Effects of feeding whole-cracked rapeseeds, nitrate, and 3-nitrooxypropanol on protein composition, minerals, and vitamin B in milk from Danish Holstein cows.

The present study was conducted to assess the individual or combined effects of feeding dietary fat (whole-cracked rapeseed), nitrate, and 3-nitrooxypropanol (3-NOP) on protein profile, mineral composition, B vitamins, and nitrate residues in milk from dairy cows. Forty-eight Danish Holstein cows used in an 8 × 8 incomplete Latin square design were fed 8 factorially arranged diets ((30 or 63 g crude fat/kg DM) × (0 or 10 g nitrate/kg DM) × (0 or 80 mg 3-NOP/kg DM)) over 6 periods of 21 d each. In each period, milk samples were collected from individual cows during the third week by pooling milk obtained from 4 consecutive milkings, and analyzed for protein profile including protein modifications, mineral composition, riboflavin, cobalamin, and presence of nitrate residues. Fat supplementation led to an increase in the phosphorylation degree of αS1-CN by 8.5% due to a decreased relative proportion of αS1-CN 8P and an increased relative proportion of αS1-CN 9P and further to a decrease in the relative proportion of αS2-CN by 2.4%. Additionally, fat supplementation decreased the relative proportions of glycosylated and unglycosylated forms of κ-CN, consequently leading to a 3.6% decrease in total κ-CN. In skim milk, K, Ca, P, and Mg concentrations were altered by individual use of fat, nitrate, and 3-NOP. Feeding nitrate resulted in a 5.4% increase in riboflavin concentration in milk while supplementing 3-NOP increased cobalamin concentration in milk by 21.1%. The nitrate concentration in milk was increased upon feeding nitrate however, this increased concentration was well below the maximum permissible limit of nitrate in milk (<50 mg/L). In conclusion, no major changes were observed in milk protein, and mineral compositions by feeding fat, nitrate, and 3-NOP to dairy cows while the increased riboflavin and cobalamin by nitrate and 3-NOP, respectively, could be of beneficial nutritional value for milk consumers.

The European Grief Conference, Copenhagen 2022: An effort to unite the field of bereavement care in Europe.

Bereavement care in Europe varies in quality and availability. Through greater collaboration across Europe, there could potentially be an opportunity to improve care. This article discusses the inaugural European Grief Conference held in Denmark in 2022: "Bereavement and Grief in Europe - Emerging Perspectives & Collaborations". The conference was structured around a 4-tiered public health model of bereavement care needs. It included practice, research, policy, and educational perspectives. A total of 250 people from 27 different countries participated. To determine if the conference had appealed to a broad European audience of grief professionals and to assess how the conference was received by participants, we examined registration/submission data, the results of a one-word real-time feedback exercise, and the answers to an online satisfaction survey. The results indicated wide interest in greater information sharing and collaboration across Europe among bereavement care, research, and education professionals.

Salivary fluoride levels after daily brushing with 5000 ppm fluoride toothpaste: A randomised, controlled clinical trial.

This study explored salivary fluoride levels following toothbrushing with 5000 and 1450 ppm fluoride toothpaste and determined the decline in salivary fluoride levels following the return from 5000 to 1450 ppm fluoride toothpaste. The study was a randomised, controlled double-blind parallel clinical trial (n = 24/group) measuring salivary fluoride five times during a 3-week trial phase involving 2×/day use of 5000 or 1450 ppm fluoride toothpaste, and five times during an ensuing 2-week wash-out phase where all participants used 1450 ppm toothpaste. Salivary fluoride was measured using a fluoride electrode and data were analysed using multilevel mixed-effects linear regression. Baseline salivary fluoride geometric means were 0.014 and 0.016 ppm for the 1450 and 5000 ppm groups, while the values at the end of the trial phase were 0.023 and 0.044 ppm, respectively. During the trial phase, except at baseline, differences between groups were statistically significant. The salivary fluoride levels for the 5000 ppm group remained statistically significantly higher than for the 1450 ppm group only at the first measurement in the wash-out phase (≈30 h after the last 5000 ppm brushing), indicating that higher salivary fluoride levels resulting from use of 5000 ppm are sustained only as long as the brushing habit continues.

