way. CONCLUSIONS: Use of the Tool has provided APHL, CDC and each assessed laboratory with a standardized analysis of the laboratory's capacity. The information generated is used to improve laboratory systems for laboratory testing and enhance influenza surveillance globally. We describe the development of the Tool and lessons learnt.

Different glycosylation pattern of human IgG1 and IgG3 antibodies isolated from transiently as well as permanently transfected cell lines.

The effector functions of IgG depend on the presence of carbohydrates attached to asparagine 297 in the Fc-portion. In this report, glycosylation profiles of recombinant wild-type and mutant IgG1 and IgG3 antibodies produced from three cell lines were analysed using LC-ESI-Orbitrap. Clear differences were detected between IgG1 and IgG3 glycoforms, where IgG1 generally contained fucosylated glycoforms, whilst IgG3 mainly were non-fucosylated. When using NS-0 and J558L cells for permanent transfection, IgG1 wt glycoforms differed between the two cell lines, whilst IgG3 wt glycoforms did not. Transiently transfected HEK 293E cells were used to produce IgG1 and IgG3 wt and mutants, affecting complement activation. Cell supernatants were harvested at early and late time points and analysed separately. IgGs harvested late showed simpler and less developed glycosylation structure compared to those harvested early. The IgG harvested early was slightly more effective in complement activation than those harvested late, whilst the antibody-dependent cell-mediated cytotoxicity was unaltered. Generally, the glycosylation pattern of the mutants tested, including a hinge truncate mutant of IgG3, did not differ significantly from the wild-type IgGs. The striking difference in glycosylation pattern of IgG1 compared to IgG3 therefore appears not to be due to the long hinge region of IgG3 (62 amino acids) relative to the IgG1 hinge region (15 amino acids). Furthermore, mutation variants at or near the C1q binding site showed similar glycosylation structure and difference in their complement activation activity observed earlier is thus most likely due to differences in protein structure only.

Proteomic profiling of salmon skin mucus for the comparison of sampling methods.

The epidermal mucus protects fish against harmful environmental factors and the loss of physiological metabolites and water. It is an efficient barrier between the fish and the biosphere. The integrity of the skin mucus is thus of vital importance for the welfare and survival of the fish. Since excreted proteins and small molecules in the mucus can mirror the health status of the fish, it is a valuable matrix for monitoring stress, pathogen exposure, and nutritional effects. Several methods for sampling epidermal mucus from different fish species have previously been described, but information about their efficiency or the comparability of mucus analyses is lacking. In the present study, skin mucus from farmed Atlantic salmon was therefore sampled by three methods, including absorption or wiping with tissue paper, and scraping with a blunt blade, and the mucus proteome was analyzed by ultra-high pressure liquid chromatography high-resolution mass spectrometry. The measured protein contents, numbers, compositions and the observed data quality were compared between sampling methods. Furthermore, functional annotation and classification of the identified proteins was performed. The results showed that the three skin mucus sample types differed qualitatively as well as quantitatively. The absorbed mucus was the least tainted by proteins resulting from damage inflicted to the fish epidermis by the sampling procedure. Wiped mucus showed a better protein yield than absorbed and delivered a larger proteome of identifiable proteins, with less contamination from epithelial proteins than observed for scraped mucus. We recommend that future research of mucus should use the absorption method in cases, where it is important that the mucus is devoid of proteins from the underlying epithelium, and the wiping method, when protein yield is crucial or when the proteome of the outer epithelium is of interest.

Potential nitrous oxide production by marine shellfish in response to warming and nutrient enrichment.

