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Purpose: Current tumor–node–metastasis (TNM) staging system cannot provide adequate information for prediction of prognosis and chemotherapeutic benefits. We constructed a classifier to predict prognosis and identify a subset of patients who can benefit from adjuvant chemotherapy.Experimental Design: We detected expression of 15 immunohistochemistry (IHC) features in tumors from 251 gastric cancer (GC) patients and evaluated the association of their expression level with overall survival (OS) and disease-free survival (DFS). Then, integrating multiple clinicopathologic features and IHC features, we used support vector machine (SVM)–based methods to develop a prognostic classifier (GC-SVM classifier) with features. Further validation of the GC-SVM classifier was performed in two validation cohorts of 535 patients.Results: The GC-SVM classifier integrated patient sex, carcinoembryonic antigen, lymph node metastasis, and the protein expression level of eight features, including CD3invasive margin (IM), CD3center of tumor (CT), CD8IM, CD45ROCT, CD57IM, CD66bIM, CD68CT, and CD34. Significant differences were found between the high- and low-GC-SVM patients in 5-year OS and DFS in training and validation cohorts. Multivariate analysis revealed that the GC-SVM classifier was an independent prognostic factor. The classifier had higher predictive accuracy for OS and DFS than TNM stage and can complement the prognostic value of the TNM staging system. Further analysis revealed that stage II and III GC patients with high-GC-SVM were likely to benefit from adjuvant chemotherapy.Conclusions: The newly developed GC-SVM classifier was a powerful predictor of OS and DFS. Moreover, the GC-SVM classifier could predict which patients with stage II and III GC benefit from adjuvant chemotherapy. Clin Cancer Res; 24(22); 5574–84. ©2018 AACR.

Figure S6. Kaplan-Meier survival analysis of OS for patients in the external validation cohort according to the GC-SVM classifier stratified by clinicopathological risk factors. P-values were calculated by log-rank test.

Supplemantary figure legends

Table S1. Treatment for patients with gastric cancer in the training, internal and external validation cohorts.

Supplementary materials

Summary: Background: The tumour stroma microenvironment plays an important part in disease progression and its composition can influence treatment response and outcomes. Histological evaluation of tumour stroma is limited by access to tissue, spatial heterogeneity, and temporal evolution. We aimed to develop a radiological signature for non-invasive assessment of tumour stroma and treatment outcomes. Methods: In this multicentre, retrospective study, we analysed CT images and outcome data of 2209 patients with resected gastric cancer from five independent cohorts recruited from two centres (Nanfang Hospital of Southern Medical University [Guangzhou, China] and Sun Yat-sen University Cancer Center [Guangzhou, China]). Patients with histologically confirmed gastric cancer, at least 15 lymph nodes harvested, preoperative abdominal CT available, and complete clinicopathological and follow-up data were eligible for inclusion. Tumour tissue was collected for patients in the training cohort (321 patients), internal validation cohort one (246 patients), and external validation cohort one (128 patients). Four stroma classes were defined according to the protein expression of α-smooth muscle actin and periostin assessed by immunohistochemistry. The primary objective was to predict the histologically based stroma classes by using preoperative CT images. We trained a deep convolutional neural network model using the training cohort and tested the model in the internal and external validation cohort one. We evaluated the model's association with prognosis in the training cohort, two internal, and two external validation cohorts and compared outcomes of patients who received or did not receive adjuvant chemotherapy. Findings: The deep-learning model achieved a high diagnostic accuracy for assessing tumour stroma in both internal validation cohort one (area under the receiver operating characteristic curve [AUC] 0·96–0·98]) and external validation cohort one (AUC 0·89–0·94). The stromal imaging signature was significantly associated with disease-free survival and overall survival in all cohorts (p

