Integration of CTCF Loops, Methylome, and Transcriptome in Differentiating LUHMES as a Model for Imprinting Dynamics of the 15q11-q13 Locus in Human Neurons.

Human cell line models, including the neuronal precursor line LUHMES, are important for investigating developmental transcriptional dynamics within imprinted regions, particularly the 15q11-q13 Angelman (AS) and Prader-Willi (PWS) syndrome locus. AS results from loss of maternal UBE3A in neurons, where the paternal allele is silenced by a convergent antisense transcript UBE3A-ATS, a lncRNA that normally terminates at PWAR1 in non-neurons. qRTPCR analysis confirmed the exclusive and progressive increase in UBE3A-ATS in differentiating LUHMES neurons, validating their use for studying UBE3A silencing. Genome-wide transcriptome analyses revealed changes to 11,834 genes during neuronal differentiation, including the upregulation of most genes within the 15q11-q13 locus. To identify dynamic changes in chromatin loops linked to transcriptional activity, we performed a HiChIP validated by 4C, which identified two neuron-specific CTCF loops between MAGEL2-SNRPN and PWAR1-UBE3A. To determine if allele-specific differentially methylated regions (DMR) may be associated with CTCF loop anchors, whole genome long-read nanopore sequencing was performed. We identified a paternally hypomethylated DMR near the SNRPN upstream loop anchor exclusive to neurons and a paternally hypermethylated DMR near the PWAR1 CTCF anchor exclusive to undifferentiated cells, consistent with increases in neuronal transcription. Additionally, DMRs near CTCF loop anchors were observed in both cell types, indicative of allele-specific differences in chromatin loops regulating imprinted transcription. These results provide an integrated view of the 15q11-q13 epigenetic landscape during LUHMES neuronal differentiation, underscoring the complex interplay of transcription, chromatin looping, and DNA methylation. They also provide insights for future therapeutic approaches for AS and PWS.

Epigenetics in Prader-Willi Syndrome.

Prader-Willi Syndrome (PWS) is a rare neurodevelopmental disorder that affects approximately 1 in 20,000 individuals worldwide. Symptom progression in PWS is classically characterized by two nutritional stages. Stage 1 is hypotonia characterized by poor muscle tone that leads to poor feeding behavior causing failure to thrive in early neonatal life. Stage 2 is followed by the development of extreme hyperphagia, also known as insatiable eating and fixation on food that often leads to obesity in early childhood. Other major features of PWS include obsessive-compulsive and hoarding behaviors, intellectual disability, and sleep abnormalities. PWS is genetic disorder mapping to imprinted 15q11.2-q13.3 locus, specifically at the paternally expressed SNORD116 locus of small nucleolar RNAs and noncoding host gene transcripts. SNORD116 is processed into several noncoding components and is hypothesized to orchestrate diurnal changes in metabolism through epigenetics, according to functional studies. Here, we review the current status of epigenetic mechanisms in PWS, with an emphasis on an emerging role for SNORD116 in circadian and sleep phenotypes. We also summarize current ongoing therapeutic strategies, as well as potential implications for more common human metabolic and psychiatric disorders.

Phosphoserine inhibits neighboring arginine methylation in the RKS motif of histone H3.

The effects of phosphorylation of histone H3 at serine 10 have been studied in the context of other posttranslational modifications such as lysine methylation. We set out to investigate the impact of phosphoserine-10 on arginine-8 methylation. We performed methylation reactions using peptides based on histone H3 that contain a phosphorylated serine and compared the extent of arginine methylation with unmodified peptides. Results obtained via fluorography indicate that peptides containing a phosphorylated serine-10 inhibit deposition of methyl groups to arginine-8 residues. To further explore the effects of phosphoserine on neighboring arginine residues, we physically characterized the non-covalent interactions between histone H3 phosphoserine-10 and arginine-8 using 31P NMR spectroscopy. A salt bridge was detected between the negatively charged phosphoserine-10 and the positively charged unmodified arginine-8 residue. This salt bridge was not detected when arginine-8 was symmetrically dimethylated. Finally, molecular simulations not only confirm the presence of a salt bridge but also identify a subset of electrostatic interactions present when arginine is replaced with alanine. Taken together, our work suggests that the negatively charged phosphoserine maximizes its interactions. By limiting its exposure and creating new contacts with neighboring residues, it will inhibit deposition of neighboring methyl groups, not through steric hindrance, but by forming intrapeptide interactions that may mask substrate recognition. Our work provides a mechanistic framework for understanding the role of phosphoserine on nearby amino acid residues and arginine methylation.

