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Purpose: In this study, we aimed to determine the effect of a probiotic dietary supplement on physical-anthropometric and biochemical markers in Slovenian patients with metabolic syndrome. Methods: This prospective, randomized, double-blind, placebo-controlled pilot clinical trial involved 16 volunteers who consumed a probiotic dietary supplement over a period of 3 months. The volunteers underwent clinical assessments before and at the end of the period of consuming the dietary supplement or placebo. Results: At a concentration of 1 × 109 colony-forming units per day (N = 8), the Lactobacillus plantarum PCS 26 supplement decreased waist circumference (p = 0.043), lowered serum cholesterol (p = 0.028), and maintained serum adiponectin levels (p = 0.629). In the group that received the placebo (N = 8), no differences were found in the waist circumference (p = 0.750) or serum cholesterol levels (p = 0.225); however, we found a statistically significant reduction of the serum adiponectin levels (p = 0.036). Conclusion: Although the number of patients examined was too low to draw any firm conclusions, dietary supplementation with the probiotic strain Lactobacillus plantarum PCS 26 exerted some favorable effects that could alleviate the symptoms of metabolic syndrome and reduce the clinical manifestations and risks of cardiovascular diseases. These results represent a starting point to carry out an extensive long-term clinical trial that would provide insight into the mechanisms of action of probiotic bacteria in preventing or relieving metabolic syndrome or its components.

BACKGROUND: Uncontrolled pediatric asthma has a large impact on patients and their caregivers. More insight into determinants of uncontrolled asthma is needed. We aim to compare treatment regimens, inhaler techniques, medication adherence and other characteristics of children with controlled and uncontrolled asthma in the: Systems Pharmacology approach to uncontrolled Paediatric Asthma (SysPharmPediA) study. MATERIAL AND METHODS: 145 children with moderate to severe doctor-diagnosed asthma (91 uncontrolled and 54 controlled) aged 6-17 years were enrolled in this multicountry, (Germany, Slovenia, Spain, and the Netherlands) observational, case-control study. The definition of uncontrolled asthma was based on asthma symptoms and/or exacerbations in the past year. Patient-reported adherence and clinician-reported medication use were assessed, as well as lung function and inhalation technique. A logistic regression model was fitted to assess determinants of uncontrolled pediatric asthma. RESULTS: Children in higher asthma treatment steps had a higher risk of uncontrolled asthma (OR (95%CI): 3.30 (1.56-7.19)). The risk of uncontrolled asthma was associated with a larger change in FEV1% predicted post and pre-salbutamol (OR (95%CI): 1.08 (1.02-1.15)). Adherence and inhaler techniques were not associated with risk of uncontrolled asthma in this population. CONCLUSION: This study showed that children with uncontrolled moderate-to-severe asthma were treated in higher treatment steps compared to their controlled peers, but still showed a higher reversibility response to salbutamol. Self-reported adherence and inhaler technique scores did not differ between controlled and uncontrolled asthmatic children. Other determinants, such as environmental factors and differences in biological profiles, may influence the risk of uncontrolled asthma in this moderate to severe asthmatic population. The SysPharmPediA consortium is supported by ZonMW [project number: 9,003,035,001], the Ministry of Education, Science, and Sport of the Republic of Slovenia [contract number C330–16–500,106]; the German Ministry of Education and Research (BMBF) [project number FKZ 031L0088]; Instituto de Salud Carlos III (ISCIII) through Strategic Action for Health Research (AES) and European Community (EC) within the Active and Assisted Living (AAL) Program framework [award numbers AC15/00,015 and AC15/00,058] under the frame of the ERACoSysMed JTC-1 Call.”

Anti-TNF therapy has significantly improved disease control in rheumatoid arthritis, but a fraction of rheumatoid arthritis patients do not respond to anti-TNF therapy or lose response over time. Moreover, the mechanisms underlying non-response to anti-TNF therapy remain largely unknown. To date, many single biomarkers of response to anti-TNF therapy have been published but they have not yet been analyzed as a system of interacting nodes. The aim of our study is to systematically elucidate the biological processes underlying non-response to anti-TNF therapy in rheumatoid arthritis using the gene ontologies of previously published predictive biomarkers. Gene networks were constructed based on published biomarkers and then enriched gene ontology terms were elucidated in subgroups using gene ontology software tools. Our results highlight the novel role of proteasome-mediated protein catabolic processes (p = 2.91 × 10−15) and plasma lipoproteins (p = 4.55 × 10−11) in anti-TNF therapy response. The results of our gene ontology analysis help elucidate the biological processes underlying non-response to anti-TNF therapy in rheumatoid arthritis and encourage further study of the highlighted processes.

