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Novel therapeutic approaches combining immune checkpoint inhibitors are needed to improve clinical outcomes for patients with cancer. Lymphocyte-activation gene 3 (LAG-3) is an immune checkpoint molecule that inhibits T-cell activity and antitumor immune responses, acting through an independent mechanism from that of programmed death-1 (PD-1) and cytotoxic T lymphocyte associated antigen-4 (CTLA-4). Here, we describe the development and preclinical characterization of relatlimab, a human antibody that binds to human LAG-3 with high affinity and specificity to block the interaction of LAG-3 with the ligands MHC II and fibrinogen like protein-1, and to reverse LAG-3–mediated inhibition of T-cell function in vitro. Consistent with previous reports, in mouse models, the combined blockade of LAG-3 and PD-1 with surrogate antibodies resulted in enhanced anti-tumor activity greater than the individual blockade of either receptor. In toxicity studies in cynomolgus monkeys, relatlimab was generally well tolerated when combined with nivolumab. These results are consistent with findings from the RELATIVITY-047 phase 2/3 trial showing that relatlimab combined with nivolumab is a well-tolerated regimen that demonstrated superior progression-free survival compared with nivolumab monotherapy in patients with unresectable or metastatic melanoma.SynopsisPreclinical studies demonstrate that relatlimab specifically blocks the interaction between LAG-3 and its ligands, and provide a biological rationale for combining relatlimab with the anti−PD-1 antibody nivolumab as an effective cancer immunotherapeutic strategy.

Novel therapeutic approaches combining immune-checkpoint inhibitors are needed to improve clinical outcomes for patients with cancer. Lymphocyte-activation gene 3 (LAG-3) is an immune-checkpoint molecule that inhibits T-cell activity and antitumor immune responses, acting through an independent mechanism from that of programmed death-1 (PD-1) and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4). Here, we describe the development and preclinical characterization of relatlimab, a human antibody that binds to human LAG-3 with high affinity and specificity to block the interaction of LAG-3 with the ligands MHC II and fibrinogen-like protein-1, and to reverse LAG-3–mediated inhibition of T-cell function in vitro. Consistent with previous reports, in mouse models, the combined blockade of LAG-3 and PD-1 with surrogate antibodies resulted in enhanced antitumor activity greater than the individual blockade of either receptor. In toxicity studies in cynomolgus monkeys, relatlimab was generally well tolerated when combined with nivolumab. These results are consistent with findings from the RELATIVITY-047 phase II/III trial showing that relatlimab combined with nivolumab is a well-tolerated regimen that demonstrates superior progression-free survival compared with nivolumab monotherapy in patients with unresectable or metastatic melanoma.

Supplementary Data from Preclinical Characterization of Relatlimab, a Human LAG-3–Blocking Antibody, Alone or in Combination with Nivolumab

PDF file - 25K, Figure S5. Effect of nivolumab on CD8+ T-cell subsets and DCs in PBMCs from nivolumab-treated cynomolgus monkeys.

PDF file - 376K, Figure S2. Epitope of nivolumab on human PD-1.

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PDF file - 22K, Figure S3. Nivolumab inhibits PD-1 interaction with PD-L1 and PD-L2.

Small cell lung carcinoma (SCLC) is highly mutated, yet durable response to immune checkpoint blockade (ICB) is rare. SCLC also exhibits cellular plasticity, which could influence its immunobiology. Here we discover that a distinct subset of SCLC uniquely upregulates MHC I, enriching for durable ICB benefit. In vitro modeling confirms epigenetic recovery of MHC I in SCLC following loss of neuroendocrine differentiation, which tracks with derepression of STING. Transient EZH2 inhibition expands these nonneuroendocrine cells, which display intrinsic innate immune signaling and basally restored antigen presentation. Consistent with these findings, murine nonneuroendocrine SCLC tumors are rejected in a syngeneic model, with clonal expansion of immunodominant effector CD8 T cells. Therapeutically, EZH2 inhibition followed by STING agonism enhances T-cell recognition and rejection of SCLC in mice. Together, these data identify MHC I as a novel biomarker of SCLC immune responsiveness and suggest novel immunotherapeutic approaches to co-opt SCLC's intrinsic immunogenicity.Significance:SCLC is poorly immunogenic, displaying modest ICB responsiveness with rare durable activity. In profiling its plasticity, we uncover intrinsically immunogenic MHC Ihi subpopulations of nonneuroendocrine SCLC associated with durable ICB benefit. We also find that combined EZH2 inhibition and STING agonism uncovers this cell state, priming cells for immune rejection.This article is highlighted in the In This Issue feature, p. 1861

PDF file - 239K, Figure S4. A, Expression of PD-1 in CD4 T cells during mixed lymphocyte reaction (MLR). B, Expression of PD-1 and PD-L1 during stimulation of peripheral blood mononuclear cells (PBMC) by superantigen SEB.

Supplementary Table S1

Supplementary Figures 1-12

PDF file - 72K, Figure S1. Binding of nivolumab to human T cells.

Purpose: The ganglioside fucosyl-GM1 (FucGM1) is a tumor-associated antigen expressed in a large percentage of human small cell lung cancer (SCLC) tumors, but absent in most normal adult tissues, making it a promising target in immuno-oncology. This study was undertaken to evaluate the preclinical efficacy of BMS-986012, a novel, nonfucosylated, fully human IgG1 antibody that binds specifically to FucGM1.Experimental Design: The antitumor activity of BMS-986012 was evaluated in in vitro assays using SCLC cells and in mouse xenograft and syngeneic tumor models, with and without chemotherapeutic agents and checkpoint inhibitors.Results: BMS-986012 showed a high binding affinity for FcγRIIIa (CD16), which resulted in enhanced antibody-dependent cellular cytotoxicity (ADCC) against FucGM1-expressing tumor cell lines. BMS-986012–mediated tumor cell killing was also observed in complement-dependent cytotoxicity (CDC) and antibody-dependent cellular phagocytosis (ADCP) assays. In several mouse SCLC models, BMS-986012 demonstrated efficacy and was well tolerated. In the DMS79 xenograft model, tumor regression was achieved with BMS-986012 doses of 0.3 mg/kg and greater; antitumor activity was enhanced when BMS-986012 was combined with standard-of-care cisplatin or etoposide. In a syngeneic model, tumors derived from a genetically engineered model of SCLC were treated with BMS-986012 or anti-FucGM1 with a mouse IgG2a Fc and their responses evaluated; when BMS-986012 was combined with anti–PD-1 or anti-CD137 antibody, therapeutic responses significantly improved.Conclusions: Single-agent BMS-986012 demonstrated robust antitumor activity, with the addition of chemotherapeutic or immunomodulatory agents further inhibiting SCLC growth in the same models. These preclinical data supported evaluation of BMS-986012 in a phase I clinical trial of patients with relapsed, refractory SCLC. Clin Cancer Res; 24(20); 5178–89. ©2018 AACR.

