ICH3, a selective alpha7 nicotinic acetylcholine receptor agonist, modulates adipocyte inflammation associated with obesity.

PURPOSE: The alpha7 nicotinic acetylcholine receptor (α7nAChR), involved in the modulation of inflammation and insulin sensitivity, is downregulated in white adipose tissue (WAT) of obese patients. This study aims to test the ability of a selective synthetic α7nAChR agonist, the spirocyclic Δ2-isoxazoline derivative (R)-(-)-ICH3 (ICH3), to counteract acute inflammation and obesity-associated modifications in WAT. METHODS: We employed the LPS-septic shock murine model, human primary adipocytes and diet-induced obese (DIO) mice. Inflammatory factor expression was assessed by ELISA and quantitative real-time PCR. Flow cytometry was employed to define WAT inflammatory infiltrate. Insulin signaling was monitored by quantification of AKT phosphorylation. RESULTS: In the septic shock model, ICH3 revealed antipyretic action and reduced the surge of circulating cytokines. In vitro, ICH3 stimulation (10 µM) preserved viability of human adipocytes, decreased IL-6 mRNA (P < 0.05) and blunted LPS-induced peak of TNFα (P < 0.05) and IL-6 (P < 0.01). Chronic administration of ICH3 to DIO mice was associated with lower numbers of CD8+ T cells (P < 0.05) and to changed WAT expression of inflammatory factors (Hp, P < 0.05; CD301/MGL1, P < 0.01; Arg-1, P < 0.05). As compared to untreated, ICH3 DIO mice exhibited improved insulin signaling in the skeletal muscle (P < 0.01) mirrored by an improved response to glucose load (ipGTT: P < 0.05 at 120 min). CONCLUSIONS: We proved that ICH3 is an anti-inflammatory drug, able to reduce inflammatory cytokines in human adipocytes and to blunt the effects of obesity on WAT inflammatory profile, on glucose tolerance and on tissue insulin sensitivity.

Nordic walking increases circulating VEGF more than traditional walking training in postmenopause.

OBJECTIVES: Nordic walking (NW) is widely practiced by postmenopausal women. Its effects are peculiar owing to the involvement of more muscle groups than in traditional walking training (WT). Since mechanical load promotes secretion of vascular endothelial growth factor (VEGF) from both skeletal muscle and muscle endothelium, the aim of the study was to compare the effect of NW and WT on VEGF levels. METHOD: Thirty postmenopausal women were randomly assigned to NW or WT. Both groups trained 40-50 min/day, three times per week, at a mean intensity of 12 on a 15-category scale of the ratings of perceived exertion. Since VEGF is also released from adipocytes, anthropometric parameters were assessed. RESULTS: NW increased circulating VEGF more than WT (p = 0.041). Furthermore, both study groups exhibited an average decrease in weight (p = 0.023), body mass index (p = 0.024), hip circumference (p = 0.001), and arm fat index, although WT participants had higher values for this index at baseline (p < 0.001) and thus exhibited a greater net decrease compared with the NW participants (p < 0.011). CONCLUSIONS: These data imply that NW increases the level of circulating VEGF more than does traditional walking when the intensity of training is equivalent.

Adipose tissue microbiota in humans: an open issue.