Objectives Acute kidney injury (AKI) is associated with higher perioperative mortality and morbidity. Oxidative stress has been proposed as a cause of postoperative AKI. Ascorbic acid (AA) supplementation was suggested as a novel and promising antioxidant. The aim of this study was to evaluate the capability of AA to reduce the incidence of postoperative AKI in cardiac surgery patients. Methods A prospective randomized trial was conducted in patients scheduled for on-pump cardiac surgery. Subjects in the AA group received 2 g of AA intravenously during the induction of anesthesia, 2 g before aortic cross-clamp removal and 1 g every 8 hours for five postoperative days (the JERICA protocol). Postoperatively, the patients were monitored for AKI and other complications. Malondialdehyde levels were monitored in a subpopulation of 100 patients to evaluate the effect of AA on oxidative stress level. Results The AA and control group consisted of 163 and 169 patients, respectively. The groups were well matched for baseline demographics and had similar intraoperative characteristics. The incidence of AKI in the AA and control group was 20.9 and 28.4%, respectively (p = 0.127). The estimated glomerular filtration rate did not differ between the study groups in the entire postoperative period. There was a trend toward higher malondialdehyde values with statistical significance on postoperative day 1 and lower in-hospital mortality in the AA group (0.6 vs. 4.1%, p = 0.067). Conclusion Our results do not support the effectiveness of AA supplementation in reducing the incidence of postoperative AKI in on-pump cardiac surgery patients. Clinical Registration Number This study was registered with the ISRCTN Registry under the trial registration number ISRCTN98572043.

Background: In the last few decades, different coma scoring scales have been proposed. The purpose of this study is to compare two coma scales, the GCS (Glasgow Coma Scale) and the FOUR score (Full Outline of UnResponsiveness score), aiming to examine which scale is better for predicting outcome in traumatic brain injury (TBI) patients in the prehospital setting.Methods: We evaluated the GCS and FOUR scores in the prehospital setting at three different prehospital timepoints, and we reassessed the scores in surviving patients 24 hours, one month and three months after the injury. Then, we compared the outcomes.We used the χ² method, and based on the analysis with the best cut-off point for each model, we calculated the sensitivity, specificity and correct prediction of outcomes with four severity scores. The Youden index, Z score, McNemar's test and ROC curve were also assessed. P < 0.05 was considered statistically significant. Both scales were ranked with gain ratios.Results: We included 200 TBI patients who were treated in a prehospital setting by a prehospital specialized medical unit. In terms of the predictions of positive outcomes, our study showed the following:1. Twenty-four hours after the injury, the best cut-off points obtained were from the FOUR 1 and GCS 1 models.2. One month after the injury, the best cut-off points obtained were from the following models: FOUR 2 and GCS 3 as well as FOUR 3 and GCS 3.3. Three months after the injury, the best cut-off points obtained were from the following models: FOUR 2 and GCS 1, FOUR 3 and GCS 1, and GCS 1 and GCS 2.Conclusions: In our study, the results of our research confirm that there are no practical or clinical differences between the GCS and FOUR scores in terms of predicting morality outcomes 24 hours, one month, and three months after injury. No statistically significant differences were found in the Youden index or the area under the ROC curve 24 hours, one month or three months after the injury.