Evidence of gene-gene interaction in hidradenitis suppurativa: a nationwide registry study of Danish twins.

BACKGROUND: Hidradenitis suppurativa (HS) is a recurrent inflammatory skin disease that, apart from rare causative loss-of-function mutations, has a widely unknown genetic aetiology. OBJECTIVES: To estimate the relative importance of genetic and environmental factors underlying susceptibility to HS. METHODS: Via the Danish Twin Registry and the Danish National Patient Registry we pulled together information on zygosity with that of HS status. Cases of HS were identified by the International Classification of Diseases (ICD)-8 (705·91) and ICD-10 (L73·2). Heritability was assessed by the classic biometric model and the possibility of gene-gene interaction via the multilocus modelling approach. RESULTS: Among 100 044 registered twins, we found 170 twins (from 163 pairs) diagnosed with HS. The seven concordant pairs were all monozygotic. Monozygotic twins had a case-wise concordance rate of 28% [95% confidence interval (CI) 7-49], corresponding to a familial risk of 73 (95% CI 13-133) times that of the background population. The biometrical modelling suggested a heritability of 0·80 (95% CI 0·67-0·93), and the multilocus index estimate was 230 (95% CI 60-400). This is highly indicative of gene-gene interactions, with the possibility of up to six interacting loci. CONCLUSIONS: This twin study was substantially larger and employed a more valid phenotype than previous studies. Genetics account for the majority of HS susceptibility, and HS is most likely caused by gene-gene interactions rather than monogenetic mutations or solely additive genetic factors. New approaches aimed at assessing potential interactions at a single-nucleotide polymorphism (SNP)-SNP level should be implemented in future HS genome-wide association studies.

Effect of initiating biologics compared to intensifying conventional DMARDs on clinical and MRI outcomes in established rheumatoid arthritis patients in clinical remission: Secondary analyses of the IMAGINE-RA trial.

OBJECTIVES: To compare the effect of treat-to-target-based escalations in conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) and biologics on clinical disease activity and magnetic resonance imaging (MRI) inflammation in a rheumatoid arthritis (RA) cohort in clinical remission. METHOD: One-hundred patients with established RA, Disease Activity Score based on 28-joint count-C-reactive protein (DAS28-CRP) < 3.2, and no swollen joints (hereafter referred to as 'in clinical remission') who received csDMARDs underwent clinical evaluation and MRI of the wrist and second to fifth metacarpophalangeal joints every 4 months. They followed a 2 year MRI treatment strategy targeting DAS28-CRP ≤ 3.2, no swollen joints, and absence of MRI osteitis, with predefined algorithmic treatment escalation: first: increase in csDMARDs; second: adding a biologic; third: switch biologic. MRI osteitis and Health Assessment Questionnaire (HAQ) (co-primary outcomes) and MRI combined inflammation and Simplified Disease Activity Index (SDAI) (key secondary outcomes) were assessed 4 months after treatment change and expressed as estimates of group differences. Statistical analyses were based on the intention-to-treat population analysed using repeated-measures mixed models. RESULTS: Escalation to first biologic compared to csDMARD escalation more effectively reduced MRI osteitis (difference between least squares means 1.8, 95% confidence interval 1.0-2.6), HAQ score (0.08, 0.03-0.1), MRI combined inflammation (2.5, 0.9-4.1), and SDAI scores (2.7, 1.9-3.5). CONCLUSIONS: Treat-to-target-based treatment escalations to biologics compared to escalation in csDMARDs more effectively improved MRI inflammation, physical function, and clinical disease activity in patients with established RA in clinical remission. Treatment escalation in RA patients in clinical remission reduces clinical and MRI-assessed disease activity. TRIAL REGISTRATION: Clinicaltrials.gov identifier: NCT01656278.

Characteristics of mammary secretions collected from infected and uninfected primigravid dairy heifer mammary glands.