BACKGROUND: A specific 'adipose tissue' microbiota has been recently identified in mice and hypothesized in humans. The purpose of this study was to verify the presence of microbiota of human whole adipose tissue and isolated adipocytes by combining culture-dependent and independent methods. METHODS: Standard microbiological cultural techniques and 16S ribosomal RNA (16S rRNA) gene sequencing (Illumina technology) on DNA and RNA were employed to study (a) whole abdominal subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) from 14 obese and five normal-weight subjects and (b) mature adipocytes isolated from SAT and VAT after collagenase digestion or mechanical separation. To optimize the 16S rRNA gene detection, we used different DNA extraction methods (lysis with proteinase K, proteinase K+lysozyme and microbeads) and amplification procedures (semi-quantitative standard PCR and real-time quantitative PCR). RESULTS: Microbiological cultures were negative in all analyzed samples. In enzymatically isolated adipocytes, 90% of the sequenced bacterial DNA belonged to Clostridium histolyticum, the bacterium from which the collagenase enzyme was isolated. Bacterial 16S rRNA gene was not detected from DNA and RNA of whole SAT and VAT, as well as of mechanically isolated mature adipocytes, even after blocking with a specific primer the nonspecific amplification of human mitochondrial 12S rRNA. CONCLUSIONS: Our results do not support the presence of a human adipose tissue microbiota. In addition, they emphasized the technical problems encountered when applying metagenomic studies to human tissues with very low or absent bacterial load.

Determination of vertical dimension in prosthodontic rehabilitation of a growing patient with severe oligodontia.

CASE REPORT: This report describes the oral rehabilitation of a young boy with severe oligodontia of primary dentition and complete anodontia of permanent dentition, treated by complete maxillary and partial mandibular removable dentures. Since the determination of a proper occlusal vertical dimension in paediatric patients is often difficult, treatment was achieved by means of a multidisciplinary approach involving conventional prosthodontic principles combined with cephalometric analysis and growth prediction. After a follow-up of two years a second lateral cephalogram was taken to evaluate the developmental changes in craniofacial morphology and the superimpositions method was performed, showing an improvement of both sagittal and vertical relationship between basal bones. In this case, the early prosthetic treatment resulted in enhanced aesthetics and masticatory function as well as skeletal growth changes.

Lifestyle and high density lipoprotein cholesterol in postmenopause.

OBJECTIVES: Menopause is characterized by hormonal and metabolic changes. These are linked to increased risk of cardiovascular disease, for which low blood plasma levels of high density lipoprotein (HDL) cholesterol are an independent risk factor. The present study investigated variables linked with basal plasma HDL cholesterol levels and the effects of aerobic training, on their variations, in 40 postmenopausal women. METHODS: We assessed body composition, dietary habits and maximal aerobic capacity of participants. Characteristics of daily physical activity and plasma lipoproteins were measured. The women walked on 4 days/week, for 14 weeks, at moderate intensity, and they were grouped according to the resulting tertiles of basal plasma HDL cholesterol levels. RESULTS: Logistic regression analysis showed that waist-to-hip ratio and number of daily bouts of moderate-intensity physical activity, held for at least 10 consecutive minutes (B10m/day), are predictive variables of basal plasma HDL cholesterol levels. After the training period, the first and second tertiles increased plasma HDL cholesterol levels, while the third tertile decreased plasma HDL cholesterol levels. The tertiles showed different remodelling of spontaneous physical activity: the third tertile reduced B10m/day, while the others did not. CONCLUSIONS: This study provides knowledge about the relationships of plasma HDL cholesterol levels with characteristics of physical activity. Furthermore, it shows that physical exercise engagement can result in negative compensation of spontaneous physical activity that could counteract or reduce the positive effects of the aerobic training on plasma HDL cholesterol levels.

A promising eco-sustainable wound dressing based on cellulose extracted from Spartium junceum L. and impregnated with Glycyrrhiza glabra L extract: Design, production and biological properties.

Glycyrrhiza glabra extract is widely known for its antioxidant and anti-inflammatory properties and can improve the wound healing process. The aim of this work was to shorten the time of the healing process by using an eco-sustainable wound dressing based on Spanish broom flexible cellulosic fabric by impregnation with G. glabra extract-loaded ethosomes. Chemical analysis of G. glabra extract was performed by LC-DAD-MS/MS and its encapsulation into ethosomes was obtained using the ethanol injection method. Lipid vesicles were characterized in terms of size, polydispersity index, entrapment efficiency, zeta potential, and stability. In vitro release studies, biocompatibility, and scratch test on 3T3 fibroblasts were performed. Moreover, the structure of Spanish broom dressing and its ability to absorb wound exudate was characterized by Synchrotron X-ray phase contrast microtomography (SR-PCmicroCT). Ethosomes showed a good entrapment efficiency, nanometric size, good stability over time and a slow release of polyphenols compared to the free extract, and were not cytotoxic. Lastly, the results revealed that Spanish broom wound dressing loaded with G. glabra ethosomes is able to accelerate wound closure by reducing wound healing time. To sum up, Spanish broom wound dressing could be a potential new green tool for biomedical applications.

