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Background: Gestational diabetes mellitus (GDM) contributes significantly to maternal and neonatal morbidity, but data from marginalized populations remains scarce. This study aims to compare risk-factor-based screening to universal testing for GDM among migrants along the Thailand-Myanmar border. Methods: From the prospective cohort (September 2016, February 2019), 374 healthy pregnant women completed a 75g oral glucose tolerance test (OGTT) at 24-32 weeks gestation. Fasting, one hour and two hour cut-offs were based on Hyperglycaemia and Adverse Pregnancy Outcomes (HAPO trial) criteria and cases were treated. The sensitivity and specificity of risk-factor-based screening criteria was calculated using OGTT as the gold standard. Risk factors included at least one positive finding among 10 criteria, e.g., obesity (body mass index (BMI) ≥27.5kg/m2), 1st degree relative with diabetes etc. Adverse maternal and neonatal outcomes were compared by GDM status, and risk factors for GDM were explored. Results: GDM prevalence was 13.4% (50/374) (95% CI: 10.3-17.2). Risk-factors alone correctly identified 74.0% (37/50) OGTT positive cases: sensitivity 74.0% (59.7-85.4) and specificity 27.8% (3.0-33.0). Burman women accounted for 29.1% of the cohort population, but 38.0% of GDM cases. Percentiles for birthweight (p=0.004), head circumference (p=0.005), and weight-length ratio (p=0.010) were higher in newborns of GDM mothers compared with non-GDM, yet 21.7% (75/346) of newborns in the cohort were small-for-gestational age. In Burman women, overweight/obese BMI was associated with a significantly increased adjusted odds ratio 5.03 (95% CI: 1.43-17.64) for GDM compared to normal weight, whereas underweight and overweight/obese in Karen women were both associated with similarly elevated adjusted odds, approximately 2.4-fold (non-significant) for GDM. GDM diagnosis by OGTT was highest prior to peak rainfall. Conclusions: Risk-factor-based screening was not sufficiently sensitive or specific to be useful to diagnose GDM in this setting among a cohort of low-risk pregnant women. A two-step universal screening program has thus been implemented.

Autoimmune diseases comprise a wide group of diseases involving a self-response of the immune system against the host. The etiopathogenesis is very complex involving disease-specific factors but also environmental factors, among which the diet. Maternal diet during pregnancy as well as early nutrition recently attracted the interest of the scientists as contributing to the immune programming. In this paper, we reviewed the most recent literature on the effect of maternal diet and early nutrition in modulating the immune system in a selected subset of autoimmune diseases: type 1 diabetes, celiac disease, inflammatory bowel disease, juvenile idiopathic arthritis and rheumatoid arthritis. Particularly, we focused our narrative on the role of maternal and perinatal nutrition in the epigenetic mechanisms underlying the auto-immune response. Maternal diet during pregnancy as well as breastfeeding and early nutrition play a big role in many epigenetic mechanisms. Most of the nutrients consumed by the mother and the infant are known exerting epigenetic functions, such as folate, methionine, zinc, vitamins B12 and D, fibers, casein and gliadin, and they were linked to gene expression changes in the immune pathways. Despite the common role of maternal diet, breastfeeding and early nutrition in almost all the autoimmune diseases, each disease seems to have specific diet-driver epigenetic mechanisms that require further investigations. The research in this field is opening new routes to establishing a precision nutrition approach to the auto-immune diseases.

In Qatar, Type 1 Diabetes mellitus (T1DM) is one of the most prevalent disorders. This study aimed to explore the gut microbiome’s relation to the continuous subcutaneous insulin infusion (CSII) therapy, dietary habits, and the HbA1c level in the pediatric T1DM subjects in Qatar. We recruited 28 T1DM subjects with an average age of 10.5 ± 3.53 years. The stool sample was used to measure microbial composition by 16s rDNA sequencing method. The results have revealed that the subjects who had undergone CSII therapy had increased microbial diversity and genus Akkermansia was significantly enriched in the subjects without CSII therapy. Moreover, genus Akkermansia was higher in the subjects with poor glycemic control (HbA1c &gt, 7.5%). When we classified the subjects based on dietary patterns and nationality, Akkermansia was significantly enriched in Qataris subjects without the CSII therapy consuming Arabic diet than expatriates living in Qatar and eating a Western/mixed diet. Thus, this pilot study showed that abundance of Akkermansia is dependent on the Arabic diet only in poorly controlled Qataris T1DM patients, opening new routes to personalized treatment for T1DM in Qataris pediatric subjects. Further comprehensive studies on the relation between the Arabic diet, ethnicity, and Akkermansia are warranted to confirm this preliminary finding.

