Effect of video angle on detection of induced front limb lameness in horses.

BACKGROUND: Lameness examinations are commonly performed in equine medicine. Advancements in digital technology have increased the use of video recordings for lameness assessment, however, standardization of ideal video angle is not available yielding videos of poor diagnostic quality. The objective of this study was to evaluate the effect of video angle on the subjective assessment of front limb lameness. A randomized, blinded, crossover study was performed. Six horses with and without mechanically induced forelimb solar pain were recorded using 9 video angles including horses trotting directly away and towards the video camera, horses trotting away and towards a video camera placed to the left and right side of midline, and horses trotting in a circle with the video camera placed on the inside and outside of the circle. Videos were randomized and assessed by three expert equine veterinarians using a 0-5 point scoring system. Objective lameness parameters were collected using a body-mounted inertial sensor system (Lameness Locator®, Equinosis LLC). Interobserver agreement for subjective lameness scores and ease of grading scores were determined. RESULTS: Induction of lameness was successful in all horses. There was excellent agreement between objective lameness parameters and subjective lameness scores (AUC of the ROC = 0.87). For horses in the "lame" trials, interobserver agreement was moderate for video angle 2 when degree of lameness was considered and perfect for video angle 2 and 9 when lameness was considered as a binary outcome. All other angles had no to fair agreement. For horses in the "sound" trials, interobserver agreement was perfect for video angle 5. All other video angles had slight to moderate agreement. CONCLUSIONS: When video assessment of forelimb lameness is required, a video of the horse trotting directly towards the video camera at a minimum is recommended. Other video angles may provide supportive information regarding lameness characteristics.

Cord blood DNA methylation of immune and lipid metabolism genes is associated with maternal triglycerides and child adiposity.

OBJECTIVE: Fetal exposures may impact offspring epigenetic signatures and adiposity. The authors hypothesized that maternal metabolic traits associate with cord blood DNA methylation, which, in turn, associates with child adiposity. METHODS: Fasting serum was obtained in 588 pregnant women (27-34 weeks' gestation), and insulin, glucose, high-density lipoprotein cholesterol, triglycerides, and free fatty acids were measured. Cord blood DNA methylation and child adiposity were measured at birth, 4-6 months, and 4-6 years. The association of maternal metabolic traits with DNA methylation (429,246 CpGs) for differentially methylated probes (DMPs) and regions (DMRs) was tested. The association of the first principal component of each DMR with child adiposity was tested, and mediation analysis was performed. RESULTS: Maternal triglycerides were associated with the most DMPs and DMRs of all traits tested (261 and 198, respectively, false discovery rate < 0.05). DMRs were near genes involved in immune function and lipid metabolism. Triglyceride-associated CpGs were associated with child adiposity at 4-6 months (32 CpGs) and 4-6 years (2 CpGs). One, near CD226, was observed at both timepoints, mediating 10% and 22% of the relationship between maternal triglycerides and child adiposity at 4-6 months and 4-6 years, respectively. CONCLUSIONS: DNA methylation may play a role in the association of maternal triglycerides and child adiposity.

Incidence and risk factors for complications associated with equine general anaesthesia for elective magnetic resonance imaging.