Permanence of molecular features of obesity in subcutaneous adipose tissue of ex-obese subjects.

OBJECTIVE: Bariatric surgery represents a powerful tool for morbid obesity treatment. However, after stabilization of weight loss that follows surgical interventions, ex-obese patients face the problem of residual tissues removal. Actually, it is unknown whether the characteristics of this residual subcutaneous adipose tissue (SAT) are 'restored' with regard to molecular and morphological features. DESIGN: To clarify this issue, we compared the SAT gene expression profile of ex-obese patients (ExOB-SAT, mean body mass index (BMI): 27.2±1.3 kg m(-2)) with that of lean (normal weight, NW-SAT, mean BMI: 22.6±1.1 kg m(-2)), overweight (OW-SAT, BMI: 27.65±0.2 kg m(-2)) and obese patients, according to BMI classes (OB1-SAT: 30 > or = BMI < or = 34.9, OB2-SAT: 35 > or = BMI < or = 39.9, OB3-SAT: BMI > or = 40). SUBJECTS AND METHODS: A total of 58 samples of SAT were collected during surgical interventions. Gene expression levels were assessed by microarrays and significant genes were validated by RT-qPCR. Adipocyte hypertrophy, inflammatory infiltration and fibrosis were assessed by morphological techniques. RESULTS: Global gene expression in ExOB-SAT was closely related to gene expression of OB3-SAT by hierarchical clustering procedures, in spite of different BMI. Metallothioneins (MT1A and MT2A) were the key over-expressed genes in both groups. At morphologic level, adipocyte hypertrophy and inflammatory infiltration improved after weight loss in ExOB-SAT, despite a persistence of fibrosis. CONCLUSIONS: Taken together, these results demonstrate that SAT gene expression is not fully restored, even after an extensive and stable weight loss. The persistence of 'obesity molecular features' in ExOB-SAT suggests that the molecular signature of adipose tissue is not solely dependent on weight loss and may need longer time period to completely disappear.

Characteristics of spontaneous physical activity and executive functions in postmenopause.

AIM: Executive functions are susceptible to age-related changes, and menopause has also been shown to be linked with their decline. The aim of the present study was to investigate the variables related to executive functions in postmenopausal women not involved in controlled dietary and physical exercise programs and without hormone-replacement therapy. METHODS: Fifty-seven women (58.39 ±4.16 yr) were investigated for their medical history, blood lipids, insulin resistance (HOMA-IR), body composition, blood pressure, executive functions (Digit Span and Trail Making tests), maximal aerobic capacity, dietary habits, and spontaneous physical activity. RESULTS: Cluster analysis performed on the basal data of the Digit Span scores and Trail Making tests identified two subgroups: Group A (N.=24) that showed better executive functions than Group B (N.=33). Although these subgroups differed across several variables of body composition, plasma lipids, dietary habits, spontaneous physical activity, aerobic fitness, and insulin resistance, logistic regression models showed B10m/die as the only predictive variable for subgroup membership. CONCLUSION: In the absence of a physical exercise program, the knowledge of how spontaneous physical activity is organized (B10m/die) is important for executive functions of post-menopausal women. It is the specific moderate to intense physical activity characteristic of B10m/die, rather than generic physical activity, that seems to determine the maintenance or attainment of cognitive health through body movement.

Temporomandibular joint dysfunction in Moebius syndrome.

