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Table S1. Inter-tumor heterogeneity of T-cell densities in tumor microenvironment in dMMR vs pMMR stage III colon cancers

Figure S1 Study flow of N0147 patient tumor samples Figure S2 Heterogeneity of lymphocytic infiltrate between patients in mismatch repair deficient (dMMR) and proficient (pMMR) tumors. The distribution of T-cell densities (score 0-100) are shown as follows: (a) CD3+ in the core of the tumor, (b) CD8+ at the invasive margin, (c) CD8+ in the core of the tumor. For each T-cell subtype, density of tumor infiltration was higher in dMMR vs pMMR tumors (each p

Table S1: Baseline patient demographics and disease characteristics. Table S2: mRNA/IHC AREG/EREG concordance table. Figure S1: AREG/EREG IHC scatterplot. Table S3: Prognostic effect of dichotomous AREG/EREG. Table S4: AREG/EREG and RECIST response rate. Table S5: AREG and EREG as individual continuous variables. Table S6: AREG/EREG predictive analyses adjusted for BRAF mutation, PTL and peritoneal metastases. Table S7: Exploratory analyses of different AREG/EREG cut points. Table S8: AREG/EREG IHC in biopsy and resection specimens.

Purpose:High tumor mRNA levels of the EGFR ligands amphiregulin (AREG) and epiregulin (EREG) are associated with anti-EGFR agent response in metastatic colorectal cancer (mCRC). However, ligand RNA assays have not been adopted into routine practice due to issues with analytic precision and practicality. We investigated whether AREG/EREG IHC could predict benefit from the anti-EGFR agent panitumumab.Experimental Design:Artificial intelligence algorithms were developed to assess AREG/EREG IHC in 274 patients from the PICCOLO trial of irinotecan with or without panitumumab (Ir vs. IrPan) in RAS wild-type mCRC. The primary endpoint was progression-free survival (PFS). Secondary endpoints were RECIST response rate (RR) and overall survival (OS). Models were repeated adjusting separately for BRAF mutation status and primary tumor location (PTL).Results:High ligand expression was associated with significant PFS benefit from IrPan compared with Ir [8.0 vs. 3.2 months; HR, 0.54; 95% confidence interval (CI), 0.37–0.79; P = 0.001]; whereas low ligand expression was not (3.4 vs. 4.4 months; HR, 1.05; 95% CI, 0.74–1.49; P = 0.78). The ligand-treatment interaction was significant (Pinteraction = 0.02) and remained significant after adjustment for BRAF-mutation status and PTL. Likewise, RECIST RR was significantly improved in patients with high ligand expression (IrPan vs. Ir: 48% vs. 6%; P < 0.0001) but not those with low ligand expression (25% vs. 14%; P = 0.10; Pinteraction = 0.01). The effect on OS was similar but not statistically significant.Conclusions:AREG/EREG IHC identified patients who benefitted from the addition of panitumumab to irinotecan chemotherapy. IHC is a practicable assay that may be of use in routine practice.

Purpose:Colorectal cancers with deficient DNA mismatch repair (dMMR) are presumed to uniformly have dense lymphocytic infiltration that underlies their favorable prognosis and is critical to their responsiveness to immunotherapy, as compared with MMR-proficient (pMMR) tumors. We examined T-cell densities and their potential heterogeneity in a large cohort of dMMR tumors.Experimental Design:CD3+ and CD8+ T-cell densities were quantified at the invasive margin (IM) and tumor core (CT) in 561 stage III colon cancers (dMMR, n = 278; pMMR, n = 283) from a phase III adjuvant trial (N0147). Their association with overall survival (OS) was determined using multivariable Cox analysis.Results:Although CD3+ and CD8+ T-cell densities in the tumor microenvironment were higher in dMMR versus pMMR tumors overall, intertumoral heterogeneity in densities between tumors was significantly higher by 30% to 88% among dMMR versus pMMR cancers (P < 0.0001 for all four T-cell subtypes [CD3+IM, CD3+CT, CD8+IM, CD8+CT]). A substantial proportion of dMMR tumors (26% to 35% depending on the T-cell subtype) exhibited T-cell densities as low as that in the bottom half of pMMR tumors. All four T-cell subtypes were prognostic in dMMR with CD3+IM being the most strongly prognostic. Low (vs. high) CD3+IM was independently associated with poorer OS among dMMR (HR, 4.76; 95% confidence interval, 1.43–15.87; P = 0.0019) and pMMR tumors (P = 0.0103).Conclusions:Tumor-infiltrating T-cell densities exhibited greater intertumoral heterogeneity among dMMR than pMMR colon cancers, with CD3+IM providing robust stratification of both dMMR and pMMR tumors for prognosis. Potentially, lower T-cell densities among dMMR tumors may contribute to immunotherapy resistance.