Autism spectrum disorders (ASD) represent a phenotypically heterogeneous group of patients that strongly intertwine with other neurodevelopmental disorders (NDDs), with genetics playing a significant role in their etiology. Whole exome sequencing (WES) has become predominant in molecular diagnostics for ASD by considerably increasing the diagnostic yield. However, the proportion of undiagnosed patients still remains high due to complex clinical presentation, reduced penetrance, and lack of segregation analysis or clinical information. Thus, reverse phenotyping, where we first identified a possible genetic cause and then determine its clinical relevance, has been shown to be a more efficient approach. WES was performed on 147 Slovenian pediatric patients with suspected ASD. Data analysis was focused on identifying ultrarare or “single event” variants in ASD-associated genes and further expanded to NDD-associated genes. Protein function and gene prioritization were performed on detected clinically relevant variants to determine their role in ASD etiology and phenotype. Reverse phenotyping revealed a pathogenic or likely pathogenic variant in ASD-associated genes in 20.4% of patients, with subsequent segregation analysis indicating that 14 were de novo variants and 1 was presumed compound heterozygous. The diagnostic yield was further increased by 2.7% by the analysis of ultrarare or “single event” variants in all NDD-associated genes. Protein function analysis established that genes in which variants of unknown significance (VUS) were detected were predominantly the cause of intellectual disability (ID), and in most cases, features of ASD as well. Using such an approach, variants in rarely described ASD-associated genes, such as SIN3B, NR4A2, and GRIA1, were detected. By expanding the analysis to include functionally similar NDD genes, variants in KCNK9, GNE, and other genes were identified. These would probably have been missed by classic genotype–phenotype analysis. Our study thus demonstrates that in patients with ASD, analysis of ultrarare or “single event” variants obtained using WES with the inclusion of functionally similar genes and reverse phenotyping obtained a higher diagnostic yield despite limited clinical data. The present study also demonstrates that most of the causative genes in our cohort were involved in the syndromic form of ASD and confirms their comorbidity with other developmental disorders.

Although human clinical studies have suggested probiotic effects on blood glucose levels, knowledge about molecular mechanisms is still scarce. To test the hypothesis that selected Lactobacillus probiotic bacteria could regulate the activity of enterocyte glucose transporters, we aimed to measure in vitro effects of selected Lactobacillus probiotic bacteria on transcription and translation of intestinal glucose transporters sodium-dependent glucose cotransporter 1 (SGLT1) and glucose transporter 2 (GLUT2) as well as transepithelial glucose transport. Lactobacillus plantarum strains (PCS20 and PCS26), Lactobacillus rhamnosus GG (LGG) (ATCC 53103) and Lactobacillus acidophilus (L acidophilus) (ATCC 4356) were co-cultivated with noncarcinogenic porcine enterocytes (CLAB) and human epithelial colorectal adenocarcinoma cells (Caco-2) (ATCC HTB-37). Changes in transcription and expression of SGLT1 and GLUT2 were strain and cell line-specific. In CLAB, LGG was the most potent SGLT1 up-regulator, and PCS26 the most potent down-regulator of GLUT2 transcription, which was also reflected on the protein level. In Caco-2, all tested strains tended to downregulate GLUT2 gene expression, while L acidophilus most effectively reduced GLUT2 protein levels. Statistically significant effect of PCS26 and L acidophilus on GLUT2 molecular and protein levels in CLAB and Caco-2 cell lines, respectively, was also followed by a decreased rate of transepithelial glucose transport. Careful selection of specific Lactobacillus probiotic strains could be used to downregulate glucose absorption in intestinal epithelial cells and thereby could be beneficial as a support treatment of pathologies related to glucose homeostasis.

Crohn’s disease (CD), rheumatoid arthritis, psoriatic arthritis and other inflammatory diseases comprise a group of chronic diseases with immune-mediated pathogenesis which share common pathological pathways, as well as treatment strategies including anti-TNF biologic therapy. However, the response rate to anti-TNF therapy among those diseases varies, and approximately one third of patients do not respond. Since pharmacogenetic studies for anti-TNF therapy have been more frequent for other related diseases and are rare in CD, the aim of our study was to further explore markers associated with anti-TNF response in other inflammatory diseases in Slovenian CD patients treated with the anti-TNF drug adalimumab (ADA). We enrolled 102 CD patients on ADA, for which the response was defined after 4, 12, 20 and 30 weeks of treatment, using an IBDQ questionnaire and blood CRP value. We genotyped 41 SNPs significantly associated with response to anti-TNF treatment in other diseases. We found novel pharmacogenetic association between SNP rs755622 in the gene MIF (macrophage migration inhibitory factor) and SNP rs3740691 in the gene ARFGAP2 in CD patients treated with ADA. The strongest and most consistent association with treatment response was found for the variant rs2275913 in gene IL17A (p = 9.73 × 10−3).

