Salt Induces Adipogenesis/Lipogenesis and Inflammatory Adipocytokines Secretion in Adipocytes

It is well known that high salt intake is associated with cardiovascular diseases including hypertension. However, the research on the mechanism of obesity due to high salt intake is rare. To evaluate the roles of salt on obesity prevalence, the gene expression of adipogenesis/lipogenesis and adipocytokines secretion according to adipocyte dysfunction were investigated in salt-loading adipocytes. High salt dose-dependently increased the expression of adipogenic/lipogenic genes, such as PPAR-γ, C/EBPα, SREBP1c, ACC, FAS, and aP2, but decreased the gene of lipolysis like AMPK, ultimately resulting in fat accumulation. With SIK-2 and Na+/K+-ATPase activation, salt increased the metabolites involved in the renin-angiotensin-aldosterone system (RAAS) such as ADD1, CYP11β2, and MCR. Increasing insulin dependent insulin receptor substrate (IRS)-signaling, resulting in the insulin resistance, mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and Akt-mTOR were activated but AMPK(Thr172) was depressed in salt-loading adipocytes. The expression of pro-inflammatory adipocytokines, TNFα, MCP-1, COX-2, IL-17A, IL-6, leptin, and leptin to adiponectin ratio (LAR) were dose-dependently increased by salt treatment. Using the inhibitors of MAPK/ERK, U0126, we found that the crosstalk among the signaling pathways of MAPK/ERK, Akt-mTOR, and the inflammatory adipogenesis can be the possible mechanism of salt-linked obesity. The possibilities of whether the defense mechanisms against high dose of intracellular salts provoke signaling for adipocytes differentiation or interact with surrounding tissues through other pathways will be explored in future research

Targeted Next-Generation Sequencing at Copy-Number Breakpoints for Personalized Analysis of Rearranged Ends in Solid Tumors

<div><p>Background</p><p>The concept of the utilization of rearranged ends for development of personalized biomarkers has attracted much attention owing to its clinical applicability. Although targeted next-generation sequencing (NGS) for recurrent rearrangements has been successful in hematologic malignancies, its application to solid tumors is problematic due to the paucity of recurrent translocations. However, copy-number breakpoints (CNBs), which are abundant in solid tumors, can be utilized for identification of rearranged ends.</p><p>Method</p><p>As a proof of concept, we performed targeted next-generation sequencing at copy-number breakpoints (TNGS-CNB) in nine colon cancer cases including seven primary cancers and two cell lines, COLO205 and SW620. For deduction of CNBs, we developed a novel competitive single-nucleotide polymorphism (cSNP) microarray method entailing CNB-region refinement by competitor DNA.</p><p>Result</p><p>Using TNGS-CNB, 19 specific rearrangements out of 91 CNBs (20.9%) were identified, and two polymerase chain reaction (PCR)-amplifiable rearrangements were obtained in six cases (66.7%). And significantly, TNGS-CNB, with its high positive identification rate (82.6%) of PCR-amplifiable rearrangements at candidate sites (19/23), just from filtering of aligned sequences, requires little effort for validation.</p><p>Conclusion</p><p>Our results indicate that TNGS-CNB, with its utility for identification of rearrangements in solid tumors, can be successfully applied in the clinical laboratory for cancer-relapse and therapy-response monitoring.</p></div

Copy-number status information by cSNP microarray.

<p>The allele B is the B allele, and the values for various parameters are the expected values.</p><p>N-H mixture or N-H mix, the mixture of normal and H-mole DNAs; C-H mixture or C-H mix, the mixture of cancer and H-mole DNAs; N-H mix ratio, the ratio of normal and H-mole DNA amounts in the N-H mixture; C-H mix ratio, the ratio of cancer and H-mole DNA amounts in the C-H mixture; Copy Number, the copy number in cancer; Cancer Genotype, the genotype in cancer-sample DNA; Normal genotype, the genotype in normal-sample DNA; N-H mix genotype, the genotype in the N-H mixture; C-H mix genotype, the genotype in the C-H mixture; BAF_NH, the ratio of B allele and total allele amounts (or B allele frequency) in the N-H mixture; AR_NH, the ratio of B allele and A allele amounts (or allelic ratio) in the N-H mixture; BAF_CH, the ratio of B allele and total allele amounts (or B allele frequency) in the C-H mixture; AR_CH, the ratio of B allele and A allele amounts (or allelic ratio) in the C-H mixture; ARR, the ratio of AR_CH and AR_NH, or the AR ratio; nARR, the ARR values that are divided by the median ARR value, or normalized ARR.</p><p>The median ARR value (underlined) was used for normalization.</p

Copy-number changes at CNBs according to nARR values or LRRs.

