Nonobese diabetic (NOD) mice are a widely used animal model to study mechanisms leading to autoimmune diabetes. A gluten-free diet reduces and delays the incidence of diabetes in NOD mice, but the underlying mechanisms remain largely unknown. In this study, we performed single-cell transcriptomic and flow cytometry analysis of T cells and innate lymphocytes in the spleen and pancreatic lymph nodes of NOD mice fed a gluten-free or standard diet. We observed that the gluten-free diet did not induce a substantial alteration in the abundance or phenotype of any lymphocyte subset that would directly explain its protective effect against diabetes. However, the gluten-free diet induced subtle changes in the differentiation of subsets with previously proposed protective roles in diabetes development, such as Tregs, activated γδT cells, and NKT cells. Globally, the gluten-free diet paradoxically promoted activation and effector differentiation across multiple subpopulations and induced genes regulated by IL-2, IL-7, and IL-15. In contrast, the standard diet induced type I interferon-responsive genes. Overall, the gluten-free diet might prevent diabetes in NOD mice by inducing small-scale changes in multiple cell types rather than acting on a specific lymphocyte subset.

• Keywords
• NOD mice, T regulatory cells, gluten‐free diet, single‐cell transcriptomics, type I diabetes,
• MeSH
• Lymphocyte Activation immunology   MeSH
• Diet, Gluten-Free * MeSH
• Cell Differentiation MeSH
• Diabetes Mellitus, Type 1 * immunology   MeSH
• Mice, Inbred NOD MeSH
• Mice MeSH
• T-Lymphocyte Subsets * immunology   MeSH
• Transcriptome MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

• Keywords
• NOD mouse, beta-cell stress, enterovirus, environmental factors, gluten-free diet, gut-pancreas axis, intestinal permeability, type 1 diabetes,
• MeSH
• Abdomen MeSH
• Diabetes Mellitus, Type 1 * MeSH
• Humans MeSH
• Pancreas MeSH
• Check Tag
• Humans MeSH
• Publication type
• Editorial MeSH

AIMS/HYPOTHESIS: The proportion of children with type 1 diabetes (T1D) who have experience with low-carbohydrate diet (LCD) is unknown. Our goal was to map the frequency of LCD among children with T1D and to describe their clinical and laboratory data. METHODS: Caregivers of 1040 children with T1D from three centers were addressed with a structured questionnaire regarding the children's carbohydrate intake and experience with LCD (daily energy intake from carbohydrates below 26% of age-recommended values). The subjects currently on LCD were compared to a group of non-LCD respondents matched to age, T1D duration, sex, type and center of treatment. RESULTS: A total of 624/1040 (60%) of the subjects completed the survey. A total of 242/624 (39%) subjects reported experience with voluntary carbohydrate restriction with 36/624 (5.8%) subjects currently following the LCD. The LCD group had similar HbA1c (45 vs. 49.5, p = 0.11), lower average glycemia (7.0 vs. 7.9, p = 0.02), higher time in range (74 vs. 67%, p = 0.02), lower time in hyperglycemia >10 mmol/L (17 vs. 20%, p = 0.04), tendency to more time in hypoglycemia <3.9 mmol/L(8 vs. 5%, p = 0.05) and lower systolic blood pressure percentile (43 vs. 74, p = 0.03). The groups did not differ in their lipid profile nor in current body height, weight or BMI. The LCD was mostly initiated by the parents or the subjects themselves and only 39% of the families consulted their decision with the diabetologist. CONCLUSIONS/INTERPRETATION: Low carbohydrate diet is not scarce in children with T1D and is associated with modestly better disease control. At the same time, caution should be applied as it showed a tendency toward more frequent hypoglycemia.

• Keywords
• low-carbohydrate diet, time in range, type 1 diabetes,
• MeSH
• Diabetes Mellitus, Type 1 diet therapy   metabolism   MeSH
• Diet, Carbohydrate-Restricted * adverse effects   statistics & numerical data   MeSH
• Child MeSH
• Glycated Hemoglobin analysis   MeSH
• Body Mass Index MeSH
• Blood Glucose analysis   MeSH
• Humans MeSH
• Lipids blood   MeSH
• Surveys and Questionnaires MeSH
• Body Weight MeSH
• Body Height MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Multicenter Study MeSH
• Names of Substances
• Glycated Hemoglobin A MeSH
• Blood Glucose MeSH
• Lipids MeSH

BACKGROUND: Blastocystis is a human gut symbiont of yet undefined clinical significance. In a set of faecal samples collected from asymptomatic children of six distant populations, we first assessed the community profiles of protist 18S rDNA and then characterized Blastocystis subtypes and tested Blastocystis association with the faecal bacteriome community. METHODS: Stool samples were collected from 244 children and young persons (mean age 11.3 years, interquartile range 8.1-13.7) of six countries (Azerbaijan 51 subjects, Czechia 52, Jordan 40, Nigeria 27, Sudan 59 and Tanzania 15). The subjects showed no symptoms of infection. Amplicon profiling of the 18S rDNA was used for verification that Blastocystis was the most frequent protist, whereas specific real-time PCR showed its prevalence and quantity, and massive parallel amplicon sequencing defined the Blastocystis subtypes. The relation between Blastocystis and the stool bacteriome community was characterized using 16S rDNA profiling. RESULTS: Blastocystis was detected by specific PCR in 36% (88/244) stool samples and was the most often observed faecal protist. Children from Czechia and Jordan had significantly lower prevalence than children from the remaining countries. The most frequent subtype was ST3 (49%, 40/81 sequenced samples), followed by ST1 (36%) and ST2 (25%). Co-infection with two different subtypes was noted in 12% samples. The faecal bacteriome had higher richness in Blastocystis-positive samples, and Blastocystis was associated with significantly different community composition regardless of the country (p < 0.001 in constrained redundancy analysis). Several taxa differed with Blastocystis positivity or quantity: two genera of Ruminococcaceae were more abundant, while Bifidobacterium, Veillonella, Lactobacillus and several other genera were undrerrepresented. CONCLUSIONS: Asymptomatic children frequently carry Blastocystis, and co-infection with multiple distinct subtypes is not exceptional. Prevalence and quantity of the organism clearly differ among populations. Blastocystis is linked to both faecal bacteriome diversity and its composition.

• Keywords
• Africa, Asia, Bacteriome, Blastocystis, Type 1 diabetes,
• MeSH
• Asymptomatic Infections epidemiology   MeSH
• Blastocystis classification   genetics   isolation & purification   MeSH
• Blastocystis Infections epidemiology   parasitology   MeSH
• Child MeSH
• Feces parasitology   MeSH
• Genetic Variation MeSH
• Humans MeSH
• Adolescent MeSH
• Prevalence MeSH
• DNA, Protozoan genetics   MeSH
• DNA, Ribosomal genetics   MeSH
• Gastrointestinal Microbiome genetics   MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Azerbaijan epidemiology   MeSH
• Czechoslovakia epidemiology   MeSH
• Jordan epidemiology   MeSH
• Nigeria epidemiology   MeSH
• Sudan epidemiology   MeSH
• Tanzania epidemiology   MeSH
• Names of Substances
• DNA, Protozoan MeSH
• DNA, Ribosomal MeSH