The need for temperature modulation (mostly cooling) in critically ill patients is based on the expected benefits associated with decreased metabolic demands. However, evidence-based guidelines for temperature management in a majority of critically ill patients with fever are still lacking. The aim of our retrospective single-site observational study was to determine the differences in ICU treatment between patients in whom their temperature remained within the target temperature range for ≥25% of time (inTT group) and patients in whom their temperature was outside the target temperature range for

Background Women with uterine adenomyosis seeking assisted reproduction have been associated with compromised endometrial receptivity to embryo implantation. To understand the mechanisms involved in this process, we aimed to compare endometrial transcriptome profiles during the window of implantation (WOI) between women with and without adenomyosis. Methods We obtained endometrial biopsies LH-timed to the WOI from women with sonographic features of adenomyosis (n=10) and controls (n=10). Isolated RNA samples were subjected to RNA sequencing (RNA-seq) by the Illumina NovaSeq 6000 platform and endometrial receptivity classification with a molecular tool for menstrual cycle phase dating (beREADY®, CCHT). The program language R and Bioconductor packages were applied to analyse RNA-seq data in the setting of the result of accurate endometrial dating. To suggest robust candidate pathways, the identified differentially expressed genes (DEGs) associated with the adenomyosis group in the receptive phase were further integrated with 151, 173 and 42 extracted genes from published studies that were related to endometrial receptivity in healthy uterus, endometriosis and adenomyosis, respectively. Enrichment analyses were performed using Cytoscape ClueGO and CluePedia apps. Results Out of 20 endometrial samples, 2 were dated to the early receptive phase, 13 to the receptive phase and 5 to the late receptive phase. Comparison of the transcriptomics data from all 20 samples provided 909 DEGs (p Conclusions Accurate endometrial dating and RNA-seq analysis resulted in the identification of altered response to IFN signalling as the most promising candidate of impaired uterine receptivity in adenomyosis.

Inhaled corticosteroids (ICS) are the most common asthma controller medication. An important contribution of genetic factors in ICS response has been evidenced. Here, we aimed to identify novel genetic markers involved in ICS response in asthma. A genome-wide association study (GWAS) of the change in lung function after 6 weeks of ICS treatment was performed in 166 asthma patients from the SLOVENIA study. Patients with an improvement in lung function ≥8% were considered as ICS responders. Suggestively associated variants (p-value ≤ 5 × 10−6) were evaluated in an independent study (n = 175). Validation of the association with asthma exacerbations despite ICS use was attempted in European (n = 2681) and admixed (n = 1347) populations. Variants previously associated with ICS response were also assessed for replication. As a result, the SNP rs1166980 from the ROBO2 gene was suggestively associated with the change in lung function (OR for G allele: 7.01, 95% CI: 3.29–14.93, p = 4.61 × 10−7), although this was not validated in CAMP. ROBO2 showed gene-level evidence of replication with asthma exacerbations despite ICS use in Europeans (minimum p-value = 1.44 × 10−5), but not in admixed individuals. The association of PDE10A-T with ICS response described by a previous study was validated. This study suggests that ROBO2 could be a potential novel locus for ICS response in Europeans.

Objectives Combining of genomic data of different pathologies as a single phenotype has emerged as a useful strategy to identify genetic risk loci shared among immune-mediated diseases. Our study aimed to increase our knowledge of the genetic contribution to Kawasaki disease (KD) and IgA vasculitis (IgAV) by performing the first comprehensive large-scale analysis on the genetic overlap between them. Methods A total of 1190 vasculitis patients and 11 302 healthy controls were analysed. First, in the discovery phase, genome-wide data of 405 KD patients and 6252 controls and 215 IgAV patients and 1324 controls, all of European origin, were combined using an inverse variance meta-analysis. Second, the top associated polymorphisms were selected for replication in additional independent cohorts (570 cases and 3726 controls). Polymorphisms with P-values ≤5 × 10−8 in the global IgAV–KD meta-analysis were considered as shared genetic risk loci. Results A genetic variant, rs3743841, located in an intron of the NAGPA gene, reached genome-wide significance in the cross-disease meta-analysis (P = 8.06 × 10−10). Additionally, when IgAV was individually analysed, a strong association between rs3743841 and this vasculitis was also evident [P = 1.25 × 10−7; odds ratio = 1.47 (95% CI 1.27, 1.69)]. In silico functional annotation showed that this polymorphism acts as a regulatory variant modulating the expression levels of the NAGPA and SEC14L5 genes. Conclusion We identified a new risk locus with pleiotropic effects on the two childhood vasculitides analysed. This locus represents the strongest non-HLA signal described for IgAV to date.

