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Article ; Online: Reverse Phase Protein Arrays for Compound Profiling.

Moerke, Nathan / Fallahi-Sichani, Mohammad

Current protocols in chemical biology

2016 Volume 8, Issue 3, Page(s) 179–196

Abstract: Reverse phase protein arrays (RPPAs), also called reverse phase lysate arrays (RPLAs), involve immobilizing cell or tissue lysates, in small spots, onto solid supports which are then probed with primary antibodies specific for proteins or post- ... ...

Abstract	Reverse phase protein arrays (RPPAs), also called reverse phase lysate arrays (RPLAs), involve immobilizing cell or tissue lysates, in small spots, onto solid supports which are then probed with primary antibodies specific for proteins or post-translational modifications of interest. RPPA assays are well suited for large-scale, high-throughput measurement of protein and PTM levels in cells and tissues. RPPAs are affordable and highly multiplexable, as a large number of arrays can readily be produced in parallel and then probed separately with distinct primary antibodies. This article describes a procedure for treating cells and preparing cell lysates, as well as a procedure for generating RPPAs using these lysates. A method for probing, imaging, and analyzing RPPAs is also described. These procedures are readily adaptable to a wide range of studies of cell signaling in response to drugs and other perturbations. © 2016 by John Wiley & Sons, Inc.
MeSH term(s)	High-Throughput Screening Assays/methods ; Humans ; Optical Imaging/methods ; Protein Array Analysis/methods ; Signal Transduction
Language	English
Publishing date	2016-09-13
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	2160-4762
ISSN (online)	2160-4762
DOI	10.1002/cpch.9
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Article: Fluorescence Polarization (FP) Assays for Monitoring Peptide-Protein or Nucleic Acid-Protein Binding.

Moerke, Nathan J

Current protocols in chemical biology

2009 Volume 1, Issue 1, Page(s) 1–15

Abstract: The technique of fluorescence polarization (FP) is based on the observation that when a fluorescently labeled molecule is excited by polarized light, it emits light with a degree of polarization that is inversely proportional to the rate of molecular ... ...

Abstract	The technique of fluorescence polarization (FP) is based on the observation that when a fluorescently labeled molecule is excited by polarized light, it emits light with a degree of polarization that is inversely proportional to the rate of molecular rotation. This property of fluorescence can be used to measure the interaction of a small labeled ligand with a larger protein and provides a basis for direct and competition binding assays. FP assays are readily adaptable to a high-throughput format, have been used successfully in screens directed against a wide range of targets, and are particularly valuable in screening for inhibitors of protein-protein and protein-nucleic acid interactions when a small binding epitope can be identified for one of the partners. The protocols in this article describe a general procedure for development of FP assays to monitor binding of such a peptide or oligonucleotide to a protein of interest. Curr. Protoc. Chem Biol. 1:1-15. © 2009 by John Wiley & Sons, Inc.
Language	English
Publishing date	2009-12-01
Publishing country	United States
Document type	Journal Article
ISSN	2160-4762
ISSN	2160-4762
DOI	10.1002/9780470559277.ch090102
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Article ; Online: Discovery of DNL343: A Potent, Selective, and Brain-Penetrant eIF2B Activator Designed for the Treatment of Neurodegenerative Diseases.

Craig, Robert A / De Vicente, Javier / Estrada, Anthony A / Feng, Jianwen A / Lexa, Katrina W / Canet, Mark J / Dowdle, William E / Erickson, Rebecca I / Flores, Brittany N / Haddick, Patrick C G / Kane, Lesley A / Lewcock, Joseph W / Moerke, Nathan J / Poda, Suresh B / Sweeney, Zachary / Takahashi, Ryan H / Tong, Vincent / Wang, Jing / Yulyaningsih, Ernie /

Solanoy, Hilda / Scearce-Levie, Kimberly / Sanchez, Pascal E / Tang, Liwei / Xu, Musheng / Zhang, Rui / Osipov, Maksim

Journal of medicinal chemistry

2024 Volume 67, Issue 7, Page(s) 5758–5782

Abstract: Eukaryotic translation initiation factor 2B (eIF2B) is a key component of the integrated stress response (ISR), which regulates protein synthesis and stress granule formation in response to cellular insult. Modulation of the ISR has been proposed as a ... ...

