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Cytokines, demyelination and neuroaxonal degeneration in the central nervous system are pivotal elements implicated in the pathogenesis of multiple sclerosis (MS) and its nonclinical model of experimental autoimmune encephalomyelitis (EAE). Phycocyanobilin (PCB), a chromophore of the biliprotein C-Phycocyanin (C-PC) from Spirulina platensis, has antioxidant, immunoregulatory and anti-inflammatory effects in this disease, and it could complement the effect of other Disease Modifying Treatments (DMT), such as Interferon-β (IFN-β). Here, our main goal was to evaluate the potential PCB benefits and its mechanisms of action to counteract the chronic EAE in mice. MOG35-55-induced EAE was implemented in C57BL/6 female mice. Clinical signs, pro-inflammatory cytokines levels by ELISA, qPCR in the brain and immunohistochemistry using precursor/mature oligodendrocytes cells antibodies in the spinal cord, were assessed. PCB enhanced the neurological condition, and waned the brain concentrations of IL-17A and IL-6, pro-inflammatory cytokines, in a dose-dependent manner. A down- or up-regulating activity of PCB at 1 mg/kg was identified in the brain on three (LINGO1, NOTCH1, and TNF-α), and five genes (MAL, CXCL12, MOG, OLIG1, and NKX2-2), respectively. Interestingly, a reduction of demyelination, active microglia/macrophages density, and axonal damage was detected along with an increase in oligodendrocyte precursor cells and mature oligodendrocytes, when assessed the spinal cords of EAE mice that took up PCB. The studies in vitro in rodent encephalitogenic T cells and in vivo in the EAE mouse model with the PCB/IFN-β combination, showed an enhanced positive effect of this combined therapy. Overall, these results demonstrate the anti-inflammatory activity and the protective properties of PCB on the myelin and support its use with IFN-β as an improved DMT combination for MS.

Abstract Background Parasitoid insects manipulate their hosts' physiology by injecting various factors into their host upon parasitization. Transcriptomic approaches provide a powerful approach to study insect host-parasitoid interactions at the molecular level. In order to investigate the effects of parasitization by an ichneumonid wasp (Diadegma semiclausum) on the host (Plutella xylostella), the larval transcriptome profile was analyzed using a short-read deep sequencing method (Illumina). Symbiotic polydnaviruses (PDVs) associated with ichneumonid parasitoids, known as ichnoviruses, play significant roles in host immune suppression and developmental regulation. In the current study, D. semiclausum ichnovirus (DsIV) genes expressed in P. xylostella were identified and their sequences compared with other reported PDVs. Five of these genes encode proteins of unknown identity, that have not previously been reported. Results De novo assembly of cDNA sequence data generated 172,660 contigs between 100 and 10000 bp in length; with 35% of > 200 bp in length. Parasitization had significant impacts on expression levels of 928 identified insect host transcripts. Gene ontology data illustrated that the majority of the differentially expressed genes are involved in binding, catalytic activity, and metabolic and cellular processes. In addition, the results show that transcription levels of antimicrobial peptides, such as gloverin, cecropin E and lysozyme, were up-regulated after parasitism. Expression of ichnovirus genes were detected in parasitized larvae with 19 unique sequences identified from five PDV gene families including vankyrin, viral innexin, repeat elements, a cysteine-rich motif, and polar residue rich protein. Vankyrin 1 and repeat element 1 genes showed the highest transcription levels among the DsIV genes. Conclusion This study provides detailed information on differential expression of P. xylostella larval genes following parasitization, DsIV genes expressed in the host and also improves our current understanding of this host-parasitoid interaction.

Abstract Background Escherichia coli is a model prokaryote, an important pathogen, and a key organism for industrial biotechnology. E. coli W (ATCC 9637), one of four strains designated as safe for laboratory purposes, has not been sequenced. E. coli W is a fast-growing strain and is the only safe strain that can utilize sucrose as a carbon source. Lifecycle analysis has demonstrated that sucrose from sugarcane is a preferred carbon source for industrial bioprocesses. Results We have sequenced and annotated the genome of E. coli W. The chromosome is 4,900,968 bp and encodes 4,764 ORFs. Two plasmids, pRK1 (102,536 bp) and pRK2 (5,360 bp), are also present. W has unique features relative to other sequenced laboratory strains (K-12, B and Crooks): it has a larger genome and belongs to phylogroup B1 rather than A. W also grows on a much broader range of carbon sources than does K-12. A genome-scale reconstruction was developed and validated in order to interrogate metabolic properties. Conclusions The genome of W is more similar to commensal and pathogenic B1 strains than phylogroup A strains, and therefore has greater utility for comparative analyses with these strains. W should therefore be the strain of choice, or 'type strain' for group B1 comparative analyses. The genome annotation and tools created here are expected to allow further utilization and development of E. coli W as an industrial organism for sucrose-based bioprocesses. Refinements in our E. coli metabolic reconstruction allow it to more accurately define E. coli metabolism relative to previous models.

Abstract Background The mortality in patients with persistent low CD4 count despite several years of HAART with sustained viral suppression is poorly documented. We aimed to identify predictors for inadequate CD4 cell recovery and estimate mortality in patients with low CD4 count but otherwise successful HAART. Method In a nationwide cohort of HIV patients we identified all individuals who started HAART before 1 January 2005 with CD4 cell count ≤ 200 cells/μL and experienced three years with sustained viral suppression. Patients were categorized according to CD4 cell count after the three years suppressed period (≤ 200 cells/μL; immunological non-responders (INRs), >200 cells/μL; immunological responders (IRs)). We used logistic regression and Kaplan-Meier analysis to estimated risk factors and mortality for INRs compared to IRs. Results We identified 55 INRs and 236 IRs. In adjusted analysis age > 40 years and > one year from first CD4 cell count ≤ 200 cells/μL to start of the virologically suppressed period were associated with increased risk of INR. INRs had substantially higher mortality compared to IRs. The excess mortality was mainly seen in the INR group with > one year of immunological suppression prior to viral suppression and injection drug users (IDUs). Conclusion Age and prolonged periods of immune deficiency prior to successful HAART are risk factors for incomplete CD4 cell recovery. INRs have substantially increased long-term mortality mainly associated with prolonged immunological suppression prior to viral suppression and IDU.

