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Article ; Online: Loss of neuronatin promotes "browning" of primary mouse adipocytes while reducing Glut1-mediated glucose disposal.

Gburcik, Valentina / Cleasby, Mark E / Timmons, James A

American journal of physiology. Endocrinology and metabolism

2013 Volume 304, Issue 8, Page(s) E885–94

Abstract: Failure of white adipose tissue to appropriately store excess metabolic substrate seems to underpin obesity-associated type 2 diabetes. Encouraging "browning" of white adipose has been suggested as a therapeutic strategy to help dispose of excess stored ... ...

Abstract	Failure of white adipose tissue to appropriately store excess metabolic substrate seems to underpin obesity-associated type 2 diabetes. Encouraging "browning" of white adipose has been suggested as a therapeutic strategy to help dispose of excess stored lipid and ameliorate the resulting insulin resistance. Genetic variation at the DNA locus encoding the novel proteolipid neuronatin has been associated with obesity, and we recently observed that neuronatin expression is reduced in subcutaneous adipose tissue from obese humans. Thus, to explore the function of neuronatin further, we used RNAi to silence its expression in murine primary adipocyte cultures and examined the effects on adipocyte phenotype. We found that primary adipocytes express only the longer isoform of neuronatin. Loss of neuronatin led to increased mitochondrial biogenesis, indicated by greater intensity of MitoTracker Green staining. This was accompanied by increased expression of UCP1 and the key genes in mitochondrial oxidative phosphorylation, PGC-1α, Cox8b, and Cox4 in primary subcutaneous white adipocytes, indicative of a "browning" effect. In addition, phosphorylation of AMPK and ACC was increased, suggestive of increased fatty acid utilization. Similar, but less pronounced, effects of neuronatin silencing were also noted in primary brown adipocytes. In contrast, loss of neuronatin caused a reduction in both basal and insulin-stimulated glucose uptake and glycogen synthesis, likely mediated by a reduction in Glut1 protein upon silencing of neuronatin. In contrast, loss of neuronatin had no effect on insulin signaling. In conclusion, neuronatin appears to be a novel regulator of browning and metabolic substrate disposal in white adipocytes.
MeSH term(s)	Adipocytes, White/cytology ; Adipocytes, White/physiology ; Adipogenesis/physiology ; Adipose Tissue, Brown/cytology ; Adipose Tissue, Brown/physiology ; Adult ; Animals ; Blood Glucose/metabolism ; Diabetes Mellitus, Type 2/genetics ; Diabetes Mellitus, Type 2/metabolism ; Diabetes Mellitus, Type 2/physiopathology ; Glucose Transporter Type 1/metabolism ; Glycogen/biosynthesis ; Humans ; Membrane Proteins/genetics ; Membrane Proteins/metabolism ; Mice ; Mice, 129 Strain ; Mice, Knockout ; Middle Aged ; Mitochondria/physiology ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Obesity/genetics ; Obesity/metabolism ; Obesity/physiopathology ; Phenotype ; Primary Cell Culture
Chemical Substances	Blood Glucose ; Glucose Transporter Type 1 ; Membrane Proteins ; NNAT protein, human ; Nerve Tissue Proteins ; Nnat protein, mouse ; SLC2A1 protein, human ; Slc2a1 protein, mouse ; Glycogen (9005-79-2)
Language	English
Publishing date	2013-03-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	603841-4
ISSN	1522-1555 ; 0193-1849
ISSN (online)	1522-1555
ISSN	0193-1849
DOI	10.1152/ajpendo.00463.2012
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Article ; Online: The cell-specific activity of the estrogen receptor alpha may be fine-tuned by phosphorylation-induced structural gymnastics.

Gburcik, Valentina / Picard, Didier

Nuclear receptor signaling

2006 Volume 4, Page(s) e005

Abstract: The estrogen receptor alpha (ERalpha) regulates the transcription of target genes by recruiting coregulator proteins through several domains including the two activation functions AF1 and AF2. The contribution of the N-terminally located AF1 activity is ... ...

