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Article ; Online: Standing with giants: W.J. Whelan, a catalyst for international cooperation via the Miami Winter Symposium and the IUBMB.

Puett, David

2021 Volume 73, Issue 8, Page(s) 1002–1004

MeSH term(s)	Biochemistry ; Congresses as Topic ; Florida ; Humans ; International Cooperation ; Serial Publications
Language	English
Publishing date	2021-06-29
Publishing country	England
Document type	Journal Article ; Personal Narrative ; Portrait
ZDB-ID	1492141-8
ISSN	1521-6551 ; 1521-6543
ISSN (online)	1521-6551
ISSN	1521-6543
DOI	10.1002/iub.2518
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Article: How I became a biochemist.

Puett, David

IUBMB life

2005 Volume 57, Issue 11, Page(s) 769–771

MeSH term(s)	Biochemistry/history ; Georgia ; History, 20th Century ; History, 21st Century
Language	English
Publishing date	2005-11
Publishing country	England
Document type	Biography ; Historical Article ; Journal Article ; Portraits
ZDB-ID	1492141-8
ISSN	1521-6551 ; 1521-6543
ISSN (online)	1521-6551
ISSN	1521-6543
DOI	10.1080/15216540500319176
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Article ; Online: Electron paramagnetic resonance spectroscopy of nitroxide-labeled calmodulin.

Bowman, Paula B / Puett, David

The protein journal

2014 Volume 33, Issue 3, Page(s) 267–277

Abstract: Calmodulin (CaM) is a highly conserved calcium-binding protein consisting of two homologous domains, each of which contains two EF-hands, that is known to bind well over 300 proteins and peptides. In most cases the (Ca(2+))(4-)form of CaM leads to the ... ...

Abstract	Calmodulin (CaM) is a highly conserved calcium-binding protein consisting of two homologous domains, each of which contains two EF-hands, that is known to bind well over 300 proteins and peptides. In most cases the (Ca(2+))(4-)form of CaM leads to the activation of a key regulatory enzyme or protein in a myriad of biological processes. Using the nitroxide spin-labeling reagent, 3-(2-iodoacetamido)-2,2,5,5-tetramethyl-1-pyrrolidinyl oxyl, bovine brain CaM was modified at 2-3 methionines with retention of activity as judged by the activation of cyclic nucleotide phosphodiesterase. X-band electron paramagnetic resonance (EPR) spectroscopy was used to measure the spectral changes upon addition of Ca(2+) to the apo-form of spin-labeled protein. A significant loss of spectral intensity, arising primarily from reductions in the heights of the low, intermediate, and high field peaks, accompanied Ca(2+) binding. The midpoint of the Ca(2+)-mediated transition determined by EPR occurred at a higher Ca(2+) concentration than that measured with circular dichroic spectroscopy and enzyme activation. Recent data have indicated that the transition from the apo-state of CaM to the fully saturated form, [(Ca(2+))(4-)CaM], contains a compact intermediate corresponding to [(Ca(2+))(2-)CaM], and the present results suggest that the spin probes are reporting on Ca(2+) binding to the last two sites in the N-terminal domain, i.e. for the [(Ca(2+))(2)-CaM] → [(Ca(2+))(4-)CaM] transition in which the compact structure becomes more extended. EPR of CaM, spin-labeled at methionines, offers a different approach for studying Ca(2+)-mediated conformational changes and may emerge as a useful technique for monitoring interactions with target proteins.
MeSH term(s)	3',5'-Cyclic-GMP Phosphodiesterases/metabolism ; Animals ; Brain/enzymology ; Brain Chemistry ; Calmodulin/analysis ; Calmodulin/chemistry ; Calmodulin/metabolism ; Cattle ; Electron Spin Resonance Spectroscopy/methods ; Nitrogen Oxides/chemistry ; Spin Labels
Chemical Substances	Calmodulin ; Nitrogen Oxides ; Spin Labels ; 3',5'-Cyclic-GMP Phosphodiesterases (EC 3.1.4.35) ; nitroxyl (GFQ4MMS07W)
Language	English
Publishing date	2014-04-10
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2143071-8
ISSN	1875-8355 ; 1572-3887
ISSN (online)	1875-8355
ISSN	1572-3887
DOI	10.1007/s10930-014-9559-9
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Article: Steroidogenesis and early response gene expression in MA-10 Leydig tumor cells following heterologous receptor down-regulation and cellular desensitization.