Abstract Background The quantitative analysis of metabolic fluxes, i.e., in vivo activities of intracellular enzymes and pathways, provides key information on biological systems in systems biology and metabolic engineering. It is based on a comprehensive approach combining (i) tracer cultivation on 13C substrates, (ii) 13C labelling analysis by mass spectrometry and (iii) mathematical modelling for experimental design, data processing, flux calculation and statistics. Whereas the cultivation and the analytical part is fairly advanced, a lack of appropriate modelling software solutions for all modelling aspects in flux studies is limiting the application of metabolic flux analysis. Results We have developed OpenFLUX as a user friendly, yet flexible software application for small and large scale 13C metabolic flux analysis. The application is based on the new Elementary Metabolite Unit (EMU) framework, significantly enhancing computation speed for flux calculation. From simple notation of metabolic reaction networks defined in a spreadsheet, the OpenFLUX parser automatically generates MATLAB-readable metabolite and isotopomer balances, thus strongly facilitating model creation. The model can be used to perform experimental design, parameter estimation and sensitivity analysis either using the built-in gradient-based search or Monte Carlo algorithms or in user-defined algorithms. Exemplified for a microbial flux study with 71 reactions, 8 free flux parameters and mass isotopomer distribution of 10 metabolites, OpenFLUX allowed to automatically compile the EMU-based model from an Excel file containing metabolic reactions and carbon transfer mechanisms, showing it's user-friendliness. It reliably reproduced the published data and optimum flux distributions for the network under study were found quickly ( Conclusion We have developed a fast, accurate application to perform steady-state 13C metabolic flux analysis. OpenFLUX will strongly facilitate and enhance the design, calculation and interpretation of metabolic flux studies. By providing the software open source, we hope it will evolve with the rapidly growing field of fluxomics.

Abstract Background Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a commensal of horses and an opportunistic pathogen in many animals and humans. Some strains produce copious amounts of hyaluronic acid, making S. zooepidemicus an important industrial microorganism for the production of this valuable biopolymer used in the pharmaceutical and cosmetic industry. Encapsulation by hyaluronic acid is considered an important virulence factor in other streptococci, though the importance in S. zooepidemicus remains poorly understood. Proteomics may provide a better understanding of virulence factors in S. zooepidemicus, facilitate the design of better diagnostics and treatments, and guide engineering of superior production strains. Results Using hyaluronidase to remove the capsule and by optimising cellular lysis, a reference map for S. zooepidemicus was completed. This protocol significantly increased protein recovery, allowing for visualisation of 682 spots and the identification of 86 proteins using mass spectrometry (LC-ESI-MS/MS and MALDI-TOF/TOF); of which 16 were membrane proteins. Conclusion The data presented constitute the first reference map for S. zooepidemicus and provide new information on the identity and characteristics of the more abundantly expressed proteins.

Abstract Background We examined whether impaired renal function causes thickening of the aortic valve leaflets in hyperlipidemic apoE-knockout (apoE-/-) mice, and whether the putative effect on the aortic valves could be prevented by inhibiting the angiotensin-converting enzyme (ACE) with enalapril. Methods Thickening of the aortic valve leaflets in apoE-/- mice was induced by producing mild or moderate chronic renal failure resulting from unilateral nephrectomy (1/2 NX, n = 18) or subtotal nephrectomy (5/6 NX, n = 22), respectively. Additionally, the 5/6 NX mice were randomized to no treatment (n = 8) or enalapril treatment (n = 13). The maximal thickness of each leaflet was measured from histological sections of the aortic roots. Results Leaflet thickness was significantly greater in the 5/6 NX mice than in the 1/2 NX mice (P = 0.030) or the unoperated mice (P = 0.003). The 5/6 NX mice treated with enalapril had significantly thinner leaflets than did the untreated 5/6 NX mice (P = 0.014). Conclusion Moderate uremia causes thickening of the aortic valves in apoE-/- mice, which can be attenuated by ACE inhibition. The nephrectomized apoE-/- mouse constitutes a new model for investigating the mechanisms of uremia-induced aortic valve disease, and also provides an opportunity to study its pharmacologic prevention.

