Haemophilus influenzae one day in Denmark: prevalence, circulating clones, and dismal resistance to aminopenicillins.

Haemophilus influenzae is a common cause of mucosal infections that warrants accurate surveillance. We aimed to assess the prevalence of the species in clinical specimens, and characterise population structure and resistance to aminopenicillins by whole genome sequencing.We assessed the point prevalence by entering the database records of 1 day in Denmark and examined the genome sequences of nationwide, collected isolates from the same day. The prevalence of H. influenzae in clinical samples on the 10th of January 2018 was 1.78 per 100,000 person-days (all samples), and 2.47 per 1000 hospital bed-days (hospital samples). Of 2009 bacteria deemed clinically relevant and collected in a concerted action by the Danish departments of clinical microbiology, 62 (3.1%) were H. influenzae. All 62 isolates belonged to phylogenetic group I and were unencapsulated. Three strains from separate Danish regions had identical core genome sequences, but a small number of intergenic mutations testified to circulating clones, rather than individual cases of patient-to-patient transmission. The TEM-1 ß-lactamase gene was present in 24 strains, while 13 strains were genetically categorised as ampicillin-resistant due to substitutions in penicillin-binding protein 3; shared patterns of amino acid substitutions in unrelated strains indicated putative lateral transfer of chromosomal resistance. Circulating clones of H. influenzae are frequent, and host factors, rather than direct transmission of epidemic strains, may be the primary cause of infection. The bleak presence of ampicillin resistance revealed by sequencing of point prevalence strains underscores the necessity for close examination of testing methods.

Pain is frequent in children with cerebral palsy and negatively affects physical activity and participation.

AIM: The aim was to identify the prevalence of long-lasting pain among children with cerebral palsy (CP) and to investigate the association between pain and participation in physical leisure activities. METHODS: This is a cross-sectional study based on data from the National Danish Clinical Quality Database of children with CP. The study population consisted of 960 children aged 2-11 years across all Gross Motor Function Classification System (GMFCS) levels. Data were collected at children's regular clinical visits in 2016 or 2017. Information about pain and participation in physical leisure activities were obtained. The association was estimated as odds ratios (OR) and 95% confidence intervals (95% CI) by logistic regression adjusted for age and sex. RESULTS: We included data from 817 children (59% boys) median age 6 years, 52% classified as GMFCS level I. A total of 36% reported pain, and the most frequent pain locations were hips, feet and knees. Children reporting pain had lower odds for participation in physical leisure activities (OR 0.71, 95% CI 0.53-0.96). CONCLUSION: A large proportion of children with CP reported pain. There is an indication that long-lasting pain influences participation in physical leisure activities. Pain-relieving interventions are important to decrease pain-related suffering and facilitate participation.

Differences in life stage sensitivity of the beetle Tenebrio molitor towards a pyrethroid insecticide explained by stage-specific variations in uptake, elimination and activity of detoxifying enzymes.

It is widely accepted that sensitivity towards pesticides varies significantly between species. Much less is known about the potential differences in pesticide sensitivity and its biological mechanism throughout the lifecycle of a single species. In the present study we used three life-stages (larvae, pupae and adult) of the holometabolous insect Tenebrio molitor to investigate: i) Life-stage specific differences in sensitivity towards the pyrethroid insecticide α-cypermethrin after topical exposure, and ii) whether these differences can be explained by the degree of uptake and/or excretion. Finally, we investigated if an efficient excretion coincided with higher activities of the detoxifying enzymes cytochrome P450 (P450), esterases (EST) and glutathione-S-transferease (GST). We found that mobility of adults of T. molitor was more affected by α-cypermethrin treatment than larvae and pupae. Mortality was relatively low for all life stages and did not vary significantly with dose within the duration of the experiment, which indicated that death was (at least partly) due to starvation (indirect effect of paralysis) rather that direct effects of the insecticide. Insecticide treatment during the pupal stage further impaired normal development from pupa to adult. Toxicokinetic measurements showed that cuticle penetration of α-cypermethrin differed significantly between life-stages. Approximately 50% of the applied insecticide had penetrated the adult cuticle after 1 h, whereas a maximum of 30% and 16% had penetrated the waxier cuticle of larvae and pupae. Further, the pupal stage lacked the ability to excrete compounds, and hence internal insecticide concentrations in pupae increased or stagnated until emergence of the adult. Finally, quantification of detoxification enzymes showed a markedly higher activity of P450 in adults and larvae compared to pupae. These findings suggest that assessing toxicity and/or risk of pesticides collectively for a species may not be adequate without taking into account the potential sensitivity differences between life stages.

An increase in regulatory T cells in peripheral blood correlates with an adverse prognosis for malignant melanoma patients - A study of T cells and natural killer cells.

