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The incidence of the autoimmune disease type 1 diabetes is increasing, likely caused by environmental factors. A gluten-free diet has previously been shown to ameliorate autoimmune diabetes in non-obese diabetic (NOD) mice and humans. Although the exact mechanisms are not understood, interventions influencing the intestinal microbiota early in life affect the risk of type 1 diabetes. Here, we characterize how NOD mice that are fed a gluten-free (GF) diet differ from NOD mice that are fed a gluten-containing standard (STD) diet in terms of their microbiota composition by 16S rRNA gene amplicon sequencing and pancreatic immune environment by real-time quantitative PCR at the prediabetic stage at 6 and 13 weeks of age. Gut microbiota analysis revealed highly distinct microbiota compositions in both the cecum and the colon of GF-fed mice compared with STD-fed mice. The microbiotas of the GF-fed mice were characterized by an increased Firmicutes/Bacteroidetes ratio, an increased abundance of short chain fatty acid (particularly butyrate)-producing bacteria, and a reduced abundance of Lactobacilli compared with STD mice. We found that the insulitis score in the GF mice was significantly reduced compared with the STD mice and that the markers for regulatory T cells and T helper 2 cells were upregulated in the pancreas of the GF mice. In conclusion, a GF diet during pre- and early post-natal life induces shifts in the cecal and colonic microbiota compatible with a less inflammatory environment, providing a likely mechanism for the protective effect of a GF diet in humans.

Introduction: As part of the study CODIBINE, Correlations and Diagnoses for Biomarkers in New-borns, the main objective of the study was to explore neonatal inflammation, stress, neurodevelopment, and growth factors after in-labor and pre-labor cesarean section compared to vaginal delivery. Increasing evidence has shown that birth delivery mode has an impact on imminent and long-term child health. However, the effect of the timing of cesarean section is insufficiently elucidated. The main objective of the study was to explore the effect of different delivery modes, vaginal delivery compared to cesarean section with or without initiation of labor, on the infants. Methods: We designed a retrospective cohort study, including dried blood spot samples from mature (gestational age ≥ 37) newborns delivered in the years 2009-2011. The newborns were divided into three groups after delivery mode: (1) pre-labor cesarean section (n = 714), i.e., cesarean delivery without initiation of labor, (2) in-labor cesarean section (n = 655), i.e., cesarean section after initiation of labor, and (3) vaginal delivery (n = 5,897). We measured infant levels of inflammatory (IL-18, MCP-1, CRP, sTNF RI), stress (HSP-70), growth (EGF, VEGF-A), and neurotrophic factors (BDNF, NT-3, S100B) 2–4 days after birth. Results: The neonatal levels of inflammatory and stress markers were significantly lower, while the levels of growth factors were higher after pre-labor cesarean section compared to vaginal delivery. The biomarker levels were similar after in-labor cesarean section and vaginal delivery. Removing cases with pre-labor rupture of membranes and artificial rupture of membranes in the calculations did not change the results. The levels of neurotrophic factors were unaffected by delivery form. Males had generally higher levels of inflammation and lower levels of growth and neurotrophic factors. Overall, the levels of inflammatory markers increased, and the growth factors decreased with increas

