Minimally modified LDL upregulates endothelin type B receptors in rat coronary artery via ERK1/2 MAPK and NF-kappa B pathways

Minimally modified low density lipoprotein (mmLDL) is a well-known risk factor for coronary artery disease. Upregulation of vascular endothelin type B (ETB) receptors on the vascular smooth muscle cells is predicted to be the molecular mechanism that leads to cardiovascular pathogenesis. The objective of the present study was to examine the hypothesis that mmLDL upregulates ETB receptors in rat coronary artery. The contractile responses to sarafotoxin 6c (ETB receptor agonist) were studied using a sensitive myograph. ETB receptor mRNA and protein expression was determined using real-time PCR and Western blot analysis. The results showed that organ culture increased the contractile responses induced by sarafotoxin 6c and the levels of ETB receptor mRNA and protein. This increase was further enhanced by the addition of mmLDL (10 mu g/mL). Specific ERK1/2 inhibitors (SB386023 and U0126) and an NF-kappa B inhibitor (wedelolactone) attenuated the mmLDL-increased ETB receptor-mediated contraction and ETB receptor mRNA and protein levels. Wedelolactone significantly attenuated the mmLDL-decreased I kappa B-alpha protein expression. Consistent with this result, I kappa B-alpha protein expression was significantly decreased by culture with mmLDL compared to the level of expression in the organ culture group. However, the JNK inhibitor, SP600125 or p38 pathway inhibitor, SB203580 did not inhibit mmLDL-enhanced effects. The PKC inhibitor, staurosporine attenuated only culture-alone-increased effects. In conclusion, mmLDL upregulates the ETB receptors in rat coronary arterial smooth muscle cells, mainly via activation of the ERK1/2 MAPK and the downstream transcriptional factor NF-kappa B. (C) 2011 Elsevier B.V. All rights reserved

Secondhand smoke exposure induces Raf/ERK/MAPK-mediated upregulation of cerebrovascular endothelin ETA receptors

<p>Abstract</p> <p>Background</p> <p>Cigarette smoking enhances the risk of stroke. However, the underlying molecular mechanisms are largely unknown. The present study established an <it>in vivo </it>rat secondhand cigarette smoking (SHS) model and examined the hypothesis that SHS upregulates endothelin receptors with increased cerebrovascular contraction <it>via </it>the Raf/extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinases (MAPK) pathway.</p> <p>Results</p> <p>Rats were exposed to SHS for up to 8 weeks. The cerebral artery vasoconstriction was recorded by a sensitive myograph. The mRNA and protein expressions for endothelin receptors in cerebral arteries were studied by real-time PCR and Western blot. Compared to fresh air exposed rats, cerebral arteries from SHS rats exhibited stronger contractile responses (<it>P </it>< 0.05) mediated by endothelin type A (ET_A) receptors. The expressions of mRNA and protein for ET_Areceptors in the cerebral arteries from SHS rats were higher (<it>P </it>< 0.05) than that in control. SHS did not affect endothelin type B (ET_B) receptor-mediated contractions, mRNA or protein levels. The results suggest that SHS upregulates ET_A, but not ET_Breceptors <it>in vivo</it>. After SHS exposure, the mRNA levels of Raf-1 and ERK1/2, the protein expression of phosphorylated (p)-Raf-1 and p-ERK1/2 were increased (<it>P </it>< 0.05). Raf-1 inhibitor, GW5074 suppressed the enhanced ET_Areceptor-mediated contraction, mRNA and protein levels induced by SHS. In addition, GW5074 inhibited the SHS-caused increased mRNA and phosphorylated protein levels of Raf-1 and ERK1/2, suggesting that SHS induces activation of the Raf/ERK/MAPK pathway.</p> <p>Conclusions</p> <p>SHS upregulates cerebrovascular ET_Areceptors <it>via </it>the Raf/ERK/MAPK pathway, which provides novel understanding of mechanisms involved in SHS-associated stroke.</p

Abl Family Tyrosine Kinases Are Essential for Basement Membrane Integrity and Cortical Lamination in the Cerebellum