Chronic hepatitis B virus (HBV) infection is characterized by the presence of defective viral envelope proteins (hepatitis B surface antigen [HBsAg]) and the duration of infection-most patients acquire the infection at birth or during the first years of life. We investigated the effects of these factors on patients' lymphocyte and HBV-specific T-cell populations.We collected blood samples and clinical data from 243 patients with HBV infection (3-75 years old) in the United Kingdom and China. We measured levels of HBV DNA, HBsAg, hepatitis B e antigen, and alanine aminotransferase; analyzed HBV genotypes; and isolated peripheral blood mononuclear cells (PBMCs). In PBMCs from 48 patients with varying levels of serum HBsAg, we measured 40 markers on nature killer and T cells by mass cytometry. PBMCs from 189 patients with chronic infection and 38 patients with resolved infections were incubated with HBV peptide libraries, and HBV-specific T cells were identified by interferon gamma enzyme-linked immune absorbent spot (ELISpot) assays or flow cytometry. We used multivariate linear regression and performed variable selection using the Akaike information criterion to identify covariates associated with HBV-specific responses of T cells.Although T- and natural killer cell phenotypes and functions did not change with level of serum HBsAg, numbers of HBs-specific T cells correlated with serum levels of HBsAg (r = 0.3367; P.00001). After we performed the variable selection, the multivariate linear regression model identified patient age as the only factor significantly associated with numbers of HBs-specific T cells (P = .000115). In patients younger than 30 years, HBs-specific T cells constituted 28.26% of the total HBV-specific T cells; this value decreased to 7.14% in patients older than 30 years.In an analysis of immune cells from patients with chronic HBV infection, we found that the duration of HBsAg exposure, rather than the quantity of HBsAg, was associated with the level of anti-HBV immune response. Although the presence of HBs-specific T cells might not be required for the clearance of HBV infection in all patients, strategies to restore anti-HBV immune responses should be considered in patients younger than 30 years.

Background The tumor immune microenvironment can provide prognostic and predictive information. A previously validated ImmunoScore of Gastric Cancer (ISGC) evaluates both lymphoid and myeloid cells in the tumor core and invasive margin with immunohistochemical staining of surgical specimens. We aimed to develop a noninvasive radiomics-based predictor of ISGC. Patients and methods In this retrospective study including four independent cohorts of 1778 patients, we extracted 584 quantitative features from the intratumoral and peritumoral regions on contrast-enhanced computed tomography images. A radiomic signature [radiomics ImmunoScore (RIS)] was constructed to predict ISGC using regularized logistic regression. We further evaluated its association with prognosis and chemotherapy response. Results A 13-feature radiomic signature for ISGC was developed and validated in three independent cohorts (area under the curve = 0.786, 0.745, and 0.766). The RIS signature was significantly associated with both disease-free and overall survival in the training and all validation cohorts [hazard ratio (HR) range: 0.296–0.487, all P Conclusion The RIS is a reliable tool for evaluation of immunoscore and retains the prognostic significance in gastric cancer. Future prospective studies are required to confirm its potential to predict treatment response and select patients who will benefit from chemotherapy.

Background Acute-on-chronic liver failure (ACLF) is a clinic syndrome with substantial high short-term mortality. It is very important to stratify patients according to prognosis to decide management strategy. This study aimed to formulate and validate a nomogram model based on blood lipoprotein for prediction of 3-month mortality in patients with hepatitis B virus (HBV)-related ACLF. Methods Data on 393 consecutive patients who were diagnosed as HBV-related ACLF at the Third Affiliated Hospital of Sun Yat-sen University between June 1, 2013, and February 1, 2015, were prospectively collected. Of these, 260 patients who were collected in an earlier period formed the training cohort for the development of nomogram, while 133 patients who were collected thereafter formed the validation cohort for confirming the performance of nomogram. Results Multivariate analysis showed that low density lipoprotein cholesterol (LDL-C), age, prothrombin time, and creatinine were independently associated with 3-month mortality of patients with HBV-related ACLF. Kaplan–Meier survival analysis revealed that the high LDL-C (LDL-C ≥ 1.0 mmol/L, cut-off value) was significantly associated with elevated overall survival (P Conclusion This study highlighted that reduction of serum LDL-C level was an independent risk factor for the survival in patients with HBV-related ACLF, and the nomogram based on serum LDL-C was an accurate and practical model for predicting the 3-month mortality in patients with this disease.

