The infant gut virome is associated with preschool asthma risk independently of bacteria.

Bacteriophage (also known as phage) communities that inhabit the gut have a major effect on the structure and functioning of bacterial populations, but their roles and association with health and disease in early life remain unknown. Here, we analyze the gut virome of 647 children aged 1 year from the Copenhagen Prospective Studies on Asthma in Childhood2010 (COPSAC2010) mother-child cohort, all deeply phenotyped from birth and with longitudinally assessed asthma diagnoses. Specific temperate gut phage taxa were found to be associated with later development of asthma. In particular, the joint abundances of 19 caudoviral families were found to significantly contribute to this association. Combining the asthma-associated virome and bacteriome signatures had additive effects on asthma risk, implying an independent virome-asthma association. Moreover, the virome-associated asthma risk was modulated by the host TLR9 rs187084 gene variant, suggesting a direct interaction between phages and the host immune system. Further studies will elucidate whether phages, alongside bacteria and host genetics, can be used as preclinical biomarkers for asthma.

The airway microbiota of neonates colonized with asthma-associated pathogenic bacteria.

Culture techniques have associated colonization with pathogenic bacteria in the airways of neonates with later risk of childhood asthma, whereas more recent studies utilizing sequencing techniques have shown the same phenomenon with specific anaerobic taxa. Here, we analyze nasopharyngeal swabs from 1 month neonates in the COPSAC2000 prospective birth cohort by 16S rRNA gene sequencing of the V3-V4 region in relation to asthma risk throughout childhood. Results are compared with previous culture results from hypopharyngeal aspirates from the same cohort and with hypopharyngeal sequencing data from the later COPSAC2010 cohort. Nasopharyngeal relative abundance values of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are associated with the same species in the hypopharyngeal cultures. A combined pathogen score of these bacteria's abundance values is associated with persistent wheeze/asthma by age 7. No other taxa are associated. Compared to the hypopharyngeal aspirates from the COPSAC2010 cohort, the anaerobes Veillonella and Prevotella, which have previously been implicated in asthma development, are less commonly detected in the COPSAC2000 nasopharyngeal samples, but correlate with the pathogen score, hinting at latent community structures that bridge current and previous results. These findings have implications for future asthma prevention efforts.

Marker for kidney fibrosis is associated with inflammation and deterioration of kidney function in people with type 2 diabetes and microalbuminuria.

BACKGROUND: Diabetic kidney disease is a major cause of morbidity and mortality. Dysregulated turnover of collagen type III is associated with development of kidney fibrosis. We investigated whether a degradation product of collagen type III (C3M) was a risk marker for progression of chronic kidney disease (CKD), occurrence of cardiovascular disease (CVD), and mortality during follow up in people with type 2 diabetes (T2D) and microalbuminuria. Moreover, we investigated whether C3M was correlated with markers of inflammation and endothelial dysfunction at baseline. METHODS: C3M was measured in serum (sC3M) and urine (uC3M) in 200 participants with T2D and microalbuminuria included in an observational, prospective study at Steno Diabetes Center Copenhagen in Denmark from 2007-2008. Baseline measurements included 12 markers of inflammation and endothelial dysfunction. The endpoints were CVD, mortality, and CKD progression (>30% decline in eGFR). RESULTS: Mean (SD) age was 59 (9) years, eGFR 90 (17) ml/min/1.73m2 and median (IQR) urine albumin excretion rate 102 (39-229) mg/24-h. At baseline all markers for inflammation were positively correlated with sC3M (p≤0.034). Some, but not all, markers for endothelial dysfunction were correlated with C3M. Median follow-up ranged from 4.9 to 6.3 years. Higher sC3M was associated with CKD progression (with mortality as competing risk) with a hazard ratio (per doubling) of 2.98 (95% CI: 1.41-6.26; p = 0.004) adjusted for traditional risk factors. uC3M was not associated with CKD progression. Neither sC3M or uC3M were associated with risk of CVD or mortality. CONCLUSIONS: Higher sC3M was a risk factor for chronic kidney disease progression and was correlated with markers of inflammation.

The developing airway and gut microbiota in early life is influenced by age of older siblings.