4 suppl figures on tumor models

4 suppl tables on FucGM1 expression in lung cancer, binding, and mouse PK

To investigate the effect of corosolic acid treatment on erectile function in metabolic syndrome induced rat model. Fifty male 3-week-old SD rats were fed a high fat and high sugar diet. Six months later, metabolic variables were determined. Metabolic syndrome induced erectile function (MED) rats were confirmed by an apomorphine test. Then MED rats were treated with corosolic acid daily by oral gavage for 4 weeks. To evaluate erectile function, intracavernosal pressure (ICP)/mean arterial blood pressure (MAP) ratio was measured. Thiobarbituric acid reactive substances assay and dihydroethidium staining were used to assess reactive oxygen species (ROS) level. Protein expressions of gp91phox and eNOS were examined by western blotting and immunohistochemistry. Fasting blood glucose, body weight, total cholesterol and insulin were markedly increased in metabolic syndrome rats compared with those of the control rats (p < 0.05). The ratios of max ICP/MAP and area under curve (AUC)/MAP was markedly reduced in MED rats compared with the control rats (p < 0.05). The concentration of cyclic guanosine mono-phosphate (cGMP) and the expression of eNOS were significantly decreased in MED rats compared with the control group (p < 0.05). Moreover, ROS level and the expression of gp91phox were significantly increased in MED rats. Treatment with corosolic acid reversed these changes (each p < 0.05). Corosolic acid reduces the level of ROS, ameliorating endothelial dysfunction and improvement of erectile function in MED rats.

Additional file 1: Table A1. Thirty-nine components of Solanum nigrum and their corresponding molecular weight (MW), predicted oral bioavailability (OB) and drug likeness (DL) scores. Table A2. Bladder cancer (BC) related targets in Solanum nigrum. By combining the related targets of active ingredients screened from S. nigrum and the disease-related targets, 100 overlapping targets (involving 23 duplicates) were selected as the key targets involved in the treatment of BC. Table A3. Gene ontology (GO) terms of therapeutic target genes and their corresponding counts, p-value, and FDR. Through the GO enrichment of the key targets, 86 remarkably enriched (p-value ≤0.01) GO terms were obtained, indicating that some targets were involved in tumorigenesis. Table A4. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of therapeutic target genes and their corresponding counts, p-value, and FDR. Through KEGG enrichment of the key targets, 89 remarkably enriched (p-value ≤0.01) pathways were obtained, indicating that numerous targets are involved in the processes of tumorigenesis and tumor progression.

Despite huge promises, bioanalysis of protein biomarkers in formalin-fixed paraffin-embedded (FFPE) tissues by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for clinical applications is still very challenging. Here, we describe a sensitive and robust LC-MS/MS assay to quantify clinical protein biomarkers in FFPE tumor sections using automated antipeptide antibody immunocapture followed by in-sample calibration curve (ISCC) strategy with multiple isotopologue reaction monitoring (MIRM) technique. ISCC approach with MIRM of stable isotopically labeled (SIL) peptides eliminated the need for authentic matrices for external calibration curves, overcame the matrix effects, and validated the quantification range in each individual sample. Specifically, after deparaffinization, rehydration, antigen retrieval, and homogenization, the protein analytes in FFPE tumor tissues were spiked with a known concentration of one SIL peptide for each analyte, followed by trypsin digestion and antipeptide immunocapture enrichment prior to MIRM-ISCC-based LC-MS/MS analysis. This approach has been successfully used for sensitive quantification of programmed cell death-1 (PD-1) and programmed cell death-ligand 1 (PD-L1) in 15 representative FFPE tumor samples from lung, colorectal, and head and neck cancer patients. Except for one sample, PD-L1 and PD-1 in all samples were quantifiable using this assay with concentrations of 27.85-798.43 (amol/μg protein) for PD-L1 and 16.96-129.89 (amol/μg protein) for PD-1. These results were generally in agreement with the immunohistochemistry (IHC) data but with some exceptions. This approach demonstrated the feasibility to quantify low abundant protein biomarkers in FFPE tissues with improved sensitivity, specificity, and robustness and showed great potential as an orthogonal analytical approach to IHC for clinical applications.

