bluuebunny/crossref_metadata_2025 huggingface dataset:
A Computational Drug-Target Network for Yuanhu Zhitong Prescription
Authors: Xu Haiyu , Tao Ye , Lu Peng , Wang Peng , Zhang Fangbo , Yuan Yuan , Wang Songsong , Xiao Xuefeng , Yang Hongjun , Huang Luqi
Published: May 2013

Yuanhu Zhitong prescription (YZP) is a typical and relatively simple traditional Chinese medicine (TCM), widely used in the clinical treatment of headache, gastralgia, and dysmenorrhea. However, the underlying molecular mechanism of action of YZP is not clear. In this study, based on the previous chemical and metabolite analysis, a complex approach including the prediction of the structure of metabolite, high-throughputin silicoscreening, and network reconstruction and analysis was developed to obtain a computational drug-target network for YZP. This was followed by a functional and pathway analysis by ClueGO to determine some of the pharmacologic activities. Further, two new pharmacologic actions, antidepressant and antianxiety, of YZP were validated by animal experiments using zebrafish and mice models. The forced swimming test and the tail suspension test demonstrated that YZP at the doses of 4 mg/kg and 8 mg/kg had better antidepressive activity when compared with the control group. The anxiolytic activity experiment showed that YZP at the doses of 100 mg/L, 150 mg/L, and 200 mg/L had significant decrease in diving compared to controls. These results not only shed light on the better understanding of the molecular mechanisms of YZP for curing diseases, but also provide some evidence for exploring the classic TCM formulas for new clinical application.
Interrater Reliability of Chinese Medicine Diagnosis in People with Prediabetes
Authors: Grant Suzanne J. , Schnyer Rosa N. , Chang Dennis Hsu-Tung , Fahey Paul , Bensoussan Alan
Published: May 2013

Background. Achieving reproducibility in research design is challenging when patient cohorts under study are inconsistently defined. Traditional Chinese medicine (TCM) diagnosis is one example where inconsistency between practitioners has been found. We hypothesise that the use of a validated instrument may improve consistency. Biochemical biomarkers may also be used enhance reliability.Methods. Twenty-seven participants with prediabetes were assessed by two TCM practitioners using a validated instrument (TEAMSI-TCM). Inter-rater reliability was summarised using percentage agreement and the kappa coefficient. One-way ANOVA and Tukey’s post hoc test were used to test links between TCM diagnosis and biomarkers.Results. The two practitioners agreed on primary diagnosis of 70% of participants. kappa = 0.56 (P<0.001). The three predominant TCM diagnostic patterns for people with prediabetes were Yin deficiency, Qi and Yin deficiency and Spleen qi deficiency. The Spleen Qi deficiency with Damp cohort had statistically significant higher fasting glucose, higher insulin, higher insulin resistance, higher HbA1c and lower HDL than those with Qi and Yin deficiency.Conclusions. Using the TEAMSI-TCM resulted in moderate interrater reliability between TCM practitioners. This study provides initial evidence of variation in the biomarkers of people with prediabetes according to the different TCM patterns which may suggest a route to further improving interrater reliability.
Accumulation of Mast Cells in the Lesions and Effects of Antiallergic Drugs on the Patients with Inflammatory Bowel Disease
Authors: Kurosawa Motohiro , Nagai Hiroichi
Published: May 2013

The pathomechanism of inflammatory bowel disease (IBD) has not yet been fully demonstrated. However, it is well known that mast cells are present in the gastrointestinal tract, suggesting that mast cells may take part in it. So, we investigated the number of mast cells in IBD, such as ulcerative colitis (UC) and eosinophilic colitis, and showed that the number of mast cells was increased in the inflammatory lesions. We also presented a case of UC which was treated successfully with an antiallergic drug, tranilast. Furthermore, possible new approaches to treating the disease with immunomodulators including suplatast are introduced. However, our investigations were performed with a limited number of patients with IBD, and additional further studies are required to confirm the findings.
Single-Machine Scheduling with Upper Bounded Maintenance Time under the Deteriorating Effect
Authors: Xue Pengfei , Zhang Yulin
Published: May 2013

We consider a single-machine scheduling problem with upper bounded actual processing time and upper bounded maintenance time under deteriorating effect. The actual processing time of a job is a position-dependent power function. If the actual processing time of a job exceeds the upper bound, tardiness penalty of the job should be paid. And if the maintenance time exceeds the corresponding upper bound, tardiness penalty of the maintenance should also be paid. The maintenance duration studied in the paper is a position-dependent exponential function. The objective is to find jointly the optimal maintenance frequency and the optimal job sequence to minimize the total cost, which is a linear function of the makespan and the total tardiness. We show that the studied scheduling problem can be transformed as a classic assignment problem to solve. There is also shown that a special case of the scheduling problem can be optimally solved by a lower order algorithm.
Purpura Fulminans following Thermal Injury
Authors: Hu Jiongyu , Jiang Xupin , He Ting , Luo Qizhi
Published: May 2013

