Nucleosome remodeling complexes comprise several large families of chromatin modifiers that integrate multiple epigenetic control signals to play key roles in cell type-specific transcription regulation. We previously isolated a methyl-binding domain protein 2 (MBD2)-containing nucleosome remodeling and deacetylation (NuRD) complex from primary erythroid cells and showed that MBD2 contributes to DNA methylation-dependent embryonic and fetal β-type globin gene silencing during development in vivo. Here we present structural and biophysical details of the coiledcoil interaction between MBD2 and p66α, a critical component of the MBD2-NuRD complex. We show that enforced expression of the isolated p66α coiled-coil domain relieves MBD2-mediated globin gene silencing and that the expressed peptide interacts only with a subset of components of the MBD2-NuRD complex that does not include native p66α or Mi-2. These results demonstrate the central importance of the coiled-coil interaction and suggest that MBD2-dependent DNA methylation-driven gene silencing can be disrupted by selectively targeting this coiled-coil complex.epigenetics | gene regulation D NA methylation involves the enzymatic addition of a methyl group at the C5 position of symmetrically opposed cytosine bases in a double-stranded cytosine-guanosine sequence (CpG). Regions of increased CpG content (CpG islands) often are found associated with promoters and, when methylated, silence expression of the associated gene (1, 2). Although most CpG islands are largely unmethylated in normal adult tissues, a subset of CpG islands is methylated in specific tissue subtypes, stages of differentiation, and development. Importantly, hypermethylation and silencing of tumor suppressor genes represents a pro-oncogenic change found in a wide range of malignancies (3). These observations have raised interest in DNA methylation as both an important genetic regulatory mechanism and a potential therapeutic target for either re-expression of developmentally silenced genes or reversing tumor suppressor gene silencing in cancer (4, 5).The methyl cytosine binding proteins include a family that specifically recognizes the methylated CpG sequence through an ∼60 amino acid methyl-binding domain (MBD). There are five members of the MBD family in mammals: methyl CpG-binding protein 2 (MeCP2), the first to be identified (6), and MBD1 through MBD4 (7). We and others have isolated and characterized an MBD2-containing nucleosome remodeling and deacetylation (NuRD) complex (referred to as "MBD2-NuRD") that binds methylated DNA and regulates transcription of the associated gene (8-10). The MBD2-NuRD complex comprises at least one homolog of six core proteins: MBD2, retinoblastoma protein-associated protein (RbAp46 or -48) Mi-2(α or β), p66(α or β), histone deacetylase (HDAC1 or 2), and metastasis associated (MTA1 or -2) (Fig. 1A). However, the specific interactions involved in the formation of the MBD2-NuRD complex have not been delineated clearly; information that is key to understanding (i) ...

Psychological stress has been proposed as a major contributor to the progression of cardiovascular disease (CVD). Acute mental stress can activate the sympathetic-adrenal-medullary (SAM) axis, eliciting the release of catecholamines (NE and EPI) resulting in the elevation of heart rate (HR) and blood pressure (BP). Combined stress (psychological and physical) can exacerbate these cardiovascular responses, which may partially contribute to the elevated risk of CVD and increased proportionate mortality risks experienced by some occupations (e.g., firefighting and law enforcement). Studies have supported the benefits of physical activity on physiological and psychological health, including the cardiovascular response to acute stress. Aerobically trained individuals exhibit lower sympathetic nervous system (e.g., HR) reactivity and enhanced cardiovascular efficiency (e.g., lower vascular reactivity and decreased recovery time) in response to physical and/or psychological stress. In addition, resistance training has been demonstrated to attenuate cardiovascular responses and improve mental health. This review will examine stress-induced cardiovascular reactivity and plausible explanations for how exercise training and physical fitness (aerobic and resistance exercise) can attenuate cardiovascular responses to stress. This enhanced functionality may facilitate a reduction in the incidence of stroke and myocardial infarction. Finally, this review will also address the interaction of obesity and physical activity on cardiovascular reactivity and CVD.