Chronic kidney disease (CKD) is a globally common and important disease and there are evidence for a bidirectional relationship between microbiota and CKD. The aim of the study was to examine the influence of prebiotic – gum acacia (GA) on the intestinal microbiota in rats with adenine-induced CKD. Animals were randomly distributed into four equal groups (n = 6): control, adenine, GA and adenine + GA groups. CKD was induced by adenine (0.75% w/w) given in the diet daily for four weeks, and GA was administered in drinking water at a concentration of 15% w/v. The 16s rRNA analysis was performed on Illumina Miseq targeting V3-V4 region to characterize microbial composition. The abundance of Actinobacteria, Proteobacteria, Tenericutes and Verrucomicrobia bacteria was increased in adenine-induced CKD, and GA treatment successfully reversed those levels. Interestingly, alpha and beta diversity index were both reduced with GA treatment in rats with CKD. Short chain fatty acids (SCFAs) measurement and PICRUSt analysis have shown that GA treatment completely restored the depleted butyrate level and various perturbated functional pathways, respectively, in CKD rats. Taking together, our results suggest that GA supplementation has a beneficial role in treating CKD, through an increased production of butyrate, as well as its anti-inflammatory, antioxidant capacity and anti-nitrosative properties.

Type 1 diabetes (T1D) appears most frequently in childhood, with an alarming increasing incidence in the last decades. Although the genetic predisposition is a major risk factor, it cannot solely explain the complex etiology of T1D which is still not fully understood. In this paper, we reviewed the most recent findings on the role of early nutrition and the involvement of the gut microbiota in the etiopathogenesis of T1D. The main conclusions that are withdrawn from the current literature regarding alleviating the risk of developing T1D through nutrition are the encouragement of long-term breast-feeding for at least the first 6 months of life and the avoidance of early complementary foods and gluten introduction (before 4 months of age) as well as cow milk introduction before 12 months of life. These detrimental feeding habits create a gut microbiota dysbiotic state that can contribute to the onset of T1D in infancy. Finally, we discussed the possibility to introduce probiotics, prebiotics and post-biotics in the prevention of T1D.

Chronic kidney disease (CKD) is a globally common and important disease and there are evidence for a bidirectional relationship between microbiota and CKD. The aim of the study was to examine the influence of prebiotic - gum acacia (GA) on the intestinal microbiota in rats with adenine-induced CKD. Animals were randomly distributed into four equal groups (n = 6): control, adenine, GA and adenine + GA groups. CKD was induced by adenine (0.75% w/w) given in the diet daily for four weeks, and GA was administered in drinking water at a concentration of 15% w/v. The 16s rRNA analysis was performed on Illumina Miseq targeting V3-V4 region to characterize microbial composition. The abundance of Actinobacteria, Proteobacteria, Tenericutes and Verrucomicrobia bacteria was increased in adenine-induced CKD, and GA treatment successfully reversed those levels. Interestingly, alpha and beta diversity index were both reduced with GA treatment in rats with CKD. Short chain fatty acids (SCFAs) measurement and PICRUSt analysis have shown that GA treatment completely restored the depleted butyrate level and various perturbated functional pathways, respectively, in CKD rats. Taking together, our results suggest that GA supplementation has a beneficial role in treating CKD, through an increased production of butyrate, as well as its anti-inflammatory, antioxidant capacity and anti-nitrosative properties.

The expression of Annexin A3 (ANXA3), a member of a family of calcium‐binding proteins, is restricted to neutrophils. Its abundance is increased in septic patients, yet the functional relevance of ANXA3 in this context remains unknown. Upon reviewing evidence from public transcriptome repositories and the literature, we hypothesize that during sepsis ANXA3 could mediate neutrophils microbicidal activity while also prolonging their survival, and thus also possibly contributing to organ damage., Summary According to publicly available transcriptome datasets, the abundance of Annexin A3 (ANXA3) is robustly increased during the course of sepsis; however, no studies have examined the biological significance or clinical relevance of ANXA3 in this pathology. Here we explored this interpretation gap and identified possible directions for future research. Based on reference transcriptome datasets, we found that ANXA3 expression is restricted to neutrophils, is upregulated in vitro after exposure to plasma obtained from septic patients, and is associated with adverse clinical outcomes. Secondly, an increase in ANXA3 transcript abundance was also observed in vivo, in the blood of septic patients in multiple independent studies. ANXA3 is known to mediate calcium‐dependent granules–phagosome fusion in support of microbicidal activity in neutrophils. More recent work has also shown that ANXA3 enhances proliferation and survival of tumour cells via a Caspase‐3‐dependent mechanism. And this same molecule is also known to play a critical role in regulation of apoptotic events in neutrophils. Thus, we posit that during sepsis ANXA3 might either play a beneficial role, by facilitating microbial clearance and resolution of the infection; or a detrimental role, by prolonging neutrophil survival, which is known to contribute to sepsis‐mediated organ damage.