Purpura fulminans is a rare syndrome of intravascular thrombosis and hemorrhagic infarction of the skin, which is an unusual cutaneous manifestation of disseminated intravascular coagulation. It often occurs in small children and babies due to infection and/or sepsis, rarely in adults in clinic. We report the first case of deadly purpura fulminans following thermal injury in a 64-year-old Chinese woman. The purpura developed sharply and aggravated multiple organ dysfunction. The patient died of purpura fulminans, disseminated intravascular coagulation, and multiple organ dysfunction syndrome.
Isolation and Partial Characterization of Bioactive Fucoxanthin fromHimanthalia elongataBrown Seaweed: A TLC-Based Approach
Authors: Rajauria Gaurav , Abu-Ghannam Nissreen
Published: May 2013

Seaweeds are important sources of carotenoids, and numerous studies have shown the beneficial effects of these pigments on human health. In the present study,Himanthalia elongatabrown seaweed was extracted with a mixture of low polarity solvents, and the crude extract was separated using analytical thin-layer chromatography (TLC). The separated compounds were tested for their potential antioxidant capacity and antimicrobial activity againstListeria monocytogenesbacteria using TLC bioautography approach. For bio-autography, the coloured band on TLC chromatogram was visualized after spraying with DPPH and triphenyl-tetrazolium chloride reagents which screen antioxidant and antimicrobial compounds, respectively, and only one active compound was screened on the TLC plate. Preliminary identification of this active compound was done by comparing its colour andRf(retention factor) value with the authentic fucoxanthin standard. Further, the active compound was purified using preparative TLC. This purified compound showed a strong antioxidant (EC50:14.8±1.27 µg/mL) and antimicrobial (inhibition zone: 10.27 mm, 25 µg compound/disc) activities, which were examined by DPPH scavenging and agar disc-diffusion bioassay, respectively. The bioactivity shown by the purified compound was almost similar to the fucoxanthin standard. The characteristic UV-visible and FT-IR spectra of the purified active compound completely matched with the standard. Hence, the main active compound inH. elongatawas identified as fucoxanthin.
Convexity and Spirallikeness Conditions for Two New General Integral Operators
Authors: Güney H. Özlem , Bulut Serap
Published: May 2013

We define two new general integral operators for certain analytic functions in the unit disc and give some sufficient conditions for these integral operators on some subclasses of analytic functions.
Direct Current Magnetron Glow Discharge Plasma Characteristics Study for Controlled Deposition of Titanium Nitride Thin Film
Authors: Borah Sankar Moni
Published: May 2013

This paper reports on the study of direct current (DC) magnetron glow discharge plasma characteristics in a cylindrical magnetron system in argon-nitrogen. Presence of nitrogen gas makes the plasma environment reactive, and it results in significant changes of the plasma properties—number density, electron temperature, floating potential, and sheath thickness. Applied magnetic field is a parameter which is closely related to proper deposition of thin film. Cylindrical Langmuir probe and Emissive probe are used as diagnostics for the estimation of various plasma parameters indicated earlier. Controlled titanium nitride (TiN) thin film deposition on bell-metal at different argon-nitrogen gases ratio is another important study reported.
Surgical Management of Pediatric Gastroesophageal Reflux Disease
Authors: Jackson Hope T. , Kane Timothy D.
Published: May 2013

Gastroesophageal reflux (GER) is common in the pediatric population. Most cases represent physiologic GER and as the lower esophageal sphincter (LES) matures and a solid diet is introduced, many of these patients (>65%) experience spontaneous resolution of symptoms by two years of age. Those who continue to have symptoms and develop complications such as failure to thrive, secondary respiratory disease, and others are classified as having gastroesophageal reflux disease (GERD). Goals of GERD treatment include the resolution of symptoms and prevention of complications. Treatment options to achieve these goals include dietary or behavioral modifications, pharmacologic intervention, and surgical therapy. This paper will review the clinical presentation of GERD and discuss options for surgical management and outcomes in these patients.
Identification and Characterization of DM1 Patients by a New Diagnostic Certified Assay: Neuromuscular and Cardiac Assessments
Authors: Valaperta Rea , Sansone Valeria , Lombardi Fortunata , Verdelli Chiara , Colombo Alessio , Valisi Massimiliano , Brigonzi Elisa , Costa Elena , Meola Giovanni
Published: May 2013

