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Background Circumnutation (Darwin et al., Sci Rep 10(1):1–13, 2000) is the side-to-side movement common among growing plant appendages but the purpose of circumnutation is not always clear. Accurately tracking and quantifying circumnutation can help researchers to better study its underlying purpose. Results In this paper, a deep learning-based model is proposed to track the circumnutating flowering apices in the plant Arabidopsis thaliana from time-lapse videos. By utilizing U-Net to segment the apex, and combining it with the model update mechanism, pre- and post- processing steps, the proposed model significantly improves the tracking time and accuracy over other baseline tracking methods. Additionally, we evaluate the computational complexity of the proposed model and further develop a method to accelerate the inference speed of the model. The fast algorithm can track the apices in real-time on a computer without a dedicated GPU. Conclusion We demonstrate that the accuracy of tracking the flowering apices in the plant Arabidopsis thaliana can be improved with our proposed deep learning-based model in terms of both the racking success rate and the tracking error. We also show that the improvement in the tracking accuracy is statistically significant. The time-lapse video dataset of Arabidopsis is also provided which can be used for future studies on Arabidopsis in various takes.

Bladder cancer-associated transcript 1 (BLACAT1) is a novel identified long noncoding RNA (lncRNA) in bladder cancer, and has been suggested to function as an oncogenic lncRNA in several types of human cancer. However, its involvement in the progression of small-cell lung cancer (SCLC) remained unknown. The aim of our study was to investigate the clinical value and biological function in SCLC. In our results, BLACAT1 expression was increased in SCLC tissues and cell lines compared with paired adjacent normal tissues and bronchial epithelial cell lines, respectively. In addition, BLACAT1 high-expression was obviously associated with advanced clinical stage, large tumor size, more lymph node metastasis, present distant metastasis, and poor prognosis. Furthermore, multivariate analysis indicated that high-expression of BLACAT1 acted as an independent poor prognostic factor for overall survival in SCLC cases. The loss-of-function studies suggested that of BLACAT1 suppressed SCLC cell proliferation, migration, and invasion, and induced G0/G1 phase arrest. In conclusion, BLACAT1 is associated with the malignant status and prognosis in patients with SCLC, and functions as an oncogenic lncRNA in regulating cell proliferation and motility, suggesting BLACAT1 may act as a potential target for SCLC prevention and treatment.

// Yixiang Mao 1, 2, 5 , Jianjun Shen 4 , Yue Lu 4 , Kevin Lin 4 , Huamin Wang 3 , Yanan Li 2 , Ping Chang 2 , Mary G. Walker 4 and Donghui Li 2 1 Department of Oncology, The First Affiliated Hospital of Soochow University, Suzhou 215007, China 2 Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA 3 Department of Pathology and Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA 4 Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, Texas 78957, USA 5 Department of Medical Oncology, Fudan University Shanghai Cancer Center, Shanghai 200032, China Correspondence to: Donghui Li, email: dli@mdanderson.org Yixiang Mao, email: maoyix@suda.edu.cn Keywords: pancreatic cancer, RNA sequencing, transcriptome, pathway analysis Received: June 03, 2016 Accepted: February 27, 2017 Published: March 22, 2017 ABSTRACT Gene expression microarrays have identified many tumor markers and therapeutic targets for pancreatic ductal adenocarcinoma (PDAC). However, microarray profilings have limited sensitivity and are prone to cross-hybridization between homologous DNA fragments. Here, we perform a transcriptome analysis of paired tumor and adjacent benign pancreatic tissues from 10 patients who underwent resection for PDAC. We identify a total of 2736 differentially expressed genes (DEGs) with false discovery rate less than 0.05, including 1554 upregulated, 1182 downregulated, and 6 microRNAs (miR-614, miR-217, miR-27b, miR-4451, miR-3609, and miR-612). Overexpression of five DEGs, i.e. KRT16 , HOXA10 , CDX1 , SI , and SERPINB5 in tumors is confirmed by RT-PCR in 20 additional tissues. Overexpression of KRT16 in PDAC is also verified on protein level. In addition, top canonical pathways such as granulocyte adhesion and diapedesis pathway have been identified. Our study represents a comprehensive characterization of the PDAC transcriptome and provides insight to the mechanisms of pancreatic carcinogenesis and potential biomarkers and novel therapeutic targets for pancreatic cancer.