Abstract	Eukaryotic translation initiation factor 2B (eIF2B) is a key component of the integrated stress response (ISR), which regulates protein synthesis and stress granule formation in response to cellular insult. Modulation of the ISR has been proposed as a therapeutic strategy for treatment of neurodegenerative diseases such as vanishing white matter (VWM) disease and amyotrophic lateral sclerosis (ALS) based on its ability to improve cellular homeostasis and prevent neuronal degeneration. Herein, we report the small-molecule discovery campaign that identified potent, selective, and CNS-penetrant eIF2B activators using both structure- and ligand-based drug design. These discovery efforts culminated in the identification of DNL343, which demonstrated a desirable preclinical drug profile, including a long half-life and high oral bioavailability across preclinical species. DNL343 was progressed into clinical studies and is currently undergoing evaluation in late-stage clinical trials for ALS.
MeSH term(s)	Humans ; Neurodegenerative Diseases/drug therapy ; Neurodegenerative Diseases/metabolism ; Amyotrophic Lateral Sclerosis/drug therapy ; Amyotrophic Lateral Sclerosis/metabolism ; Mutation ; Eukaryotic Initiation Factor-2B/genetics ; Eukaryotic Initiation Factor-2B/metabolism ; Brain/metabolism ; Leukoencephalopathies/metabolism
Chemical Substances	Eukaryotic Initiation Factor-2B
Language	English
Publishing date	2024-03-21
Publishing country	United States
Document type	Journal Article
ZDB-ID	218133-2
ISSN	1520-4804 ; 0022-2623
ISSN (online)	1520-4804
ISSN	0022-2623
DOI	10.1021/acs.jmedchem.3c02422
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Article: Development of in-cell Western assays using far-red fluorophores.

Moerke, Nathan J / Hoffman, Gregory R

Current protocols in chemical biology

2011 Volume 3, Issue 1, Page(s) 39–52

Abstract: The in-cell western (ICW) technique is a cell-based immunoassay method for quantitative measurement of protein expression or phosphorylation levels that can be used for both small molecule and siRNA screening. The method involves growth of cells in ... ...

Abstract	The in-cell western (ICW) technique is a cell-based immunoassay method for quantitative measurement of protein expression or phosphorylation levels that can be used for both small molecule and siRNA screening. The method involves growth of cells in microplates, fixation, permeabilization, and staining with specific antibodies and/or cell labeling dyes. ICW assays take advantage of the properties of near-infrared dyes to achieve higher signal-to-noise ratios than are possible for methods utilizing fluorophores in the visible range of the spectrum, and typically involve measurements using two fluorescent channels: one to measure levels of the target of interest, and one to measure total cell number for normalization. The ICW method is readily adaptable to high-throughput format and has been successfully used with a variety of targets and cell lines. The protocols in this unit describe an ICW procedure for quantitative measurement of rpS6-phosphorylation as an endpoint for monitoring mTORC1 signaling in HeLa cells. This assay can be used for small molecule or siRNA screening, and with modification is adaptable to other cell lines and targets. Curr. Protoc. Chem. Biol. 3:39-52 © 2011 by John Wiley & Sons, Inc.
Language	English
Publishing date	2011-03-01
Publishing country	United States
Document type	Journal Article
ISSN	2160-4762
ISSN	2160-4762
DOI	10.1002/9780470559277.ch100153
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Article ; Online: Assay Development and High-Throughput Screening for Inhibitors of Kaposi's Sarcoma-Associated Herpesvirus N-Terminal Latency-Associated Nuclear Antigen Binding to Nucleosomes.