Abstract	The estrogen receptor alpha (ERalpha) regulates the transcription of target genes by recruiting coregulator proteins through several domains including the two activation functions AF1 and AF2. The contribution of the N-terminally located AF1 activity is particularly important in differentiated cells, and for ERalpha to integrate inputs from other signaling pathways. However, how the phosphorylation of key residues influences AF1 activity has long remained mysterious, in part because the naturally disordered AF1 domain has resisted a structural characterization. The recent discovery of two coregulators that are specific for a phosphorylated form of AF1 suggests that phosphorylation, possibly in conjunction with the subsequent binding of these coregulators, may enforce a stable structure. The binding of the "pioneer" coregulators might facilitate the subsequent recruitment of yet other coregulators. Different AF1 folds may be enabled by the combinatorial action of posttranslational modifications and coregulator binding thereby fine-tuning ERalpha activities in a cell- and promoter-specific fashion.
Language	English
Publishing date	2006-02-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	2230618-3
ISSN	1550-7629 ; 1550-7629
ISSN (online)	1550-7629
ISSN	1550-7629
DOI	10.1621/nrs.04005
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Article ; Online: An essential role for Tbx15 in the differentiation of brown and "brite" but not white adipocytes.

Gburcik, Valentina / Cawthorn, William P / Nedergaard, Jan / Timmons, James A / Cannon, Barbara

American journal of physiology. Endocrinology and metabolism

2012 Volume 303, Issue 8, Page(s) E1053–60

Abstract: The transcription factor Tbx15 is expressed predominantly in brown adipose tissue and in those white adipose depots that are capable of giving rise to brown-in-white ("brite"/"beige") adipocytes. Therefore, we have investigated a possible role here of ... ...

Abstract	The transcription factor Tbx15 is expressed predominantly in brown adipose tissue and in those white adipose depots that are capable of giving rise to brown-in-white ("brite"/"beige") adipocytes. Therefore, we have investigated a possible role here of Tbx15 in brown and brite adipocyte differentiation in vitro. Adipocyte precursors were isolated from interscapular and axilliary brown adipose tissues, inguinal white ("brite") adipose tissue, and epididymal white adipose tissue in 129/Sv mouse pups and differentiated in culture. Differentiation was enhanced by chronic treatment with the PPARγ agonist rosiglitazone plus the sympathetic neurotransmitter norepinephrine. Using short interfering RNAs (siRNA) directed toward Tbx15 in these primary adipocyte cultures, we decreased Tbx15 expression >90%. This resulted in reduced expression levels of adipogenesis markers (PPARγ, aP2). Importantly, Tbx15 knockdown reduced the expression of brown phenotypic marker genes (PRDM16, PGC-1α, Cox8b/Cox4, UCP1) in brown adipocytes and even more markedly in inguinal white adipocytes. In contrast, Tbx15 knockdown had no effect on white adipocytes originating from a depot that is not brite competent in vivo (epididymal). Therefore, Tbx15 may be essential for the development of the adipogenic and thermogenic programs in adipocytes/adipomyocytes capable of developing brown adipocyte features.
MeSH term(s)	Adipocytes, Brown/physiology ; Adipocytes, Brown/ultrastructure ; Adipocytes, White/physiology ; Adipocytes, White/ultrastructure ; Adipogenesis/physiology ; Animals ; Blotting, Western ; Cell Differentiation/physiology ; DNA Primers ; Genetic Markers ; Hypoglycemic Agents/pharmacology ; Mice ; PPAR gamma/agonists ; Phenotype ; RNA/biosynthesis ; RNA/isolation & purification ; RNA, Small Interfering/genetics ; Real-Time Polymerase Chain Reaction ; Rosiglitazone ; T-Box Domain Proteins/genetics ; T-Box Domain Proteins/physiology ; Thiazolidinediones/pharmacology
Chemical Substances	DNA Primers ; Genetic Markers ; Hypoglycemic Agents ; PPAR gamma ; RNA, Small Interfering ; T-Box Domain Proteins ; TBX15 protein, mouse ; Thiazolidinediones ; Rosiglitazone (05V02F2KDG) ; RNA (63231-63-0)
Language	English
Publishing date	2012-08-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	603841-4
ISSN	1522-1555 ; 0193-1849
ISSN (online)	1522-1555
ISSN	0193-1849
DOI	10.1152/ajpendo.00104.2012
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Article: SPBP is a phosphoserine-specific repressor of estrogen receptor alpha.