Intramammary infections (IMI) in primigravid dairy heifers can affect mammary growth and development, which can reduce first-lactation milk yield. Detection of IMI in heifers most often involves the use of culture-based methods that are not often used in production dairy settings given their labor- and time-consuming nature. The objective of this study was to determine whether mammary secretion somatic cell count (SCC) and viscosity were associated with the infection status of primigravid heifer mammary glands. A total of 270 heifers from a single farm were used, selected based on the farmer's willingness to participate. The study was conducted from June to October 2020. Mammary secretion samples were aseptically collected from a randomly selected quarter of each heifer at 75 d prepartum (75PP), and another quarter of each heifer was sampled at 35 d prepartum (35PP). The remaining 2 quarters of each heifer were not examined. Mammary secretion samples underwent bacteriological examination to determine IMI status and quantitative SCC measurement and were also assessed for secretion viscosity based on visual observation. Prevalence of IMI was 26% (69/270) and 28% (71/255) at 75 and 35 d prepartum, respectively. Uninfected secretion samples had 133.2 [95% confidence interval (CI): 16.8 to >999.9] times greater odds to be thick compared with samples infected with a major pathogen, and 14.4 (95% CI: 8.5 to 24.1) times greater odds to be thick compared with samples infected with non-aureus staphylococci (NAS). The mean secretion SCC of uninfected quarters (6.04 ± 0.03 log10 cells/mL) was significantly lower than that of secretions collected from quarters infected with Staphylococcus chromogenes (6.34 ± 0.04 log10 cells/mL), other NAS species (6.28 ± 0.10 log10 cells/mL), or a major pathogen (6.73 ± 0.08 log10 cells/mL). These results indicate that mammary secretion viscosity and SCC measurement may be useful tools in identifying primigravid heifer quarters with IMI. The ability to evaluate viscosity at time of sampling may be a useful strategy that could be incorporated into interventions designed to diminish the negative effects of prepartum IMI on lactational performance.

Identification of rare genetic variants of the αS-caseins in milk from native Norwegian dairy breeds and comparison of protein composition with milk from high-yielding Norwegian Red cows.

Several factors influence the composition of milk. Among these, genetic variation within and between cattle breeds influences milk protein composition, protein heterogeneity, and their posttranslational modifications. Such variations may further influence technological properties, which are of importance for the utilization of milk into dairy products. Furthermore, these potential variations may also facilitate the production of differentiated products (e.g., related to specific breeds or specific genetic variants). The objective of this study was to investigate the genetic variation and relative protein composition of the major proteins in milk from 6 native Norwegian dairy breeds representing heterogeneity in geographical origin, using the modern Norwegian breed, Norwegian Red, as reference. In total, milk samples from 144 individual cows were collected and subjected to liquid chromatography-electrospray ionization/mass spectrometry-based proteomics for identification of genetic and posttranslational modification isoforms of the 4 caseins (αS1-CN, αS2-CN, ß-CN, κ-CN) and the 2 most abundant whey proteins (α-lactalbumin and ß-lactoglobulin). Relative quantification of these proteins and their major isoforms, including phosphorylations of αS1-CN and glycosylation of κ-CN, were determined based on UV absorbance. The presence and frequency of genetic variants of the breeds were found to be very diverse and it was possible to identify rare variants of the CN, which, to our knowledge, have not been identified in these breeds before. Thus, αS1-CN variant D was identified in low frequency in 3 of the 6 native Norwegian breeds. In general, αS1-CN was found to be quite diverse between the native breeds, and the even less frequent A and C variants were furthermore detected in 1 and 5 of the native breeds, respectively. The αS1-CN variant C was also identified in samples from the Norwegian Red cattle. The variant E of κ-CN was identified in 2 of the native Norwegian breeds. Another interesting finding was the identification of αS2-CN variant D, which was found in relatively high frequencies in the native breeds. Diversity in more common protein genetic variants were furthermore observed in the protein profiles of the native breeds compared with milk from the high-yielding Norwegian Reds, probably reflecting the more diverse genetic background between the native breeds.

Effects on feed intake, milk production, and methane emission in dairy cows fed silage or fresh grass with concentrate or fresh grass harvested at early or late maturity stage without concentrate.