Beauchemin, Chantal / Moerke, Nathan J / Faloon, Patrick / Kaye, Kenneth M

Journal of biomolecular screening

2014 Volume 19, Issue 6, Page(s) 947–958

Abstract: Kaposi's sarcoma-associated herpesvirus (KSHV) has a causative role in several human malignancies, especially in immunocompromised hosts. KSHV latently infects tumor cells and persists as an extrachromosomal episome (plasmid). KSHV latency-associated ... ...

Abstract	Kaposi's sarcoma-associated herpesvirus (KSHV) has a causative role in several human malignancies, especially in immunocompromised hosts. KSHV latently infects tumor cells and persists as an extrachromosomal episome (plasmid). KSHV latency-associated nuclear antigen (LANA) mediates KSHV episome persistence. LANA binds specific KSHV sequence to replicate viral DNA. In addition, LANA tethers KSHV genomes to mitotic chromosomes to efficiently segregate episomes to daughter nuclei after mitosis. N-terminal LANA (N-LANA) binds histones H2A and H2B to attach to chromosomes. Currently, there are no specific inhibitors of KSHV latent infection. To enable high-throughput screening (HTS) of inhibitors of N-LANA binding to nucleosomes, here we develop, miniaturize, and validate a fluorescence polarization (FP) assay that detects fluorophore-labeled N-LANA peptide binding to nucleosomes. We also miniaturize a counterscreen to identify DNA intercalators that nonspecifically inhibit N-LANA binding to nucleosomes, and also develop an enzyme-linked immunosorbent assay to assess N-LANA binding to nucleosomes in the absence of fluorescence. HTS of libraries containing more than 350,000 compounds identified multiple compounds that inhibited N-LANA binding to nucleosomes. No compounds survived all counterscreens, however. More complex small-molecule libraries will likely be necessary to identify specific inhibitors of N-LANA binding to histones H2A and H2B; these assays should prove useful for future screens.
MeSH term(s)	Animals ; Antigens/chemistry ; Antigens, Viral/chemistry ; Antiviral Agents/chemistry ; Cell Survival ; Chickens ; Drug Design ; Enzyme-Linked Immunosorbent Assay ; Erythrocytes/virology ; Fluorescence Polarization ; Fluorescent Dyes/chemistry ; Glutathione Transferase/metabolism ; HeLa Cells ; Herpesvirus 8, Human/drug effects ; High-Throughput Screening Assays ; Histones/chemistry ; Humans ; Mitosis ; Nuclear Proteins/antagonists & inhibitors ; Nuclear Proteins/chemistry ; Nucleosomes/chemistry ; Plasmids/chemistry ; Protein Domains ; Spectrometry, Fluorescence
Chemical Substances	Antigens ; Antigens, Viral ; Antiviral Agents ; Fluorescent Dyes ; Histones ; Nuclear Proteins ; Nucleosomes ; latency-associated nuclear antigen ; Glutathione Transferase (EC 2.5.1.18)
Language	English
Publishing date	2014-02-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1433680-7
ISSN	1552-454X ; 1087-0571
ISSN (online)	1552-454X
ISSN	1087-0571
DOI	10.1177/1087057114520973
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Article ; Online: Understanding LRRK2 kinase activity in preclinical models and human subjects through quantitative analysis of LRRK2 and pT73 Rab10.

Wang, Xiang / Negrou, Elvira / Maloney, Michael T / Bondar, Vitaliy V / Andrews, Shan V / Montalban, Manuel / Llapashtica, Ceyda / Maciuca, Romeo / Nguyen, Hoang / Solanoy, Hilda / Arguello, Annie / Przybyla, Laralynne / Moerke, Nathan J / Huntwork-Rodriguez, Sarah / Henry, Anastasia G

Scientific reports

2021 Volume 11, Issue 1, Page(s) 12900

Abstract: Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson's disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase activity, ...

