bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Tourist Satisfaction Index Based on Structural Equation Model
Authors: Peng Yan , Jiang Xiaolin
Published: June 2022

In the modern economy and society, traveling abroad has gradually become one of the ways for people to enjoy life. With the continuous development of tourism and the change of tourists’ consumption concepts, tourists’ satisfaction has been received by more and more tourism managers and operators. It pays attention to providing tourism products and services from the perspective of tourists and meets the personalized and diversified needs of tourists. Tourist satisfaction has become an important indicator to measure the level of tourism in a tourist destination. According to the structural equation model, this paper constructs the tourist satisfaction index analysis index, which can truly and efficiently detect the tourist satisfaction index, which is convenient for the scenic spot to find out the areas that need to be improved, and enhance the new attraction and service level of the scenic spot. The research results of the article show that (1) there is no significant difference in the satisfaction of tourists of different genders. In terms of influencing factor indicators, individual factors have not passed the

T

test. The difference in satisfaction of tourists of different ages is not obvious, but there are also individual factors that do not pass the significance test in terms of influencing factors. For the actual perceived quality, the observation index with a larger contribution rate to principal component 2 is mainly the tourists’ evaluation of the satisfaction of the tourist attractions. (2) The test result of the structural equation-based satisfaction analysis model proposed in the article is the highest among the four models. According to the error curve, it can be concluded that with the increasing complexity of the data, for the other three models, the evaluation error of the tourist satisfaction evaluation model is getting larger and larger, and the errors of the models proposed in the article are basically controlled within 0.4. Therefore, it can be concluded that the satisfaction evaluation model based on structural equations can reduce the error of tourist satisfaction.
Research on the Key Technology of Web Data Extraction and Mining Based on the Probability Distribution
Authors: Yang Jinqiao , Yang Binghui , Sun Qi , Yan Shi , Miao Yuxin
Published: June 2022

In the development of internet technology innovation, the quantity of big data information continues to rise, showing the characteristics of dynamic, heterogeneous, massive, and so on. How to explore valuable and potential knowledge from the network system is the main topic of research and scholars. Based on the understanding of web data extraction and web mining technology, this paper makes a systematic study of web text mining, proposes the most commonly used hidden Markov model in probability distribution, and constructs the corresponding text mining method. The hidden Markov model is a statistical model, which is mainly used to represent a Markov process with unknown parameters. The difficulty in practical application lies in how to define the hidden parameters of the process from observable parameters and then use these parameters to conduct in-depth research. Finally, the practical results show that the hidden Markov model can not only obtain the information of different regions but also deeply analyze the data set, which proves the feasibility of this research technology.
Modeling and Control of Wind Speed in Renewable Energy Power Generation and Wind Power Generation Systems
Authors: Shan Junwei , Lu Shixuan , Liu Shu , Shi Hang
Published: June 2022

Wind energy is one of the most used clean energy sources in renewable energy, and its renewable and sustainable nature is one of the reasons why it is used for power generation. In the current environment where all countries in the world are facing energy problems, research on wind power generation systems is also increasing. This article aims to study the problem of modeling and controlling wind speed in the wind power generation system of renewable energy power generation. To this end, this article proposes a modeling method for wind power generation systems, which can be used to study the momentum problems in wind power generation and the mechanical torque of the generator. And at the end of the article, related experiments and analysis are designed to explore and compare its operating cost, speed, and wind wheel speed. The experimental results in this paper show that through effective modeling and control of its wind speed, the economic risks in the actual wind power generation system can be controlled, with a maximum reduction of 24%, and the actual operating cost is also reduced by 8.66%, so wind power has high practical value.
Antioxidant Activity and Inhibitory Effects of Black Rice Leaf on the Proliferation of Human Carcinoma Cells
Authors: Thepthanee Chorpaka , Liu Chan-Chiung , Yu Hsu-Sheng , Huang Ho-Shin , Yen Chia-Hung , Li Yen-Hsien , Lee Maw-Rong , Liaw Ean-Tun
Published: June 2022

The leaves of black rice, well‐known as postharvest agricultural waste, contain a rich source of antioxidants with multiple benefits for human health. In the present study, the ethyl acetate fraction obtained from black rice leaf was separated into five subfractions using Sephadex LH‐20 column chromatography, and their antioxidant and anticancer activities were investigated. The results revealed that among all the subfractions, subfraction 5 (Sub5) showed the highest total phenolic and flavonoid values. The antioxidant activity was also superior in Sub5 (the IC50 values are 3.23, 31.95, and 72.74 μg/mL, in the DPPH, ABTS, and reducing power assays, respectively) compared to the other subfractions. All subfractions, in a time‐dependent manner, inhibited the proliferation of hepatoma (HepG2), breast (MCF‐7), and colorectal (Caco‐2) cancer cells, especially the Sub5. Thus, Sub5 was employed to conduct the cell cycle and cell apoptosis by flow cytometry. Sub5 significantly increased the accumulation of cells at the Sub‐G1 phase in HepG2 cells (44.5%, at 48 h). Furthermore, it could trigger annexin V‐detected apoptosis through mitochondrial and death receptor pathways accompanied by the suppression of PI3K/Akt and Erk signaling pathways. In addition, HPLC‐DAD‐MS/MS was conducted to characterize the bioactive constituents in the most potent antioxidant, cytotoxic, and apoptosis‐inducing subfraction. Conclusively, Sub5 may have high potential as functional dietary supplements to inhibit the development of HepG2 liver cancer.
New Optical Solitons in Bragg Grating Fibers for the Nonlinear Coupled (
2
+
1
)-Dimensional Kundu-Mukherjee-Naskar System via Complete Discrimination System Method
Authors: Peng Chen , Zhang Furong , Zhao Hongwei , Li Zhao
Published: June 2022