Importance Prospective assessment of treatments known to benefit patients in global clinical trials in specific racial groups is essential. Objective To compare the efficacy, safety, and tolerability of adding pertuzumab to trastuzumab and docetaxel vs placebo, trastuzumab, and docetaxel in Asian patients with ERBB2-positive early or locally advanced breast cancer. Design, Setting, and Participants This multicenter, double-blind, placebo-controlled phase 3 trial enrolled 329 women with ERBB2-positive early (T2-3, N0-1, M0) or locally advanced breast cancer (T2-3, N2 or N3, M0; T4, any N, M0) and primary tumor larger than 2 cm from March 14, 2016, to March 13, 2017. Analysis of the primary end point was performed on an intention-to-treat basis. Interventions Before surgery, patients received 4 cycles of intravenous pertuzumab (840-mg loading dose and 420-mg maintenance doses), trastuzumab (8-mg/kg loading dose and 6-mg/kg maintenance doses), and docetaxel (75 mg/m2) or intravenous placebo, trastuzumab, and docetaxel every 3 weeks. After surgery, patients received 3 cycles of intravenous fluorouracil, epirubicin, and cyclophosphamide followed by 13 cycles of the same intravenous anti-ERBB2 therapy (pertuzumab and trastuzumab or placebo and trastuzumab) for up to 1 year. Main Outcomes and Measures The primary end point was independent review committee–assessed total pathologic complete response rate. The 2-sided Cochran-Mantel-Haenszel test, stratified by disease category and hormone receptor status, was used to compare rates between treatment groups. Results In total, 329 female patients were randomized (pertuzumab, 219; and placebo, 110; mean [SD] age, 48.8 [9.5] years). In the intention-to-treat population, total pathologic complete response rates were 39.3% (86 of 219) in the pertuzumab group and 21.8% (24 of 110) in the placebo group (difference, 17.5% [95% CI, 6.9%-28.0%];P = .001). Of the most common grade 3 or higher adverse events, there was a higher incidence of neutropenia in the pertuzumab group (83 of 218 [38.1%] vs 36 of 110 [32.7%]). Serious adverse events were reported in 10.1% of patients (22 of 218) in the pertuzumab group and 8.2% of patients (9 of 110) in the placebo group. Conclusions and Relevance Treatment with pertuzumab, trastuzumab, and docetaxel resulted in a statistically significant improvement in the total pathologic complete response rate vs placebo, trastuzumab, and docetaxel for the neoadjuvant treatment of ERBB2-positive early or locally advanced breast cancer in Asian patients. Safety data were in line with the known pertuzumab safety profile and generally comparable between treatment groups. The PEONY trial adds to the totality of data showing the benefit of the pertuzumab regimen. Trial Registration ClinicalTrials.gov identifier:NCT02586025

In the APHINITY study (NCT01358877, BIG 4-11/BO25126/TOC4939G), pertuzumab added to trastuzumab and chemotherapy significantly improved invasive disease-free survival as adjuvant treatment for patients with HER2-positive early breast cancer. The objective of this analysis was to assess the pharmacokinetics of pertuzumab in combination with trastuzumab in Chinese patients with early breast cancer. Samples for pertuzumab and trastuzumab pharmacokinetic analysis were taken from Chinese patients during cycle 1 of treatment and at steady-state in cycle 10. Noncompartmental analysis was used to estimate minimum and maximum serum concentrations (C-max and C-min), area under the concentration-time curve, clearance, and other pharmacokinetic parameters. In 15 patients, mean steady-state C-max and C-min pertuzumab serum concentrations (368 +/- 177 mu g/ml, and 122 +/- 47 mu g/ml, respectively) were numerically higher than observed previously in a pharmacokinetic analysis of the global population in APHINITY and in patients treated in the metastatic setting. The geometric mean ratio and corresponding 90% confidence interval for trastuzumab C-max and C-min in the presence (n = 15) or absence (n = 17) of pertuzumab were 104.6 (91.09-120) and 98.23 (84.58-114), respectively, indicating no apparent impact of pertuzumab on the pharmacokinetics of trastuzumab. Increases in pertuzumab C-max and C-min were not associated with an increase in adverse events. The APHINITY Chinese pharmacokinetic substudy analysis supports the dosing regimen for pertuzumab (840 mg loading dose followed by 420 mg maintenance doses every 3 weeks administered by intravenous infusion) in a Chinese HER2-positive early breast cancer patient population.