Abstract	Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson's disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase activity, supporting the therapeutic potential of LRRK2 kinase inhibitors for PD. To better understand the role of LRRK2 in disease and to support the clinical development of LRRK2 inhibitors, quantitative and high-throughput assays to measure LRRK2 levels and activity are needed. We developed and applied such assays to measure the levels of LRRK2 as well as the phosphorylation of LRRK2 itself or one of its substrates, Rab10 (pT73 Rab10). We observed increased LRRK2 activity in various cellular models of disease, including iPSC-derived microglia, as well as in human subjects carrying the disease-linked variant LRRK2 G2019S. Capitalizing on the high-throughput and sensitive nature of these assays, we detected a significant reduction in LRRK2 activity in subjects carrying missense variants in LRRK2 associated with reduced disease risk. Finally, we optimized these assays to enable analysis of LRRK2 activity following inhibition in human peripheral blood mononuclear cells (PBMCs) and whole blood, demonstrating their potential utility as biomarkers to assess changes in LRRK2 expression and activity in the clinic.
MeSH term(s)	Animals ; Biomarkers ; Enzyme Activation ; Enzyme Assays/methods ; Enzyme Assays/standards ; Gene Expression ; High-Throughput Screening Assays ; Humans ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/genetics ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/metabolism ; Leukocytes, Mononuclear/metabolism ; Mice ; Neuroglia/metabolism ; rab GTP-Binding Proteins/genetics ; rab GTP-Binding Proteins/metabolism
Chemical Substances	Biomarkers ; LRRK2 protein, human (EC 2.7.11.1) ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 (EC 2.7.11.1) ; Lrrk2 protein, mouse (EC 2.7.11.1) ; Rab10 protein, human (EC 3.6.1.-) ; Rab10 protein, mouse (EC 3.6.1.-) ; rab GTP-Binding Proteins (EC 3.6.5.2)
Language	English
Publishing date	2021-06-18
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-91943-4
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Article ; Online: Understanding LRRK2 kinase activity in preclinical models and human subjects through quantitative analysis of LRRK2 and pT73 Rab10

Xiang Wang / Elvira Negrou / Michael T. Maloney / Vitaliy V. Bondar / Shan V. Andrews / Manuel Montalban / Ceyda Llapashtica / Romeo Maciuca / Hoang Nguyen / Hilda Solanoy / Annie Arguello / Laralynne Przybyla / Nathan J. Moerke / Sarah Huntwork-Rodriguez / Anastasia G. Henry

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 17

Abstract: Abstract Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson’s disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase ... ...

Abstract	Abstract Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson’s disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase activity, supporting the therapeutic potential of LRRK2 kinase inhibitors for PD. To better understand the role of LRRK2 in disease and to support the clinical development of LRRK2 inhibitors, quantitative and high-throughput assays to measure LRRK2 levels and activity are needed. We developed and applied such assays to measure the levels of LRRK2 as well as the phosphorylation of LRRK2 itself or one of its substrates, Rab10 (pT73 Rab10). We observed increased LRRK2 activity in various cellular models of disease, including iPSC-derived microglia, as well as in human subjects carrying the disease-linked variant LRRK2 G2019S. Capitalizing on the high-throughput and sensitive nature of these assays, we detected a significant reduction in LRRK2 activity in subjects carrying missense variants in LRRK2 associated with reduced disease risk. Finally, we optimized these assays to enable analysis of LRRK2 activity following inhibition in human peripheral blood mononuclear cells (PBMCs) and whole blood, demonstrating their potential utility as biomarkers to assess changes in LRRK2 expression and activity in the clinic.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2021-06-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
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Article ; Online: Systematic analysis of BRAF(V600E) melanomas reveals a role for JNK/c-Jun pathway in adaptive resistance to drug-induced apoptosis.

Fallahi-Sichani, Mohammad / Moerke, Nathan J / Niepel, Mario / Zhang, Tinghu / Gray, Nathanael S / Sorger, Peter K

Molecular systems biology

2015 Volume 11, Issue 3, Page(s) 797

Abstract: Drugs that inhibit RAF/MEK signaling, such as vemurafenib, elicit profound but often temporary anti-tumor responses in patients with BRAF(V) (600E) melanoma. Adaptive responses to RAF/MEK inhibition occur on a timescale of hours to days, involve ... ...

