Connections between serum Trimethylamine N-Oxide (TMAO), a gut-derived metabolite, and vascular biomarkers evaluating arterial stiffness and subclinical atherosclerosis in children with obesity.

Introduction: Childhood obesity leads to early subclinical atherosclerosis and arterial stiffness. Studying biomarkers like trimethylamine N-oxide (TMAO), linked to cardio-metabolic disorders in adults, is crucial to prevent long-term cardiovascular issues. Methods: The study involved 70 children aged 4 to 18 (50 obese, 20 normal-weight). Clinical examination included BMI, waist measurements, puberty stage, the presence of acanthosis nigricans, and irregular menstrual cycles. Subclinical atherosclerosis was assessed by measuring the carotid intima-media thickness (CIMT), and the arterial stiffness was evaluated through surrogate markers like the pulse wave velocity (PWV), augmentation index (AIx), and peripheral and central blood pressures. The blood biomarkers included determining the values of TMAO, HOMA-IR, and other usual biomarkers investigating metabolism. Results: The study detected significantly elevated levels of TMAO in obese children compared to controls. TMAO presented positive correlations to BMI, waist circumference and waist-to-height ratio and was also observed as an independent predictor of all three parameters. Significant correlations were observed between TMAO and vascular markers such as CIMT, PWV, and peripheral BP levels. TMAO independently predicts CIMT, PWV, peripheral BP, and central SBP levels, even after adding BMI, waist circumference, waist-to-height ratio, puberty development and age in the regression model. Obese children with high HOMA-IR presented a greater weight excess and significantly higher vascular markers, but TMAO levels did not differ significantly from the obese with HOMA-IR

Unveiling the Silent Danger of Childhood Obesity: Non-Invasive Biomarkers Such as Carotid Intima-Media Thickness, Arterial Stiffness Surrogate Markers, and Blood Pressure Are Useful in Detecting Early Vascular Alterations in Obese Children.

Obese children present a higher cardio-metabolic risk. Measuring vascular biomarkers that assess the evolution of arterial stiffness, subclinical atherosclerosis, and hypertension in such patients could be helpful in the long term. We studied 84 children, aged from 6 to 18 years: 50 obese subjects, versus 34 of normal weight. Clinical examination involved: BMI, waist circumference, waist-to-height ratio, and detection of the presence of acanthosis nigricans and irregular menstrual cycles (the latter in adolescent girls). The carotid intima-media thickness (CIMT) was measured with the Aixplorer MACH 30 echography device. The pulse wave velocity (PWV), augmentation index (AIx), and peripheral and central blood pressures (i.e., SBP, DBP, cSBP, cDBP, and cPP) were acquired through a Mobil-O-Graph device. Obese subjects underwent body composition analysis with a Tanita BC-418. Blood tests were: HOMA-IR, lipid panel, uric acid, and 25-OH vitamin D. All vascular biomarkers presented increased values in obese subjects versus controls. The following cut-off values were significant in detecting obesity: for PWV > 4.6 m/s, cSBP > 106 mmHg for the <12-year-olds, PWV > 4.5 m/s and cSBP > 115 mmHg for the 12-15-year-olds, and PWV > 5 m/s, cSBP > 123 mmHg for the >15-year-olds. AIx is higher in obese children, regardless of their insulin resistance status. Waist circumference and waist-to-height ratio correlate to all vascular parameters. HOMA-IR is an independent predictor for all vascular parameters except CIMT. Cut-off values for PWV of >4.8 m/s, SBP > 125 mmHg, and a cSBP > 117 mmHg predicted the presence of acanthosis nigricans. Obese girls with irregular menses displayed significantly higher PWV, SBP, and DPB. Elevated levels of uric acid, LDL-c, non-LDL-c, triglycerides, and transaminases, and low levels of HDL-c and 25-OH vitamin D correlated with higher arterial stiffness and CIMT values. We conclude that CIMT and the markers of arterial stiffness are useful in the early detection of vascular damage in obese children.

A Comparative Analysis of Strain and 2D Shear Wave Elastography in the Diagnosis of Autoimmune Thyroiditis in Pediatric Patients.

