TY - JOUR
A1 - Enssle, Jörg
A1 - Weylandt, Karsten-Henrich
T1 - Secure and optimized detection of PNPLA3 rs738409 genotype by an improved PCR-restriction fragment length polymorphism method
JF - BioTechniques : the international journal of life science methods
N2 - The PNPLA3 reference single-nucleotide polymorphism rs738409 has been identified as a predisposing factor for nonalcoholic fatty liver disease. A simple method based on PCR and restriction fragment length polymorphism (RFLP) analysis had been published to detect the nonpathogenic allele PNPLA3 rs738409 variant. The presence of the pathogenic variant was deduced by the indigestibility of the corresponding PCR product with BtsCI recognizing the nonpathogenic allele. However, one cannot exclude that an enzymatic reaction does not occur for other, more trivial, reasons. For safe and secure detection of the pathogenic PNPLA3 rs738409, we have further developed the PCR-restriction fragment length polymorphism method by adding a second restriction enzyme digest, clearly identifying the correct PNPLA3 alleles and in particular the pathogenic variant.
METHOD SUMMARY
The method presented here represents an improved genetic diagnosis of the PNPLA3 rs738409 alleles based on conventional and inexpensive molecular biological methods. We used methodology based on PCR and restriction fragment length polymorphisms and clearly identified both described alleles by the use of two restriction enzymes. Digestion of individuals' specific PNPLA3 PCR fragments with both enzymes in independent reactions clearly showed the PNPLA3 rs738409 genotype.
KW - n-3 polyunsaturated fatty acid therapies
KW - nonalcoholic fatty liver
KW - disease
KW - PCR– RFLP
KW - PNPLA3
KW - rs738409
Y1 - 2021
U6 - https://doi.org/10.2144/btn-2020-0163
SN - 0736-6205
SN - 1940-9818
VL - 70
IS - 6
SP - 345
EP - 349
PB - Future Science Ltd.
CY - London
ER -
TY - JOUR
A1 - Li, Chen
A1 - Stoma, Svetlana
A1 - Lotta, Luca A.
A1 - Warner, Sophie
A1 - Albrecht, Eva
A1 - Allione, Alessandra
A1 - Arp, Pascal P.
A1 - Broer, Linda
A1 - Buxton, Jessica L.
A1 - Boeing, Heiner
A1 - Langenberg, Claudia
A1 - Codd, Veryan
T1 - Genome-wide association analysis in humans links nucleotide metabolism to leukocyte telomere length
JF - American Journal of Human Genetics
N2 - Leukocyte telomere length (LTL) is a heritable biomarker of genomic aging. In this study, we perform a genome-wide meta-analysis of LTL by pooling densely genotyped and imputed association results across large-scale European-descent studies including up to 78,592 individuals. We identify 49 genomic regions at a false dicovery rate (FDR) < 0.05 threshold and prioritize genes at 31, with five highlighting nucleotide metabolism as an important regulator of LTL. We report six genome-wide significant loci in or near SENP7, MOB1B, CARMIL1 , PRRC2A, TERF2, and RFWD3, and our results support recently identified PARP1, POT1, ATM, and MPHOSPH6 loci. Phenome-wide analyses in >350,000 UK Biobank participants suggest that genetically shorter telomere length increases the risk of hypothyroidism and decreases the risk of thyroid cancer, lymphoma, and a range of proliferative conditions. Our results replicate previously reported associations with increased risk of coronary artery disease and lower risk for multiple cancer types. Our findings substantially expand current knowledge on genes that regulate LTL and their impact on human health and disease.