Background: Population-based data on the prognosis of brain metastases at initial diagnosis of ovary cancer (OCBM) are currently lacking. Besides, the effective treatment for OCBM patients is still controversial now. The study aimed to explore the prognostic factors among OCBM. Methods: We retrospectively reviewed the OCBM patients from the Surveillance, Epidemiology, and End Result (SEER) database of the National Cancer Institute to investigate predictors of the presence of OCBM and its' prognostic factors related to all-cause mortality. We employed multivariable logistic and Cox regression analysis. Furthermore, to minimize the impact of potential confounding factors, we conducted a 1:1 propensity score matching (PSM) analysis. Results: A total of 29,512 cases of OC patients entered into the study, including 89 patients with brain metastases of ovarian cancer, which accounted for 0.30% of the entire cohort and 12.02% of the metastatic disease subset. We identified eight factors, including laterality, histology, surgery, radiotherapy, chemotherapy, and extracranial metastatic sites to bone, liver, and lung, as predictors of OCBM based on multivariable logistic regression among the entire cohort. The median survival time of OCBM was 2.0 months, and the interquartile range was 2.0-10.0 mo. The patients who received comprehensive treatment had better prognosis. Based on the multivariable Cox model, marital status, surgery, chemotherapy, and extensive therapy (including RSC, SC, and RC) were identified as predictors of OS. Besides, a new factor (brain metastasis) was identified by 1:1 PSM -based multiple Cox regression, apart from the above prognostic factors for OS. Conclusions: This study provided a population-based estimate of the proportion and prognosis for newly diagnosed ovary cancer with brain metastases. These findings may add materials to guidelines for preliminary screening and optimal treatment of OCBM patients.

INTRODUCTION: Treatments for young patients with gastric cancer (GC) remain poorly defined, and their effects on survival are uncertain. We aimed to investigate the receipt of chemotherapy by age category (18–49, 50–64, and 65–85 years) and explore whether age differences in chemotherapy matched survival gains in patients with GC. METHODS: Patients who were histologically diagnosed with GC were included from a Chinese multi-institutional database and the Surveillance, Epidemiology, and End Results database. There were 5,122 and 31,363 patients aged 18–85 years treated between 2000 and 2014, respectively. Overall survival and stage-specific likelihood of receiving chemotherapy were evaluated. RESULTS: Of the 5,122 and 31,363 patients in China and Surveillance, Epidemiology, and End Result data sets, 3,489 (68.1%) and 18,115 (57.8%) were men, respectively. Younger (18–49 years) and middle-aged (50–64 years) patients were more likely to receive chemotherapy compared with older patients (65–85 years) (64.9%, 56.7%, and 45.4% in the 3 groups from the China data set). Among patients treated with surgery alone, a significantly better prognosis was found in younger and middle-aged patients than their older counterparts; however, no significant differences were found in overall survival among age subgroups in patients who received both surgery and chemotherapy, especially in the China data set. The survival benefit from chemotherapy was superior among older patients (all P < 0.0001) compared with that among younger and middle-aged patients in stage II and III disease. DISCUSSION: Potential overuse of chemotherapy was found in younger and middle-aged patients with GC, but the addition of chemotherapy did not bring about matched survival improvement, especially in the China data set.

Background: Growing evidence indicates that activated hepatic stellate cells (aHSCs) play unexpected roles in regulating immune cells’ function during liver fibrosis. Macrophages feature with inducible plasticity according to the circumstances while patrol for potential pathogens. However, studies seldom investigate whether and how the aHSCs regulate the phenotype and function of macrophages during liver fibrosis. Methods: 96 patients with different stages of liver fibrosis were involved in our study. Metavir score system was used to evaluate the degree of fibrosis. The expression of hepatic CCL2 and CD163 were detected by immunohistochemistry, and the relationship among CD163, CCL2, and fibrosis scores were explored. We co-cultured the aHSCs and THP-1-derived M0-type macrophages (M0MФ) to observe whether aHSCs modulate the expression of CD163 on macrophages in vitro. Furthermore, we treated the M0MФ with aHSCs’ supernatant and investigated the production of CCL2 in aHSCs by ELISA and immunofluorescence assay. To explore whether CCL2/CCR2 axis plays a crucial role in macrophage phenotypic changes during liver fibrosis, we used recombinant CCL2 and its specific receptor antagonist. Moreover, we employed LX2 system to confirm our results. Results: We found that hepatic M2 macrophages (CD163+) infiltration increased in different stages of liver fibrosis (F0-1: 34.95±18.12; F2-3: 77.57±32.48; F4: 99.62±40.84, F0-1 vs. F2-3, P

Additional file 1: Table S1. Univariateanalysis of the primary cohort.