The Abl family nonreceptor tyrosine kinases, consisting of closely related Abl and Arg (Abl-related gene), play essential roles in mouse neurulation, but their functions in the subsequent development of CNS are poorly understood. Here, we show that conditional deletion of Abl in precursors of neurons and glia on an Arg knock-out background leads to striking cerebellar malformations, including defects in anterior cerebellar morphogenesis, granule cell ectopia, and hypoplasia. Time course analyses reveal that the abnormal anterior cerebellar foliation results from local disruptions of the basement membrane (BM) located between radial glial endfeet and the meninges during embryonic cerebellar development. Granule cell ectopia and hypoplasia are also associated with the breaches in the BM and abnormal Bergmann glial networks during postnatal cerebellar development. In vitro culture experiments indicate that Abl/Arg-deficient granule cells can interact with glial processes and proliferate normally in response to sonic hedgehog compared to cells isolated from control mice. Consistent with these findings, selective ablation of Abl family kinases in cerebellar granule cells alone does not cause any abnormality, suggesting that deletion of Abl/Arg from glia is likely required for the mutant phenotype. Together, these results provide compelling evidence that Abl and Arg play key redundant roles in BM maintenance and cortical lamination in the cerebellum

The CUL4-DDB1 ubiquitin ligase complex controls adult and embryonic stem cell differentiation and homeostasis

Little is known on post-transcriptional regulation of adult and embryonic stem cell maintenance and differentiation. Here we characterize the role of Ddb1, a component of the CUL4-DDB1 ubiquitin ligase complex. Ddb1 is highly expressed in multipotent hematopoietic progenitors and its deletion leads to abrogation of both adult and fetal hematopoiesis, targeting specifically transiently amplifying progenitor subsets. However, Ddb1 deletion in non-dividing lymphocytes has no discernible phenotypes. Ddb1 silencing activates Trp53 pathway and leads to significant effects on cell cycle progression and rapid apoptosis. The abrogation of hematopoietic progenitor cells can be partially rescued by simultaneous deletion of Trp53. Conversely, depletion of DDB1 in embryonic stem cell (ESC) leads to differentiation albeit negative effects on cell cycle and apoptosis. Mass spectrometry reveals differing protein interactions between DDB1 and distinct DCAFs, the substrate recognizing components of the E3 complex, between cell types. Our studies identify CUL4-DDB1 complex as a novel post-translational regulator of stem and progenitor maintenance and differentiation

Gold-Catalyzed Tandem Cycloisomerizationand Dimerization of Chiral Homopropargyl Sulfonamides

We are grateful for the financial support from the National Natural Science Foundation of China (No. 21102119 and 21272191), the Natural Science Foundation of Fujian Province of China (No. 2012J01051), NFFTBS (No. J1210014), and PCSIRT. We also thank Professor Dr. Liming Zhang (University of California Santa Barbara) for the helpful discussions

Lipid-soluble smoke particles damage endothelial cells and reduce endothelium-dependent dilatation in rat and man

BACKGROUND: Cigarette smoking is a strong risk factor for vascular disease and known to cause dysfunction of the endothelium. However, the molecular mechanisms involved are still not fully understood. METHODS: In order to reveal the direct effects of lipid-soluble smoke particles on the endothelium, ring segments isolated from rat mesenteric arteries and human middle cerebral arteries (MCA) obtained at autopsy were incubated for 6 to 48 hrs in the presence of dimethylsulphoxide (DMSO)-soluble particles from cigarette smoke (DSP), i.e. lipid-soluble smoke particles. The endothelial microstructure was examined by transmission electron microscopy. The endothelial function was evaluated by acetylcholine (ACh)-induced endothelium-dependent vasodilatation, using a sensitive myograph. RESULTS: After DSP treatment, the arterial endothelium was swollen and loosing its attachment. In functional tests, the total ACh-induced dilatation, the nitric oxide (NO)-mediated and the endothelium-derived hyperpolarization factor (EDHF)-mediated dilatations were significantly decreased by DSP in a time- and concentration-dependent manner (p < 0.05). Nicotine, an important compound in cigarette smoke had, in an equivalent concentration as in DSP, no such effects (p > 0.05). Similar results were obtained in the human MCA. CONCLUSION: Thus, we demonstrate that the lipid-soluble smoke particles, but not nicotine, caused damage to arterial endothelium and reduced the endothelium-dependent dilatation in man and rat