Purpose: High tumor mRNA levels of the EGFR ligands amphiregulin (AREG) and epiregulin (EREG) are associated with anti-EGFR agent response in metastatic colorectal cancer (mCRC). However, ligand RNA assays have not been adopted into routine practice due to issues with analytic precision and practicality. We investigated whether AREG/EREG IHC could predict benefit from the anti-EGFR agent panitumumab. Experimental Design: Artificial intelligence algorithms were developed to assess AREG/EREG IHC in 274 patients from the PICCOLO trial of irinotecan with or without panitumumab (Ir vs. IrPan) in RAS wild-type mCRC. The primary endpoint was progression-free survival (PFS). Secondary endpoints were RECIST response rate (RR) and overall survival (OS). Models were repeated adjusting separately for BRAF mutation status and primary tumor location (PTL). Results: High ligand expression was associated with significant PFS benefit from IrPan compared with Ir [8.0 vs. 3.2 months; HR, 0.54; 95% confidence interval (CI), 0.37–0.79; P = 0.001]; whereas low ligand expression was not (3.4 vs. 4.4 months; HR, 1.05; 95% CI, 0.74–1.49; P = 0.78). The ligand-treatment interaction was significant (Pinteraction = 0.02) and remained significant after adjustment for BRAF-mutation status and PTL. Likewise, RECIST RR was significantly improved in patients with high ligand expression (IrPan vs. Ir: 48% vs. 6%; P < 0.0001) but not those with low ligand expression (25% vs. 14%; P = 0.10; Pinteraction = 0.01). The effect on OS was similar but not statistically significant. Conclusions: AREG/EREG IHC identified patients who benefitted from the addition of panitumumab to irinotecan chemotherapy. IHC is a practicable assay that may be of use in routine practice.