The epigenetic code of DNA methylation is interpreted chiefly by methyl cytosine binding domain (MBD) proteins which in turn recruit multiprotein co-repressor complexes. We previously isolated one such complex, MBD2-NuRD, from primary erythroid cells and have shown it contributes to embryonic/fetal β-type globin gene silencing during development. This complex has been implicated in silencing tumor suppressor genes in a variety of human tumor cell types. Here we present structural details of chicken MBD2 bound to a methylated DNA sequence from the ρ-globin promoter to which it binds in vivo and mediates developmental transcriptional silencing in normal erythroid cells. While previous studies have failed to show sequence specificity for MBD2 outside of the symmetric mCpG, we find that this domain binds in a single orientation on the ρ-globin target DNA sequence. Further, we show that the orientation and affinity depends on guanine immediately following the mCpG dinucleotide. Dynamic analyses show that DNA binding stabilizes the central β-sheet, while the N- and C-terminal regions of the protein maintain mobility. Taken together, these data lead to a model in which DNA binding stabilizes the MBD2 structure and that binding orientation and affinity is influenced by the DNA sequence surrounding the central mCpG.

The MBD2-NuRD (Nucleosome Remodeling and Deacetylase) complex is an epigenetic reader of DNA methylation that regulates genes involved in normal development and neoplastic diseases. To delineate the architecture and functional interactions of the MBD2-NuRD complex, we previously solved the structures of MBD2 bound to methylated DNA and a coiled-coil interaction between MBD2 and p66α that recruits the CHD4 nucleosome remodeling protein to the complex. The work presented here identifies novel structural and functional features of a previously uncharacterized domain of MBD2 (MBD2IDR). Biophysical analyses show that the MBD2IDR is an intrinsically disordered region (IDR). However, despite this inherent disorder, MBD2IDR increases the overall binding affinity of MBD2 for methylated DNA. MBD2IDR also recruits the histone deacetylase core components (RbAp48, HDAC2 and MTA2) of NuRD through a critical contact region requiring two contiguous amino acid residues, Arg286 and Leu287. Mutating these residues abrogates interaction of MBD2 with the histone deacetylase core and impairs the ability of MBD2 to repress the methylated tumor suppressor gene PRSS8 in MDA-MB-435 breast cancer cells. These findings expand our knowledge of the multi-dimensional interactions of the MBD2-NuRD complex that govern its function.

Obesity-related oxidative stress, the imbalance between pro-oxidants and antioxidants (e.g., nitric oxide), has been linked to metabolic and cardiovascular disease, including endothelial dysfunction and atherosclerosis. Reactive oxygen species (ROS) are essential for physiological functions including gene expression, cellular growth, infection defense, and modulating endothelial function. However, elevated ROS and/or diminished antioxidant capacity leading to oxidative stress can lead to dysfunction. Physical activity also results in an acute state of oxidative stress. However, it is likely that chronic physical activity provides a stimulus for favorable oxidative adaptations and enhanced physiological performance and physical health, although distinct responses between aerobic and anaerobic activities warrant further investigation. Studies support the benefits of dietary modification as well as exercise interventions in alleviating oxidative stress susceptibility. Since obese individuals tend to demonstrate elevated markers of oxidative stress, the implications for this population are significant. Therefore, in this review our aim is to discuss (i) the role of oxidative stress and inflammation as associated with obesity-related diseases, (ii) the potential concerns and benefits of exercise-mediated oxidative stress, and (iii) the advantageous role of dietary modification, including acute or chronic caloric restriction and vitamin D supplementation.