AIM: Moebius syndrome is a rare condition characterised by bilateral facial and abducens nerve paralysis. In the present study, it was investigated the effect of the long term facial muscles hypoactivity on temporomandibular joint movements development. METHODS: Accordingly with Terzis classification (Terzis, 2003), a wide sample of A type and B type Moebius patients was investigated for mandibular range of movements. Moebius patients were compared with a sample of healthy subjects. RESULTS: Both type A and type B Moebius patients develop a severe articular movement reduction, especially during mouth opening. A highly significant difference was found between Moebius patients and the healthy group. No difference was found between A and B Moebius sub-samples. CONCLUSION: The authors stress the importance of an early adequate rehabilitation of Moebius patients, in order to avoid the limitation of TMJ movement range.

The nicotinic acetylcholine receptor α7 in subcutaneous mature adipocytes: downregulation in human obesity and modulation by diet-induced weight loss.

BACKGROUND: It is known that cholinergic anti-inflammatory reflex regulates inflammation in peripheral tissues. Nicotinic acetylcholine receptors (nAChRs) are mediators of this anti-inflammatory pathway and also non-neuronal cells express functional nAChrs. A role for α7-subtype acetylcholine cholinergic receptor (α7nAChR) in insulin sensitivity improvement has already been shown in rodents both in vivo and in vitro. However, no data are available on α7nAChR expression in human adipocytes. OBJECTIVE: To investigate the expression and protein content of α7nAChR in human subcutaneous adipose tissue (SAT) and in isolated mature adipocytes. DESIGN: A total of 39 SAT biopsy specimens obtained from obese and normal-weight subjects were used to assess α7nAChR messenger RNA levels and to stimulate α7nAChR with a specific agonist and antagonist in vitro. Additional SATs from eight non-diabetic obese subjects were also studied, before and after a 3-month lifestyle intervention. RESULTS: α7nAChR expression was significantly lower in the SAT of obese subjects compared with that of normal-weight subjects. In mature adipocytes isolated from morbidly obese subjects (body mass index > 40 kg m(-2)), α7nAChR expression was 75% lower compared with adipocytes from normal-weight subjects. In adipocytes of obese subjects, α7nAChR was downregulated also at protein level. In eight non-diabetic obese subjects, a lifestyle intervention (3 months of diet and physical activity) induced a significant weight loss and an increase in α7nAChR SAT expression. In vitro stimulation of adipocytes with the specific α7nAChR agonist PNU282987 induced a significant anti-inflammatory effect. Furthermore, a similar downregulation of the inflammatory profile, associated with a significant increase in α7nAChR protein level, was observed after genistein stimulation. CONCLUSIONS: These results provide evidence that α7nAChR expression levels are significantly decreased in obese subjects, and that this receptor modulates inflammatory gene expression in human adipocytes. The upregulation of α7nAChR by genistein stimulation opens new insights for the management of low-grade inflammation linked to human obesity.

[Determinants of short-term memory: is there a difference between young adults and postmenopausal women?]. / Fattori determinanti la memoria a breve termine: esiste una differenza tra giovani donne e donne in postmenopausa?

AIM: The literature indicates that several variables are influencing cognitive health. The aim of the study was to investigate the main determinants of the short-term memory among anthropometric, dietary and performance variables in a sample of healthy women. The role played by the age was also investigated. METHODS: Forty-five healthy overweight women were recruited through general physicians: 23 were young adults (24.63±4.17 years) and 22 were postmenopausal (53.30±2.95 years). Overweight condition was assessed according to the age-adjusted reference values. Participants were analyzed for Digit Span, blood pressure, body composition, aerobic fitness and dietary habits. RESULTS: Young adults and postmenopausal women did not differ either in Digit Span or in dietary habits. In postmenopausal women Digit Span was positively correlated with body weight, body mass index, body fat, waist circumference and daily intake of vitamin D. Linear regression model indicated vitamin D as the only significant predictor variable of Digit Span. In young adults Digit Span had no correlations with the others investigated variables. CONCLUSION: In postmenopause, vitamin D daily intake is important not only for skeletal, but also for cognitive health. Even though young adults and post-menopausal women did not differ for health status, short-term memory in young adulthood seems to be differently linked with the investigated variables than during post-menopause.