This paper aims to assess the usefulness of shear-wave elastography (SWE) and strain elastography (SE) for identifying and monitoring thyroid gland changes in children diagnosed with chronic autoimmune thyroiditis (CAT). Our study included 77 children between the ages of six and eighteen. Of these, 45 were diagnosed with CAT, while 32 had no thyroid pathology. Following a clinical examination and laboratory tests, an ultrasound was carried out, and then a SE (using a Hitachi Preirus machine) and SWE (using an Aixplorer Mach 30, Supersonic imagine, France) were performed in the same session. The median thyroid elastic index (EI) in the CAT group was 13.8 (13.3-17) kPa compared to 10.1 (9.3-11.2) kPa in healthy children (p < 0.0001). We found a median strain ratio (SR) of 1.2 (1.2-1.3) for CAT compared to 0.7 (0.6-0.9) for healthy thyroid tissue (p < 0.0001). The optimal cut-off value for predicting the presence of CAT in children using SR was >1 (Se = 82.2%, Sp = 87.5%, PPV = 90.2%, and NPV = 77.8%, AUROC = 0.850), while using SWE, the optimal cut-off value for predicting the presence of CAT in children was >12 kPa (Se = 88.9%, Sp = 93.7%, PPV = 95.2%, and NPV = 85.5%, AUROC = 0.943). Both techniques are useful for measuring thyroid tissue elasticity, and their diagnostic accuracy and reliability are comparable.

Evaluating the Arterial Stiffness as a Useful Tool in the Management of Obese Children.

Childhood obesity speeds up the development of arterial stiffness and progressively increases the values of arterial pressure. The purpose of this study is to investigate the value of using pulse wave analysis (PWA) to measure arterial stiffness as a sign of vascular wall impairment in obese children. The research was focused on 60 subjects: 33 obese and 27 normal-weight. Ages ranged from 6 to 18 years old. PWA includes parameters such as pulse wave velocity (PWV), augmentation index (AIx), peripheral and central blood pressure (SBP, DBP, cSBP, cDBP), heart rate, and central pulse pressure (cPP). The device used was a Mobil-O-Graph. Blood parameters were taken from the subject's medical history, not older than 6 months. A high BMI and a large waist circumference are linked to a high PWV. The levels of LDL-c, triglycerides (TG), non-HDL-c, TG/HDL-c ratio, and total cholesterol-HDL-c ratio significantly correlate to PWV, SBP, and cSBP. Alanine aminotransferase is a reliable predictor of PWV, AIx, SBP, DBP, and cDBP, while aspartate aminotransferase is a significant predictor of AIx, mean arterial pressure (MAP), cSBP, and cPP. 25-OH-Vitamin D negatively correlates with PWV, SBP, and MAP and significantly predicts the MAP. Cortisol and TSH levels are not significant to arterial stiffness in obese children without specific comorbidities and neither is fasting glucose in obese children without impaired glucose tolerance. We conclude that PWA contributes valuable data regarding patients' vascular health and should be considered a reliable tool in the management of obese children.

Pregnancy Outcomes in SARS-CoV-2-Positive Patients: A 20-Month Retrospective Analysis of Delivery Cases.

Background and Objectives: The SARS-CoV-2 infection brings supplemental risks for pregnant women. Due to controversial hesitancy, their vaccination rate was lower in 2021 compared to the general population. In addition, access to maternal care was reduced during the pandemic. We conducted a retrospective cross-sectional analysis of the health records data over 20 months (1 April 2020 to 20 November 2021) aiming to explore the outcomes in SARS-CoV-2-positive cases referred for delivery to a tertiary public hospital in Western Romania. Materials and Methods: Women with SARS-CoV-2 infection diagnosed for the first time at the moment of birth who delivered singletons after 24 weeks of gestation, and had a clear immunization status were included in the analysis. Results: Out of the 97 patients included in the study, 35 (36%) had undergone ARN-based vaccination. Five cases of maternal death were recorded (all unvaccinated). Our retrospective exploratory analysis showed that the presence of COVID-19 symptoms in the SARS-CoV-2-positive patients made a significant impact on the delivery hospitalization, with a median hospital stay increase from 5 to 9 days (Mann-Whitney test, p = 0.014): longer hospitalization was recorded in the symptomatic cases irrespective of their vaccination status. No other adverse outcomes, such as gestational age at delivery, C-section rate, 5 min Apgar index, or birth weight were associated with the presence of symptoms. Conclusions: Our clinic maintained safe maternal care for the COVID-19 patients during the analyzed period. Vaccination of the expectant women was beneficial in SARS-CoV-2-positive patients by lowering the risk of COVID-19 symptoms, with subsequent implications on the newborns' health and maternal attachment.