Abstract Background Knowledge about the complete genome constellation of seasonal influenza A viruses from different countries is valuable for monitoring and understanding of the evolution and migration of strains. Few complete genome sequences of influenza A viruses from Europe are publicly available at the present time and there have been few longitudinal genome studies of human influenza A viruses. We have studied the evolution of circulating human H3N2, H1N1 and H1N2 influenza A viruses from 1999 to 2006, we analysed 234 Danish human influenza A viruses and characterised 24 complete genomes. Results H3N2 was the prevalent strain in Denmark during the study period, but H1N1 dominated the 2000–2001 season. H1N2 viruses were first observed in Denmark in 2002–2003. After years of little genetic change in the H1N1 viruses the 2005–2006 season presented H1N1 of greater variability than before. This indicates that H1N1 viruses are evolving and that H1N1 soon is likely to be the prevalent strain again. Generally, the influenza A haemagglutinin (HA) of H3N2 viruses formed seasonal phylogenetic clusters. Different lineages co-circulating within the same season were also observed. The evolution has been stochastic, influenced by small "jumps" in genetic distance rather than constant drift, especially with the introduction of the Fujian-like viruses in 2002–2003. Also evolutionary stasis-periods were observed which might indicate well fit viruses. The evolution of H3N2 viruses have also been influenced by gene reassortments between lineages from different seasons. None of the influenza genes were influenced by strong positive selection pressure. The antigenic site B in H3N2 HA was the preferred site for genetic change during the study period probably because the site A has been masked by glycosylations. Substitutions at CTL-epitopes in the genes coding for the neuraminidase (NA), polymerase acidic protein (PA), matrix protein 1 (M1), non-structural protein 1 (NS1) and especially the nucleoprotein (NP) were observed. The N-linked glycosylation pattern varied during the study period and the H3N2 isolates from 2004 to 2006 were highly glycosylated with ten predicted sequons in HA, the highest amount of glycosylations observed in this study period. Conclusion The present study is the first to our knowledge to characterise the evolution of complete genomes of influenza A H3N2, H1N1 and H1N2 isolates from Europe over a time period of seven years from 1999 to 2006. More precise knowledge about the circulating strains may have implications for predicting the following season strains and thereby better matching the vaccine composition.

Abstract Background The basophil activation test (BAT), in which translocation of markers to the surface of blood basophils is measured in response to allergen by flow cytometry, is a rapid assay that is gaining popularity. Two markers are currently being evaluated for the BAT; CD63 and the lineage-specific CD203c. In a recent report, detection of CD203c after lysis with Saponin was shown to be superior to detection of CD63 after lysis with formic acid. We wanted to compare a) lysis with formic acid and lysis with Saponin, b) the response through CD203c and CD63, and c) the definition 10% activated cells above background with the probability binning metric T(χ) > 4, on sets of data generated with blood basophils stimulated with varying concentrations of anti-FcεRI antibody. Methods Blood from volunteers was incubated with serial logarithmic dilutions of anti-FcεRI and subsequently with antibodies to CD203c PE and CD63 FITC. Sets of samples set up in parallel were lysed with either Saponin based Whole Blood Lysing reagent or with formic acid based Immunoprep/Q-prep. Samples were acquired on a FACS Calibur, but were compensated and analysed offline. Responders were defined as persons who had 10% or more activated basophils above background, or a T(χ) > 4, for two consecutive dilutions of anti-FcεRI antibody. Results More basophils (median 1164 vs. median 397) and better discrimination of upregulated CD203c and CD63 amongst responders were obtained after lysis with Saponin than after lysis with formic acid. We suggest that CD203c may be a more sensitive marker for the BAT than CD63, as 6/11 responders were found with CD203c, compared with 3/11 with CD63. Most responders (7/11) were identified with probability binning. Conclusion A combination of lysis with Saponin and the markers CD203c and CD63 computed by probability binning may be the most sensitive method of detecting activation of basophils after stimulation through FcεRI.

The mounting evidence that loneliness is a determinant of poor health and well-being underpins the need for effective interventions and community action. 'More Together' (MoTo) is a large-scale, complex, multi-component and multi-level intervention for community change that addresses loneliness among young people and older adults in Silkeborg Municipality, Denmark. The intervention is inspired by the Collective Impact framework, and it is practice driven and rooted in an extensive cross-sector partnership. This article outlines (i) the organization of the cross-sector partnership, (ii) the structure of the intervention programme, (iii) the key components and activities of the programme and, finally, (iv) the intervention setting and target population. MoTo aims to create new ways to develop, implement and evaluate loneliness interventions. Experiences gained from MoTo hold the potential to transform our understanding of loneliness interventions and may inform and guide future interventions., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: COVID-19 has highlighted the importance of patient activation in managing chronic conditions and promoting resilience during times of crisis. Patient activation refers to an individual's knowledge, skills, and confidence in managing their own health and healthcare. Previous research has shown that people with higher levels of patient activation are better prepared to navigate the challenges of chronic illness and are more likely to engage in healthy behaviors. However, the impact of patient activation on COVID-19-related concerns and mental well-being among people with chronic conditions during the pandemic remains unclear. This study aims to investigate the possible role of patient activation in shaping COVID-19-related concerns and to describe changes in mental well-being among Danish adults with one or more chronic conditions during the early months of the pandemic., Methods: Danish adults with chronic conditions (e.g. diabetes, coronary heart disease, obstructive pulmonary lung disease, cancer) who had participated in a municipal health education program prior to the COVID-19 outbreak were asked to participate in this prospective questionnaire study in May 2020 and November 2020. Sociodemographic (sex, age, living status, educational attainment, employment status) and disease-related information (diagnosis, one or more chronic conditions) along with the Patient Activation Measure were collected before the outbreak and were obtained from a clinical database used for monitoring and evaluation of municipal health education programs. In contrast, the two questionnaires collected six months apart consisted of single items related to concerns about COVID-19 and the WHO-5 well-being index., Results: A total of 710 people with chronic conditions (mean age 60.9 years; 55.8% female) participated at both time points. In bivariate analyses, patient activation was associated with COVID-19-related concern and well-being. At follow-up, participants experienced a significant decrease in well-being. The decrease was associated with poorer well-being measured six months earlier, a greater perception that it had become more challenging to take care of one's health due to the pandemic, and finally, feeling lonely. The association between patient activation and well-being ceased to be significant in the multivariate regression model., Conclusions: A considerable proportion of people with chronic conditions participating in this study have been mentally burdened during COVID-19. Although lower levels of patient activation were associated with greater COVID-19-related concerns, it did not have a significant impact on mental well-being over time., (© 2023. The Author(s).)