Gburcik, Valentina / Bot, Nathalie / Maggiolini, Marcello / Picard, Didier

Molecular and cellular biology

2005 Volume 25, Issue 9, Page(s) 3421–3430

Abstract: Multiple signaling pathways stimulate the activity of estrogen receptor alpha (ERalpha) by direct phosphorylation within its N-terminal activation function 1 (AF1). How phosphorylation affects AF1 activity remains poorly understood. We performed a phage ... ...

Abstract	Multiple signaling pathways stimulate the activity of estrogen receptor alpha (ERalpha) by direct phosphorylation within its N-terminal activation function 1 (AF1). How phosphorylation affects AF1 activity remains poorly understood. We performed a phage display screen for human proteins that are exclusively recruited to the phosphorylated form of AF1 and found the stromelysin-1 platelet-derived growth factor-responsive element-binding protein (SPBP). In a purified system, SPBP bound only the in vitro-phosphorylated form of the ERalpha AF1 or the phosphoserine mimic S118E, and the interaction domain could be mapped to a 42-amino-acid fragment of SPBP. In cells, SPBP preferentially interacted with liganded and phosphorylated ERalpha. Functionally, SPBP behaved as a repressor of activated ERalpha, which extends its previously demonstrated roles as a DNA binding transactivation factor and coactivator of other transcription factors. By targeting the phosphorylated form of AF1, SPBP may contribute to attenuating and fine-tuning ERalpha activity. A functional consequence is that SPBP inhibits the proliferation of ERalpha-dependent but not ERalpha-independent breast cancer cell lines, mirroring a reported negative correlation with the ERalpha status of breast tumors.
MeSH term(s)	Amino Acid Sequence ; Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Cell Line ; Cell Proliferation ; Estrogen Receptor alpha/antagonists & inhibitors ; Estrogen Receptor alpha/genetics ; Estrogen Receptor alpha/metabolism ; Female ; Humans ; Immunoprecipitation ; Molecular Sequence Data ; Mutation/genetics ; Peptide Library ; Phosphorylation ; Phosphoserine/metabolism ; Protein Interaction Mapping ; Protein Structure, Tertiary ; RNA, Small Interfering/genetics ; Repressor Proteins/genetics ; Repressor Proteins/metabolism ; Repressor Proteins/physiology ; Response Elements/genetics ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Transcription Factors/physiology
Chemical Substances	Estrogen Receptor alpha ; Peptide Library ; RNA, Small Interfering ; Repressor Proteins ; TCF20 protein, human ; Transcription Factors ; Phosphoserine (17885-08-4)
Language	English
Publishing date	2005-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	779397-2
ISSN	1098-5549 ; 0270-7306
ISSN (online)	1098-5549
ISSN	0270-7306
DOI	10.1128/MCB.25.9.3421-3430.2005
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Article ; Online: Biomarkers of browning of white adipose tissue and their regulation during exercise- and diet-induced weight loss.

Nakhuda, Asif / Josse, Andrea R / Gburcik, Valentina / Crossland, Hannah / Raymond, Frederic / Metairon, Sylviane / Good, Liam / Atherton, Philip J / Phillips, Stuart M / Timmons, James A

The American journal of clinical nutrition

2016 Volume 104, Issue 3, Page(s) 557–565

Abstract: Background: A hypothesis exists whereby an exercise- or dietary-induced negative energy balance reduces human subcutaneous white adipose tissue (scWAT) mass through the formation of brown-like adipocyte (brite) cells. However, the validity of biomarkers ...