Purpose: The ganglioside fucosyl-GM1 (FucGM1) is a tumor-associated antigen expressed in a large percentage of human small cell lung cancer (SCLC) tumors, but absent in most normal adult tissues, making it a promising target in immuno-oncology. This study was undertaken to evaluate the preclinical efficacy of BMS-986012, a novel, nonfucosylated, fully human IgG1 antibody that binds specifically to FucGM1. Experimental Design: The antitumor activity of BMS-986012 was evaluated in in vitro assays using SCLC cells and in mouse xenograft and syngeneic tumor models, with and without chemotherapeutic agents and checkpoint inhibitors. Results: BMS-986012 showed a high binding affinity for FcγRIIIa (CD16), which resulted in enhanced antibody-dependent cellular cytotoxicity (ADCC) against FucGM1-expressing tumor cell lines. BMS-986012–mediated tumor cell killing was also observed in complement-dependent cytotoxicity (CDC) and antibody-dependent cellular phagocytosis (ADCP) assays. In several mouse SCLC models, BMS-986012 demonstrated efficacy and was well tolerated. In the DMS79 xenograft model, tumor regression was achieved with BMS-986012 doses of 0.3 mg/kg and greater; antitumor activity was enhanced when BMS-986012 was combined with standard-of-care cisplatin or etoposide. In a syngeneic model, tumors derived from a genetically engineered model of SCLC were treated with BMS-986012 or anti-FucGM1 with a mouse IgG2a Fc and their responses evaluated; when BMS-986012 was combined with anti–PD-1 or anti-CD137 antibody, therapeutic responses significantly improved. Conclusions: Single-agent BMS-986012 demonstrated robust antitumor activity, with the addition of chemotherapeutic or immunomodulatory agents further inhibiting SCLC growth in the same models. These preclinical data supported evaluation of BMS-986012 in a phase I clinical trial of patients with relapsed, refractory SCLC. Clin Cancer Res; 24(20); 5178–89. ©2018 AACR.

The microstructure and mechanical properties of the butt joint of 1100 MPa grade hot-rolled low-carbon steel by laser welding were investigated by scanning electron microscopy, micro-hardness and tensile tests. The yield strength and tensile strength of the laser welded joint reached 100.2 and 99.5% of the base material (BM), respectively. However, the elongation of the welded joint only reached about 60% of BM. The lowest and highest hardness areas both existed in the incomplete recrystallization zone. The width of the softened area of the welded joint is about 240–260 μm. The element distribution has no obvious change for C, Cr, Si, Mn, Ti, etc.

Many important processes for Seagrass ecology depend on leaf age. Through biweekly census, from April to November 2015, seasonal variation in leaf age structure of the Eelgrass Zostera marina L. was studied in Swan Lake of Shandong Peninsula, China by examining density, dimensions and biomass of shoots relative to leaf ages. In addition, we examined if variability in leaf age structure was related to water temperature and/or underwater irradiance. The leaf density, number of leaves per shoot, leaf area index, total leaf area per shoot and leaf biomass of young and old Z. marina leaves were far less than those of the mature leaves with percentages of >50% in all leaf ages. The leaf age structure of mature Z. marina leaves exhibited a clear seasonal variation with significant peaks in early July, which corresponded with the seasonal changes in shoot density and biomass of Z. marina. The results demonstrated that mature leaves would be a leading component in leaf age structure of Z. marina. Curve Estimation revealed that the seasonal variation in leaf age structure of Eelgrass was closely positively related to water temperature. The results provided data that could prove helpful in quantitative studies of leaf-age dependent aspects of Seagrass ecology.

The programmed death protein (PD-1) and its ligand (PD-L1) play critical roles in a checkpoint pathway cancer cells exploit to evade the immune system. A same-day PET imaging agent for measuring PD-L1 status in primary and metastatic lesions could be important for optimizing drug therapy. Herein, we have evaluated the tumor targeting of an anti-PD-L1 adnectin after 18F-fluorine labeling. Methods: An anti-PD-L1 adnectin was labeled with 18F in 2 steps. This synthesis featured fluorination of a novel prosthetic group, followed by a copper-free click conjugation to a modified adnectin to generate 18F-BMS-986192. 18F-BMS-986192 was evaluated in tumors using in vitro autoradiography and PET with mice bearing bilateral PD-L1-negative (PD-L1(-)) and PD-L1-positive (PD-L1(+)) subcutaneous tumors. 18F-BMS-986192 was evaluated for distribution, binding, and radiation dosimetry in a healthy cynomolgus monkey. Results:18F-BMS-986192 bound to human and cynomolgus PD-L1 with a dissociation constant of less than 35 pM, as measured by surface plasmon resonance. This adnectin was labeled with 18F to yield a PET radioligand for assessing PD-L1 expression in vivo. 18F-BMS-986192 bound to tumor tissues as a function of PD-L1 expression determined by immunohistochemistry. Radioligand binding was blocked in a dose-dependent manner. In vivo PET imaging clearly visualized PD-L1 expression in mice implanted with PD-L1(+), L2987 xenograft tumors. Two hours after dosing, a 3.5-fold-higher uptake (2.41 ± 0.29 vs. 0.82 ± 0.11 percentage injected dose per gram, P < 0.0001) was observed in L2987 than in control HT-29 (PD-L1(-)) tumors. Coadministration of 3 mg/kg ADX_5322_A02 anti-PD-L1 adnectin reduced tumor uptake at 2 h after injection by approximately 70%, whereas HT-29 uptake remained unchanged, demonstrating PD-L1-specific binding. Biodistribution in a nonhuman primate showed binding in the PD-L1-rich spleen, with rapid blood clearance through the kidneys and bladder. Binding in the PD-L1(+) spleen was reduced by coadministration of BMS-986192. Dosimetry estimates indicate that the kidney is the dose-limiting organ, with an estimated human absorbed dose of 2.20E-01 mSv/MBq. Conclusion:18F-BMS-986192 demonstrated the feasibility of noninvasively imaging the PD-L1 status of tumors by small-animal PET studies. Clinical studies with 18F-BMS-986192 are under way to measure PD-L1 expression in human tumors.

Electron–hole pair separation efficiency and adsorption performance of photocatalysts to CO2 are the two key factors affecting the performance of photocatalytic CO2 reduction with H2O. Distinct from conventional promoter addition, this study proposed a novel approach to address these two issues by tuning the own surface features of semiconductor photocatalyst. Three ZnO samples with different morphologies, surface area, and defect content were fabricated by varying preparation methods, characterized by XRD, TEM, and room-temperature PL spectra, and tested in photoreduction of CO2 with H2O. The results show that the as-prepared porous ZnO nanosheets exhibit a much higher activity for photoreduction of CO2 with H2O when compared to ZnO nanoparticles and nanorods attributed to the existence of more defect sites, that is, zinc and oxygen vacancies. These defects would lower the combination rate of electron–hole pair as well as promote the formation of basic zinc carbonate by Lewis acid–base interaction, which...