Background: With its health risks and extensive disruption to everyday life, the SARS-CoV-2 (COVID-19) pandemic has affected the lives of billions of people. People with chronic conditions are particularly susceptible to severe illness if infected by COVID-19, and they have repeatedly been urged to take stringent steps to 'shield' themselves from the virus. It is argued that the negative impact of isolation and other lockdown-related restrictions on emotional well-being and daily life may be most prominent among people at increased risk for severe illness from COVID-19. This qualitative thematic analysis aimed to explore how individuals with chronic conditions perceived the risk posed by COVID-19 and to understand how being at high risk affected their emotional well-being and everyday life., Methods: The study is a thematic analysis of qualitative data consisting of semi-structured interviews with adults affected by at least one chronic condition supplemented with free text comments from a PRO-based survey., Results: Based on 17 semi-structured interviews and 144 free text comments from a PRO-based survey three thematic patterns representing diverse COVID-19-related risk experiences were extracted: (1) Feeling vulnerable and at risk, (2) Uncertainty about being at risk, and (3) Distancing from the high-risk label., Conclusions: The risk of COVID-19 impacted the participants' everyday lives and emotional well-being in various ways. Some participants felt vulnerable and at risk causing them and their families to take on far-reaching precautions with significant consequences for their everyday life and emotional well-being. Some participants expressed uncertainty associated with whether they were at increased risk. Such uncertainty gave rise to dilemmas about how to navigate their everyday life. Other participants did not identify themselves as at higher risk and took no special precautions. Such a lack of perceived risk may undermine their motivation for taking preventive measures, which calls for public attention regarding current or future pandemics., (© 2023. The Author(s).)

Self-assembly and fibril formation play important roles in protein behaviour. Amyloid fibril formation is well-studied due to its role in neurodegenerative diseases and characterized by refolding of the protein into predominantly β-sheet form. However, much less is known about the assembly of proteins into other types of supramolecular structures. Using cryo-electron microscopy at a resolution of 1.97 Å, we show that a triple-mutant of the anti-microbial peptide plectasin, PPI42, assembles into helical non-amyloid fibrils. The in vitro anti-microbial activity was determined and shown to be enhanced compared to the wildtype. Plectasin contains a cysteine-stabilised α-helix-β-sheet structure, which remains intact upon fibril formation. Two protofilaments form a right-handed protein fibril. The fibril formation is reversible and follows sigmoidal kinetics with a pH- and concentration dependent equilibrium between soluble monomer and protein fibril. This high-resolution structure reveals that α/β proteins can natively assemble into fibrils., (© 2022. The Author(s).)

Enzymes can aid in optimal feed stock utilization when used as feed additives. A range of toxicological studies were performed to evaluate the safety profile of a novel phytase (phytase HM) from Citrobacter b raakii produced in Aspergillus oryzae . Phytase HM was found to be non-mutagenic and non-clastogenic in in vitro tests. Further, the phytase HM preparation did not exhibit irritative potential to the eye and skin when applied in in vitro models. A 13-week subchronic toxicity study with oral administration of phytase HM to rats did not show any adverse effects. Efficacy studies showed that the dietary supplementation of this phytase significantly improved growth performance and bone mineralization in broiler chickens and piglets fed P-deficient diets, and increased retention of phosphorus (P) and calcium (Ca), and phytate-P degradation in excreta of broiler chickens in a dose-dependent manner. In conclusion, there are no safety concerns using phytase HM as a feed additive and the phytase is well tolerated by broiler chickens and pigs. Further, phytase HM improves with high efficacy the growth performance in both broiler chickens and pigs., Competing Interests: The authors have affiliated with companies with commercial interest in animal feed but none of the authors has any conflicts of interest concerning this paper., (© 2021 The Author(s).)

Sensory properties of some foods may be of importance to energy consumption and thus the development and maintenance of childhood obesity. This study compares selected food related qualities in overweight and normal weight children. Ninety-two participants were included; 55 were overweight with a mean age of 11.6 years (range 6-18 years) and a mean BMI z-score of 2.71 (range 1.29-4.60). The 37 normal weight children had a mean age of 13.0 years (range 6-19 years) and a mean BMI z-score of 0.16 (range -1.71 to 1.24). All children completed a half-hour long meal test consisting of alternation between consumption of foods and answering of questionnaires. Compared to the normal weight, the overweight children displayed lower self-reported intake paces (χ 2 (2) = 6.3, p = 0.04), higher changes in liking for mozzarella (F(1,63) = 9.55, p = 0.003) and pretzels (F(1,87) = 5.27, p = 0.024), and declines in wanting for something fat, of which the normal weight children displayed an increase (F(1,83) = 4,10, p = 0.046). No differences were found for sensory-specific satiety, wanting for the main food yoghurt, hunger, or satiety. In conclusion, overweight children did not differ from normal weight children in terms of sensory-specific satiety, hunger, or satiety. However, overweight children had lower intake paces and appeared to differ from normal weight children regarding foods with a fatty taste., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Ectopic fat deposition in liver and skeletal muscle tissue is related to cardiovascular disease risk and is a common metabolic complication in obese children. We evaluated the hypotheses of ectopic fat in these organs could be diminished following 1 year of multidisciplinary care specialized in childhood obesity, and whether this reduction would associate with changes in other markers of metabolic function., Methods: This observational longitudinal study evaluated 40 overweight children and adolescents enrolled in a multidisciplinary treatment protocol at the Children's Obesity Clinic, Holbæk, Denmark. The participants were assessed by anthropometry, fasting blood samples (HbA1c, glucose, insulin, lipids, and biochemical variables of liver function), and liver and muscle fat content assessed by magnetic resonance spectroscopy at enrollment and following an average of 12.2 months of care. Univariate linear regression models adjusted for age, sex, treatment duration, baseline degree of obesity, and pubertal developmental stage were used for investigating possible associations., Results: The standard deviation score (SDS) of baseline median body mass index (BMI) was 2.80 (range: 1.49-3.85) and the median age was 14 years (10-17). At the end of the observational period, the 40 children and adolescents (21 girls) significantly decreased their BMI SDS, liver fat, muscle fat, and visceral adipose tissue volume. The prevalence of hepatic steatosis changed from 28 to 20 % (p = 0.26) and the prevalence of muscular steatosis decreased from 75 to 45 % (p = 0.007). Changes in liver and muscle fat were independent of changes in BMI SDS, baseline degree of obesity, duration of treatment, age, sex, and pubertal developmental stage., Conclusions: A 1-year multidisciplinary intervention program in the setting of a childhood obesity outpatient clinic confers a biologically important reduction in liver and muscle fat; metabolic improvements that are independent of the magnitude of concurrent weight loss., Trial Registration: ClinicalTrials.gov registration number: NCT00928473 , the Danish Childhood Obesity Biobank. Registered June 25, 2009.