We aimed to evaluate whether radiomic feature-based fluorine 18 (18F) fluorodeoxyglucose (FDG) positron emission tomography (PET) imaging signatures allow prediction of gastric cancer (GC) survival and chemotherapy benefits. Methods: A total of 214 GC patients (training (n = 132) or validation (n = 82) cohort) were subjected to radiomic feature extraction (80 features). Radiomic features of patients in the training cohort were subjected to a LASSO cox analysis to predict disease-free survival (DFS) and overall survival (OS) and were validated in the validation cohort. A radiomics nomogram with the radiomic signature incorporated was constructed to demonstrate the incremental value of the radiomic signature to the TNM staging system for individualized survival estimation, which was then assessed with respect to calibration, discrimination, and clinical usefulness. The performance was assessed with concordance index (C-index) and integrated Brier scores. Results: Significant differences were found between the high- and low-radiomic score (Rad-score) patients in 5-year DFS and OS in training and validation cohorts. Multivariate analysis revealed that the Rad-score was an independent prognostic factor. Incorporating the Rad-score into the radiomics-based nomogram resulted in better performance (C-index: DFS, 0.800; OS, 0.786; in the training cohort) than TNM staging system and clinicopathologic nomogram. Further analysis revealed that patients with higher Rad-scores were prone to benefit from chemotherapy. Conclusion: The newly developed radiomic signature was a powerful predictor of OS and DFS. Moreover, the radiomic signature could predict which patients could benefit from chemotherapy.

To determine whether p21-activated Kinase (PAK) 6 is a prognostic and predictive marker in gastric cancer (GC) and to construct a classifier that can identify a subset of patients who are highly sensitive to 5-fluorouracil/oxaliplatin chemotherapy. We retrospectively analyzed the expression levels of PAK6, cyclooxygenase 2, p21WAF1, Ki-67, excision repair cross-complementing gene 1, and thymidylate synthase in 242 paraffin-embedded GC specimens of the training cohort by immunohistochemistry. Then, we used support vector machine (SVM)-based methods to develop a predictive classifier for chemotherapy (chemotherapy score - CS-SVM classifier). Further validation was performed in an independent cohort of 279 patients. High PAK6 expression was associated with poor prognosis and increased chemoresistance to 5-FU/oxaliplatin chemotherapy. The CS-SVM classifier distinguished patients with stage II and III GC into low- and high-CS-SVM groups, with significant differences in the 5-year disease-free survival (DFS) and overall survival (OS) in chemotherapy patients. Moreover, chemotherapy significantly prolonged the DFS and OS of the high CS-SVM patients in the training and validation cohorts. In conclusion, PAK6 was an independent prognostic factor and increased chemoresistance. The CS-SVM classifier distinguished a subgroup of stage II and III patients who would highly benefit from chemotherapy, thus facilitating patient counseling and individualizing the management.

Current gastric cancer staging alone cannot predict prognosis and adjuvant chemotherapy benefits in stage II and III gastric cancer. Tumor immune microenvironment biomarkers and tumor-cell chemosensitivity might add predictive value to staging. This study aimed to construct a predictive signature integrating tumor immune microenvironment and chemosensitivity-related features to improve the prediction of survival and adjuvant chemotherapy benefits in patients with stage II to III gastric cancer. We used IHC to assess 26 features related to tumor, stroma, and chemosensitivity in tumors from 223 patients and evaluated the association of the features with disease-free survival (DFS) and overall survival (OS). Support vector machine (SVM)–based methods were used to develop the predictive signature, which we call the SVM signature. Validation of the signature was performed in two independent cohorts of 445 patients. The diagnostic signature integrated seven features: CD3+ cells at the invasive margin (CD3 IM), CD8+ cells at the IM (CD8 IM), CD45RO+ cells in the center of tumors (CD45RO CT), CD66b+ cells at the IM (CD66b IM), CD34+ cells, periostin, and cyclooxygenase-2. Patients fell into low- and high-SVM groups with significant differences in 5-year DFS and OS in the training and validation cohorts (all P < 0.001). The signature was an independent prognosis indicator in multivariate analysis in each cohort. The signature had better prognostic value than various clinicopathologic risk factors and single features. High-SVM patients exhibited a favorable response to adjuvant chemotherapy. Thus, this SVM signature predicted survival and has the potential for identifying patients with stage II and III gastric cancer who could benefit from adjuvant chemotherapy.