Malignant melanoma is a highly immunogenic tumour, and the immune profile significantly influences cancer development and response to immunotherapy. The peripheral immune profile may identify high risk patients. The current study showed reduced levels of CD4+ T cells and increased levels of CD8+ T cells in peripheral blood from malignant melanoma patients compared with controls. Percentages of peripheral CD56dimCD16+ NK cells were reduced and CD56brightCD16-KIR3+ NK cells were increased in malignant melanoma patients. Late stage malignant melanoma was correlated with low levels of CD4+ T cells and high levels of CD56brightCD16-KIR3+ NK cells. Finally, high levels of Tregs in peripheral blood were correlated with poor overall survival and disease-free survival. The results indicate that changes in specific immune cell subsets in peripheral blood samples from patients at the time of diagnosis may be potential biomarkers for prognosis and survival. Further studies will enable clarification of independent roles in tumour pathogenesis.

Are hip fracture patients with high or low body mass index at higher risk of missed care? A cohort study.

AIM: To examine whether patients' body mass index is associated with missed hip fracture care consistent with national guideline-recommended care. DESIGN: A nationwide, population-based cohort study using prospectively collected data from the Danish Multidisciplinary Hip Fracture Registry. METHODS: The study population consisted of 39,835 patients ≥65 years admitted with a hip fracture and discharged between 1st of January 2012 and 29th of November 2017. National guideline-recommended care consists of preoperative optimization, early surgery, mobilization within 24 h, basic mobility assessment, nutrition screening, post-discharge rehabilitation program, and osteoporotic and fall prophylaxis. We used binomial regression to estimate the relative risk for the fulfilment of the individual measures with 95% confidence interval. Multiple imputation method was applied to handle missing values of body mass index. RESULTS: The overall fulfilment of the individual measures ranged from 43% for pre-operative optimization to 95% for receiving a post-discharge rehabilitation program. The obese patients had a lower fulfilment of surgery within 36 h compared to patients with normal weight. No differences in fulfilment of the other measures were found. However, patients with missing data on body mass index had the highest risk of missed care. In conclusion, patients with missing BMI values had the highest risk of missed care. The obese patients had a slightly higher risk of long waiting times for surgery than normal-weighted patients. NO PATIENT OR PUBLIC CONTRIBUTION: This study was done based on population-based data from medical registries and data was analysed by the authors only.

Effect of Supplementing Dairy Goat Diets With Rapeseed Oil or Sunflower Oil on Performance, Milk Composition, Milk Fatty Acid Profile, and in vitro Fermentation Kinetics.

The aim of this study was to determine the effect of supplementing dairy goat diets with rapeseed oil and sunflower oil on performance, milk composition, milk fatty acid profile, and in vitro fermentation kinetics. Nine Danish Landrace goats with 42 ± 5 days in milk were allocated to three treatment groups for 42 days. Animals received a basal diet, formulated with 85:15 forage:concentrate ratio, and the basal diet was supplemented with either rapeseed oil or sunflower oil at 4% of dry matter. Goat milk was sampled on days 14, 21, and 42. Milk composition was similar between treatments. From day 14 to day 42, milk yield increased (1.03 vs. 1.34 kg/d), while milk fat (2.72 vs. 1.82 g/d) and total solids (11.2 vs. 9.14 %) were reduced. Compared to control and rapeseed oil, sunflower decreased (P < 0.05) C4:0 (1.56, and 1.67 vs. 1.36 g/100 g) and both oils decreased (P < 0.05) C18:3n3 (0.60 vs. 0.20 and 0.10 g/100g). Rapeseed oil increased (P < 0.05) C18:2 cis9, trans11 compared to control and sunflower oil (0.37 vs. 0.13 and 0.19 g/100 g). Untargeted milk foodomics revealed slightly elevated (P < 0.05) gluconic acid and decreased hippuric acid (P < 0.05) in the milk of oil-fed goats compared to control. In vitro dry matter degradation (63.2 ± 0.02 %) was not affected by dietary treatments, while individual volatile fatty acid proportions, total volatile fatty acids (35.7 ± 2.44 mmol/l), CO2 (18.6 ± 1.15 mol), and CH4 (11.6 ± 1.16 mol) were not affected by dietary treatments. Sunflower oil and rapeseed oil decreased (P < 0.05) total gas production at 24 and 48 h compared with control. Overall, the use of sunflower oil or rapeseed oil at 4% DM inclusion did not compromise animal performance and milk composition.

Influences of summer warming and nutrient availability on Salix glauca L. growth in Greenland along an ice to sea gradient.

The combined effects of climate change and nutrient availability on Arctic vegetation growth are poorly understood. Archaeological sites in the Arctic could represent unique nutrient hotspots for studying the long-term effect of nutrient enrichment. In this study, we analysed a time-series of ring widths of Salix glauca L. collected at nine archaeological sites and in their natural surroundings along a climate gradient in the Nuuk fjord region, Southwest Greenland, stretching from the edge of the Greenlandic Ice Sheet in the east to the open sea in the west. We assessed the temperature-growth relationship for the last four decades distinguishing between soils with past anthropogenic nutrient enrichment (PANE) and without (controls). Along the East-West gradient, the inner fjord sites showed a stronger temperature signal compared to the outermost ones. Individuals growing in PANE soils had wider ring widths than individuals growing in the control soils and a stronger climate-growth relation, especially in the inner fjord sites. Thereby, the individuals growing on the archaeological sites seem to have benefited more from the climate warming in recent decades. Our results suggest that higher nutrient availability due to past human activities plays a role in Arctic vegetation growth and should be considered when assessing both the future impact of plants on archaeological sites and the general greening in landscapes with contrasting nutrient availability.