In the present study the extrusion process was performed on a novel biodegradable Mg-1.5Zn-0.6Zr-0.2Sc alloy, and the microstructure, mechanical properties, and in vitro degradation behavior of the alloy were investigated. The microstructure observation showed that the grain sizes were significantly refined after extrusion. However, there existed some long striped structures with relatively large grains along the extrusion direction. The strength, elongation, and hardness of the alloy were all improved after extrusion. The fracture mode transferred from quasi-cleavage fracture to ductile and cleavage mixed fracture. The strengthening mechanism belongs to grain refining strengthening. The cross section of the as-extruded Mg-1.5Zn-0.6Zr-0.2Sc alloy showed lower degradation rate and more uniform degradation mode compared with the longitudinal section. This can be explained by much refined grains, high microstructure uniformity, and low dislocation density.

Z3CN20-09M duplex steels are thermally aged at 400oC for up to 20,000 h. The mechanical properties have been characterized by Charpy V-notch impact test and nanoindentation test. It is found that the nanohardness in ferrite increases and the impact toughness decreases with aging time. Moreover, the distribution of alloying elements has been carefully characterized using atom probe tomography (APT). The results indicate that the ferrite decomposes into Cr-rich α' and Cr-lean α phase during the thermal aging and Ni-rich G-phase forms in ferrite. The effect of aging time on solute nanostructure has been investigated systematically.

Al 2 O 3 beads with various pore sizes were successfully synthesized by combining ammonium alginate assisted sol-gel and emulsion template methods using liquid paraffin as pore-enlarging agent and polysorbate 80 as dispersant. The great influences of textural structure of Al 2 O 3 beads on the activity and stability of PtSn-La/Al 2 O 3 catalyst for isobutane dehydrogenation were investigated by N 2 adsorption-desorption, XRD, NH 3 -TPD, SEM, HRTEM, H 2 -TPR, XPS, isobutene transient response experiment and TG analysis. The results showed that Al 2 O 3 beads with tunable pore sizes (7.4–13.3 nm), surface areas (294-322 m 2 /g) and pore volumes (0.56–1.1cm 3 /g) could be obtained by changing the addition amount of liquid oil. Moreover, the transient response experiment stated that the PtSn-La/Al 2 O 3 catalysts with large pore sizes could enhance the diffusion speed of the gas molecule in the channels. With increasing the amount of liquid oil from 0 to 20%wt, the particle sizes of platinum decreased from 2.86 nm to 1.45 nm, the amount of carbon deposit declined by 30.6% and the final yield of isobutene increased from 42% to 48%. The enhancement of catalytic performance of PtSn-La/Al catalyst with larger pore size was related to its smaller Pt particles, lower surface acidity and diffusion resistance of reaction products in the pore.

Background: Tumor-infiltrating CD4+ FOXP3+ regulatory T cells (Tregs) are a major driver of the immunosuppressive tumor microenvironment. CC chemokine receptor 8 (CCR8) expression is selectively upregulated in tumor-resident Tregs in multiple cancers, including breast, colon, and lung. These CCR8+ Tregs represents a highly activated and suppressive subset. High abundance of CCR8+ Tregs in these tumor types is associated with poor prognosis. Therefore, depletion of CCR8+ Tregs may be a promising approach to specifically address whether removal of Tregs will lead to augmentation of antitumor immunity. We evaluate the effect of CCR8-mediated Treg depletion alone and in combination with PD-1 blockade on antitumor immune responses in preclinical mouse tumor models and in a human tumor explant system. Methods: Human CCR8 and FOXP3 gene-gene correlations in The Cancer Genome Atlas (TCGA) were analyzed. Treg-specific protein expression of CCR8 was evaluated on tumor and peripheral tissues for both human and mice via flow cytometry and IHC. Single-cell RNA seq was performed on human tumors for differential gene expression analysis of CCR8+ Tregs. A variety of syngeneic mouse tumor models were used to determine antitumor activity of anti-CCR8 mouse surrogate antibody alone and in combination with anti–PD-1. Phenotyping of immune cell subsets was also conducted for proximal and distal pharmacodynamic changes. BMS-986340 (anti-CCR8 hIgG1-nonfucosylated [NF] antibody) was developed and used for CCR8+ Treg depletion in human tumor explants. Results: CCR8 gene expression had the highest correlation with FOXP3 in most cancer types in TCGA. CCR8 was specifically expressed on FOXP3hi Tregs while not on FOXP3mid and FOXP3neg effector T cells in patient tumors. Limited expression in peripheral blood, thymus, and skin resident T cells was seen. A similar expression profile was observed in mouse tumor models. CCR8 antibody–mediated Treg depletion and subsequent pro-inflammatory responses (↑CD8/CD4/IFNγ/GranzymeB/Ki67) induced robust tumor growth inhibition as a single agent in immunogenic tumor models and in combination with anti–PD-1 in I-O–resistant models. In these models, a dose-dependent pharmacokinetic/pharmacodynamic/efficacy relationship was observed. We also found tumor Treg–specific depletion in human tumor explant models upon treatment with BMS-986340. Conclusions: BMS-986340, a novel anti-CCR8 NF monoclonal antibody in development, led to measurable CCR8+ Treg depletion in human tumor explants. CCR8+ Treg depletion led to robust antitumor activity alone and in combination with anti–PD-1 in mouse tumor models. These results support further clinical evaluation of CCR8 depletion in combination with immune checkpoint blockade as a novel immunotherapy for cancer. Citation Format: Ruth Lan, Amy Jhatakia, Joseph Campbell, Nathan Siemers, Kai Lu, Mohammed Nasser, Tetiana Grigoriev, Priti Singh, Logan Vlach, Tim Sproul, Chengyue Zhang, Xi-Tao Wang, Olufemi Adelakun, Felix Findeisen, Pavel Strop, Arvind Rajpal, Natalie Bezman, Michael Quigley, Renu Jain. Highly selective anti-CCR8 antibody-mediated depletion of regulatory T cells leads to potent antitumor activity alone and in combination with anti-PD-1 in preclinical models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6694.