BACKGROUND: Complications are a practical consideration for elective magnetic resonance imaging (MRI) studies performed under general anaesthesia but relatively little is known about their distribution and risk factors. OBJECTIVES: To describe the incidence of complications associated with MRI performed under general anaesthesia at a large referral facility and evaluate potential risk factors for these complications. STUDY DESIGN: Retrospective case-control study. METHODS: Patient information and details of the MRI procedure were collected retrospectively from medical records of all horses that had undergone an MRI under general anaesthesia at the University of Pennsylvania, New Bolton Center, between September 2005 and April 2012. Complications and categorical variables were examined by chi-squared or Fisher's exact tests as appropriate. A mixed-effects logistic regression approach was used to evaluate associations between explanatory variables and the outcome variable (complications or pyrexia). A univariable screen was used to select variables (likelihood ratio test p < 0.2) for inclusion in the multivariable analysis. Statistical significance was inferred when p ≤ 0.05. RESULTS: Complications were noted after MRI in 51 (17.4%) of 293 events eligible for inclusion. Complications included pyrexia (n = 35), pneumonia (n = 14), colic (n = 10), facial/nerve paralysis (n = 6), diarrhoea (n = 4), and other (n = 3). The odds of developing a post-anaesthetic complication were significantly decreased in horses that received peri-anaesthetic antimicrobials (OR 0.29, 95% CI 0.14-0.63, p = 0.002). Increased age (OR 0.87, 95% CI, 0.76-0.99, p = 0.03) and peri-anaesthetic antimicrobial administration (OR 0.23, 95% CI 0.08-0.65, p = 0.005) were associated with a decreased odds of developing pyrexia. MAIN LIMITATIONS: Single centre retrospective design. CONCLUSIONS: Potential complications including pyrexia, pneumonia and colic should be recognised when pursuing MRI under general anaesthesia. The administration of peri-anaesthetic antimicrobials decreased the odds of a complication and warrants consideration, particularly in horses that might be classified as high risk.

Interobserver agreement of lateral and oblique radiography and standing cone beam CT of the caudal cervical articular process joints of horses.

Caudal cervical articular process joint osteoarthritis (CAPJ OA) leads to career-altering clinical signs in the horse. Oblique radiographs and standing cone beam computed tomography (CBCT) facilitate the assessment of this area, however, the variability of interpretation of these images is currently unknown. This retrospective, secondary analysis, methods comparison study investigated interobserver agreement between clinicians and modality in grades of CAPJ OA on lateral and oblique radiographs and CBCT. We hypothesized that agreement between clinicians' grades of CAPJ OA would be lowest for oblique radiographs and highest for CBCT, and agreement between grades of CAPJ OA would be low for all pairs of modalities. Horses underwent lateral and oblique radiography and CBCT of the CAPJs of C5-C6 and C6-C7. Radiographs and CBCT images were graded retrospectively by four blinded clinicians using 3-point scales. Cohen's kappa analysis was used to evaluate interobserver agreement between grades of CAPJ OA, and agreement between grades of CAPJ OA between different modalities was explored using kappa-weighted analysis. Agreement between clinicians' grades of CAPJ OA was moderate for lateral radiographs (0.49), and fair for oblique radiographs (0.23) and CBCT (0.36). For all modalities, agreement was slight to fair between clinicians for CAPJs with grade 1 (normal, 0.21-0.32) or 2 (mild, 0.13-0.36) CAPJ OA, and moderate to substantial for grade 3 (moderate to severe, 0.45-0.77) CAPJ OA. Agreement between grades of CAPJ OA was fair for all pairs of modalities. This study provides important information regarding the inconsistency of interpretation of mild CAPJ OA on radiographs and CBCT amongst clinicians.

Sex-based differences in placental DNA methylation profiles related to gestational age: an NIH ECHO meta-analysis.

The placenta undergoes many changes throughout gestation to support the evolving needs of the foetus. There is also a growing appreciation that male and female foetuses develop differently in utero, with unique epigenetic changes in placental tissue. Here, we report meta-analysed sex-specific associations between gestational age and placental DNA methylation from four cohorts in the National Institutes of Health (NIH) Environmental influences on Child Health Outcomes (ECHO) Programme (355 females/419 males, gestational ages 23-42 weeks). We identified 407 cytosine-guanine dinucleotides (CpGs) in females and 794 in males where placental methylation levels were associated with gestational age. After cell-type adjustment, 55 CpGs in females and 826 in males were significant. These were enriched for biological processes critical to the immune system in females and transmembrane transport in males. Our findings are distinct between the sexes: in females, associations with gestational age are largely explained by differences in placental cellular composition, whereas in males, gestational age is directly associated with numerous alterations in methylation levels.

