Abstract Cross-priming of CD8+ T cells and generation of effector immune responses is pivotal for tumor immunity, successful anticancer vaccination and therapy. We have discovered a novel molecular mechanism operating in antigen donor cells, which regulates cross-priming of CD8+ T cells during primary sterile necrosis and thereby controls adaptive immune responses. We show that cellular peptidases, dipeptidyl peptidase 3 (DPP-3) and thimet oligopeptidase 1 (TOP-1) present in non-immunogenic necrotic cells eliminate proteasomal degradation products and block antigen cross-presentation. While sterile necrotic tumor cells fail to induce CD8+ T cell responses, their non-immunogenicity can be reversed in vitro and in vivo by inactivation of peptidases, DPP-3 and TOP-1. Thus control of cross-priming and thereby immunogenicity of primary sterile necrosis relies on the proteasome dependent oligopeptide generation and functional status of peptidases in antigen donor cells. Citation Format: Jaba Gamrekelashvili, Tamar Kapanadze, Josef Wissing, Chi Ma, Lothar Jaensch, Firouzeh Korangy, Tim F. Greten. Cross-priming of CD8+ T cells is controlled by dipeptidyl peptidase 3 and thimet oligopeptidase 1 present in necrotic cells. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B30.
The combined effects of climate change and human activities pose threat to the sustainable development of ecosystems. Human appropriation of net primary production (HANPP) has been extensively used as an important indicator for evaluating the sustainable development of the ecosystem. However, few studies quantitatively assessed the driving factors of HANPP. Based on Moderate-Resolution Imaging Spectroradiometer (MODIS) data, methods of net primary production (NPP) model and regression analysis, the spatial and temporal distribution of HANPP and its driving factors in the Qilian Mountains from 2005 to 2015 were illustrated. The results showed that the HANPP in the Qilian Mountains decreased gradually from both east to west and from south to north, showing a slight upward overall. The regions affected by human activities and climate change accounted for 26.8% and 73.2% respectively. Moreover, there was a significant negative impact between grain yield and HANPP, and a significant positive impact of either the annual sunshine duration, or livestock amount to HANPP.
Objective
To investigate the effect and mechanism of ubiquitin-specific processing peptidase 10 (USP10) on gemcitabine resistance of pancreatic cancer.
Methods
(1) The 50% inhibiting concentration (IC50) of USP10 in the PANC-1, BxPC-3 and SW1990 cell lines of pancreatic cancer was detected by CCK8 assay. (2) The expression of USP10 in the BxPC-3, PANC-1, SW1990 and SW1990-GEM cell lines of pancreatic cancer was detected by Western blot. (3) Small interfering RNA (siRNA) of SW1900 cell lines was conducted, siRNA and USP10siRNA were transected and then were respectively allocated into the control group and USP10 group. (4) IC50 of gemcitabine in the cancer cell lines between the 2 groups was detected by CCK8 assay. (5) Colony-forming unit assay: number of cloned cells was counted and colony-forming efficiency (CFE) was calculated. (6) Cells in the 2 groups were interfered with 0.3 μg/mL gemcitabine for 48 hours, and cell apoptosis was detected by flow cytometry and percentage of apoptosis was calculated. (7) Cells in the 2 groups were interfered with 0.3 μg/mL gemcitabine for 48 hours, and cell cycle was detected by flow cytometry. (8) The relative expressions of proteins in the 2 groups were detected by Western blot: ① the relative expressions of USP10 and P21 of cell lines interfered with 0, 0.30, 0.60, 1.20 μg/mL gemcitabine for 72 hours in the 2 groups were detected. ② The relative expressions of USP10 and P21 of cell lines interfered with 0.30 μg/mL gemcitabine at 0, 24, 48, 72 hours in the 2 groups were detected. ③ The relative expression of P53 in SW1990 and SW1990-GEM cell lines was detected. ④ The relative expression of P53 in SW1990 cell line interfered with 0, 0.30, 0.60, 1.20 μg/mL gemcitabine for 72 hours was detected. ⑤ The relative expression of P53 in SW1990 cell line interfered with 0.30 μg/mL gemcitabine at 0, 24, 48, 72 hours was detected. (9) The interaction between the proteins was detected by co-immunoprecipitation. Measurement data was presented as ±s. The comparisons among groups were evaluated with the one-way ANOVA, and parirwise comparison was analyzed by the LSD test. Comparison between groups was analyzed by t test.
Results
(1) The IC50 of gemcitabine and relative expression of USP10 in the cancer cell lines: ① the IC50 of BxPC-3, PANC-1 and SW1990 cell lines interfered with gemcitabine for 72 hours which was detected using CCK8 assay was (0.070±0.040)μg, (0.120±0.010)μg and (0.350±0.050)μg, with a statistically significant difference among them (F=10.765, P 0.05), and there was a statistically significant difference in the expression of P21 between the 2 groups (t=3.765, P 0.05). ③ The relative expressions of P53 in SW1990 and SW1990-GEM cell lines were 2.650±0.050 and 1.450±0.060, respectively, showing a statistically significant difference (t=19.075, P<0.05). The relative expressions of P53 in the control and USP10 groups were 3.250±0.050 and 1.550±0.050, respectively, showing a statistically significant difference (t=9.240, P<0.05). ④ The relative expressions of P53 in SW1990 cell line interfered with 0, 0.30, 0.60, 1.20 μg/mL gemcitabine for 72 hours were 0.590±0.050, 1.015±0.050, 2.050±0.050 and 2.850±0.050, with a significant concentration-dependence (F=34.088, P<0.05). ⑤ The relative expressions of P53 in SW1990 cell line interfered with 0.30 μg/mL gemcitabine at 0, 24, 48, 72 hours were 0.890±0.050, 1.225±0.030, 2.180±0.150 and 3.030±0.150, with a significant time dependence (F=29.650, P<0.05). (7) The results of co-immunoprecipitation: there was expression of USP10 protein in P53 protein complexes and expression of P53 protein in USP10 protein complexes.