The objective of the study was to quantify the effects on dry matter intake (DMI), nutrient digestibility, gas exchange, milk production, and milk quality in dairy cows fed fresh grass harvested at different maturity stages. Sixteen Danish Holstein cows in mid-lactation were divided into 4 blocks and used in 4 incomplete 4 × 2 Latin squares with 2 periods of 21 d. The cows received 1 of 4 treatments in each period, resulting in 8 cows per treatment, as follows: grass-clover silage supplemented with 6 kg/d concentrate pellets (SILc), fresh grass harvested at late maturity stage supplemented with 6 kg/d concentrate pellets (LATc), fresh grass harvested at late maturity stage (LAT), and fresh grass harvested at early maturity stage (ERL). The cows were housed in tiestalls and milked twice daily. The cows had ad libitum access to the forage, and concentrate pellets were divided into equal amounts and fed separately in the morning and afternoon. Fecal samples were collected to determine apparent total-tract digestibility, and samples of rumen fluid were collected for determination of short chain fatty acid composition. Halters were used for measuring eating and rumination time. Gas exchange was measured in open-circuit respiration chambers. Total DMI was higher in LATc and ERL (16.9 ± 0.45 and 15.5 ± 0.39 kg/d, respectively) compared with LAT (14.1 ± 0.42 kg/d). Relative to SILc, cows fed fresh grass experienced a convex pattern in DMI during the experiment. The changes in DMI were related to changes in leaf to stem ratio, fiber concentration, and organic matter digestibility determined in vitro in samples of the fresh grass harvested throughout the experiment. The apparent total-tract digestibility of organic matter was higher in SILc and LAT compared with LATc. Methane yield was lower for LATc compared with LAT (19.5 ± 0.61 vs. 22.6 ± 0.55 g of CH4/kg of DMI), and was not different between LAT and ERL. Compared with LAT, milk yield was higher for ERL (21.1 ± 1.14 vs. 23.4 ± 1.11 kg/d) and energy-corrected milk (ECM) yield was higher for LATc (21.5 ± 0.99 vs. 25.3 ± 1.03 kg/d). We detected no differences in milk or ECM yield between SILc and LATc. Milk protein yield was higher and milk fat concentration was lower in LATc compared with LAT. The fatty acid percentages of ∑C4-C14:1 and ∑C16 in milk were higher for SILc compared with LATc, signifying pronounced de novo synthesis. The n-6:n-3 ratio in milk fatty acids was lower for SILc and LAT compared with LATc, indicating improved nutritional quality for SILc and LAT. However, retinol concentration in milk was lower in SILc compared with all other treatments. The study implies that feeding silage instead of fresh grass has no effect on DMI, ECM yield, or CH4 yield, and that concentrate supplementation can increase milk production, affects milk quality, and reduces the effect on climate, whereas feeding less mature grass increases DMI and milk yield, but has no effect on CH4 yield.

Effect of protein concentrate mixtures and dietary addition of exogenous phytase on major milk minerals and proteins, including casein phosphorylation.

Variations in major milk minerals, proteins, and their posttranslational modifications are largely under genetic influence, whereas the effect of nongenetic factors is less studied. Through a controlled feeding experiment (incomplete balanced Latin square design), the effect of concentrate mixtures, based on fava beans, rapeseed meal, or soybean meal as main P and protein sources, on milk composition was examined under typical Danish management conditions. Concentrations of P, Ca, and Mg, together with proteomics for relative quantification of major milk proteins and their isoforms, were analyzed in milk samples from 24 cows sampled in 4 periods. Each cow was fed 1 of the 3 diets in each period with or without addition of exogenous phytase. Cows were blocked by lactation stage into early and mid-lactation (23.3 ± 6.7 and 176 ± 15 d in milk, respectively, at the beginning of the experiment, mean ± standard deviation). Significant effects of feed concentrate mixture were observed for milk protein concentration, milk urea nitrogen, citrate, and the percentage of mixed and preformed fatty acids as well as mineral composition, and their distributions within micellar or serum phases. Furthermore, relative contents of αS1-casein (CN) 9P form and unglycosylated κ-CN and thereby phosphorylation degree of αS1-CN (PD) and the glycosylation degree of κ-CN were found to be significantly affected by these diets. To our knowledge, we are the first to document that feed concentrate mixture can affect the relative concentrations of αS1-CN phosphorylation isoforms in milk, and the results suggested an effect on αS1-CN 9P and PD, but not on αS1-CN 8P. Furthermore, although only significant for αS1-CN 8P, we found a lower relative concentration of αS1-CN 8P and higher αS1-CN 9P (and thus higher PD) in milk from cows in mid compared with early lactation. Also, protein concentration and concentration of Mg in skim milk and serum as well as relative concentration of α-lactalbumin were found to be significantly affected by lactation stage. Addition of dietary exogenous phytase only had a minor effect on milk composition or functionality with significant effect detected for α-lactalbumin and micellar Mg concentration.