// Chang Liu 1, 4 , Cathy Eng 1 , Jianjun Shen 3 , Yue Lu 3 , Yoko Takata 3 , Amir Mehdizadeh 1 , George J. Chang 2 , Miguel A. Rodriguez-Bigas 2 , Yanan Li 1 , Ping Chang 1 , Yixiang Mao 1 , Manal M. Hassan 1 , Fangyu Wang 4 , Donghui Li 1 1 Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA 2 Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA 3 Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, Texas, USA 4 Department of Gastroenterology and Hepatology, Jinling Hospital, Southern Medical University, Nanjing, China Correspondence to: Donghui Li, email: dli@mdanderson.org Fangyu Wang, email: wangfy65@nju.edu.cn Keywords: exosome, microRNA, colon cancer, qRT-PCR, Illumina RNA sequencing Received: April 22, 2016 Accepted: October 11, 2016 Published: October 24, 2016 ABSTRACT Purpose: The study was aimed to evaluate the prognostic or predictive value of serum exosomal microRNAs (miRNAs) for tumor recurrence and response to adjuvant therapy in stage II and stage III colon cancer. Results: 145 differentially expressed mature miRNAs were identified ( P

// Zuguang Xia 1, * , Xiaoyan Jia 3, * , Kai Chen 2 , Dapeng Li 2 , Jing Xie 2 , Hong Xu 2 , Yixiang Mao 2, 3 1 Department of Medical Oncology, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China 2 Department of Oncology, The First Affiliated Hospital of Soochow University, Suzhou, China 3 Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA * These authors contributed equally to this work Correspondence to: Zuguang Xia, email: zuguang_xia@163.com Hong Xu, email: 13301549066@163.com Yixiang Mao, email: maoyix@gmail.com Keywords: pancreatic adenocarcinoma, postoperative radiation therapy, lymph node metastasis, survival Received: March 10, 2016 Accepted: April 16, 2016 Published: May 26, 2016 ABSTRACT The benefit of combining postoperative radiation therapy (PORT) with chemotherapy for resected patients with pancreatic adenocarcinoma is controversial. We sought to determine the effects of PORT on survival in patients with pancreatic adenocarcinoma who underwent primary site surgery. Patients with pancreatic adenocarcinoma receiving primary tumor surgery between 1988 and 2012 were identified from the Surveillance, Epidemiology and End Results (SEER) database. We estimated the association between PORT and other clinicopathologic factors and survival. In total, 5304 patients were identified who underwent pancreatic resection including 2093 patients who had PORT and 3211 patients who had no PORT. Median overall, cancer-specific, and other-cause survival were 19.0, 20.0, and 196.0 months, respectively, with PORT versus 14.0, 15.0, and 163.0 months, respectively, without PORT (all P < 0.001). Subset analysis revealed that the benefit of PORT was limited to patients with N1 disease. Median overall, cancer-specific, and other-cause survival for patients with N1 disease were 18.0, 18.0, and NA months, respectively, with PORT versus 12.0, 13.0, and 154.0 months, respectively, without PORT (all P < 0.001). Regardless the number of positive lymph node count (PLN) and lymph node ratio (LNR), PORT was always associated with increased survival on multivariate analysis in patients with N1 disease (all P < 0.001). In summary, survival benefits might be obtained from PORT on lymph node positive patients with pancreatic adenocarcinoma.

Additional file 1: Table S1. Post-study anti-cancer therapies used by at least 2 patients (ITT population).