Introduction: The aim was to investigate whether a familial predisposition to obesity related cardiovascular complications was associated with the degree of obesity at baseline and/or changes in the degree of obesity during a multidisciplinary childhood obesity treatment program., Methods: The study included 1421 obese children (634 boys) with a median age of 11.5 years (range 3.1-17.9 years), enrolled in treatment for 0.04 to 5.90 years (median 1.3 years) at the Children's Obesity Clinic, Denmark. At baseline, weight and height were measured, body mass index (BMI) standard deviation score (SDS) calculated, and self-reported information on familial predisposition to obesity, hypertension, type 2 diabetes mellitus (T2DM), thromboembolic events, and dyslipidaemia were obtained. A familial predisposition included events in biological parents, siblings, grandparents, uncles, and aunts. The treatment outcomes were categorically analysed according to the prevalence of familial predispositions., Results: The median BMI SDS at enrollment was 3.2 in boys and 2.8 in girls. One-thousand-and-forty-one children had obesity in their family, 773 had hypertension, 551 had T2DM, 568 had thromboembolic events, and 583 had dyslipidaemia. Altogether, 733 had three or more predispositions. At baseline, familial T2DM was associated with a higher mean BMI SDS (p = 0.03), but no associations were found between the other predispositions and the children's degree of obesity. During treatment, girls with familial obesity lost more weight, compared to girls without familial obesity (p = 0.04). No other familial predispositions were associated with changes in BMI SDS during treatment., Conclusion: Obese children with a familial predisposition to T2DM showed a significantly higher degree of obesity at baseline and girls with familial obesity responded better to treatment. Besides these findings, no other associations were found between the occurrence of familial predispositions and the degree of obesity or changes herein during multidisciplinary childhood obesity treatment.

The complement system plays a pathophysiological role in systemic lupus erythematosus (SLE). This study aims to investigate whether an association exists between the ficolins that are part of the lectin complement pathway and SLE. EDTA plasma samples from 68 Danish SLE patients and 29 healthy donors were included in the study. Plasma concentrations of Ficolin-1, -2, and -3 were determined in specific sandwich ELISAs. Lectin pathway activity via Ficolin-3 was measured in ELISA on acetylated bovine serum albumin (acBSA) and measured as Ficolin-3 binding and deposition of C4, C3 and the terminal complement complex (TCC). SLE patients had increased levels of Ficolin-3, 21.6μg/ml as compared to 17.0μg/ml in healthy controls (P=0.0098). The Ficolin-1 plasma concentration was negatively correlated with SLE Disease Activity Index (SLEDAI) (Rho=-0.29, P=0.015) and positively correlated to the [Systemic Lupus International Collaborating Clinics (SLICC)/American College of Rheumatology (ACR) Damage Index] (SDI) (Rho=0.27, P=0.026). The Ficolin-1 concentration was also associated with the occurrence of arterial (P=0.0053) but not venous thrombosis (P=0.42). Finally, deposition of C4, C3 and TCC in the Ficolin-3 pathway were all correlated to SLEDAI, respectively (P<0.0076). The Ficolin-1 association to SLEDAI and SDI as well as arterial thrombosis shown in this study suggests that Ficolin-1 may be a potential new biomarker for patients with SLE. Furthermore, Ficolin-3 mediated complement activation may be valuable in monitoring disease activity in SLE patients due to the high sensitivity for complement consumption in the assay independent of the Ficolin-3 concentration., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The prevalence of childhood obesity has reached alarming rates world-wide. The aetiology seems to be an interplay between genetic and environmental factors, and a surrogate measure of this complex interaction is suggested as familial predisposition. Familial predisposition to obesity and related cardiovascular disease (CVD) complications constitute the presence of obesity and/or obesity-related complications in primarily blood-related family members. The approaches of its measurement and applicability vary, and the evidence especially of its influence on obesity and obesity treatment in childhood is limited. Studies have linked a familial predisposition of obesity, CVD (hypertension, dyslipidaemia and thromboembolic events), and type 2 diabetes mellitus to BMI as well as other adiposity measures in children, suggesting degrees of familial aggregation of metabolic derangements. A pattern of predispositions arising from mothers, parents or grandparents as being most influential have been found, but further comprehensive studies are needed in order to specify the exact implications of familial predisposition. In the scope of childhood obesity this article reviews the current literature regarding familial predisposition to obesity and obesity-related complications, and how these familial predispositions may impact obesity in the offspring., (© 2015 S. Karger GmbH, Freiburg.)