Multiomic Signatures of Chronic Beryllium Disease Bronchoalveolar Lavage Cells Relate to T-Cell Function and Innate Immunity.

Chronic beryllium disease (CBD) is a Th1 granulomatous lung disease preceded by sensitization to beryllium (BeS). We profiled the methylome, transcriptome, and selected proteins in the lung to identify molecular signatures and networks associated with BeS and CBD. BAL cell DNA and RNA were profiled using microarrays from CBD (n = 30), BeS (n = 30), and control subjects (n = 12). BAL fluid proteins were measured using Olink Immune Response Panel proteins from CBD (n = 22) and BeS (n = 22) subjects. Linear models identified features associated with CBD, adjusting for covariation and batch effects. Multiomic integration methods identified correlated features between datasets. We identified 1,546 differentially expressed genes in CBD versus control subjects and 204 in CBD versus BeS. Of the 101 shared transcripts, 24 have significant cis relationships between gene expression and DNA methylation, assessed using expression quantitative trait methylation analysis, including genes not previously identified in CBD. A multiomic model of top DNA methylation and gene expression features demonstrated that the first component separated CBD from other samples and the second component separated control subjects from remaining samples. The top features on component one were enriched for T-lymphocyte function, and the top features on component two were enriched for innate immune signaling. We identified six differentially abundant proteins in CBD versus BeS, with two (SIT1 and SH2D1A) selected as important RNA features in the multiomic model. Our integrated analysis of DNA methylation, gene expression, and proteins in the lung identified multiomic signatures of CBD that differentiated it from BeS and control subjects.

Associations between child filaggrin mutations and maternal diet with the development of allergic diseases in children.

BACKGROUND: Filaggrin (FLG) loss-of-function mutations in children and maternal diet in pregnancy have been implicated in child allergy outcomes. This paper studies the questions: "do FLG mutations modify the effect of maternal diet on the odds of development of allergic diseases?" and "which factor leads to the highest rate of diagnosis allergic diseases over time, maternal diet, or FLG mutations?". METHODS: Exact logistic regressions studied effect modification. Cox proportional hazard models compared the rate of allergic disease development in three groups (N = 624): (1) children with FLG mutation, (2) children without FLG mutation whose mothers did not eat an allergy preventive diet, and (3) children without FLG mutation whose mothers ate an allergy preventive diet. Maternal diet was classified using a validated index. RESULTS: Cox models showed the development of atopic dermatitis, asthma, and wheeze was significantly higher for children in group 1 versus 3 (HR = 2.40 [1.32, 4.37], HR = 2.29 [1.05, 4.97], and HR 2.10 [1.004, 4.38], respectively), but not significantly higher for children in group 1 versus 2 (HR = 1.30 [0.74, 2.29], HR = 1.27 [0.61, 2.63], and HR = 1.29 [0.65, 2.58], respectively). Development of allergic rhinitis was significantly higher for group 1 versus 2 and 3 (1 vs. 2: HR = 2.29 [1.10, 4.76]; 1 vs. 3: HR = 3.21 [1.46, 7.08]). There was no significant effect modification for any outcome. CONCLUSION: Children with FLG mutation had similar risk of atopic dermatitis, asthma, and wheeze as children without an FLG mutation whose mothers did not eat an allergy preventive diet during pregnancy. Child FLG mutation did not modify the effect of maternal diet. The results suggest that maternal diet in pregnancy, a modifiable risk factor, could be a target for preventive interventions.

Treatment and outcome of eight horses with limb cellulitis and septic tendonitis or desmitis.

