Investigating the Three Factor Model of Avoidant/Restrictive Food Intake Disorder: A Confirmatory Factor Analysis of the Parent-Report Nine Item ARFID Scale

Avoidant/Restrictive Food Intake Disorder (ARFID) is an eating disorder with three presentations. These are sensory sensitivity, a lack of interest in eating or food, and fear of aversive consequences. The present study seeks to confirm the three-dimensional model of ARFID with the Parent-Report Nine Item ARFID Scale, which serves as a clinical measure of severity of ARFID symptoms. Participants were recruited through a pediatric anxiety clinic and given the Autism Quotient (AQ), Multidimensional Anxiety Scale for Children (MASC), Behavioral Pediatric Feeding Assessment, and the Nine Item ARFID Screen (NIAS). The final sample was one hundred and eighty two (n = 182). The present study sought to find construct validity for the NIAS, convergent validity for candidate ARFID mechanisms and the NIAS, and criterion validity between the NIAS and the BPFAS. Structural Equation Modeling (SEM) was utilized to investigate construct validity of the three-factor ARFID model and the three-factor NIAS model with Confirmatory Factor Analysis (CFA). Correlations and regression analyses were utilized to investigate convergent validity between candidate ARFID mechanisms and the NIAS. Correlations and regression analysis were utilized to investigate criterion validity between the BPFAS and the NIAS as well. Each of the hypotheses were supported both by statistically significant results, as well as reflecting results found within the previous literature as well. The present study is the first to my knowledge to test the validity of the NIAS as a diagnostic instrument for ARFID, and has valuable implications for both future research and advancements in clinical practice

Melting Temperature of MAPK14

Protein kinase domains transfer the γ phosphate of an ATP molecule to a serine, threonine, or tyrosine residue of a protein or peptide substrate1. This phosphorylation can activate or deactivate the protein, meaning that kinases are often important regulators of various cell activities. The active sites of kinase domains are highly conserved as they all bind ATP and have similar functions1. Kinases are very popular drug targets as they play a significant role in cell signaling pathways. Kinase dysregulation has been shown to play a role in many cancers and other diseases, making them a popular target for drugs4. Identifying key differences between different kinase domains could help design highly specific drugs, which would minimize the more severe side effects often seen in other, less specific drugs. MAPK14, an important mitogen activated protein kinase, is involved in the regulation of the cell cycle in response to environmental stress and proinflammatory cytokines2. It is a serine-threonine kinase that has been found to regulate the cell cycle at G0, G1/S, and G2/M transition stages. It can differentially monitor cyclin levels, as well as phosphorylate a number of different tumor suppressor proteins2. MAPK14 can also be activated during normal cellular proliferation and differentiation, as it is a key regulator of hematopoiesis and other important processes3. Experiments performed in CHEM 406 lab failed to determine the melting temperature of MAPK14, as it never melted during the experiments performed. To understand why this may have occurred, circular dichroism (CD) experiments were performed using the standard phosphate buffer, as well as varying concentrations of GuHCl in conjunction with the phosphate buffer. The His tag was also removed, and circular dichroism experiments were performed again, also with almost no success. It is likely that the protein is melting, as a difference in the CD spectra can be seen as the temperature was increased slowly. However, it is likely that the initial experiments failed because the temperature increased too rapidly and melting of the protein could not be detected

Bioactivities of Synthesized Curcumin, Curcuminoids, and Their Metal Complexes

Curcumin is a compound, derived from the Curcuma longa plant, which has shown anticancer activity and relatively low toxicity in humans. It has been a subject of significant interest in recent years as an adjuvant chemotherapy agent. Literature data for curcumin and related curcuminoids have shown discrepancies in the irreported bioactivities, and this has spurred disputes in the scientific community about their purported benefits. A possible explanation for these inconsistencies is variability between curcumin sources in purity and content. A systematic study has been conducted on the biological properties of curcumin from various sources, both naturally sourced and synthesized using neat and solvent-based methods. Bioactivities tested included antioxidative capacity and cell growth inhibition. This study has also been expanded to include curcuminoids containing varied electronic and structural features, and biological properties were also evaluated after coordination to a metal

Measurement of Participation: The Role Checklist Version 3: Satisfaction and Performance