Chen, Tsuey-Ming / Czerwiec, Frank S / Puett, David

Biochemistry and biophysics reports

2016 Volume 5, Page(s) 305–312

Abstract: The Leydig tumor cell line, MA-10, expresses the luteinizing hormone receptor, a G protein-coupled receptor that, when activated with luteinizing hormone or chorionic gonadotropin (CG), stimulates cAMP production and subsequent steroidogenesis, notably ... ...

Abstract	The Leydig tumor cell line, MA-10, expresses the luteinizing hormone receptor, a G protein-coupled receptor that, when activated with luteinizing hormone or chorionic gonadotropin (CG), stimulates cAMP production and subsequent steroidogenesis, notably progesterone. These cells also respond to epidermal growth factor (EGF) and phorbol esters with increased steroid biosynthesis. In order to probe the intracellular pathways along with heterologous receptor down-regulation and cellular desensitization, cells were preincubated with EGF or phorbol esters and then challenged with CG, EGF, dibutryl-cyclic AMP, and a phorbol ester. Relative receptor numbers, steroid biosynthesis, and expression of the early response genes, JUNB and c-FOS, were measured. It was found that in all cases but one receptor down-regulation and decreased progesterone production were closely coupled under the conditions used; the exception involved preincubation of the cells with EGF followed by addition of CG where the CG-mediated stimulation of steroidogenesis was considerably lower than the level of receptor down-regulation. In a number of instances JUNB and c-FOS expression paralleled the decreases in receptor number and progesterone production, while in some cases these early response genes were affected little if at all by the changes in receptor number. This finding may indicate that even low levels of activated signaling kinases, e.g. protein kinase A, protein kinase C, or receptor tyrosine kinase, may suffice to yield good expression of JUNB and c-FOS, or it may suggest alternative pathways for regulating expression of these two early response genes.
Language	English
Publishing date	2016-01-06
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2831046-9
ISSN	2405-5808
ISSN	2405-5808
DOI	10.1016/j.bbrep.2016.01.005
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Article ; Online: Accelerometer and GPS Data to Analyze Built Environments and Physical Activity.

Tamura, Kosuke / Wilson, Jeffrey S / Goldfeld, Keith / Puett, Robin C / Klenosky, David B / Harper, William A / Troped, Philip J

Research quarterly for exercise and sport

2019 Volume 90, Issue 3, Page(s) 395–402

Abstract: ... ...

Abstract	Purpose
MeSH term(s)	Accelerometry/instrumentation ; Adult ; Aged ; Environment Design ; Exercise ; Female ; Fitness Trackers ; Geographic Information Systems ; Humans ; Male ; Massachusetts ; Middle Aged ; Population Density ; Residence Characteristics ; Walking ; Young Adult
Language	English
Publishing date	2019-06-14
Publishing country	United States
Document type	Journal Article
ZDB-ID	225654-x
ISSN	2168-3824 ; 0270-1367
ISSN (online)	2168-3824
ISSN	0270-1367
DOI	10.1080/02701367.2019.1609649
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Article ; Online: Steroidogenesis and early response gene expression in MA-10 Leydig tumor cells following heterologous receptor down-regulation and cellular desensitization

Tsuey-Ming Chen / Frank S. Czerwiec / David Puett

Biochemistry and Biophysics Reports, Vol 5, Iss C, Pp 305-

2016 Volume 312

Abstract	The Leydig tumor cell line, MA-10, expresses the luteinizing hormone receptor, a G protein-coupled receptor that, when activated with luteinizing hormone or chorionic gonadotropin (CG), stimulates cAMP production and subsequent steroidogenesis, notably progesterone. These cells also respond to epidermal growth factor (EGF) and phorbol esters with increased steroid biosynthesis. In order to probe the intracellular pathways along with heterologous receptor down-regulation and cellular desensitization, cells were preincubated with EGF or phorbol esters and then challenged with CG, EGF, dibutryl-cyclic AMP, and a phorbol ester. Relative receptor numbers, steroid biosynthesis, and expression of the early response genes, JUNB and c-FOS, were measured. It was found that in all cases but one receptor down-regulation and decreased progesterone production were closely coupled under the conditions used; the exception involved preincubation of the cells with EGF followed by addition of CG where the CG-mediated stimulation of steroidogenesis was considerably lower than the level of receptor down-regulation. In a number of instances JUNB and c-FOS expression paralleled the decreases in receptor number and progesterone production, while in some cases these early response genes were affected little if at all by the changes in receptor number. This finding may indicate that even low levels of activated signaling kinases, e.g. protein kinase A, protein kinase C, or receptor tyrosine kinase, may suffice to yield good expression of JUNB and c-FOS, or it may suggest alternative pathways for regulating expression of these two early response genes.
Keywords	Cellular desensitization ; Early response genes ; Epidermal growth factor and receptor ; Human chorionic gonadotropin and receptor ; MA-10 cells ; Steroidogenesis ; Biology (General) ; QH301-705.5 ; Biochemistry ; QD415-436
Subject code	612
Language	English
Publishing date	2016-03-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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Article ; Online: Microarray-based transcriptome profiling of ovarian cancer cells.