Genetic analysis on infrared-predicted milk minerals for Danish dairy cattle.

A group of milk components that has shown potential to be predicted with milk spectra is milk minerals. Milk minerals are important for human health and cow health. Having an inexpensive and fast way to measure milk mineral concentrations would open doors for research, herd management, and selective breeding. The first aim of this study was to predict milk minerals with infrared milk spectra. Additionally, milk minerals were predicted with infrared-predicted fat, protein, and lactose content. The second aim was to perform a genetic analysis on infrared-predicted milk minerals, to identify QTL, and estimate variance components. For training and validating a multibreed prediction model for individual milk minerals, 264 Danish Jersey cows and 254 Danish Holstein cows were used. Partial least square regression prediction models were built for Ca, Cu, Fe, K, Mg, Mn, Na, P, Se, and Zn based on 80% of the cows, selected randomly. Prediction models were externally validated with 8 herds based on the remaining 20% of the cows. The prediction models were applied on a population of approximately 1,400 Danish Holstein cows with 5,600 infrared spectral records and 1,700 Danish Jersey cows with 7,200 infrared spectral records. Cows from this population had 50k imputed genotypes. Prediction accuracy was good for P and Ca, with external R2 ≥ 0.80 and a relative prediction error of 5.4% for P and 6.3% for Ca. Prediction was moderately good for Na with an external R2 of 0.63, and a relative error of 18.8%. Prediction accuracies of milk minerals based on infrared-predicted fat, protein, and lactose content were considerably lower than those based on the infrared milk spectra. This shows that the milk infrared spectrum contains valuable information on milk minerals, which is currently not used. Heritability for infrared-predicted Ca, Na, and P varied from low (0.13) to moderate (0.36). Several QTL for infrared-predicted milk minerals were observed that have been associated with gold standard milk minerals previously. In conclusion, this study has shown infrared milk spectra were good at predicting Ca, Na, and P in milk. Infrared-predicted Ca, Na, and P had low to moderate heritability estimates.

Administration of internal teat sealant in primigravid dairy heifers at different times of gestation to prevent intramammary infections at calving.

Intramammary infections (IMI) are common in primigravid dairy heifers and can negatively affect future milk production. Bismuth subnitrate-based internal teat sealants (ITS) have been used to prevent prepartum IMI in dairy heifers by creating a physical barrier within the teat, preventing pathogens from entering the gland, though determination of when to administer ITS in heifers has yet to be investigated. The objectives of this study were to determine if administration of ITS in primigravid heifers reduced the odds of IMI at calving and if administration of ITS at different stages of gestation (75 vs. 35 d prepartum) affected the odds of IMI at calving. A total of 270 heifers were used at a single farm. One quarter of each heifer was randomly chosen to be aseptically sampled and administered ITS 75 d prepartum (ITS75), another quarter of each heifer was sampled and received ITS 35 d prepartum (ITS35), whereas the remaining 2 quarters of each heifer served as control quarters (CON) and were not sampled before calving. Within 12 h of calving, aseptic colostrum samples were collected from all quarters to determine quarter infection status. When an IMI was caused by mastitis pathogens other than non-aureus staphylococci (NAS), CON quarters were 3 times [95% confidence interval (CI): 1.4-6.3] and 2.5 times (95% CI: 1.2-4.9) more likely to be infected at calving than ITS75 and ITS35 quarters, respectively. For IMI with NAS, CON quarters were 5.8 (95% CI: 3.2-10.5) and 6.4 (95% CI: 3.4-12.0) times more likely to be infected than ITS75 and ITS35 quarters, respectively. Odds of IMI at calving was similar between ITS75 and ITS35 quarters for both NAS (odds ratio = 0.9) and other pathogens (odds ratio = 1.2). Results indicate that ITS administration at either 75 and 35 d prepartum reduced IMI prevalence at calving in primigravid dairy heifers. Farm specific factors may influence prevalence and timing of heifer IMI and earlier administration of ITS provides an extended period of protection for the developing gland.