The expansion of the specific trinucleotide sequence, [CTG], is the molecular pathological mechanism responsible for the clinical manifestations of DM1. Many studies have described different molecular genetic techniques to detect DM1, but as yet there is no data on the analytical performances of techniques used so far in this disease. We therefore developed and validated a molecular method, “Myotonic Dystrophy SB kit,” to better characterize our DM1 population. 113 patients were examined: 20 DM1-positive, 11 DM1/DM2-negative, and13 DM1-negative/DM2-positive, who had a previous molecular diagnosis, while 69 were new cases. This assay correctly identified 113/113 patients, and all were confirmed by different homemade assays. Comparative analysis revealed that the sensitivity and the specificity of the new kit were very high (>99%). Same results were obtained using several extraction procedures and different concentrations of DNA. The distribution of pathologic alleles showed a prevalence of the “classical” form, while of the 96 nonexpanded alleles 19 different allelic types were observed. Cardiac and neuromuscular parameters were used to clinically characterize our patients and support the new genetic analysis. Our findings suggest that this assay appears to be a very robust and reliable molecular test, showing high reproducibility and giving an unambiguous interpretation of results.
Some Results on Fixed and Best Proximity Points of Multivalued Cyclic Self-Mappings with a Partial Order
Authors: De la Sen M.
Published: May 2013

This paper is devoted to investigate the fixed points and best proximity points of multivalued cyclic self-mappings on a set of subsets of complete metric spaces endowed with a partial order under a generalized contractive condition involving a Hausdorff distance. The existence and uniqueness of fixed points of both the cyclic self-mapping and its associate composite self-mappings on each of the subsets are investigated, if the subsets in the cyclic disposal are nonempty, bounded and of nonempty convex intersection. The obtained results are extended to the existence of unique best proximity points in uniformly convex Banach spaces.
Abstract A10: CaMKK2 regulates cellular proliferation and androgen receptor activity during prostate cancer progression
Authors: Karacosta Loukia G. , Foster Barbara A. , Gissou Azabdaftari , Feliciano David M. , Edelman Arthur M.
Published: May 2013

Mounting evidence suggests that the androgen receptor (AR) plays a critical role in prostate cancer (PCa) progression. However, the molecular mechanisms by which the AR regulates cell proliferation in androgen-dependent and castration-resistant PCa are incompletely understood. We report here that calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) expression increases during progression of human PCa and becomes nuclear or perinuclear in advanced disease. In a mouse model of PCa (transgenic adenocarcinoma of mouse prostate, TRAMP), CaMKK2 expression increases with progression and is predominantly nuclear. CaMKK2 expression is markedly higher in the AR-expressing, tumorigenic cell line LNCaP compared to cell lines which are AR-nonexpressing and/or nontumorigenic. Silencing of CaMKK2 expression by RNA interference (RNAi) reduces cell proliferation in LNCaP cells but not in AR-non-expressing PC3 cells. In LNCaP cells, dihydrotestosterone (DHT) induces CaMKK2 mRNA and protein expression while androgen withdrawal suppresses CaMKK2 expression. RNAi of CaMKK2 reduces expression of the AR target, prostate specific antigen (PSA) and the cell cycle regulators cyclin D1 and phospho-pRB. Our results suggest that in PCa progression, CaMKK2 is transcriptionally induced by the AR to maintain AR activity, AR-dependent cell cycle control and continued cell proliferation.

Citation Format: Loukia G. Karacosta, Barbara A. Foster, Azabdaftari Gissou, David M. Feliciano, Arthur M. Edelman. CaMKK2 regulates cellular proliferation and androgen receptor activity during prostate cancer progression [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A10.
Abstract A15: A phase 2 study of neoadjuvant metformin in prostatic carcinoma
Authors: Joshua Anthony Michael , Fleshner Neil , Pollak Michael , Zannella Vanessa , Bowes Barbara , Koritzinsky Marianne , Sweet Joan , Evans Andrew , Trachtenberg John , Jewett Michael , Finelli Antonio
Published: May 2013

Background: Metformin is an inhibitor of the complex 1 in the respiratory chain, and is widely used in diabetes due to its effect on reducing insulin resistance. It has also been recently described to have effects via AMPK on inhibiting the mTOR kinase. Significant preclinical and epidemiological studies suggest its role in chemoprevention. These actions provide significant rationale to evaluate its utility in prostate cancer. We conducted a phase II single centre study of neoadjuvant metformin in localized prostate cancer.