Outcome of Newborns with Confirmed or Possible SARS-CoV-2 Vertical Infection-A Scoping Review.

Severe acute respiratory syndrome virus 2 (SARS-CoV-2), the virus that causes 2019 coronavirus disease (COVID-19), has been isolated from various tissues and body fluids, including the placenta, amniotic fluid, and umbilical cord of newborns. In the last few years, much scientific effort has been directed toward studying SARS-CoV-2, focusing on the different features of the virus, such as its structure and mechanisms of action. Moreover, much focus has been on developing accurate diagnostic tools and various drugs or vaccines to treat COVID-19. However, the available evidence is still scarce and consistent criteria should be used for diagnosing vertical transmission. Applying the PRISMA ScR guidelines, we conducted a scoping review with the primary objective of identifying the types, and examining the range, of available evidence of vertical transmission of SARS-CoV-2 from mother to newborn. We also aimed to clarify the key concepts and criteria for diagnosis of SARS-CoV-2 vertical infection in neonates and summarize the existing evidence and advance the awareness of SARS-CoV-2 vertical infection in pregnancy. Most studies we identified were case reports or case series (about 30% of poor quality and inconsistent reporting of the findings). Summarizing the existing classification criteria, we propose an algorithm for consistent diagnosis. Registration: INPLASY2022120093.

Multiparametric ultrasound-based assessment of overt hyperthyroid diffuse thyroid disease.

Introduction: Hyperthyroidismis a prevalent condition affecting global populations, with an overall prevalence of 1.2%. Our research aimed to establish a systematic diagnostic approach using multiparametric ultrasound (MPUS) to diagnose hyperthyroid diffuse thyroid disease (DTD). Methods: We conducted a retrospective study from June 2021 to June 2023 at a specialized endocrinology center in Timisoara, Romania, enrolling subjects presenting with clinical hyperthyroidism. Using the Mach 30 Aixplorer ultrasound equipment, evaluations were performed initially in B-mode US, followed by Color Doppler and Spectral Doppler measurements, and finally, 2D Shear wave elastography (SWE). Results: From the 218 patients analyzed, the diagnosis of DTD with hyperthyroidism was confirmed through biochemical assessment, subgrouping various pathologies such as subacute thyroiditis, Graves' disease, painless thyroiditis, Hashimoto's thyroiditis, iatrogenic, as well as healthy controls. In the first step, B-mode hypoechogenicity had an AUC of 0.951 for DTD detection. In the second step, the peak systolic velocity differentiated Graves' disease with a median of 42.4 cm/s and an AUC of 1. Lastly, the third step consisted of SWE evaluation, revealing a mean elasticity index in the SAT subgroup significantly higher from other subgroups (p<0.001) with an AUC of 1. Conclusion: Our study offers a step-by-step evaluation algorithm for DTD diagnosis, with a very good overall diagnostic performance (AUC of 0.946).

The Oscillometric Pulse Wave Analysis Is Useful in Evaluating the Arterial Stiffness of Obese Children with Relevant Cardiometabolic Risks.

Early detection of all complications of childhood obesity is imperative in order to minimize effects. Obesity causes vascular disruptions, including early increased arterial stiffness and high blood pressure. This study's aim is to assess the reliability of pulse wave analysis (PWA) in obese children and how additional risk factors influence the evaluated parameters. We analyzed 55 children aged 6-18 years old by measuring their pulse wave velocity (PWV), augmentation index (AIx), peripheral blood pressure (SBP, DBP), heart rate, central blood pressure (cSBP, cDBP) and central pulse pressure (cPP). We used the oscillometric IEM Mobil-O-Graph and performed a single-point brachial measurement. The subjects were divided into two groups: obese (n = 30) and normal-weight (n = 25) and were clinically and anamnestically assessed. BMI and waist circumference are significantly correlated to higher values for PWV, SBP, DBP, cSBP, and cDBP. Weight significantly predicts PWV, SBP, DBP and cPP. The risk factors that significantly influence the PWA and BP values are: a cardiometabolically risky pregnancy (higher PWV, AIx, SBP), active and passive smoking (higher PWV, SBP, cSBP, cDBP), sleep deprivation (higher PWV, SBP, cSBP) and sedentariness (higher PWV, AIx, peripheral and central BP). We conclude that obese children with specific additional cardiometabolic risk factors present increased arterial stiffness and higher blood pressure values.