Reduced mesiodistal tooth dimension in individuals with osteogenesis imperfecta: a cross-sectional study.

OBJECTIVE: Osteogenesis imperfecta (OI) is a rare, hereditary disease affecting collagen type-1 in connective tissue. Collagen type-1 is a substantial component of dentine, and it is speculated, whether affected dentine could cause altered mesiodistal tooth dimension possibly affecting restorative treatment regimen. Therefore, the aim of the present study was to measure mesiodistal tooth dimensions in individuals with OI and compare them with healthy controls. MATERIALS AND METHODS: Fifty-seven individuals aged 20-77 years with OI type 1-4 were included and 70 control patients aged 11-34 years were drawn from an orthodontic database. Mesiodistal tooth dimensions of all tooth types, except third molars, were measured in mm (two decimals) on digital 3 D-models of the tooth-bearing arches. RESULTS: Multilevel mixed-effects linear regression analysis showed that mesiodistal tooth dimension on average was 0.17 mm (95% CI = (-0.33; -0.01)) reduced for the OI group compared to controls. The analysis revealed variation between tooth types; incisors and first premolars were most affected and molars minimally affected. CONCLUSIONS: The mesiodistal tooth dimension in individuals diagnosed with OI is significantly smaller compared to healthy controls, which should be taken into consideration in the restorative treatment planning of individuals with OI, although the magnitude of the deviation is relatively small. The results on mesiodistal tooth dimensions of the present controls may be used as a standard for comparisons in future studies on tooth dimensions.

The transformation of a gold standard in-person substance use assessment to a web-based, REDCap integrated data capture tool.

The adoption of computer systems for gathering, managing, and analyzing health data is resulting in the replacement of pen-and-paper methods for collecting data and managing health records by computerized methods. One classic "pen-and-paper" assessment in health and substance use research is the Timeline Follow-Back (TLFB), the gold standard in self-reported substance use developed in 1996 by Sobell et al. to assess alcohol consumption patterns and later other substances such as marijuana or tobacco over discreet timeframes [1-7]. The TLFB has been modified by some research groups for use as a web-based assessment [8-10], but not without significant limitations. As such, this paper describes the team-oriented, interdisciplinary process by which a new online TLFB (O-TLFB) was conceptualized, the technical details of development towards a dynamic data capture tool fully integrated with REDCap via application programming interface (API), and the potential for this optimized O-TLFB to be leveraged broadly across the domains of substance use, health, and behavioral research.

Natural variations of citrate and calcium in milk and their effects on milk processing properties.

Natural variations among milk constituents, and their relations to each other as well as to processing parameters, represent possibilities for differentiation of milk to produce high-quality natural products. In this study, we focused on natural variations in milk citrate and its interplay with calcium distribution in milk, in relation to processing properties. Milk samples from individual cows from farms varying in feeding and management practices were collected from April to June 2017 to maximize natural variations in citrate and calcium. Chemical composition, rennet coagulation properties, and ethanol stability were analyzed for all milk samples. We focused particularly on calcium distribution and citrate content and the correlation of these to other milk parameters. No significant change in citrate content was observed during the sampling period, which suggests that mechanisms other than feeding affect citrate levels in milk. Several significant correlations were found, including a positive correlation between complexed serum calcium and citrate, and a negative correlation between urea and ionic calcium. These are both of interest in relation to further processing, as with regard to the stability of UHT milk and in cheese making. Although the correlation between complexed serum calcium and citrate may be explained by their affinity, the underlying driver for the negative relationship between natural milk urea and ionic calcium needs to be clarified by further studies. Furthermore, milk from the different farms varied not only with regard to organic versus conventional farming systems; feeding practices between farms also play an important role in milk composition and functionality. However, none of the differences in milk composition between farms were found to decrease milk functionality and thus would probably not cause any processing problems.

A comparative study on basophil activation test, histamine release assay, and passive sensitization histamine release assay in the diagnosis of peanut allergy.