Abstract	Drugs that inhibit RAF/MEK signaling, such as vemurafenib, elicit profound but often temporary anti-tumor responses in patients with BRAF(V) (600E) melanoma. Adaptive responses to RAF/MEK inhibition occur on a timescale of hours to days, involve homeostatic responses that reactivate MAP kinase signaling and compensatory mitogenic pathways, and attenuate the anti-tumor effects of RAF/MEK inhibitors. We profile adaptive responses across a panel of melanoma cell lines using multiplex biochemical measurement, single-cell assays, and statistical modeling and show that adaptation involves at least six signaling cascades that act to reduce drug potency (IC50) and maximal effect (i.e., Emax ≪ 1). Among these cascades, we identify a role for JNK/c-Jun signaling in vemurafenib adaptation and show that RAF and JNK inhibitors synergize in cell killing. This arises because JNK inhibition prevents a subset of cells in a cycling population from becoming quiescent upon vemurafenib treatment, thereby reducing drug Emax. Our findings demonstrate the breadth and diversity of adaptive responses to RAF/MEK inhibition and a means to identify which steps in a signaling cascade are most predictive of phenotypic response.
MeSH term(s)	Apoptosis ; Cell Line, Tumor ; Cell Survival/drug effects ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; Indoles/pharmacology ; MAP Kinase Signaling System ; Melanoma/drug therapy ; Melanoma/genetics ; Mutation ; Protein Kinase Inhibitors/pharmacology ; Proto-Oncogene Proteins B-raf/genetics ; Sulfonamides/pharmacology
Chemical Substances	Indoles ; Protein Kinase Inhibitors ; Sulfonamides ; vemurafenib (207SMY3FQT) ; BRAF protein, human (EC 2.7.11.1) ; Proto-Oncogene Proteins B-raf (EC 2.7.11.1)
Language	English
Publishing date	2015-03-26
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1744-4292
ISSN (online)	1744-4292
DOI	10.15252/msb.20145877
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Article ; Online: Systematic analysis of BRAFV600E melanomas reveals a role for JNK/c‐Jun pathway in adaptive resistance to drug‐induced apoptosis

Mohammad Fallahi‐Sichani / Nathan J Moerke / Mario Niepel / Tinghu Zhang / Nathanael S Gray / Peter K Sorger

Molecular Systems Biology, Vol 11, Iss 3, Pp n/a-n/a (2015)

2015

Abstract: Abstract Drugs that inhibit RAF/MEK signaling, such as vemurafenib, elicit profound but often temporary anti‐tumor responses in patients with BRAFV600E melanoma. Adaptive responses to RAF/MEK inhibition occur on a timescale of hours to days, involve ... ...

Abstract	Abstract Drugs that inhibit RAF/MEK signaling, such as vemurafenib, elicit profound but often temporary anti‐tumor responses in patients with BRAFV600E melanoma. Adaptive responses to RAF/MEK inhibition occur on a timescale of hours to days, involve homeostatic responses that reactivate MAP kinase signaling and compensatory mitogenic pathways, and attenuate the anti‐tumor effects of RAF/MEK inhibitors. We profile adaptive responses across a panel of melanoma cell lines using multiplex biochemical measurement, single‐cell assays, and statistical modeling and show that adaptation involves at least six signaling cascades that act to reduce drug potency (IC50) and maximal effect (i.e., Emax ≪ 1). Among these cascades, we identify a role for JNK/c‐Jun signaling in vemurafenib adaptation and show that RAF and JNK inhibitors synergize in cell killing. This arises because JNK inhibition prevents a subset of cells in a cycling population from becoming quiescent upon vemurafenib treatment, thereby reducing drug Emax. Our findings demonstrate the breadth and diversity of adaptive responses to RAF/MEK inhibition and a means to identify which steps in a signaling cascade are most predictive of phenotypic response.
Keywords	adaptive responses ; BRAFV600E melanomas ; cell‐to‐cell variability ; RAF and MEK inhibitors ; submaximal drug effect ; Biology (General) ; QH301-705.5 ; Medicine (General) ; R5-920
Subject code	572
Language	English
Publishing date	2015-03-01T00:00:00Z
Publisher	Wiley
Document type	Article ; Online
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Article ; Online: Differential determinants of cancer cell insensitivity to antimitotic drugs discriminated by a one-step cell imaging assay.