BACKGROUND: Growing up with siblings has been linked to numerous health outcomes and is also an important determinant for the developing microbiota. Nonetheless, research into the role of having siblings on the developing microbiota has mainly been incidental. RESULTS: Here, we investigate the specific effects of having siblings on the developing airway and gut microbiota using a total of 4497 hypopharyngeal and fecal samples taken from 686 children in the COPSAC2010 cohort, starting at 1 week of age and continuing until 6 years of age. Sibship was evaluated longitudinally and used for stratification. Microbiota composition was assessed using 16S rRNA gene amplicon sequencing of the variable V4 region. We found siblings in the home to be one of the most important determinants of the developing microbiota in both the airway and gut, with significant differences in alpha diversity, beta diversity, and relative abundances of the most abundant taxa, with the specific associations being particularly apparent during the first year of life. The age gap to the closest older sibling was more important than the number of older siblings. The signature of having siblings in the gut microbiota at 1 year was associated with protection against asthma at 6 years of age, while no associations were found for allergy. CONCLUSIONS: Having siblings is one of the most important factors influencing a child's developing microbiota, and the specific effects may explain previously established associations between siblings and asthma and infectious diseases. As such, siblings should be considered in all studies involving the developing microbiota, with emphasis on the age gap to the closest older sibling rather than the number of siblings. Video abstract.

Prevention of EAE by tolerogenic vaccination with PEGylated antigenic peptides.

BACKGROUND: Therapeutic treatment options for chronic autoimmune disorders such as multiple sclerosis (MS) rely largely on the use of non-specific immunosuppressive drugs, which are not able to cure the disease. Presently, approaches to induce antigen-specific tolerance as a therapeutic approach; for example, by peptide-based tolerogenic 'inverse' vaccines have regained great interest. We have previously shown that coupling of peptides to carriers can enhance their capacity to induce regulatory T cells in vivo. METHOD: In this present study, we investigated whether the tolerogenic potential of immunodominant myelin T-cell epitopes can be improved by conjugation to the synthetic carrier polyethylene glycol (PEG) in an experimental autoimmune encephalomyelitis (EAE) mouse model for chronic MS (MOG C57BL/6). RESULTS: Preventive administration of the PEGylated antigenic peptide could strongly suppress the development of EAE, accompanied by reduced immune cell infiltration in the central nervous system (CNS). Depletion of regulatory T cells (Tregs) abrogated the protective effect indicating that Tregs play a crucial role in induction of antigen-specific tolerance in EAE. Treatment during the acute phase of disease was safe and did not induce immune activation. However, treatment at the peak of disease did not affect the disease course, suggesting that either induction of Tregs does not occur in the highly inflamed situation, or that the immune system is refractory to regulation in this condition. CONCLUSION: PEGylation of antigenic peptides is an effective and feasible strategy to improve tolerogenic (Treg-inducing) peptide-based vaccines, but application for immunotherapy of overt disease might require modifications or combination therapies that simultaneously suppress effector mechanisms.

Long-Term Outcome of Patients Operated with Pars Plana Vitrectomy for Primary Rhegmatogenous Retinal Detachment.

PURPOSE: To determine the long-term outcome of patients operated with pars plana vitrectomy (PPV) for primary rhegmatogenous retinal detachment (RRD) and to identify potential predictors for poor visual outcome. METHODS: Prospective, observational 30-month study of patients operated for primary RRD with PPV. Examinations were performed preoperatively and after months 2, 6, and 30. RESULTS: Eighty-four patients (84 eyes) were included and 73 (86.9%) participated at month 30. The macula was attached in 30 (35.7%) patients at primary operation. The majority of patients (n = 59, 80.8%) achieved a good final best corrected visual acuity (BCVA ≤0.3 logMAR, ≥0.5 Snellen) with a better outcome in patients with the macula attached than detached (0.02 vs. 0.17 logMAR, p = 0.007). Variables associated with poor visual outcome were baseline BCVA >0.3 logMAR (p = 0.03), female gender (p = 0.02), silicone oil (p = 0.03), and larger areas of retinal detachment (p = 0.01). In multivariable regression analysis, female gender (OR = 8.5 [95% CI 1.8-39.8]) was the strongest risk factor for poor visual outcome. CONCLUSION: The majority of patients operated for primary RRD achieved a reasonable long-term visual outcome. Notably, female gender was associated with poor visual outcome, indicating a need for closer follow-up.

Fundus autofluorescence and spectral domain optical coherence tomography as predictors for long-term functional outcome in rhegmatogenous retinal detachment.