In this paper, the coupled Kundu-Mukherjee-Naskar (KMN) model in Bragg grating fibers is considered to retrieve some new optical soliton solutions in (

2
+
1

) dimensions. Plenty of new exact solutions, including rational function solutions and triangle function solutions, in addition to the Jacobian elliptic function solutions, are obtained by using a complete discrimination system method. The 3D-surface plots, 2D-shape plots, and corresponding 2D contour plots of some obtained solutions are drawn, which provides a visualized structures and propagation of solitons. The novel results are new and show the effectiveness of the proposed method.
Schur, Hermite-Hadamard, and Fejér Type Inequalities for the Class of Higher-Order Generalized Convex Functions
Authors: Ma Yi , Saleem Muhammad Shoaib , Bashir Imran , Xiao Yeliang
Published: June 2022

The study of convex functions is an interesting area of research due to its huge applications in pure and applied mathematics special in optimization theory. The aim of this paper is to introduce and study a more generalized class of convex functions. We established Schur (S), Hermite-Hadamard (HH), and Fejér (F) type inequalities for introduced class of convex functions. The results presented in this paper extend and generalize many existing results of the literature.
A NMR‐Based Metabolomic Approach to Investigate the Antitumor Effects of the Novel [Pt(η1‐C2H4OMe)(DMSO)(phen)]+ (phen = 1,10‐Phenanthroline) Compound on Neuroblastoma Cancer Cells
Authors: De Castro Federica , Stefàno Erika , De Luca Erik , Muscella Antonella , Marsigliante Santo , Benedetti Michele , Fanizzi Francesco Paolo
Published: June 2022

NMR‐based metabolomics is a very effective tool to assess the tumor response to drugs by providing insights for their mode of action. Recently, a novel Pt(II) complex, [Pt(ƞ1‐C2H4OMe)(DMSO)(phen)]+ (phen =  1,10‐phenanthroline), Pt‐EtOMeSOphen, was synthesized and studied for its antitumor activity against eight human cancer cell lines. Pt‐EtOMeSOphen showed higher cytotoxic effects than cisplatin in most of the cancer cell lines and in particular against the neuroblastoma cell line (SH‐SY5Y). In this study, the mechanism of action of Pt‐EtOMeSOphen on SH‐SY5Y cells was investigated using 1H NMR‐based metabolomics and compared with cisplatin. The observed time response of SH‐SY5Y cells under treatment revealed a faster action of Pt‐EtOMeSOphen compared with cisplatin, with a response already observed after six hours of exposure, suggesting a cytosolic target. NMR‐based metabolomics demonstrated a peculiar alteration of the glutathione metabolism pathway and the diacylglycerol expression.
Research on Machine Learning Algorithm for Internet of Things Information Security Management System Research and Implementation
Authors: Jiang Weixiang
Published: June 2022

To study in the Internet information security and privacy problem, a method based on IoT card monitoring technology based on machine learning, the technology can use fuzzy

c

-means algorithm for online business audit, using the Naive Bayes algorithm to classify Internet content and text messages, on the analysis of the comparison of the measurement method based on distance and based on the similarity. The concept of divergence in information theory is used to measure the difference between probability distributions. Finally, the feasibility of the proposed method is validated using the data from the machine learning database. The experimental results show that under the same privacy gain, the data availability relationship between the three anonymous protection methods is roughly satisfied as follows: when the privacy gain is less than 0.85,

U

dverity

<

U

emonymity

<

U

eclorenes

; when the privacy gain is greater than 0.85,

U

uxanaymiy

<

U

uiversriy

<

U

closencs

. And it proves that on the premise of ensuring efficiency and accuracy, the system can find a large number of illegal IoT cards and effectively guarantee the security of IoT.
Building Information Protection Method of Urban Historical Features Based on BIM Technology
Authors: Chen Lin , Chen Xiaolong , Lang Lingyun
Published: June 2022

In order to improve the building information protection method of urban historical features, this paper combines BIM technology to study the building information protection method of urban historical features and studies the least-squares fitting technology of NURBS curves. On this basis, this paper proposes a fitting method of the constrained least-squares method. Moreover, this paper designs a constrained least-squares method for the requirement of fitting accuracy and gives a method for setting the initial value. In addition, this paper proposes a NURBS interpolation method based on arc length parameter compensation technology. The experimental study verifies that the building information protection method of urban historical features based on BIM technology has a good effect and can effectively promote the building information protection of urban historical features.
RecIoT: A Deep Insight into IoT-Based Smart Recommender Systems
Authors: Sharma Richa , Rani Shalli , Nuagh Stephen Jeswinde
Published: June 2022