Methods: Men were required to have histologically confirmed prostate cancer involving at least 20% of at least 1 unfragmented biopsy core. Exclusion criteria included patients who on initial assessment are found to be on treatment with any drug used for the treatment of any form of diabetes, or patients that begin treatment for any form of diabetes during the course of the study. Pts were treated with up to 500mg tid of metformin. The primary objectives were to demonstrate safety and tolerability of neoadjuvant metformin administration in men with prostate cancer and to document changes in phospho-AKT signaling indices.

Results: 24 patients were enrolled with 22 evaluable; median age was 64 yrs (range, 45-70 yrs). Baseline characteristics included median PSA 6 ng/mL (range, 3.22–36.11 ng/mL). Median duration of drug treatment was 41 days (range 18–81). No grade 3 adverse events were reported, all patients underwent subsequent radical prostatectomy with adverse effects related to metformin. Significant pre-and post changes were noted in serum IGF1 (p=0.02), fasting glucose (p=0.03), BMI (p < 0.01) and waist/hip ratio (p < 0.01). There was a trend for a PSA reduction (p=0.08). There were no correlations between any metabolic, morphometric or cancer-related serum indices.

Conclusions: Neoadjuvant metformin is well tolerated prior to radical prostatectomy. Data to date indicates promising effects on metabolic parameters, tissue results will be presented including proliferation indices and signaling pathway assessments.

Citation Format: Anthony Michael Joshua, Neil Fleshner, Michael Pollak, Vanessa Zannella, Barbara Bowes, Marianne Koritzinsky, Joan Sweet, Andrew Evans, John Trachtenberg, Michael Jewett, Antonio Finelli. A phase 2 study of neoadjuvant metformin in prostatic carcinoma [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A15.
Abstract A22: Combined simvastatin and anti-IGF-1R therapy of prostate cancer cells in co-culture with bone
Authors: Nordstrand Annika , Lundholm Marie , Larsson Andreas , Lerner Ulf , Widmark Anders , Wikström. Pernilla
Published: May 2013

Prostate cancer frequently metastasizes to the bone. Bone metastases show weak responses to the conventional therapies given today, and patients with metastases cannot be cured. Bone matrix is rich in stored growth factors, with IGF-1 being one of the most abundant. IGF-1 acts as a survival factor for tumor cells and we speculate that bone-derived IGF-1 weakens the effects of therapies aimed to target bone metastases and, furthermore, that effects of apoptosis inducing therapies could be enhanced if given in combination with therapies inhibiting the IGF-1 receptor (IGF-1R). Simvastatin inhibits the mevalonate pathway and have been found to induce apoptosis of prostate cancer cells. The aims of this study were to confirm stimulating effects of bone-derived IGF-1 on prostate cancer cell growth and to test if IGF-1R inhibition could enhance simvastatin therapy effects on prostate cancer cells in a bone environment. Prostate cancer PC-3 and 22Rv1 cells were co-cultured with mice calvarias for 72 h and showed significantly increased cell numbers compared to when cultured alone. A phospho-RTK array confirmed activation of the IGF-1R on PC-3 and 22Rv1 cells by calvaria-conditioned media, and neutralization of bone-derived IGF-1 abolished the calvaria-induced increase of PC-3 cell growth. Treatments of PC-3 and 22Rv1 cells with simvastatin and the IGF-1R inhibitor NVP-AEW541 reduced tumor cell numbers and cell viability, and induced apoptosis, as assessed by flow cytometry analysis of Annexin-V. Combined s imvastatin and NVP-AEW541 treatments resulted in enhanced growth inhibitory effects compared to either therapy given alone. Expression analysis by RT-PCR and Western blot indicated that the therapeutic effects of simvastatin involved down-regulation of the IGF-1R in PC-3 cells and of the constitutively active androgen receptor AR-V7 variant in 22Rv1 cells. In conclusion, we believe that a combination of IGF-1R inhibition with simvastatin, and possibly other apoptosis inducing therapies, can provide novel and improved treatment regimes for prostate cancer bone metastases.

Citation Format: Annika Nordstrand, Marie Lundholm, Andreas Larsson, Ulf Lerner,Anders Widmark, Pernilla Wikström. Combined simvastatin and anti-IGF-1R therapy of prostate cancer cells in co-culture with bone [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A22.
Abstract A24: Epigenetic silencing of ID4 (inhibitor of DNA binding 4) is associated with prostate cancer
Authors: Sharma Pankaj , Chinaranagari Swathi , Patel Divya , Chaudhary Jaideep
Published: May 2013

Introduction: Inhibitor of DNA binding/differentiation protein 4 (Id4) is dominant negative regulator of basic helix loop helix (bHLH) family of transcription factors. Id4 shares the homology of HLH domain with other Ids and lack the basic DNA binding region. Previous studies from our lab had shown that unlike Id1, Id2 and Id3, Id4 acts as a tumor suppressor in prostate cancer by attenuating cell proliferation and promote apoptosis. Consistent with these observations Id4 is epigenetically silenced in Du145 prostate cancer cell line. In this study we investigated whether Id4 is also epigenetically silenced in prostate cancer. We also examined association between Id4 promoter hypermethylation and its expression in prostate cancer cell lines.