Bivalves facilitate microbial nitrogen cycling, which can produce nitrous oxide (N2O), a potent greenhouse gas. Potential N2O production by three marine bivalves (Mytilus edulis, Mercenaria mercenaria and Crassostrea virginica) was measured in the laboratory including responses to nitrogen (N) loading and/or warming over short-terms (up to 14 or 28 days). N additions (targeting 100 µM-N ammonium nitrate) or warming (22 °C) individually and in combination were applied with experimental controls (20 µM-N, 19 °C). N2O production rates were higher with N additions for all species, but warming lacked significant direct effects. Ammonium and nitrate concentrations varied but were consistent with nitrification as a potential N2O source for all bivalves. Highest N2O emissions (7.5 nmol N2O g-1 h-1) were from M. edulis under hypoxic conditions coincident with a drop in pH. Macro-epifauna on M. edulis did not significantly alter N2O production. Thus, under short-term hypoxic conditions, micro-organisms in M. edulis guts may be a particularly significant source of N2O.

Progress toward sustainable influenza vaccination in the Lao Peoples' Democratic Republic, 2012-2018.

Despite global recommendations for influenza vaccination of high-risk, target populations, few low and middle-income countries have national influenza vaccination programs. Between 2012 and 2017, Lao PDR planned and conducted a series of activities to develop its national influenza vaccine program as a part of its overall national immunization program. In this paper, we review the underlying strategic planning for this process, and outline the sequence of activities, research studies, partnerships, and policy decisions that were required to build Laos' influenza vaccine program. The successful development and sustainability of the program in Laos offers lessons for other low and middle-income countries interested in initiating or expanding influenza immunization.

Epidemiology and spa-type diversity of meticillin-resistant Staphylococcus aureus in community and healthcare settings in Norway.

BACKGROUND: There has been a marked increase in the incidence of meticillin-resistant Staphylococcus aureus (MRSA) during the past decade in Norway; a country with one of the lowest prevalence rates and an active 'search-and-destroy' policy applied to hospital settings. AIM: To characterize the trends of notification rates of community-associated (CA) and healthcare-associated (HA) MRSA in Norway, and explore the diversity and circulation of MRSA spa types within and outside healthcare settings. METHODS: A registry-based study on notified MRSA infections and colonizations was conducted in Norway between 2006 and 2015. The diversity and abundance of CA- and HA-MRSA spa types were compared using novel ecological diversity measures (Hill numbers). FINDINGS: During the study period, the monthly notification rate increased 6.9-fold and 1.8-fold among CA- and HA-MRSA, respectively; the increase was steeper among colonizations than infections. In both settings, the distribution of spa types was uneven, with a few dominant spa types and many singletons. The spa-type diversity of CA-MRSA was higher than HA-MRSA in terms of different types (685 vs 481), and increased during the study period. However, the diversity associated with the dominant spa types was similar and remained stable. A high overlap of spa types was estimated between the settings; spa-t002, t019 and t008 were the most common. CONCLUSION: The present findings suggest a strong connection between CA- and HA-MRSA epidemiology in Norway. If the fast-growing trend of CA-MRSA continues in the years to come, it may challenge current guidelines and infection control of MRSA in healthcare environments.

[Measurement of the status of trace elements in cattle using liver biopsy samples]. / Het bepalen van de status van de spoorelementenvoorziening bij rundvee met behulp van leverbiopten.

Serum, plasma, or urine samples are usually used for the measurement of the trace elements copper; zinc, iron, selenium, because these samples are easy to obtain; however; these samples are not always appropriate. For example, it is not possible to measure molybdenum, the major antagonist of copper; in blood or urine. Therefore measurement of trace elements in liver tissue is considered the gold standard. For the assessment of selenium the method of choice remains determination of glutathion peroxidase in erythrocytes and for the assessment of magnesium determination of magnesium in urine. We determined the accuracy and repeatability of measuring trace elements in liver biopsies and whole liver homogenates. The levels of trace elements measured were similar in both preparations (92% agreement). Liver biopsy in live animals is a relatively simple procedure but not common in The Netherlands. Reference levels of trace elements, classified as too low, low, adequate, high, and too high, were established on the basis of our research and information in the literature. In a second study we investigated the practical aspects of obtaining liver tissue samples and their use. Samples were collected from cattle on a commercial dairy farm. Liver biopsy provided additional information to that obtained from serum and urine samples. We prepared a biopsy protocol and a test package, which we tested on 14 farms where an imbalance of trace minerals was suspected. Biopsy samples taken from 4 to 6 animals revealed extreme levels of trace elements.

