47 research outputs found
Measuring <i>KRAS </i>Mutations in Circulating Tumor DNA by Droplet Digital PCR and Next-Generation Sequencing
Measuring total cell-free DNA (cfDNA) or cancer-specific mutations herein has presented as new tools in aiding the treatment of cancer patients. Studies show that total cfDNA bears prognostic value in metastatic colorectal cancer (mCRC) and that measuring cancer-specific mutations could supplement biopsies. However, limited information is available on the performance of different methods. Blood samples from 28 patients with mCRC and known KRAS mutation status were included. cfDNA was extracted and quantified with droplet digital polymerase chain reaction (ddPCR) measuring Beta-2 Microglobulin. KRAS mutation detection was performed using ddPCR (Bio-Rad) and next-generation sequencing (NGS, Ion Torrent PGM). Comparing KRAS mutation status in plasma and tissue revealed concordance rates of 79% and 89% for NGS and ddPCR. Strong correlation between the methods was observed. Most KRAS mutations were also detectable in 10-fold diluted samples using the ddPCR. We find that for detection of KRAS mutations in ctDNA ddPCR was superior to NGS both in analysis success rate and concordance to tissue. We further present results indicating that lower amount of plasma may be used for detection of KRAS mutations in mCRC
Total cell-free DNA, carcinoembryonic antigen, and C-reactive protein for assessment of prognosis in patients with metastatic colorectal cancer
From Ganglion Cell to Photoreceptor Layer: Timeline of Deterioration in a Rat Ischemia/Reperfusion Model
Neuronal damage and impaired vision in different retinal disorders are induced, among other factors, by ischemia/reperfusion (I/R). Since the mechanisms and the progression of ischemic injury are still not completely clarified, a timeline of this retinal degeneration is needed. In this study, we investigated protein and mRNA alterations at 2, 6, 12, and 24 h as well as 3 and 7 days after ischemia to determine the course of an ischemic insult through the whole retina. Moreover, functional analyses were performed at later stages. We detected a significant functional loss of cells in the inner nuclear layer and photoreceptors at 3 and 7 days. Additionally, the thickness of the whole retina was decreased at these points in time, indicating a severe degradation of all retinal layers. Immunohistological and qRT-PCR analyses of retinal ganglion cells (RGCs), glial cells, AII amacrine, cone and rod bipolar as well as cone and rod photoreceptor cells confirmed this first assumption. Our results show that all investigated cell types were damaged by ischemia induction. Especially RGCs, cone bipolar cells, and photoreceptor cones are very sensitive to I/R. These cells were lost shortly after ischemia induction with a progressive course up to 7 days. In addition, Müller cell gliosis was observed over the entire period of time. These results provide evidence, that I/R induces damage of the whole retina at early stages and increases over time. In conclusion, our study could demonstrate the intense impact of an ischemic injury. The ischemic defect spreads across the whole retina right up to the outer layers in the long-term and thus seems to impair the visual perception already during the stimulus processing. In addition, our findings indicate that the cone pathway seems to be particularly affected by this damage
Genome of the Asian Longhorned Beetle (\u3cem\u3eAnoplophora glabripennis\u3c/em\u3e), a Globally Significant Invasive Species, Reveals Key Functional and Evolutionary Innovations at the Beetle-Plant Interface
Background: Relatively little is known about the genomic basis and evolution of wood-feeding in beetles. We undertook genome sequencing and annotation, gene expression assays, studies of plant cell wall degrading enzymes, and other functional and comparative studies of the Asian longhorned beetle, Anoplophora glabripennis, a globally significant invasive species capable of inflicting severe feeding damage on many important tree species. Complementary studies of genes encoding enzymes involved in digestion of woody plant tissues or detoxification of plant allelochemicals were undertaken with the genomes of 14 additional insects, including the newly sequenced emerald ash borer and bull-headed dung beetle. Results: The Asian longhorned beetle genome encodes a uniquely