BACKGROUND: Allergy can be diagnosed using basophil tests. Several methods measuring basophil activation are available. This study aimed at comparing basophil activation test (BAT), histamine release assay (HR), and passive sensitization histamine release assay (passive HR) in the diagnosis of peanut allergy. METHODS: BAT, HR, and passive HR were performed on 11 peanut-allergic and 14 nonallergic subjects. Blood was incubated with peanut extract or anti-IgE and tests were performed as follows: BAT-CD63 upregulation was assessed by flow cytometry; HR-released histamine was quantified by a glass fiber-based fluorometric method; passive HR-IgE-stripped donor basophils were incubated with participants' serum and histamine release was quantified as HR. RESULTS: CDsens, a measure of basophil allergen sensitivity, was significantly higher for BAT (80.1±17.4) compared to HR (23.4±10.31) and passive HR (11.1±2.0). BAT, HR, and passive HR had a clinical sensitivity of 100%, 100%, and 82% and specificity of 100%, 100%, and 100%, respectively, when excluding inconclusive results. BAT identified 11 of 11 allergic patients, HR 10, and passive HR 9. Likewise, BAT recognized 12 of 14 nonallergic subjects, HR 10, and passive HR 13. However, the tests' diagnostic performances were not statistically different. Interestingly, nonreleasers in HR but not in BAT had lower basophil count compared to releasers (249 vs 630 counts/min). CONCLUSION: BAT displayed a significantly higher CDsens compared to HR and passive HR. The basophil tests' diagnostic performances were not significantly different. Still, BAT could diagnose subjects with low basophil number in contrast to HR.

Prediction of unfavorable outcomes in cryptococcal meningitis: results of the multicenter Infectious Diseases International Research Initiative (ID-IRI) cryptococcal meningitis study.

Cryptococcal meningitis (CM) is mostly seen in immunocompromised patients, particularly human immunodeficiency virus (HIV)-positive patients, but CM may also occur in apparently immunocompetent individuals. Outcome analyses have been performed in such patients but, due to the high prevalence of HIV infection worldwide, CM patients today may be admitted to hospitals with unknown HIV status, particularly in underdeveloped countries. The objective of this multicenter study was to analyze all types of CM cases in an aggregate cohort to disclose unfavorable outcomes. We retrospectively reviewed the hospitalized CM patients from 2000 to 2015 in 26 medical centers from 11 countries. Demographics, clinical, microbiological, radiological, therapeutic data, and outcomes were included. Death, neurological sequelae, or relapse were unfavorable outcomes. Seventy (43.8%) out of 160 study cases were identified as unfavorable and 104 (65%) were HIV infected. On multivariate analysis, the higher Glasgow Coma Scale (GCS) scores (p = 0.021), cerebrospinal fluid (CSF) leukocyte counts > 20 (p = 0.038), and higher CSF glucose levels (p = 0.048) were associated with favorable outcomes. On the other hand, malignancy (p = 0.026) was associated with poor outcomes. Although all CM patients require prompt and rational fungal management, those with significant risks for poor outcomes need to be closely monitored.

Correction to: Prediction of unfavorable outcomes in cryptococcal meningitis: results of the multicenter infectious Diseases International Research Initiative (ID-IRI) cryptococcal meningitis study.

In the original version of this article, Mustafa Sunbul was not included in the list of authors for this article. The name has been added accordingly.

Evaluation of ELISA and haemagglutination inhibition as screening tests in serosurveillance for H5/H7 avian influenza in commercial chicken flocks.

Avian influenza virus (AIV) subtypes H5 and H7 can infect poultry causing low pathogenicity (LP) AI, but these LPAIVs may mutate to highly pathogenic AIV in chickens or turkeys causing high mortality, hence H5/H7 subtypes demand statutory intervention. Serological surveillance in the European Union provides evidence of H5/H7 AIV exposure in apparently healthy poultry. To identify the most sensitive screening method as the first step in an algorithm to provide evidence of H5/H7 AIV infection, the standard approach of H5/H7 antibody testing by haemagglutination inhibition (HI) was compared with an ELISA, which detects antibodies to all subtypes. Sera (n = 1055) from 74 commercial chicken flocks were tested by both methods. A Bayesian approach served to estimate diagnostic test sensitivities and specificities, without assuming any 'gold standard'. Sensitivity and specificity of the ELISA was 97% and 99.8%, and for H5/H7 HI 43% and 99.8%, respectively, although H5/H7 HI sensitivity varied considerably between infected flocks. ELISA therefore provides superior sensitivity for the screening of chicken flocks as part of an algorithm, which subsequently utilises H5/H7 HI to identify infection by these two subtypes. With the calculated sensitivity and specificity, testing nine sera per flock is sufficient to detect a flock seroprevalence of 30% with 95% probability.