The aim of the present study was to analyze the efficacy and safety of tenofovir (TDF) treatment for up to 5 years in nucleos(t)ide-naïve chronic hepatitis B (CHB) patients, particularly those with a high viral load, a in real-life scenario. A total of 144 nucleos(t)ide-naïve CHB patients who received TDF monotherapy for at least 3 months were retrospectively analyzed. The primary endpoint measure was the achievement of virological response (VR; undetectable serum HBV DNA, 24 weeks of further TDF therapy to achieve VR. No serious adverse events were reported. In conclusion, long-term TDF treatment of nucleos(t)ide-naïve chronic hepatitis B patients, regardless of high viral load at baseline, was effective and safe in a real-life scenario. Adjustment of TDF monotherapy may be unnecessary in nucleos(t)ide-naïve patients with PVR at 24 weeks.

Treatment options for young patients with gastric cancer (GC) remain poorly defined and their effects on prognosis are unclear. To investigate receipt of chemotherapy by age category (18-49, 50-64, and 65-85 years) and assess whether age differences in chemotherapy matched survival gains among patients with GC. Data were obtained from a Chinese multi-institutional database and the US SEER database. There were 5122 and 31363 patients aged 18 to 85 years treated between 2000 and 2014, respectively. The primary outcome measure was overall survival of patients who only received surgery and those who received both surgery and chemotherapy. The secondary outcome measure was stage-specific likelihood of receiving postoperative chemotherapy. Of the 5122 and 31363 patients in China and SEER datasets, 3489 (68.1%) and 18115 (57.8%) were men, respectively. Younger (18-49 years) and middle-aged (50-64 years) patients were more likely to receive chemotherapy compared with older patients (65-85 years) across all tumor stages. Among patients treated with surgery alone, a significantly better survival was observed for younger and middle-aged patients than their older counterparts, however, no significant differences were observed in survival between age groups among patients who received surgery and chemotherapy, especially in China dataset. The survival benefit from chemotherapy was superior among older patients (all P < 0.0001 for interaction) than among younger and middle-aged patients in stage II and III disease. We found potential overuse of chemotherapy among younger and middle-aged adults, but the addition of chemotherapy did not result in matched survival improvement. Funding: This work was supported by grants from: National Natural Science Foundation of China 81672446, 81600510, 81370575, 81570593. The National Key Research and Development Program of China 2017YFC0108300. Natural Science Foundation of Guangdong Province, 2014A030313131. Science and Technology Planning Project of Guangzhou, 2014B020228003, 2014B030301041, 2015A030312013. Key Clinical Specialty Discipline Construction Program. Director’s Foundation of Nanfang Hospital, 2016B010. Declaration of Interest: None of the authors had any potential conflicts of interest, including relevant financial interests, activities, relationships, and affiliations. Ethical Approval: The institutional review boards at Nanfang Hospital of Southern Medical University, First Affiliated Hospital of Sun Yat-sen University, and Sun Yat-sen University Cancer Center approved the retrospective analysis of anonymous data and waived the need to obtain patient informed consent.

Purpose: Current tumor–node–metastasis (TNM) staging system cannot provide adequate information for prediction of prognosis and chemotherapeutic benefits. We constructed a classifier to predict prognosis and identify a subset of patients who can benefit from adjuvant chemotherapy. Experimental Design: We detected expression of 15 immunohistochemistry (IHC) features in tumors from 251 gastric cancer (GC) patients and evaluated the association of their expression level with overall survival (OS) and disease-free survival (DFS). Then, integrating multiple clinicopathologic features and IHC features, we used support vector machine (SVM)–based methods to develop a prognostic classifier (GC-SVM classifier) with features. Further validation of the GC-SVM classifier was performed in two validation cohorts of 535 patients. Results: The GC-SVM classifier integrated patient sex, carcinoembryonic antigen, lymph node metastasis, and the protein expression level of eight features, including CD3invasive margin (IM), CD3center of tumor (CT), CD8IM, CD45ROCT, CD57IM, CD66bIM, CD68CT, and CD34. Significant differences were found between the high- and low-GC-SVM patients in 5-year OS and DFS in training and validation cohorts. Multivariate analysis revealed that the GC-SVM classifier was an independent prognostic factor. The classifier had higher predictive accuracy for OS and DFS than TNM stage and can complement the prognostic value of the TNM staging system. Further analysis revealed that stage II and III GC patients with high-GC-SVM were likely to benefit from adjuvant chemotherapy. Conclusions: The newly developed GC-SVM classifier was a powerful predictor of OS and DFS. Moreover, the GC-SVM classifier could predict which patients with stage II and III GC benefit from adjuvant chemotherapy. Clin Cancer Res; 24(22); 5574–84. ©2018 AACR.