Discussion of "Evidence-based Health Informatics: How Do We Know What We Know?".

This article is part of a For-Discussion-Section of Methods of Information in Medicine about the paper "Evidence-based Health Informatics: How Do We Know What We Know?" written by Elske Ammenwerth [1]. It is introduced by an editorial. This article contains the combined commentaries invited to independently comment on the Ammenwerth paper. In subsequent issues the discussion can continue through letters to the editor. With these comments on the paper "Evidence-based Health Informatics: How do we know what we know?", written by Elske Ammenwerth [1], the journal seeks to stimulate a broad discussion on the challenges of evaluating information processing and information technology in health care. An international group of experts has been invited by the editor of Methods to comment on this paper. Each of the invited commentaries forms one section of this paper.

Oxygen delivery and consumption in surfactant-depleted newborn piglets.

OBJECTIVE: To study the relationship between oxygen (O2) delivery (DO2) and O2 consumption (VO2) in surfactant-depleted newborn piglets. DESIGN: Prospective animal study. SETTING: Hospital surgical research laboratory. SUBJECTS: Twenty-six anesthetized and ventilated newborn piglets. INTERVENTIONS: Twenty of the animals were subjected to repeated saline lung lavages, and then assigned to either the saline group or the L-NAME group. The other six animals without lavage were studied as the control group. Piglets in the L-NAME group and the control group received 3 mg/kg of N(omega)-nitro-L-arginine methyl ester (L-NAME, an inhibitor of NO synthase) i.v.; and those in the saline group received the same volume of saline i.v. MEASUREMENTS AND RESULTS: Cardiac output (CO) was measured and arterial and mixed venous blood gases were analyzed. DO2, O2 extraction ratio (O2ER) and VO2 were calculated. Plasma hypoxanthine was analyzed. In the lung lavaged groups, cardiac index, DO2 and VO2 decreased significantly after L-NAME i.v. but not after saline i.v. Further, the decrease in VO2 in the L-NAME group correlated with the decrease in DO2 (r = 0.83, p < 0.001). In the control group, cardiac index and DO2, but not VO2, decreased significantly after L-NAME i.v. Simultaneously, O2ER increased significantly. Plasma hypoxanthine was not modified by lung lavage but increased after L-NAME i.v. in both the L-NAME and control groups. CONCLUSION: These data suggest that O2 supply dependency is present in surfactant-depleted newborn piglets.

A modified model for projecting age-structured populations in random environments.

A discrete-time age-structured population model with vital rates linked to a stochastic environmental process was developed as a generalization of an existing model by making the explicit link between variability in the vital rates and variability in the environment more flexible. This modified model uses biologically relevant probability distributions for the vital rates, and allows for temporal autocorrelation and an arbitrary covariance structure between vital rates. Through simulations, the properties of the projected population in the short-term were investigated and compared to analytical approximations. The distribution of the total population size did not quickly approach lognormality under all conditions. Furthermore, the sensitivity of the vital rates to the environmental process had a strong effect on the variance and distribution of the projected population size. These results suggest that short-term projections need to be carried out through simulation methods, as the analytical approximations technically apply only to the long-run asymptotic behavior. Techniques for parameter estimation were considered; recommendations depend on the form of the data available. The approach described allows the empirical calculation of the probability distribution for predicted population size, a quantity relevant to the use of formal decision analysis in natural resource management.

Bovine fetal neosporosis: a comparison of epizootic and sporadic abortion cases and different age classes with regard to lesion severity and immunohistochemical identification of organisms in brain, heart, and liver.