Antimicrobial peptides are a new class of antibiotics that are promising for pharmaceutical applications because they have retained efficacy throughout evolution. One class of antimicrobial peptides are the defensins, which have been found in different species. Here we describe a new fungal defensin, eurocin. Eurocin acts against a range of Gram-positive human pathogens but not against Gram-negative bacteria. Eurocin consists of 42 amino acids, forming a cysteine-stabilized α/β-fold. The thermal denaturation data point shows the disulfide bridges being responsible for the stability of the fold. Eurocin does not form pores in cell membranes at physiologically relevant concentrations; it does, however, lead to limited leakage of a fluorophore from small unilamellar vesicles. Eurocin interacts with detergent micelles, and it inhibits the synthesis of cell walls by binding equimolarly to the cell wall precursor lipid II.

NZ2114 is a novel plectasin derivative with potent activity against gram-positive bacteria, including multiply drug-resistant strains. We used the neutropenic murine thigh infection model to characterize the time course of antimicrobial activity of NZ2114 and determine which pharmacokinetic/pharmacodynamic (PK/PD) index and magnitude best correlated with efficacy. Serum drug levels following administration of three fourfold-escalating single-dose levels of NZ2114 were measured by microbiologic assay. Single-dose time-kill studies following doses of 10, 40, and 160 mg/kg of body weight demonstrated concentration-dependent killing over the dose range (0.5 to 3.7 log(10) CFU/thigh) and prolonged postantibiotic effects (3 to 15 h) against both Staphylococcus aureus and Streptococcus pneumoniae. Mice had 10(6.3) to 10(6.8) CFU/thigh of strains of S. pneumoniae or S. aureus at the start of therapy when treated for 24 h with 0.625 to 160 mg/kg/day of NZ2114 fractionated for 4-, 6-, 12-, and 24-h dosing regimens. Nonlinear regression analysis was used to determine which PK/PD index best correlated with microbiologic efficacy. Efficacies of NZ2114 were similar among the dosing intervals (P = 0.99 to 1.0), and regression with the 24-h area under the concentration-time curve (AUC)/MIC index was strong (R(2), 0.90) for both S. aureus and S. pneumoniae. The maximum concentration of drug in serum/MIC index regression was also strong for S. pneumoniae (R(2), 0.96). Studies to identify the PD target for NZ2114 utilized eight S. pneumoniae and six S. aureus isolates and an every-6-h regimen of drug (0.156 to 160 mg/kg/day). Treatment against S. pneumoniae required approximately twofold-less drug for efficacy in relationship to the MIC than did treatment against S. aureus. The free drug 24-h AUCs/MICs necessary to produce a stasis effect were 12.3 +/- 6.7 and 28.5 +/- 11.1 for S. pneumoniae and S. aureus, respectively. The 24-h AUC/MIC associated with a 1-log killing endpoint was only 1.6-fold greater than that needed for stasis. Resistance to other antimicrobial classes did not impact the magnitude of the PD target required for efficacy. The PD target in this model should be considered in the design of clinical trials with this novel antibiotic.

Background: Clinical studies investigating platelet function in dogs have had conflicting results that may be caused by normal physiologic variation in platelet response to agonists., Objectives: The objective of this study was to investigate platelet function in clinically healthy dogs of 4 different breeds by whole-blood aggregometry and with a point-of-care platelet function analyzer (PFA-100), and to evaluate the effect of acetylsalicylic acid (ASA) administration on the results from both methods., Methods: Forty-five clinically healthy dogs (12 Cavalier King Charles Spaniels [CKCS], 12 Cairn Terriers, 10 Boxers, and 11 Labrador Retrievers) were included in the study. Platelet function was assessed by whole-blood aggregation with ADP (1, 5, 10, and 20 microM) as agonist and by PFA-100 using collagen and epinephrine (Col + Epi) and Col + ADP as agonists. Plasma thromboxane B(2) concentration was determined by an enzyme immunoassay. To investigate the effect of ASA, 10 dogs were dosed daily (75 or 250 mg ASA orally) for 4 consecutive days., Results: A higher platelet aggregation response was found in CKCS compared to the other breeds. Longer PFA-100 closure time (Col + Epi) was found in Cairn Terriers compared to Boxers. Plasma thromboxane B(2) concentration was not statistically different between groups. Administration of ASA prolonged the PFA-100 closure times, using Col + Epi (but not Col + ADP) as agonists. Furthermore, ASA resulted in a decrease in whole-blood platelet aggregation., Conclusions: Platelet function is influenced by breed, depending upon the methodology applied. However, the importance of these breed differences remains to be investigated. The PFA-100 method with Col + Epi as agonists, and ADP-induced platelet aggregation appear to be sensitive to ASA in dogs.

Chronic musculoskeletal pain has biological, psychological and social components. This review deals with the biological factors, with emphasis on the fibromyalgia syndrome (FMS). Studies on central sensitization of pain-transmitting neurons, changes in endogenous pain modulation that give rise to pain disinhibition, referred pain, pain-related decrease in muscle strength and endurance, and pain generators in deep tissues are reviewed. In FMS there is strong scientific support for the statement that the biological part of the syndrome is a longstanding or permanent change in the function of the nociceptive nervous system that can be equated with a disease. Further research is necessary in order to determine which methods are best for diagnosis of the pain hypersensitivity in clinical practice. FMS may be the far end of a continuum that starts with chronic localized/regional musculoskeletal pain and ends with widespread chronic disabling pain.