Cui, Juan / Xu, Ying / Puett, David

Methods in molecular biology (Clifton, N.J.)

2013 Volume 1049, Page(s) 119–137

Abstract: Transcriptome profiling is a powerful method for monitoring genes and their expression levels under a variety of conditions. Completion of the human genome and advances in high-throughput gene microarray instrumentation enables one to collect large ... ...

Abstract	Transcriptome profiling is a powerful method for monitoring genes and their expression levels under a variety of conditions. Completion of the human genome and advances in high-throughput gene microarray instrumentation enables one to collect large amounts of data in a relatively short time. The challenge then becomes that of data analysis to identify patterns in expression changes and, from there, to relate the observed changes to functional compartments and pathways in cells, tissues, and organisms. Using cultured human ovarian cancer cells as an experimental model cellular system, we describe approaches that are used in analysis of the transcriptome, focusing on those genes encoding proteins and microRNAs. Coupled with other approaches described herein, one can also use the transcriptome to identify potential serum biomarkers, thus providing direction to what usually is a laborious search for low abundance proteins.
MeSH term(s)	Female ; Gene Expression Profiling/methods ; Gene Expression Regulation, Neoplastic ; Humans ; Molecular Biology/methods ; Ovarian Neoplasms/genetics ; Ovarian Neoplasms/metabolism ; Tissue Array Analysis/methods
Language	English
Publishing date	2013
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-62703-547-7_11
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Article ; Online: Determining the affinity of hormone-receptor interaction.

Puett, David / Angelova, Krassimira

Methods in molecular biology (Clifton, N.J.)

2009 Volume 590, Page(s) 1–20

Abstract: Characterization of the binding of a hormone to its cognate receptor is a cornerstone of many studies in molecular and cellular endocrinology since this event represents the beginning of a specific cellular response, generally from a highly regulated ... ...

Abstract	Characterization of the binding of a hormone to its cognate receptor is a cornerstone of many studies in molecular and cellular endocrinology since this event represents the beginning of a specific cellular response, generally from a highly regulated extracellular messenger. The premise of hormone-receptor interaction follows from the law of mass action describing a reversible second-order reaction, hormone plus receptor, to give a non-covalently associated hormone-receptor complex. From this basic principle, a host of useful experimental parameters are available to the interested investigator. This chapter is focused on development of the experimental and mathematical underpinning of hormone-receptor interaction, with emphasis on a gonadotropin, chorionic gonadotropin (or luteinizing hormone), binding to the luteinizing hormone receptor, a member of the G protein-coupled receptor family. The general concepts and approaches developed herein are, however, valid to most interacting systems.
MeSH term(s)	Animals ; Cell Line ; Chorionic Gonadotropin/metabolism ; DNA, Complementary ; Humans ; Protein Binding ; Rats ; Receptors, LH/genetics ; Receptors, LH/metabolism ; Thermodynamics
Chemical Substances	Chorionic Gonadotropin ; DNA, Complementary ; Receptors, LH
Language	English
Publishing date	2009
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-60327-378-7_1
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Article ; Online: Accelerometer and GPS Analysis of Trail Use and Associations With Physical Activity.

Tamura, Kosuke / Wilson, Jeffrey S / Puett, Robin C / Klenosky, David B / Harper, William A / Troped, Philip J

Journal of physical activity & health

2018 Volume 15, Issue 7, Page(s) 523–530

Abstract: Background: Concurrent use of accelerometers and global positioning system (GPS) data can be used to quantify physical activity (PA) occurring on trails. This study examined associations of trail use with PA and sedentary behavior (SB) and quantified on ...