The hot tensile deformation behaviors of 316LN austenitic stainless steel (ASS) were studied on a Gleeble-1500D thermal simulator under the deformation temperature of 1173-1473 K and strain rate of 0.01-1 s-1. The effects of deformation temperature and strain rate on hot deformation behaviors were analyzed. Based on experimental data, the constitutive equation was established, and the predicted peak stresses by the developed model agree well with the experimental data. Microstructure near the fracture and the percentage reduction of area were studied, and the results showed that the microstructural evolution has great influences on the percentage reduction of area. Under the deformation temperature of 1473K with the strain rate of 1s-1, the grain was the finest and most homogenous, and in this deformation condition the percentage reduction of area was the highest of 79.8%.

In this study, the hot deformation experiments of 316LN stainless steel under various deformation temperature and various strain rates were conducted on a Gleeble thermal simulator. The true stain and true stress data were obtained and the Arrhenius-type constitutive model was developed, which can be used to accurately predict the flow stress of the studied steel under certain deformation conditions.

A novel phosphorus-doped porous carbon/graphene composite was adopted as electrode material of super-capacitor, which showed excellent electrochemical performance compared with carbon material without phosphorus heteroatom by means of cyclic voltammetry, the charge/discharge property, impedance characteristics, cycle life, and stability. The P-enriched carbons sample offered an outstanding capacitive behavior, which had specific capacitance 277 F/g and was able to withstand at a wide voltage window of 1.6 V with 90.8% performance retention after 10,000 cycles at a current density of 10 Ag&minus, 1, providing a higher energy density 26.42 Wh/kg. In addition, because the thermal effect in charge and discharge process can make the supercapacitor temperature rise rapidly in a short time and affect the electrical performance, temperature characteristic is one of the important characteristics to be considered in practical application. In this paper, a two-dimensional thermal model for commonly used coiling supercapacitor with p-doped porous carbon/graphene composite as electrode material was established, and the temperature distribution of supercapacitor and the variation of internal temperature under different conditions were analyzed by finite element method. The results show that the maximum temperature appears near the center, and the maximum temperature is related to the applied current and the number of cycles. With the increase of the current, the maximum internal temperature is increased sharply, and it is kept constant after the number of cycles reaches a certain value. Cooling measures should be taken when the maximum temperature exceeds the allowable temperature range.

Aluminum metal matrix composites (Al-MMCs) are new promising materials for aviation, aerospace and automotive industries. However, due to the poor weldability they have very limited applications. In this paper, the authors present the welding achievements of Al-MMCs developed by their scientific research team in recent years. Laser welding, liquid phase impact diffusion welding and vacuum brazing were utilized. Based on analysis of microstructure, good joints can be achieved by using these welding methods.

Dent's disease is a nephrolithiasis disorder associated with hypercalciuria and low molecular weight proteinuria that is caused by mutations in the voltage-gated chloride channel ClC-5. Because the exact cause of hypercalciuria in this disease is unknown and could come from a renal, intestinal, or bone origin, we have investigated overall calcium handling in the ClC-5 knockout mouse (ClC-5 KO). On a high calcium diet, ClC-5 KO mice had elevated serum 1alpha,25-dihydroxyvitamin D3 (1alpha,25D3), alkaline phosphatase (AP), osteocalcin (OC), and urinary deoxypyridinoline (DPD), but serum parathyroid hormone (PTH), calcium, and intestinal calcium uptake was similar to that of wild-type (WT) mice. A 30-fold decrease in dietary calcium intake caused elevation of serum PTH and urinary cyclic adenosine monophosphate in ClC-5 KO mice and decreased the renal calcium excretion, which still remained 2-fold above that of WT mice. On this low calcium diet, both groups of mice had the same serum 1alpha,25D3, with similar increments in intestinal calcium absorption, serum AP, OC, and urinary DPD. These data indicate that the hypercalciuria in the ClC-5 KO mice on low and high calcium diets is of bone and renal origin and is not caused by increased intestinal calcium absorption, despite an elevated serum 1alpha,25D3. These mice data suggest that young patients with this disease may have a propensity for altered bone homeostasis that should be monitored clinically.

To compare the predictive values of eight staging systems for primary liver cancer in the prognosis of combined hepatocellular-cholangiocellular carcinoma (cHCC-CC) patients after surgery.The clinical data of 54 cHCC-CC patients who underwent hepatectomy or liver transplantation from May 2005 to Augest 2013 in Chinese PLA General Hospital were collected. We evaluated the prognostic value of the Okuda staging system, Cancer of the Liver Italian Program (CLIP) score, French staging system, Barcelona Clinic Liver Cancer (BCLC) staging system, 7th edition of tumour-node-metastasis (TNM) staging system for hepatocellular carcinoma and intrahepatic cholangiocarcinoma (ICC), Japan Integrated Staging (JIS) score, and Chinese University Prognostic Index. The distribution, Kaplan-Meier method, Log-rank test, and area under a receiver operating characteristic curve were used to compare the prognosis-predicting ability of these different staging systems in 54 cHCC-CC patients after surgery.The TNM staging system for ICC and JIS score had a better distribution of cases. The 12-and 24-month survivals of the entire cohort were 65.5% and 56.3%, respectively. A Log-rank test showed that there was a significant difference existing in the cumulative survival rates of different stage patients when using TNM staging system for ICC (stage 1 vs. stage 2, P=0.012; stage 2 vs. stage 3-4, P=0.002), Okuda staging system (stage 1 vs. stage 2, P=0.025), and French staging system (stage A and stage B, P=0.045). The 12-and 24-month area under curve of TNM staging system for ICC, BCLC staging system, JIS score, and CLIP score were 0.836 and 0.847, 0.744 and 0.780, 0.723 and 0.764, and 0.710 and 0.786, respectively.The 7th edition of TNM staging system for ICC has superior prognostic value to other seven staging systems in cHCC-CC patients undergoing surgical treatment.