Experimental procedures: Id4 protein expression was analyzed on human prostate adenocarcinoma samples by Immunhistochemistry. Id4 promoter methylation pattern on prostate cancer cell lines (C33, C81, DU145, LNCaP, DU145+ID4, and PC3) was examined. In addition, we performed Laser capture microdisection (LCMD) on the human prostate tissues and methylation specific PCR to correlate our cell line studies with clinical studies.

Results: IHC demonstrated decreased Id4 protein expression in human prostate tissue samples whereas higher nuclear Id4 expression was found in normal prostate tissues.Id4 methylation specific PCR (MSP) on prostate cancer cell lines, showed Id4 methylation in DU145 cell and but not in LNCaP and C33 cells. In C81 and PC3 cells it showed partial methylation. C81 are derivatives of LNCaP cell lines and are Androgen independent and are more metastatic. Increased Id4 methylation in C81 as compared to LNCaP suggests its epigenetic silencing as cells acquire androgen independence. Tumors with Id4 promoter hypermethylation showed distinct loss of Id4 expression.

Conclusions: Id4 appears as a potential tumor suppressor gene that is epigenetically silenced during cancer development.Id4 promoter hypermethylation had been reported in breast, colorectal cancer and chronic lymphocytic leukemia Thus, loss of Id4 expression due to promoter hypermethylation. Id4 methylation status could serve as a prognostic biomarker in human prostate cancer.

Acknowledgement: The research was supported by NIH/NCI-RO1CA128914 and in part by NIH/NCRR/RCMI G12RR03062.

Citation Format: Pankaj Sharma, Swathi Chinaranagari, Divya Patel, Jaideep Chaudhary. Epigenetic silencing of ID4 (inhibitor of DNA binding 4) is associated with prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A24.
Abstract A27: Comprehensive analysis of the TMPRSS2-ERG translocation during prostate cancer progression
Authors: Tan Shyh-Han , Sesterhenn I. , Srivastava S. , Bubendorf L. , Ruiz C. , Braun M. , Scheble V.J. , Zellweger T. , Rentsch C.A. , Bachmann A. , Dobi Albert , Perner S.
Published: May 2013

Introduction: Approximately half of the diagnosed prostate carcinomas (PC) are characterized by a chromosomal rearrangement fusing the androgen regulated gene TMPRSS2 to the oncogenic ETS transcription factor ERG. Aim of this study was to comprehensively analyze the impact of this translocation on the expression of the ERG gene in hormone-naïve (untreated) and castration-resistant prostate cancers and to define the influence of AR protein expression and genomic amplification in this context.

Methods: We constructed a tissue microarray (TMA) containing 915 tissue cores from 107 hormone-naïve PC and from 101 castration-resistant (CR) PC. In addition, we included 56 specimens from distant metastases. We analyzed the TMPRSS2-ERG translocation status by fluorescence in-situ hybridization and the expression profiles of ERG, AR and the proliferation marker Ki67 by immunohistochemistry.

Results: Nearly half of the analyzed PC tissue specimens (untreated: 38%, castration-resistant: 46%) harbored a TMPRSS2-ERG gene fusion. Untreated PC with positive translocation status showed increased tumor cell proliferation (p < 0.05). As expected, TMPRSS2-ERG gene fusion was strongly associated with increased ERG protein expression in untreated, as well as in castration-resistant PC (both p < 0.0001). However, we detected a subgroup (26%) of CR PCs with the gene fusion, but without detectable ERG protein expression. This subgroup showed significantly lower levels of AR protein expression and of the androgen regulated serum PSA (both p>0.05).

Conclusions: This comprehensive study comparing the TMPRSS2-ERG gene and ERG protein expression status in a large cohort of hormone-naïve and CR PC, identifies a subgroup of translocated CR PCs without ERG protein expression. Our results suggest that this subgroup may represent CR PCs with a dispensed AR pathway. Further analyses may help to explain if these tumors represent a distinct subgroup of AR independent CR PCs.