PurposeA successful pregnancy relies on the interplay of various biological systems. Deviations from the norm within a system or intersystemic interactions may result in pregnancy-associated complications and adverse pregnancy outcomes. Systems biology approaches provide an avenue of unbiased, in-depth phenotyping in health and disease. The molecular signature in pregnancy (MSP) cohort was established to characterise longitudinal, cross-omic trajectories in pregnant women from a resource constrained setting. Downstream analysis will focus on characterising physiological perturbations in uneventful pregnancies, pregnancy-associated complications and adverse outcomes.ParticipantsFirst trimester pregnant women of Karen or Burman ethnicity were followed prospectively throughout pregnancy, at delivery and until 3 months post partum. Serial high-frequency sampling to assess whole blood transcriptomics and microbiome composition of the gut, vagina and oral cavity, in conjunction with assessment of gene expression and microbial colonisation of gestational tissue, was done for all cohort participants.Findings to date381 women with live born singletons averaged 16 (IQR 15–18) antenatal visits (13 094 biological samples were collected). At 5% (19/381) the preterm birth rate was low. Other adverse events such as maternal febrile illness 7.1% (27/381), gestational diabetes 13.1% (50/381), maternal anaemia 16.3% (62/381), maternal underweight 19.2% (73/381) and a neonate born small for gestational age 20.2% (77/381) were more often observed than preterm birth.Future plansResults from the MSP cohort will enable in-depth characterisation of cross-omic molecular trajectories in pregnancies from a population in a resource-constrained setting. Moreover, pregnancy-associated complications and unfavourable pregnancy outcomes will be investigated at the same granular level, with a particular focus on population relevant needs such as effect of tropical infections on pregnancy. More detailed knowledge on multiomic perturbations will ideally result in the development of diagnostic tools and ultimately lead to targeted interventions that may disproportionally benefit pregnant women from this resource-limited population.Trial registration numberNCT02797327.

Background Chronic kidney disease (CKD) patients under hemodialysis show a higher risk of cardiovascular (CV) mortality and morbidity than the general population. This study aims to identify genetic markers that could explain the increased CV risk in hemodialysis. Methods A total of 245 CKD patients under hemodialysis were recruited and followed up for 5 years to record CV events. Genetic analysis was performed using single-nucleotide polymorphisms (SNPs) genotyping by Infinium Expanded Multi-Ethnic Genotyping Array (Illumina, San Diego, CA, USA) comparing patients with and without a history of CV events [161 cardiovascular diseases (CVDs) and 84 no CVDs]. The fixation index (Fst) measure was used to identify the most differentiated SNPs, and gene ontology analysis [Protein Analysis THrough Evolutionary Relationships (PANTHER) and Ingenuity Pathway Analysis (IPA)] was applied to define the biological/pathological roles of the associated SNPs. Partitioning tree analysis interrogated the genotype–phenotype relationship between discovered genetic variants and CV phenotypes. Cox regression analysis measured the effect of these SNPs on new CV events during the follow-up (FU). Results Fst analysis identified 3218 SNPs that were significantly different between CVD and no CVD. Gene ontology analysis identified two of these SNPs as involved in cardiovascular disease pathways (Ingenuity Pathway) and heart development (Panther) and belonging to 2 different genes: Glucagon-like peptide-1 receptor (GLP1R) and Sarcoglycan delta (SGCD). The phenotype–genotype analysis found a higher percentage of CVD patients carrying the GLP1R rs10305445 allele A (P = 0.03) and lower percentages of CVD patients carrying the SGCD rs145292439 allele A (P = 0.038). Moreover, SGCD rs145292439 was associated with higher levels of high-density lipoprotein (P = 0.015). Cox analysis confirmed the increased frequency of CV events during the 5-year FU in patients carrying GLP1R rs1035445 allele A but it did not show any significant association with SGCD rs145292439. Conclusions This study identified GLP1R rs10305445 and SCGD rs145292439 as potential genetic markers that may explain the higher risk of CVD in hemodialysis patients.

This study focused on Qatar’s pediatric population that has witnessed a steep increase in the incidence of the disease. In order to understand this, we analyzed the blood and stool samples of a pilot group of 21 T1D subjects (age 6-12 years old) for the microbiome composition, Short Chain Fatty Acid (SCFA) levels and methylation profiles using 16s rDNA sequencing, gas Chromatography and Infinium methylation assay respectively.

IntroductionGestational diabetes mellitus (GDM) affects 23.6% of Qatari women and is associated with maternal and perinatal morbidity and long-term risk of developing type 2 diabetes. A number of challenges exist with current interventions, including non-compliance with dietary advice, the reluctance of mothers to ingest metformin tablets or use insulin injections. These challenges highlight the importance of pursuing evidence-based prevention strategies. Myo-inositol is readily available as an US Food and Drug Administration-approved food supplement with emerging but limited evidence suggesting it may be beneficial in reducing the incidence of GDM. Further studies, such as this one, from different ethnic contexts and with differing risk factors, are urgently needed to assess myo-inositol effects on maternal and neonatal outcomes.Methods and analysisThis study is a prospective, randomised, double-blinded, placebo controlled clinical trial to either myo-inositol supplementation or placebo.We plan to enrol 640 pregnant women attending antenatal care at Sidra Medicine, Doha, Qatar, 320 in each arm. All participants will complete at least 12 weeks of supplementation prior to undertaking the Oral Glucose Tolerance Test at 24–28 weeks. The daily use of the trial supplementation will continue until the end of pregnancy. All outcome measures will be collected from the electronic medical records.Ethics and disseminationEthical approval for the study was obtained on 12 April 2021 from Sidra Medicine (IRB number 1538656). Results of the primary trial outcome and secondary endpoints will be submitted for publication in a peer-reviewed journal.Trial registration numberProspectively registered on 26 May 2021. Registration number ISRCTN16448440 (ISRCTN registry).