Acute liver failure (ALF) is a reversible disorder that is associated with an abrupt loss of hepatic mass, rapidly progressive encephalopathy and devastating complications. Despite its high mortality, an emergency liver transplantation nowadays forms an integral part in ALF management and has substantially improved the outcomes of ALF. Here, we report the case of a 32-year-old female patient who was admitted with grade IV hepatic encephalopathy (coma) following drug-induced ALF. We performed an emergency auxiliary partial orthotopic liver transplantation with a "high risk" graft (liver macrovesicular steatosis approximately 40%) from a living donor. The patient was discharged on postoperative day 57 with normal liver function. Weaning from immunosuppression was achieved 9 mo after transplantation. A follow-up using CT scan showed a remarkable increase in native liver volume and gradual loss of the graft. More than 6 years after the transplantation, the female now has a 4-year-old child and has returned to work full-time without any neurological sequelae.

A series of ZnO/ZnS core/shell nanorods with different ZnS/ZnO molar ratios was synthesized via a new water bath route. The nanorods have a diameter of about 100 nm and a length ranging from a few hundred nanometers to several micrometers. They are formed by coating ZnO nanorod with a layer of porous ZnS shell mainly consisting of crystals which are about 12 nm in diameter. The results showed that the deposition thickness of the ZnS shell layer strongly affected the morphologies, surface area, structure, photo absorption and photocatalytic performance of the ZnO/ZnS core/shell nanorods. The as-prepared ZnO/ZnS core/shell nanorods exhibited a higher photocatalytic activity for H2 evolution from the glycerol/water mixtures compared with the ZnO nanorods under the same conditions. The maximum H2 production was 2608.7 and 388.4 μmol h−1 gcat−1 under UV and solar-simulated light irradiation and the corresponding quantum efficiencies were 22% and 13%, respectively. The deposition thickness of the ZnS shell and the interaction between the ZnO rod and ZnS shell and the core/shell structure with n-p heterojunction substantially influence the optical and catalytic performance of the ZnO/ZnS core/shell nanorods.

Introduction: Anti-tumor immune response is controlled by a complex interaction between the immune system, tumor cells, and associated stroma. Expression of major histocompatibility complex class I (MHC I) and MHC class II (MHC II) antigens may be dysregulated in cancer, leading to alterations in the tumor antigen presentation profile. Stage I–III resected NSCLC tumor samples were profiled to study the relationship between MHC I, MHC II, and programmed death ligand 1 (PD-L1) tumor cell (TC) expression with tumor-associated inflammation. Here we report the potential impact of respective profiles on tumor immune response. Methods: 53 adenocarcinoma (NSCLC-AD) and 51 squamous cell carcinoma (NSCLC-SQ) stage I–III resected formalin-fixed, paraffin-embedded tumor specimens from commercial sources were stained by immunohistochemistry for MHC I (HLA-A,B,C), MHC II (HLA-DP,DQ,DR), and PD-L1 and TC expression was assessed by manual pathologist review. CD8+ cell density was quantified using Definiens image analysis algorithms for the intraepithelial tumor region. RNA extracted from the samples was analyzed by RNAseq, with data available for 48 NSCLC-AD and 46 NSCLC-SQ specimens. A 25-gene IFN-gamma gene signature (IFNG score) was used to study the association of T-cell inflammation with other biomarkers. Results: Of all NSCLC-AD and NSCLC-SQ specimens, 85% showed either complete (1%; MHCIIhi) in 60% of NSCLC-AD and 18% of NSCLC-SQ. Quantitative analysis of CD8 in the tumor microenvironment (TME) revealed a significantly reduced intraepithelial density of CD8+ tumor-infiltrating lymphocytes (TILs) in NSCLC-AD (P=0.009) and NSCLC-SQ (P=0.01) with complete MHC I loss. In contrast, intraepithelial CD8+ TILs were significantly increased in MHCIIhi tumors for both NSCLC-AD (P=0.004) and NSCLC-SQ (P=0.006). Tumors displaying both MHCIIhi and retained MHC I demonstrated the highest TIL density. Gene expression associated with IFN-gamma response was increased in tumors with retained MHC I (P=0.0036) and MHCIIhi samples (P Conclusions: Understanding the role of the antigen presentation machinery in immune activation and evasion in the TME is important to predict responses to immunotherapy. These data suggest that the expression of MHC I, MHC II, and PD-L1 by tumor cells may play a role in guiding the localization of CD8+ T cells in the TME with differential effects dependent on histologic subtype. The lack of correlation between inflammation and PD-L1 expression in stage I–III NSCLC-SQ suggests an alternate PD-L1 induction mechanism in a subset of those tumors. Citation Format: Cyrus Hedvat, Keyur Desai, Dimple Pandya, Peter Szabo, Johannes Zimmermann, Jan Lesniak, Scott Ely, Sujaya Srinivasan, Xi-Tao Wang, Michele French, Robin Edwards. Tumor intrinsic properties associate with differential effects on CD8+ tumor-infiltrating lymphocyte density and immune gene expression in non-small cell lung cancer (NSCLC) samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1024.