Image-guided tumor ablation refers to a group of treatment modalities that have emerged during the past 2 decades as important tools in the treatment of a wide range of tumors throughout the body. Although most widely recognized in the treatment of hepatic and renal malignancies, the role of thermal ablation has expanded to include lesions of the lung, breast, prostate, bone, as well as other organs and its clinical applications continue to increase. In the following article, we discuss the major thermal ablation modalities, their respective strengths and weaknesses, potential complications and how to avoid them, as well as possible future applications.

The influence of psychological states on physiological responses during exercise is of considerable importance to individuals for which the efficiency of energy production is critical to occupational performance. Numerous studies have shown that aerobic fitness is associated with enhanced cardiovascular efficiency at rest and that responses to mental stress demonstrate evidence of increased sensitivity (relative increase in HR response) and enhanced efficiency (a decrease in absolute HR). However, the effect of aerobic fitness and its impact on cardiorespiratory (CR) responses to psychological stress during exercise has not been investigated. Therefore, the purpose of this study was three-fold; (1) to examine during exercise, anxiety, effort sense, and CR responses to a mental challenge, (2) to examine anxiety and heart rate (HR) responses from rest to exercise with mental challenge between below average fitness (Low Fit) and well-above average fitness (Hi Fit) individuals (exercising at similar relative intensities), and (3) to examine anxiety, effort sense, and CR responses of Low Fit and Hi Fit individuals to a mental challenge during exercise at a similar relative intensity. Twelve Low Fit and eleven Hi Fit subjects participated in two, 32-minute cycle ergometer rides at 65 % of VO2max. In the mental challenge condition (MCC), subjects rode while participating in mentally challenging tasks (Stroop Color-Word task and mental arithmetic) from min 6 to min 14 of the protocol. In the no mental challenge condition (NMCC), subjects exercised at the same intensity and duration without a stressor. Subjects were counter-balanced between fitness levels and condition. HR, VE, VE/VO2, RR, VO2, RER, effort sense (RPE), and state anxiety (SAI) were assessed at 5, 14, 24, and 30 min. SAI was also assessed at - 5 min before exercise and after 15 min of recovery. In addition, the NASA task load index (NTLX) was used to assess perceived overall workload. SAI increased significantly at 14 min in the MCC. NTLX scores indicated that the MCC was perceived as a greater overall workload. Furthermore, HR, VE, VE/VO2, and RR were significantly elevated during the mental challenge condition at 14 min. The Hi Fit subjects tended to respond to the dual stress of exercise and mental challenge with a relative increase in HR, while absolute HR was similar in both groups. An examination of fitness group differences revealed that SAI and NTLX were similar for Low Fit and Hi Fit subjects when exercising in the MCC, although, Hi Fit subjects demonstrated lower HR responses from 6 min to 14 min. VE, VE/VO2, and RR were similar for Low Fit and Hi Fit subjects. These results suggest that psychological stress during physical activity can exacerbate cardiorespiratory responses and suggests that factors that impact CR adjustment to mental challenge from resting baseline may differ from the factors that impact CR adjustment to mental challenge during exercise. Finally, fitness level attenuates HR and may attenuate additional cardiorespiratory response...

The purpose of this study was to examine the cardiorespiratory (CR) and stress hormone responses to a combined physical and mental stress. Eight participants (VO2(max) = 41.24 +/- 6.20 ml kg(-1) min(-1)) completed two experimental conditions, a treatment condition including a 37 min ride at 60% of VO2(max) with participants responding to a computerized mental challenge dual stress condition (DSC) and a control condition of the same duration and intensity without the mental challenge exercise alone condition (EAC). Significant interactions across time were found for CR responses, with heart rate, ventilation, and respiration rate demonstrating higher increases in the DSC. Additionally, norepinephrine was significantly greater in the DSC at the end of the combined challenge. Furthermore, cortisol area-under-the-curve (AUC) was also significantly elevated during the DSC. These results demonstrate that a mental challenge during exercise can exacerbate the stress response, including the release of hormones that have been linked to negative health consequences (cardiovascular, metabolic, autoimmune illnesses).