Effects of Burosumab Treatment on Two Siblings with X-Linked Hypophosphatemia. Case Report and Literature Review.

X-linked hypophosphatemia (XLH) or vitamin D-resistant rickets (MIM#307800), is a monogenic disorder with X-linked inheritance. It is caused by mutations present in the Phosphate Regulating Endopeptidase Homolog X-Linked (PHEX) gene responsible for the degradation of the bone-derived hormone fibroblast growth factor 23 (FGF23) into inactive fragments, but the entire mechanism is currently unclear. The inactivation of the gene prevents the degradation of FGF23, causing increased levels of FGF23, which leads to decreased tubular reabsorbtion of phosphorus. Clinical aspects are growth delay, limb deformities, bone pain, osteomalacia, dental anomalies, and enthesopathy. Laboratory evaluation shows hypophosphatemia, elevated alkaline phosphatase (ALP), and normal serum calcium levels, whereas parathormone (PTH) may be normal or increased and FGF23 greatly increased. Conventional treatment consists of administration of oral phosphate and calcitriol. Treatment with Burosumab, a monoclonal antibody that binds to FGF23, reducing its activity, was approved in 2018. Methods. We describe a case of two siblings, a girl and a boy, diagnosed with XLH, monitored by the Genetic Department of the County Emergency Clinical Hospital since 2019. The clinical picture is suggestive for XLH, both siblings exhibiting short stature, lower limb curvature, bone pain, marked walking weakness, and fatigue. Radiological aspects showed marked deformity of the lower limbs: genu varum in the girl, genu varum and valgum in the boy. Laboratory investigations showed hypophosphathemia, hyperphosphaturia, elevated ALP, normal PTH, and highly increased FGF23 in both. DNA analysis performed on the two siblings revealed a nonsense mutation in exone 5 of the PHEX gene: NM_000444.6(PHEX):c.565C > T (p.Gln189Ter). Results. At the age of 13½ on 7 June 2021, the two children started treatment with Burosumab in therapeutic doses and were monitored clinically and biochemically at regular intervals according to the protocol established by the Endocrinology Commission of the Romanian Health Ministry. Conclusions. The first results of the Burosumab treatment in the two siblings are extremely encouraging and suggest a favorable long-term evolution under this treatment.

Postmortem MicroRNA Signatures as Predictive Markers in SARS-CoV-2 Infection.

BACKGROUND: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease that emerged in December 2019 from Wuhan, China, has led to a worldwide outbreak that has resulted in 234,809,103 confirmed cases and caused more than 4,800,375 deaths worldwide. MicroRNAs could be involved in the SARS-CoV-2 infection, but not many studies have been performed to explore this in postmortem cases. The purpose of our study is to evaluate the postmortem expression of microRNA-6501-5p, microRNA-5695, and microRNA-29b-3p from bronchial secretions in positive and negative SARS-CoV-2 deaths and to evaluate their usefulness as predictive biomarkers in the evolution of SARS-CoV-2 infection. METHODS: During the autopsy procedure on 61 "suspected" deaths at the Institute of Forensic Medicine in Timisoara, Romania, bronchial secretions were collected to detect SARS-CoV-2 infection postmortem. After the RT-PCR analysis, 44 SARS-CoV-2 cases were detected positive, while 17 cases were SARS-CoV-2 negative, which were considered as controls. RESULTS: From the panel of microRNAs, microRNA-6501-5p, microRNA-5695, and microRNA-29b-3p were upregulated in SARS-CoV-2 cases and down-regulated in non-SARS-CoV-2 cases. CONCLUSIONS: We concluded that using a panel of microRNAs as biomarkers in SARS-CoV-2 infection could aid in an early evaluation of the evolution of SARS-CoV-2-infected patients.