Background: Integrated analysis of multi-omic data is important in understanding oncogenesis, pathology, and drug mechanism of action. Formalin-fixed, paraffin-embedded (FFPE) tissues comprise the bulk of archival specimens in hospitals and clinical trials. Commercial kits are commonly used to extract RNA, DNA, or protein for biomarker studies; however, high-quality extractions are challenging due to crosslinking. We explored the feasibility of performing proteomic, transcriptomic, and genetic analysis from limited FFPE samples collected in clinical trials through a pilot study in colorectal cancer (CRC) and normal tissue. Methods: Qiagen kits were employed, with some modifications, to extract nucleic acid and protein from FFPE sample lysates that would be amenable to next-generation sequencing and liquid chromatography/mass-spectrometry (LC/MS) proteomic profiling. Commercially sourced FFPE slides from CRC biopsies and normal colon, heart, and skin samples were processed (n = 10 each; ~600-mm2 by 5-μm/slice). RNA-seq library preparation was performed using the Illumina® TruSeq Access kit. Label-free LC/MS proteomic analysis was carried out using trypsin/Lys-C-digested protein fragments on an EASY-nLC™ 1200 coupled to a Q Exactive™ Plus (Thermo Fisher). MS data were processed with MaxQuant to estimate protein abundance (Cox and Mann, Nat Biotech 2008). Results: Of 40 samples tested, 38 yielded quantifiable nucleic acid and protein of sufficient quality for transcriptional and proteomic analyses. Mean RNA yield was ~300 ng (150 ng-1.2 µg). RNA degradation was significant, with a mean DV>200 of 45% (Agilent Bioanalyzer). Mean DNA yield was 530 ng, with a mean fragment size of 2 kb. Mean protein yield was 50 µg (1 µg-300 µg). RNA libraries had a mapping rate range of 85%-90% and a coding rate range of 70%-80%; ~16,000 genes were reliable detected (log2 TPM > 1). The number of unique proteins detected ranged from 3,000 to 4,000 for CRC, normal colon, and heart samples; skin samples averaged 1,000 proteins and had the lowest overall yield. Correlation of RNA and protein expression for the same genes was weak (Spearman's rho ~0.3-0.4) at the per-sample level but statistically significant. Using linear models, we identified differentially expressed transcripts and proteins between the CRC and normal colon samples. Pathway enrichment analysis of both modalities indicated changes in cell cycle, which is consistent with rapid growth of tumor cells. Changes in nucleoside metabolism, extracellular matrix remodeling, and innate immune response were more apparent at the protein level. Conclusion: We found that multi-omic analysis was feasible with FFPE samples, and proteomics can be used to validate RNA results. Additionally, proteomics reveal post-translational events, such as extracellular matrix remodeling, that provide unique insights into cancer pathology. Citation Format: Vishal Patel, Ji Gao, Mingyi Liu, Aiqing He, Xi-Tao Wang, Kathryn Vanderlaag, Ariella Sasson, Stefan Kirov, Sunil Kuppasani, Omar J. Jabado, Kandasamy Ravi, Ashok Dongre, Julie Carman, Heidi LeBlanc. Integrated analysis of colorectal carcinoma by co-extraction of RNA, DNA and protein from FFPE tumor samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2707.

Thermo-physical properties of diamond reinforced Al composites were investigated. Volume fraction of diamond particles was up to 55%. In order to improve the interfacial bonding between diamond and aluminum, diamond particles were pre-coated with titanium using molten salt method. XRD and SEM observation showed that the Ti coating on diamond consists of carbide layer and metal layer, which mainly depend on temperature and time. The influences of the Ti coating on interfacial characteristic and the thermo-physical properties of the composites were studied. The interfacial characterization and thermal diffusivity measurements indicated that Ti coated diamond was more favorable on interfacial bonding and thermal properties. Ti coating on diamond resulted in an increase of thermal conductivity of the composites, from 200 to 430 W/mK along with a coefficient of thermal expansion of 6.40 × 10-6/K.

The rheo-diecasting (RDC) process, a novel semi-solid processing technology, was used to produce cast components with high integrity, fine and uniform microstructure, and therefore enhanced performance. AZ91D samples with 1-3 mass%Gd were solidified by RDC process. It was found that under intensive forced convection, the primary α-Mg phase produced inside the twin-screw slurry maker had fine particle size, spherical morphology and uniform distribution throughout the samples. Microstructure observations showed that Gd addition resulted in the formation of Al2Gd which dispersed in the α-Mg matrix. The size and amount of β-Mg17Al12 phase was reduced and its continuity was broken, which was the main reason for improving mechanical properties of the AZ91D alloy at high temperatures. The amount of Al2Gd particles increased with increasing Gd addition. From EPMA quantitative analysis, almost all Gd reacted with Al, leading to the low concentration of Gd in the α-Mg matrix. The Vickers hardness increased monotonously from HV=50.4 to HV=67.3 with increasing amount of Gd. This improvement was attributed to the consummation of aluminum in melt by precipitation of the Al2Gd phases.

A novel thermomechanical process to manufacture hot-rolled transformation-induced plasticity (TRIP) steels was developed based on dynamic transformation of undercooled austenite (DTUA). Between DTUA and the isothermal bainitic treatment, only one-step controlled-cooling was required. The microstructure evolution of hot-rolled C-Mn-Si and C-Mn-Al-Si TRIP steels based on DTUA was investigated by hot uniaxial compression tests using a Gleeble1500 simulation test machine. The results indicated that during DTUA, the kinetics of ferrite formation was fast, the volume fraction of ferrite formed was determined by applied strain. In comparison with the process based on static transformation of austenite, a more uniform multiphase microstructure with fine ferrite grains was formed, the bainite packets were small and had relatively random orientations, the retained austenite distributed uniformly and had relatively high volume fraction. Hot-rolled TRIP steels based on DTUA demonstrated better mechanical properties, especially for C-Mn-Al-Si TRIP steel.

In order to determine the ductile to brittle transition behavior of nuclear primary pipe material (Z3CN20.09M) during the thermal aging procedure, instrumented impact tests at different temperatures were performed on Z3CN20.09M aged at 400°C for up to 3000 hours. The load-deflection curves from the instrumented impact tests described both the loading and the fracture stages, from which the dynamic strengths and energy were calculated. The results indicated that the thermal aging decreases the impact energy and shifts the ductile-to-brittle transition curves to higher temperatures.

In this studies, Mg(Al)-50 vol.% SiC particle composites were fabricated by pressure infiltration with the two different matrix base materials of Mg and Mg-6Al alloy. Thermal conductivities of the two composites were compared and the effect of addition was studied using X-ray diffraction and scanning electron microscopy (SEM). The analysis of the X-ray diffraction shows that the Al-addition to the Mg causes a contraction of the Mg lattice parameters, which brings a better matching between the matrix and SiC particle. The microstructure and the fracture surface of the composite were characterized by using SEM. The alloying element Al exists mainly in the form of α solid solution, which has a uniform distribution in matrix. From calculation, the interfacial thermal barrier resistance of the Mg-6Al/SiCp composite is about one order of magnitude lower than that of the Mg/SiCp composite.