diverse arsenal of enzymes that can degrade the main polysaccharide networks in plant cell walls, detoxify plant allelochemicals, and otherwise facilitate feeding on woody plants. It has the metabolic plasticity needed to feed on diverse plant species, contributing to its highly invasive nature. Large expansions of chemosensory genes involved in the reception of pheromones and plant kairomones are consistent with the complexity of chemical cues it uses to find host plants and mates. Conclusions: Amplification and functional divergence of genes associated with specialized feeding on plants, including genes originally obtained via horizontal gene transfer from fungi and bacteria, contributed to the addition, expansion, and enhancement of the metabolic repertoire of the Asian longhorned beetle, certain other phytophagous beetles, and to a lesser degree, other phytophagous insects. Our results thus begin to establish a genomic basis for the evolutionary success of beetles on plants
New loci for body fat percentage reveal link between adiposity and cardiometabolic disease risk
To increase our understanding of the genetic basis of adiposity and its links to cardiometabolic disease risk, we conducted a genome-wide association meta-analysis of body fat percentage (BF%) in up to 100,716 individuals. Twelve loci reached genome-wide significance (P<5 × 10−8), of which eight were previously associated with increased overall adiposity (BMI, BF%) and four (in or near COBLL1/GRB14, IGF2BP1, PLA2G6, CRTC1) were novel associations with BF%. Seven loci showed a larger effect on BF% than on BMI, suggestive of a primary association with adiposity, while five loci showed larger effects on BMI than on BF%, suggesting association with both fat and lean mass. In particular, the loci more strongly associated with BF% showed distinct cross-phenotype association signatures with a range of cardiometabolic traits revealing new insights in the link between adiposity and disease risk
Polygenic prediction of educational attainment within and between families from genome-wide association analyses in 3 million individuals
We conduct a genome-wide association study (GWAS) of educational attainment (EA) in a sample of ~3 million individuals and identify 3,952 approximately uncorrelated genome-wide-significant single-nucleotide polymorphisms (SNPs). A genome-wide polygenic predictor, or polygenic index (PGI), explains 12-16% of EA variance and contributes to risk prediction for ten diseases. Direct effects (i.e., controlling for parental PGIs) explain roughly half the PGI's magnitude of association with EA and other phenotypes. The correlation between mate-pair PGIs is far too large to be consistent with phenotypic assortment alone, implying additional assortment on PGI-associated factors. In an additional GWAS of dominance deviations from the additive model, we identify no genome-wide-significant SNPs, and a separate X-chromosome additive GWAS identifies 57
Genome of the Asian longhorned beetle (Anoplophora glabripennis), a globally significant invasive species, reveals key functional and evolutionary innovations at the beetle–plant interface
Background
Relatively little is known about the genomic basis and evolution of wood-feeding in beetles. We undertook genome sequencing and annotation, gene expression assays, studies of plant cell wall degrading enzymes, and other functional and comparative studies of the Asian longhorned beetle, Anoplophora glabripennis, a globally significant invasive species capable of inflicting severe feeding damage on many important tree species. Complementary studies of genes encoding enzymes involved in digestion of woody plant tissues or detoxification of plant allelochemicals were undertaken with the genomes of 14 additional insects, including the newly sequenced emerald ash borer and bull-headed dung beetle.
Results
The Asian longhorned beetle genome encodes a uniquely diverse arsenal of enzymes that can degrade the main polysaccharide networks in plant cell walls, detoxify plant allelochemicals, and otherwise facilitate feeding on woody plants. It has the metabolic plasticity needed to feed on diverse plant species, contributing to its highly invasive nature. Large expansions of chemosensory genes involved in the reception of pheromones and plant kairomones are consistent with the complexity of chemical cues it uses to find host plants and mates.