Background In some patients securing the peripheral intravenous cannula (PIVC) with a standard adhesive dressing can be difficult because of sweat or other body fluids. The aim of our study was to evaluate the use of tissue adhesives alone as a means to secure PIVCs inserted in the emergency department. Methods We performed a prospective interventional pilot study from November 2019 to May 2020 in a medical emergency department of an urban tertiary hospital. Patients were randomized to two groups: tissue adhesives (TA) or adhesive dressing (AD) group. After randomization we followed them until day 4. Results There were no significant differences between TA and AD groups in the rate of unplanned removal of PIVCs in the first 72 h (57.1% vs. 45.8%, p = 0.29), the rate of unplanned removal of PIVCs in the ED (0% vs. 2.1%, p = 1.00), the rate of unplanned removal of PIVC in the first 24 h (42.8% vs. 35.4%, p = 0.52), as well as in the rate of phlebitis (7.1% vs. 14.6%, p = 0.34) and the rate of any blood-stream infection (0% vs. 0%, p = 1.00). Conclusion We did not observe any significant differences when PIVCs inserted in the emergency department were secured with tissue adhesives alone, compared to standard adhesive dressings. We observed a high rate of unplanned removal of PIVCs, necessitating further research to determine more reliable ways of securing PIVCs.

IntroductionBacterial community-acquired pneumonia (CAP) in children is caused mostly by Streptococcus pneumoniae. The resistance of pneumococci to penicillin is increasing. However, most guidelines still prefer treatment with narrow-spectrum antibiotics. Therefore, we compared the effect of intravenous treatment with penicillin and broad-spectrum β-lactam antibiotics in children with CAP. The objective of our study was to assess the eligibility of treatment of bacterial CAP with intravenous penicillin.Material and methodsWe performed a prospective study and included 136 children hospitalised because of bacterial CAP. Patients were treated intravenously with either penicillin G or broad-spectrum β-lactam antibiotic monotherapy. Lung ultrasound and blood tests were performed at admission and after 2 days of treatment. The time interval from the application of antibiotics to permanent defervescence was recorded.ResultsEighty-seven (64.0%) patients were treated with penicillin G, and 49 (36.0%) were treated with broad-spectrum β-lactam antibiotics. The median time to persistent defervescence was 5 h in the penicillin group and 8 h in the broad-spectrum group (p = 0.18). There were no significant differences between the treatment regimens regarding the effect on the consolidation sizes or C-reactive protein levels. However, the decrease in the white blood cell count was marginally greater in the penicillin treatment group (p = 0.05).ConclusionsWe have shown that penicillin is as effective as broad-spectrum antibiotics in the treatment of bacterial CAP in children. Although the resistance of pneumococci to antibiotics is increasing, clinicians should still use penicillin and other narrow-spectrum β-lactam antibiotics in the treatment of bacterial CAP in children.

Multiple sclerosis is an inflammatory disorder of the central nervous system. Inflammation may create high susceptibility to subclinical atherosclerosis. The purpose of this study was to compare subclinical atherosclerosis and the role of inflammatory cytokines between the group of patients with relapsing-remitting multiple sclerosis (RRMS) and healthy controls matched for age and sex. The study group consisted of 112 non-diabetic and non-hypertensive RRMS patients treated with disease modifying drugs (DMD) and the control group was composed of 51 healthy subjects. The common carotid artery (CCA) intima media thickness (IMT) was investigated. Serum levels of risk factors for atherosclerosis and inflammatory cytokines were also determined. The mean CCA IMT (0.572 ± 0.131 mm vs. 0.571 ± 0.114 mm) did not differ (p > 0.05) between patients and controls. The RRMS patients’ CCA IMT was significantly correlated with serum interleukin 6 (IL-6) (p = 0.027), high-sensitivity C-reactive protein (hs-CRP) (p = 0.027), cystatin C (p

