Interaction between GPR120 p.R270H loss-of-function variant and dietary fat intake on incident type 2 diabetes risk in the D.E.S.I.R. study.

BACKGROUND AND AIMS: GPR120 (encoded by FFAR4) is a lipid sensor that plays an important role in the control of energy balance. GPR120 is activated by long chain fatty acids (FAs) including omega-3 FAs. In humans, the loss of function p.R270H variant of the gene FFAR4 has been associated with a lower protein activity, an increased risk of obesity and higher fasting plasma glucose levels. The aim of this study was to investigate whether p.R270H interacts with dietary fat intake to modulate the risk of type 2 diabetes (T2D, 198 incident; 368 prevalent cases) and overweight (787 incident and 2891 prevalent cases) in the prospective D.E.S.I.R. study (n = 5,212, 9 years follow-up). METHODS AND RESULTS: The association of p.R270H with dietary fat and total calories was assessed by linear mixed models. The interaction between p.R270H and dietary fat on T2D and overweight was assessed by logistic regression analysis. The p.R270H variant had a minor allele frequency of 1.45% and was not significantly associated with total calories intake, fat intake or the total calories derived from fat (%). However, there was a significant interaction between p.R270H and dietary fat modulating the incidence of T2D (Pinteraction = 0.02) where the H-carriers had a higher risk of T2D than RR homozygotes in the low fat intake category only. The interaction between p.R270H and fat intake modulating the incidence and prevalence of overweight was not significant. CONCLUSION: The p.R270H variant of GPR120 modulates the risk of T2D in interaction with dietary fat intake in the D.E.S.I.R.

A nonsense loss-of-function mutation in PCSK1 contributes to dominantly inherited human obesity.

BACKGROUND: A significant proportion of severe familial forms of obesity remain genetically elusive. Taking advantage of our unique cohort of multigenerational obese families, we aimed to assess the contribution of rare mutations in 29 common obesity-associated genes to familial obesity, and to evaluate in these families the putative presence of nine known monogenic forms of obesity. METHODS: Through next-generation sequencing, we sequenced the coding regions of 34 genes involved in polygenic and/or monogenic forms of obesity in 201 participants (75 normal weight individuals, 54 overweight individuals and 72 individuals with obesity class I, II or III) from 13 French families. In vitro functional analyses were performed to investigate the mutation PCSK1-p.Arg80* which was identified in a family. RESULTS: A novel heterozygous nonsense variant in PCSK1 (p.Arg80*), encoding a propeptide truncated to less than two exons (out of 14), was found to co-segregate with obesity in a three-generation family. We demonstrated that this mutation inhibits PCSK1 enzyme activity and that this inhibition most likely does not involve a strong physical interaction. Furthermore, both mutations PCSK1-p.Asn180Ser and POMC-p.Phe144Leu, which had previously been reported to be associated with severe obesity, were also identified in this study, but did not co-segregate with obesity. Finally, we did not identify any rare mutations co-segregating with obesity in common obesity susceptibility genes, except for CADM2 and QPCTL, where we found two novel variants (p.Arg81His and p.Leu98Pro, respectively) in three obese individuals. CONCLUSIONS: We showed for the first time that a nonsense mutation in PCSK1 was likely to cause dominantly inherited human obesity, due to the inhibiting properties of the propeptide fragment encoded by the null allele. Furthermore, the present family sequencing design challenged the contribution of previously reported mutations to monogenic or at least severe obesity.

Multiple functional polymorphisms in the G6PC2 gene contribute to the association with higher fasting plasma glucose levels.

AIMS/HYPOTHESIS: We previously identified the G6PC2 locus as a strong determinant of fasting plasma glucose (FPG) and showed that a common G6PC2 intronic single nucleotide polymorphism (SNP) (rs560887) and two common G6PC2 promoter SNPs (rs573225 and rs13431652) are highly associated with FPG. However, these promoter SNPs have complex effects on G6PC2 fusion gene expression, and our data suggested that only rs13431652 is a potentially causative SNP. Here we examine the effect of rs560887 on G6PC2 pre-mRNA splicing and the contribution of an additional common G6PC2 promoter SNP, rs2232316, to the association signal. METHODS: Minigene analyses were used to characterise the effect of rs560887 on G6PC2 pre-mRNA splicing. Fusion gene and gel retardation analyses characterised the effect of rs2232316 on G6PC2 promoter activity and transcription factor binding. The genetic association of rs2232316 with FPG variation was assessed using regression adjusted for age, sex and BMI in 4,220 Europeans with normal FPG. RESULTS: The rs560887-G allele was shown to enhance G6PC2 pre-mRNA splicing, whereas the rs2232316-A allele enhanced G6PC2 transcription by promoting Foxa2 binding. Genetic analyses provide evidence for association of the rs2232316-A allele with increased FPG (ß = 0.04 mmol/l; p = 4.3 × 10(-3)) as part of the same signal as rs560887, rs573225 and rs13431652. CONCLUSIONS/INTERPRETATION: As with rs13431652, the in situ functional data with rs560887 and rs2232316 are in accord with the putative function of G6PC2 in pancreatic islets, and suggest that all three are potentially causative SNPs that contribute to the association between G6PC2 and FPG.