OBJECTIVE: To report the clinical features, treatment, and outcome in horses with cellulitis and concurrent septic tendonitis and/or desmitis. STUDY DESIGN: Short case series. METHODS: Medical records from 2000 to 2019 were reviewed, identifying horses with cellulitis and concurrent septic tendonitis and/or desmitis based on sonographic examination and positive bacterial culture. Signalment, ultrasonographic results, bacterial culture, treatment, duration of hospitalization, and complications were recorded. Long-term outcome data were obtained from follow-up examinations and/or telephone interviews. Successful outcome was defined as return to intended use. RESULTS: Eight horses met the inclusion criteria. All infections occurred in hindlimbs, with septic suspensory ligament in six of eight horses, and septic superficial digital flexor tendon in one of eight horses. Surgical debridement was performed in six of eight horses. All horses were treated with systemic and regional intravenous antimicrobials and were discharged from the hospital. Long-term follow-up was available in seven of eight horses. Of these, four horses returned to their intended athletic function, two horses returned to their intended function as a broodmare or pasture pet, and one horse is still rehabilitating. CONCLUSION: Septic tendonitis or desmitis is a rare but possible sequela of limb cellulitis. Based on the findings in this study, prognosis for return to athletic function is fair to good for horses diagnosed with cellulitis and concurrent septic tendonitis or desmitis.

Preconceptional Lipid-Based Nutrient Supplementation in 2 Low-Resource Countries Results in Distinctly Different IGF-1/mTOR Placental Responses.

BACKGROUND: Preconceptional maternal small-quantity lipid-based nutrient supplementation (SQLNS) improved intrauterine linear growth in low-resource countries as demonstrated by the Women First Preconception Maternal Nutrition Trial (WF). Fetal growth is dependent on nutrient availability and regulated by insulin-like growth factor 1 (IGF-1) through changes in placental transfer capacity, mediated by the mechanistic target of rapamycin (mTOR) pathway. OBJECTIVES: Our objective was to evaluate the role of placental mTOR and IGF-1 signaling on fetal growth in women from 2 low-resource countries with high rates of stunting after they received preconceptional SQLNS. METHODS: We studied 48 women from preconception through delivery who were from Guatemala and Pakistan and received SQLNS or not, as part of the WF study. Placental samples were obtained at delivery (control, n = 24; SQLNS, n = 24). Placental protein or mRNA expression of eukaryotic translation initiation factor binding protein-1 (4E-BP1), ribosomal protein S6 (rpS6), AMP-activated protein kinase α (AMPKA), IGF-1, insulin-like growth factor receptor (IGF-1R), and pregnancy associated plasma protein (PAPP)-A, and DNA methylation of the IGF1 promoter were determined. Maternal serum IGF-1, insulin-like growth factor binding protein (IGFBP)-3, IGFBP-4, IGFBP-5, PAPP-A, PAPP-A2, and zinc were measured. RESULTS: Mean ± SEM maternal prepregnancy BMI differed between participants in Guatemala (26.5 ± 1.3) and Pakistan (19.8 ± 0.7) (P < 0.001). In Pakistani participants, SQLNS increased the placental rpS6(T37/46):rpS6 ratio (1.5-fold) and decreased the AMPKA(T172):AMPKA ratio. Placental IGF1 mRNA expression was positively correlated with birth length and birth weight z-scores. Placental PAPP-A (30-fold) and maternal serum zinc (1.2-fold) increased with SQLNS. In Guatemalan participants SQLNS did not influence placental mTOR signaling. Placental IGF-1R protein expression was positively associated with birth length and birth weight z-scores. SQLNS increased placental PAPP-A (40-fold) and maternal serum IGFBP-4 (1.6-fold). CONCLUSIONS: In Pakistani pregnant women with poor nutritional status, preconceptional SQLNS activated placental mTOR and IGF-1 signaling and was associated with improved fetal growth. In contrast, in Guatemalan women SQLNS did not activate placental nutrient-sensing pathways. In populations experiencing childhood stunting, preconceptional SQLNS improves placental function and fetal growth only in the context of poor maternal nutrition. This trial was registered at clinicaltrials.gov as NCT01883193.

Inflammatory cytokines in horses with cervical articular process joint osteoarthritis on standing cone beam computed tomography.