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Background: The composition of the microbiome in human body sites plays an important role in health. The vaginal environment is colonized by several species of bacteria that have a major influence on reproductive health. The advancement of sequencing technologies has made the assessment of the composition of the microbiota possible through microbial DNA extraction and sequencing. Therefore, it is of a paramount importance to select a sensitive and reproducible DNA extraction method, that facilitates isolation of microbial DNA with a sufficient quantity and purity, from microbial species living in the vaginal environment. Here, we have evaluated four different DNA extraction protocols from self-collected vaginal swabs. Methods: Five healthy female volunteers were enrolled in the study. Each donor was asked to self-collect 4 samples using Copan ESwab. DNA was extracted using Qiagen DNeasy kit and three modified protocols of the MoBio PowerSoil kit (“DNeasy PowerSoil” after acquisition from Qiagen). DNA quantity and integrity was checked through Nanodrop and LabChip GX. DNA was further tested through quantitative real-time PCR (qPCR) and 16S sequencing. Vaginal microbiota diversities were determined using MiSeq-Illumina high-throughput sequencing of bacterial 16S rDNA V1–V3 fingerprint. Sequencing data were analyzed using QIIME pipeline. Results: Qiagen DNeasy protocol resulted in the highest DNA yield as compared to the modified protocols of MoBio Powersoil kit. The size of the DNA extracted using each protocol was ~40 kb. Qiagen DNeasy protocol gave the highest Genomic Quality Score (average ± standard deviation: 4.24 ± 0.36), followed by the different MoBio Powersoil protocols. A substantial variability in microbial DNA abundance was found across the protocols. The vaginal microbiota of the healthy volunteers was dominated by Lactobacillus species. MoBio Powersoil kit provided a significantly higher alpha diversity as compared to the Qiagen DNeasy kit, while beta diversity measures did not reveal any significant cluster changes, except when the Bray-Curtis method was applied. Conclusion: We were able to isolate microbial DNA from the vaginal swabs. Qiagen DNeasy method gave the highest DNA yield and quality but was not optimal in detecting microbial diversity. The modified MoBio PowerSoil protocols showed higher microbial diversities as compared to the standard protocol.

Steps involved in reductionist investigation approaches can be imitated using public transcriptome datasets as source of training material. In the present report trainees explored an apparent gap in biological knowledge for FAM129A (family with sequence similarity 129 member A). Elevated abundance of FAM129A transcripts were observed in a transcriptome dataset where neutrophils were exposed in vitro to plasma of patients with sepsis. However, no literature linking FAM129A and either neutrophils, sepsis or inflammation could be identified. Additional datasets were selected to independently validate this initial observation and further explore differential expression of FAM129A in the context of sepsis studies. Follow on investigations carried out at the bench confirmed restriction of the expression of FAM129A protein at the surface of circulating blood neutrophils and monocytes. A potential role for FAM129A in neutrophil survival was inferred from profiling of literature associated with FAM129A, which remains to be investigated in further follow on investigations.

IntroductionPreterm birth (PTB) results from heterogeneous influences and is a major contributor to neonatal mortality and morbidity that continues to have adverse effects on infants beyond the neonatal period. This protocol describes the procedures to determine molecular signatures predictive of PTB through high-frequency sampling during pregnancy, at delivery and the postpartum period.Methods and analysisFour hundred first trimester pregnant women from either Myanmar or Thailand of either Karen or Burman ethnicity, with a viable, singleton pregnancy will be enrolled in this non-interventional, prospective pregnancy birth cohort study and will be followed through to the postpartum period. Fortnightly finger prick capillary blood sampling will allow the monitoring of genome-wide transcript abundance in whole blood. Collection of stool samples and vaginal swabs each trimester, at delivery and postpartum will allow monitoring of intestinal and vaginal microbial composition. In a nested case–control analysis, perturbations of transcript abundance in capillary blood as well as longitudinal changes of the gut, vaginal and oral microbiome will be compared between mothers giving birth to preterm and matched cases giving birth to term neonates. Placenta tissue of preterm and term neonates will be used to determine bacterial colonisation as well as for the establishment of coding and non-coding RNA profiles. In addition, RNA profiles of circulating, non-coding RNA in cord blood serum will be compared with those of maternal peripheral blood serum at time of delivery.Ethics and disseminationThis research protocol that aims to detect perturbations in molecular trajectories preceding adverse pregnancy outcomes was approved by the ethics committee of the Faculty of Tropical Medicine, Mahidol University in Bangkok, Thailand (Ethics Reference: TMEC 15–062), the Oxford Tropical Research Ethics Committee (Ethics Reference: OxTREC: 33–15) and the local Tak Province Community Ethics Advisory Board. The results of this cooperative project will be disseminated in multiple publications staggered over time in international peer-reviewed scientific journals.Trial registration numberNCT02797327; Pre-results.