Virtual reality (VR) applications have become popular recently and rapidly commercialized. The behaviors of users watching 360-degree omni-directional videos have not been fully investigated. In this paper, a dataset of view trajectories for users watching 360-degree videos over computer (desktop/laptop) environment is presented. The dataset includes view center trajectory data collected from viewers watching twelve 360-degree videos over the computer, using mouse to navigate and explore the environment. The selected videos cover a variety of contents, leading to different navigation patterns and behaviors. Based on the dataset, we demonstrate that the viewers share similar viewing patterns over certain 360-degree video category. We also compare the view motion patterns and statistics with prior datasets captured using head-mounted display (HMD). The dataset has been made available online, to facilitate the studies on 360-degree video view prediction, content saliency analysis and VR streaming, etc.

Background. Diabetic patients with breast cancer receiving metformin and neoadjuvant chemotherapy have a higher pathologic complete response rate than do diabetic patients not receiving metformin, but findings on salvage treatment have been inconsistent. We performed a meta-analysis to assess the effect of adding metformin to standard therapy on the prognosis of breast cancer patients with diabetes. Methods. We searched PubMed, Embase, Web of Science (Thomson Scientific), China Knowledge Resource Integrated Database, VIP journal integration platform, and Chinese BioMedical Literature Database from inception to January 10, 2015, without language restrictions, including references related to metformin, breast cancer, and prognosis. We performed the meta-analysis using a random-effects model, with hazard ratios (HRs) and 95% confidence intervals (95% CIs) as effect measures. Results. A total of 11 studies consisting of 5,464 breast cancer patients with diabetes were included, comprising 2,760 patients who had received metformin and 2,704 patients who had not. The meta-analysis showed that metformin was associated with better overall survival times (HR: 0.53; 95% CI: 0.39-0.71) and cancer-specific survival times (HR: 0.89; 95% CI: 0.79-1.00). Subgroup analysis revealed that metformin improved the overall survival by 65% after adjusting for hormone receptor expression (HR: 0.35; 95% CI: 0.15–0.84). Taking metformin after the diagnosis of breast cancer was still associated with prolonged overall survival. Conclusion. The use of metformin in standard cancer therapy might improve both overall and cancer-specific survivals of diabetic patients with breast cancer. Implications for Practice: Diabetic patients with breast cancer receiving metformin and neoadjuvant chemotherapy have a higher pathologic complete response rate than diabetic patients not receiving metformin, but findings on salvage treatment have been inconsistent. The meta-analysis showed that metformin was associated with better overall survival times and cancer-specific survival times. Subgroup analysis revealed that metformin improved the overall survival by 65% after adjusting for hormone receptor expression. Taking metformin after the diagnosis of breast cancer was still associated with prolonged overall survival. The findings of this study highlight the potential usage of metformin in diabetic patients with breast cancer.

B7-H1 and B7-H3, two members of the B7 family that are thought to regulate T-cell activation, are expressed in human non-small cell lung cancer (NSCLC). However, their prognostic significance is poorly understood. In the present study we reported that B7-H1 and B7-H3 were expressed in 96/128 (72.7%) and 89/128 (69.5%) samples, respectively. B7-H1 and B7-H3 expression and the number of infiltrating T-cell intracellular antigen-1+ and interferon-γ+ cells in NSCLC tissues were significantly higher than those in the adjacent tissues (p

CD8 CTLs have been accountable for the major effector cells responsible for the rejection of tumor cells. And CD40 signaling and IL-12 have been shown to be the essential pathways involved in the activation process. Immunizing mice with dendritic cells transduced with an adenovirus expressing the human melanoma antigen gp100, an immunization strategy of xenoimmunization, stimulated potent tumor protection dependent on effective CD4 T cells in the absence of CD8 T cells. Further studies revealed that neither CD40 signaling nor IL-12 was indispensable for the activation of dendritic and CD4 T cells in this model. Stimulation of effective antitumor immunity targeting the self-antigen did not elicit autoimmunity. The implications of this study were discussed.