<p>The copy-number statuses at nine CNBs in four primary colon cancer samples are shown. The CNBs were apparent with the nARR values, but the LRRs showed no clear breakpoints at most sites. The arrows indicate the CNBs employed for capture probes; at all of the CNBs marked by an arrow, the PCR-amplifiable tumor-specific rearrangements were successfully identified by TNGS-CNB.</p

PCR-amplified rearrangements identified by TNGS-CNB.

<p>*Chromosome number.</p><p>**Two rearrangements were amplified also in normal samples, indicating that these are constitutive genomic rearrangements.</p><p>***Two candidate rearrangements were analyzed by next-generation sequencing in the same region as sample C6, but only one was amplified by PCR.</p><p>****Samples C7 and C8 are the COLO205 and SW620 cancer cell lines, respectively.</p><p>ID, identification number.</p

TNGS-CNB schematic.

<p>The copy-number statuses of seven primary colon cancer tissues and two colon cancer cell lines were analyzed by the cSNP microarray method, and 91 CNBs were deduced. After removing 3.8 Mb of repetitive sequences from the CNB regions, the area for targeted capture was 2.2 Mb. After paired-end NGS of the captured sequences, the reads were aligned to the NCBI human genome assembly (build 37, hg19), and 23 candidate rearranged sequences were deduced. After PCR confirmation of the candidate rearranged sequences, 19 PCR-amplifiable rearrangements were identified.</p

cSNP microarray for determination of copy-number breakpoints.

<p>A. Schematic procedure for cSNP microarray. For each sample, SNP microarray experiments were performed for 1) H-mole DNA, 2) normal-sample DNA, 3) the mixture of normal and H-mole DNAs (N-H mixture), and 4) the mixture of cancer and H-mole DNAs (C-H mixture). An SNP microarray experiment for cancer samples was performed for the purpose of comparison. Only alter homozygous alleles (box) were selected for the cSNP microarray analysis. B. Representative LRR, nAR_CH, and nARR results for colon cancer samples. The LRR was obtained by Genome Studio software; the nAR_CH was calculated by cSNP software for the cancer and H-mole DNAs; the nARR was calculated by the same cSNP software for DNAs from the cancer tissues, corresponding normal tissues, and H-mole.</p

Preventive effects of protopanaxadiol and protopanaxatriol ginsenosides on liver inflammation and apoptosis in hyperlipidemic apoE KO mice

Ginsenosides, bioactive compounds of Panax GinsengC.A. Meyer, are divided into protopanaxadiol (PD) and protopanaxtriol (PT). The aim of this study was to evaluate the protective effects of different PD and PT combination ratios on liver inflammation and apoptosis in hyperlipidemic apo E KO mice. R1 (PD/PT = 1, high Rg1 and Rb1) and R2 (PD/PT = 2, high Re and Rd) extracts were intraperitoneally injected by 100 mg/kg/day at the 8th week. R1 and R2 improved atherogenic indices by increasing HDL and lowering total cholesterol (TC) and triacylglyceride (TG) selectively. R1 decreased lipid peroxides (LPO) level in plasma and liver tissue of hyperlipidemic mice, and R2 lowered plasma malondialdehyde(MDA) level. R1 and R2 not only regulated the expression of cyclooxygenase (COX)-2, IκB-α, phopho-ERK 1/2, and phopho-SAPK/JNK levels but also were significantly effective in blocking apoptotic signals, such as caspase-8, -9, as well as the cleavage of PARP in liver. Different combinational treatment of PD and PT extracts might ameliorate the liver inflammation and apoptosis in hyperlipidemic apo E KO mice, which is atherosclerotic animal model