BACKGROUND: Standing cone beam computed tomography (CT) provides cross-sectional imaging of the caudal cervical articular process joints (CAPJs) in the sedated horse, though the clinical implications of osteoarthritis (OA) identified on CT in this location are unknown. Increases in concentrations of intra-synovial cytokines could lend support to the clinical significance of CAPJ OA identified on this imaging modality. OBJECTIVES: Investigate the presence and concentration of intra-synovial inflammatory cytokines in CAPJs with and without standing cone beam CT evidence of OA using an equine specific multiplex assay. STUDY DESIGN: Prospective clinical study. METHODS: Standing cone beam CT of C5-6 and C6-7 was performed on horses with CAPJ OA and control horses. Synovial fluid samples of the CAPJs of C5-6 and C6-7 were obtained bilaterally using ultrasound guidance and analysed for concentrations of IFN-γ, IL-1ß, IL-6, IL-10, IL-17 and TNFα with the Milliplex® multi-analyte profiling kit. CT Images were retrospectively graded using a novel grading scheme. Significant differences between concentrations of inflammatory cytokines between joints with different categories of osteoarthritis severity were explored using a Wilcoxon rank-sum test or Kruskal-Wallis test. RESULTS: Concentrations of intra-synovial cytokines were higher in joints with moderate to severe OA when compared to joints with no or mild OA, with differences in concentrations of IL-17 reaching statistical significance (P = .007). MAIN LIMITATIONS: Limitations include discrepancy in number, age, and breed between control and OA populations, use of a novel grading scheme, and lack of a histologic gold-standard to confirm the presence and severity of CAPJ OA. CONCLUSIONS: Differences in inflammatory cytokines between caudal CAPJs with and without evidence of moderate to severe osteoarthritis on standing cone beam CT exist. This finding lends support to the clinical relevance of a diagnosis of moderate to severe CAPJ OA in the caudal cervical vertebral column as identified with this imaging modality.

Integrating family and friend support, information technology, and diabetes education in community-centric diabetes self-management.

OBJECTIVE: Diabetes self-management (DSM) education, social support, and information technology interventions can improve patient engagement and health. A major challenge is animating, integrating, and accessing resources in under-resourced, rural communities. Set in an island community in Hawai'i, this study piloted a program that integrated friend-and-family support, community health services, telehealth-enabled DSM education, and mobile technologies by activating the community's social capital to support the program. MATERIALS AND METHODS: An action research approach informed the design and implementation of a community-based DSM program that included: friends and family support, telehealth classes, personalized consultations, Bluetooth-enabled blood glucose monitors, and text messaging support. Outcomes were evaluated using biometric data, surveys, interviews, and participant observations. RESULTS: The study spanned 9 months with 7 dyads, each with 1 individual with type 2 diabetes and a friend or family member. Six of the 7 participants with diabetes experienced reduced hemoglobin A1c percentages, with 3 reducing by more than 1%. The seventh participant maintained a hemoglobin A1c level within American Diabetes Association recommended ranges. DSM knowledge and self-care behaviors improved overall. Interviews and participant observations highlighted program strengths and social challenges associated with the interpersonal relationships between the members of the dyads. CONCLUSIONS: A community-centric diabetes program can enhance understanding of diabetes etiology, DSM activities, and communication skills for effective disease management support in under-resourced rural communities. Social capital among community members, leveraged with health information technology, can catalyze and integrate limited health system resources for DSM and social support as a cost-effective strategy to develop community-centric chronic healthcare management initiatives.

Long-term outcome and effect of diagnostic analgesia in horses undergoing interspinous ligament desmotomy for overriding dorsal spinous processes.