Reassessment of the putative role of BLK-p.A71T loss-of-function mutation in MODY and type 2 diabetes.

AIMS/HYPOTHESIS: MODY is believed to be caused by at least 13 different genes. Five rare mutations at the BLK locus, including only one non-synonymous p.A71T variant, were reported to segregate with diabetes in three MODY families. The p.A71T mutation was shown to abolish the enhancing effect of BLK on insulin content and secretion from pancreatic beta cell lines. Here, we reassessed the contribution of BLK to MODY and tested the effect of BLK-p.A71T on type 2 diabetes risk and variations in related traits. METHODS: BLK was sequenced in 64 unelucidated MODY samples. The BLK-p.A71T variant was genotyped in a French type 2 diabetes case-control study including 4,901 cases and 4,280 controls, and in the DESIR (Data from an Epidemiological Study on the Insulin Resistance Syndrome) and SUVIMAX (Supplementation en Vitamines et Mineraux Antioxydants) population-based cohorts (n = 6,905). The variant effects were assessed by logistic and linear regression models. RESULTS: No rare non-synonymous BLK mutations were found in the MODY patients. The BLK p.A71T mutation was present in 52 normoglycaemic individuals, making it very unlikely that this loss-of-function mutation causes highly penetrant MODY. We found a nominal association between this variant and increased type 2 diabetes risk, with an enrichment of the mutation in the obese diabetic patients, although no significant association with BMI was identified. CONCLUSIONS/INTERPRETATION: No mutation in BLK was found in our MODY cohort. From our findings, the BLK-p.A71T mutation may weakly influence type 2 diabetes risk in the context of obesity; however, this will require further validation.

Contribution of 24 obesity-associated genetic variants to insulin resistance, pancreatic beta-cell function and type 2 diabetes risk in the French population.

CONTEXT: Obesity is the major determinant of type 2 diabetes (T2D), presumably through its effect on insulin resistance. Genome-wide association studies reported many single-nucleotide polymorphisms (SNPs) that increase obesity risk and body mass index (BMI), but their impact on T2D-related traits and risk is unclear. OBJECTIVE: We aimed at analyzing the effect of 24 obesity risk alleles, separately and in combination, on variation of both insulin resistance and ß-cell dysfunction, and on T2D risk. DESIGN: We genotyped 24 obesity-associated SNPs and calculated an obesity genotype score (sum of the obesity risk alleles per individual). We analyzed the contribution of each SNP and this score to the variation of four metabolic indices: homeostasis model assessment of insulin resistance (HOMA-IR), homeostasis model assessment of the pancreatic ß-cell function (HOMA-B), insulin sensitivity index (ISI) and insulinogenic index (II) (in up to 8050 nondiabetic French individuals) and to T2D risk (in 2077 T2D cases and 3085 controls). RESULTS: We found a highly significant effect of the obesity genotype score on increased insulin resistance adjusted for age and gender (ß=0.02; P-value=7.16 × 10(-9) for HOMA-IR). Individually, we identified nominal or significant association between increased insulin resistance and risk alleles in FAIM2, FTO, GNPDA2, MC4R, NPC1, PTER and SH2B1. Most signals, including the obesity genotype score and FTO SNP, were also associated with increased ß-cell function (ß=0.01; P-value=1.05 × 10(-6) and ß=0.04; P-value=3.45 × 10(-4), respectively). In our T2D case-control study, only the obesity genotype score and the well-known FTO locus significantly contributed to T2D risk (OR=1.03; P-value=9.99 × 10(-3) and OR=1.15; P-value=9.46 × 10(-4), respectively). Adjustment for BMI abolished all significant associations. CONCLUSIONS: Genetic predisposition to obesity contributes to increased insulin resistance and to its compensation through increased ß-cell function, and weakly increases the T2D risk. These associations are mediated by BMI.

Differential coexpression analysis of obesity-associated networks in human subcutaneous adipose tissue.