Introduction:The human body harbors bacteria, archaea, viruses, and eukaryotic microbes that inhabit interactive interfaces exposed to the external environment. This collection of microorganisms consists of about 100 trillion of microbes called the human microbiota and the total genes encoded by these microbes collectively called as human microbiome. They are strongly associated with the development of non-communicable diseases (NCDs) in addition to the role they play in communicable infectious diseases. Changes in microbiota has been reported in various NCDs including obesity, cancer, diabetes, metabolic syndrome, inflammatory bowel disease (IBD), asthma, cardiovascular disease (CVD), kidney disease (KD) and others. Advances in DNA sequencing and bioinformatics had enabled researchers the exploration of the genetic diversity of the uncultured component of host-associated microbial communities. The aim of this work is to test different DNA extracting protocols to characterize human microbiota of biological samples collected from adult volunteers. Materials & Methods: Biological samples such as human saliva, vaginal swabs and stool were collected from well-characterized adult participants. Total DNA were extracted from Vaginal swabs (n = 5) using DNeasy Blood & Tissue Kit and Mobio power Soil protocol; Stool samples (n = 17) using MoBio Power Fecal and Qiamp fast stool kits; Saliva samples (n = 8) using QIASymphony DNA Midi kit. DNA quantity and quality was assessed by Nanodrop spectrophotometer. Integrity of the DNA was reviewed on LabChip. Human microbiota diversities of different biological samples through different extraction protocols will be determined using MiSeq-Illumina high-throughput sequencing of bacterial 16S rDNA V1-V3 fingerprint. Sequenced data will be analyzed using QIIME pipeline. Results DNA measurement revealed that DNeasy Blood & Tissue Kit gave higher DNA yield with good integrity (2-8 ng/μL) than Mobio power Soil protocol (0.6-2 ng/μL) for Vaginal swabs. Likewise, MoBio Power Fecal kit (1.8-41.5 ng/mL) gave higher DNA yield than Qiamp fast stool kit (5-21 ng/mL) for stool samples. On the other hand, DNA Qiasymphony DSP kit gave (3-241 ng/μL) for saliva. Expected Outcome: From the previous studies by other groups, it is expected that Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria are the most abundant phyla in human microbiota. The abundance of the mentioned phylum may differ with respect to body sites. Actinobacteria members will be abundant on skin, Firmicutes and Bacteroidetes will be more abundant in gut. Firmicutes, Proteobacteria, and Bacteroidetes will be the more abundant in saliva and Firmicutes in Saliva. In addition, Each DNA extraction protocol has different strategies to lyse the human microbes. We speculate that the microbial diversity of same body site can be different with different protocols. Characterization of healthy human microbiome gives a better understanding of their role in human health during diseased conditions. Especially, Human gut microbiota influence the metabolism of host and it links to several metabolic disorders such as obesity, Type 2 diabetes and Metabolic syndrome when there is an imbalance in microbiota. Exploring Vaginal microbiome can guide as to find new therapeutic targets to treat several infections and to treat complications with preterm birth to pregnant women. This work will be a promising candidate to choose the adequate protocol based on the nature of the sample, which can guide us to explore the human microbiome in right path.

Background/Objective: Qatar suffers the highest percentage of non-communicable diseases (NCD), such as diabetes and obesity among few countries in the world. There are two major components that could explain the NCD, which are diet and the genetic background. It's well demonstrated that diet affects the genome function in terms of epigenetic and microbiome modification. Recent scientific advancement or studies have shown that all these components could be the factors in defining the development of the NCD, but, there is no study that explains the direct connection of Diet-Microbiome-Epigenome. We want to explore the interaction of these three factors in a small pilot study on type 1 diabetes (T1D) creating a methodology that will be applicable for all NCDs and nutrition-related diseases. This study aims to identify specific nutrients that modulate gut microbiome; to define different microbiome composition and metabolites; and to define differential methylated regions (DMR) that is possibly affected by nutrients, gut microbiome and its metabolite. This abstract presents the study design and discusses methodologies that will be used in the study. Study design and methodology: We will compare 4 groups of subjects: T1D, T1D obese, pure obese, and lean controls. Pediatric patients will be recruited from Sidra OPC based on major inclusion criteria, (such as age should be between 6-12 yrs, no antibiotic treatment in the past 3 months, no chronic diseases except T1D and no history of cancer) and divided into four groups: (1) healthy lean children (5-84th percentile of BMI), (2) Obese ( ≥ 95th percentile of BMI;), (3) T1D and (4) Obese T1D. A comprehensive set of physical measurements (body weight, height and Waist circumference), clinical biomarkers for diabetes and obesity (mainly, blood glucose level, lipid and liver profile, HbA1c will be done with blood sample) and family history of diabetes, treatment history will be collected and most importantly dietary habits will be collected by 24 hrs food recall. Two sets of stool samples (one for microbiome analysis by 16S rDNA-sequencing; and one for fatty acids analysis by gas chromatography) and blood samples (one for DNA extraction for methylation analysis by Illumina DNA-methylation Array and one for RNA extraction for gene expression analysis using Fluidigm platform) will be collected from each subject. Data analysis will investigate any association of diet to the clinical phenotypes comparing the four groups of subjects, using logistic regression; two-sided P-value of