Characteristics and perspectives of night-eating behaviour in a severely obese population.

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: Night-eating syndrome (NES) can be a feature of severe obesity. NES is a dysfunction of circadian rhythm and is associated with impaired sleep. WHAT THIS STUDY ADDS: Night eaters with severe obesity are more likely to be low in mood and unemployed compared with non-night eaters. Night eaters with severe obesity describe compulsive and uncontrolled eating. Research interest in night-eating syndrome (NES) has grown in recent years in line with increased rates of obesity. This study used a mixed-methods approach to investigate its characteristics in severe obesity. Eighty-one individuals (mean [standard deviation] age 44.6 [11.6] years, [body mass index] 50.0 [10.7] kg m(-2) ; 43% men) from a hospital-based UK obesity clinic were interviewed for NES based on 2003 criteria. Full and partial NES were combined into one night-eating behaviour (NEB) group (n = 31). Demographic and clinical characteristics were compared with those of non-NEB individuals (n = 50). NEB characteristics were also identified through exploratory thematic analysis of interview data. NEB individuals had lower mood (P = 0.01) and were less likely to be employed (P = 0.03). Differences in mean age and reported sleep duration were not significant. Thematic analysis of patient perceptions of NEB highlighted the potential heterogeneity of NEB development: NEB developed in childhood, adolescence and adulthood. Individuals reported long-standing and current sleep difficulties, negative affect and conflictful relationships. Night eating was solitary, compulsive and uncontrolled, and daytime eating patterns were chaotic. Accounts of awareness of night eating were conflicting. Severely obese night eaters are characterized by low mood and lack of employment. Further studies are required to explore behavioural and cognitive influences on night eating in severe obesity.

Validation of the Children's International Mucositis Evaluation Scale (ChIMES) in paediatric cancer and SCT.

BACKGROUND: Objectives were to describe the reliability and validity of a new paediatric-specific mucositis scale, the Children's International Mucositis Evaluation Scale (ChIMES). METHODS: In a multi-centre prospective study, children aged 0 to ≤18 years were eligible if they were receiving any of the following: myeloablative stem cell transplantation (SCT), ≥60 mg m(-2) course(-1) doxorubicin or ≥12 g m(-2) methotrexate. Multiple measures of mucositis were included along with ChIMES. Respondents were parent proxy report for children aged <12 years, and child self-report for children aged 12-18 years and 8 to <12 years. Mucositis diaries were completed at baseline and on Days 7-17 following chemotherapy/conditioning. On Day 14, the respondent reported presence of mucositis and change since the previous day. RESULTS: The 185 respondents included parents (N=98), children aged 12-18 years (N=66) and children aged 8 to <12 years (N=21). Test-retest reliability was excellent for ChIMES Total Score and ChIMES Percentage Score with r>0.8 for all respondent types. Criteria for construct validation were met across all measures. ChIMES also demonstrated responsiveness with significant differences between baseline and Day 14. CONCLUSION: ChIMES is a paediatric-specific measure of mucositis with favourable psychometric properties. It exhibits reliability, construct validity and responsiveness. ChIMES should be incorporated into clinical trials of mucositis prevention and treatment in paediatric cancer and SCT.

Night eating syndrome: implications for severe obesity.