Participation in society is an area of interest to both clinicians and population researchers. Measurement of participation is therefore important, yet differences in definition, in terms of both content and scope, have made general agreement on one instrument tool elusive. What is recognized is the need for a theoretically based tool that captures both the insider and the outsider perspective. The outsider perspective, inclusive of the generally held views of a society, supports the utility for aggregating population data, whereas the insider perspective provides the internally held views of an individual needed for client-centered treatment planning. The Role Checklist Version 3 modifies one of the most commonly used assessment tools in occupational therapy practice, has good preliminary psychometric properties, and is theoretically consistent with both the ICF and the Model of Human Occupation. The Model of Human Occupation is the most widely used theoretical model in occupational therapy. This chapter provides an overview of the theoretical development, empirical testing, and implications for use of this participation measure by occupational therapists along with implications for population researchers

Hydrocarbon seepage in the deep seabed links subsurface and seafloor biospheres

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Chakraborty, A., Ruff, S. E., Dong, X., Ellefson, E. D., Li, C., Brooks, J. M., McBee, J., Bernard, B. B., & Hubert, C. R. J. Hydrocarbon seepage in the deep seabed links subsurface and seafloor biospheres. Proceedings of the National Academy of Sciences of the United States of America, 117(20), (2020): 11029-11037, doi: 10.1073/pnas.2002289117.Marine cold seeps transmit fluids between the subseafloor and seafloor biospheres through upward migration of hydrocarbons that originate in deep sediment layers. It remains unclear how geofluids influence the composition of the seabed microbiome and if they transport deep subsurface life up to the surface. Here we analyzed 172 marine surficial sediments from the deep-water Eastern Gulf of Mexico to assess whether hydrocarbon fluid migration is a mechanism for upward microbial dispersal. While 132 of these sediments contained migrated liquid hydrocarbons, evidence of continuous advective transport of thermogenic alkane gases was observed in 11 sediments. Gas seeps harbored distinct microbial communities featuring bacteria and archaea that are well-known inhabitants of deep biosphere sediments. Specifically, 25 distinct sequence variants within the uncultivated bacterial phyla Atribacteria and Aminicenantes and the archaeal order Thermoprofundales occurred in significantly greater relative sequence abundance along with well-known seep-colonizing members of the bacterial genus Sulfurovum, in the gas-positive sediments. Metabolic predictions guided by metagenome-assembled genomes suggested these organisms are anaerobic heterotrophs capable of nonrespiratory breakdown of organic matter, likely enabling them to inhabit energy-limited deep subseafloor ecosystems. These results point to petroleum geofluids as a vector for the advection-assisted upward dispersal of deep biosphere microbes from subsurface to surface environments, shaping the microbiome of cold seep sediments and providing a general mechanism for the maintenance of microbial diversity in the deep sea.We wish to thank Jody Sandel as well as the crew of R/V GeoExplorer for collection of piston cores, onboard core processing, sample preservation, and shipment. Cynthia Kwan and Oliver Horanszky are thanked for assistance with amplicon library preparation. We also wish to thank Jayne Rattray, Daniel Gittins, and Marc Strous for valuable discussions and suggestions, and Rhonda Clark for research support. Collaborations with Andy Mort from the Geological Survey of Canada, and Richard Hatton from Geoscience Wales are also gratefully acknowledged. This work was financially supported by a Mitacs Elevate Postdoctoral Fellowship awarded to A.C.; an Alberta Innovates-Technology Futures/Eyes High Postdoctoral Fellowship to S.E.R.; and a Natural Sciences and Engineering Research Council Strategic Project Grant, a Genome Canada Genomics Applications Partnership Program grant, a Canada Foundation for Innovation grant (CFI-JELF 33752) for instrumentation, and Campus Alberta Innovates Program Chair funding to C.R.J.H