Methotrexate and Cetuximab-Biological Impact on Non-Tumorigenic Models: In Vitro and In Ovo Assessments.

Background Objectives: The neoplastic process remains a major health problem facing humanity. Although there are currently different therapeutic options, they raise a multitude of shortcomings related to the toxic effects associated with their administration. Methotrexate (Met) and Cetuximab (Cet) are two basic chemotherapeutics used in cancer practice, but notwithstanding despite many years of use, the mechanisms by which the multitude of side-effects occur are not yet fully understood. Thus, the present study focused on the in vitro and in ovo evaluation of the associated toxic mechanisms on keratinocytes, keys cells in the wound healing process. Materials and Methods: The two chemotherapeutics were tested in eight different concentrations to evaluate keratinocytes viability, the anti-migratory effect, and the influence on the expression of markers involved in the production of cell apoptosis. In addition, the potential irritating effect on the vascular plexus were highlighted by applying the in ovo method, chick chorioallantoic membrane (HET-CAM). Results: The results revealed that Met induced decreased cell viability as well as increased expression of pro-apoptotic genes. In the vascular plexus of the chorioallantoic membrane, Met caused vascular irritation accompanied by capillary hemorrhage and vascular stasis. Conclusions: Summarizing, Cet presents a safer toxicological profile, compared to Met, based on the results obtained from both in vitro (cell viability, wound healing, RT-PCR assays), and in ovo (HET-CAM assay) techniques.

Shear-Wave Elastography-Diagnostic Value in Children with Chronic Autoimmune Thyroiditis.

Chronic autoimmune thyroiditis (CAT) is the most common thyroid disorder in the pediatric population. Ultrasound evaluation may suggest the diagnosis. Additionally, shear-wave elastography (SWE) proved to be a valuable additional diagnosis tool in adults with CAT by assessing thyroid stiffness (TS). This study aims to assess its use also in detecting children with CAT. The study group consisted of 50 children with confirmed diagnosis of CAT, who were compared to the control group, consisting of 50 children with no thyroid pathology and with an adult group of 50 subjects with CAT. The evaluation included, besides bioimmunochemical evaluation, also thyroid ultrasound evaluation and elastography measurements in the same session (Aixplorer Mach 30, Supersonic imagine, France). The mean TS values were significantly lower for children in the CAT group compared to adults with CAT (15.51 ± 4.76 kPa vs. 20.96 ± 6.31 kPa; p < 0.0001) and higher compared to the healthy aged matched controls (15.51 ± 4.76 kPa vs. 10.41 ± 2.01 kPa; p < 0.0001). SWE elastography definitely seems a promising technique in the evaluation of children with autoimmune thyroid pathology.

CHDH-PNPLA3 Gene-Gene Interactions Predict Insulin Resistance in Children with Obesity.

INTRODUCTION: Insulin resistance plays a major role in metabolic syndrome and is recognized as the most common risk factor for non-alcoholic fatty liver disease (NAFLD). Identifying predictors for insulin resistance could optimize screening and prevention. PURPOSE: To evaluate the contribution of multiple single nucleotide polymorphisms across genes related to NAFLD and choline metabolism, in predicting insulin resistance in children with obesity. METHODS: One hundred fifty-three children with obesity (73 girls), aged 7-18 years, were evaluated within the NutriGen Study (ClinicalTrials.gov-NCT02837367). Insulin resistance was defined by Homeostatic Model Assessment for insulin-resistance cut-offs that accommodated pubertal and gender differences. Anthropometric, metabolic, intake-related variables, and 55 single nucleotide polymorphisms related to NAFLD and choline metabolism were evaluated. Gene-gene interaction effects were assessed using Multiple Data Reduction Software. RESULTS: Sixty percent (93/153) of participants showed insulin resistance (58.7% of boys, 63% of girls). Children with insulin resistance presented significantly higher values for standardized body mass index, triglycerides, transaminases and plasma choline when compared to those without insulin resistance. Out of 52 single nucleotide polymorphisms analysed, the interaction between genotypes CHDH(rs12676) and PNPLA3(rs738409) predicted insulin resistance. The model presented a 6/10 cross-validation consistency and 0.58 testing accuracy. Plasma choline levels and alanine aminotransferase modulated the gene interaction effect, significantly improving the model. CONCLUSION: The interaction between genotypes in CHDH and PNPLA3 genes, modulated by choline and alanine aminotransferase levels, predicted insulin-resistance status in children with obesity. If replicated in larger cohorts, these findings could help identify metabolic risk in children with obesity.