A physical model for austenite recrystallization of steel concerning TMCP is developed. Dislocation density plays a key role as recrystallization driving force. The dislocation density change is a result of competition between dislocation generation and dynamic recovery. Recrystallization is described as a nucleation-growth process. An abnormal subgrain growth mechanism is introduced for nucleation. A few subgrains fulfilling abnormal growth conditions will stand out and become nuclei of recrystallization. The recrystallized grain grows to the deformed materials driven by the stored energy. Oswald ripening occurs for grains surrounded by recrystallized grains. The models were verified by laboratory simulation results for selected austenite stainless steels. It showed good agreement between predicted and experimental results.

Susceptibility of hydrogen embrittlement of a super grade AISI 420 tool steel was studied. Tensile samples were cathodically charged to different hydrogen level. Hydrogen induced mechanical property degradation was measured by tensile tests at a low strain rate. Fractography of broken surfaces was observed using SEM. Relationship between hydrogen content and tensile strength and elongation were studied. Critical hydrogen contents were obtained for different heat treatment states. It was found that for annealed materials could stand for a 3.5ppm hydrogen for keeping 80% of original ductility, and the effect of hydrogen on strength was unobvious. However, for material quenched and tempered at 250°C, only 0.3ppm hydrogen could lead the ductility drop to 80% of original. The material quenched and tempered at 500°C was more sensitive on hydrogen, less than 1ppm hydrogen could lead the strength drop to 80% of original.

Pure laparoscopic liver resection (PLLR) has been reported to be as safe and effective as open liver resection (OLR) for liver lesions, and it is associated with less intraoperative blood loss, shorter hospital stay, and lower complication rate. However, studies comparing PLLR with OLR in elderly patients were limited. The aim of this study was to analyze the short-term outcome of PLLR versus OLR for primary liver carcinoma (PLC) in elderly patients. Between January 2008 and October 2014, 30 consecutive elderly patients (≥70 years) who underwent PLLR for PLC were included into analysis. Sixty patients who received OLR for PLC during the same study period were also included as a case-matched control group. Patients were well matched in terms of age, sex, comorbid illness, Child Pugh class, American Society of Anesthesiologists grade, tumor size, tumor location, and extent of hepatectomy. No significant differences were observed with regard to patient preoperative baseline status, median tumor size (Group PLLR 4.0 cm vs Group OLR 5.0 cm, P = 0.125), tumor location, extent of hepatectomy, and operation time (Group PLLR 133 minutes vs Group OLR 170 minutes, P = 0.073). Compared with OLR, the PLLR group displayed a significantly less frequent Pringle maneuver application (10.0% vs 70.0%, P

Purpose: The oncofetal antigen, human chorionic gonadotropin β subunit (hCGβ), is expressed by a number of carcinomas and is a prognostic indicator in renal, colorectal, bladder, and pancreatic cancers. We describe the development of a novel antibody-based dendritic cell (DC)-targeted cancer vaccine capable of eliciting cellular immune responses directed against hCGβ.Experimental Design: The tumor-associated antigen hCGβ was coupled genetically to a human anti-DC antibody (B11). The resulting fusion protein (B11-hCGβ) was evaluated for its ability to promote tumor antigen-specific cellular immune responses in a human in vitro model. Monocyte-derived human DCs from normal donors were exposed to purified B11-hCGβ, activated with CD40 ligand, mixed with autologous lymphocytes, and tested for their ability to promote hCGβ-specific proliferative and cytotoxic T-lymphocyte responses.Results: B11-hCGβ was found to be a soluble, well-defined, and readily purified product that specifically recognized the human mannose receptor via the B11 antibody portion of the fusion protein. B11-hCGβ functionally promoted the uptake and processing of tumor antigen by DCs, which led to the generation of tumor-specific HLA class I and class II-restricted T-cell responses, including CTLs capable of killing human cancer cell lines expressing hCGβ.Conclusions: Although other hCG vaccines have been shown to be capable of eliciting antibody responses to hCGβ, this is the first time that cellular immune responses to hCGβ have been induced by a vaccine in a human system. This DC-targeted hCGβ vaccine holds promise for the management of a number of cancers and merits additional clinical development.

The most common defect in cystic fibrosis, deletion of phenylalanine from position 508 of the cystic fibrosis transmembrane conductance regulator (Delta F508 CFTR), decreases the trafficking of this protein to the cell surface membrane. Previous studies have shown that low temperature and high concentrations of glycerol or trimethylamine N-oxide can partially counteract the processing defect of Delta F508 CFTR. The present study investigates whether physiologically relevant concentrations of organic solutes, accumulated by cotransporter proteins, can rescue the misprocessing of Delta F508 CFTR. Myoinositol alone or myoinositol, betaine, and taurine given sequentially increased the processing of core-glycosylated, endoplasmic reticulum-arrested Delta F508 CFTR into the fully glycosylated form of CFTR in IB3 cells or NIH 3T3 cells stably expressing Delta F508 CFTR. Pulse-chase experiments using transiently transfected COS7 cells demonstrated that organic solutes also increased the processing of the core-glycosylated form of green fluorescent protein-Delta F508 CFTR. Moreover, the prolonged half-life of the complex-glycosylated form of GFP-Delta F508 CFTR suggests that this treatment stabilized the mature form of the protein. In vitro studies of purified NBD1 stability and aggregation showed that myoinositol stabilized both the Delta F508 and wild type CFTR and inhibited Delta F508 misfolding. Most significantly, treatment of CF bronchial airway cells with these transportable organic solutes restores cAMP-stimulated single channel activity of both CFTR and outwardly rectifying chloride channel in the cell surface membrane and also restores a forskolin-stimulated macroscopic 36Cl- efflux. We conclude that organic solutes can repair CFTR functions by enhancing the processing of Delta F508 CFTR to the plasma membrane by stabilizing the complex-glycosylated form of Delta F508 CFTR.