OBJECTIVE: To use a unique obesity-discordant sib-pair study design to combine differential expression analysis, expression quantitative trait loci (eQTLs) mapping and a coexpression regulatory network approach in subcutaneous human adipose tissue to identify genes relevant to the obese state. STUDY DESIGN: Genome-wide transcript expression in subcutaneous human adipose tissue was measured using Affymetrix U133 Plus 2.0 microarrays (Affymetrix, Santa Clara, CA, USA), and genome-wide genotyping data was obtained using an Applied Biosystems (Applied Biosystems; Life Technologies, Carlsbad, CA, USA) SNPlex linkage panel. SUBJECTS: A total of 154 Swedish families ascertained through an obese proband (body mass index (BMI) >30 kg m(-2)) with a discordant sibling (BMI>10 kg m(-2) less than proband). RESULTS: Approximately one-third of the transcripts were differentially expressed between lean and obese siblings. The cellular adhesion molecules (CAMs) KEGG grouping contained the largest number of differentially expressed genes under cis-acting genetic control. By using a novel approach to contrast CAMs coexpression networks between lean and obese siblings, a subset of differentially regulated genes was identified, with the previously GWAS obesity-associated neuronal growth regulator 1 (NEGR1) as a central hub. Independent analysis using mouse data demonstrated that this finding of NEGR1 is conserved across species. CONCLUSION: Our data suggest that in addition to its reported role in the brain, NEGR1 is also expressed in subcutaneous adipose tissue and acts as a central 'hub' in an obesity-related transcript network.

Design of a prospective, longitudinal cohort of people living with type 1 diabetes exploring factors associated with the residual cardiovascular risk and other diabetes-related complications: The SFDT1 study.

Type 1 diabetes mellitus (T1DM) is associated with a high risk of cardiovascular (CV) complications, even after controlling for traditional CV risk factors. Therefore, determinants of the residual increased CV morbidity and mortality remain to be discovered. This prospective cohort of people living with T1DM in France (SFDT1) will include adults and children aged over six years living with T1DM, recruited throughout metropolitan France and overseas French departments and territories. The primary objective is to better understand the parameters associated with CV complications in T1DM. Clinical data and biobank samples will be collected during routine visits every three years. Data from connected tools, including continuous glucose monitoring, will be available during the 10-year active follow-up. Patient-reported outcomes, psychological and socioeconomic information will also be collected either at visits or through web questionnaires accessible via the internet. Additionally, access to the national health data system (Health Data Hub) will provide information on healthcare and a passive 20-year medico-administrative follow-up. Using Health Data Hub, SFDT1 participants will be compared to non-diabetic individuals matched on age, gender, and residency area. The cohort is sponsored by the French-speaking Foundation for Diabetes Research (FFRD) and aims to include 15,000 participants.

Phytotherapies in motion: French Guiana as a case study for cross-cultural ethnobotanical hybridization.

BACKGROUND: French Guiana is characterized by a very multicultural population, made up of formerly settled groups (Amerindians, Maroons, Creoles) and more recent migrants (mostly from Latin America and the Caribbean). It is the ideal place to try to understand the influence of intercultural exchanges on the composition of medicinal floras and the evolution of phytotherapies under the effect of cross-culturalism. METHODS: A combination of qualitative and quantitative methods was used. Semi-directive interviews were conducted in 12 localities of French Guiana's coast between January 2016 and June 2017, and the responses to all closed questions collected during the survey were computerized in an Excel spreadsheet to facilitate quantitative processing. Herbarium vouchers were collected and deposited at the Cayenne Herbarium to determine Linnaean names of medicinal species mentioned by the interviewees. A list of indicator species for each cultural group considered was adapted from community ecology to this ethnobiological context, according to the Dufrêne-Legendre model, via the "labdsv" package and the "indval" function, after performing a redundancy analysis (RDA). RESULTS: A total of 205 people, belonging to 15 distinct cultural groups, were interviewed using semi-structured questionnaires. A total of 356 species (for 106 botanical families) were cited. We observed that pantropical and edible species hold a special place in these pharmacopeias. If compared to previous inventories, 31 recently introduced species can be counted. Furthermore, this study shows that the majority of the plants used are not specific to a particular group but shared by many communities. However, despite this obvious cross-culturalism of medicinal plants between the different cultural communities of French Guiana, divergent trends nevertheless appear through the importance of 29 indicator/cultural keystone species in 10 cultural groups. Finally, we have emphasized that the transmission of herbal medicine's knowledge in French Guiana is mainly feminine and intra-cultural. CONCLUSION: French Guianese medicinal flora is undoubtedly related to the multiple cultures that settled this territory through the last centuries. Cultural pharmacopeias are more hybrid than sometimes expected, but cultural keystone species nevertheless arise from a common background, allowing to understand, and define, the relationships between cultural groups.