Abstract	Background: Concurrent use of accelerometers and global positioning system (GPS) data can be used to quantify physical activity (PA) occurring on trails. This study examined associations of trail use with PA and sedentary behavior (SB) and quantified on trail PA using a combination of accelerometer and GPS data. Methods: Adults (N = 142) wore accelerometer and GPS units for 1-4 days. Trail use was defined as a minimum of 2 consecutive minutes occurring on a trail, based on GPS data. We examined associations between trail use and PA and SB. On trail minutes of light-intensity, moderate-intensity, and vigorous-intensity PA, and SB were quantified in 2 ways, using accelerometer counts only and with a combination of GPS speed and accelerometer data. Results: Trail use was positively associated with total PA, moderate-intensity PA, and light-intensity PA (P < .05). On trail vigorous-intensity PA minutes were 346% higher when classified with the combination versus accelerometer only. Light-intensity PA, moderate-intensity PA, and SB minutes were 15%, 91%, and 85% lower with the combination, respectively. Conclusions: Adult trail users accumulated more PA on trail use days than on nontrail use days, indicating the importance of these facilities for supporting regular PA. The combination of GPS and accelerometer data for quantifying on trail activity may be more accurate than accelerometer data alone and is useful for classifying intensity of activities such as bicycling.
MeSH term(s)	Accelerometry ; Adult ; Bicycling/physiology ; Exercise/physiology ; Female ; Geographic Information Systems ; Humans ; Male ; Sedentary Behavior ; Walking/physiology
Language	English
Publishing date	2018-03-26
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1543-5474
ISSN (online)	1543-5474
DOI	10.1123/jpah.2016-0667
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Article ; Online: Shear wave elastography and Afirma™ gene expression classifier in thyroid nodules with indeterminate cytology: a comparison study.

Azizi, Ghobad / Keller, James M / Mayo, Michelle L / Piper, Kelé / Puett, David / Earp, Karly M / Malchoff, Carl D

Endocrine

2018 Volume 59, Issue 3, Page(s) 573–584

Abstract: Purpose: To compare shear wave elastography (SWE) and Afirma™ gene expression classifier (GEC) for diagnosis of malignancy in thyroid nodules (TNs) with Bethesda Classification (BC) III or IV indeterminate cytology.: Methods: This preliminary single- ... ...

Abstract	Purpose: To compare shear wave elastography (SWE) and Afirma™ gene expression classifier (GEC) for diagnosis of malignancy in thyroid nodules (TNs) with Bethesda Classification (BC) III or IV indeterminate cytology. Methods: This preliminary single-center prospective study was approved by the Institutional Review Board. We evaluated 151 consented patients with 151 indeterminate TNs (123 BC III, 28 BC IV) on fine-needle aspiration biopsy (FNAB). B-mode ultrasound, vascularity, and SWE were performed prior to FNAB. TN stiffness was measured as shear wave velocity (SWV) in meters per second (m/s). The stiffest area of the TN was selected for SWV measurement. GEC testing was performed with a second FNAB. Surgery was recommended for GEC-suspicious TNs, or GEC-benign TNs with two or more worrisome B-mode US features. Results: Surgical pathology confirmed 31 malignant TNs. Among the GEC-suspicious group, 28 of 59 TNs were malignant. The SWV value of ≥3.59 m/s was the best cut-off for malignancy risk based on the receiver operating curve (ROC). Twenty-six malignant TNs had SWV ≥ 3.59 m/s. The sensitivity and specificity for SWV ≥ 3.59 m/s were 83.9 and 79.2%, respectively. Positive predictive value (PPV) was 51.0% and negative predictive value (NPV) was 95.0%. For the GEC-suspicious group, sensitivity, specificity, PPV, and NPV were 90.3, 74.2, 47.5, and 96.7%, respectively. In multivariate analysis, SWV and GEC-suspicious were significant predictors of malignancy, but B-mode features and vascularity were not. Conclusion: This preliminary study indicates that SWE and GEC are independent predictors of malignancy in TNs with BC III or IV.
MeSH term(s)	Adult ; Aged ; Biopsy, Fine-Needle ; Elasticity Imaging Techniques/methods ; Female ; Gene Expression Profiling ; Humans ; Male ; Middle Aged ; Prospective Studies ; Sensitivity and Specificity ; Thyroid Gland/diagnostic imaging ; Thyroid Gland/pathology ; Thyroid Nodule/diagnostic imaging ; Thyroid Nodule/genetics ; Thyroid Nodule/pathology
Language	English
Publishing date	2018-01-19
Publishing country	United States
Document type	Comparative Study ; Journal Article
ZDB-ID	1194484-5
ISSN	1559-0100 ; 1355-008X ; 0969-711X
ISSN (online)	1559-0100
ISSN	1355-008X ; 0969-711X
DOI	10.1007/s12020-017-1509-9
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