Typically, Internet of Things (IoT) is perceived as the technology for connecting people, devices, vehicles, home appliances, etc., and 212 billion heterogenous devices are presumed to be connected by 2020 under its umbrella. This domain is gaining popularity worldwide because of the wave it has brought in both research and industrial sector. Smart applications of this scenario provide assistance and services to the users in smart homes, smart cities, smart retail, agriculture, healthcare, etc. However, researchers are now exploring the potential of IoT systems in other domains, one of them being recommender systems. In recent years, the focus of researchers has drifted towards utilizing recommender systems to improve offered selections to IoT users. Therefore, the aim of this paper is to study the present practices, approaches, and challenges in the aforementioned domain along with possible solutions. Briefly, the contributions of this paper are as follows: (1) providing an overview of the prevailing scenario in a range of application domains associated with IoT based recommender systems, (2) providing a framework of some real-world applications, (3) outlining key areas for the improvement towards context-aware personalized services in the future research, and (4) providing a bibliometric trend analysis prevalent in the concerned domain.
Boundary Value Problems for Liénard-Type Equations with Quadratic Dependence on the “Velocity”
Authors: Kirichuka A. , Sadyrbaev F.
Published: June 2022

The estimates were obtained for the number of solutions for the Neumann and Dirichlet boundary value problems associated with the Liénard equation with a quadratic dependence on the “velocity.” Sabatini’s transformation is used to reduce this equation to a conservative one, which does not contain the derivative of an unknown function. Despite the one-to-one correspondence between the equilibria, the topological structure of the phase portraits of both equations can differ significantly.
Application of an Anaerobic–Anoxic–Oxic–Oxic (AAO/O) Model to the Treatment of Real Domestic Wastewater
Authors: Vo Duc-Thuong , Phan Hoang-Vinh-Truong , Hoang Le-Thuy-Thuy-Trang , Nguyen Van-Kieu , Tran Thanh-Nha , Dao Minh-Trung
Published: June 2022

Untreated or inadequately treated domestic wastewater has adversely affected the aquatic environment and public health in many cities in Vietnam. A conventional anaerobic–anoxic–oxic (AAO) process is recognized as an easy-to-handle approach that constrains chemical use during the procedure. Herein, we improve an AAO system by adding more oxic orders in association with a biological membrane in order to increase the hydraulic retention time (HRT) of the oxic zone in the system. The investigated system was applied to the treatment of real domestic wastewater during 168 days of operation. The performance of the system reached a stable state after 60 days of operation. The removal efficiency of total nitrogen (TN), total phosphorus (TP), total suspended solids (TSS), biological oxygen demand (BOD5), and chemical oxygen demand (COD) was found to be 93.6 ± 3.0%, 91.9 ± 3.5%, 88.6 ± 1.2%, 82.6 ± 1.4%, and 71.8 ± 0.7%, respectively. After the operation process, the TN, TP, and TSS contents in the wastewater effluents met the A level in accordance with the QCVN 14-MT:2015/BTNMT regulation, and the effluents of COD and BOD5 almost satisfied the requirement, with only some points being slightly higher than the limit values. The obtained data revealed that the AAO/O system was capable of treating domestic wastewater in small and medium-sized domestic wastewater treatment facilities.
Environmental Affection‐Driven English Tense Analysis: A Healthcare Exercise‐Based Corpus Case Study over Public English Environment
Authors: Ding Yiling , Wang Tianhua
Published: June 2022

Most international academic papers are written in English, and the use of tenses in English academic papers often follows some conventional rules. Automatically extracting and analyzing English tenses in scientific papers have begun to attract researchers’ attention for the global environment. In the analysis of the English tense of scientific papers, consider that the neural network model that combines attention mechanism and sequential input network such as Long Short‐Term Memory (LSTM) network has a long training time, low extraction accuracy, and cannot parallelize text input. We propose an environmental affection‐driven English tense analysis model, which includes an attention mechanism and LSTM model and conducts a temporal analysis of English texts based on an affective computing model. In this paper, our proposed method is verified based on the self‐built healthcare exercise‐based corpus over public English environment. By comparison, the experimental results show that the method proposed in this paper has better performance than ordinary Convolutional Neural Network (CNN), Support Vector Machine (SVM), and LSTM based on attention mechanism.
Expression of 11HOXGenes Is Deregulated in Esophageal Squamous Cell Carcinoma
Authors: Chen Ke-Neng , Gu Zhen-Dong , Ke Yang , Li Ji-You , Shi Xiao-Tian , Xu Guang-Wei
Published: June 2022

Purpose: HOX genes are vital for all aspects of mammalian growth and differentiation, and recent data have shown that their deregulated expression is related to carcinogenesis. To date, there has been no systemic study on expression of HOX genes in esophageal carcinoma. We investigated the expression pattern of 39 known HOX genes in cancerous and noncancerous tissue from 36 patients with esophageal squamous cell carcinoma to determine whether their expression is altered in esophageal cancer.

Experimental Design: Thirty-six patients with resectable esophageal squamous cell carcinoma (ESCC) were enrolled in this study. Specific primers were designed for each of 39 HOX genes, and reverse transcription-PCR was done in cancerous and noncancerous samples of these 36 patients. Furthermore, the expression of HOXA9 protein was subjected to Western blot analysis in all 36 paired tissue samples.

Results: Eight of 39 HOX genes were expressed in cancerous but not in noncancerous tissue. Five of 39 HOX genes were expressed both in cancerous and noncancerous tissue. Of the latter, expression of HOXA7, HOXA9, and HOXC6 was significantly higher in cancerous tissue (P < 0.05). The remaining 26 HOX genes were not detected in either types of tissue. HOXA9 protein was expressed in both kinds of tissue (cancer tissue versus noncancerous mucosa: 0.34 ± 0.32 versus 0.24 ± 0.27, P = 0.121).