Abstract	Background: A hypothesis exists whereby an exercise- or dietary-induced negative energy balance reduces human subcutaneous white adipose tissue (scWAT) mass through the formation of brown-like adipocyte (brite) cells. However, the validity of biomarkers of brite formation has not been robustly evaluated in humans, and clinical data that link brite formation and weight loss are sparse. Objectives: We used rosiglitazone and primary adipocytes to stringently evaluate a set of biomarkers for brite formation and determined whether the expression of biomarker genes in scWAT could explain the change in body composition in response to exercise training combined with calorie restriction in obese and overweight women (n = 79). Design: Gene expression was derived from exon DNA microarrays and preadipocytes from obesity-resistant and -sensitive mice treated with rosiglitazone to generate candidate brite biomarkers from a microarray. These biomarkers were evaluated against data derived from scWAT RNA from obese and overweight women before and after supervised exercise 5 d/wk for 16 wk combined with modest calorie restriction (∼0.84 MJ/d). Results: Forty percent of commonly used brite gene biomarkers exhibited an exon or strain-specific regulation. No biomarkers were positively related to weight loss in human scWAT. Greater weight loss was significantly associated with less uncoupling protein 1 expression (P = 0.006, R(2) = 0.09). In a follow-up global analysis, there were 161 genes that covaried with weight loss that were linked to greater CCAAT/enhancer binding protein α activity (z = 2.0, P = 6.6 × 10(-7)), liver X receptor α/β agonism (z = 2.1, P = 2.8 × 10(-7)), and inhibition of leptin-like signaling (z = -2.6, P = 3.9 × 10(-5)). Conclusion: We identify a subset of robust RNA biomarkers for brite formation and show that calorie-restriction-mediated weight loss in women dynamically remodels scWAT to take on a more-white rather than a more-brown adipocyte phenotype.
MeSH term(s)	Adipocytes, Beige/cytology ; Adipocytes, Beige/drug effects ; Adipocytes, Beige/metabolism ; Adipocytes, Beige/pathology ; Adipocytes, White/cytology ; Adipocytes, White/drug effects ; Adipocytes, White/metabolism ; Adipocytes, White/pathology ; Adipose Tissue, Beige/metabolism ; Adipose Tissue, Beige/pathology ; Adult ; Animals ; Biomarkers/metabolism ; Cells, Cultured ; Combined Modality Therapy ; Diet, Reducing ; Exercise ; Female ; Gene Expression Profiling ; Gene Expression Regulation/drug effects ; Humans ; Hypoglycemic Agents/pharmacology ; Mice, 129 Strain ; Mice, Inbred C57BL ; Overweight/diet therapy ; Overweight/metabolism ; Overweight/pathology ; Overweight/therapy ; RNA, Messenger/metabolism ; Subcutaneous Fat/metabolism ; Subcutaneous Fat/pathology ; Uncoupling Protein 1/genetics ; Uncoupling Protein 1/metabolism ; Weight Loss
Chemical Substances	Biomarkers ; Hypoglycemic Agents ; RNA, Messenger ; Uncoupling Protein 1
Language	English
Publishing date	2016-08-03
Publishing country	United States
Document type	Controlled Clinical Trial ; Journal Article
ZDB-ID	280048-2
ISSN	1938-3207 ; 0002-9165
ISSN (online)	1938-3207
ISSN	0002-9165
DOI	10.3945/ajcn.116.132563
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Article: Biomarkers of browning of white adipose tissue and their regulation during exercise- and diet-induced weight loss

Nakhuda, Asif / Josse, Andrea R / Gburcik, Valentina / Crossland, Hannah / Raymond, Frederic / Metairon, Sylviane / Good, Liam / Atherton, Philip J / Phillips, Stuart M / Timmons, James A