Context.-The ability to determine ROS1 status has become mandatory for patients with lung adenocarcinoma, as many global authorities have approved crizotinib for patients with ROS1-positive lung adenocarcinoma. Objective.-To present analytical correlation of the VENTANA ROS1 (SP384) Rabbit Monoclonal Primary Antibody (ROS1 [SP384] antibody) with ROS1 fluorescence in situ hybridization (FISH). Design.-The immunohistochemistry (IHC) and FISH analytical comparison was assessed by using 122 non-small cell lung cancer samples that had both FISH (46 positive and 76 negative cases) and IHC staining results available. In addition, reverse transcription-polymerase chain reaction (RT-PCR) as well as DNA and RNA next-generation sequencing (NGS) were used to further examine the ROS1 status in cases that were discrepant between FISH and IHC, based on staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells considered as IHC positive. Here, we define the consensus status as the most frequent result across the 5 different methods (IHC, FISH, RT-PCR, RNA NGS, and DNA NGS) we used to determine ROS1 status in these cases. Results.-Of the IHC scoring methods examined, staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells considered as IHC positive had the highest correlation with a FISH-positive status, reaching a positive percentage agreement of 97.8% and negative percentage agreement of 89.5%. A positive percentage agreement (100%) and negative percentage agreement (92.0%) was reached by comparing ROS1 (SP384) using a cutoff for staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells to the consensus status. Conclusions.-Herein, we present a standardized staining protocol for ROS1 (SP384) and data that support the high correlation between ROS1 status and ROS1 (SP384) antibody. © 2020 College of American Pathologists. All rights reserved. Boehringer Ingelheim; Bristol-Myers Squibb; Pfizer; Novartis; Roche; AbbVie; Merck KGaA Dr Huang, Mr Smith, Dr Menzl, Dr Le, Mr Liu, Dr Ordinario, Dr Manohar, Mr Lee, Mr Rajamani, Mr Truong, Dr Jing Li, Ms Choi, Dr Jingchuan Li, Dr Pati, Dr Hanlon Newell, and Mr Pate are Roche employees and receive a salary from Roche. Dr Bubendorf received research support from Roche and received honoraria related to advisory board meetings (Pfizer, Roche). Dr Buettner is a cofounder and chief scientific officer of Targos Mol. Pathol. Inc, Kassel, Germany. Dr Kerr has received honoraria for advisory work and lectures from Ventana Medical Systems, Roche, and Pfizer; these payments were outside the scope of this work. Dr Lopez-Rios received funding and honoraria from Roche, Pfizer, and Thermo Fischer. Dr Marchetti has received honoraria for advisory work and lectures from Ventana Medical Systems, Roche, and Pfizer; these payments were outside the scope of this work. Dr Marondel is an employee and receives a salary from Pfizer. Dr Nicholson serves as a consultant and receives a fee from Merck, Boehringer Ingelheim, Pfizer (also research grant), Novartis, Astra Zeneca (also payment for lecture), Bristol-Myers Squibb, Roche, and ABBVIE. Dr Pauwels received research grants from Pfizer, Roche, and Astra Zeneca and was paid for attending advisory boards of BMS, MSD Astra Zeneca, Takedo, Roche, and Boehringer. Dr Thunnissen participates in Ventana advisory board meetings. The other authors (Dr Öz, Dr Penault-Llorca, and Dr Rossi) have no relevant financial interest in the products or companies described in this article.

Technologies such as external sensors, data acquisition, trend analysis, and cloud services make remote operation and maintenance of equipment status monitoring more convenient and lower maintenance costs. In this paper, an industrial robot operation and maintenance system is designed. First, the online monitoring of the robot is realized by collecting the control parameters, state parameters, fault information and external sensor monitoring information of the industrial robot. Then, the health baseline of the industrial robot is established based on the historical operation data. The online data is compared with the baseline data to realize the online assessment of the robot’s health status. Finally, a visualization platform for remote operation and maintenance of the robot is built to realize the online display of parameters, health trend tracking and fault status early warning, which is convenient for maintenance personnel to take timely countermeasures.

Purpose: Colorectal cancers with deficient DNA mismatch repair (dMMR) are presumed to uniformly have dense lymphocytic infiltration that underlies their favorable prognosis and is critical to their responsiveness to immunotherapy, as compared with MMR-proficient (pMMR) tumors. We examined T-cell densities and their potential heterogeneity in a large cohort of dMMR tumors. Experimental Design: CD3+ and CD8+ T-cell densities were quantified at the invasive margin (IM) and tumor core (CT) in 561 stage III colon cancers (dMMR, n = 278; pMMR, n = 283) from a phase III adjuvant trial (N0147). Their association with overall survival (OS) was determined using multivariable Cox analysis. Results: Although CD3+ and CD8+ T-cell densities in the tumor microenvironment were higher in dMMR versus pMMR tumors overall, intertumoral heterogeneity in densities between tumors was significantly higher by 30% to 88% among dMMR versus pMMR cancers (P < 0.0001 for all four T-cell subtypes [CD3+IM, CD3+CT, CD8+IM, CD8+CT]). A substantial proportion of dMMR tumors (26% to 35% depending on the T-cell subtype) exhibited T-cell densities as low as that in the bottom half of pMMR tumors. All four T-cell subtypes were prognostic in dMMR with CD3+IM being the most strongly prognostic. Low (vs. high) CD3+IM was independently associated with poorer OS among dMMR (HR, 4.76; 95% confidence interval, 1.43–15.87; P = 0.0019) and pMMR tumors (P = 0.0103). Conclusions: Tumor-infiltrating T-cell densities exhibited greater intertumoral heterogeneity among dMMR than pMMR colon cancers, with CD3+IM providing robust stratification of both dMMR and pMMR tumors for prognosis. Potentially, lower T-cell densities among dMMR tumors may contribute to immunotherapy resistance.