Citation Format: Shyh-Han Tan, I. Sesterhenn, S. Srivastava, L. Bubendorf, C. Ruiz, M. Braun, V.J. Scheble, T. Zellweger, C.A. Rentsch, A. Bachmann, Albert Dobi, S. Perner. Comprehensive analysis of the TMPRSS2-ERG translocation during prostate cancer progression [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A27.
Abstract A29: Distinguishing features of ERG oncoprotein expression among matched cohorts of African-American and Caucasian-American prostate cancer patients
Authors: Petrovics Gyorgy , Rosen Philip , Pfister David , Young Denise , Chen Yongmei , Dobi Albert , McLeod David , Srivastava Shiv , Sesterhenn Isabell
Published: May 2013

Introduction and Objective: The ETS-related gene (ERG) proto-oncogene is frequently overexpressed in prostate cancer (CaP) as a result of a genomic rearrangement that places ERG under the control of the androgendependent TMPRSS2 promoter. The initial report of quantitative ERG mRNA expression in micro-dissected prostate tumor cells showed significantly higher ERG expression in CaP of Caucasian-Americans (CA) patients vs. African-American (AA) patients (Petrovics et al, Oncogene, 2005). Recent studies (Magi-Galluzzi et al, Prostate, 2011; Elliott et al, USCAP Mtg., 2011) have also shown higher frequency of ERG rearrangement or ERG oncoprotein expression in CaP of CA patients. We evaluate the frequency and pattern of the ERG oncoprotein expression in prostate tumors of matched CA and AA CaP patients to better understand the biological basis for differences in prostate cancer between the two populations.

Methods: Ninety one AA and 91 CA CaP patients were matched for age, Gleason score and pathologic stage. All underwent radical prostatectomy between 1993 and 2010. Whole mount prostate specimens were used for the immunohistochemical detection of the ERG oncoprotein by a highly specific ERG monoclonal antibody (clone 9FY). Individual foci of tumors were reported as either positive or negative. ERG staining data was linked to clinico-pathologic data. Biochemical recurrence was defined as two serum PSA measurements of 0.2 ng/mL or higher at least 8 weeks from surgery.

Results: A higher percentage of CA (61.8%) than AA (28.2%) CaP patients had ERG positive index tumors and had at least one focus of tumor positive for ERG. A higher percentage of overall tumor foci were also positive for ERG in CA than AA CaP patients. There was high concordance of ERG positive prostatic intra-epitheilal neoplasia (PIN) and ERG positive CaP in both AA and CA patients.

Conclusions: ERG expression is more prevalent among CA than AA CaP patients in matched cohorts. Differences in the pattern of ERG expression in CaP and differing trends in biochemical recurrence between CA and AA patients with ERG (+) index tumors suggest a dominant clonal selection of ERG-positive tumors in CA patients. These differences in ERG expression have the potential to delineate biological distinctions of CaP in the two patient populations.

Citation Format: Gyorgy Petrovics, Philip Rosen, David Pfister, Denise Young, Yongmei Chen, Albert Dobi, David McLeod, Shiv Srivastava, Isabell Sesterhenn. Distinguishing features of ERG oncoprotein expression among matched cohorts of African-American and Caucasian-American prostate cancer patients [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A29.
Abstract A30: ERG protein expression and genomic rearrangement status in primary and metastatic prostate cancer a comparative study of two monoclonal antibodies.
Authors: Sreenath Taduru , Fend Falko , Wernert Nicolas , Kristiansen Glen , Perner Sven , Tan Shyh-Han , Braun Martin , Goltz Diane , Shaikibrahim Zaki , Vogel Wenzel , Boehm Diana , Scheble Veit , Dobi Albert
Published: May 2013

Introduction and Objective: Overexpression of the ERG protein is highly prevalent in prostate cancer (PCa) and most commonly results from gene fusions involving the ERG gene. Recently, an N-terminal epitope targeted mouse and a C-terminal epitope targeted rabbit monoclonal anti-ERG antibody have been introduced for the detection of the ERG protein. Independent studies reported that immunohistochemical (IHC) stains with both monoclonal anti-ERG antibodies (ERG-MAbs) highly correlate with the underlying ERG gene rearrangement status. However, a comparative study of both antibodies has not been provided so far. Here, we are the first to compare the mouse ERG-MAb to the rabbit ERG-MAb for their concordance on the same PCa cohort. Furthermore, we assessed if the ERG protein expression is conserved in lymph node and distant PCa metastases, of which a subset underwent decalcification.

Methods: We evaluated tissue microarrays of 278 specimens containing 265 localized PCa, 29 lymph node, 30 distant metastases, and 13 normal prostatic tissues. We correlated the ERG protein expression with the ERG rearrangement status using an ERG break-apart fluorescence in-situ hybridization (FISH) assay and IHC of both ERG antibodies.

Results: ERG protein expression and ERG rearrangement status were highly concordant regardless of whether the mouse or rabbit ERG-MAb was used (97.8% versus 98.6%, respectively). Of interest, both ERG antibodies reliably detected the ERG expression in lymph node and distant PCa metastases, of which a subset underwent decalcification. If an ERG protein expression was present in localized PCa, we observed the same pattern in the corresponding lymph node metastases.