PURPOSE: To detect pre- and postoperative retinal changes in fundus autofluorescence (AF) and spectral domain optical coherence tomography (SD-OCT) and to correlate these with functional outcome in patients with primary rhegmatogenous retinal detachment (RRD). METHODS: A prospective, 30-month study of patients operated with 25-gauge vitrectomy for primary RRD. Patients were examined preoperatively and after 6 and 30 months, using ultrawide-field AF images (UWFI) (Optos 200Tx) and SD-OCT (Topcon 3D OCT-2000) imaging. RESULTS: Of 84 patients (84 eyes) included at baseline, 100.0 and 86.9% were re-examined at month 6 and 30, respectively. Preoperative findings such as macular attachment, detachment > 750 µm from foveola, lack of intraretinal separation, and subfoveal elevation ≤ 500 µm were all associated with better BCVA at months 6 and 30. Postoperative disruption of the photoreceptor layer was associated with poor BCVA at month 6 (p < 0.001) but not at month 30. At baseline, AF-demarcation of RRD was demonstrated by a hyperfluorescent edge in 92.0% and was associated with visual impairment at months 6 (p = 0.003) and 30 (p = 0.003). Visual outcome at month 30 was good (≤ 0.3 logMAR (≥ 20/40 Snellen)), regardless of the preoperative, macular status. However, with significantly better visual outcome in patients with macula attachments versus partly or totally macular detachments (p < 0.001). CONCLUSION: Fundus AF and SD-OCT is able to identify retinal reestablishment up to 30 months after primary RRD, with good correlation to BCVA. These findings emphasize the importance of long-term studies for final visual recovery.

Possible adverse effects of the quadrivalent human papillomavirus vaccine in the Region of Southern Denmark: a retrospective, descriptive cohort study.

INTRODUCTION: Since the introduction of the quadrivalent human papillomavirus vaccine, young girls and women have reported a broad range of symptoms. These have been described as possible adverse effects of the vaccine. In this study, we describe demographic characteristics, symptomatology, clinical and laboratory test results in patients referred with suspected adverse effects in the Region of Southern Denmark. METHODS: We conducted a retrospective, descriptive study. The patients filled out a questionnaire, were interviewed by a doctor and received a standard physical examination and laboratory tests. RESULTS: The study comprised 200 patients. The median age at referral was 22 (interquartile range (IQR): 19.5-26) years, and age at first vaccination was 14 (IQR: 12-21) years. The most common symptoms were headache (93%), fatigue/tiredness (93%) and dizziness when standing up (90%). The median number of symptoms in each patient was 15. Five patients (2.5%) were diagnosed with postural orthostatic tachycardia syndrome (POTS). Of all patients, 183 (91.5%) were terminated without a somatic diagnosis, one patient (0.5%) was terminated with a functional disorder and 11 patients (5.5%) were still in diagnostic workup when the present study concluded. CONCLUSIONS: The patients reported a wide range of symptoms. We found an overall low prevalence of POTS. It should be further investigated whether these patients might suffer from a functional disorder rather than from adverse effects associated with the vaccine. FUNDING: none. TRIAL REGISTRATION: not relevant.

Noninvasive Retinal Markers in Diabetic Retinopathy: Advancing from Bench towards Bedside.

The retinal vascular system is the only part of the human body available for direct, in vivo inspection. Noninvasive retinal markers are important to identity patients in risk of sight-threatening diabetic retinopathy. Studies have correlated structural features like retinal vascular caliber and fractals with micro- and macrovascular dysfunction in diabetes. Likewise, the retinal metabolism can be evaluated by retinal oximetry, and higher retinal venular oxygen saturation has been demonstrated in patients with diabetic retinopathy. So far, most studies have been cross-sectional, but these can only disclose associations and are not able to separate cause from effect or to establish the predictive value of retinal vascular dysfunction with respect to long-term complications. Likewise, retinal markers have not been investigated as markers of treatment outcome in patients with proliferative diabetic retinopathy and diabetic macular edema. The Department of Ophthalmology at Odense University Hospital, Denmark, has a strong tradition of studying the retinal microvasculature in diabetic retinopathy. In the present paper, we demonstrate the importance of the retinal vasculature not only as predictors of long-term microvasculopathy but also as markers of treatment outcome in sight-threatening diabetic retinopathy in well-established population-based cohorts of patients with diabetes.

[Successful treatment of severe dabigatran intoxication with idarucizumab in a patient with acute kidney injury].

In the course of an uncomplicated sigmoidostomy a 63-year-old male who had severe comorbidity developed a critical bleeding due to dabigatran intoxication induced by acute kidney injury. Massive blood transfusions, tranexamic acid, Octaplex and haemodialysis were not effective. Administration of idarucizumab induced immediate clinical and paraclinical improvement. Dabigatran should be carefully administrated in patients who have any degree of renal insufficiency. Idarucizumab may be effective in severe bleeding caused by dabigatran.

Quality of life development during initial hemodialysis therapy and association with loss of residual renal function.

