94 research outputs found
Weighted Morrey Estimates for Multilinear Fourier Multiplier Operators
The multilinear Fourier multipliers and their commutators with Sobolev regularity are studied. The purpose of this paper is to establish that these operators are bounded on certain product Morrey spaces Lp,k(ān). Based on the boundedness of these operators from Lp1(Ļ1)ĆāÆĆLpm(Ļm) to Lp(āj=1māĻp/pj), we obtained that they are also bounded from Lp1,k(Ļ1)ĆāÆĆLpm,k(Ļm) to Lp,k(āj=1māĻp/pj), with 0<k<1, 1<pj<ā, 1/p=1/p1+āÆ+1/pm, and ĻjāApj,j=1,ā¦,m
Beta-arrestin inhibits CAMKKbeta-dependent AMPK activation downstream of protease-activated-receptor-2
<p>Abstract</p> <p>Background</p> <p>Proteinase-activated-receptor-2 (PAR<sub>2</sub>) is a seven transmembrane receptor that can activate two separate signaling arms: one through GĪ±q and Ca<sup>2+ </sup>mobilization, and a second through recruitment of Ī²-arrestin scaffolds. In some cases downstream targets of the GĪ±q/Ca<sup>2+ </sup>signaling arm are directly inhibited by Ī²-arrestins, while in other cases the two pathways are synergistic; thus Ī²-arrestins act as molecular switches capable of modifying the signal generated by the receptor.</p> <p>Results</p> <p>Here we demonstrate that PAR<sub>2 </sub>can activate adenosine monophosphate-activated protein kinase (AMPK), a key regulator of cellular energy balance, through Ca<sup>2+</sup>-dependent Kinase Kinase Ī² (CAMKKĪ²), while inhibiting AMPK through interaction with Ī²-arrestins. The ultimate outcome of PAR<sub>2 </sub>activation depended on the cell type studied; in cultured fibroblasts with low endogenous Ī²-arrestins, PAR<sub>2 </sub>activated AMPK; however, in primary fat and liver, PAR<sub>2 </sub>only activated AMPK in Ī²-arrestin-2<sup>-/- </sup>mice. Ī²-arrestin-2 could be co-immunoprecipitated with AMPK and CAMKKĪ² under baseline conditions from both cultured fibroblasts and primary fat, and its association with both proteins was increased by PAR<sub>2 </sub>activation. Addition of recombinant Ī²-arrestin-2 to in vitro kinase assays directly inhibited phosphorylation of AMPK by CAMKKĪ² on Thr172.</p> <p>Conclusions</p> <p>Studies have shown that decreased AMPK activity is associated with obesity and Type II Diabetes, while AMPK activity is increased with metabolically favorable conditions and cholesterol lowering drugs. These results suggest a role for Ī²-arrestin in the inhibition of AMPK signaling, raising the possibility that Ī²-arrestin-dependent PAR<sub>2 </sub>signaling may act as a molecular switch turning a positive signal to AMPK into an inhibitory one.</p
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Low-Level Saturated Fatty Acid Palmitate Benefits Liver Cells by Boosting Mitochondrial Metabolism via CDK1-SIRT3-CPT2 Cascade.