KW - Mendelian randomization
KW - risk
KW - variants
KW - disease
KW - cancer
KW - loci
KW - database
KW - genes
KW - heart
KW - gwas
Y1 - 2019
VL - 106
IS - 3
PB - Elsevier
CY - Amsterdam
ER -
TY - JOUR
A1 - Henkel, Janin
A1 - Coleman Mac Gregor of Inneregny, Charles Dominic
A1 - Schraplau, Anne
A1 - Jöhrens, Korinna
A1 - Weiss, Thomas Siegfried
A1 - Jonas, Wenke
A1 - Schürmann, Annette
A1 - Püschel, Gerhard Paul
T1 - Augmented liver inflammation in a microsomal prostaglandin E synthase 1 (mPGES-1)-deficient diet-induced mouse NASH model
JF - Scientific Reports
N2 - In a subset of patients, non-alcoholic fatty liver disease (NAFLD) is complicated by cell death and inflammation resulting in non-alcoholic steatohepatitis (NASH), which may progress to fibrosis and subsequent organ failure. Apart from cytokines, prostaglandins, in particular prostaglandin E-2 (PGE(2)), play a pivotal role during inflammatory processes. Expression of the key enzymes of PGE(2) synthesis, cyclooxygenase 2 and microsomal PGE synthase 1 (mPGES-1), was increased in human NASH livers in comparison to controls and correlated with the NASH activity score. Both enzymes were also induced in NASH-diet-fed wild-type mice, resulting in an increase in hepatic PGE(2) concentration that was completely abrogated in mPGES-1-deficient mice. PGE(2) is known to inhibit TNF-alpha synthesis in macrophages. A strong infiltration of monocyte-derived macrophages was observed in NASH-diet-fed mice, which was accompanied with an increase in hepatic TNF-alpha expression. Due to the impaired PGE(2) production, TNF-alpha expression increased much more in livers of mPGES-1-deficient mice or in the peritoneal macrophages of these mice. The increased levels of TNF-alpha resulted in an enhanced IL-1 beta production, primarily in hepatocytes, and augmented hepatocyte apoptosis. In conclusion, attenuation of PGE(2) production by mPGES-1 ablation enhanced the TNF-alpha-triggered inflammatory response and hepatocyte apoptosis in diet-induced NASH.
KW - suppress VLDL secretion
KW - mice lacking
KW - nonalcoholic steatohepatthis
KW - insulin-resistance
KW - rat hepatocytes
KW - kupffer cells
KW - E-2
KW - disease
KW - expression
KW - accumulation
Y1 - 2018
U6 - https://doi.org/10.1038/s41598-018-34633-y
SN - 2045-2322
IS - 8
SP - 1
EP - 11
PB - Nature Research
CY - London
ER -
TY - JOUR
A1 - Henze, Andrea
A1 - Homann, Thomas
A1 - Rohn, Isabelle
A1 - Aschner, Michael A.
A1 - Link, Christopher D.
A1 - Kleuser, Burkhard
A1 - Schweigert, Florian J.
A1 - Schwerdtle, Tanja
A1 - Bornhorst, Julia
T1 - Caenorhabditis elegans as a model system to study post-translational modifications of human transthyretin
JF - Scientific reports
N2 - The visceral protein transthyretin (TTR) is frequently affected by oxidative post-translational protein modifications (PTPMs) in various diseases. Thus, better insight into structure-function relationships due to oxidative PTPMs of TTR should contribute to the understanding of pathophysiologic mechanisms. While the in vivo analysis of TTR in mammalian models is complex, time- and resource-consuming, transgenic Caenorhabditis elegans expressing hTTR provide an optimal model for the in vivo identification and characterization of drug-mediated oxidative PTPMs of hTTR by means of matrix assisted laser desorption/ionization – time of flight – mass spectrometry (MALDI-TOF-MS). Herein, we demonstrated that hTTR is expressed in all developmental stages of Caenorhabditis elegans, enabling the analysis of hTTR metabolism during the whole life-cycle. The suitability of the applied model was verified by exposing worms to D-penicillamine and menadione. Both drugs induced substantial changes in the oxidative PTPM pattern of hTTR. Additionally, for the first time a covalent binding of both drugs with hTTR was identified and verified by molecular modelling.
KW - n-acetyl-cysteine
KW - s-glutathionylation
KW - force-field
KW - c. elegans
KW - life-span
KW - protein
KW - cells
KW - menadione
KW - disease
KW - binding
Y1 - 2016
U6 - https://doi.org/10.1038/srep37346
SN - 2045-2322
VL - 6
PB - Nature Publishing Group
CY - London
ER -