Relationships between physical exercise practice, dietary behaviour and body composition in female university students.

AIM: The relationship between female body composition, total energy expenditure and physical exercise energy expenditure is not very strong because women can compensate by increasing their energy intake. The aim of this study was to investigate the relationship between total minutes of exercise per week, dietary habits and body composition in female university students using physical activity and the habit to cook as covariates. METHODS: Fifty-one female university students were investigated for body composition, dietary habits, aerobic fitness, physical activity and physical exercise practice. Participants were grouped in three sub-samples (1, 2 and 3) according to the 33rd and 66th percentiles of weekly minutes (min/wk) of training. RESULTS: Group 1 had 0.00 min/wk, group 2 had 95±35.59 min/wk and group 3 had 231.66±85.97 min/wk of training. Sub-samples did not differ for IPAQ parameters while differed for aerobic fitness (P=0.001). Group 3 had a higher energy intake (EI) (P=0.008), a higher intake of lipids (P=0.017), saturated (P=0.042) and monounsaturated fat (P=0.024) and a lower intake of carbohydrates (P=0.007). Group 3 maintained the higher EI and the worse composition of lipid intake considering the habit to cook as covariate. Group 3 also had higher muscle mass. CONCLUSION: In order to positively affect body composition, in the sedentary women it is enough to control the energy balance, whereas in those that trained regularly it is necessary to control both energy balance and composition of daily meals.

Effects of the time of day of walking on dietary behaviour, body composition and aerobic fitness in post-menopausal women.

AIM: Post-menopause is linked to an increased risk of death from cardiovascular disease. Physical exercise and healthy dietary habits are normally suggested to enhance health. The aim of this study was to verify whether the time of day of walking had different effects on both spontaneous dietary intake and body composition modification in overweight and sedentary post-menopausal women. METHODS: Forty-two sedentary post-menopausal women (53.46+/-3.32 yrs) were recruited. Thirty-three completed the study: 29 were suitable for statistical analysis. Of those, 14 walked in the morning (MG) and 15 in the early evening (EG). Body composition, dietary habits and predicted VO2max were investigated. Food intake was analysed for energy, macronutrients and daily distribution. RESULTS: The Mann-Whitney test showed that according to the time of day of walking there were different fat mass (FM) reductions and dietary behaviour responses. EG reduced FM greater than MG and showed a major increase in morning energy intake (EI). Sub-samples did not differ in total EI, daily macronutrient portioning and daily meals variations. The variation of FM was correlated with that of proteins (r=-0.352), morning EI (r=-0.367) and aerobic performance (r=0.369). Both MG and EG improved their aerobic performance. CONCLUSION: The positive effects of walking on health could be optimised by its evening execution because it could also be linked to spontaneous dietary habit modification.

Wild rats as urban detectives for latent sources of asbestos contamination.

Based on a large body of evidence asbestos minerals have been classified as carcinogens. Despite the Italian ban on asbestos in 1992 and the subsequent remediation activities, latent sources of contamination may still represent a hazard where asbestos were particularly used. Using wild rats as sentinel animals, this study aimed at uncovering sites with the greatest potential for non-occupational exposure to asbestos in the city of Casale Monferrato (Piedmont Region, Italy), where the largest Italian manufacturing plant of asbestos-cement had been active. During the study period (2013-2015) a total of 40 wild rats were captured from 16 sampling capture points. The lungs of wild rats have been investigated by using scanning electron microscopy (SEM) with energy dispersive spectroscopy (EDS). The SEM-EDS detected the presence of asbestos fibers (tremolite/actinolite, amosite, and chrysotile) in rats' lungs from 11 sampling points. The hypothetical rats' home-range and the observed site-specific concentration of asbestos fibers per gram of dry lung tissue were used to identify areas to be targeted by additional search of latent sources of asbestos. In conclusion, our results showed that the use of wild rats as sentinel animals may effectively integrate the strategies currently in use to reduce the exposure to asbestos.