High tumor mRNA levels of the EGFR ligands amphiregulin (AREG) and epiregulin (EREG) are associated with anti-EGFR agent response in metastatic colorectal cancer (mCRC). However, ligand RNA assays have not been adopted into routine practice due to issues with analytic precision and practicality. We investigated whether AREG/EREG IHC could predict benefit from the anti-EGFR agent panitumumab.Artificial intelligence algorithms were developed to assess AREG/EREG IHC in 274 patients from the PICCOLO trial of irinotecan with or without panitumumab (Ir vs. IrPan) inHigh ligand expression was associated with significant PFS benefit from IrPan compared with Ir [8.0 vs. 3.2 months; HR, 0.54; 95% confidence interval (CI), 0.37-0.79;AREG/EREG IHC identified patients who benefitted from the addition of panitumumab to irinotecan chemotherapy. IHC is a practicable assay that may be of use in routine practice.

The ability to determineTo present analytical correlation of the VENTANA ROS1 (SP384) Rabbit Monoclonal Primary Antibody (ROS1 [SP384] antibody) withThe immunohistochemistry (IHC) and FISH analytical comparison was assessed by using 122 non-small cell lung cancer samples that had both FISH (46 positive and 76 negative cases) and IHC staining results available. In addition, reverse transcription-polymerase chain reaction (RT-PCR) as well as DNA and RNA next-generation sequencing (NGS) were used to further examine the ROS1 status in cases that were discrepant between FISH and IHC, based on staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells considered as IHC positive. Here, we define the consensus status as the most frequent result across the 5 different methods (IHC, FISH, RT-PCR, RNA NGS, and DNA NGS) we used to determine ROS1 status in these cases.Of the IHC scoring methods examined, staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells considered as IHC positive had the highest correlation with a FISH-positive status, reaching a positive percentage agreement of 97.8% and negative percentage agreement of 89.5%. A positive percentage agreement (100%) and negative percentage agreement (92.0%) was reached by comparing ROS1 (SP384) using a cutoff for staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells to the consensus status.Herein, we present a standardized staining protocol for ROS1 (SP384) and data that support the high correlation between ROS1 status and ROS1 (SP384) antibody.

BACKGROUND: Colorectal cancers with deficient DNA mismatch repair (dMMR) are presumed to uniformly have dense lymphocytic infiltration that underlies their favorable prognosis and is critical to their responsiveness to immunotherapy, as compared to MMR-proficient (pMMR) tumors. We examined T-cell densities and their potential heterogeneity in a large cohort of dMMR tumors. EXPERIMENTAL DESIGN: CD3(+) and CD8(+) T-cell densities were quantified at the invasive margin (IM) and tumor core (CT) in 561 stage III colon cancers (dMMR, n=278; pMMR, n=283) from a phase 3 adjuvant trial (N0147). Their association with overall survival (OS) was determined using multivariable Cox analysis. RESULTS: While CD3(+) and CD8(+) T-cell densities in the tumor microenvironment were higher in dMMR vs pMMR tumors overall, inter-tumoral heterogeneity in densities between tumors was significantly higher by 30–88% among dMMR vs pMMR cancers (P