Conclusions: This is the first comprehensive survey of 39 HOX gene expression in ESCC and noncancerous mucosa. Five of the 39 HOX genes were expressed in both types of tissue indicating their possible role in maintaining normal structure and function of adult esophageal mucosa. Eleven of the 39 HOX genes were deregulated in cancer tissue. These genes possibly participate in the carcinogenesis of ESCC.
Coexpression of Erythropoietin and Erythropoietin Receptor in Von Hippel-Lindau Disease–Associated Renal Cysts and Renal Cell Carcinoma
Authors: Lee Youn-Soo , Vortmeyer Alexander O. , Lubensky Irina A. , Vogel Timothy W.A. , Ikejiri Barbara , Ferlicot Sophie , Benoît Gérard , Giraud Sophie , Oldfield Edward H. , Linehan W. Marston , Teh Bin T. , Richard Stéphane , Zhuang Zhengping
Published: June 2022

Von Hippel-Lindau (VHL) disease is characterized by multiple tumors in specific target organs. The tumors at different sites share distinct morphologic and genetic characteristics but their cell of origin is unknown. We show that VHL disease–associated renal clear cell carcinomas (RCC) consistently coexpress erythropoietin (Epo) and Epo receptor (EpoR). In addition, coexpression of Epo and EpoR is detected in many renal cysts, providing further evidence that renal cysts are potential precursors for RCC. In conjunction with VHL gene deficiency, coexpression of Epo and EpoR in renal cysts and tumors may reflect a developmental arrest in immature mesenchymal cells. Such arrest may lead to autocrine stimulation, cell proliferation, and renal tumor development, similar to tumorigenesis of VHL disease–associated hemangioblastomas.
Molecular Staging of Lymph Nodes from Patients with Esophageal Adenocarcinoma
Authors: Xi Liqiang , Luketich James D. , Raja Siva , Gooding William E. , Litle Virginia R. , Coello Michael C. , Finkelstein Sydney D. , Chestney Melissa L. , Landreneau Rodney J. , Hughes Steven J. , Godfrey Tony E.
Published: June 2022

Purpose: This study was designed to evaluate molecular markers for the detection of micrometastasis in esophageal adenocarcinoma, define algorithms to distinguish positive from benign lymph nodes and to validate these findings in an independent tissue set and in patients with pN0 esophageal adenocarcinoma.

Experimental Design: Potential markers were identified through literature and database searches. All markers were analyzed by quantitative reverse transcription (QRT)-PCR on a limited set of primary tumors and benign lymph nodes. Selected markers were further evaluated on a larger tissue set and classification algorithms were generated for individual markers and combinations. Algorithms were statistically validated internally as well as externally on an independent set of lymph nodes. Selected markers were then used to identify occult disease in lymph nodes from 34 patients with pN0 esophageal adenocarcinoma.

Results: Thirty-nine markers were evaluated, six underwent further analysis and five were analyzed in the external validation study. Two markers provided perfect classification in both the screening and validation sets, although parametric bootstrap analysis estimated 2% to 3% optimism in the observed classification accuracy. Several marker combinations also gave perfect classification in the observed data sets, and estimates of optimism were lower, implying more robust classification than with individual markers alone. Five of thirty-four patients with esophageal adenocarcinoma had positive nodes by multimarker QRT-PCR analysis and disease-free survival was significantly worse in these patients (P = 0.0023).

Conclusions: We have identified novel QRT-PCR markers for the detection of occult lymph node disease in patients with esophageal adenocarcinoma. The objective nature of QRT-PCR results, and the ability to detect occult metastases, make this an attractive alternative to routine pathology.
Prevalence of CD44+/CD24−/low Cells in Breast Cancer May Not Be Associated with Clinical Outcome but May Favor Distant Metastasis
Authors: Abraham Benny K. , Fritz Peter , McClellan Monika , Hauptvogel Petra , Athelogou Maria , Brauch Hiltrud
Published: June 2022

Purpose: Breast cancer is composed of phenotypically diverse populations of cancer cells. The ability to form breast tumors has been shown by in vitro/in vivo studies to be restricted to epithelial tumor cells with CD44+/CD24−/low characteristics. Validation of these findings with respect to detection in clinical samples, prognosis, and clinical relevance is in demand.

Experimental Design: We investigated breast cancer tissues for the prevalence of CD44+/CD24−/low tumor cells and their prognostic value. The study included paraffin-embedded tissues of 136 patients with and without recurrences. In addition, a breast cancer progression array with normal, carcinoma in situ, and carcinoma tissues was analyzed. We applied double-staining immunohistochemistry for the detection of CD44+/CD24−/low cells. Evaluation was by microscopic pathologic inspection and automated image analysis.

Results: CD44+/CD24−/low cells ranged from 0% to 40% in normal breast and from 0% to 80% in breast tumor tissues. The prevalence of CD44+/CD24−/low tumor cells in 122 tumors was ≤10% in the majority (78%) of cases and >10% in the remainder. There was no significant correlation between CD44+/CD24−/low tumor cell prevalence and tumor progression. Although recurrences of tumors with high percentages of CD44+/CD24−/low tumor cells were mainly distant, preferably osseous metastasis, there was no correlation with the event-free and overall survival. There was no influence on the response to different treatment modalities.