Tang, Yangzhong / Xie, Tiao / Florian, Stefan / Moerke, Nathan / Shamu, Caroline / Benes, Cyril / Mitchison, Timothy J

Journal of biomolecular screening

2013 Volume 18, Issue 9, Page(s) 1062–1071

Abstract: Cancer cells can be drug resistant due to genetic variation at multiple steps in the drug response pathway, including drug efflux pumping, target mutation, and blunted apoptotic response. These are not discriminated by conventional cell survival assays. ... ...

Abstract	Cancer cells can be drug resistant due to genetic variation at multiple steps in the drug response pathway, including drug efflux pumping, target mutation, and blunted apoptotic response. These are not discriminated by conventional cell survival assays. Here, we report a rapid and convenient high-content cell-imaging assay that measures multiple physiological changes in cells responding to antimitotic small-molecule drugs. Our one-step, no-wash assay uses three dyes to stain living cells and is much more accurate for scoring weakly adherent mitotic and apoptotic cells than conventional antibody-based assays. We profiled responses of 33 cell lines to 8 antimitotic drugs at multiple concentrations and time points using this assay and deposited our data and assay protocols into a public database (http://lincs.hms.harvard.edu/). Our data discriminated between alternative mechanisms that compromise drug sensitivity to paclitaxel and revealed an unexpected bell-shaped dose-response curve for BI2536, a highly selective inhibitor of Polo-like kinases. Our approach can be generalized, is scalable, and should therefore facilitate identification of molecular biomarkers for mechanisms of drug insensitivity in high-throughput screens and other assays.
MeSH term(s)	Antimitotic Agents/chemistry ; Antimitotic Agents/pharmacology ; Apoptosis/drug effects ; Benzamides/chemistry ; Benzamides/pharmacology ; Cell Cycle Proteins/antagonists & inhibitors ; Cell Cycle Proteins/metabolism ; Cell Line, Tumor ; Cell Survival/drug effects ; Dose-Response Relationship, Drug ; Drug Discovery ; Drug Resistance, Neoplasm/drug effects ; Fluorescent Dyes ; Heterocyclic Compounds, 2-Ring/chemistry ; Heterocyclic Compounds, 2-Ring/pharmacology ; High-Throughput Screening Assays ; Humans ; Molecular Imaging/methods ; Paclitaxel/chemistry ; Paclitaxel/pharmacology ; Protein Serine-Threonine Kinases/antagonists & inhibitors ; Protein Serine-Threonine Kinases/metabolism ; Proto-Oncogene Proteins/antagonists & inhibitors ; Proto-Oncogene Proteins/metabolism ; Pteridines ; Small Molecule Libraries/chemistry ; Small Molecule Libraries/pharmacology ; Structure-Activity Relationship ; Polo-Like Kinase 1
Chemical Substances	Antimitotic Agents ; BI 2536 ; Benzamides ; Cell Cycle Proteins ; Fluorescent Dyes ; Heterocyclic Compounds, 2-Ring ; Proto-Oncogene Proteins ; Pteridines ; Small Molecule Libraries ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; Paclitaxel (P88XT4IS4D)
Language	English
Publishing date	2013-06-20
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1433680-7
ISSN	1552-454X ; 1087-0571
ISSN (online)	1552-454X
ISSN	1087-0571
DOI	10.1177/1087057113493804
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