The Value of Strain Elastography in Predicting Autoimmune Thyroiditis.

Chronic autoimmune thyroiditis (CAT) defines a diffuse intrathyroidal lymphocytic infiltration associating a destructive process of the thyroid follicles, most commonly in evolution developing hypothyroidism. Typical ultrasound changes may suggest the presence of the disease. This study aims to evaluate the performance of strain elastography in detecting autoimmune thyroiditis as an additional tool to the conventional ultrasound examination. A total of 250 patients were enrolled in the study; 180 had biochemical confirmation of CAT, the other 70 healthy subjects represented the control group. All patients were examined clinically and by means of conventional thyroid ultrasound (US) and real-time elastography using a Hitachi Preirus machine (5-15 MHz linear probe). Five valid measurements for the parenchyma/muscle strain ratios (SR) were taken for each subject, considering the mean value for analysis. A mean SR value above 1.64 was found to predict the presence of CAT with sensitivity (Sen) 69%, specificity p92%, positive predictive value (PPV) 95.4%, negative predictive value (NPV) 54% and area under receiver operating characteristic (AUROC) 0.87. Moreover, when comparing the mean values for SR, significantly higher values were found in CAT patients compared with the controls (2.81 ± 2.11 vs. 1.03 ± 0.51; p < 0.0001). Of the 180 CAT subjects, 92 were on thyroid hormone replacement therapy; significantly higher values were detected for patients under therapy compared with asymptomatic cases (3.45 ± 2.53 vs. 2.15 ± 1.27, p < 0.0001). A cut-off value of 2.94 was established for identifying CAT patients who needed hormonal treatment (Sen 52.3%, Sp 83.7%, PPV 75.4%, NPV 64.7% and AUROC 0.66). No correlation was found between stiffness and antibody titers nor for functional status. Elastography does add valuable information to the US evaluation of cases with autoimmune thyroiditis.

Single Nucleotide Polymorphisms in PEMT and MTHFR Genes are Associated with Omega 3 and 6 Fatty Acid Levels in the Red Blood Cells of Children with Obesity.

Polyunsaturated fatty acids (PUFAs) play important roles in health and disease. PUFA levels are influenced by nutrition and genetic factors. The relationship between PUFA composition in red blood cells (RBCs) and genetic variations involved in PUFA metabolism has not been investigated in children with obesity. This study evaluated the association between several genetic variations and PUFA levels in RBCs in children with obesity. One hundred ninety-six children with obesity (101 females, 95 males) were evaluated using anthropometric measurements, dietary intakes, plasma and RBC PUFA quantification, blood biochemistry, and 55 single nucleotide polymorphisms within 14 genes. phosphatidylethanolamine N-methyltransferase (PEMT) rs1109859 and methylenetetrahydrofolate reductase gene (MTHFR) rs4846052 genotypes were associated with PUFA levels in RBCs. PUFA intake did not influence the RBC eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels. Higher RBC DHA and EPA levels were observed for PEMT rs1109859 GG and GA genotypes versus the AA genotype. Higher levels of RBC DHA, EPA, arachidonic acid (ARA), and linoleic acid (LA) and were observed for MTHFR rs4846052 TT genotype versus TC and CC genotypes. Genetic variations in PEMT rs1109859 and MTHFR rs4846052 were associated with different PUFA levels in RBC membranes and are estimators for PUFA species in RBCs. Further research is needed to establish whether these genotype-specific alterations are specific to overweight children.

A randomized controlled trial for overweight and obesity in preschoolers: the More and Less Europe study - an intervention within the STOP project.