Neural tube defects (NTDs) are congenital malformations of the central nervous system (CNS) secondary to abnormal closure of the neural tube during embryonic development. This study provides information on NTD rate, distribution, associated morphologic anomalies and organ weights in a Danish fetal and perinatal autopsy series during a 16 year period. The data were extracted from the autopsy reports of a consecutive series of 1984 fetal and perinatal autopsies from the Copenhagen area performed in the period 1989-2004. Registered parameters included: The location and morphology of the NTD, associated morphological anomalies, and organ weights. Organ weights were evaluated according to new fetal autopsy standards and grouped as low, normal or high. Ninety-seven NTD cases were found (4.9%): Spina bifida (38 cases), cephalocele (17 cases) and anencephaly (42 cases). 63% of NTD cases had associated morphologic anomalies. Among the most common were hydrocephalus, NTD in another region, and anomalies in the urogenital system. 58% of the NTD cases had abnormal weight of one or more organs. Most notable was low adrenal weight not only in anencephalic fetuses but also in cases with cephalocele, suggesting a possible association.

This paper introduces the design and implementation of a generic wireless and Real-time Multi-purpose Health Care Telemedicine system applying Bluetooth protocol, Global System for Mobile Communications (GSM) and General Packet Radio Service (GPRS). The paper explores the factors that should be considered when evaluating different technologies for application in telemedicine system. The design and implementation of an embedded wireless communication platform utilising Bluetooth protocol is described, and the implementation problems and limitations are investigated. The system is tested and its telecommunication general aspects are verified. The results showed that the system has (97.9 +/- 1.3)% Up-time, 2.5 x 10(-5) Bit Error Rate, 1% Dropped Call Rate, 97.4% Call Success Rate, 5 second transmission delay in average, (3.42 +/- 0.11) kbps throughput, and the system may have application in electrocardiography.

Purpose: The objective of this study was to compare the effect of phosphoric acid and a self-etching adhesive on the short and long-term bond strength of a light-curing sealant to unground primary and permanent enamel., Methods: A light-curing resin sealant (Delton Light Curing Pit & Fissure Sealant-CLEAR) was bonded to the flattest, peripheral surface of 40 primary and 40 permanent molars following conditioning of the cleaned enamel with 38% phosphoric acid or with the self-etching adhesive Prompt L-Pop (N=10/group). After either 1 week or 1 year in water, shear bond strengths were measured. Failure mode was determined in a stereo microscope., Results: There was no significant difference in bond strengths between the phosphoric acid-etch and the self-etching adhesive groups, nor between the 1-week and 1-year results (P>.05). However, the bond strengths to primary enamel were lower than those to permanent enamel (P=.0021). The number of pure adhesive failures in each of the 8 groups varied between 0 and 3 (0-30%), and the remaining teeth displayed mixed adhesive-cohesive failures., Conclusions: The self-etching adhesive studied seems an attractive alternative to the acid-etch technique for sealant application in young children where simplifications in the clinical procedure are warranted.

Introduction: At the Department of Forensic Medicine, University of Aarhus, Denmark, examinations are performed of children who are suspected of having been sexually abused. The medical forensic investigation aims at documenting sexual abuse if there are any physical findings. The presence of certain sexually transmitted diseases, which cannot be explained otherwise, may be such documentation. This article focuses on Chlamydia trachomatis and Neisseria gonorrhoeae, and the risk for the sexually abused child to contract these diseases., Material and Methods: Retrospectively, we went through the Institute's files from 1996 to 2000 concerning child abuse. Information about age, sex, cultures for Chlamydia trachomatis and for Neisseria gonorrhoeae, and the results of these cultures was registered. If the child had been cultured, the suspected perpetrator's age and gender were registered as well. The children were all between 0 and 15 years of age, and in all cases the suspicion of sexual abuse had led to reporting to the police., Results: 295 girls and 41 boys were examined in the period from 1996 to 2000. A total of 100 cultures for Chlamydia trachomatis and 105 cultures for Neisseria gonorrhoeae were performed. Among the 111 children who were examined for Chlamydia trachomatis and/or Neisseria gonorrhoeae there were no positive results. In 102 cases the suspected perpetrator was known to be one or more men, and only in one case it was a women. The mean age of the suspected perpetrators was 35.4 years., Discussion: The prevalence of Neisseria gonorrhoeae and Chlamydia trachomatis should reflect the prevalence in the group of perpetrators. Neisseria gonorrhoeae is uncommon outside Copenhagen, the capital of Denmark, whereas Chlamydia trachomatis is quite common. On this background, we had expected some of the children to be infected with Chlamydia trachomatis. Therefore, the risk for sexually abused children to be infected with Chlamydia trachomatis or Neisseria gonorrhoeae is very low, depending on the kind of abuse and the perpetrator's possible risk behaviour, age, and gender. As a conclusion, only children with a clear indication should be tested for these sexually transmitted diseases.

The in vitro protein-chemical features and the molecular background of osteogenesis imperfecta (OI), a heritable disorder of collagen I metabolism, have been elucidated in recent years. The aim of our study was to find the prevalence of dentinogenesis imperfecta (DI) and other dental anomalies in 88 patients with OI, to compare clinical with radiologic abnormalities, and to correlate these clinical/radiologic findings with the results of gel electrophoresis and molecular studies of collagen I. Twenty-eight percent of OI patients had DI. Most patients with DI had radiologic abnormalities, but some patients had radiologic signs compatible with DI, but no clinical signs of DI. OI type I patients with DI were more severely affected by OI than those without DI. In OI type III and IV, in contrast, there was no difference in overall severity between patients with and without DI. DI was not associated with any particular molecular aberration in any OI type. If defining DI from the presence of both clinical and radiologic signs, collagen I produced by cultured fibroblasts was qualitatively abnormal from all OI patients with DI. Some OI patients had dental abnormalities not resembling DI. A qualitative collagen abnormality could not be found in any of these patients. Denticles, i.e., calcifications within the pulpal cavity, were found more frequently in OI patients than in control subjects.

