Steroidogenic acute regulatory protein (StAR)-related lipid transfer (START) domains were first identified from mammalian proteins that bind lipid/sterol ligands via a hydrophobic pocket. In plants, ...predicted START domains are predominantly found in homeodomain leucine zipper (HD-Zip) transcription factors that are master regulators of cell-type differentiation in development. Here we utilized studies of Arabidopsis in parallel with heterologous expression of START domains in yeast to investigate the hypothesis that START domains are versatile ligand-binding motifs that can modulate transcription factor activity.
Our results show that deletion of the START domain from Arabidopsis Glabra2 (GL2), a representative HD-Zip transcription factor involved in differentiation of the epidermis, results in a complete loss-of-function phenotype, although the protein is correctly localized to the nucleus. Despite low sequence similarly, the mammalian START domain from StAR can functionally replace the HD-Zip-derived START domain. Embedding the START domain within a synthetic transcription factor in yeast, we found that several mammalian START domains from StAR, MLN64 and PCTP stimulated transcription factor activity, as did START domains from two Arabidopsis HD-Zip transcription factors. Mutation of ligand-binding residues within StAR START reduced this activity, consistent with the yeast assay monitoring ligand-binding. The D182L missense mutation in StAR START was shown to affect GL2 transcription factor activity in maintenance of the leaf trichome cell fate. Analysis of in vivo protein-metabolite interactions by mass spectrometry provided direct evidence for analogous lipid-binding activity in mammalian and plant START domains in the yeast system. Structural modeling predicted similar sized ligand-binding cavities of a subset of plant START domains in comparison to mammalian counterparts.
The START domain is required for transcription factor activity in HD-Zip proteins from plants, although it is not strictly necessary for the protein's nuclear localization. START domains from both mammals and plants are modular in that they can bind lipid ligands to regulate transcription factor function in a yeast system. The data provide evidence for an evolutionarily conserved mechanism by which lipid metabolites can orchestrate transcription. We propose a model in which the START domain is used by both plants and mammals to regulate transcription factor activity.
Caspases are known to be involved in animal programmed cell death (PCD). The objective of this thesis was to use gene expression analysis and reverse genetics to determine if Arabidopsis metacaspase ...( AtMC) genes play a role in plant PCD. The majority of AtMC genes were found to be expressed nearly constitutively in various tissues, developmental stages, and under various inductive treatments. Transgenic Arabidopsis plants generated with AtMCpromoter::AtMCgene::GUS fusions showed expression of the reporter gene in leaves, vasculature, trichomes, siliques, anthers, and during embryo development. Preliminary phenotypic characterization of single and double Arabidopsis AtMC loss-of-function mutants suggested that the expression of the AtMC genes are highly functionally redundant. Nevertheless, our results suggest that AtMC1, 2, 4, 6 and 9 may be directly involved in rosette and/or stem development. Although this study does not provide a definitive role of MCs in plant PCD, it lays the foundation for their further in-depth analysis.
Software Review of Analytica 1.2 Cox, Louis Anthony; Paige, Kerrie N.; Popken, Douglas
Human and ecological risk assessment,
19/4/1/, Volume:
5, Issue:
2
Journal Article
Peer reviewed
Analytica is an easy-to-learn, easy-to-use modeling tool that allows modelers to represent what they know through influence diagrams. These diagrams show which model quantities are derived from which ...others and indicate by shape and color the roles that different nodes play in the model, e.g., decision variables, chance variables, outcome variables, deterministic functions, or abstractions of sub-models. A wide variety of built-in probability distributions allow uncertainties about input values to be painlessly specified and propagated through the model via a fast, professional Monte-Carlo simulation engine. Resulting uncertainties and sensitivities about any quantity in the model can be viewed with admirable ease and flexibility by selecting among probability density, cumulative distribution, confidence band, sensitivity analysis, and other displays. Analytica features clever hierarchical model management and navigation features that serious model-builders will appreciate and that novice modelers will learn from as they are led to develop well-structured, well-documented models. Simple continuous (compartmental-flow) and Markov chain dynamic simulation models can be built by paying some detailed attention to arrays and indices, although Analytica does not support true discrete-event simulation. Within its chosen domain-uncertainty propagation through influence diagram models-Analytica is by far the easiest and best tool that we have seen.