American journal of clinical nutrition. 2016 Sept. 01, v. 104, no. 3

2016

Abstract	Background: A hypothesis exists whereby an exercise- or dietary-induced negative energy balance reduces human subcutaneous white adipose tissue (scWAT) mass through the formation of brown-like adipocyte (brite) cells. However, the validity of biomarkers of brite formation has not been robustly evaluated in humans, and clinical data that link brite formation and weight loss are sparse. Objectives: We used rosiglitazone and primary adipocytes to stringently evaluate a set of biomarkers for brite formation and determined whether the expression of biomarker genes in scWAT could explain the change in body composition in response to exercise training combined with calorie restriction in obese and overweight women (n = 79). Design: Gene expression was derived from exon DNA microarrays and preadipocytes from obesity-resistant and -sensitive mice treated with rosiglitazone to generate candidate brite biomarkers from a microarray. These biomarkers were evaluated against data derived from scWAT RNA from obese and overweight women before and after supervised exercise 5 d/wk for 16 wk combined with modest calorie restriction (∼0.84 MJ/d). Results: Forty percent of commonly used brite gene biomarkers exhibited an exon or strain-specific regulation. No biomarkers were positively related to weight loss in human scWAT. Greater weight loss was significantly associated with less uncoupling protein 1 expression (P = 0.006, R2 = 0.09). In a follow-up global analysis, there were 161 genes that covaried with weight loss that were linked to greater CCAAT/enhancer binding protein α activity (z = 2.0, P = 6.6 × 10−7), liver X receptor α/β agonism (z = 2.1, P = 2.8 × 10−7), and inhibition of leptin-like signaling (z = −2.6, P = 3.9 × 10−5). Conclusion: We identify a subset of robust RNA biomarkers for brite formation and show that calorie-restriction–mediated weight loss in women dynamically remodels scWAT to take on a more-white rather than a more-brown adipocyte phenotype.
Keywords	DNA microarrays ; RNA ; adipocytes ; binding proteins ; biomarkers ; body composition ; energy balance ; exercise ; exons ; gene expression ; liver ; low calorie diet ; mice ; overweight ; phenotype ; weight loss ; white adipose tissue ; women
Language	English
Dates of publication	2016-0901
Size	p. 557-565.
Publishing place	Oxford University Press
Document type	Article
ZDB-ID	280048-2
ISSN	1938-3207 ; 0002-9165
ISSN (online)	1938-3207
ISSN	0002-9165
DOI	10.3945/ajcn.116.132563
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Gene-chip studies of adipogenesis-regulated microRNAs in mouse primary adipocytes and human obesity.

Keller, Pernille / Gburcik, Valentina / Petrovic, Natasa / Gallagher, Iain J / Nedergaard, Jan / Cannon, Barbara / Timmons, James A

BMC endocrine disorders

2011 Volume 11, Page(s) 7

Abstract: Background: Adipose tissue abundance relies partly on the factors that regulate adipogenesis, i.e. proliferation and differentiation of adipocytes. While components of the transcriptional program that initiates adipogenesis is well-known, the importance ...

Abstract	Background: Adipose tissue abundance relies partly on the factors that regulate adipogenesis, i.e. proliferation and differentiation of adipocytes. While components of the transcriptional program that initiates adipogenesis is well-known, the importance of microRNAs in adipogenesis is less well studied. We thus set out to investigate whether miRNAs would be actively modulated during adipogenesis and obesity. Methods: Several models exist to study adipogenesis in vitro, of which the cell line 3T3-L1 is the most well known, albeit not the most physiologically appropriate. Thus, as an alternative, we produced EXIQON microarray of brown and white primary murine adipocytes (prior to and following differentiation) to yield global profiles of miRNAs. Results: We found 65 miRNAs regulated during in vitro adipogenesis in primary adipocytes. We evaluated the similarity of our responses to those found in non-primary cell models, through literature data-mining. When comparing primary adipocyte profiles, with those of cell lines reported in the literature, we found a high degree of difference in 'adipogenesis' regulated miRNAs suggesting that the model systems may not be accurately representing adipogenesis. The expression of 10 adipogenesis-regulated miRNAs were studied using real-time qPCR and then we selected 5 miRNAs, that showed robust expression, were profiled in subcutaneous adipose tissue obtained from 20 humans with a range of body mass indices (BMI, range = 21-48, and all samples have U133+2 Affymetrix profiles provided). Of the miRNAs tested, mir-21 was robustly expressed in human adipose tissue and positively correlated with BMI (R2 = 0.49, p < 0.001). Conclusion: In conclusion, we provide a preliminary analysis of miRNAs associated with primary cell in vitro adipogenesis and demonstrate that the inflammation-associated miRNA, mir-21 is up-regulated in subcutaneous adipose tissue in human obesity. Further, we provide a novel transcriptomics database of EXIQON and Affymetrix adipocyte profiles to facilitate data mining.
Language	English
Publishing date	2011-03-22
Publishing country	England
Document type	Journal Article
ZDB-ID	2091323-0
ISSN	1472-6823 ; 1472-6823
ISSN (online)	1472-6823
ISSN	1472-6823
DOI	10.1186/1472-6823-11-7
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Article ; Online: Gene-chip studies of adipogenesis-regulated microRNAs in mouse primary adipocytes and human obesity

Gallagher Iain J / Petrovic Natasa / Gburcik Valentina / Keller Pernille / Nedergaard Jan / Cannon Barbara / Timmons James A

BMC Endocrine Disorders, Vol 11, Iss 1, p

2011 Volume 7

Abstract: Abstract Background Adipose tissue abundance relies partly on the factors that regulate adipogenesis, i.e. proliferation and differentiation of adipocytes. While components of the transcriptional program that initiates adipogenesis is well-known, the ... ...