Conclusions: Our findings suggest that the prevalence of CD44+/CD24−/low tumor cells in breast cancer may not be associated with clinical outcome and survival but may favor distant metastasis.
Expression of MDA-7/IL-24 and Its Clinical Significance in Resected Non–Small Cell Lung Cancer
Authors: Ishikawa Shinya , Nakagawa Tatsuo , Miyahara Ryo , Kawano Yozo , Takenaka Kazumasa , Yanagihara Kazuhiro , Otake Yosuke , Katakura Hiromichi , Wada Hiromi , Tanaka Fumihiro
Published: June 2022

Purpose: The melanoma differentiation-associated gene-7 (MDA-7) protein, also known as interleukin (IL)-24, is a novel candidate of tumor suppressor that can induce apoptosis experimentally in a variety of human malignant cells including lung cancer cells. However, only one clinical study has documented that MDA-7/IL-24 expression is down-regulated with progression of melanoma. Thus, the present study was conducted to assess the clinical significance of MDA-7/IL-24 expression in non–small cell lung cancer.

Experimental Design: A total of 183 consecutive patients with resected pathologic stage I-IIIA, non–small cell lung cancer were retrospectively reviewed, and immunohistochemical staining was used to detect MDA-7/IL-24 expression.

Results: MDA-7/IL-24 expression was high in 97 (53.0%) patients and low in the other patients. There was no significant correlation between MDA-7/IL-24 status and any patients' characteristic including pathologic stage. There was no significant difference in tumor angiogenesis or proliferative activity according to MDA-7/IL-24 status, but MDA-7/IL-24-high adenocarcinoma showed a significantly higher incidence of apoptotic tumor cell death than MDA-7/IL-24-low adenocarcinoma. MDA-7/IL-24-high patients seemed to show a favorable postoperative prognosis as compared with MDA-7/IL-24-low patients (5-year survival rates, 75.9% and 62.0%, respectively), although the difference did not reach a statistical significance (P = 0.061). Subset analyses showed that positive MDA-7/IL-24 expression was a significant factor to predict a favorable prognosis in adenocarcinoma (P = 0.033), which was confirmed by a multivariate analysis; there was no difference in the prognosis according to MDA-7/IL-24 status in squamous cell carcinoma.

Conclusions: MDA-7/IL-24 status was a significant prognostic factor in lung adenocarcinoma, not in lung squamous cell carcinoma.
The Evolution of Antibodies into Versatile Tumor-Targeting Agents
Authors: Lin Michael Z. , Teitell Michael A. , Schiller Gary J.
Published: June 2022

In recent years, monoclonal antibodies have become important weapons in the arsenal of anticancer drugs, and in select cases are now the drugs of choice due to their favorable toxicity profiles. Originally developed to confer passive immunity against tumor-specific antigens, clinical uses of monoclonal antibodies are expanding to include growth factor sequestration, signal transduction modulation, and tumor-specific drug delivery. In this review, we shall present the origins of antibody therapeutics within the field of immunotherapy and their evolution into effective anticancer agents, then discuss their multiple mechanisms of action, the basis of their tumor selectivity, and their therapeutic properties compared with traditional therapies. Antibodies are complex molecules whose efficacy and toxicity depend on the antigen, the antibody, any conjugated groups, and even the patient. Finally, we shall present new technologies being developed to increase the efficacy and selectivity of antibody-based therapeutics. Interestingly, many of the new approaches straddle the middle ground between immunotherapy and the traditional modalities of chemotherapy and radiotherapy, and can be seen as ways of combining the selectivity of the former with the efficacy of the latter.
Human Monocyte-Derived Dendritic Cells Pulsed with Wild-Type p53 Protein Efficiently Induce CTLs against p53 Overexpressing Human Cancer Cells
Authors: Tokunaga Naoyuki , Murakami Takayoshi , Endo Yoshikatsu , Nishizaki Masahiko , Kagawa Shunsuke , Tanaka Noriaki , Fujiwara Toshiyoshi
Published: June 2022

Purpose: Dendritic cells are the most potent antigen-presenting cells for initiating cellular immune responses. Dendritic cells are attractive immunoregulatory cells for cancer immunotherapy, and their efficacy has been investigated in clinical trials. The tumor suppressor gene p53 is pivotal in the regulation of apoptosis, and p53-based immunization is an attractive approach to cancer immunotherapy because of the accumulation of p53 protein in malignant but not in normal cells. It has been shown that dendritic cells transduced with an adenoviral wild-type p53 (wt-p53) construct mediate the antitumor immune responses against p53-overexpressing tumor cells. We examined whether monocyte-derived human dendritic cells pulsed with the purified full-length wt-p53 protein were also capable of inducing the specific antitumor responses against p53-overexpressing tumors in vitro.

Experimental Design: Immature dendritic cells generated in the presence of interleukin-4 and granulocyte/macrophage colony-stimulating factor from monocytes of HLA-A2- or HLA-A24-positive healthy individuals were pulsed with the purified p53 protein. Uptake of p53 protein by human dendritic cells was assessed by Western blotting and immunohistochemical staining using anti-p53 antibody. Induction of p53-specific CTL response was also evaluated by the cytotoxic assay against p53-overexpressing human tumor cells.