Objectives Previous studies have found evidence for a genetic basis for pelvic organ prolapse (POP), but no genetic studies have differentiated between types of POP. This study investigated whether genetic variants in six previously suggested candidate loci for POP modify the risk of uterine prolapse (UP). Study design One hundred patients, aged 30–55 years, who had undergone surgery due to total UP and 105 healthy controls were included in this study. After extracting the genomic DNA from peripheral blood, six single nucleotide polymorphisms (SNPs) previously identified for POP were genotyped, and association analysis was performed for contributing risk factors. RNA expression was determined from sacrouterine ligaments of patients and controls using quantitative reverse transcription polymerase chain reaction. Results The dominant genotype model for the T allele for SNP rs6051098 at the chromosome 20p13 locus was significant, and this remained significant with the risk factor regression model (p=0.046; odds ratio 1.93, 95 % confidence interval 1.01–3.66). The isocitrate dehydrogenase 3 beta (IDH3B) gene was the only potential candidate gene in the 20p13 locus that was significantly upregulated in sacrouterine biopsies in women with UP compared with controls (p = 0.034). Conclusion To the best of the authors’ knowledge, this is the first study to show that genetic risk factors contribute to UP, and suggested rs6051098 as the best candidate risk factor associated with UP. According to expression data in sacrouterine tissue, this study suggests that the IDH3B gene plays a role in the pathogenesis of UP.

The success rate of percutaneous coronary artery intervention (PCI) of chronic total occlusion (CTO) lesions have increased in the recent years. However, improvement of function is only possible when significant myocardial viability is present. One of the most important factors of maintaining myocardial viability is the opening and development of collaterals. Our hypothesis was that with a higher degree of collaterals more viable myocardium is present. In 38 patients we compared the degree of collaterals, evaluated with a conventional coronary angiogram (CCA) and graded by the Rentrop classification to transmural extent of the scar obtained in a viability study with magnetic resonance (MRI). We found a statistically significant association of the degree of collaterals determined with Rentrop method and transmural extent of the scar as measured by CMR (p = 0.001; Tau = -0.144). Additionally, associations showed an increase in the ratio between viable vs. non-viable myocardium with the degree of collaterals. Our study suggests that it may be beneficial to routinely grade the collaterals at angiography in patients with CTO as an assessment of myocardial viability.

The aetiology of community-acquired pneumonia (CAP) is not easy to establish. As lung ultrasound (LUS) has already proved to be an excellent diagnostic tool for CAP, we analysed its usefulness for discriminating between the aetiologically different types of CAP in children. We included 147 children hospitalized because of CAP. LUS was performed in all patients at admission, and follow-up LUS was performed in most patients. LUS-detected consolidations in viral CAP were significantly smaller, with a median diameter of 15 mm, compared to 20 mm in atypical bacterial CAP (p = 0.05) and 30 mm in bacterial CAP (p

Response to anti-TNF therapy is crucial for life expectancy and life quality in patients with severe Crohn's disease. We investigated if a previously reported gene expression profile predictive for infliximab response could be also applied to adalimumab response in an independent cohort. Forty-seven Slovene Crohn's disease patients indicated for adalimumab therapy were enrolled in the study. Inflamed and non-inflamed colon biopsy samples were obtained during routine colonoscopy prior to adalimumab treatment. Response to adalimumab was measured with IBDQ. Gene expression in inflamed and non-inflamed colon biopsy samples was measured with RT-qPCR. Genotypes were extracted from previously available genotype data. Statistical analysis was performed with SPSS software. The R package e1071 was used to train bootstrap aggregated support vector machines (SVM). SVM prediction model analysis was used to analyze pooled, non-inflamed, and inflamed colon tissue datasets using IBDQ response after 4, 12, 20 and 30 weeks of adalimumab treatment. The bagging approach was used in an endeavor to obtain 100 % accuracy using 10 × 100 or 100 × 100 iterations. Average adalimumab response prediction accuracy is 75.5 % for pooled samples, 90.5 % for inflamed samples, and 100 % for non-inflamed samples. Moreover, models trained on selected SNPs from analyzed genes had an average accuracy of 92.8 %, confirming the involvement of genetic regions mapping the reported genes. Finally, using combined gene expression and SNP data we observed 100 % adalimumab response prediction accuracy for pooled, inflamed, and non-inflamed datasets. Our study supports the reported genetic anti-TNF response profile and extends it for adalimumab prediction. Eine ausreichende Antwort auf Anti-TNF-Behandlung mit Infliximab oder Adalimumab ist entscheidend für Lebenserwartung und Lebensqualität bei Patienten mit schwergradigem Morbus Crohn. Wir untersuchten, ob das zuvor beschriebene Genprofil zur Prognose des Therapieerfolgs mit Infliximab auch zur Prognose des Therapieerfolgs mit Adalimumab in einer unabhängigen Kohorte anwendbar ist. 47 slowenische für Adalimumab-Behandlung indizierte Morbus-Crohn-Patienten wurden in die Studie aufgenommen. Vor der Behandlung mit Adalimumab wurden während der Kolonoskopie entzündete und nicht entzündete Gewebeproben entnommen. Die Reaktion auf Adalimumab wurde mit IBDQ gemessen. Die Genexpression in Gewebeproben wurde mit RT-qPCR gemessen. Genotypen wurden aus zuvor verfügbaren Genotypdaten extrahiert. Die statistische Analyse wurde mit SPSS-Software durchgeführt. Das R-Paket e1071 wurde zum Trainieren von Support Vector Machines benutzt. Die SVM-Vorhersagemodelle wurden verwendet, um die gepoolten, nicht entzündeten und entzündeten Darmproben-Datensätze mit Verwendung von IBDQ-Antwort nach 4, 12, 20 und 30 Wochen Adalimumab-Behandlung zu analysieren. Die Bagging-Methode wurde verwendet, um eine 100-%-Genauigkeit mit Verwendung von 10 × 100 oder 100 × 100 Iterationen zu erreichen. Die Adalimumab-Antwortvorhersage von gepoolten Datensätzen erreichte im Durchschnitt eine Genauigkeit von 75,5 %, von entzündeten Datensätzen 90,5 % und von nicht entzündeten Datensätzen 100 %. Zusätzlich haben die Modelle, die an ausgewählten SNPs in den analysierten Genen trainiert wurden, eine durchschnittliche Genauigkeit von 92,8 % erreicht. Schließlich kombinierten wir Expressions- und Genotypdaten, um den Vorhersagewert weiter zu analysieren. Mithilfe kombinierter Daten konnten wir mit Darmgewebe-Datensätzen eine Adalimumab-Antwortvorhersage mit hundertprozentiger Sicherheit feststellen. Unsere Studie unterstützt das genetische Anti-TNF-Antwortprofil, das für den Adalimumab-Therapieerfolg gleichermaßen relevant ist.