BACKGROUND: Childhood overweight and obesity is a serious public health issue with an increase being observed in preschool-aged children. Treating childhood obesity is difficult and few countries use standardized treatments. Therefore, there is a need to find effective approaches that are feasible for both health care providers and families. Thus, the overall aim of this study is to assess the acceptance and effectiveness of a parent support program (the More and Less, ML) for the management of overweight and obesity followed by a mobile health (mHealth) program (the MINISTOP application) in a socially diverse population of families. METHODS/DESIGN: A two-arm, parallel design randomized controlled trial in 300 2-to 6-year-old children with overweight and obesity from Romania, Spain and Sweden (n = 100 from each). Following baseline assessments children are randomized into the intervention or control group in a 1:1 ratio. The intervention, the ML program, consists of 10-weekly group sessions which focus on evidence-based parenting practices, followed by the previously validated MINISTOP application for 6-months to support healthy eating and physical activity behaviors. The primary outcome is change in body mass index (BMI) z-score after 9-months and secondary outcomes include: waist circumference, eating behavior (Child Eating Behavior Questionnaire), parenting behavior (Comprehensive Feeding Practices Questionnaire), physical activity (ActiGraph wGT3x-BT), dietary patterns (based on metabolic markers from urine and 24 h dietary recalls), epigenetic and gut hormones (fasting blood samples), and the overall acceptance of the overweight and obesity management in young children (semi-structured interviews). Outcomes are measured at baseline and after: 10-weeks (only BMI z-score, waist circumference), 9-months (all outcomes), 15- and 21-months (all outcomes except physical activity, dietary patterns, epigenetics and gut hormones) post-baseline. DISCUSSION: This study will evaluate a parent support program for weight management in young children in three European countries. To boost the effect of the ML program the families will be supported by an app for 6-months. If the program is found to be effective, it has the potential to be implemented into routine care to reduce overweight and obesity in young children and the app could prove to be a viable option for sustained effects of the care provided. TRIAL REGISTRATION: ClinicalTrials.gov NCT03800823; 11 Jan 2019.

No clinical utility of common polymorphisms in IGF1, IRS1, GCKR, PPARG, GCK1 and KCTD1 genes previously associated with insulin resistance in overweight children from Romania and Moldova.

Background Previous genome-wide association studies (GWAS) identified IGF1, IRS1, GCKR, PPARG, GCK1 and KCTD1 as candidate genes for insulin resistance and type 2 diabetes (T2D). We investigated the associations of these previously reported common variants in these genes with insulin resistance in overweight children from Romania and Moldova. Methods Six single nucleotide polymorphisms (SNPs), IGF1 (rs35767), IRS1 (rs2943634), GCKR (rs780094), PPARG (rs1801282), GCK1 (rs1799884) and KCTD15 (rs29941), were genotyped in 100 overweight children along with clinical and metabolic parameters. Homeostatic model assessment of insulin resistance (HOMA-IR) above 3.4 (defining insulin resistance) was used as the outcome. Results Children differed in insulin resistance status despite having similar body mass index (BMI) standard deviation scores (SDS) (World Health Organization, [WHO] reference). The identified predictors for altered insulin metabolism were higher cholesterol levels, higher diastolic blood pressure and higher waist-to-hip-ratio (as a marker for increased abdominal fat). None of the SNPs showed significant association with increase in the risk for insulin resistance in children (p range=0.478-0.724; odds ratio [OR] range=1.924-4.842); however, the risk allele in GCKR (rs780094, p=0.06, OR=6.871) demonstrated near statistical significance. Conclusions The interrogated risk alleles did not show any significant association with insulin resistance in children in our cohort; however, the GCKR (rs780094) might be a viable candidate in larger cohorts. The lack of replication of the proposed association may point to differences in linkage disequilibrium or effect modifiers across studies.

Development and Validation of a LC⁻MS/MS-Based Assay for Quantification of Free and Total Omega 3 and 6 Fatty Acids from Human Plasma.