Results: Both Western blot and immunohistochemical analysis showed the accumulation of p53 protein in human immature dendritic cells. T cells obtained from HLA-A2- or HLA-A24-positive healthy donors were stimulated twice with p53 protein-pulsed dendritic cells and then applied to the cytotoxicity assay against p53-overexpressing target cells. The CTL activity was specific for p53-overexpressing tumor cells and MHC class I restricted. Moreover, the CTL activity generated by p53 protein-pulsed dendritic cells was nearly identical with that induced by adenoviral wt-p53-transduced dendritic cells.

Conclusions: Our results indicate that monocyte-derived human dendritic cells pulsed with the wt-p53 protein could induce the specific antitumor effect against p53-overexpressing tumors and that this in vitro model offers a new and more simple approach to the development of p53-based immunotherapy.
Prognosis of Non–Small Cell Lung Cancer Patients by Detecting Circulating Cancer Cells in the Peripheral Blood with Multiple Marker Genes
Authors: Sher Yuh-Pyng , Shih Jin-Yuan , Yang Pan-Chyr , Roffler Steve R. , Chu Yi-Wen , Wu Cheng-Wen , Yu Chia-Li , Peck Konan
Published: June 2022

Purpose: Current lung cancer staging and prognosis methods are based on imaging methods, which may not be sensitive enough for early and accurate detection of metastasis. This study aims to validate the use of a panel of markers for circulating cancer cell detection to improve the accuracy of cancer staging, prognosis, and as a rapid assessment of therapeutic response.

Experimental Design: We analyzed the National Cancer Institute-Cancer Genome Anatomy Project database to identify potential marker genes for the detection of circulating cancer cells in peripheral blood. Nested real-time quantitative PCR and a scoring method using cancer cell load Lc were employed to correlate the amount of circulating cancer cells with clinical outcomes in 54 non–small cell lung cancer (NSCLC) patients. The Kaplan-Meier method was employed for analysis of prognostic variables.

Results: A panel of four marker genes was identified and experimentally validated. With these marker genes, we achieved an overall positive detection rate of 72% for circulating cancer cells in the peripheral blood of NSCLC patients. Patients who had higher Lc values had worse outcomes and shorter survival times. Patients with poor therapeutic response were revealed by positive detection of circulating cancer cells after therapy. The results correlated well with the patients' survival time.

Conclusion: Circulating cancer cell detection by a panel of markers and the Lc scoring method can supplement the current tumor, node, metastasis staging method for improved prognosis and for rapid assessment of therapeutic response. Together, they may facilitate the design of better therapeutic strategies for the treatment of NSCLC patients.
Erythropoietin and Erythropoietin Receptor Expression in Head and Neck Cancer: Relationship to Tumor Hypoxia
Authors: Arcasoy Murat O. , Amin Khalid , Chou Shu-Chuan , Haroon Zishan A. , Varia Mahesh , Raleigh James A.
Published: June 2022

Purpose: Erythropoietin, an oxygen-regulated glycoprotein hormone, is a hematopoietic cytokine that stimulates erythropoiesis by binding to its cellular receptor [erythropoietin receptor (EPOR)]. The recombinant form of human erythropoietin is used to prevent or treat anemia in cancer patients. However, in a recent randomized, placebo-controlled trial involving patients receiving curative radiotherapy for squamous cell carcinoma of the head and neck, erythropoietin treatment was associated with poorer locoregional progression-free survival. The purpose of our study was to determine whether EPOR and its ligand erythropoietin are expressed in primary head and neck cancer. We also investigated the hypothesis that erythropoietin expression in malignant cells may be associated with the presence of tumor hypoxia, an important factor involved in resistance to radiation treatment, tumor aggressiveness, and poor prognosis.

Experimental Design: Twenty-one patients received an i.v. infusion of the hypoxia marker pimonidazole hydrochloride before multiple tumor biopsies. Contiguous sections from 74 biopsies were analyzed by immunohistochemistry for EPOR and erythropoietin expression and pimonidazole binding.

Results: EPOR expression was present in tumor cells in 97% of the biopsies. Coexpression of erythropoietin was observed in 90% of biopsies. Erythropoietin and pimonidazole adduct staining did not always colocalize within tumors, but there was a significant positive correlation between levels of microregional erythropoietin expression and pimonidazole binding.

Conclusions: The coexpression of erythropoietin and EPOR in tumor cells suggests that erythropoietin may potentially function as an autocrine or paracrine factor in head and neck cancer. The expression of the hypoxia-inducible protein erythropoietin in tumor cells correlates with levels of tumor hypoxia.
Insulin-Like Growth Factor Type 1 Receptor Expression Correlates to Good Prognosis in Highly Malignant Soft Tissue Sarcoma
Authors: Åhlén Jan , Wejde Johan , Brosjö Otte , von Rosen Anette , Weng Wen-Hui , Girnita Leonard , Larsson Olle , Larsson Catharina
Published: June 2022

Purpose: To evaluate known and suggested prognostic markers, especially insulin-like growth factor type 1 receptor (IGF-1R), in highly malignant soft tissue sarcomas (STS).

Experimental Design: A cohort of 101 patients with primary STS of high malignancy grade was studied with respect to development of metastasis, local recurrence, and survival during a minimum of 5 years follow-up. All tumors were analyzed by immunohistochemistry for expression of Ki-67, p53, p27, Bcl-2, IGF-1R, and microvessel density. The traditional clinical variables size, malignancy grade (3 or 4), necrosis, mitotic frequency, infiltrative tumor growth, vascular invasion, depth, and surgical margins were also evaluated.