Abstract	Abstract Background Adipose tissue abundance relies partly on the factors that regulate adipogenesis, i.e. proliferation and differentiation of adipocytes. While components of the transcriptional program that initiates adipogenesis is well-known, the importance of microRNAs in adipogenesis is less well studied. We thus set out to investigate whether miRNAs would be actively modulated during adipogenesis and obesity. Methods Several models exist to study adipogenesis in vitro , of which the cell line 3T3-L1 is the most well known, albeit not the most physiologically appropriate. Thus, as an alternative, we produced EXIQON microarray of brown and white primary murine adipocytes (prior to and following differentiation) to yield global profiles of miRNAs. Results We found 65 miRNAs regulated during in vitro adipogenesis in primary adipocytes. We evaluated the similarity of our responses to those found in non-primary cell models, through literature data-mining. When comparing primary adipocyte profiles, with those of cell lines reported in the literature, we found a high degree of difference in 'adipogenesis' regulated miRNAs suggesting that the model systems may not be accurately representing adipogenesis. The expression of 10 adipogenesis-regulated miRNAs were studied using real-time qPCR and then we selected 5 miRNAs, that showed robust expression, were profiled in subcutaneous adipose tissue obtained from 20 humans with a range of body mass indices (BMI, range = 21-48, and all samples have U133+2 Affymetrix profiles provided). Of the miRNAs tested, mir-21 was robustly expressed in human adipose tissue and positively correlated with BMI (R2 = 0.49, p < 0.001). Conclusion In conclusion, we provide a preliminary analysis of miRNAs associated with primary cell in vitro adipogenesis and demonstrate that the inflammation-associated miRNA, mir-21 is up-regulated in subcutaneous adipose tissue in human obesity. Further, we provide a novel transcriptomics database of EXIQON and Affymetrix adipocyte profiles to facilitate data mining.
Keywords	primary white and brown adipocytes ; microRNAs ; microarray ; EXIQON ; Affymetrix ; Adipose tissue: adipocyte ; transcriptome ; Internal medicine ; RC31-1245 ; Medicine ; R ; DOAJ:Internal medicine ; DOAJ:Medicine (General) ; DOAJ:Health Sciences
Subject code	500
Language	English
Publishing date	2011-03-01T00:00:00Z
Publisher	BioMed Central
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Time-series transcriptional profiling yields new perspectives on susceptibility to murine osteoarthritis.

Poulet, Blandine / Ulici, Veronica / Stone, Timothy C / Pead, Matthew / Gburcik, Valentina / Constantinou, Eleni / Palmer, Donald B / Beier, Frank / Timmons, James A / Pitsillides, Andrew A

Arthritis and rheumatism

2012 Volume 64, Issue 10, Page(s) 3256–3266

Abstract: Objective: Chronological age is a powerful epidemiologic risk factor for osteoarthritis (OA), a multifactorial disease that is characterized by articular cartilage (AC) degradation. It is unclear from a molecular perspective how aging interacts with OA ... ...