The functional relationship and cross-regulation between damage-associated molecular patterns and NF‑κB in the tumor microenvironment remains unclear. In the present study, high-mobility group protein B1 (HMGB1) was secreted in response to feed second phase of NF‑κB activation from heat shock protein (HSP) 70 that may result in a higher invasion potential of hepatocarcinoma cells. HSP70 promoted the proliferation of H22 hepatocarcinoma cells through Toll-like receptor (TLR) 2 and TLR4 signaling and induced the early phosphorylation of NF-κB, which reached maximum levels within 30 min. However, HSP70 promoted the upregulation of Beclin-1 expression via Jun N-terminal kinase (JNK) activation in tumor cells and the release of HMGB1 from tumor cells. Inhibition of Beclin-1/c-JNK production prevented the second, but not the first, phase of NF-κB phosphorylation, implicating Beclin-1/c-JNK in the second phase of phosphorylation. HSP70 induced Beclin-1-derived HMGB1 production at 4 h, which occurred before the rise in the second phosphorylation that occurred at 6 h. Exogenous HMGB1 also induced the rapid phosphorylation of NF-κB and upregulated the expression of MMP-9, inhibited the rapid phosphorylation of NF-κB and reduced MMP-9 by receptor for advanced glycation end products (RAGE) inhibitor that prevented HMGB1-induced cell invasion in vitro, which demonstrated that the biological significance of HMGB1/RAGE is key to the second, but not the first, phase of NF-κB phosphorylation in tumor cells. HSP70 triggered a positive feedback loop of NF-κB activation in H22 cells. The second phase of NF-κB phosphorylation mediated by HSP70 is implicated in the increase of tumor cell malignant invasion.

Increased pulse pressure (PP) is an important independent predictor of cardiovascular outcome and acute kidney injury (AKI) after cardiac surgery. The objective of this study was to determine whether elevated baseline PP is associated with postoperative AKI and 30-day mortality after noncardiac surgery.We evaluated 9125 adult patients who underwent noncardiac surgery at Duke University Medical Center between January 2006 and December 2009. Baseline arterial blood pressure was defined as the mean of the first 5 measurements recorded by the automated record keeping system before inducing anesthesia. Multivariable logistic regression analysis was performed to determine whether baseline PP adjusted for other perioperative risk factors was independently associated with postoperative AKI and 30-day mortality.Of the 9125 patients, the baseline PP was40 mm Hg in 1426 (15.6%), 40-80 mm Hg in 6926 (75.9%), and80 mm Hg in 773 (8.5%) patients. The incidence of AKI was 19.8%, which included 8.4% (151 patients) and 4.2% (76 patients) who experienced stage II and III AKI, respectively. In the risk-adjusted model for postoperative AKI, elevated baseline PP was associated with higher odds for postoperative AKI (adjusted odds ratio [OR] for every 20 mm Hg increase in PP, 1.17; 95% confidence interval [CI], 1.10-1.25; P.0001). Also elevated baseline preoperative PP was significantly associated with mild (stage I; OR, 1.19; 95% CI, 1.11-1.27; P.0001), but not with more advanced stages of postoperative AKI or with an incremental risk for 30-day mortality.We found a significant association between elevated baseline PP and postoperative AKI in patients who underwent noncardiac surgery. However, elevated PP was not significantly associated with more advanced stages of postoperative AKI or 30-day mortality in these patients.

Patient-physician communication is an integral part of high-quality patient care and an expectation of the Clinical Learning Environment Review program.Background This quality improvement initiative evaluated the impact of an educational audit and feedback intervention on the frequency of use of 2 tools—business cards and white boards—to improve provider identification.Objective This before-after study utilized patient surveys to determine the ability of those patients to name and recognize their physicians. The before phase began in July 2013. From September 2013 to May 2014, physicians received education on business card and white board use.Methods We surveyed 378 patients. Our intervention improved white board utilization (72.2% postintervention versus 54.5% preintervention, P < .01) and slightly improved business card use (44.4% versus 33.7%, P = .07), but did not improve physician recognition. Only 20.3% (14 of 69) of patients could name their physician without use of the business card or white board. Data from all study phases showed the use of both tools improved patients' ability to name physicians (OR = 1.72 and OR = 2.12, respectively; OR = 3.68 for both; P < .05 for all), but had no effect on photograph recognition.Results Our educational intervention improved white board use, but did not result in improved patient ability to recognize physicians. Pooled data of business cards and white boards, alone or combined, improved name recognition, suggesting better use of these tools may increase identification. Future initiatives should target other barriers to usage of these types of tools.Conclusions