Introduction: Inhaled corticosteroids (ICS) are the most effective medication for control of asthma symptoms. However, a proportion of patients do not respond to this medication and suffer exacerbations, due to a combination of environmental and genetic factors. We performed a meta- genome-wide association study (GWAS) to identify genes associated with ICS response in European children. Methods: A GWAS meta-analysis of asthma exacerbations was performed across eight European studies from the Pharmacogenomics in Childhood Asthma (PiCA) consortium including 2,704 asthmatic children and young adults treated with ICS. Imputation of genetic variants was carried out using the Haplotype Reference Consortium as reference panel and association testing was performed by means of logistic regression models. A total of 8.1 million genetic variants with minor allele frequency ≥1% were meta-analyzed. Results: Nineteen polymorphisms were suggestively associated with asthma exacerbations despite use of ICS (p≤5x10-6). These variants were located at 10 segregating sites (minimum p=6.7x10-7). Novel associations were revealed in biologically plausible genes involved in drug metabolism (AOX1) and previously associated with lung function measurements (WNT5A and CNTNAP5). Moreover, variants located at CNTNAP5 were also nominally associated with a change in FEV1 after ICS treatment in SLOVENIA (minimum p=0.025). Conclusions: We revealed genes suggestively associated with asthma exacerbations in children and young adults despite the use of ICS. Replication will be performed in further independent studies. Study supported by Instituto de Salud Carlos III (AC15/00015) and ERACoSysMed 1st Joint Transnational Call (SysPharmPedia).

Background\ud Inhaled corticosteroids (ICS) are the most widely prescribed and effective medication to control asthma symptoms and exacerbations. However, many children still have asthma exacerbations despite treatment, particularly in admixed populations, such as Puerto Ricans and African Americans. A few genome‐wide association studies (GWAS) have been performed in European and Asian populations, and they have demonstrated the importance of the genetic component in ICS response.\ud \ud Objective\ud We aimed to identify genetic variants associated with asthma exacerbations in admixed children treated with ICS and to validate previous GWAS findings.\ud \ud Methods\ud A meta‐analysis of two GWAS of asthma exacerbations was performed in 1347 admixed children treated with ICS (Hispanics/Latinos and African Americans), analysing 8.7 million genetic variants. Those with P ≤ 5 × 10−6 were followed up for replication in 1697 asthmatic patients from six European studies. Associations of ICS response described in published GWAS were followed up for replication in the admixed populations.\ud \ud Results\ud A total of 15 independent variants were suggestively associated with asthma exacerbations in admixed populations (P ≤ 5 × 10−6). One of them, located in the intergenic region of APOBEC3B and APOBEC3C, showed evidence of replication in Europeans (rs5995653, P = 7.52 × 10−3) and was also associated with change in lung function after treatment with ICS (P = 4.91 × 10−3). Additionally, the reported association of the L3MBTL4‐ARHGAP28 genomic region was confirmed in admixed populations, although a different variant was identified.\ud \ud Conclusions and clinical relevance\ud This study revealed the novel association of APOBEC3B and APOBEC3C with asthma exacerbations in children treated with ICS and replicated previously identified genomic regions. This contributes to the current knowledge about the multiple genetic markers determining responsiveness to ICS which could lead in the future the clinical identification of those asthma patients who are not able to respond to such treatment.