Conclusion
USP10 promotes primary and acquired resistance of gemcitabine in pancreatic cancer via activation of P53/P21 pathway.
Key words:
Pancreatic neoplasms; Gemcitabine; Resistance
Antibiotic tolerance poses a threat to current antimicrobial armamentarium. Bacteria at a tolerant state survive in the presence of antibiotic treatment and account for persistence, relapse and recalcitrance of infections. Antibiotic treatment failure may occur due to antibiotic tolerance. Persistent infections are difficult to treat and are often associated with poor prognosis, imposing an enormous burden on the healthcare system. Effective strategies targeting antibiotic-tolerant bacteria are therefore highly warranted. In this study, small molecule compound SA-558 was identified to be effective against Staphylococcus aureus that are tolerant to being killed by conventional antibiotics. SA-558 mediated electroneutral transport across the membrane and led to increased ATP and ROS generation, resulting in a reduction of the population of antibiotic-tolerant bacteria. In a murine chronic infection model, of which vancomycin treatment failed, we demonstrated that SA-558 alone and in combination with vancomycin caused significant reduction of MRSA abundance. Our results indicate that SA-558 monotherapy or combinatorial therapy with vancomycin is an option for managing persistent S. aureus bacteremia infection and corroborate that bacterial metabolism is an important target for counteracting antibiotic tolerance.
UL16-binding proteins [ULBPs, also termed as retinoic acid early transcripts (RAET1) molecules] are frequently expressed by malignant transformed cells and stimulate anti-tumor immune responses mediated by NKG2D-positive NK cells, CD8+ αβ T cells and γδ T cells in vitro and in vivo. In this study, we identified four novel functional splice variants of ULBPs including ULBP4-I, ULBP4-II, ULBP4-III and RAET1G3 in HepG2 liver carcinoma cells, WISH human amnion cells, Hep-2 larynx carcinoma cells and K562 leukemia cells, respectively, by reverse transcription–PCR and T vector cloning strategy. Analysis of alignments of amino acid sequences of the splice variants illustrated that there were important modifications between splice variants and their individual parental ULBP. All ULBP4 splice variants (ULBP4-I, ULBP4-II and ULBP4-III) were type 1 membrane-spanning molecules and had the ability to bind with human NKG2D receptor in vitro. Ectopic expressions of ULBP4 and ULBP4 splice variants resulted in the enhanced cytotoxic sensitivity of target cells against NK cells, which could be blocked by anti-NKG2D mAb. Moreover, co-culture-free soluble forms of ULBP4 splice variants (their α1 + α2 ectodomains) and RAET1G3 (soluble splice variant of RAET1G2) with NK cells down-regulated the cell surface expression of NKG2D. Finally, immobilized in a plate-bound form of RAET1G3 stimulated NK cells to secrete IFN-γ. Taken together, all the identified functional splice variants will help to advance our knowledge regarding the overall functions of ULBP gene family.
Objective In China, despite a high coverage rate, health insurance is not used for all illness episodes. Our goal is to identify subjects' characteristics associated with insurance utilization and the association between utilization and medical expenditure. Methods A survey was conducted in January and February of 2012. 2093 middle-aged and elderly subjects (45 years old and above) were surveyed. Results Heath insurance was not utilized for 12.6% (inpatient), 53.3% (outpatient), and 72.6% (self-treatment) of disease episodes. Subjects' characteristics were associated with insurance utilization. Inpatient and outpatient treatments were expensive. In the multivariate analysis of outpatient treatment expenditure, insurance utilization was significantly associated with higher treatment cost, lost income, and gross total cost. Conclusion Utilization of health insurance may need to be improved. Insurance utilization can reduce out-of-pocket medical expenditure. However, the amount paid by the insured is still high. Policy intervention is needed to further improve the effectiveness of health insurance.
Taking landslide with accumulation layer-bedrock contact surface which located in Langao county in Shaanxi province as the research object, the basic characters of landslides are summarized by field investigating, engineering geological analyzing and mathematical statistics method, the main characters can be concluded as follows:(1)those landslides mainly distribute in 600 m~ 1200 m elevation, slope forms are mainly convex with 180~ 270 ° dips and 20~40 ° gradient, meanwhile, those landslides distribute along the tectonic zone; (2) this kind of landslide is always shallow landslide, their property are mainly composed by crushed silty clay or gravel soil, The sliding form is mainly arc and line, which is easy to form "debris flow" during the sliding process;(3) the main failure mode are creep-rupture and slip-push.This research has guiding significance to reveal the formation mechanism and disaster prevention of landslide with accumulation layer-bedrock contact surface.
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