Development of Formaldehyde dehydrogenase-coupled assay and Antibody-based assays for ALKBH5 Activity Evaluation

Publication date: Available online 7 September 2018Source: Journal of Pharmaceutical and Biomedical AnalysisAuthor(s): Dan-Dan Shen, Feng-Zhi Suo, Qi-Meng Song, Jiao Chang, Ting Zhang, Jing-Jing Hong, Yi-Chao Zheng, Hong-Min LiuAbstractN6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic messenger RNA (mRNA). Until now, two RNA demethylases have been identified, including FTO (fat mass and obesity-associated protein) and ALKBH5 (α-ketoglutarate-dependent dioxygenase alkB homologue 5). As a mammalian m6A demethylase, ALKBH5 significantly affects mRNA export and RNA metabolism as well as the assembly of mRNA processing factors in nuclear speckles, and ALKBH5 may play a significant role in these biological processes. Nevertheless, no modulator of ALKBH5 has been reported. The reason for that may be the lack of in vitro assays for ALKBH5 inhibitor screening. Herein, we describe the development of two homogeneous assays for ALKBH5 using N6-methyladenosine as substrate with different principles. Using ALKBH5 recombinant, we developed a formaldehyde dehydrogenase coupled fluorescence based assay and an antibody based assay for the activity evaluation of ALKBH5. These robust coupled assays are suitable for screening ALKBH5 inhibitors in 384-well format (Z’ factors of 0.74), facilitating the discovery of modulators in the quest for the regulation of biological processes.
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research

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Conclusions: The results evidenced the role of the domiciliary situation among outcomes at the national level, with lower prevalence of general and abdominal obesity in men living in rural areas. However, higher prevalences were found among women, especially for abdominal obesity.
Source: Revista Brasileira de Epidemiologia - Category: Epidemiology Source Type: research
Trust for America's Health reported rising levels of obesity contributed to increased disease rates and health care costs (Levi et al., 2013). Factors associated with overweight and obesity rates include lower socioeconomic status, public insurance, and increased chronic disease rates.
Source: Journal of Pediatric Health Care - Category: Pediatrics Authors: Source Type: research
In conclusion, our results showed that GSF prevented diet-induced obesity in C57BL/6J mice, and suggested that the mechanism responsible was related to the promotion of energy metabolism.Graphical abstract
Source: Journal of Functional Foods - Category: Nutrition Source Type: research
In this study, we investigated anti-diabetic activities of a phenolic-rich extract from Hypericum attenuatum Choisy (HA-PRE). In vivo, 4-week treatment of HA-PRE improved hyperglycemia, obesity, glucose tolerance, and hyperinsulinism. Western blot analysis revealed that HA-PRE increased AMPKα and Akt phosphorylation in skeletal muscle, liver and white adipose tissue (WAT); increased GSK3β phosphorylation in skeletal muscle and liver; and enhanced GLUT4 expression in WAT and skeletal muscle. HA-PRE reduced PPARγ expression, increased PPARα expression, and enhanced ACC phosphorylation in liver and WAT....
Source: Journal of Functional Foods - Category: Nutrition Source Type: research
AbstractPurpose of ReviewThe purpose of this update is to summarize the recent advances on the management of cerebral venous thrombosis (CVT).Recent FindingsThere is a trend in declining frequency of CVT patients presenting with focal deficits or coma and a decrease in mortality over time. Anemia and obesity were identified as risk factors for CVT. During pregnancy and puerperium, the higher risk of CVT occurs in the first months post-delivery. With appropriate management, 1/3 of comatose CVT patients can have a full recovery.SummaryThe management of CVT patients includes treatment of associated conditions, anticoagulation...
Source: Current Neurology and Neuroscience Reports - Category: Neuroscience Source Type: research
Publication date: Available online 21 August 2019Source: Anaesthesia &Intensive Care MedicineAuthor(s): Naomi Fleming, Rowena CockerhamAbstractGeneral anaesthesia in obstetrics is reducing with a reported use in only 4% of caesarean sections in 2017–2018. Obstetric surgery is often urgent, requiring effective team communication and a rapid, focused preoperative assessment. Physiological changes of pregnancy increase the incidence of aspiration, desaturation and failed intubation. In addition, the rapidly evolving circumstances add additional stress, impacting on performance. Hypotension from aortocaval compr...
Source: Anaesthesia and intensive care medicine - Category: Anesthesiology Source Type: research
In this study, we provide mechanistic links between FAM134B and adipocyte differentiation. Here, we found that adipocyte-specific FAM134B overexpression mice are obese and have increased white adipose tissue (WAT) mass. Serum tests showed that they developed high glucose level and severe insulin resistance. In addition, they also exhibited enhanced autophagy and reduced mitochondria amount, suggesting the function of FAM134B to promote autophagy in adipocytes. Overexpression of FAM134B in 3 T3-L1 preadipocytes promoted autophagy and differentiation, while the effect could be inhibited after treatment with autophagy ...
Source: Biochimica et Biophysica Acta (BBA) Molecular and Cell Biology of Lipids - Category: Lipidology Source Type: research
We present the results of a social programme facilitating access to physical activities for vulnerable children in Italy. 311 school-age children enrolled in the programme between 2015 and 2017 were assessed for health and lifestyle, anthropometric and nutritional status. Data were compared by origin (Italians vs. immigrants) and then immigrants were split into sub-groups: first- and second-generation. Poor socio-economic status exposed children to a lack of access to health services, and drove imbalanced eating behaviour. 20.8% of children were not registered with the National Health Services (immigrants p 
Source: International Journal of Food Sciences and Nutrition - Category: Nutrition Tags: Int J Food Sci Nutr Source Type: research
This summer, not unlike many families across the country, we were fortunate enough to visit a theme park while celebrating our break from another challenging school year. Being a mother and a pediatrician, I must admit how concerned I was to witness the number of children with obesity attending the park. While we waited in […]Find jobs at  Careers by KevinMD.com.  Search thousands of physician, PA, NP, and CRNA jobs now.  Learn more.
Source: Kevin, M.D. - Medical Weblog - Category: General Medicine Authors: Tags: Physician Obesity Primary Care Source Type: blogs
Conditions:   Overfed;   Obesity Interventions:   Other: Overfeeding diet;   Other: Isocaloric diet Sponsor:   University of Copenhagen Recruiting
Source: ClinicalTrials.gov - Category: Research Source Type: clinical trials
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