BACKGROUND AND OBJECTIVE B7-H1, a member of B7 family, is expressed in tumor cells and has emerged as an important immune modulator capable of suppressing host immunity by inhibiting T cells function. This study was to probe into the correlation between the expression level of B7-H1 protein in pancreatic carcinoma tissues and clinicopathological characteristics and prognosis. METHODS The expression of B7-H1 was measured in 40 cases of pancreatic carcinoma tissues and 10 cases of normal corresponding paracarcinoma tissues by immunohistochemistry. The relationship between the expression level of B7-H1 and clinicopathological characteristics and survival was analyzed. RESULTS The positive rate of B7-H1 was significantly higher in the tumor tissues [45.00% (18/40)] than in the normal corresponding paracarcinoma tissues [0(0/10)] (P

Concurrent diabetes has been linked with an increased risk of death in many cancers, but findings in pancreatic cancer have been inconsistent. We performed a systematic review and meta-analysis to assess the effect of diabetes on survival in patients with pancreatic cancer. Of 4, 463 original articles, 41 were included in the review; 29 studies with 33 risk estimates were included in the meta-analysis. In the overall comparison of patients with pancreatic cancer and diabetes with their nondiabetic counterparts, the former had significantly higher all-cause mortality (pooled HR: 1.13; 95% CI: 1.04–1.22). Subgroup analyses showed that diabetes was associated with poor survival in patients with resectable disease (HR: 1.37; 95% CI: 1.15–1.63) but not in those with unresectable disease (HR: 1.07; 95% CI: 0.89–1.29). The HR (95% CI) was 1.52 (1.20–1.93) for patients with new-onset diabetes (≤2 years of diabetes duration) and 1.22 (0.83–1.80) for those with longstanding diabetes (>2 years). Diabetes was associated with higher mortality overall in patients with pancreatic cancer. The effect of diabetes on overall survival was associated with the stages of tumor and the duration of diabetes.

B7-H3 and B7-H4 belong to the peripheral membrane protein B7 family and are hypothesized to regulate immunity. These proteins are expressed in human pancreatic cancer (PC), but their prognostic significance is poorly understood. The present study examined the association between B7-H3 and B7-H4 expression and the overall survival time in patients with PC that underwent surgery at the Second Affiliated Hospital to Zhengzhou University between April 2000 and January 2009. Immunohistochemical analysis demonstrated that B7-H3 and B7-H4 were expressed in 35 (88%) and 30 (75%) tumor tissue samples, respectively, which were obtained from 40 patients with PC. Statistical analysis revealed that B7-H3 expression was associated with an early tumor-node-metastasis stage (stage I and II; P

association between long-term diabetes and poor overall survival among patients with diabetes and pancreatic cancer in three longitudinal studies. The authors reported a 40% to 50% increased risk of death for patients with long-term diabetes (duration of 4 years) compared with those without diabetes (P .05) in two prospective cohort studies 2,3 and a clinic-based study. 1 In contrast, no statistically significant association between short-term diabetes (duration of 4 years) and survival was observed. Thus, the authors concluded that there is a link between chronic glucose intolerance and pancreatic cancersurvival,whereasshort-termdiabetesdoesnotportendaworse prognosis among patients with pancreatic adenocarcinoma. We have a concern about the authors’ conclusion regarding the association between short-term diabetes and survival. Because the information on diabetes was collected via a biennial questionnaire in the cohort studies, it is possible that the number of patients with short-term diabetes was greatly underestimated, which may have biased the risk estimate toward null. The prevalence of patients with short-term diabetes was 4.67% (47 of 1,006) and 18.65% (72 of 386), whereas the prevalence of patients with any type of diabetes was 17.99% (181 of 1,006) and 31.35% (121 of 386) in the cohort and the clinic-based studies, respectively. The prevalence of diabetes in the clinic-based study was quite similar to the numbers previously reported by other studies. Short-term diabetes was associated with a nonsignificant but increased risk of death in this clinic series (hazard ratio [HR], 1.22; 95% CI, 0.89 to 1.67). In our recently completed studyof1,328patientswithpathologicallyconfirmedpancreaticductal adenocarcinoma of all stages at MD Anderson Cancer Center, we foundthatshort-termdiabetes(durationof3years)wasassociated with a significantly increased risk of death (HR, 1.23; 95% CI, 1.09 to 1.40).Wehavealsoconductedameta-analysisof29studiesondiabetesandoverallsurvivalofpatientswithpancreaticcancerandfounda reduced survival of patients with diabetes in 25 clinic-based studies (pooled HR, 1.13; 95% CI, 1.02 to 1.25), but not in four cohort studies 4-7 (pooled HR, 1.12; 95% CI, 0.95 to 1.32). The prevalence of patients with diabetes was 28.61% (4,133 of 14,446) and 20.92% (1,124 of 5,372) in the clinic and cohort studies, respectively. These observations suggest that short-term diabetes is associated with a higher risk of death in patients with pancreatic cancer. The inconsistentobservationswithrespecttotheassociationbetweendiabetesand

