Supplemental Material - Filial Caregiving and Chinese Adults’ Depressive Symptoms: Do Early-Life Parent-Child Relationships Matter?

Supplemental Material for Filial Caregiving and Chinese Adults’ Depressive Symptoms: Do Early-Life Parent-Child Relationships Matter? by Yue Qin, Jooyoung Kong, and Sara Moorman in Journal of Applied Gerontology</p

Supplemental Material - Lifetime Abuse Victimization and Prospective Health Outcomes in Older Adults

Supplemental Material for Lifetime Abuse Victimization and Prospective Health Outcomes in Older Adults by Jooyoung Kong, Sara M. Moorman, and Yue Qin in Journal of Applied Gerontology</p

Can Switching from Coal to Shale Gas Bring Net Carbon Reductions to China?

To increase energy security and reduce emissions of air pollutants and CO₂ from coal use, China is attempting to duplicate the rapid development of shale gas that has taken place in the United States. This work builds a framework to estimate the lifecycle greenhouse gas (GHG) emissions from China’s shale gas system and compares them with GHG emissions from coal used in the power, residential, and industrial sectors. We find the mean lifecycle carbon footprint of shale gas is about 30–50% lower than that of coal in all sectors under both 20 year and 100 year global warming potentials (GWP₂₀ and GWP₁₀₀). However, primarily due to large uncertainties in methane leakage, the upper bound estimate of the lifecycle carbon footprint of shale gas in China could be approximately 15–60% higher than that of coal across sectors under GWP₂₀. To ensure net GHG emission reductions when switching from coal to shale gas, we estimate the breakeven methane leakage rates to be approximately 6.0%, 7.7%, and 4.2% in the power, residential, and industrial sectors, respectively, under GWP₂₀. We find shale gas in China has a good chance of delivering air quality and climate cobenefits, particularly when used in the residential sector, with proper methane leakage control

Cu-Doped ZnInGaSe Nanocrystals with Controlled Stoichiometry for Green Emission

As one of Cd-free semiconductor nanocrystals, Zn-based nanocrystals are widely studied due to their wide band gap to be a candidate for blue- and green-emitting materials. Herein, ZnInGaSe nanocrystals were synthesized with the controlled stoichiometry from the intermediate of ZnInGaSe magic size clusters. The approach via the magic size clusters greatly increases the amount of Ga in ZnInGaSe NCs. After the growth of the ZnS shell, ZnInGaSe/ZnS core/shell nanocrystals exhibited improved green emission. Furthermore, Cu as the impurity was introduced into ZnInGaSe NCs to reduce the vacancy defects to promote the photoluminescence. Finally, a photoluminescence quantum yield of 35.7% was obtained in Cu:ZnInGaSe/ZnS core/shell nanocrystals. The donor–acceptor recombination mechanism was proposed to explain the optical properties in ZnInGaSe NCs. Cu:ZnInGaSe nanocrystals were utilized in light-emitting diodes, showing the potential of ZnInGaSe nanocrystals for optoelectronic devices

A multicomponent microemulsion using rational combination strategy improves lung cancer treatment through synergistic effects and deep tumor penetration

<p>Previously, we have developed a multicomponent-based microemulsion composed of etoposide, coix seed oil, and ginsenoside Rh2 (ECG-MEs). In this study, our goal was to validate the feasibility of ECG-MEs in lung cancer treatment and explore the mechanism underling the enhanced antitumor efficacy. The optimal weight ratio of ginsenoside Rh2 (G-Rh2) in ECG-MEs was determined as 3% (wt%), that was capable of forming the microemulsion readily with small particle size and high drug encapsulation efficiency. In cellular studies, the intracellular fluorescence of human non-small cell lung cancer (A549) cells treated with fluorescein isothiocyanate-labeled ECG-MEs (FITC/ECG-MEs) was significantly higher than that of various controls, leading to the obviously synergistic anticancer activities in cytotoxicity and <i>in vitro</i> cell apoptosis induction. The anticancer efficacy <i>in vivo</i> results showed that ECG-MEs markedly inhibited the growth of A549 tumor xenografts, potently induced tumor cells apoptosis, and obviously prolonged the survival time of mice. Of note, the mechanisms of enhanced anticancer efficiency were connected with the small size-mediated deep tumor penetration and increase in serum concentration of T helper 1 (Th1) cytokines. In summary, ECG-MEs exerting the rational drug combination strategy offers a solid evidence for lung cancer treatment, and has a promising potential for clinical application.</p

RNF26 Temporally Regulates Virus-Triggered Type I Interferon Induction by Two Distinct Mechanisms