Background/Objectives: Pre-treatment gut microbial Prevotella-to-Bacteroides (P/B) ratio and markers of glucose metabolism (i.e., fasting glucose and insulin) have been suggested as biomarkers for optimal weight management. However, both biomarkers need further validation, and the interactions between them for optimal weight management are largely unknown. To investigate differences in weight loss maintenance between subjects with low and high P/B ratio and the potential interactions with markers of glucose metabolism and dietary fiber intake.Subjects/Methods: Following an 8-week weight loss period using meal replacement products, subjects losing ≥ 8% of their initial body weight were randomized to one of three protein supplements or maltodextrin for a 24-week weight maintenance period. Habitual diet was consumed along with the supplements expected to constitute 10-15% of total energy. For this analysis we stratified the participants into low and high strata based on median values of pre-intervention P/B ratio, pre-weight loss Homeostatic model assessment of insulin resistance (HOMA-IR) (<2.33 or > 2.33), and dietary fiber intake during the intervention (< 28.5 or > 28.5 g/10 MJ).Results: Regardless of weight maintenance regimen, subjects with high P/B ratio (n = 63) regained 1.5 (95% CI 0.4, 2.7) kg body weight (P = 0.007) more than subjects with low P/B ratio (n = 63). The regain among subjects with high P/B ratio was particular evident if HOMA-IR was high and dietary fiber intake was low. Consequently, in the high P/B strata, subjects with high HOMA-IR and low fiber intake (n = 17) regained 5.3 (95% CI 3.3, 7.3) kg (P < 0.001) more body weight compared with participants with low HOMA-IR and high fiber intake (n = 16).Conclusions: Subjects with high P/B ratio were more susceptible to rega

Scope: The impact of dietary protein types on the gut microbiome is scarcely studied. The aim of the present study is therefore to examine the effects of lean-seafood and non-seafood proteins on the gut microbiome composition and activity and elucidate potential associations to cardiovascular disease (CVD) risk factors. Methods: A crossover intervention study in which 20 healthy subjects consumed two diets that varied in protein source was conducted. 1H NMR spectroscopy and 16S rDNA sequencing analyses were applied to characterize fecal metabolites and gut microbiota composition, respectively. Results: A twofold increase in fecal trimethylamine excretion was observed after the lean-seafood diet period. Circulating TAG and the total to high-density lipoprotein (HDL) cholesterol ratio as well as circulating TMAO levels were each associated with specific gut bacteria. Following the non-seafood diet period, a decreased relative abundance of Clostridium cluster IV and a tendency toward an increased Firmicutes/Bacteroidetes ratio were found. Conclusions: Lean-seafood and non-seafood diets differentially modulate the gut microbiome composition and activity. Furthermore, the gut microbiota composition seems to affect circulating TMAO levels and CVD risk factors.

Background/objectives: Individuals with high pre-treatment bacterial Prevotella-to-Bacteroides (P/B) ratio have been reported to lose more body weight on diets high in fiber than subjects with a low P/B ratio. Therefore, the aim of the present study was to examine potential differences in dietary weight loss responses between participants with low and high P/B.Subjects/methods: Eighty overweight participants were randomized (52 completed) to a 500 kcal/d energy deficit diet with a macronutrient composition of 30 energy percentage (E%) fat, 52 E% carbohydrate and 18 E% protein either high (≈1500 mg calcium/day) or low ( ≤ 600 mg calcium/day) in dairy products for 24 weeks. Body weight, body fat, and dietary intake (by 7-day dietary records) were determined. Individuals were dichotomized according to their pre-treatment P/B ratio derived from 16S rRNA gene sequencing of collected fecal samples to test the potential modification of dietary effects using linear mixed models.Results: Independent of the randomized diets, individuals with high P/B lost 3.8 kg (95%CI, 1.8,5.8; P < 0.001) more body weight and 3.8 kg (95% CI, 1.1, 6.5; P = 0.005) more body fat compared to individuals with low P/B. After adjustment for multiple covariates, individuals with high P/B ratio lost 8.3 kg (95% CI, 5.8;10.9, P < 0.001) more body weight when consuming above compared to below 30 g fiber/10MJ whereas this weight loss was 3.2 kg (95% CI, 0.8;5.5, P = 0.008) among individuals with low P/B ratio [Mean difference: 5.1 kg (95% CI, 1.7;8.6, P = 0.003)]. Partial correlation coefficients between fiber intake and weight change was 0.90 (P < 0.001) among individuals with high P/B ratio and 0.25 (P = 0.29) among individuals with low P/B ratio.Conclusions: Individuals with high P/B lost more body weight and body fat compared to individuals with low P/B, confirmin