OBJECTIVE: To report the long-term outcome of horses treated with interspinous ligament desmotomy (ISLD) for pain associated with overriding dorsal spinous processes (ORDSP) and determine the influence of preoperative diagnostic analgesia on long-term outcome. STUDY DESIGN: Retrospective study. ANIMALS: Eighteen horses. METHODS: Data were collected from horses presenting for ISLD to the University of Pennsylvania New Bolton Center between January 2013 and May 2018. Follow-up of ≥3 months postsurgically was obtained from the owner, trainer, or referring veterinarian. Long-term improvement was compared between horses that improved with diagnostic analgesia presurgically and horses that did not undergo diagnostic analgesia presurgically by using a χ2 test. Univariate logistic regression was used to test associations between long-term improvement and independent variables. RESULTS: Clinical signs had improved in 13 of 18 horses at long-term follow-up (median, 14.5 months; range, 3-57). Clinical signs improved in nine of 10 horses responding to diagnostic analgesia but only in four of eight horses that did not undergo diagnostic analgesia (χ2 [1], N = 18) = 3.55; P = .06). Although the likelihood of long-term improvement increased with prior diagnostic analgesia (odds ratio = 6.3; 95% confidence interval = 0.73, 55.0; P = .09), it did not reach statistical significance. CONCLUSION: A higher proportion of horses experienced long-term improvement in clinical signs after ISLD when horses responding to preoperative diagnostic analgesia were compared with horses that were not tested. CLINICAL SIGNIFICANCE: This study provides some evidence to support the use of diagnostic analgesia in conjunction with clinical examination for identification of clinically relevant ORDSP.

Effects of acepromazine and xylazine on subjective and objective assessments of forelimb lameness.

BACKGROUND: To facilitate lameness evaluation, sedatives such as xylazine and acepromazine are regularly used in the clinical setting, despite concerns that they may confound lameness assessment. OBJECTIVES: The objective of this study was to determine the effect of low doses of acepromazine and xylazine on subjective and objective lameness assessment. STUDY DESIGN: Randomised, blinded, crossover study. METHODS: Six horses with experimentally induced solar pain were evaluated over a 1-hour period after treatment with intravenous xylazine (0.1 or 0.2 mg/kg), intravenous acepromazine (0.02 or 0.04 mg/kg), intravenous saline (1 mL) or local analgesia (4 mL 2% mepivacaine administered subcutaneously). Lameness was assessed objectively with inertial sensors and subjectively on a scale from 0 to 5. Lameness assessments were compared with logistic regression analysis to account for the repeated measures and cross-over study design (P < .05). RESULTS: Xylazine (0.1 and 0.2 mg/kg) or acepromazine (0.02 and 0.04 mg/kg) did not result in significant differences in objective lameness assessment (vector sum) or average subjective lameness grade. Local analgesia was associated with a decrease in subjective lameness grade (OR 0.32 [0.11-0.92], P = .03). Objective measures of lameness (vector sum) were significantly decreased 45 minutes (vector sum 41.8, P = .04) and 60 minutes (vector sum 47.3, P = .03) following local analgesia administration compared with baseline (vector sum 121.4). MAIN LIMITATIONS: Extrapolation of the experimental model of moderate lameness used in this study to broad range of clinical lameness situations should be performed carefully. CONCLUSIONS: These results support the use of low doses of xylazine or acepromazine to facilitate forelimb lameness evaluation up to 1 hour in duration.

DNA Methylation Changes in Lung Immune Cells Are Associated with Granulomatous Lung Disease.

Epigenetic marks are likely to explain variability of response to antigen in granulomatous lung disease. The objective of this study was to identify DNA methylation and gene expression changes associated with chronic beryllium disease (CBD) and sarcoidosis in lung cells obtained by BAL. BAL cells from CBD (n = 8), beryllium-sensitized (n = 8), sarcoidosis (n = 8), and additional progressive sarcoidosis (n = 9) and remitting (n = 15) sarcoidosis were profiled on the Illumina 450k methylation and Affymetrix/Agilent gene expression microarrays. Statistical analyses were performed to identify DNA methylation and gene expression changes associated with CBD, sarcoidosis, and disease progression in sarcoidosis. DNA methylation array findings were validated by pyrosequencing. We identified 52,860 significant (P < 0.005 and q < 0.05) CpGs associated with CBD; 2,726 CpGs near 1,944 unique genes have greater than 25% methylation change. A total of 69% of differentially methylated genes are significantly (q < 0.05) differentially expressed in CBD, with many canonical inverse relationships of methylation and expression in genes critical to T-helper cell type 1 differentiation, chemokines and their receptors, and other genes involved in immunity. Testing of these CBD-associated CpGs in sarcoidosis reveals that methylation changes only approach significance, but are methylated in the same direction, suggesting similarities between the two diseases with more heterogeneity in sarcoidosis that limits power with the current sample size. Analysis of progressive versus remitting sarcoidosis identified 15,215 CpGs (P < 0.005 and q < 0.05), but only 801 of them have greater than 5% methylation change, demonstrating that DNA methylation marks of disease progression changes are more subtle. Our study highlights the significance of epigenetic marks in lung immune response in granulomatous lung disease.