Conclusions: This is the first study to comprehensively compare the two available ERG-MAbs. By demonstrating a broad applicability of IHC to study ERG protein expression using either antibody, this study adds an important step towards a facilitated routine clinical application. Further, we demonstrate that the clonal nature of the ERG rearrangement is not restricted to the genomic level, but proceeds in the proteome. Together, our results simplify future efforts to further elucidate the biological role of ERG in PCa.

Citation Format: Taduru Sreenath, Falko Fend, Nicolas Wernert, Glen Kristiansen, Sven Perner, Shyh-Han Tan, Sr., Martin Braun, Diane Goltz, Zaki Shaikibrahim, Wenzel Vogel, Diana Boehm, Veit Scheble, Albert Dobi. ERG protein expression and genomic rearrangement status in primary and metastatic prostate cancer a comparative study of two monoclonal antibodies [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A30.
Abstract A31: ERG-mediated alterations in chromatin conformation
Authors: Rickman David S. , Melnick Ari , Elemento Olivier , Rubin Mark , Soong T. David , Moss Benjamin , Mosquera Juan Miguel , Dlabal Jan , Terry Stephane , MacDonald Theresa , Bunting Karen , Demichelis Francesca
Published: May 2013

Emerging evidence suggests that chromatin adopts a non-random three dimensional (3D) topology and that the organization of genes into structural hubs and domains affects their transcriptional status. How chromatin conformation changes in diseases such as cancer is poorly understood. Moreover, how oncogenic transcription factors, which bind to thousands of sites across the genome, influence gene regulation by globally altering the topology of chromatin requires further elucidation. To address these questions, we performed unbiased high-resolution mapping of intra- and inter-chromosome interactions upon over-expression of ERG, an oncogenic transcription factor frequently over-expressed in prostate cancer as a result of a gene fusion. By integrating data from genome-wide chromosome conformation capture (Hi-C), ERG binding (ChIP-seq), gene expression (RNA-seq) and spectral karytopying, we demonstrate for the first time that the oncogenic transcription factor ERG can induce global, reproducible and functionally coherent changes in chromatin organization that are associated with gene expression differences and genomic alterations. The results presented here have broader implications, as many driving genetic lesions in other cancer types involve altered levels of transcription factors due to genomic alterations (e.g. EWS-FLI1, c-Myc, n-Myc, PML-RARα).

Citation Format: David S. Rickman, Ari Melnick, Olivier Elemento, Mark Rubin, T. David Soong, Benjamin Moss, Juan Miguel Mosquera, Jan Dlabal, Stephane Terry, Theresa MacDonald, Karen Bunting, Francesca Demichelis. ERG-mediated alterations in chromatin conformation [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A31.
Abstract A33: Characterization of signaling pathways associated with docetaxel resistance in prostate cancer
Authors: Lee Brian Yong , Hochgrafe Falko , Raftery Mark , Horvath Lisa G. , Daly Roger J.
Published: May 2013

Docetaxel (DTX) is the only treatment option for patients diagnosed with metastatic castration-refractory prostate cancer (CRPC). However, approximately 50% of patients do not respond to DTX, but are exposed to significant toxicity without direct benefit. This study aims to use mass spectrometry (MS) based techniques to characterize the perturbed intracellular signaling, particularly tyrosine phosphorylation, which contributes to the chemoresistant phenotype in CRPC patients, and thereby identify novel therapeutic targets.

Two DTX-resistant prostate cancer cell lines (DU145-Rx & PC3-Rx) were established from their parental DTX-sensitive cell lines, DU145 & PC3, respectively, by a dose-escalation protocol. Phosphotyrosine profiling was performed by immunoaffinity coupled LC-MS/MS in combination with stable isotope labeling with amino acid in cell culture (SILAC). The chemosensitive and resistant cells were cultured in “light” and “heavy” SILAC media, respectively, and their isotopic labels were switched in a replicate experiment. Phosphotyrosine peptides were analysed on the state-of-the-art High Performance Liquid Chromatography MS/MS (Orbitrap Velos ETD). Raw MS spectra were quantified using MaxQuant software.

DU145-Rx & PC3-Rx displayed 44 and 35-fold higher IC50 values, respectively, and significantly greater colony forming ability, in the presence of DTX, compared to the parental lines. Phosphotyrosine profiling of these cell lines identified a total of 365 phosphosites in the chemoresistant lines, corresponding to 227 proteins. 3 kinases and 3 cytoskeletal proteins showed a significant increase in tyrosine phosphorylation in both DTX-resistant models.