<div><p>Viral infection triggers induction of type I interferons (IFNs), which are critical mediators of innate antiviral immune response. Mediator of IRF3 activation (MITA, also called STING) is an adapter essential for virus-triggered IFN induction pathways. How post-translational modifications regulate the activity of MITA is not fully elucidated. In expression screens, we identified RING finger protein 26 (RNF26), an E3 ubiquitin ligase, could mediate polyubiquitination of MITA. Interestingly, RNF26 promoted K11-linked polyubiquitination of MITA at lysine 150, a residue also targeted by RNF5 for K48-linked polyubiquitination. Further experiments indicated that RNF26 protected MITA from RNF5-mediated K48-linked polyubiquitination and degradation that was required for quick and efficient type I IFN and proinflammatory cytokine induction after viral infection. On the other hand, RNF26 was required to limit excessive type I IFN response but not proinflammatory cytokine induction by promoting autophagic degradation of IRF3. Consistently, knockdown of RNF26 inhibited the expression of <i>IFNB1</i> gene in various cells at the early phase and promoted it at the late phase of viral infection, respectively. Furthermore, knockdown of RNF26 inhibited viral replication, indicating that RNF26 antagonizes cellular antiviral response. Our findings thus suggest that RNF26 temporally regulates innate antiviral response by two distinct mechanisms.</p></div

RNF26 protects MITA from K48-linked polyubiquitination and degradation.

<p>(A) Effects of RNF26 knockdown on virus-induced polyubiquitination of endogenous MITA. The THP-1-RNF26-RNAi or control cells (2×10⁷) were infected with SeV or HSV-1 for the indicated time points or left uninfected. Cell lysates were subjected to IP under denatured conditions with anti-MITA and the immunoprecipitates were analyzed by immunoblots with anti-Ub(K48) (upper panel), anti-Ub(K63) (middle panel) or anti-MITA (lower panel). The whole cell lysates were analyzed by immunoblots with antibodies against the indicated proteins. (B) Effects of RNF5 knockdown on virus-induced K11-linked polyubiquitination of endogenous MITA. The 293-HA-Ub-K11O cells (2×10⁷) were transfected with the indicated RNAi plasmid (10 µg each). Twelve hours after transfection, the cells were selected with puromycin (1 µg/mL) for twenty-four hours and infected with SeV for the indicated time points or left uninfected. Cell lysates were subjected to IP under denatured conditions with anti-MITA and the immunoprecipitates were analyzed by immunoblots with anti-HA (upper panel) or anti-MITA (lower panel). The whole cell lysates were analyzed by immunoblots with antibodies against the indicated antibodies. (C) RNF26 and RNF5 competed with each other on MITA polyubiquitination. The 293 cells (5×10⁶) were transfected with MITA (5 µg), Flag-Ub-K48O and HA-Ub-K11O (1 µg each) together with indicated amount of RNF26 and RNF5. Twenty-four hours after transfection, cell lysates were subjected to IP under denatured conditions with anti-MITA and the immunoprecipitates were analyzed by immunoblots with anti-Flag (upper panel), anti-HA (middle panel) or anti-MITA (lower panel). The whole cell lysates were analyzed by immunoblots with antibodies against the indicated proteins. (D) Effects of RNF26 knockdown on virus-triggered MITA degradation. The THP-1-RNF26-RNAi or control cells (1×10⁶) were infected with SeV or HSV-1 for the indicated time points or left uninfected. Cells were lysed and whole cell lysates were analyzed by immunoblots with antibodies against the indicated proteins (upper immunoblots). The relative protein levels of MITA in reference to β-actin were analyzed by Quantity One program and data shown are mean ± S.D. of three independent experiments (lower histographs).</p

RNF26 catalyzes K11-linked polyubiquitination of MITA.