Lameness Evaluation of the Athletic Horse.

Lameness examination is commonly performed in the athletic horse. A skilled lameness diagnostician must have keen clinical and observational skills. Evaluation starts with a detailed history and thorough physical examination. Next, gait evaluation in the moving horse is performed. Lame horses have asymmetrical body movement due to unconscious shift of body weight. Recognition of the resultant head nod and pelvic hike is the basis for lameness diagnosis. Lameness identification is enhanced by circling, limb flexions, and riding. Most lame horses do not exhibit pathognomonic gait characteristics, and therefore, diagnostic analgesia is the best way to authenticate underlying sites of pain.

Role of epigenetics in the development of childhood asthma.

PURPOSE OF REVIEW: Epigenetic marks are emerging as mediators of genetics and the environment on complex disease phenotypes, including childhood asthma and allergy. RECENT FINDINGS: Epigenome-wide association studies over the past year have added to the growing body of evidence supporting significant associations of epigenetic regulation of gene expression and asthma and allergy. Studies in children have identified signatures of eosinophils in peripheral blood, Th2 cell transcription factors and cytokines in peripheral blood mononuclear cells, and epithelial dysfunction in the respiratory epithelium. Importantly, studies at birth have begun to decipher the contribution of epigenetic marks to asthma inception. Few studies have also begun to address the contribution of genetics and the environment to these associations. SUMMARY: Next generation of epigenome-wide association studies that will deal with confounders, study the influence of the genetics and environment, and incorporate multiple datasets to provide better interpretation of the findings are on the horizon. Identification of key epigenetic marks that are shaped by genetics and the environment, and impact transcription of specific genes will help us have a better understanding of etiology, heterogeneity and severity of asthma, and will also empower us to develop biologically driven therapeutics and biomarkers for secondary prevention of this disease.

Forkhead box protein 3 demethylation is associated with tolerance induction in peanut-induced intestinal allergy.

BACKGROUND: Regulatory T (Treg) cells play an essential role in the maintenance of immune homeostasis in allergic diseases. OBJECTIVES: We sought to define the mechanisms underlying induction of tolerance to peanut protein and prevention of the development of peanut allergy. METHODS: High or low doses of peanut extract were administered to pups every day for 2 weeks before peanut sensitization and challenge. After challenge, symptoms, Treg cell numbers, and forkhead box protein 3 (Foxp3), TH2 and TH17 cytokine, and Tgfß expression in mesenteric lymph node (MLN) CD4+ T cells and jejunum were monitored. Treg cell suppressive activity and Foxp3 methylation in MLN CD4+ T cells were assayed. RESULTS: Feeding high but not low doses of peanut before sensitization induced tolerance, as demonstrated by prevention of diarrhea and peanut-specific IgE responses, increases in the percentage of CD4+CD25+FoxP3+ cells in MLNs, and Foxp3 mRNA and protein expression in CD4+ cells from MLNs or jejunum. Feeding high doses of peanut before sensitization decreased percentages of CD3+CD4+IL-13+ and CD3+CD4+IL-17+ cells in MLNs and decreased Il13 and Il17a and increased Tgfß mRNA expression in the jejunum; numbers of CD103+ dendritic cells in MLNs were significantly increased. Treg cell suppression was shown to be antigen specific. Foxp3 methylation was increased in peanut extract-sensitized and challenged mice, whereas in tolerized mice levels were significantly reduced. CONCLUSIONS: Feeding high doses of peanut to pups induced tolerance to peanut protein. Foxp3 demethylation was associated with tolerance induction, indicating that Treg cells play an important role in the regulation of peanut sensitivity and maintenance of immune homeostasis.