The cells of the human body have a receptor that is sensitive to extracellular calcium [calcium-sensing receptor (CaSR)] and whose tissue expression is decreased in patients with chronic kidney disease. The CaSR plays a key role both in the regulation of PTH secretion by parathyroid cells, as well as in calcium tubular reabsorption by renal tubular cells. Recent biochemical and cellular studies found that this receptor has peculiar functional characteristics. Furthermore, pharmacological research has brought to the marker agonists for CaSR (calcimimetics) that today are used for treating secondary hyperparathyroidism and may have an effect on cardiovascular and bone diseases in patients with chronic kidney disease. With regard to the latter, the EVOLVE and ADVANCE trials were dedicated to understanding the role of cinacalcet, a calcimimetic drug, in the prevention of vascular calcification and cardiovascular events in patients with chronic kidney disease. Research in this field has thus opened promising p...

Parathyroid tumors display reduced sensitivity to extracellular calcium ([Ca2+]o). [Ca2+]o activates calcium-sensing receptor (CASR), which interacts with the scaffold protein filamin A (FLNA). The study aimed to investigate: (1) the FLNA expression in human parathyroid tumors, (2) its effects on the CASR mRNA and protein expression, and (3) on ERK signaling activation, (4) the effect of the carboxy-terminal CASR variants and (5) of the treatment with the CASR agonist R568 on FLNA-mediated ERK phosphorylation in HEK293 cells. Full-length FLNA immunostaining was variably reduced in parathyroid tumors. Immunofluorescence showed that FLNA localized in membrane and cytoplasm and co-localized with CASR in parathyroid adenomas (PAds)-derived cells. Cleaved C-terminus FLNA fragment could also be detected in PAds nuclear protein fractions. In HEK293 cells transfected with 990R-CASR or 990G-CASR variants, silencing of endogenous FLNA reduced CASR mRNA levels and total and membrane-associated CASR proteins. In agreement, FLNA mRNA levels positively correlated with CASR expression in a series of 74 PAds; however, any significant correlation with primary hyperparathyroidism severity could be detected and FLNA transcript levels did not differ between PAds harboring 990R or 990G CASR variants. R568 treatment was efficient in restoring 990R-CASR and 990G-CASR sensitivity to [Ca2+]o in the absence of FLNA. In conclusion, FLNA is downregulated in parathyroid tumors and parallels the CASR expression levels. Loss of FLNA reduces CASR mRNA and protein expression levels and the CASR-induced ERK phosphorylation. FLNA is involved in receptor expression, membrane localization and ERK signaling activation of both 990R and 990G CASR variants.

Background Paraoxonase 1 (PON1) gene polymorphisms and polyphenols intake have been reported independently associated to lipid profile and susceptibility to atherosclerosis and cardiovascular disease. However, the interaction between these factors remains to be investigated. We performed an observational nutrigenetic study to examine whether the interaction between polyphenols and anthocyanins intake and PON1 genetic variants can modulate biomarkers of cardiovascular health in an Italian healthy population. Methods We recruited 443 healthy volunteers who participated in the EC funded ATHENA project (AnThocyanin and polyphenols bioactive for Health Enhancement through Nutritional Advancement). Data collection included detailed demographic, clinical, dietary, lifestyle, biochemical and genetic data. Polyphenols and anthocyanins intake was measured by 24 h dietary recall repeated three times a year in order to get seasonal variations. We tested the interaction between 18 independent tagging SNPs in PON1 gene and polyphenols intake on HDL, LDL, cholesterol, triglycerides and atherogenic index of plasma. Results Without considering the genetic background, we could not observe significant differences in the lipid profile between high and low polyphenols and anthocyanins intake. Using a nutrigenetic approach, we identified protective genotypes in four independent polymorphisms that, at Bonferroni level (p ≤ 0.0028), present a significant association with increased HDL level under high polyphenols and anthocyanins intake, compared to risk genotypes (rs854549, Beta = 4.7 per C allele; rs854552, Beta = 5.6 per C allele; rs854571, Beta = 3.92 per T allele; rs854572, Beta = 3.94 per C allele). Conclusions We highlight the protective role of genetic variants in PON1 towards cardiovascular risk under high polyphenols and anthocyanins consumption. PON1 variants could represent novel biomarkers to stratify individuals who might benefit from targeted dietary recommendation for health promotion and strategies of preventive medicine. Electronic supplementary material The online version of this article (doi:10.1186/s12967-016-0941-6) contains supplementary material, which is available to authorized users.