Cancer metastasis is a highly coordinated and dynamic multistep process in which cancer cells interact with a variety of host cells. Morphological studies have documented the association of circulating tumor cells with host platelets, where a surface coating of platelets protects tumor cells from mechanical trauma and the immune system. Cantharidin is an active constituent of mylabris, a traditional Chinese medicine. Cantharidin and norcantharidin are potent protein phosphatase 2A (PP2A) inhibitors that exhibit in vitro and in vivo antitumor activity against several types of cancer, including breast cancer. We investigated whether cantharidin and norcantharidin could repress the ability of MCF-7 breast cancer cells to adhere to platelets. Using MTT, clone formation, apoptosis, adhesion and wound-healing assays, we found that cantharidin and norcantharidin induced apoptosis and repressed MCF-7 cell growth, adhesion and migration. Moreover, we developed a flow cytometry-based analysis of tumor cell adhesion to platelets. We proved that cantharidin and norcantharidin repressed MCF-7 cell adhesion to platelets through downregulation of α2 integrin, an adhesion molecule present on the surface of cancer cells. The repression of α2 integrin expression was found to be executed through the protein kinase C pathway, the activation of which could have been due to PP2A inhibition.

Cancer metastasis is a highly coordinated and dynamic multistep process in which cancer cells interact with a variety of host cells. Morphological studies have documented the association of circulating tumor cells with host platelets. Tumor cell-induced platelet aggregation (TCIPA) contributes significantly to hematogenous metastasis; however, the molecular mechanisms involved in breast cancer TCIPA are poorly characterized. In this study, MCF-7 metastatic human breast cancer cells induced dose-dependent aggregation of washed platelets. Four major platelet activation pathways, glycoprotein (GP)-Ib-IX, GPIIb/IIIa, thromboxane (TX)-A2 and adenosine diphosphate (ADP) were activated during TCIPA and were inhibited by their respective inhibitors, 7E3, SZ-1, aspirin and apyrase. Pretreatment of platelets with 7E3, SZ-1 or apyrase significantly inhibited TCIPA, while pretreatment with aspirin had no effect. Moreover, combined pretreatment of platelets with 7E3, SZ-1 and apyrase significantly inhibited TCIPA, compared to single inhibitors. Combinations of antiplatelet drugs may represent a promising strategy to prevent cancer metastasis.

The prevalence of CD4+CD25+ regulatory T cells (Tregs) and other T-cell subsets in gastrointestinal cancer patients treated with chemotherapy was investigated.The frequencies of Tregs and other T-cell subsets in the peripheral blood of 25 healthy donors and 104 patients with gastrointestinal cancer were measured by flow cytometry. Their plasma cytokine levels were determined by enzyme-linked immunosorbent assay (ELISA). The percentages of the T-cell subsets and their correlation with the outcomes of the gastrointestinal cancer patients after chemotherapy were evaluated.Not only was the prevalence of Tregs in the peripheral blood of gastrointestinal cancer patients significantly higher than that in healthy donors, but it also increased in parallel with tumor progression. Among patients with advanced disease, 3 weeks after chemotherapy, those with higher percentages of Tregs had a poorer prognosis. Interleukin (IL)-2 production showed a reverse trend to Tregs, whereas IL-10 and tumor necrosis factor (TNF)-alpha did not change significantly.The relative increase in CD4+CD25+ Tregs may be related to immunosuppression and tumor progression in patients with gastrointestinal cancer. Chemotherapy in patients with advanced disease showed that immunosuppression is enhanced early (about 1-2 weeks) after chemotherapy, and increased Tregs 3 weeks after chemotherapy correlated with a poor prognosis.