Goat Milk Foodomics. Dietary Supplementation of Sunflower Oil and Rapeseed Oil Modify Milk Amino Acid and Organic Acid Profiles in Dairy Goats.

The dietary supplementation of vegetable oils is known to improve the dietary energy density as well as milk fatty acid profile; however, the impacts on the milk foodome is largely unknown. This study investigated the effect of two different sources of unsaturated fatty acids, rapeseed oil and sunflower oil, as a feeding supplement on the milk foodome from dairy goats. Nine Danish Landrace goats at 42 ± 5 days in milk were allocated to three treatment groups for 42 days with three animals per group. A control group received a basal diet made of forage and concentrate at an 85:15 ratio. On top of the basal diet, the second and third groups received rapeseed oil or sunflower oil supplements at 4% of dry matter, respectively. Goat milk was sampled on days 14, 21, and 42. The milk foodome was measured using gas chromatography-mass spectrometry and proton nuclear magnetic resonance spectroscopy. The milk levels of 2-hydroxyisovaleric acid, oxaloacetic acid, and taurine were higher in the milk from goats fed with sunflower oil compared to the control group. More glucose-1-phosphate was found in the milk from goats fed with rapeseed oil compared to the control group. Amino acids, valine and tyrosine, and 2-hydroxyisovaleric acid and oxaloacetic acid were higher in the sunflower group compared to the rapeseed group, while the milk from the rapeseed-fed goats had greater levels of ethanol and 2-oxoglutaric acid compared to the sunflower group. Thus, results show that foodomics is suitable for studying how milk chemistry changes as a function of feeding regime.

Chemical signature of Eurois occulta L. outbreaks in the xylem cell wall of Salix glauca L. in Greenland.

Insect defoliations are a major natural disturbance in high-latitude ecosystems and are expected to increase in frequency and severity due to current climatic change. Defoliations cause severe reductions in biomass and carbon investments that affect the functioning and productivity of tundra ecosystems. Here we combined dendro-anatomical analysis with chemical imaging to investigate the direct and lagged effects of insect outbreaks on carbon investment. We analysed the content of lignin vs. holocellulose, i.e. unspecified carbohydrates in xylem samples of Salix glauca L. collected at Iffiartarfik, Nuuk fjord, Greenland, featuring two outbreak events of the moth Eurois occulta L. Cross sections of the growth rings corresponding to both outbreaks ±3 years were analysed using confocal Raman imaging to identify possible chemical signatures related to insect defoliation on fibres, vessels, and ray parenchyma cells and to get insight into species-specific defence responses. Outbreak years with narrower rings and thinner fibre cell walls are accompanied by a change in the content of cell-wall polymers but not their underlying chemistry. Indeed, during the outbreaks the ratio between lignin and carbohydrates significantly increased in fibre but not vessel cell walls due to an increase in lignin content coupled with a reduced content of carbohydrates. Parenchyma cell walls and cell corners did not show any significant changes in the cell-wall biopolymer content. The selective adjustment of the cell-wall composition of fibres but not vessels under stressful conditions could be related to the plants priority to maintain an efficient hydraulic system rather than mechanical support. However, the higher lignin content of fibre cell walls formed during the outbreak events could increase mechanical stiffness to the thin walls by optimizing the available resources. Chemical analysis of xylem traits with Raman imaging is a promising approach to highlight hidden effects of defoliation otherwise overlooked with classical dendroecological methods.

Fungal attack on archaeological wooden artefacts in the Arctic-implications in a changing climate.

Climate change is expected to accelerate the microbial degradation of the many extraordinary well-preserved organic archaeological deposits found in the Arctic. This could potentially lead to a major loss of wooden artefacts that are still buried within the region. Here, we carry out the first large-scale investigation of wood degradation within archaeological deposits in the Arctic. This is done based on wooden samples from 11 archaeological sites that are located along a climatic gradient in Western Greenland. Our results show that Ascomycota fungi are causing extensive soft rot decay at all sites regardless of climate and local environment, but the group is diverse and many of the species were only found once. Cadophora species known to cause soft rot in polar environments were the most abundant Ascomycota found and their occurrence in native wood samples underlines that they are present locally. Basidiomycota fungi were also present at all sites. In the majority of samples, however, these aggressive and potentially very damaging wood degraders have caused limited decay so far, probably due to unfavorable growth conditions. The presence of these wood degrading fungi suggests that archaeological wooden artefacts may become further endangered if climate change leads to more favorable growth conditions.