Conclusions
Amplification and functional divergence of genes associated with specialized feeding on plants, including genes originally obtained via horizontal gene transfer from fungi and bacteria, contributed to the addition, expansion, and enhancement of the metabolic repertoire of the Asian longhorned beetle, certain other phytophagous beetles, and to a lesser degree, other phytophagous insects. Our results thus begin to establish a genomic basis for the evolutionary success of beetles on plants
New loci for body fat percentage reveal link between adiposity and cardiometabolic disease risk
To increase our understanding of the genetic basis of adiposity and its links to cardiometabolic disease risk, we conducted a genome-wide association meta-analysis of body fat percentage (BF%) in up to 100,716 individuals. Twelve loci reached genome-wide significance (P <5 x 10(-8)), of which eight were previously associated with increased overall adiposity (BMI, BF%) and four (in or near COBLL1/GRB14, IGF2BP1, PLA2G6, CRTC1) were novel associations with BF%. Seven loci showed a larger effect on BF% than on BMI, suggestive of a primary association with adiposity, while five loci showed larger effects on BMI than on BF%, suggesting association with both fat and lean mass. In particular, the loci more strongly associated with BF% showed distinct cross-phenotype association signatures with a range of cardiometabolic traits revealing new insights in the link between adiposity and disease risk.Peer reviewe
Effect of free fatty acids and triglycerides on the expression of proinflammatory mediators and adhesion molecules in hepatocytes and Kupffer cells (of the rat)
Die Nichtalkoholische Fettlebererkrankung ist eine der häufigsten Lebererkrankungen weltweit. Bisher veröffentlichte Daten aus klinischen sowie tierexperimentellen Studien implizieren, dass die Frühformen dieser Erkrankung, Steatosis sowie Steatohepatitis, auf eine exzessive Exposition von freien Fettsäuren und inflammatorischen Zytokinen zurückzuführen sind. Der exakte Pathomechanismus, der die Progression einer NAFLD zu dem schweren Krankheitsbild einer NASH beschreibt, ist bis auf weiteres noch nicht ausreichend verstanden. Man geht jedoch davon aus, dass unterschiedliche Zellpopulationen in unterschiedlicher Weise in diesen Krankheitsprozess involviert sind. In dieser Arbeit sollte geklärt werden, inwieweit eine Fettexposition eine Rolle in einer möglichen Induktion bzw. Unterhaltung einer Entzündungsreaktion einnimmt. Hierzu wurde gesondert untersucht, wie sich die Genexpression von proinflammatorischen Mediatoren in Hepatozyten- und Kupffer-Zell-Populationen unter der Applikation von Triglyceriden und freien Fettsäuren verändert. Daten dieser Dissertation haben gezeigt, dass die alleinige Applikation von Triglyceriden keine Änderung in der Genexpression von proinflammatorischen Mediatoren sowohl in Hepatozyten wie auch in Kupffer-Zellen der Ratte bewirkt. Auf der anderen Seite können die gewonnenen Daten daraufhin weisen, dass die Fettexposition auch einen leberzellprotektiven Effekt in Hepatozyten aufweisen kann. Unter Entzündungsbedingungen schwächen Hepatozyten scheinbar bei der simultanen Gabe von Triglyceriden und LPS, im Gegensatz zu den Kupffer-Zellen, die die Entzündungsreaktion potenzieren, die eigentliche Entzündungsreaktion tendenziell ab. Dies äußert sich in einer Verminderung der Genexpression von proinflammatorischen Mediatoren. Akkumulieren jedoch Hepatozyten im Vorfeld bereits intrazellulär Triglyceride, so reagieren diese auf einen gesetzten Entzündungsreiz empfindlicher. Im Gegensatz zur Applikation von Triglyceriden konnte beim Einsatz der freien Fettsäure Linolsäure ein zytotoxischer Effekt nachgewiesen werden. Die Daten dieser Arbeit zeigen, dass Linolsäure in einer hohen Konzentration insbesondere Hepatozyten wie auch in einem etwas geringeren Umfang Kupffer-Zellen abtöten. Wie bereits unter Triglyceridexposition detektiert, weisen Hepatozyten auch unter der Applikation von Linolsäure in einer niedrigen Konzentration eher die Tendenz zu einer verminderten Expression von proinflammatorischen Mediatoren auf, so dass man vielmehr von einem protektiven Effekt ausgehen kann. Wie auch unter der Applikation von Triglyceriden zeigen die Daten, dass Kupffer-Zellen unter einer simultanen Gabe von Linolsäure in einer geringen Konzentration und von Endotoxinen eine signifikant verstärkte Induktion der proinflammatorischen Mediatoren im Vergleich zur alleinigen Endotoxinbehandlung aufweisen. Die Daten sprechen dafür, dass Linolsäure die Entzündungsreaktion in Kupffer-Zellen steigert