Night eating syndrome (NES) was first identified in 1955 by Stunkard, a psychiatrist specialising in eating disorders (ED). Over the last 20 years considerable progress has been made in defining NES as a significant clinical entity in its own right and it has now been accepted for inclusion in the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) due for publication in 2013. NES is considered a dysfunction of circadian rhythm with a disassociation between eating and sleeping. Core criteria include a daily pattern of eating with a significantly increased intake in the evening and/or night time, as manifested by one or both of the following: at least 25% of food intake is consumed after the evening meal or at least two episodes of nocturnal eating per week. An important recent addition to core criteria includes the presence of significant distress and/or impairment in functioning. Stunkard's team recommend further investigation on the pathogenesis of NES, in particular its relationship with traumatic life events, psychiatric comorbidity, the age of onset of NES and course of NES over time. The relationship between NES and other ED also requires further clarification as night-eaters exhibit some features of other ED; previous guidance to separate NES from other ED may have hindered earlier characterisation of NES. Evidence from European and American studies suggests NES features strongly in populations with severe obesity. The complex interplay between depression, impaired sleep and obesity-related comorbidity in severely obese individuals makes understanding NES in this context even more difficult. This review examines evidence to date on the characterisation of NES and concludes by examining the applicability of current NES criteria to individuals with severe obesity.

Dental disease in type 3 Von Willebrand disease: a neglected problem.

Type 3 Von Willebrand disease (VWD) is a rare, severe, autosomal recessive bleeding disorder. In our institution, we follow 17 children with type 3 VWD. We have observed a high prevalence of dental disease in these patients prompting us to undertake a retrospective review of our cohort of patients with type 3 VWD to catalogue the extent of their dental disease. Sixteen of these patients have been assessed by our dentistry department. Five children have undergone minor dental procedures (e.g. restorations, stainless steel crowns) and seven major procedures (e.g. dental extractions, pulpotomies and root canal treatments). These patients have collectively used 85,400 (ristocetin cofactor) IU of Humate-P on dental procedures alone. In addition to the considerable costs of factor are the cost of operating room time, dentists' costs, and the cost of other topical haemostatic agents (e.g. Tisseel) used during their dental procedures. As such there is considerable morbidity and cost from dental disease in these patients that is much higher than what is seen in patients with haemophilia or in the normal paediatric population. We speculate that the combination of these patients having a significant mucosal bleeding disorder together with various socioeconomic factors contribute to the significant degree of dental disease seen in this group of patients. We would suggest that better preventive dental care needs to be provided to these patients to avoid the considerable morbidity and very high burden of dental disease in type 3 VWD.

Knowledge, compliance and serum phenylalanine concentrations in adolescents and adults with phenylketonuria and the effect of a patient-focused educational resource.

BACKGROUND: There is a lack of dedicated resources for adolescent and adult patients with phenylketonuria (PKU) and few studies have examined dietary practices within this group. METHODS: One hundred and seventy-seven PKU patients were sent questionnaires to assess dietary compliance and the preferred format for an educational resource. Seventy-one patients responded; 32 following diet were recruited to assess the resource's impact on test variables. The results were compared for the intervention group (n = 22) and control group (n = 10) at baseline, and 1 and 6 months after resource intervention. RESULTS: Most patients were aware of dietary recommendations, although this did not always result in compliance. The preferred resource format was a filofax-style folder with inserts (P < 0.05). There was a significant difference in the extent of change in knowledge score between baseline and 1 month in favour of the intervention group (P < 0.05). The improvement in knowledge was not accompanied by a significant improvement in measures of compliance. CONCLUSIONS: These findings add to the knowledge base about this patient group and support the use of patient involvement in resource development. It is likely that the test parameters used were not sensitive enough to pick up subtle and longer-term effects on compliance.

Anxiety, depression and disability across the lifespan.