Maternal obesity alters fatty acid oxidation, AMPK activity, and associated DNA methylation in mesenchymal stem cells from human infants.

OBJECTIVE: Infants born to mothers with obesity have greater adiposity, ectopic fat storage, and are at increased risk for childhood obesity and metabolic disease compared with infants of normal weight mothers, though the cellular mechanisms mediating these effects are unclear. METHODS: We tested the hypothesis that human, umbilical cord-derived mesenchymal stem cells (MSCs) from infants born to obese (Ob-MSC) versus normal weight (NW-MSC) mothers demonstrate altered fatty acid metabolism consistent with adult obesity. In infant MSCs undergoing myogenesis in vitro, we measured cellular lipid metabolism and AMPK activity, AMPK activation in response to cellular nutrient stress, and MSC DNA methylation and mRNA content of genes related to oxidative metabolism. RESULTS: We found that Ob-MSCs exhibit greater lipid accumulation, lower fatty acid oxidation (FAO), and dysregulation of AMPK activity when undergoing myogenesis in vitro. Further experiments revealed a clear phenotype distinction within the Ob-MSC group where more severe MSC metabolic perturbation corresponded to greater neonatal adiposity and umbilical cord blood insulin levels. Targeted analysis of DNA methylation array revealed Ob-MSC hypermethylation in genes regulating FAO (PRKAG2, ACC2, CPT1A, SDHC) and corresponding lower mRNA content of these genes. Moreover, MSC methylation was positively correlated with infant adiposity. CONCLUSIONS: These data suggest that greater infant adiposity is associated with suppressed AMPK activity and reduced lipid oxidation in MSCs from infants born to mothers with obesity and may be an important, early marker of underlying obesity risk.

The nasal methylome and childhood atopic asthma.

BACKGROUND: Given the strong environmental influence on both epigenetic marks and allergic asthma in children, the epigenetic alterations in respiratory epithelia might provide insight into allergic asthma. OBJECTIVE: We sought to identify DNA methylation and gene expression changes associated with childhood allergic persistent asthma. METHODS: We compared genomic DNA methylation patterns and gene expression in African American children with persistent atopic asthma (n = 36) versus healthy control subjects (n = 36). Results were validated in an independent population of asthmatic children (n = 30) by using a shared healthy control population (n = 36) and in an independent population of white adult atopic asthmatic patients (n = 12) and control subjects (n = 12). RESULTS: We identified 186 genes with significant methylation changes, differentially methylated regions or differentially methylated probes, after adjustment for age, sex, race/ethnicity, batch effects, inflation, and multiple comparisons. Genes differentially methylated included those with established roles in asthma and atopy and genes related to extracellular matrix, immunity, cell adhesion, epigenetic regulation, and airflow obstruction. The methylation changes were substantial (median, 9.5%; range, 2.6% to 29.5%). Hypomethylated and hypermethylated genes were associated with increased and decreased gene expression, respectively (P < 2.8 × 10-6 for differentially methylated regions and P < 7.8 × 10-10 for differentially methylated probes). Quantitative analysis in 53 differentially expressed genes demonstrated that 32 (60%) have significant methylation-expression relationships within 5 kb of the gene. Ten loci selected based on the relevance to asthma, magnitude of methylation change, and methylation-expression relationships were validated in an independent cohort of children with atopic asthma. Sixty-seven of 186 genes also have significant asthma-associated methylation changes in nasal epithelia of adult white asthmatic patients. CONCLUSIONS: Epigenetic marks in respiratory epithelia are associated with allergic asthma and gene expression changes in inner-city children.