The chemokine CXCL13 is used as complement to serology in the diagnostics of Lyme neuroborreliosis (LNB). We evaluated and compared the semi-quantitative, cassette-based ReaScan CXCL13 assay with the quantitative recomBead CXCL13 assay using a collection of 209 cerebrospinal fluid samples. The categorical agreement between results interpreted as negative, grey zone, and positive by the two methods was 87%. The diagnostic sensitivity was higher using the recomBead assay, whereas specificity was higher using ReaScan. Few manual steps, and a short turn-around time with no batching of samples makes the ReaScan CXCL13 assay an attractive complement to serology in the diagnostics of LNB.

Vedolizumab is efficacious in the treatment of Crohn’s disease (CD) and ulcerative colitis (UC). However, a significant proportion of patients present with a non-response. To investigate whether differences in the clinical response to vedolizumab is reflected in changes in gene expression levels in whole blood, samples were collected at baseline before treatment, and at follow-up after 10–12 weeks. Whole genome transcriptional profiles were established by RNA sequencing. Before treatment, no differentially expressed genes were noted between responders (n = 9, UC 4, CD 5) and non-responders (n = 11, UC 3, CD 8). At follow-up, compared with baseline, responders displayed 201 differentially expressed genes, and 51 upregulated (e.g., translation initiation, mitochondrial translation, and peroxisomal membrane protein import) and 221 downregulated (e.g., Toll-like receptor activating cascades, and phagocytosis related) pathways. Twenty-two of the upregulated pathways in responders were instead downregulated in non-responders. The results correspond with a dampening of inflammatory activity in responders. Although considered a gut-specific drug, our study shows a considerable gene regulation in the blood of patients responding to vedolizumab. It also suggests that whole blood is not optimal for identifying predictive pre-treatment biomarkers based on individual genes. However, treatment outcomes may depend on several interacting genes, and our results indicate a possible potential of pathway analysis in predicting response to treatment, which merits further investigation.

Combination treatment with low-dose thiopurine and allopurinol (AP) has successfully been used in patients with inflammatory bowel disease with a so called skewed thiopurine metabolite profile. In red blood cells in vivo, it reduces the concentration of methylated metabolites and increases the concentration of the phosphorylated ones, which is associated with improved therapeutic efficacy. This study aimed to investigate the largely unknown mechanism of AP on thiopurine metabolism in cells with an active thiopurine metabolic pathway using HepG2 and HEK293 cells. Cells were treated with 6-mercaptopurine (6MP) and AP or its metabolite oxypurinol. The expression of genes known to be associated with thiopurine metabolism, and the concentration of thiopurine metabolites were analyzed. Gene expression levels were only affected by AP in the presence of 6MP. The addition of AP to 6MP affected the expression of in total 19 genes in the two cell lines. In both cell lines the expression of the transporter SLC29A2 was reduced by the combined treatment. Six regulated genes in HepG2 cells and 8 regulated genes in HEK293 cells were connected to networks with 18 and 35 genes, respectively, present at known susceptibility loci for inflammatory bowel disease, when analyzed using a protein-protein interaction database. The genes identified as regulated as well as the disease associated interacting genes represent new candidates for further investigation in the context of combination therapy with thiopurines and AP. However, no differences in absolute metabolite concentrations were observed between 6MP+AP or 6MP+oxypurinol vs. 6MP alone in either of the two cell lines. In conclusion; the effect of AP on gene expression levels requires the presence of 6MP, at least in vitro. Previously described AP-effects on metabolite concentrations observed in red blood cells in vivo could not be reproduced in our cell lines in vitro. AP’s effects in relation to thiopurine metabolism are complex. The network-identified susceptibility genes represented biological processes mainly associated with purine nucleotide biosynthetic processes, lymphocyte proliferation, NF-KB activation, JAK-STAT signaling, and apoptotic signaling at oxidative stress.

Background:Identification of high-risk stage II colorectal cancer (CRC) patients, potential candidates for adjuvant chemotherapy, is challenging. Current clinical guidelines rely mainly on histopathological markers with relatively weak prognostic value. This motivates further search for prognostic markers. Methods:This explorative study aimed to identify potential candidate gene mutations to facilitate differentiation between subgroups of patients with CRC stage II. Panel-based massive parallel sequencing was used to genetically characterize tumor tissues from 85 patients radically operated for CRC stage II, of which 12 developed recurrent cancer during follow-up. Genetic data was compared between patients with or without cancer recurrence, between tumors located in colon and in rectum, and for association with tumor differentiation grade. Results:Genetic variation in ATM, C11ORF65 was associated with recurrence-free survival. Previous reports regarding the association between BRAF mutation and a higher age at diagnosis, and tumor location in colon were confirmed. APC, BRAF, or KRAS mutation was associated with tumor differentiation grade. Multiple correspondence analyses revealed no obvious clustering of patients with the studied clinical characteristics, indicating that the genetic signatures observed here were unique for each individual. Conclusions:Taken together, we have demonstrated the utility of panel-based massive parallel sequencing to explore the pathogenesis of CRC stage II. We have identified promising candidate gene mutations associated with cancer recurrence, tumor location, and differentiation grade in patients with CRC stage II, which merit further investigation.

Lyme neuroborreliosis (LNB) is the most common form of disseminated Lyme borreliosis in Europe and North America. There are limitations in existing LNB diagnostics and a lack of reliable objective markers for disease-course. Here, extensive protein profiling with two panels of 184 proteins, was done in the search for new clinically useful diagnostic and prognostic candidate biomarkers. Cerebrospinal fluid (CSF) was collected from patients with definite LNB (n = 13) at the time of diagnosis before initiating antibiotic treatment, and at a follow-up one month later. When symptoms were evaluated at a six-month follow-up, six patients had recovered with no persistent symptoms (NPS), and seven experienced delayed recovery with persistent post-treatment symptoms (PS). Orthopedic patients (n = 60) served as controls. With the panels used, no protein biomarkers able to differentiate between PS and NPS were identified. However, from a diagnostic perspective, we identified multiple proteins that were differentially expressed between LNB and controls. The majority of them were downregulated following antibiotic treatment, at the one-month follow-up. IL10, TNF, and CCL8 were considered examples of potentially useful candidate biomarkers in both the early diagnostics and in monitoring of treatment response. These markers merit further investigation to understand their utility in relation to other neurological manifestations.

A rapid and reliable influenza method is important to meet diagnostic needs. Here, the performance of the cartridge-based Xpert Flu assay was evaluated in comparison with an in-house real-time PCR method. Analysis was then decentralized from the clinical microbiology laboratory located at County Hospital Ryhov, Jönköping, Sweden, to the clinical chemistry laboratories present at the three hospitals in Region Jönköping County, allowing a 24/7 service. In total, 189 clinical samples were included in 2014-2015. The overall sensitivity and specificity of Xpert Flu were both 99%. With Xpert Flu and the 24/7 service the median turn-around time from sampling to available test result decreased from 22 h in 2013 to 1.9 h in 2015. In summary, the analytical performance of Xpert Flu is excellent. The large degree of automation of GeneXpert and Xpert Flu with assay on demand has allowed a 24/7 service at the clinical chemistry laboratories in the county, and a significantly reduced turn-around time, which is valuable for both patients and the healthcare providers.