The authors examined the relationship between anxiety, depression and physical disability, after controlling for demographic and health variables, in a sample of 374 adults aged 18-94. Results indicate that anxiety, depression and comorbid anxiety and depression are associated with higher levels of disability, after controlling for factors such as age, gender, income, self-rated health, number of medical conditions and number of physician visits in the past year. Furthermore, anxiety, depression and comorbid anxiety and depression have a differential effect on disability according to age, with older adults with any of these symptoms reporting higher levels of disability than younger adults. These findings suggest that physicians working with older adults should assess for and treat anxiety as well as depressive symptoms.

Quality of life in adult enteral tube feeding patients.

BACKGROUND: Enteral tube feeding may impact significantly on patients' quality of life (QoL). The aim of this paper is to review studies that have measured QoL in adult patients receiving enteral tube feeding to determine the factors that are associated with feeding that impact on the patients' QoL. METHODS: An electronic search of CINAHL, MEDLINE and EMBASE was undertaken to identify articles that had measured QoL in adults receiving enteral tube feeding. RESULTS: Ten studies were included in the review. Quality of life in patients receiving enteral tube feeding generally appeared poor compared with control groups. Factors that were found to impact on patients' quality of life included symptoms such as nausea, vomiting, diarrhoea and fatigue. Issues around body image, inability to go out and discomfort while carrying out activities of daily living also impacted on QoL. CONCLUSIONS: Although the presence of underlying disease and the use of different QoL measures made comparison of the results difficult, issues requiring further consideration in clinical practice include effective symptom management, assessment of psychological and emotional issues and support for patients to enable them to cope with the resulting social isolation.

Serial controlled N-of-1 trials of topical vitamin E as prophylaxis for chemotherapy-induced oral mucositis in paediatric patients.

The objectives were (1) to determine whether in children undergoing doxorubicin-containing chemotherapy, topical vitamin E decreases an objective measurement of oral mucositis compared to placebo, and (2) to assess the feasibility of an innovative trial design in paediatric cancer, combining N-of-1 trials using Bayesian meta-analysis. We conducted a series of N-of-1, double-blinded, randomised controlled trials in children > or = 6 years of age receiving repeated cycles of identical doxorubicin-containing chemotherapy. Each study cycle was followed by topical vitamin E (800 mg) or placebo. We enroled 16 children and 45 post chemotherapy cycles were randomised to vitamin E (N=22) or placebo (N=23). There was no difference in objective mucositis scores with a mean score of 0.2 with vitamin E and 0.3 with placebo. Topical vitamin E does not reduce doxorubicin-induced oral mucositis in children. The use of N-of-1 studies and Bayesian meta-analysis may facilitate the study of some therapies in paediatric oncology.

Assessment of fecal output in patients receiving enteral tube feeding: validation of a novel chart.

OBJECTIVE: To develop a practical, valid and reliable chart to assist in the accurate visual characterization of fecal output in patients receiving enteral tube feeding (ETF). DESIGN: A chart incorporating verbal and pictorial descriptors of fecal output was developed. Validity and reliability were assessed by a questionnaire survey of health professionals and a clinical study of patients commencing ETF. Content validity was assessed from the results of the questionnaire, construct validity by contrasting groups analysis, concurrent validity by comparison of the chart with actual fecal weight and inter-rater reliability by independent characterization of the same fecal sample by two nurses. SETTING: St George's Hospital, London, UK. SUBJECTS: In all, 35 health professionals completed a questionnaire with respect to the chart. The chart was then used to monitor fecal output in 36 patients commencing ETF for a total of 171 patient-days, during which time nurses used the chart to characterize 269 fecal samples, of which 59 were subsequently weighed. RESULTS: The results of the questionnaire suggested good content validity. The chart demonstrated statistically significant differences in fecal frequency, fecal consistency, fecal score and incidence of diarrhea for contrasting patient groups expected to have different fecal output (P < 0.05). The inter-rater reliability was almost perfect for fecal consistency (95% agreement, kappa = 0.91) and substantial for fecal weight (83%, kappa = 0.75). In all, 83% of fecal samples were assigned to the correct weight category (kappa = 0.75). CONCLUSION: The chart has good content, construct and concurrent validity and inter-rater reliability, and is suitable for both research and clinical use. SPONSORSHIP: This research was jointly supported by King's College London and Nestlé, UK.