Abstract	Objective: Chronological age is a powerful epidemiologic risk factor for osteoarthritis (OA), a multifactorial disease that is characterized by articular cartilage (AC) degradation. It is unclear from a molecular perspective how aging interacts with OA to produce this risk to AC integrity. To address this key question, we used in vivo time-course analysis of OA development and murine interstrain variability in natural susceptibility to OA to examine changes in non-OA-prone CBA mice versus OA-prone STR/Ort mice, which develop disease that bears significant histologic resemblance to human OA. Through global transcriptome profiling, we attempted to discover the molecular signature linked with both OA vulnerability and progression. Methods: Affymetrix Mouse Gene 1.0 ST Array profiles were generated from AC samples derived from CBA and STR/Ort mice at 3 different ages, corresponding to the stages prior to, at, and late after the natural onset of OA in the STR/Ort mice. Results: We found that the OA in STR/Ort mice exhibited a molecular phenotype resembling human OA, and we pinpointed a central role of NF-κB signaling and the emergence of an immune-related signature in OA cartilage over time. We discovered that, strikingly, young healthy AC has a highly expressed skeletal muscle gene expression program, which is switched off during maturation, but is intriguingly retained in AC during OA development in STR/Ort mice. Conclusion: This study is the first to show that AC chondrocytes share a high-abundance gene-expression program with skeletal muscle. We show that failure to switch this program off, as well as the restoration of this program, is associated with inappropriate expression of NF-κB signaling pathways, skeletal muscle-related genes, and induction and/or progression of OA.
MeSH term(s)	Animals ; Cartilage, Articular/metabolism ; Cartilage, Articular/pathology ; Chondrocytes/metabolism ; Chondrocytes/pathology ; Gene Expression Profiling ; Genotype ; Mice ; Osteoarthritis/genetics ; Osteoarthritis/metabolism ; Osteoarthritis/pathology ; Phenotype ; Tissue Array Analysis
Language	English
Publishing date	2012-07-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	127294-9
ISSN	1529-0131 ; 0004-3591 ; 2326-5191
ISSN (online)	1529-0131
ISSN	0004-3591 ; 2326-5191
DOI	10.1002/art.34572
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Article: Fungi and animals may share a common ancestor to nuclear receptors.

Phelps, Chris / Gburcik, Valentina / Suslova, Elena / Dudek, Peter / Forafonov, Fedor / Bot, Nathalie / MacLean, Morag / Fagan, Richard J / Picard, Didier

Proceedings of the National Academy of Sciences of the United States of America

2006 Volume 103, Issue 18, Page(s) 7077–7081

Abstract: Nuclear receptors (NRs) are a large family of transcription factors. One hallmark of this family is the ligand-binding domain (LBD), for its primary sequence, structure, and regulatory function. To date, NRs have been found exclusively in animals and ... ...

Abstract	Nuclear receptors (NRs) are a large family of transcription factors. One hallmark of this family is the ligand-binding domain (LBD), for its primary sequence, structure, and regulatory function. To date, NRs have been found exclusively in animals and sponges, which has led to the generally accepted notion that they arose with them. We have overcome the limitations of primary sequence searches by combining sequence profile searches with structural predictions at a genomic scale, and have discovered that the heterodimeric transcription factors Oaf1/Pip2 of the budding yeast Saccharomyces cerevisiae contain putative LBDs resembling those of animal NRs. Although the Oaf1/Pip2 LBDs are embedded in an entirely different architecture, the regulation and function of these transcription factors are strikingly similar to those of the mammalian NR heterodimer peroxisome proliferator-activated receptor alpha/retinoid X receptor (PPAR alpha/RXR). We demonstrate that the induction of Oaf1/Pip2 activity by the fatty acid oleate depends on oleate's direct binding to the Oaf1 LBD. The alteration of two amino acids in the predicted ligand-binding pocket of Oaf1 abolishes both ligand binding and the transcriptional response. Hence, LBDs may have arisen as allosteric switches, for example, to respond to nutritional and metabolic ligands, before the animal and fungal lineages diverged.
MeSH term(s)	Amino Acid Sequence ; Animals ; Computational Biology ; DNA-Binding Proteins ; Evolution, Molecular ; Models, Molecular ; Molecular Sequence Data ; Oleic Acid/metabolism ; Protein Conformation ; Protein Structure, Tertiary ; Receptors, Cytoplasmic and Nuclear/chemistry ; Receptors, Cytoplasmic and Nuclear/genetics ; Receptors, Cytoplasmic and Nuclear/metabolism ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Saccharomyces cerevisiae Proteins/chemistry ; Saccharomyces cerevisiae Proteins/genetics ; Saccharomyces cerevisiae Proteins/metabolism ; Sequence Alignment ; Sequence Analysis, DNA ; Transcription Factors/chemistry ; Transcription Factors/genetics ; Transcription Factors/metabolism
Chemical Substances	DNA-Binding Proteins ; OAF1 protein, S cerevisiae ; PIP2 protein, S cerevisiae ; Receptors, Cytoplasmic and Nuclear ; Saccharomyces cerevisiae Proteins ; Transcription Factors ; Oleic Acid (2UMI9U37CP)
Language	English
Publishing date	2006-04-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.0510080103
Database	MEDical Literature Analysis and Retrieval System OnLINE

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