Proteomic and functional mapping of cardiac NaV1.5 channel phosphorylation sites

Phosphorylation of the voltage-gated Na+ (NaV) channel NaV1.5 regulates cardiac excitability, yet the phosphorylation sites regulating its function and the underlying mechanisms remain largely unknown. Using a systematic, quantitative phosphoproteomic approach, we analyzed NaV1.5 channel complexes purified from nonfailing and failing mouse left ventricles, and we identified 42 phosphorylation sites on NaV1.5. Most sites are clustered, and three of these clusters are highly phosphorylated. Analyses of phosphosilent and phosphomimetic NaV1.5 mutants revealed the roles of three phosphosites in regulating NaV1.5 channel expression and gating. The phosphorylated serines S664 and S667 regulate the voltage dependence of channel activation in a cumulative manner, whereas the nearby S671, the phosphorylation of which is increased in failing hearts, regulates cell surface NaV1.5 expression and peak Na+ current. No additional roles could be assigned to the other clusters of phosphosites. Taken together, our results demonstrate that ventricular NaV1.5 is highly phosphorylated and that the phosphorylation-dependent regulation of NaV1.5 channels is highly complex, site specific, and dynamic

Mechanism of transcription initiation and promoter escape by E. coli RNA polymerase

To investigate roles of the discriminator and open complex (OC) lifetime in transcription initiation by Escherichia coli RNA polymerase (RNAP; α 2 ββ'ωσ 70 ), we compare productive and abortive initiation rates, short RNA distributions, and OC lifetime for the λP R and T7A1 promoters and variants with exchanged discriminators, all with the same transcribed region. The discriminator determines the OC lifetime of these promoters. Permanganate reactivity of thymines reveals that strand backbones in open regions of longlived λP R -discriminator OCs are much more tightly held than for shorter-lived T7A1-discriminator OCs. Initiation from these OCs exhibits two kinetic phases and at least two subpopulations of ternary complexes. Long RNA synthesis (constrained to be single round) occurs only in the initial phase (<10 s), at similar rates for all promoters. Less than half of OCs synthesize a full-length RNA; the majority stall after synthesizing a short RNA. Most abortive cycling occurs in the slower phase (>10 s), when stalled complexes release their short RNA and make another without escaping. In both kinetic phases, significant amounts of 8-nt and 10-nt transcripts are produced by longer-lived, λP R -discriminator OCs, whereas no RNA longer than 7 nt is produced by shorter-lived T7A1-discriminator OCs. These observations and the lack of abortive RNA in initiation from short-lived ribosomal promoter OCs are well described by a quantitative model in which ∼1.0 kcal/mol of scrunching free energy is generated per translocation step of RNA synthesis to overcome OC stability and drive escape. The different length-distributions of abortive RNAs released from OCs with different lifetimes likely play regulatory roles. RNA polymerase | open complex lifetime | transcription initiation | abortive RNA | hybrid length M any facets of transcription initiation by E. coli RNA polymerase (RNAP; α 2 ββ′ωσ 70 ) have been elucidated, but significant questions remain about the mechanism or mechanisms by which initial transcribing complexes (ITC) with a short RNA-DNA hybrid decide to advance and escape from the promoter to enter elongation mode, or, alternately, to stall, release their short RNA, and reinitiate (abortive cycling). For RNAP to escape, its sequencespecific interactions with promoter DNA in the binary open complex (OC) must be overcome. The open regions of promoter DNA in the binary OC are the −10 region (six residues, with specific interactions between σ 2.2 and the nontemplate strand), the discriminator region (typically six to eight residues with no consensus sequence, the upstream end of which interacts with σ 1.2 ), and the transcription start site (TSS, +1) and adjacent residue (+2), which are in the active site of RNAP What drives promoter escape? Escape involves disrupting all the favorable interactions involved in forming and stabilizing the binary OC as well as σ-core interactions. Escape from these interactions is fundamentally driven by the favorable chemical (free) energy change of RNA synthesis, but this energy must be stored in the ITC in each step before escape. Proposed means of energy storage as the length of the RNA-DNA hybrid increases include the stresses introduced by scrunching distortions of the discriminator regions of the open strands in the cleft (2, 5, 6) and by unfavorable interactions of the RNA-DNA hybrid with the hairpin loop of σ 3.2 (7-10). Scrunching of the discriminator region of the template strand is proposed to be most significant for Significance The enzyme RNA polymerase (RNAP) transcribes DNA genetic information into RNA. Regulation of transcription occurs largely in initiation; these regulatory mechanisms must be understood. Lifetimes of transcription-capable RNAP-promoter open complexes (OCs) vary greatly, dictated largely by the DNA discriminator region, but the significance of OC lifetime for regulation was unknown. We observe that a significantly longer RNA:DNA hybrid is synthesized before RNAP escapes from long-lived λP R -promoter OCs as compared with shorter-lived T7A1 promoter OCs. We quantify the free energy needed to overcome OC stability and allow escape from the promoter and elongation of the nascent RNA, and thereby predict escape points for ribosomal (rrnB P1) and lacUV5 promoters. Longer-lived OCs produce longer abortive RNAs, which likely have specific regulatory roles