Eighty bovine fetuses with confirmed neosporosis were used to score lesion severity and presence of parasites in brain, heart, and liver. A comparison was made between epizootic and sporadic abortion cases. The possible influence of fetal age was also investigated. Histologic lesions of multifocal encephalitis, myocarditis, and periportal hepatitis with or without focal hepatocellular necrosis were almost always observed. Neospora caninum tachyzoites were identified immunohistochemically in 85% of the brains, 14% of the hearts, and 26% of the livers. Tissue cysts were observed in 21% of the brains. Significant differences between epizootic and sporadic abortion cases were found only in the liver. Hepatic lesions were more prominent and N. caninum tachyzoites were observed more frequently and in higher numbers in epizootic cases. Examination by immunohistochemistry of the liver in addition to the brain can be highly contributive diagnostically, particularly in epizootic cases. There were no significant age-related differences except for a higher presence of tachyzoites in the hearts of younger fetuses (3-4 months gestational age).

Characteristics of Neospora caninum-associated abortion storms in diary herds in The Netherlands (1995 to 1997).

Abortion storms in 50 dairy herds in The Netherlands were reported in which there was a strong association with Neospora caninum-infection. The duration of the abortion storms ranged from 6 to 65 d (mean 41.5 d). The cumulative proportion of aborting cows ranged from 0.11 to 0.57 (mean 0.26) of the animals at risk. An apparent seasonal influence was noted as most abortion storms occurred during the summer and early autumn. The prevalence of antibodies to N. caninum in 50 herds which had had an abortion storm was compared with that of 100 control herds which had no history of an abortion storm. Seroprevalence was estimated by testing a 20% cross sectional herd sample using a tachyzoite lysate-based ELISA method. Seroprevalence in case herds (range 17 to 87%, mean 51.5%) was significantly higher than that in control herds (range 0 to 53%, mean 13.9%). For most herds the seroprevalence levels were equal across all age groups, which suggests that the infection had been perpetuated by vertical transmission. In these herds, the abortion storms appeared to be induced by factors causing recrudescence of a N. caninum-infection in chronically infected animals rather than being the result of a recent introduction. In 6 case herds the seroprevalence in the dairy cows was significantly higher than in the young stock, which may have been attributable to superimposed postnatal infection.

Abortion risk in progeny of cows after a Neospora caninum epidemic.

A study was done of the descendants of cows from 4 dairy herds in which there had been N. caninum abortion outbreaks. Precolostral antibodies to N. caninum were demonstrated in 34 of 50 (68%) F1 calves and in 14 of 17 (82%) F2 calves from cows that aborted during the outbreaks. In 214 F1 progeny, N. caninum seroprevalence was nearly 50%, and there was a significant association between serostatus of the offspring and serostatus of dams. These observations indicated that congenital infection was an important mode of transmission after abortion outbreaks in these herds. A total of 52 abortions was recorded in 293 pregnancies of F1 progeny cows (1 to 3 pregnancies per animal). It was found that seropositive F1 cows had a three-fold increased abortion risk compared with seronegative F1 cows. In 2 of 10 abortions in seronegative cows evidence for N. caninum infection was found, suggesting that a low level of postnatal infection may also have occurred. It is concluded that N. caninum-infected calves should not be used as replacement stock, to decrease the future risk of abortion in dairy herds.

Increased risk of abortion following Neospora caninum abortion outbreaks: a retrospective and prospective cohort study in four dairy herds.

Explosive abortion outbreaks in 4 Dutch dairy herds during 1992 to 1994 are reported. In 50 of 51 fetuses submitted during the first 3 wk of the outbreaks characteristic histological lesions compatible for infection with Neospora caninum were seen. Diagnosis of infection was confirmed by immunohistochemistry in 40 fetuses (78%). No evidence for other abortifacients was found. The abortion risk of the herds was investigated in a prospective and retrospective cohort study. The prospective study showed that cows aborting during the outbreaks and N. caninum seropositive nonaborting cows had a two- to three-fold increased risk of abortion compared with N. caninum seronegative cows. Retrospective examination showed that seropositive nonaborting cows had an increased risk of abortion before the outbreaks, which may indicate that these animals were infected with N. caninum prior to the outbreaks. It is concluded that serostatus can be used for selective culling of cows to decrease future risk of abortion in dairy herds.