<p>(A and B) RNF26 targeted MITA for K11-linked polyubiquitination. The 293 cells (5×10⁶) were transfected with Flag-MITA (5 µg) and RNF26 (1 µg) together with HA-Ub or its mutants (1 µg each). Twenty-four hours after transfection, cells were subjected for IP under denatured conditions with limited amount of anti-Flag (0.5 µg) so that equal amount of Flag-MITA is pulled down. The immunoprecipitates were analyzed by immunoblots with anti-HA (upper panel) or anti-Flag (lower panel). The whole cell lysates were analyzed by immunoblots with anti-Flag or anti-RNF26 as indicated. Ub-AKR, all lysine residues of ubiquitin were mutated to arginine. (C) Effects of RNF26-RNAi plasmids on the expression of RNF26. In the upper panels, the 293 cells (1×10⁶) were transfected with the expression plasmids for RNF26-Flag (0.5 µg) and HA-β-actin (0.1 µg) together with the indicated RNAi plasmids (1 µg each). Twenty-four hours after transfection, whole cell lysates were analyzed by immunoblots with anti-Flag or anti-HA. In the lower panels, 293 cells were transduced with a control or RNF26-RNAi by retrovirus mediated gene transfer. Cells (1×10⁶) were lysed and whole cell lysates were analyzed by immunoblots with anti-RNF26 or anti-β-actin. (D) Immunoblot analysis of Flag-tagged ubiquitin expression in THP-1 cells stably transfected with Flag-Ub-K11O plasmid. Whole cell lysates of THP-1-Flag-Ub-K11O-RNF26-RNAi and control cells (1×10⁶) were analyzed by immunoblots with antibodies against the indicated proteins. RNF26-RNAi #1 was used here and in the following experiments if not noted. (E and F) Effects of RNF26 knockdown on virus-induced K11-linked polyubiquitination of endogenous MITA. In (E), THP-1-Flag-Ub-K11O-RNF26-RNAi or control cells (2×10⁷) were infected with SeV or HSV-1 for the indicated time points or left uninfected followed by IP under denatured conditions with anti-MITA. The immunoprecipitates were analyzed by immunoblots with anti-Flag (upper panels) or anti-MITA (lower panels). The whole cell lysates were analyzed by immunoblots with antibodies against the indicated cellular or viral proteins. In (F), 293-HA-Ub-K11O cells (2×10⁷) were transfected with a control or RNF26-RNAi plasmid (10 µg each). Twelve hours after transfection, puromycin (1 µg/mL) was added into the culture medium. The cells were selected for twenty-four hours and infected with SeV or left uninfected for the indicated time points followed by IP under denatured conditions and immunoblot analysis as in (E). All experiments were repeated for at least three times with similar results.</p

RNF26 promotes polyubiquitination of MITA at K150.

<p>(A) RNF26 mediated polyubiquitination of MITA at K150. The 293 cells (5×10⁶) were transfected with HA-Ub (1 µg) and RNF26 (1 µg) together with Flag-MITA or the indicated mutants (5 µg each). Twenty-four hours after transfection, cells were subjected IP under denatured conditions with anti-Flag and immunoprecipitates were analyzed by immunoblots with anti-HA (upper panel) or anti-Flag (lower panel). The whole cell lysates were analyzed by immunoblots with anti-Flag or anti-RNF26 as indicated. (B) RNF26 targeted MITA for polyubiquitination at K150 <i>in vitro</i>. RNF26, MITA and its mutants were obtained by <i>in vitro</i> transcription and translation. Biotin-Ub, E1, UbcH5 and RNF26 were incubated with MITA or its mutants. The ubiquitination of MITA was examined by immunoblot analysis with HRP-streptavidin (top panel). The inputs of RNF26 and MITA were analyzed by immunoblots with anti-MITA and anti-RNF26 (bottom panels). All experiments were repeated for at least three times with similar results.</p

RNF26 modulates virus-triggered induction of type I IFNs.

<p>(A) Effects of RNF26 knockdown on SeV-triggered activation of the IFN-β promoter. The 293 cells (2×10⁵) were transfected with the IFN-β promoter reporter (0.1 µg) and the indicated RNAi plasmid (0.5 µg each). Thirty hours after transfection, cells were infected with SeV for 12 hours or left uninfected before reporter assays were performed. (B and C) Effects of RNF26 knockdown on virus-triggered induction of IFN-β in THP-1 cells. The THP-1-RNF26-RNAi or control cells (1×10⁶) were infected with SeV or HSV-1 for the indicated time points or left uninfected followed by quantitative real-time PCR (B) or ELISA analysis (C). ND, not detected. (D) Effects of RNF26 knockdown on virus-triggered induction of <i>IFNB1</i> gene in THP-1 cells. The THP-1-RNF26-RNAi or control cells (1×10⁶) were infected with VSV, EMCV or ECTV for the indicated time points or left uninfected before quantitative real-time PCR analysis was performed as in (B). (E and F) Effects of RNF26 knockdown on virus-triggered induction of TNFα in THP-1 cells. The THP-1-RNF26-RNAi or control cells (1×10⁶) were infected with SeV or HSV-1 for the indicated time points or left uninfected followed by quantitative real-time PCR (E) and ELISA analysis (F). (G) Effects of RNF26 knockdown on virus-triggered phosphorylation of TBK1, IRF3 and IκBα. The THP-1-RNF26-RNAi or control cells (1×10⁶) were infected with SeV or HSV-1 for the indicated time points or left uninfected, whole cell lysates were analyzed by immunoblots with anti-p-TBK1, anti-TBK1, anti-p-IRF3, anti-IRF3, anti-p-IκBα, anti-IκBα, anti-RNF26 or anti-β-actin as indicated. All experiments were repeated for at least three times with similar results. The bar graphs show mean ± S.D. (<i>n</i> = 3) of a representative experiment performed in triplicate.</p