Summary Background & aims Celiac disease (CD) is an immune-mediated systemic disease, caused by ingestion of gluten in genetically predisposed individuals. Gut microbiota dysbiosis might play a significant role in pathogenesis of chronic enteropathies and its modulation can be used as an intervention strategy in CD as well. In this study, we aimed to identify correlations between fecal microbiota, serum tumor necrosis factor alpha (TNF-α) and fecal short-chain fatty acids (SCFAs) in healthy children and children with CD after administration of probiotic Bifidobacterium breve BR03 and B632. Methods A double-blind placebo-controlled study enrolled 40 children with CD (CD) and 16 healthy children (HC). CD children were randomly allocated into two groups, of which 20 belonged to the placebo (PL) group and 20 to the Probiotic (PR) group. The PR group received a probiotic formulation containing a mixture of 2 strains, B. breve BR03 (DSM 16604) and B. breve B632 (DSM 24706) in 1:1 ratio for 3 months. Subsequently, for statistical analysis, blood and fecal samples from CD children (on enrolment - T0 and after 3 months, at the end of intervention with probiotic/placebo - T1) and HC children were used. The HC group was sampled only once (T0). Results Verrucomicrobia, Parcubacteria and some yet unknown phyla of Bacteria and Archaea may be involved in the disease, indicated by a strong correlation to TNF-α. Likewise, Proteobacteria strongly correlated with fecal SCFAs concentration. The effect of probiotic administration has disclosed a negative correlation between Verrucomicrobia, some unknown phyla of Bacteria, Synergistetes, Euryarchaeota and some SCFAs, turning them into an important target in microbiome restoration process. Synergistetes and Euryarchaeota may have a role in the anti-inflammatory process in healthy human gut. Conclusions Our results highlight new phyla, which may have an important relation to disease-related parameters, CD itself and health.

The early establishment of a complete microbiome has been shown to play an integral part in the development and maintenance of an intact intestine and its immune system, although much remains unknown about the specific mechanisms of immune modulation in newborns. In our study we show in a co-culture model of the undeveloped small intestine that members of Lactobacillus spp. influence STAT1 and NF-kB p65 nuclear translocation in both intestinal epithelial cells as well as underlying macrophages. Moreover, by using imaging flow cytometry we were able to monitor each individual cell and create a framework of the percentage of cells in which translocation occurred in challenged versus control cell populations. We also observed a significant difference in baseline translocation in intestinal cells when cultured alone versus those in a co-culture model, underpinning the importance of 3D models over monolayer set-ups in epithelial in vitro research. In conclusion, our work offers new insights into the potential routes by which the commensal microbiome primes the early immune system to fight pathogens, and shows how strain-specific these mechanisms really are.

World climate, environmental and other global changes would have wide-ranging and, mostly, adverse consequences for human health. These changes are a significant and emerging threat to public health, including the growing season, variability of crop yields and water demands. Above mentioned factors have been influenced in areas of food security and food safety and moreover the food industry struggle for profits. These factors control the conditions and affect the use of chemicals in the agriculture and food industry. In this chapter, the risk assessment of chemicals in food as a part of risk analysis process have been addressed. We would like to point out the importance of this process in the area of public health and consumer protection. Through food and nutrition products and most often misleading advertisements, people are exposed to different hazards that could have severe harmful consequences. Among the consumer groups, children are mostly vulnerable because of their adult dependencies, age and body weight. In this chapter, an example of risk assessment of exposure to food additives among preschool children in Slovenia is evaluated.