Phase II of the LAMOST-Kepler/K2 survey. I. Time series of medium-resolution spectroscopic observations

Phase \RNum{2} of the LAMOST-{\sl Kepler/K}2 survey (LK-MRS), initiated in 2018, aims at collecting medium-resolution spectra ($R\sim7,500$; hereafter MRS) for more than $50,000$ stars with multiple visits ($\sim60$ epochs) over a period of 5 years (2018 September to 2023 June). We selected 20 footprints distributed across the {\sl Kepler} field and six {\sl K}2 campaigns, with each plate containing a number of stars ranging from $\sim2,000$ to $\sim 3,000$. During the first year of observations, the LK-MRS has already collected $\sim280,000$ and $\sim369,000$ high-quality spectra in the blue and red wavelength range, respectively. The atmospheric parameters and radial velocities for $\sim259,000$ spectra of $21,053$ targets were successfully calculated by the LASP pipeline. The internal uncertainties for the effective temperature, surface gravity, metallicity, and radial velocity are found to be $100$\,K, $0.15$\,dex, $0.09$\,dex, and $1.00$\,km\,s$^{-1}$, respectively. We found $14,997$, $20,091$, and $1,514$ stars in common with the targets from the LAMOST low-resolution survey (LRS), GAIA and APOGEE, respectively, corresponding to a fraction of $\sim70\%$, $\sim95\%$ and $\sim7.2\%$. In general, the parameters derived from LK-MRS spectra are consistent with those obtained from the LRS and APOGEE spectra, but the scatter increases as the surface gravity decreases when comparing with the measurements from APOGEE. A large discrepancy is found with the GAIA values of the effective temperature. The comparisons of radial velocities of LK-MRS to GAIA and LK-MRS to APOGEE nearly follow an Gaussian distribution with a mean $\mu\sim1.10$ and $0.73$\,km\,s$^{-1}$, respectively.Comment: 24 pages, 15 figures, 4 tables, ApJS, accepte

Genetic Abolishment of Hepatocyte Proliferation Activates Hepatic Stem Cells

Quiescent hepatic stem cells (HSCs) can be activated when hepatocyte proliferation is compromised. Chemical injury rodent models have been widely used to study the localization, biomarkers, and signaling pathways in HSCs, but these models usually exhibit severe promiscuous toxicity and fail to distinguish damaged and non-damaged cells. Our goal is to establish new animal models to overcome these limitations, thereby providing new insights into HSC biology and application. We generated mutant mice with constitutive or inducible deletion of Damaged DNA Binding protein 1 (DDB1), an E3 ubiquitin ligase, in hepatocytes. We characterized the molecular mechanism underlying the compensatory activation and the properties of oval cells (OCs) by methods of mouse genetics, immuno-staining, cell transplantation and gene expression profiling. We show that deletion of DDB1 abolishes self-renewal capacity of mouse hepatocytes in vivo, leading to compensatory activation and proliferation of DDB1-expressing OCs. Partially restoring proliferation of DDB1-deficient hepatocytes by ablation of p21, a substrate of DDB1 E3 ligase, alleviates OC proliferation. Purified OCs express both hepatocyte and cholangiocyte markers, form colonies in vitro, and differentiate to hepatocytes after transplantation. Importantly, the DDB1 mutant mice exhibit very minor liver damage, compared to a chemical injury model. Microarray analysis reveals several previously unrecognized markers, including Reelin, enriched in oval cells. Here we report a genetic model in which irreversible inhibition of hepatocyte duplication results in HSC-driven liver regeneration. The DDB1 mutant mice can be broadly applied to studies of HSC differentiation, HSC niche and HSCs as origin of liver cancer