Saturated fatty acids (SFAs) (the "bad" fat), especially palmitate (PA), in the human diet are blamed for potential health risks such as obesity and cancer because of SFA-induced lipotoxicity. However, epidemiological results demonstrate a latent benefit of SFAs, and it remains elusive whether a certain low level of SFAs is physiologically essential for maintaining cell metabolic hemostasis. Here, we demonstrate that although high-level PA (HPA) indeed induces lipotoxic effects in liver cells, low-level PA (LPA) increases mitochondrial functions and alleviates the injuries induced by HPA or hepatoxic agent carbon tetrachloride (CCl4). LPA treatment in mice enhanced liver mitochondrial activity and reduced CCl4 hepatotoxicity with improved blood levels of aspartate aminotransferase (AST), alanine transaminase (ALT), and mitochondrial aspartate transaminase (m-AST). LPA-mediated mitochondrial homeostasis is regulated by CDK1-mediated SIRT3 phosphorylation, which in turn deacetylates and dimerizes CPT2 to enhance fatty acid oxidation. Thus, an advantageous effect is suggested by the consumption of LPA that augments mitochondrial metabolic homeostasis via CDK1-SIRT3-CPT2 cascade
Kinetics of deamination and Cu(II)/H2O2/Ascorbate-induced formation of 5-methylcytosine glycol at CpG sites in duplex DNA
Mutation in p53 tumor suppressor gene is a hallmark of human cancers. Six major mutational hotspots in p53 contain methylated CpG (mCpG) sites, and C āT transition is the most common mutation at these sites. It was hypothesized that the formation of 5-methylcytosine glycol induced by reactive oxygen species, its spontaneous deamination to thymine glycol and the miscoding property of the latter may account, in part, for the ubiquitous C āT mutation at CpG site. Here, we assessed the kinetics of deamination for two diastereomers of 5-methylcytosine glycol in duplex DNA. Our results revealed that the half-lives for the deamination of the (5S,6S) and (5R,6R) diastereomers of 5-methylcytosine glycol in duplex DNA at 37Ā°C were 37.4 Ā± 1.6 and 27.4 Ā± 1.0 h, respectively. The deamination rates were only slightly lower than those for the two diastereomers in mononucleosides. Next, we assessed the formation of 5-methyl-2ā²-deoxycytidine glycol in the form of its deaminated product, namely, thymidine glycol (Tg), in methyl-CpG-bearing duplex DNA treated with Cu(II)/H2O2/ascorbate. LC-MS/MS quantification results showed that the yield of Tg is similar as that of 5-(hydroxymethyl)-2ā²-deoxycytidine. Together, our data support that the formation and deamination of 5-methylcytosine glycol may contribute significantly to the C āT transition mutation at mCpG dinucleotide site
Formation and genotoxicity of a guanineācytosine intrastrand cross-link lesion in vivo
Reactive oxygen species (ROS) can be induced by both endogenous and exogenous processes, and they can damage biological molecules including nucleic acids. Exposure of isolated DNA to X/Ī³-rays and Fenton reagents was shown to lead to the formation of intrastrand cross-link lesions where the neighboring nucleobases in the same DNA strand are covalently bonded. By employing HPLC coupled with tandem mass spectrometry (LC-MS/MS) with the isotope dilution method, we assessed quantitatively the formation of a guanineācytosine (G[8-5]C) intrastrand cross-link lesion in HeLa-S3 cells upon exposure to Ī³-rays. The yield of the G[8-5]C cross-link was 0.037 lesions per 109 nucleosides per Gy, which was ā¼300 times lower than that of 5-formyl-2ā²-deoxyuridine (0.011 lesions per 106 nucleosides per Gy) under identical exposure conditions. We further constructed a single-stranded M13 genome harboring a site-specifically incorporated G[8-5]C lesion and developed a novel mass spectrometry-based method for interrogating the products emanating from the replication of the genome in Escherichia coli cells. The results demonstrated that G[8-5]C blocked considerably DNA replication as represented by a 20% bypass efficiency, and the lesion was significantly mutagenic in vivo, which included a 8.7% GāT and a 1.2% GāC transversion mutations. DNA replication in E. coli hosts deficient in SOS-induced polymerases revealed that polymerase V was responsible for the error-prone translesion synthesis in vivo
Growth Inhibition and Apoptosis Induced by Osthole, A Natural Coumarin, in Hepatocellular Carcinoma
BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most commonly diagnosed tumors worldwide and is known to be resistant to conventional chemotherapy. New therapeutic strategies are urgently needed for treating HCC. Osthole, a natural coumarin derivative, has been shown to have anti-tumor activity. However, the effects of osthole on HCC have not yet been reported. METHODS AND FINDINGS: HCC cell lines were treated with osthole at various concentrations for 24, 48 and 72 hours. The proliferations of the HCC cells were measured by MTT assays. Cell cycle distribution and apoptosis were determined by flow cytometry. HCC tumor models were established in mice by subcutaneously injection of SMMC-7721 or Hepa1-6 cells and the effect of osthole on tumor growths in vivo and the drug toxicity were studied. NF-ĪŗB activity after osthole treatment was determined by electrophoretic mobility shift assays and the expression of caspase-3 was measured by western blotting. The expression levels of other apoptosis-related genes were also determined by real-time PCR (PCR array) assays. Osthole displayed a dose- and time-dependent inhibition of the HCC cell proliferations in vitro. It also induced apoptosis and caused cell accumulation in G2 phase. Osthole could significantly suppress HCC tumor growth in vivo with no toxicity at the dose we used. NF-ĪŗB activity was significantly suppressed by osthole at the dose- and time-dependent manner. The cleaved caspase-3 was also increased by osthole treatment. The expression levels of some apoptosis-related genes that belong to TNF ligand family, TNF receptor family, Bcl-2 family, caspase family, TRAF family, death domain family, CIDE domain and death effector domain family and CARD family were all increased with osthole treatment. CONCLUSION: Osthole could significantly inhibit HCC growth in vitro and in vivo through cell cycle arrest and inducing apoptosis by suppressing NF-ĪŗB activity and promoting the expressions of apoptosis-related genes
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Quantitative Assessment of Arsenite-Induced Perturbation of Ubiquitinated Proteome.
Arsenic contamination in food and groundwater constitutes a public health concern for more than 200 million people worldwide. Individuals chronically exposed to arsenic through drinking and ingestion exhibit a higher risk of developing cancers and cardiovascular diseases. Nevertheless, the underlying mechanisms of arsenic toxicity are not fully understood. Arsenite is known to bind to and deactivate RING finger E3 ubiquitin ligases; thus, we reason that a systematic interrogation about how arsenite exposure modulates global protein ubiquitination may reveal novel molecular targets for arsenic toxicity. By employing liquid chromatography-tandem mass spectrometry, in combination with stable isotope labeling by amino acids in cell culture (SILAC) and immunoprecipitation of di-glycine-conjugated lysine-containing tryptic peptides, we assessed the alterations in protein ubiquitination in GM00637 human skin fibroblast cells upon arsenite exposure at the entire proteome level. We observed that arsenite exposure led to altered ubiquitination of many proteins, where the alterations in a large majority of ubiquitination events are negatively correlated with changes in expression of the corresponding proteins, suggesting their modulation by the ubiquitin-proteasomal pathway. Moreover, we observed that arsenite exposure confers diminished ubiquitination of a rate-limiting enzyme in cholesterol biosynthesis, HMGCR, at Lys248. We also revealed that TRC8 is the major E3 ubiquitin ligase for HMGCR ubiquitination in HEK293T cells, and the arsenite-induced diminution of HMGCR ubiquitination is abrogated upon genetic depletion of TRC8. In summary, we systematically characterized arsenite-induced perturbations in a ubiquitinated proteome in human cells and found that the arsenite-elicited attenuation of HMGCR ubiquitination in HEK293T cells involves TRC8
Multilinear Singular Integrals and their Commutators with Nonsmooth Kernels on Weighted Morrey Spaces
Some multilinear maximal functions and the generalized CalderĆ³n-Zygmund operators and their commutators with nonsmooth kernels are studied. The purpose of this paper is to establish that these operators are bounded on certain product Morrey spaces Lp,k(ān). Based on the boundedness of these operators from Lp1(Ļi)ĆāÆĆLpm(Ļm) to Lp(āj=1māĻjp/pj), we obtained that they are also bounded from Lp1,k(Ļi)ĆāÆĆLpm,k(Ļm) to Lp,k(āj=1māĻjp/pj) with 0<k<1, 1<pj<ā, 1/p=1/p1,ā¦,pm and ĻjāApj, j=1,ā¦,m
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