INTRODUCTION: Health related quality of life (HRQOL) is markedly reduced in hemodialysis patients compared to the general population. We investigated the course of self-reported HRQOL over time and the association with selected factors, focusing on changes in glomerular filtration rate (GFR). METHODS: Eighty-two newly started hemodialysis patients from the SAFIR cohort filled out the Kidney Disease Quality of Life Short Form Version 1.3 (KDQOL-SFTM ) questionnaire at baseline, 6 and 12 months. The SAFIR study was a randomized, placebo-controlled, double-blind intervention study, examining the effects of the angiotensin II receptor blocker irbesartan. HRQOL was a secondary outcome measure. Main inclusion criteria: Dialysis vintage <1 year, left ventricular ejection fraction >30% and urinary output >300 mL/day. GFR was measured with mean creatinine and urea clearance from 24-hour urine collections at baseline, 6 and 12 months. FINDINGS: Irbesartan treatment did not affect HRQOL. Patients were pooled into one group for further analyses. Decline in GFR correlated significantly with decreasing HRQOL over time. HRQOL was stable over time, with a slight nonsignificant tendency toward improved HRQOL. The largest HRQOL-differences (positive values equal improved HRQOL) observed during the 12 month study period were (mean[95% confidence interval]): Burden of kidney disease:6.4[-2.2;15.0], Role limitations-physical:12.7[-2.1;27.5], and Role limitations-emotional:9.7[-5.2;24.6]. Comorbidity, especially diabetes, hospital admissions, female gender, and age were strongly associated with lower HRQOL in cross sectional analysis. DISCUSSION: Preservation of residual renal function seems to be important for HRQOL. In newly started HD patients, HRQOL showed little change after 12 months. HRQOL was negatively affected by comorbidity, especially diabetes, hospital admissions, female gender, and age.

Epidemiologic characteristics of retinal detachment surgery at a specialized unit in Denmark.

PURPOSE: To examine the incidence of retinal detachments and to evaluate patient profiles and surgical characteristics. METHODS: Retrospective review of patients operated for primary retinal detachment (RD) and redetachment between 2010 and 2012 at the Department of Ophthalmology, Odense University Hospital, Denmark. We included all RD such as rhegmatogenous retinal detachment (RRD), tractional retinal detachment (TRD) and exudative retinal detachment (ERD). RESULTS: In total, 779 RD surgeries were performed: 83.7% (n = 652) primary operations and 16.3% (n = 127) reoperation. For primary operation, pars plana vitrectomy (PPV), scleral buckling and combined operations were performed in 95.1% (n = 620), 4.6% (n = 30) and 0.3% (n = 2) respectively. Over time there was less use of silicone oil and greater use of gas tamponade (p = <0.001), less simultaneous cataract operations (p = <0.001), less use of cryotherapy (p = 0.045) and more use of peeling procedures (p = <0.001) in primary operations. The annual incidence of surgery for primary RD was 22.0 [95% confidence interval (CI) 20.4-23.8] per 100 000 inhabitants aged >15 years. Retinal detachment (RD) was more common in males than females (1.8:1), and mean age at presentation was 61.8 years (standard deviations ± 12.3). The annual incidence for RRD and TRD operation was 20.72 (95% CI 19.11-22.43) and 1.25 (95% CI 0.9-1.7) per 100 000 inhabitants >15 years. CONCLUSION: This study presents the first overall incidence for RD in Denmark. The highest incidence of RRD was among males aged 60 to 79 years, whereas TRD was among females at same age. Pars plana vitrectomy (PPV) was the preferred surgical technique, and during 2012 all RD patients were treated with PPV, independent of lens status and age.

Tolerogenic modulation of the immune response by oligoglycerol- and polyglycerol-peptide conjugates.

Peptide-based therapy is a promising strategy for antigen-specific immunosuppression to treat or even heal autoimmune diseases with significantly reduced adverse effects compared to conventional therapies. However, there has been no major success due to the drawbacks of native peptides, i.e., limited bioavailability. Considering the importance and limitations of peptide-based therapies for treatment of autoimmune diseases, we designed and constructed oligoglycerol (OG)- and polyglycerol (PG)-based peptide conjugates. They were evaluated for their biological activity (in vitro and in vivo), bioavailability, and tolerogenic potential. Among the OG- and PG-peptide constructs, PG-peptide constructs exhibited an extended bioavailability compared to OG-peptide constructs and unconjugated peptide. Interestingly, size, structure, and linker chemistry played a critical role for the tolerogenic capacity of the constructs. The PG-peptide construct bound via an ester linkage was the most tolerogenic conjugate, while the PG-peptide construct bound via an amide induced stronger proliferation, but also higher TNF production and lower frequencies of Foxp3(+) regulatory T-cells. Therefore, we conclude that PG-peptide conjugates bound via an ester linkage are not only promising candidates for tolerogenic vaccination, but also open a new avenue toward the application of peptides for the treatment of autoimmune diseases.