Results: A significant association was shown between high expression of IGF-1R and favorable outcome. Among STS with positive IGF-1R immunoreactivity, cases with high expression (76-100% positive cells) had the best outcome, whereas cases with the lowest expression (1-25% positive cells) had the worst. As expected, large tumor size (>11 cm), presence of necrosis, high mitotic count, intralesional surgery, and deep location were all significantly associated with poor outcome, both in univariate and multivariate analyses. No difference in outcome was observed between cases of malignancy grade 3 versus 4, whereas the included and more objective variables necrosis and mitotic count were found to be reliable prognostic markers.

Conclusion: IGF-1R expression is a common feature of highly malignant STS. Further elucidation of the role of IGF-1R and the IGF system in STS may both provide a basis for development of new prognostic tools in STS, as well as shed light on the basic mechanisms of the STS development.
Loss of the AP-2α Transcription Factor Is Associated with the Grade of Human Gliomas
Authors: Heimberger Amy B. , McGary Eric C. , Suki Dima , Ruiz Maribelis , Wang Huamin , Fuller Gregory N. , Bar-Eli Menashe
Published: June 2022

Purpose: The activator protein (AP)-2α transcription factor plays a crucial role in the progression of several human tumors, including malignant melanoma, prostate, and breast cancer. Loss of AP-2α results in deregulation of several genes with AP-2α binding motifs such as E-cadherin, p21WAF1, MMP-2, MCAM/MUC18, VEGF, and c-KIT. The purpose of our study was to determine AP-2α expression distribution among grades of gliomas and any possible effect on prognosis.

Experimental Design: A tissue microarray was assembled from all surgical glioma cases with available tissue samples at M.D. Anderson Cancer Center since 1986 to include 72 glioblastomas, 49 anaplastic astrocytomas, 9 low-grade astrocytoma, 37 oligodendrogliomas, 37 anaplastic oligodendrogliomas, 15 mixed oligoastrocytomas, 20 anaplastic mixed oligoastrocytomas, and 7 gliosarcomas. The microarray included normal brain tissue, and AP-2α expression was determined by immunohistochemistry.

Results: AP-2α expression was lost on 99% (P < 0.001) and 98% (P < 0.001) of glioblastomas and anaplastic astrocytomas, respectively, compared with grade 2 astrocytomas and normal brain, all of which (100%) maintained expression of AP-2α. The loss of AP-2α was a negative prognostic indicator within the overall category of gliomas by univariate analysis (rate ratio, 4.30; 95% confidence interval, 2.60-7.10; P < 0.001). However, there was no significant effect of loss of AP-2α expression on survival observed after adjustment for patient age, Karnofsky Performance Scale score, tumor grade, and extent of resection (rate ratio, 1.2; 95% confidence interval, 0.6-2.2; P = 0.6).

Conclusions: AP-2α expression correlates inversely with glioma grade, suggesting a direct role in glioma tumorigenicity, possibly through subsequent deregulation of target genes. Of all the previously characterized markers of progression, the loss of AP-2α would be the most common (96.2%) molecular marker as an astrocytic tumor evolves from grade 2 to 3.
Inhibition of Platelet-Derived Growth Factor Receptor Signaling Restricts the Growth of Human Breast Cancer in the Bone of Nude Mice
Authors: Lev Dina Chelouche , Kim Sun Jin , Onn Amir , Stone Valerie , Nam Do-Hyun , Yazici Sertac , Fidler Isaiah J. , Price Janet E.
Published: June 2022

Purpose: Bone is a common site for breast cancer metastasis. Platelet-derived growth factor (PDGF) and PDGF receptors (PDGFR) are involved in the regulation of bone resorption. This study examined the effects of STI571 (imatinib mesylate), which inhibits PDGFR tyrosine kinase signaling, on the growth of human breast cancer cells in the bone of nude mice with consequent osteolysis.

Experimental Design: Human breast cancer MDA-MB-435 cells were injected into the tibia of female nude mice. Two weeks later the mice were treated with p.o. and injected water (control), daily p.o. STI571, weekly injection of paclitaxel, or daily STI571, plus weekly paclitaxel, for up to 8 weeks. Growth of tumors in bones and osteolysis were monitored by digital radiography and tumors were collected for histochemical analysis.

Results: Mice treated with STI571 or STI571 plus paclitaxel had smaller bone tumors with less lytic bone destruction than did mice treated with water or paclitaxel alone. The results of treatment with paclitaxel plus STI571 did not differ from those with STI571 alone. Immunohistochemistry showed that PDGF-A, PDGF-B, PDGFRα, and PDGFRβ were expressed in the bone tumors. STI571 treatment inhibited PDGFR phosphorylation in tumor cells and tumor-associated endothelial cells, coincident with increased apoptosis, reduced proliferation, and lower microvessel density in the tumors.