Cerebral Metabolic Changes Related to Oxidative Metabolism in a Model of Bacterial Meningitis Induced by Lipopolysaccharide.

BACKGROUND: Cerebral mitochondrial dysfunction is prominent in the pathophysiology of severe bacterial meningitis. In the present study, we hypothesize that the metabolic changes seen after intracisternal lipopolysaccharide (LPS) injection in a piglet model of meningitis is compatible with mitochondrial dysfunction and resembles the metabolic patterns seen in patients with bacterial meningitis. METHODS: Eight pigs received LPS injection in cisterna magna, and four pigs received NaCl in cisterna magna as a control. Biochemical variables related to energy metabolism were monitored by intracerebral microdialysis technique and included interstitial glucose, lactate, pyruvate, glutamate, and glycerol. The intracranial pressure (ICP) and brain tissue oxygen tension (PbtO2) were also monitored along with physiological variables including mean arterial pressure, blood glucose, lactate, and partial pressure of O2 and CO2. Pigs were monitored for 60 min at baseline and 240 min after LPS/NaCl injection. RESULTS: After LPS injection, a significant increase in cerebral lactate/pyruvate ratio (LPR) compared to control group was registered (p = 0.01). This increase was due to a significant increased lactate with stable and normal values of pyruvate. No significant change in PbtO2 or ICP was registered. No changes in physiological variables were observed. CONCLUSIONS: The metabolic changes after intracisternal LPS injection is compatible with disturbance in the oxidative metabolism and partly due to mitochondrial dysfunction with increasing cerebral LPR due to increased lactate and normal pyruvate, PbtO2, and ICP. The metabolic pattern resembles the one observed in patients with bacterial meningitis. Metabolic monitoring in these patients is feasible to monitor for cerebral metabolic derangements otherwise missed by conventional intensive care monitoring.

The immunoglobulin superfamily member CD200R identifies cells involved in type 2 immune responses.

BACKGROUND: The pathology of allergic diseases involves type 2 immune cells, such as Th2, ILC2, and basophils exerting their effect by production of IL-4, IL-5, and IL-13. However, surface receptors that are specifically expressed on type 2 immune cells are less well documented. The aim of this investigation was to identify surface markers associated with type 2 inflammation. METHODS: Naïve human CD4+ T cells were short-term activated in the presence or absence of IL-4 and analyzed for expression of >300 cell-surface proteins. Ex vivo-isolated peripheral blood mononuclear cells (PBMCs) from peanut-allergic (PA) and nonallergic subjects were stimulated (14-16 h) with peanut extract to detect peanut-specific CD4+ CD154+ T cells. Biopsies were obtained for transcriptomic analysis from healthy controls and patients with extrinsic or intrinsic atopic dermatitis (AD) and psoriasis. RESULTS: Expression analysis of >300 surface proteins enabled identification of IL-4-upregulated surface proteins, such as CD90, CD108, CD109, and CD200R (CD200R1). Additional analysis of in vitro-differentiated Th0, Th1, and Th2 cultures identified CD200R as upregulated on Th2 cells. From ex vivo-isolated PBMCs, we found high expression of CD200R on Th2 and ILC2 cells and basophils. In PA subjects, the peanut-specific Th2 (CD154+ CRTh2+ ) cells expressed more CD200R than the non-allergen-specific Th2 (CD154- CRTh2+ ) cells. Moreover, costaining of CD161 and CD200R identified peanut-specific highly differentiated IL-4+ IL-5+ Th2 cells. Finally, transcriptomic analysis revealed upregulation of CD200R in lesional skin from subjects with an extrinsic AD phenotype compared to healthy skin. CONCLUSION: These results indicate that CD200R expression strongly correlates with Th2 pathology; though, the mechanism is as yet elusive.