Graduate education in nursing leadership through distance technologies: the Canada-Norway Nursing Connection.

The Canada-Norway Nursing Connection was a collaborative project designed to provide an international educational experience for graduate students in nursing via distance technology. Computer-conferencing and video-teleconferencing were used to address nursing leadership content through case studies. The same technologies were employed to develop the project. The processes of planning and implementing the international linkage are described. Agreement about goals, content, context for online discussion, delivery methods, academic expectations, language support, and logistics was essential. The media proved to be effective for students to gain understandings about nursing leadership, health care, and the forces influencing the nursing profession globally. Insights from the project provided a basis for the development of a model for interactive, international graduate education that will be of value to educators dedicated to helping students gain a global understanding of nursing and health care issues.

Shifting the financial burden: the VA ambulatory care discharge policy.

The Department of Veterans Affairs is a primary source of health care services for many of the nation's uninsured and underinsured. Changes in congressionally mandated eligibility criteria and limited increases in appropriations have forced the Department to adopt a policy of discharging chronic but stable outpatients who have been treated for non-service-connected health conditions. Survey data from one VA medical center suggest that many, but not all, of those discharged: 1) have either Medicare or private insurance coverage; 2) have not sought or found alternative physician services in their local communities; 3) have discontinued taking previously prescribed medications; 4) report worsened health status since discharge; and, 5) have been hospitalized. In general, discharged patients from the lowest income group report the greatest financial access barriers. Preliminary analyses of the discharge policy suggest the potential for decreased access to needed medical services due to financial factors and cost-shifting from the VA to patients and other federal, state and local payers and providers.

spa typing alone is not sufficient to demonstrate endemic establishment of meticillin-resistant Staphylococcus aureus in a low-prevalence country.

BACKGROUND: The prevalence of meticillin-resistant Staphylococcus aureus (MRSA) in Norway is low but increasing. Over the last decade, numerous nursing homes have experienced MRSA outbreaks. One genetic lineage, spa type t304, has been identified at multiple nursing homes and has caused large outbreaks lasting for several years. AIM: To evaluate whether spa typing is sufficient for the detection of MRSA spread and endemic establishment in a low-prevalence area, using spa type t304 as the test organism. METHODS: All spa type t304 isolates detected in 1991-2010 in the most densely populated area of Norway were included. Time and place of bacterial sampling were recorded. The isolates were analysed using multi-locus sequence typing, staphylococcal cassette chromosome mec typing, detection of lukS/F-PV and pulsed-field gel electrophoresis (PFGE). FINDINGS: In total, 181 spa type t304 isolates were identified in three of 23 municipalities. Most (91%) of the isolates could be linked to 13 nursing homes, eight of which experienced outbreaks. PFGE analysis revealed three PFGE types, consisting of 19 PFGE patterns; 95% of the isolates were PFGE type 2. In total, PFGE types 2 and 3 accounted for 99% of all nursing home isolates, and included isolates from different nursing homes, different outbreaks and different time periods. Additional genetic analyses did not further differentiate between the spa type t304 isolates. CONCLUSION: MRSA spa type t304 appears to have established itself as an endemic genetic lineage in the study area. spa typing does not provide sufficient resolution when investigating the spread of an endemic-like genetic lineage in a low-prevalence area, and should be supplemented by additional typing techniques.

eHealth in Europe - Status and Challenges.

OBJECTIVES: To present European reflections on the concept of eHealth and emerging challenges related to further development of eHealth in Europe. METHODS: A survey with 10 questions was distributed to representatives of the national member associations of the European Federation of Medical Informatics (EFMI). RESULTS: The results document a shift from a constricting ICT-orientation to development of the entire health system where eHealth strategies, organizational change, and appropriate technological infrastructure are singled out as important aspects. CONCLUSION: There are urgent needs to ensure that eHealth strategies and policies for further design and deployment of eHealth applications support sociable services and innovations in health care.