Conclusions: Activated PDGFRs are expressed by endothelial and tumor cells in breast cancer tumors growing in the bone of nude mice. Interfering with PDGFR signaling may be an approach to control the progressive growth of breast cancer cells and thus reduce bone lysis.
Growth Suppression of Ovarian Cancer Xenografts in Nude Mice by Vitamin D Analogue EB1089
Authors: Zhang Xiaohui , Jiang Feng , Li Pengfei , Li Chunrong , Ma Qiuping , Nicosia Santo V. , Bai Wenlong
Published: June 2022

Purpose: The poor response of advanced epithelial ovarian cancer to current treatments necessitates the development of alternative therapeutic strategies. Inhibition of cancer growth by 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] compounds represents an innovative approach for cancer therapy. The current study evaluated the therapeutic potential of a synthetic 1,25(OH)2D3 analogue EB1089 in the treatment of ovarian cancer.

Experimental Design: The response of human ovarian cancer cells to 1,25(OH)2D3 and EB1089 were first compared in cell growth, gene transcription, and apoptotic assays. Then, nude mice bearing OVCAR3 tumor xenografts were treated with EB1089 at different dosages, and tumor volumes were monitored. The effect of EB1089 and 1,25(OH)2D3 on the level of serum calcium was also examined. After the treatment, tumors were excised and processed for histologic examination, Ki-67 staining, and tissue terminal deoxynucleotide transferase–mediated dUTP nick end labeling (TUNEL) assays to evaluate the morphologic, proliferative, and apoptotic changes induced by EB1089, respectively.

Results: The study shows that EB1089 suppresses the in vitro growth of ovarian cancer cells and transcriptionally activates the GADD45 reporter gene more effectively than 1,25(OH)2D3. Clinically more importantly, EB1089 suppresses the growth of OVCAR3 tumor xenografts in nude mice without inducing hypercalcemia. Ki-67 staining and tissue TUNEL assays showed that both inhibition of cell proliferation and induction of apoptosis contribute to the EB1089-induced tumor suppression in vivo.

Conclusions: This study is the first demonstration that ovarian cancer responds positively in vivo to treatment with a 1,25(OH)2D3 compound and thus supports continued development of 1,25(OH)2D3 analogues for possible use as an alternative or complementary therapy for human ovarian cancer.
Herstatin, an Autoinhibitor of the Epidermal Growth Factor (EGF) Receptor Family, Blocks the Intracranial Growth of Glioblastoma
Authors: Staverosky Julia A. , Muldoon Leslie L. , Guo Shuhua , Evans Adam J. , Neuwelt Edward A. , Clinton Gail M.
Published: June 2022

Purpose: Herstatin, an autoinhibitor of the epidermal growth factor (EGF) receptor family, was evaluated for efficacy against human glioblastoma in vitro and in vivo in a rat intracranial model.

Experimental Design: Glioblastoma controlled by EGF receptor (EGFR; U87MG) or by the truncated mutant, ΔEGFR (U87MG/Δ), were transfected with Herstatin and evaluated for in vitro and in vivo growth in nude rat brain. Cells treated with purified Herstatin in vitro were evaluated for growth and signal transduction.

Results: Herstatin expression prevented tumor formation by U87MG and purified Herstatin inhibited their growth in vitro in a dose-responsive fashion, whereas in vivo and in vitro growth of U87MG/Δ was resistant to Herstatin. Inhibition of U87MG growth correlated with suppressed EGF activation of EGFR and of Akt but not mitogen-activated protein kinase signaling pathways, whereas ΔEGFR activity and intracellular signaling in U87MG/Δ were unaffected by Herstatin treatment.

Conclusions: Herstatin may have utility against glioblastoma driven by the EGFR but not the mutant ΔEGFR. Blockade of Akt but not the mitogen-activated protein kinase signaling cascade appears to be critical for suppression of intracranial tumor growth.
A Novel Assay to Assess Primary Human Cancer Infectibility by Replication-Selective Oncolytic Adenoviruses
Authors: Wang Yaohe , Thorne Stephen , Hannock Joseph , Francis Jennelle , Au Tina , Reid Tony , Lemoine Nick , Kirn David , Halldén Gunnel
Published: June 2022

Purpose: Replication-selective oncolytic adenoviruses hold promise for cancer treatment, but the predictive use of cell lines, dissociated tumor tissue, and animal models for efficacy against primary cancers are unclear. To further evaluate cytotoxicity and the potential for efficacy of replication-competent adenoviruses we therefore developed a novel methodology using primary human cancer specimens ex vivo; ovarian, colon, rectal, and breast carcinomas were included.

Experimental Design: Tissue culture conditions were developed to maintain viability of adenocarcinomas ex vivo for 48 hours postsurgery. Explants were infected by replication-competent (wild type 5 and E1A mutant dl922-947) and replication-defective (dl312) adenoviruses; early (E1A) and late (hexon) viral gene expression, αv integrins, coxsackievirus and adenovirus receptor (CAR) and tissue viability were assessed by immunohistochemistry and histopathology. Viral replication was verified by replication assays on selected samples.

Results: Viral gene expression varied dramatically among cancer specimens (n = 41). With Ad5, hexon expression was high in 8 of 11 tested specimens, whereas E1A levels were detectable in 16 of 27 tumor explants. Viral gene expression, distribution, and cytopathic effects were greater postinfection with dl922-947. Specimens that supported early gene expression (E1A) also supported viral replication in 13 of 14 tested cases, determined by recovery of infectious units. As predicted, the replication-defective adenovirus dl312 was not associated with viral gene expression.

Conclusions: Primary human tumor tissue remained viable when cultured ex vivo enabling evaluation of viral mutants in tissue with intact morphology. This assay may have great use in determining treatment-sensitive cancers and assess specific oncolytic mutants in individual cases.