Obesity prevalence has been growing exponentially over the last few decades, with a high impact in high-income countries, like Qatar. Several approaches are attempting to understand the causes of this phenomenon however more important is what to do to reverse the trends. Obesity is widely studied, mostly in Europe and the Unites States, and a number of studies have demonstrate the role of specific gene patterns, transcriptome and proteome pathways, and gut microbiome strains. The Omics sciences have a great potential to investigate the determinants of non-communicable diseases, such as obesity. Nutritional genomics sciences apply all the Omics approaches to address nutrition-related diseases, investigating the interaction between genes and diet. To date, few data are available from nutrigenomic studies conducted in Middle East and particularly in Qatar to help the design of targeted interventions. The high incidence of obesity and the peculiar genetic make-up of the Qatari population provide opportunities for exploring nutrigenomic approaches to help addressing the problem.

BackgroundVascular calcification and increased mortality is now well linked in hemodialysis (HD) patients. Among the inhibitors and promoters of calcium-phosphate and bone metabolism, we studied Sclerostin, a glycoprotein mainly expressed by osteocytes, involved in regulating bone formation by inhibiting Wnt–β-catenin signaling. Studies demonstrated, in experimental CDK animal model, a positive association between Sclerostin production and dietary phosphate intake.Materials and MethodsWe investigated the relation between Sclerostin levels (SL) and dietary habits in 49 Caucasian HD patients (age 36–90, M/F 31/18, dialysis vintage 1–144 months). Nutrient intake was analyzed with 24 h-recall method. Serum Sclerostin was measured by ELISA method. Statistically analysis was performed by SPSS software.ResultsMean calcium and phosphate intake were 705 ± 584 and 1196 ± 498 mg/day, respectively. Serum SL (3356 ± 2118 pg/ml) were increased in HD patients. Positive correlation was found between serum SL and calcium (r = 0.33, p = 0.025) and phosphate intake (r = 0.34, p = 0.021). Patients with SL higher were older (p = 0.003), showed higher protein (p = 0.012), calcium (p = 0.017) and phosphate (p = 0.004) intake. Multiple linear regression testing Sclerostin as dependent variable (dialysis vintage, age, body-weight, serum calcium and phosphate, serum PTH, calcium and phosphate intake as independent variables) found that dietary phosphate intake was the only significant determinant of SL (r = 0.356, B = 1.5, p = 0.004). In Patients with calcium carbonate or acetate therapy (n = 31), serum SL was positively correlated with dietary calcium (r = 0.433, p = 0.017) and phosphate intake (r = 0.377, p = 0.04).ConclusionsThe dietary phosphate and calcium intake may influence serum Sclerostin levels and potentially affect bone remodeling or soft tissue calcification in HD patients.

A population of 503 adult Caucasians was recruited to evaluate the effect of diet on cardiovascular risk factors and its interaction with the genetic background. Nutritional data were collected by 24h recalls and Mediterranean diet adherence was investigated by QuADM-15 questionnaire. Anthropometric measurements (weight, height, and waist circumference), lipid profile, serum glucose, electrolytes, homocysteine and hpCRP were measured. The genome-wide tag-SNPs analysis was performed by Illumina 200K SNPs array, enriched by the imputation analysis. The Gene*Environment (G*E) analysis evaluated candidate genes and SNPs for the phenotypes HDL and LDL, and the interaction with diet (omnivores vs. vegetarians) and anthocyanins intake.The dietary habit analysis identified 120 vegetarians and 374 omnivores. The distribution of macronutrients showed, as expected, a higher intake of animal proteins, saturated fatty acids and lower fiber intake in the omnivores. The consumption of anthocyanins and polyphenols was low in the entire population. Correlations were found between diet habits (omnivores vs vegetarians) and BMI (p

BackgroundParathyroid tumors display reduced sensitivity to extracellular calcium ([Ca2+]o). The parathyroid cell sensitivity to [Ca2+]o is mediated by the calcium-sensing receptor (CASR), a G-protein-coupled receptor interacting with the scaffold protein filamin A (FLNA). We investigated: the FLNA expression in human parathyroid tumors, its effect on the CASR membrane stabilization, and on ERK signaling activation in HEK293 cells, the effect of the C-terminal CASR variants (R990G) on the interaction with FLNA.MethodsFLNA expression was analyzed by immunohistochemistry and immunofluorescence in parathyroid tumors and in cells from parathyroid adenomas (PAds). Endogenous FLNA in HEK293 cells transfected with the CASR variants was silenced by siRNA technique. Results FLNA expression was down-regulated in human parathyroid tumors; In 74 PAds, FLNA mRNA levels positively correlated with CASR levels. In HEK293 cells transfected with 990R-CASR or 990G-CASR, FLNA silencing reduced both the total and membrane protein expression levels. However, in presence of endogenous FLNA, 990G-CASR expression in cell membrane was higher than that of the 990R-CASR. FLNA loss reduced significantly p-ERK levels induced both 990R-CaSR and 990G-CaSR. Treatment with the CASR agonist R568 delete the effect of FLNA loss and restoring 990G-CASR sensitivity to [Ca2+]o in absence of FLNA.ConclusionsFLNA is down-regulated in parathyroid tumors and paralleled the CASR expression levels. Loss of FLNA reduced the CASR expression levels and the CASR-induced ERK phosphorylation. The CASR 990G allele was associated with increased sensitivity to [Ca2+]o, though FLNA was required to determine the 990G-CASR higher expression and activity than that of 990R-CASR protein.