Are physiological characteristics of Caribbean dance useful for health?

AIM: Although the current literature underlines the main role of physical inactivity in the development of chronic diseases and premature death, 65% of adults do not reach the minimum movement levels required to maintain and improve health. The aim of this study was to investigate whether the metabolic and cardiocirculatory characteristics of a single lesson of Caribbean dance fit with international recommendations to improve health through movement. METHODS: Energy expenditure, exercise intensity, mean heart rate and blood pressure response to a 90-minute lesson were analysed in 24 beginner and 24 experienced dancers (mean age 33.93+/-9.98 years). RESULTS: MANCOVA was used to analyse energy expenditure and exercise intensity of our sample, stratified for gender and experience. Body weight was inserted as a covariate. Experienced had a major total energy expenditure (372.75+/- 75.32 vs 297.33+/-87.54 kcal; P<0.001), minutes of exercise intensity >6 metabolic equivalents (METs) (8.04+/-10.65 vs 1.47+/-2.16; P<0.001), mean METs (3.81+/-0.32 vs 3.42+/-0.37; P<0.05) and mean heart rate (60.68+/-1.35 vs 53.24+/-3.91 %HRR; P<0.001) than beginner dancers. There were no statistical differences in blood pressure values. CONCLUSIONS: Caribbean dance fits with international guidelines to improve health and can aid the promotion and enhancement of health through its physiological characteristics, and may reduce drop-out due to a reduced motivation to move.

Midline mandibular osteotomy in an asymmetric patient.

This case report shows the possibility of the application of a mandibular osteotomy to resolve mandibular asymmetry with independent and discordant movements of both bony segments. The authors report the case of a 25-year-old woman referred for mandibular asymmetry, with a transverse excess of the right hemi mandible and vertical defect of the left one. The patient underwent a bilateral sagittal split osteotomy, midline osteotomy, and genioplasty, which corrected the mandibular asymmetry with contraction of the entire right hemi mandible. A slight left vertical increase was also obtained through the surgically created lateral open bite. In the follow-up assessment, the patient's face appeared symmetrical with normalization of the bizygomatic-bigonial relationships, and the facial shape corresponded to ideal anthropometric features. This technique resulted in resolution of mandibular asymmetry. In addition, mandibular osteotomy permits the esthetic management of the shape of the entire mandibular body in relation to the other third of the face.

Facial aesthetics and childhood.

AIM: The aim of the study is to evaluate attention and perception of parents about children's facial aesthetics, the paediatrician's role in early diagnosis of these problems and the areas of the face considered as the most important from an aesthetic point of view. METHODS: The study was carried out collecting 1214 questionnaires at paediatricians waiting room in the North of Italy. RESULTS: Most parents showed attention toward craniofacial deformities of their children and considered the paediatrician's role as very important in early diagnosis. Profile was considered more important than the front view and face's areas felt as the most important were denture, eyes and nose. CONCLUSION: Despite public opinion, adults are concerned not only about the general health and growth of their children, but also about their facial aesthetics. They, in fact, not only judge aesthetics of the whole face, but also discriminate different views and specific areas of the face. They require, for early diagnosis of dentofacial problems, skilled attention from the paediatrician.

Population structure of modern-day Italians reveals patterns of ancient and archaic ancestries in Southern Europe.