EEG spectral power and sleepiness during 24 h of sustained wakefulness in patients with obstructive sleep apnea syndrome.

OBJECTIVE: This study investigated if obstructive sleep apnea syndrome (OSAS) may be associated with higher activity in different frequency bands of the EEG during a sustained wakefulness paradigm. METHODS: Twelve OSA patients and 8 healthy controls were studied with the Karolinska Drowsiness Test (KDT) and subjective ratings of sleepiness (VAS and KSS) conducted every hour during 24 h of sustained wakefulness. RESULTS: The waking EEG activity, mainly in the low (0.5-7.8 Hz) and fast (12.7-29.2 Hz) frequency band, increased as time awake progressed in both groups but more obviously in OSA patients. A similar pattern was observed for rated sleepiness in both groups. Moreover, VAS ratings of alertness were closely related to the awake theta, fast alpha and beta bands in controls but not in OSA patients. CONCLUSIONS: OSAS was associated with a wake-dependent increase in low (0.5-7.8 Hz) and fast (12.7-29.2 Hz) frequency range activity. Variations in behavioural sleepiness measured by VAS ratings closely reflect most of the waking EEG parameters in controls but not in OSA patients. SIGNIFICANCE: In a sustained wakefulness paradigm, higher activity in delta, theta and beta bands associated with OSAS indicates that OSA patients show marked signs of higher sleepiness and stronger efforts than controls to stay awake, even though they tend to underestimate their sleepiness.

Are OSAS patients impaired in their driving ability on a circuit with medium traffic density?

OBJECTIVES: Many studies have demonstrated that patients with Obstructive Sleep Apnea Syndrome (OSAS), a very common sleep-related breathing disorder, are usually impaired in their driving ability because of decreased sleep quality. However, most of the simulation procedures in laboratories are designed to create monotonic conditions with low traffic density, if any, thereby leading to a dramatic decrease in performance in OSAS patients because of the lack of stimulation. The aim of this study was therefore to evaluate driving abilities in OSAS patients involved in a driving simulation task with medium traffic density, in order to replicate as far as possible real world conditions. The behavioral and physiological attributes likely to predict driving performance in these patients were also investigated. METHODS: After a normal night of sleep, 12 OSAS patients and 8 healthy controls performed 6 driving sessions during a 24-h period of sustained wakefulness. Driving performances (speed, lateral position, distances...) were measured and correlated to sleep parameters and to a waking EEG recorded during the task. RESULTS: Compared to controls, patients showed difficulties in speed adjustment. However, they maintained longer inter-vehicle distances, including during overtaking. Their waking EEG, while driving, showed increased spectral power in theta (3.9-7.8Hz) but also in beta (12.7-29.2Hz) activity, alpha power (7.9-12.6Hz) being increased in both groups due to sustained wakefulness. Poor sleep indices were correlated to increased theta and beta activities, as well as to more cautious behavior. DISCUSSION: In medium traffic density conditions, driving performance in OSAS patients remained at near normal levels, but with more cautious behavior than controls. This could be the result of a bigger effort to stay awake, as suggested by an increased beta activity in these patients.

[Maintenance therapy of ulcer disease. Comparative multicenter study of sucralfate, cimetidine and a placebo]. / Traitement d'entretien de la maladie ulcéreuse. Etude multicentrique comparative du sucralfate, de la cimétidine et d'un placebo.

Relapse rates were studied in one hundred patients in a multicentric, randomized trial during and after maintenance therapy comparing sucralfate, cimetidine and placebo. These patients were previously treated by cimetidine for peptic ulcer and were considered cured after endoscopic examination. Outpatients were randomly assigned to a 6 month maintenance treatment with either cimetidine (600 mg daily), sucralfate (300 mg daily) or a placebo. All patients underwent endoscopic evaluation after 3 and 6 months of therapy. A clinical évaluation was performed 6 months after all treatment had ceased. Clinical and endoscopic results proved the significant superiority of both sucralfate and cimetidine over the placebo. Remission rates with sucralfate were respectively 80,4 p. 100 after 6 months and 68,5 p. 100 after 12 months. These results were slightly superior to those observed with cimetidine (69,3 p. 100 and 61,3 p. 100). However, this difference is not statistically significant. Results for the placebo group were 47,9 p. 100 and 37,7 p. 100. Sucralfate is an effective medication in preventing the recurrence of peptic ulcer. Its pharmacological action, its few side effects and its effectiveness seem to make this medication very interesting in treating the ulcerous disease.