Previous epidemiological studies have established insomnia as a major risk factor for mood, anxiety, and substance use disorders. However, the associations between insomnia with sedative-hypnotic prescriptions and various psychiatric disorders have not been thoroughly examined. The current study involved evaluating the risk of psychiatric disorders, namely schizophrenia, mood, anxiety, somatoform, and substance-related disorders, over a 6-y follow-up period in three groups: patients with insomnia and sedative-hypnotic prescriptions (Inso-Hyp), those with insomnia and without sedative-hypnotic prescriptions (Inso-NonHyp), and those with neither insomnia nor sedative-hypnotic prescriptions (NonInso-NonHyp).We used a historical cohort study design to compare the risk of psychiatric disorders among the three groups. Data regarding these patients were derived from reimbursement claims recorded in Taiwan's National Health Insurance Research Database. Cox proportional hazards regression was used to compare the 6-y risk of subsequent psychiatric disorders among the Inso-Hyp, Inso-NonHyp, and NonInso-NonHyp groups.Compared with the Inso-NonHyp and NonInso-NonHyp group patients, the Inso-Hyp group patients exhibited a higher risk of psychiatric disorders, particularly bipolar disorders (adjusted hazard ratio [AHR]: 7.60; 95% confidence interval [CI]: 5.31-10.89 and AHR: 14.69; 95% CI: 11.11-19.43, respectively). Moreover, among the Inso-Hyp patient group, insomnia prescribed with benzodiazepine, a longer duration of sedative-hypnotic action, and higher doses of sedativehypnotics were significantly associated with a higher risk of depressive and anxiety disorders.The Inso-Hyp group exhibited a higher risk of developing psychiatric disorders than did the Inso-NonHyp and NonInso-NonHyp groups. The results regarding patients with insomnia and sedative-hypnotic prescriptions associated with the risk of psychiatric disorders can serve as a reference for care providers when managing sleep disturbances.

When an innovative (brand-name) drug is going off patent protection, pharmaceutical or generic companies may file an abbreviated new drug application (ANDA) for generic approval. As indicated by the United States Food and Drug Administration (FDA), an approved generic drug can be used as a substitute for the brand-name drug. FDA, however, does not indicate that approved generic drugs of the same brand-name drug can be used interchangeably. As more generic drugs become available in the marketplace, it is a concern whether the approved generic drugs are safe and can be used interchangeably. In this article, we propose two safety margins as new bioequivalence limits for monitoring of drug interchangeability based on a meta-analysis of data obtained from regulatory submissions which have been approved by FDA. In addition to the monitoring of drug interchangeability of generic drugs, the proposed margins can also be extended to address drug interchangeability of biosimilars.

1,25-dihydroxyvitamin D 3 (1,25(OH) 2 D 3 ) is a potent inhibitor of keratinocyte proliferation, as well as a stimulator of epidermal terminal differentiation. In the present studies, we investigated the influence of epidernial growth factor (EGE) and insulin on the antiproliferative and differentiation activities of 1,25(OH) 2 D 3 . Our results indicate the following: (1) EGE caused a dramatic potentiation of the 1,25(OH) 2 D 3 -induced inhibition of 3 H-thymidine incorporation into keratinocytes in a dose-dependent manner; (2) insulin acted antagonistically on the EGF-dependent potentiation of the 1,25(OH) 2 D 3 -induced antiproliferative activity; (3) transforming growth factor-α potentiated 1,25(OH) 2 D 3 -induced antiproliferative activity similar to EGF; (4) the EGF effect was not dependent upon 1,25(OH) 2 D 3 receptor mRNA up-regulation; and (5) removal of insulin from medium supplemented with growth factors significantly potentiated the 1,25(OH) 2 D 3 -induced inhibition on the number of basal cells and the 1,25(OH) 2 D 3 -dependent cornified envelope formation. In conclusion, the antiproliferative activity of 1,25(OH) 2 D 3 in cultured normal human keratinocytes is greatly enhanced by EGF or transforming growth factor-α and reduced by insulin. Insulin also inhibits 1,25(OH) 2 D 3 -induced terminal differentiation of cultured normal human keratinocytes.