European populations display low genetic differentiation as the result of long-term blending of their ancient founding ancestries. However, it is unclear how the combination of ancient ancestries related to early foragers, Neolithic farmers, and Bronze Age nomadic pastoralists can explain the distribution of genetic variation across Europe. Populations in natural crossroads like the Italian peninsula are expected to recapitulate the continental diversity, but have been systematically understudied. Here, we characterize the ancestry profiles of Italian populations using a genome-wide dataset representative of modern and ancient samples from across Italy, Europe, and the rest of the world. Italian genomes capture several ancient signatures, including a non-steppe contribution derived ultimately from the Caucasus. Differences in ancestry composition, as the result of migration and admixture, have generated in Italy the largest degree of population structure detected so far in the continent, as well as shaping the amount of Neanderthal DNA in modern-day populations.

Multifactorial analysis of the stochastic epigenetic variability in cord blood confirmed an impact of common behavioral and environmental factors but not of in vitro conception.

Background: An increased incidence of imprint-associated disorders has been reported in babies born from assisted reproductive technology (ART). However, previous studies supporting an association between ART and an altered DNA methylation status of the conceived babies have been often conducted on a limited number of methylation sites and without correction for critical potential confounders. Moreover, all the previous studies focused on the identification of methylation changes shared among subjects while an evaluation of stochastic differences has never been conducted. This study aims to evaluate the effect of ART and other common behavioral or environmental factors associated with pregnancy on stochastic epigenetic variability using a multivariate approach. Results: DNA methylation levels of cord blood from 23 in vitro and 41 naturally conceived children were analyzed using the Infinium HumanMethylation450 BeadChips. After multiple testing correction, no statistically significant difference emerged in the number of cord blood stochastic epigenetic variations or in the methylation levels between in vitro- and in vivo-conceived babies. Conversely, four multiple factor analysis dimensions summarizing common phenotypic, behavioral, or environmental factors (cord blood cell composition, pre or post conception supplementation of folates, birth percentiles, gestational age, cesarean section, pre-gestational mother's weight, parents' BMI and obesity status, presence of adverse pregnancy outcomes, mother's smoking status, and season of birth) were significantly associated with stochastic epigenetic variability. The stochastic epigenetic variation analysis allowed the identification of a rare imprinting defect in the locus GNAS in one of the babies belonging to the control population, which would not have emerged using a classical case-control association analysis. Conclusions: We confirmed the effect of several common behavioral or environmental factors on the epigenome of newborns and described for the first time an epigenetic effect related to season of birth. Children born after ART did not appear to have an increased risk of genome-wide changes in DNA methylation either at specific loci or randomly scattered throughout the genome. The inability to identify differences between cases and controls suggests that the number of stochastic epigenetic variations potentially induced by ART was not greater than that naturally produced in response to maternal behavior or other common environmental factors.

Genome-wide DNA methylation analysis in blood cells from patients with Werner syndrome.

BACKGROUND: Werner syndrome is a progeroid disorder characterized by premature age-related phenotypes. Although it is well established that autosomal recessive mutations in the WRN gene is responsible for Werner syndrome, the molecular alterations that lead to disease phenotype remain still unidentified. RESULTS: To address whether epigenetic changes can be associated with Werner syndrome phenotype, we analysed genome-wide DNA methylation profile using the Infinium MethylationEPIC BeadChip in the whole blood from three patients affected by Werner syndrome compared with three age- and sex-matched healthy controls. Hypermethylated probes were enriched in glycosphingolipid biosynthesis, FoxO signalling and insulin signalling pathways, while hypomethylated probes were enriched in PI3K-Akt signalling and focal adhesion pathways. Twenty-two out of 47 of the differentially methylated genes belonging to the enriched pathways resulted differentially expressed in a publicly available dataset on Werner syndrome fibroblasts. Interestingly, differentially methylated regions identified CERS1 and CERS3, two members of the ceramide synthase family. Moreover, we found differentially methylated probes within ITGA9 and ADAM12 genes, whose methylation is altered in systemic sclerosis, and within the PRDM8 gene, whose methylation is affected in dyskeratosis congenita and Down syndrome. CONCLUSIONS: DNA methylation changes in the peripheral blood from Werner syndrome patients provide new insight in the pathogenesis of the disease, highlighting in some cases a functional correlation of gene expression and methylation status.