Defining and reporting diarrhoea during enteral tube feeding: do health professionals agree?

INTRODUCTION: The criteria used to define diarrhoea during enteral tube feeding (ETF) and the interrater reliability of nursing staff reporting it in clinical practice has not been investigated. METHODS: A structured questionnaire was sent to a cohort of dietitians, specialist stroke nurses, intensive therapy unit (ITU) nurses and consultant gastroenterologists regarding their criteria for defining ETF diarrhoea. In a clinical study, nursing staff independently inspected faeces from patients receiving ETF and were asked whether they considered the patient to have diarrhoea. RESULTS: Thirty-five healthcare workers responded to the questionnaire. Faecal frequency, faecal consistency and faecal quantity were all considered important criteria in defining ETF diarrhoea. Faecal frequency was considered more important than consistency and quantity (P = 0.048), although not all professional groups agreed on the order of importance. In the clinical study, nursing staff agreed on the presence or absence of diarrhoea on 75% of occasions, with agreement being only fairly reliable (kappa = 0.48). CONCLUSION: Healthcare workers use a range of criteria to define ETF diarrhoea, but may not agree on the importance of each criteria. In order to prevent confusion between professional groups, healthcare workers should be encouraged to describe faecal characteristics rather than rely on the term 'diarrhoea'.

The role of probiotics and prebiotics in the management of diarrhoea associated with enteral tube feeding.

INTRODUCTION: Diarrhoea is a common and serious complication of enteral tube feeding, and has a range of aetiologies. Manipulation of the colonic microflora may reduce the incidence of enteral tube feeding diarrhoea via suppression of enteropathogens and production of short-chain fatty acids. Probiotics and prebiotics are commonly used during enteral tube feeding to manipulate the colonic microflora; however, their efficacy is as yet uncertain. METHODS: English-language studies investigating the pathogenesis of enteral tube feeding diarrhoea and the use of probiotics and prebiotics were identified by searching the electronic databases CINAHL, EMBASE and MEDLINE from 1980 to 2001. The bibliographies of articles obtained were searched manually. RESULTS: Only two prospective, randomized, double-blind, placebo-controlled trials have investigated the effect of a probiotic on enteral tube feeding diarrhoea; however, results are conflicting. No prospective, randomized, double-blind, placebo-controlled studies have specifically addressed the effect of a prebiotic on the incidence of enteral tube feeding diarrhoea. CONCLUSION: Theoretically, probiotics and prebiotics may be of benefit in prophylaxis against enteral tube feeding diarrhoea; however, there is currently insufficient evidence to support their routine use. Prospective, randomised, double-blind, placebo-controlled studies investigating their effect on diarrhoea are required. These observations are discussed with reference to the current literature.

Randomized trial of the effects of cholesterol-lowering dietary treatment on psychological function.

PURPOSE: Epidemiological studies have suggested that cholesterol lowering could affect psychological functioning. This study was designed to test whether cholesterol-lowering diets adversely affect mood and cognitive function.5.2 mM [198 mg/dL]) to either a low-fat diet, a Mediterranean diet, or a waiting-list control. Cholesterol levels, psychological well-being (depression, anxiety, hostility), and cognitive function were assessed at baseline, 6 weeks, and 12 weeks. RESULTS: Total serum cholesterol levels fell significantly more in the intervention groups (8.2% reduction) than in the control group (P <0.001). All three groups showed a modest improvement in psychological well-being during the 12-week treatment period, but there were no differences among the groups. There were no between-group differences on three measures of cognitive function, but for a fourth measure, which involved the task with the greatest processing load, the two intervention groups did significantly worse (P <0.001) than the control group. The change in performance was correlated with the change in total serum cholesterol level (r = 0. 21, P = 0.01). CONCLUSIONS: Two dietary interventions that successfully lowered serum cholesterol levels had no adverse effect on mood. There was some evidence for a relative impairment in cognitive function in the treated groups in one of four cognitive tests, but additional studies will be required to determine the relevance of this finding.