Transcription factor gene MNX1 is a novel cause of permanent neonatal diabetes in a consanguineous family.

AIM: Permanent neonatal diabetes mellitus (PNDM) is a rare monogenic form of non-autoimmune diabetes. Genetic defects have been identified in∼60% of cases, with mutations in ABCC8, KCNJ11 and INS being the most frequent causes of PNDM. Recognition of genetic subtypes strongly impacts on both patients' care and family counseling. This study aimed to identify the genetic aetiology of PNDM in a diabetic girl born of consanguineous parents. METHODS: DNA samples from both the proband and her non-diabetic parents were analyzed for homozygosity mapping, using Illumina Infinium 660K SNP microarrays, focusing on the runs of homozygosity (ROHs) detected only in the patient. Standard Sanger sequencing of candidate genes (MNX1 and GATA6) present in the ROHs was subsequently performed, as well as expression analyses on human embryonic and adult pancreatic islet samples. RESULTS: A putative causal homozygous mutation in the transcription factor gene MNX1 (c.816C>A/p.Phe272Leu) was identified in the PNDM patient, who was clinically diagnosed as a typical case of PNDM with no developmental pancreatic defects or other clinical features. The probable deleterious mutation was located within the MNX1 homeodomain helix 2 that is highly conserved between species. In human embryonic pancreatic islet samples, it has been shown that MNX1 expression is significantly enriched in pancreatic epithelium compared with mesenchyme, suggesting a role for MNX1 in human pancreatic beta-cell development. CONCLUSION: This study found a new putative cause of PNDM in a consanguineous family. Replication in other cohorts would help to clarify the clinical spectrum of MNX1 mutations in PNDM patients.

Interaction of age with shift-related sleep-wakefulness, sleepiness, performance, and social life.

It is not clear how the age-related changes in sleep are related to performance and subjective sleepiness at different time of the day. The aim of the present study was to study work shift related interactions of age with sleep-wakefulness, performance, and social life. A representative sample of aircraft maintenance workers in a continuous three-shift system was studied by a questionnaire (n = 275) and an on-site field (n = 49) study. In the field study, sleep length and quality and different ratings of social and other activities were studied with an actigraphy and a Pocket PC diary during 15 consecutive days. Subjective sleepiness (KSS) and vigilance performance (PVT) were registered at work. Although the shift type influenced the sleep, subjective sleepiness, performance, and social life, age was distinctly related only to shift-related changes in the amount of sleep, subjective sleepiness, and psychomotor vigilance. Night shifts were related with shorter sleep, decreased performance, and increased sleepiness. Although subjective sleepiness was greatest among the youngest (25-34 years) age group during the morning and the night shifts, the increase of performance lapses was higher among the middle-aged (35-49 years) and senior (50-58 years) groups during the night shifts compared to the youngest age group. According to the questionnaire, older shiftworkers also tended to perceive more frequently that subjective sleepiness decreases their work performance during the morning and night shifts. The results indicate of no direct link between age-related differences in subjective sleepiness and performance at night work. The shorter day sleep after the night shifts and higher deterioration of subjective and objective performance according to age urge on development of shift schedules aiming at lower fatigue levels during the night shifts.

Development of gastric argyrophil carcinoid tumors in a case of Zollinger-Ellison syndrome with primary hyperparathyroidism during long-term antisecretory treatment.

Fundic argyrophil carcinoid tumors developed in the course of a 5-year continuous treatment with high dosages of H2-antagonists in a well-documented case of Zollinger-Ellison syndrome with primary hyperparathyroidism, high basal acid output, and serum gastrin. Approximately 100 small polyps were disseminated throughout the gastric fundus exclusively, leading to total gastrectomy. Metastatic carcinoid in a lymph node and pancreatic gastrinomas also were found at surgery. Gastric endocrine cell proliferation varied from simple argyrophil cell hyperplasia to carcinoid tumors eroding the surface and infiltrating the submucosa. Ultrastructural studies showed that the tumoral proliferation was heterogeneous, and included tumors composed of enterochromaffin (EC) and typical enterochromaffin-like (EC-L) cells, and tumors in which a majority of cells exhibited dense round granules resembling those of A-like or D1/P endocrine cell types. The risk of developing gastric fundic carcinoid tumors in ZES patients submitted to long-term antisecretory treatment should be given increased attention.