BackgroundAutosomal Dominant Hypocalcemia (ADH) is an endocrine disorder due to activating mutations of the calcium-sensing receptor (CASR) gene that encodes for a plasma membrane G protein-coupled receptor. This protein plays a central role in maintaining calcium homeostasis. ADH is characterized by hypocalcemia and hypercalciuria with inappropriately low serum concentration of parathyroid hormone (PTH). We report on a young boy who presented hypocalcemia with hypercalciuria, hyperphosphatemia and low serum concentration of PTH.Materials and MethodsGenomic DNA was extracted from peripheral venous blood of a pediatric patient with ADH. Molecular screening of the CASR coding sequence was performed by sequence analysis with an ABI PRISM® 3100 Avant Genetic Analyzer. Site-directed mutagenesis was performed directly on CASR wild type cDNA, cloned in pCR3.1 plasmid to obtain CASR mutant plasmid. Functional properties of mutant receptor were studied in transiently transfected HEK-293 cells with the wild-type or mutated CASR. The ERK phosphorylation levels were measured by western blot analysis.ResultsWe detected a novel heterozygous deletion of a cytosine (c.2682delC) causing a frameshift and a premature stop codon resulting in a truncation of the CaSR's C-tail. HEK-293 cells transfection with CASR Mutant increased significantly (P = 0.02) p-ERK levels by 3.8 fold versus CASR wild type.ConclusionWe identified a novel CASR gene mutation in a young boy with ADH. Although the mutation leads to a truncation of the protein, it leads to a constitutive gain-of-function of the receptor. This finding is in line with the clinical phenotype observed in our patient.

Purpose of review The calcium-sensing receptor gene (CaSR, chr. 3q13.3-21) is a candidate to explain nephrolithiasis. This review analyzes the potential role of CaSR in lithogenesis according to findings of functional and genetic studies. Recent findings CaSR is a cation receptor located in the tubular cell plasma membrane. Its activation decreases calcium reabsorption in the ascending limb and distal convoluted tubule, but increases phosphate reabsorption in proximal tubules and decreases water and proton reabsorption in collecting ducts. Its effects in proximal tubules and collecting ducts can limit the calcium phosphate precipitation risk induced by the increase in calcium excretion. The nonconservative CaSR gene Arg990Gly polymorphism was associated with nephrolithiasis and hypercalciuria in different populations. Arg990Gly is located on exon 7 and produces a gain of the CaSR function. rs7652589 and rs1501899 were also associated with nephrolithiasis in patients with normal citrate excretion. These polymorphisms are located in the CaSR gene regulatory region and may modify CaSR gene promoter activity. Summary The activating Arg990Gly polymorphism may predispose to nephrolithiasis by increasing calcium excretion. Polymorphisms at the regulatory region may predispose to nephrolithiasis by changing tubular expression of the CaSR. CaSR genotype may be a marker to identify patients prone to develop calcium nephrolithiasis.

An association between the R990G polymorphism of the CaSR gene, coding for calcium-sensing receptor, and primary hypercalciuria was found in kidney stone formers. To confirm this relationship, we investigated hypercalciuric women without stones and studied the effect of CaSR gene in human embryonic kidney cells (HEK-293). We genotyped for CaSR A986S, R990G, and Q1011E polymorphisms, 119 normocalciuric and 124 hypercalciuric women with negative history of kidney stones. Homozygous (n=2) or heterozygous (n=21) women for the 990G allele considered as one group had an increased risk to be hypercalciuric (odds ratio=5.2; P=0.001) and higher calcium excretion (P=0.005) in comparison with homozygous women for the 990R allele (n=220). HEK-293 cells were transfected with the variant allele at the three CaSR gene polymorphisms and with the most common allele with no variants. The transient increment of intracellular calcium caused by the stepwise increase of extracellular calcium was evaluated in stable transfected cells loaded with fura-2 AM. The extracellular calcium concentration producing the half-maximal intracellular calcium response was lower in HEK-293 cells transfected with the 990G allele than in those transfected with the wild-type allele (P=0.0001). Our findings indicate that R990G polymorphism results in a gain-of-function of the calcium-sensing receptor and increased susceptibility to primary hypercalciuria.