During weight loss, dairy calcium is proposed to accelerate weight and fat mass loss through increased fecal fat excretion. The primary objective was to investigate if a high-dairy energy-restricted diet is superior to low-dairy in terms of changes in body weight, body composition and fecal fat excretion over 24 weeks. Secondary objectives included fecal energy and calcium excretion, resting energy expenditure, blood pressure, lipid metabolism and gut microbiota. In a randomized, parallel-arm intervention study 11 men and 69 women (BMI 30.60.3 kg/m2, age 441 years) were allocated to a 500 kcal (2100 kJ) deficit diet either high (HD: 1500 mg calcium/d) or low (LD: 600 mg calcium/d) in dairy products for 24 weeks. Habitual calcium intake was ~1000 mg/d. Body weight loss (HD: -6.6±1.3 kg, LD: -7.9±1.5 kg, P=0.73), fat mass loss (HD: -7.8±1.3 %, LD: -8.5±1.1 %, P=0.76), changes in fecal fat excretion (HD: -0.57±0.76 g, LD: 0.46±0.70 g, P=0.12) and microbiota composition were similar for the groups over 24 weeks. However, total fat mass loss was positively associated with relative abundance of Papillibacter (P=0.017) independent of diet group. Consumption of a high dairy diet did not increase fecal fat or accelerate weight and fat mass loss beyond energy restriction over 24 weeks in overweight and obese adults with a habitual calcium intake of ~1000 mg/d. However, this study indicate that Papillibacter is involved in body compositional changes.

Cheese and butter have been shown to affect blood lipids differently. This parallel-arm, randomised, controlled study in 36 crossbred growing sows compared the effect of diets with either regular-fat cheese (REG), reduced-fat cheese + butter (RED) or butter (BUT) on blood lipids, faecal fat and energy excretion and gut microbiome in pigs. A 14-d run-in period was followed by 14-d interventions with macronutrient-matched diets. Fasting total cholesterol and HDL-cholesterol after 14 days were higher in REG compared with BUT, but only tended to be higher in RED. Compared with BUT, REG and RED had higher faecal fat excretion. Faecal energy excretion was only higher in REG, and this correlated with a lower microbiome Firmicutes-to-Bacteroidetes ratio. In conclusion, dairy fat consumed as cheese or butter caused different metabolic effects. Differences between reduced-fat cheese+butter and butter were less pronounced than differences between regular-fat cheese and butter, suggesting an impact of the dairy-matrix.

Disseminated melanoma is a serious disease, causing the death of thousands of people each year. Advanced stages of the disease are not sensitive to conventional therapies, however new immunotherapy treatments are emerging. The novel treatment of adaptive cell therapy (ACT) has shown partial responses in about 50% of patients with metastatic melanoma. A key reason for lack of efficacy in some patients is immunosuppression by the tumor microenvironment. Targeting of immunosuppressive molecules could therefore be very beneficial in the treatment of advanced melanoma and, when combined with ACT, could lead to a larger fraction of cured patients. MMP23 is a metalloproteinase expressed by a subset of melanomas. It has been shown that tumors expressing high levels of MMP23 have fewer tumor-infiltrating lymphocytes (TIL) and that there is correlation between high levels of MMP23 and an overall worse clinical outcome. MMP23 is a unique metalloproteinase, and has four protein domains, one of which is a ShK toxin-like domain that has been shown to block the Kv1.3 potassium channel. T cells express this channel, especially effector memory T cells, which upregulate it when activated. We hypothesize that melanoma cells can suppress effector memory T cells by upregulating MMP23, which can block Kv1.3 potassium channels on the T cells, and thereby inhibit the anti-tumor response. In this project we have established the use of CRISPR/Cas9 for characterization and targeting of novel inhibitory mechanisms in the tumor microenvironment. Using the CRISPR/Cas9 technique, we have successfully created and validated two guide RNAs (gRNAs) for MMP-23. These gRNAs target exon 5 and exon 7 of the MMP23B gene, and by co-transfecting with both, we have effectively knocked out exon 6, which encodes the ShK toxin-like domain. We have thereby created MMP23 knockout tumor cell lines, hopefully incapable of bl