Abstract Spinal Muscular Atrophy with Respiratory Distress (SMARD1) is a lethal infantile disease, characterized by the loss of motor neurons leading to muscular atrophy, diaphragmatic paralysis, and ...weakness in the trunk and limbs. Mutations in IGHMBP2, a ubiquitously expressed DNA/RNA helicase, have been shown to cause a wide spectrum of motor neuron disease. Though mutations in IGHMBP2 are mostly associated with SMARD1, milder alleles cause the axonal neuropathy, Charcot-Marie-Tooth disease type 2S (CMT2S), and some null alleles are potentially a risk factor for sudden infant death syndrome (SIDS). Variant heterogeneity studied using an allelic series can be informative in order to create a broad spectrum of models that better exhibit the human variation. We previously identified the nmd2J mouse model of SMARD1, as well as two milder CMT2S mouse models. Here, we used CRISPR-Cas9 genome editing to create three new, more severe Ighmbp2 mouse models of SMARD1, including a null allele, a deletion of C495 (C495del) and a deletion of L362 (L362del). Phenotypic characterization of the IGHMBP2L362del homozygous mutants and IGHMBP2C495del homozygous mutants respectively show a more severe disease presentation than the previous nmd2J model. The IGHMBP2L362del mutants lack a clear denervation in the diaphragm while the IGHMBP2C495del mutants display a neurogenic diaphragmatic phenotype as observed in SMARD1 patients. Characterization of the Ighmbp2-null model indicated neo-natal lethality (median lifespan = 0.5 days). These novel strains expand the spectrum of SMARD1 models to better reflect the clinical continuum observed in the human patients with various IGHMBP2 recessive mutations.
Metabolic networks are interconnected and influence diverse cellular processes. The protein-metabolite interactions that mediate these networks are frequently low affinity and challenging to ...systematically discover. We developed mass spectrometry integrated with equilibrium dialysis for the discovery of allostery systematically (MIDAS) to identify such interactions. Analysis of 33 enzymes from human carbohydrate metabolism identified 830 protein-metabolite interactions, including known regulators, substrates, and products as well as previously unreported interactions. We functionally validated a subset of interactions, including the isoform-specific inhibition of lactate dehydrogenase by long-chain acyl-coenzyme A. Cell treatment with fatty acids caused a loss of pyruvate-lactate interconversion dependent on lactate dehydrogenase isoform expression. These protein-metabolite interactions may contribute to the dynamic, tissue-specific metabolic flexibility that enables growth and survival in an ever-changing nutrient environment.
•CMT, MNDs and other neurodegenerative diseases share genetic contributions.•Many of these diseases appear to have shared molecular mechanisms.•Clinicogenetics can be useful for the diagnosis of many ...neurodegenerative diseases.
Charcot-Marie-Tooth (CMT) disease is a progressive and heterogeneous inherited peripheral neuropathy. A myriad of genetic factors have been identified that contribute to the degeneration of motor and sensory axons in a length-dependent manner. Emerging biological themes underlying disease include defects in axonal trafficking, dysfunction in RNA metabolism and protein homeostasis, as well deficits in the cellular stress response. Moreover, genetic contributions to CMT can have overlap with other neuropathies, motor neuron diseases (MNDs) and neurodegenerative disorders. Recent progress in understanding the molecular biology of CMT and overlapping syndromes aids in the search for necessary therapeutic targets.