Background: The risk of lymph node positivity (LN ) in gastric cancer (GC) impacts therapeutic recommendations. The aim of this study was to quantify the effect of younger age on LN . Methods: Data from a Chinese multi-institutional database and the US SEER database on stage I to III resected GC were analyzed for the relationship between age and LN status. The association of age and LN status was examined with logistic regression separately for each T stage, adjusting for multiple covariates. Poisson regression was used to evaluate age and number of LN . Findings: 4,905 and 14,877 patients were identified in the China and SEER datasets respectively. 479 (9.8%) patients were under age 40 years, with 768 (15.7%) between age 40 and 49 years in China dataset, and 416 (2.8%) patients were under age 40 years, with 1176 (7.9%) between age 40 and 49 years in SEER dataset. Both datasets exhibited significantly proportional decreases of N3a and N3b LN with age increasing. Patients younger than age 40 years were more likely to show LN compared with the reference age 60 to 69 years. The youngest patients had the highest ORs of N1, N2, N3a, and N3b vs N0 LN within T4 stage of China dataset and T3 stage of SEER dataset, the values of ORs decreased with increasing age. Young age was a predictor of an increased number of LNs positive for each T stage. Interpretation: In the two large datasets, young age at diagnosis is associated with an increased risk of LN . Funding: This work was supported by grants from: National Natural Science Foundation of China 81672446, 81370575, 81570593. Natural Science Foundation of Guangdong Province, 2014A030313131. Science and Technology Planning Project of Guangzhou, 2014B020228003, 2014B030301041, 2015A030312013. Science and Technology Program of Guangzhou, 158100076, 201400000001-3. Public welfare in Health Industry, National Health and Family Planning Commission of China (201402015, 201502039). Key Clinical Specialty Discipline Construction Program. Declaration of Interest: The authors declare that they have no conflict of interest. Ethical Approval: This study was approved by the research ethics committees at the 3 participating centers, and the informed consent requirement was waved.

Background The optimal treatment for locally advanced borderline-resectable squamous cell carcinoma (SCC) of the oesophagus is unclear. This phase II trial assessed the safety and efficacy of paclitaxel in combination with cisplatin and 5-fluorouracil (TPF) induction chemotherapy and subsequent surgery for locally advanced borderline-resectable esophageal SCC (NCT02976909). Methods Patients with clinical T4 and/or bulky lymphadenopathy that may invade nearby organs were eligible. Treatment started with 2 to 4 cycles of TPF induction chemotherapy (paclitaxel 135mg/m2 on day 1, cisplatin 75mg/m2 on Day 1; 5-fluorouracil 4g/m2 for 120 hours continuous infusion from Day 1 to Day 5), followed by surgery if resectable, or by radical concurrent chemoradiotherapy if unresectable. The primary endpoint was pathologically proven complete resection (R0) rate. Results From November 2016 to December 2018, 47 patients were enrolled. After 2 to 4 cycles TPF chemotherapy, surgery was performed in 27 patients (57.4%), 10 (21.3%) patients received radical concurrent chemoradiotherapy. R0 resection was confirmed in 25 patients (53.2%). Pathologic complete response after chemotherapy was confirmed in 4 patients (8.5%). During chemotherapy, the most common grade 3 or 4 toxicities were neutropenia (29.8%), leucopenia (21.3%), febrile neutropenia (4.3%) and stomatitis (4.3%). No serious postoperative complications were observed in patients undergoing surgery. Conclusions TPF induction chemotherapy followed by surgery as a multidisciplinary treatment strategy showed promising signs of tolerability and short-term efficacy in patients with locally advanced borderline-resectable esophageal SCC. Long-term survival is still being followed up. Legal entity responsible for the study Yuhong Li. Funding Guangdong Esophageal Cancer institute. Disclosure All authors have declared no conflicts of interest.