Frequency of consumption and nutrient composition of composite dishes commonly consumed in the UK by South Asian Muslims originating from Bangladesh, Pakistan and East Africa (Ismailis).

INTRODUCTION: This paper presents information on the nutrient composition of commonly consumed traditional dishes eaten by the three major South Asian Muslim groups residing in Britain, namely Bangladeshi, Pakistani and East African Ismaili Muslims. METHODS: Information regarding the most common dishes consumed by South Asian Muslims originating from Bangladesh, Pakistan and East Africa (Ismailis) and living in London was obtained from 7-day menu records over two seasons. For each common dish, weighed recipes were collected in triplicate and the composition (energy and selected nutrients) was calculated from the ingredients and cooked weight of the dish. RESULTS: The three Muslim groups showed considerable variation in traditional foods commonly consumed as well as variation in fat and energy contents of similar recipes both within and between groups. Nutrient composition of commonly consumed dishes is presented calculated from the recipe nearest the average in terms of fat and energy for a particular dish. DISCUSSION AND CONCLUSIONS: The potential uses of the data, one of which would be to improve dietary compliance (especially to lower fat intakes) amongst the three diverse South Asian Muslim groups, are discussed.

Antimicrobial treatment options in the management of odontogenic infections.

Most acute orofacial infections are of odontogenic origin. In normal hosts, however, they usually do not occur without some type of predisposing condition. Early recognition and management of acute orofacial infections is critical, because rapid systemic involvement can occur, especially in children. Antimicrobial therapy has an essential role in the management of these infections. If it is initiated before surgery, it can shorten the period of infection and minimize associated risks. The etiology of odontogenic infections is usually attributed to the endogenous flora of the mouth, and not to the introduction of non-resident bacteria. Odontogenic infections are typically polymicrobial; however, anaerobes generally outnumber aerobes by at least four fold. The penicillins have historically been used as the first-line therapy in these cases, but increasing rates of resistance have lowered their usefulness. Bacterial resistance to this class of agents is predominately achieved through the production of beta-lactamases. Clindamycin, because of its broad spectrum of activity and resistance to beta-lactamase degradation, is an attractive first-line therapy in the treatment of odontogenic infections.

African plant foods rich in non-starch polysaccharides reduce postprandial blood glucose and insulin concentrations in healthy human subjects.

The effects of two vegetable flours, prepared from the African plants Detarium senegalense Gmelin, a legume, and Cissus rotundifolia, a shrub, on postprandial blood glucose and insulin concentrations in human subjects, were investigated. Chemical analysis indicated that these flours contained significant amounts of NSP. The detarium in particular was found to be a rich source of water-soluble NSP (SNSP). The flours were incorporated into two types of breakfast meal, a stew meal and a wheat bread meal, containing 50 g and 70 g available carbohydrate respectively. Both meals also contained 10-12 g NSP, the major fraction of which was SNSP. Control and fibre-rich meals were consumed on separate days in randomized order by two different groups of subjects (n 5, stew meals; n 10, bread meals). Venous blood samples were taken at fasting (0 min) and postprandially at 30 min intervals for 2.5 h and the plasma analysed for glucose and insulin. Compared with the controls, detarium and cissus meals elicited significant reductions (P < 0.006) in plasma glucose levels at most postprandial time points and for area-under-the-curve (AUC) values (AUC reductions 38-62%). Significant reductions (P < 0.002) in plasma insulin levels at various postprandial time points and for AUC values were also seen after detarium and cissus breads (AUC reductions 43 and 36% respectively), but not after the fibre-rich stew meals. SNSP and starch are possibly the main, but not the only, components responsible for the glucose- and insulin-lowering effects of cissus flour. The main SNSP fraction of detarium, identified as a high-molecular-weight xyloglucan, is likely to be a primary factor in determining the physiological activity of detarium flour.