Urinary pyridinoline (Pyd) and deoxypyridinoline (Dpd) are markers of bone resorption that are elevated above normal in subjects with metabolic bone disease. Total Pyd and Dpd, both free and peptide-bound forms, can be measured by HPLC after hydrolysis and cellulose chromatography. Since free Pyd is the major component of total Pyd in urine, we developed an immunoassay using free Pyd as an immunogen. This assay is much easier to perform than HPLC, requires no sample preparation, and correlates well with total Pyd measurement by HPLC (r = 0.97) and with urinary hydroxyproline (r = 0.90). The antiserum reacts most strongly with free Pyd and Dpd and minimally with glycosylated and large peptide-bound forms. The sensitivity of the Pyd immunoassay is less than 25 nM. The intraassay CV is 5-10%; the interassay CV is 10-15%. Analytic recovery studies indicated negligible sample interference. Furthermore, measurement of the Pyd in the same individuals over a 30 day time period exhibited minimal day-to-day variation. Thus, the Pyd immunoassay provides a rapid and easy method for evaluation of Pyd in urine. Pyd immunoassay may serve as a practical method of screening for metabolic bone disease and for monitoring therapeutic treatment.

This study examines the influence of sagittal occlusion on occlusal plaque formation in permanent first molars (PFM) in 72 7-10-yr-olds before loss of primary second molars. Of a total of 288 PFM, 140 (49%) were sealed and 23 (8%) filled. Occlusal plaque was recorded at two levels of examination: 1) visible plaque on the entire surface and 2) detailed macromorphologic mapping. Enamel caries was recorded after professional tooth cleaning. After 48 h without tooth brushing, plaque examinations were repeated. Stone models were used for 1) identification of interocclusal contact areas and 2) classification of sagittal molar occlusion. The detailed mapping of plaque on unfilled surfaces showed a clear pattern of preferential locations related to the macromorphology of the occlusal surfaces. Active caries was restricted to those anatomic structures where plaque accumulated. 48-h median plaque values on mandibular molars in normal and with one cusp distal occlusion were significantly lower (P < 0.01) compared to surfaces in 1/4, 1/2 and 3/4 distal molar occlusion. Maxillary molars with normal and with 1 cusp distal occlusion had lower median plaque values than other sagittal occlusion categories. In general, however, plaque scores were higher in maxillary teeth because more than 2/3 of these teeth were without occlusal contact in the distal part. Mandibular occlusal surfaces in normal and with one cusp distal occlusion had significantly fewer active lesions than teeth with 1/4, 1/2 and 3/4 cusp distal occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)

During the past decade usage of the fissure sealant technique for occlusal caries prevention has been increasingly recommended. This study explores variations in indications and usage of this technique in Denmark. A questionnaire was sent to 205 chief dental officers in the Danish Public Child Dental Health Service (PDHS) covering 90% of Danish children and to municipalities, where dental health to children is provided by private practitioners. More than half of the respondents did not use firmly defined criteria for fissure sealant application. Oral hygiene and previous caries experience were most often stated as indications. Actual usage of the sealant technique differed significantly from the PDHS and private clinics. Thus, 33% of the chief dental officers in the PDHS said that sealants were routinely applied to 8- and 13-year-olds, 43% to 30-80% of the children, and 15% used sealants to less than 10% of 8- and 13-year-olds. In contrast, only 5% of the private clinics reported routinely use of sealing technique, 22% used application to 30-80% of the children, while more than one third of the private practitioners used sealants to less than 10% of the 8- and 13-year-olds. In spite of the significant difference in sealant usage between PDHS and private practitioners, it was not possible to see a corresponding difference neither in caries prevalence nor in occlusal filling incidence. Moreover, analysis of PDHS caries prevalence data revealed that PDHS variations could not be explained by variations of risk in terms of social classes and caries incidence.(ABSTRACT TRUNCATED AT 250 WORDS)

Unlabelled: A total of 105 children with cerebral palsy aged 14 and 15 years born in the eastern Denmark comprised the study group. The children were classified according to CP-diagnosis, mental handicap, motor handicap and information on plaque index, gingivitis index, and toothbrushing habits were collected. The analysis showed that the values for plaque and gingivitis indices were significantly higher than those of the control group. The specific CP-diagnosis, the mental handicap and the motor handicap did not seem to have definite influence on the child's plaque and gingivitis indices. Children with the mildest mental and motor handicap seemed to have lower plaque and gingivitis indices than those of the severely handicapped children. The highest gingival scores (5.2) were seen when the child and the parents together were responsible for the child's toothbrushing. The gingival index was 4.0, when the parents brushed the child's teeth and 3.7 when the children brushed their teeth on their own. Only 7 children used an electric toothbrush. The average gingival index in this group was 3.0, which was nearly the same as that of the control group., Conclusion: 1. Oral hygiene planning of the CP-child can only be made when an individual evaluation has been performed. 2. CP-children's oral hygiene has to be controlled frequently.

A total of 105 children with cerebral palsy, born in eastern Denmark in 1968 or 1969, comprised the group studied here. They were classified by CP-diagnosis, motor handicap, mental handicap, speech, caries, with dental data statistically analyzed. The caries rate of the combined CP-group was significantly lower than that of the control group.

Three cases of intrusion of primary teeth are described. In all cases the intruded primary tooth was left without treatment and the tooth did not re-erupt. After eruption of the succedaneous permanent tooth the primary tooth was surgically removed. There was damage to the permanent tooth in two of the three cases.