Mechanism of transcription initiation and promoter escape by E. coli RNA polymerase

To investigate roles of the discriminator and open complex (OC) lifetime in transcription initiation by Escherichia coli RNA polymerase (RNAP; α 2 ββ'ωσ 70 ), we compare productive and abortive initiation rates, short RNA distributions, and OC lifetime for the λP R and T7A1 promoters and variants with exchanged discriminators, all with the same transcribed region. The discriminator determines the OC lifetime of these promoters. Permanganate reactivity of thymines reveals that strand backbones in open regions of longlived λP R -discriminator OCs are much more tightly held than for shorter-lived T7A1-discriminator OCs. Initiation from these OCs exhibits two kinetic phases and at least two subpopulations of ternary complexes. Long RNA synthesis (constrained to be single round) occurs only in the initial phase (<10 s), at similar rates for all promoters. Less than half of OCs synthesize a full-length RNA; the majority stall after synthesizing a short RNA. Most abortive cycling occurs in the slower phase (>10 s), when stalled complexes release their short RNA and make another without escaping. In both kinetic phases, significant amounts of 8-nt and 10-nt transcripts are produced by longer-lived, λP R -discriminator OCs, whereas no RNA longer than 7 nt is produced by shorter-lived T7A1-discriminator OCs. These observations and the lack of abortive RNA in initiation from short-lived ribosomal promoter OCs are well described by a quantitative model in which ∼1.0 kcal/mol of scrunching free energy is generated per translocation step of RNA synthesis to overcome OC stability and drive escape. The different length-distributions of abortive RNAs released from OCs with different lifetimes likely play regulatory roles. RNA polymerase | open complex lifetime | transcription initiation | abortive RNA | hybrid length M any facets of transcription initiation by E. coli RNA polymerase (RNAP; α 2 ββ′ωσ 70 ) have been elucidated, but significant questions remain about the mechanism or mechanisms by which initial transcribing complexes (ITC) with a short RNA-DNA hybrid decide to advance and escape from the promoter to enter elongation mode, or, alternately, to stall, release their short RNA, and reinitiate (abortive cycling). For RNAP to escape, its sequencespecific interactions with promoter DNA in the binary open complex (OC) must be overcome. The open regions of promoter DNA in the binary OC are the −10 region (six residues, with specific interactions between σ 2.2 and the nontemplate strand), the discriminator region (typically six to eight residues with no consensus sequence, the upstream end of which interacts with σ 1.2 ), and the transcription start site (TSS, +1) and adjacent residue (+2), which are in the active site of RNAP What drives promoter escape? Escape involves disrupting all the favorable interactions involved in forming and stabilizing the binary OC as well as σ-core interactions. Escape from these interactions is fundamentally driven by the favorable chemical (free) energy change of RNA synthesis, but this energy must be stored in the ITC in each step before escape. Proposed means of energy storage as the length of the RNA-DNA hybrid increases include the stresses introduced by scrunching distortions of the discriminator regions of the open strands in the cleft (2, 5, 6) and by unfavorable interactions of the RNA-DNA hybrid with the hairpin loop of σ 3.2 (7-10). Scrunching of the discriminator region of the template strand is proposed to be most significant for Significance The enzyme RNA polymerase (RNAP) transcribes DNA genetic information into RNA. Regulation of transcription occurs largely in initiation; these regulatory mechanisms must be understood. Lifetimes of transcription-capable RNAP-promoter open complexes (OCs) vary greatly, dictated largely by the DNA discriminator region, but the significance of OC lifetime for regulation was unknown. We observe that a significantly longer RNA:DNA hybrid is synthesized before RNAP escapes from long-lived λP R -promoter OCs as compared with shorter-lived T7A1 promoter OCs. We quantify the free energy needed to overcome OC stability and allow escape from the promoter and elongation of the nascent RNA, and thereby predict escape points for ribosomal (rrnB P1) and lacUV5 promoters. Longer-lived OCs produce longer abortive RNAs, which likely have specific regulatory roles