Abstract
Charcot–Marie–Tooth disease is an inherited peripheral neuropathy that is clinically and genetically heterogenous. Mutations in IGHMBP2, a ubiquitously expressed DNA/RNA helicase, have been ...shown to cause the infantile motor neuron disease spinal muscular atrophy with respiratory distress type 1 (SMARD1), and, more recently, juvenile-onset Charcot–Marie–Tooth disease type 2S (CMT2S). Using CRISPR-cas9 mutagenesis, we developed the first mouse models of CMT2S p.Glu365del (E365del) and p.Tyr918Cys (Y918C). E365del is the first CMT2S mouse model to be discovered and Y918C is the first human CMT2S allele knock-in model. Phenotypic characterization of the homozygous models found progressive peripheral motor and sensory axonal degeneration. Neuromuscular and locomotor assays indicate that both E365del and Y918C mice have motor deficits, while neurobehavioral characterization of sensory function found that E365del mutants have mechanical allodynia. Analysis of femoral motor and sensory nerves identified axonal degeneration, which does not impact nerve conduction velocities in E365del mice, but it does so in the Y918C model. Based on these results, the E365del mutant mouse, and the human allele knock-in, Y918C, represent mouse models with the hallmark phenotypes of CMT2S, which will be critical for understanding the pathogenic mechanisms of IGHMBP2. These mice will complement existing Ighmbp2 alleles modeling SMARD1 to help understand the complex phenotypic and genotypic heterogeneity that is observed in patients with IGHMBP2 variants.
Abstract
Background
Behavioral weight loss programs often lead to significant short‐term weight loss, but long‐term weight maintenance remains a challenge. Most weight maintenance data come from ...clinical trials, in‐person programs, or general population surveys, but there is a need for better understanding of long‐term weight maintenance in real‐world digital programs.
Methods
This observational survey study examined weight maintenance reported by individuals who had used Noom Weight, a digital commercial behavior change program, and identified factors associated with greater weight maintenance. The cross‐sectional survey was completed by 840 individuals who had lost at least 10% of their body weight using Noom Weight 6–24 months prior.
Results
The study found that 75% of individuals maintained at least 5% weight loss after 1 year, and 49% maintained 10% weight loss. On average, 65% of initial weight loss was maintained after 1 year and 57% after 2 years. Habitual behaviors, such as healthy snacking and exercise, were associated with greater weight maintenance, while demographic factors were not.
Conclusion
This study provides real‐world data on the long‐term weight maintenance achieved using a fully digital behavioral program. The results suggest that Noom Weight is associated with successful weight maintenance in a substantial proportion of users. Future research will use a randomized controlled trial to track weight maintenance after random assignment and at a 2 year follow‐up.
Epidemiologic studies show an increased risk of non‐Hodgkin lymphoma (NHL) in patients with autoimmune disease (AD), due to a combination of shared environmental factors and/or genetic factors, or a ...causative cascade: chronic inflammation/antigen‐stimulation in one disease leads to another. Here we assess shared genetic risk in genome‐wide‐association‐studies (GWAS).
Secondary analysis of GWAS of NHL subtypes (chronic lymphocytic leukemia, diffuse large B‐cell lymphoma, follicular lymphoma, and marginal zone lymphoma) and ADs (rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis). Shared genetic risk was assessed by (a) description of regional genetic of overlap, (b) polygenic risk score (PRS), (c)"diseasome", (d)meta‐analysis.
Descriptive analysis revealed few shared genetic factors between each AD and each NHL subtype. The PRS of ADs were not increased in NHL patients (nor vice versa). In the diseasome, NHLs shared more genetic etiology with ADs than solid cancers (p = .0041). A meta‐analysis (combing AD with NHL) implicated genes of apoptosis and telomere length.
This GWAS‐based analysis four NHL subtypes and three ADs revealed few weakly‐associated shared loci, explaining little total risk. This suggests common genetic variation, as assessed by GWAS in these sample sizes, may not be the primary explanation for the link between these ADs and NHLs.
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