Few high-performance liquid chromatography⁻tandem mass spectrometry (LC-MS/MS) methods have been developed for the full quantitation of fatty acids from human plasma without derivatization. Therefore, we propose a method that requires fewer sample preparation steps, which can be used for the quantitation of several polyunsaturated fatty acids in human plasma. The method offers rapid, accurate, sensitive, and simultaneous quantification of omega 3 (α-linolenic, eicosapentaenoic, and docosahexaenoic acids) and omega 6 fatty acids (arachidonic and linoleic acids) using high-performance LC-MS/MS. The selected fatty acids were analysed in lipid extracts from both free and total forms. Chromatographic separation was achieved using a reversed phase C18 column with isocratic flow using ammonium acetate for improving negative electrospray ionization (ESI) response. Mass detection was performed in multiple reaction monitoring (MRM) mode, and deuterated internal standards were used for each target compound. The limits of quantification were situated in the low nanomolar range, excepting linoleic acid, for which the limit was in the high nanomolar range. The method was validated according to the U.S. Department of Health and Human Services guidelines, and offers a fast, sensitive, and reliable quantification of selected omega 3 and 6 fatty acids in human plasma.

Congenital anomalies of digits - a clinical-epidemiological study of 301 patients.

INTRODUCTION: Congenital anomalies of digits (CAD) can occur as isolated malformations, in combination with other malformation of the limbs, or as part of a genetic syndrome. The purpose of this work is to provide an overview of CAD, on morphological, genetic and epidemiological basis. PATIENTS AND METHODS: We conducted a retrospective analysis of a cohort of 301 patients with CAD. Following the Swanson classification, the list of anomalies under study included: adactyly and oligodactyly, syndactyly and symphalangism, polydactyly, macrodactyly, amniotic bands syndrome, and generalized skeletal anomalies. RESULTS: In Bihor County, Romania, the Department of Medical Genetics recorded 4916 patients with congenital anomalies (2.03% out of 241 601 live newborns) between 1984 and 2018. Of these, 301 (6.1%) patients had CAD. The prevalence of CAD was 1:800 living newborns. The most common CAD were polydactyly, followed by syndactyly, brachydactyly, adactyly and oligodactyly. Upper extremities were four times more frequently affected than lower extremities, while both upper and lower extremities were affected in a quarter of all cases. CAD were isolated in 64% of patients, while 14% were associated with other anomalies of the extremities and 22% were associated with recognized genetic syndromes. CONCLUSIONS: Our study, by its size and the long period of clinical observation, provides opportunities to generalize and compare our data with similar studies, offering the possibility for improved knowledge of the epidemiology of CAD and potential improvements in genetic counseling.

A novel method for measuring subcutaneous adipose tissue using ultrasound in children - interobserver consistency.

BACKGROUND: Currently, sufficiently accurate field methods for body composition assessment in children are missing. The ultrasound method for assessing adipose tissue thickness has been used extensively in sport medicine. However, there are no studies looking at the reliability of this method in non-athletic children. This paper aims to determine the inter-observer reliability in measuring the uncompressed subcutaneous adipose tissue thickness using ultrasound, in children. SUBJECTS, MATERIALS AND METHODS: Forty healthy children (20 males, 20 females), median age 11.85 years (5.3 to 18.1 years) were evaluated. Median body mass index standard deviation score (BMI SDS) was -0.13 (-3.9 to 4). Three observers used a Hosand BX 2000 Ultrasonic Adipometer to measure uncompressed subcutaneous adipose tissue thickness at three sites: triceps, subscapular, supraspinale. A single experienced observer used the three sites to also measure the compressed adipose thickness using a skinfold caliper. RESULTS: Individual observer deviations from the mean value of the three observers in adipometer measurement had a standard deviation of 1.74 mm, 92.8% were less than 3 mm. Analysis separated by individual anatomical sites showed high reliability values for triceps: linear regression R²=0.84, p=0.000; intraclass correlation coefficient (ICC)=0.92 and standard error of measurement (SEM)=0.63. The values at supraspinale site were R²=0.82, p=0.000, ICC=0.89 and SEM=1.17, while for subscapular the values were lower: R²=0.79, p=0.000, ICC=0.78 and SEM=1.02. The body fat percentage (BF%) calculated using skinfold measurements was highly correlated with the BF% calculated by the adipometer (R=0.92, R²=0.83, p=0.000). The Pearson's correlation between BMI SDS and BF% calculated from skinfold was R=0.52, R²=0.28, p=0.001, while for the adipometer it was R=0.53, R²=0.27, p=0.000. CONCLUSIONS: This novel ultrasound measurement technique can be used with good accuracy and reliability to measure uncompressed subcutaneous adipose tissue thickness in children, sustaining its application for research and clinical purposes, however larger studies are needed.