These candidate proteins will be further characterized by gain- and loss-of-function approaches, both in vitro and in xenograft models, to elucidate the intracellular mechanism of chemoresistance, and to design therapeutic strategies to reverse the chemoresistant phenotype in CRPC.

Citation Format: Brian Yong Lee, Falko Hochgrafe, Mark Raftery, Lisa G. Horvath, Roger J. Daly. Characterization of signaling pathways associated with docetaxel resistance in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A33.
Abstract A4: Androgen receptor (AR) degradation is controlled by the co-operation of PMEPA1 and the E3 ubiquitin ligase NEDD4-1
Authors: Li Hua , Dobi Albert , Srivastava Shiv
Published: May 2013

Introduction and Objective: Androgen receptor (AR) plays critical roles in prostate growth and differentiation and prostate tumorigenesis. Our recent investigations have discovered a novel mechanism of AR degradation in CaP cells involving PMEPA1 and NEDD4-1 which is independent of MDM2. Previously we have shown that PMEPA1 is a highly androgen-inducible gene in CaP cells that recruits the NEDD4-1, an E3 ubiquitin ligase, to degrade AR. In an independent study NEDD4-1 has been identified as an E3 ligase controlling the levels of PTEN. We hypothesize that decreased or loss of PMPEA1 expression that is frequently observed in human prostate tumor cells may lead to enhanced AR functions. Elevated AR levels may increase NEDD4-1 and in turn destabilize PTEN in prostate cancer cells. Here, we further investigate the co-operation between PMEPA1 and NEDD4-1 to alter AR and PTEN protein levels.

Methods: PMEPA1 and NEDD4-1 were ectopically expressed in androgen responsive LNCaP and VCaP cells by pCMV-XL4 and pcDNA3.1 vectors, respectively. AR, PMEPA1, NEDD4-1, MDM2 and PTEN were silenced by using specific siRNAs. Cell growth was monitored in response modulation of PMEPA1, NEDD4 and AR. Protein levels of AR, PMEPA1, NEDD4-1, PTEN and PSA were assessed by immunoblot assays.

Results: Decreases in AR protein levels, inhibited AR signaling and cell growth in response to heterologous expression of NEDD4-1. Consistent with this observation, inhibition of NEDD4-1 by siRNA resulted in elevated AR protein levels, AR signaling and increased cell growth. Reduced cell growth by AR knock-down or by heterologous expression of NEDD4-, were rescued by the knockdown of PMEPA1. It was also shown that AR up-regulates the expression of NEDD4-1 in hormone responsive CaP cells. In VCaP cells, overexpression of NEDD4-1 resulted in decreased PTEN protein levels. Remarkably, PMEPA1 knockdown resulted in elevated AR and NEDD4-1 levels and the downregulation of PTEN protein.

Conclusions: NEDD4-1 mediated AR degradation in hormone responsive CaP cells requires PMEPA1. The frequently observed decreased expression of PMEPA1 results in elevated AR protein levels and may facilitate NEDD4-1 mediated degradation of PTEN in CaP. Our findings revealed a complex interplay of the AR, PMEPA1, NEDD4 and PTEN underscoring the impact of the loss of PMEPA1 to increased AR and decreased PTEN, both proteins critical in prostate tumorigenesis.

Citation Format: Hua Li, Albert Dobi, Shiv Srivastava. Androgen receptor (AR)degradation is controlled by the cooperation of PMEPA1 and the E3 ubiquitin ligase NEDD4-1 [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A4.
Abstract A41: Effect of dietary vitamin D3 and calcium on the growth of androgeninsensitive human prostate tumor in a murine model
Authors: Ray Rahul , Banks Mara , Abuzahra Hilal , Persons Kelly S. , Eddy Vikram J. , Lambert James R. , Holick Michael F.
Published: May 2013

Vitamin D deficiency has been associated with increased risk of prostate cancer in epidemiologic and prospective studies. An association has also been made between high dietary calcium and increased prostate cancer risk. Previous studies have demonstrated that normal prostate cells and some prostate cancer cells have the ability to convert 25(OH)D to 1α,25(OH)2D. This ability along with epidemiological evidence correlating an inverse relationship between serum 25(OH)D levels and prostate cancer risk suggests that vitamin D may be important for prostate cancer prevention. The effect of dietary vitamin D and calcium on the growth of human androgen-insensitive prostate tumor in an athymic mouse model was evaluated. Three factors were considered in evaluating the role of dietary calcium and vitamin D on prostate tumor in our mice model: tumor-growth, elevation of serum calcium and overall growth of the animals (body weight). Immunohistochemical staining and Real Time PCR analysis were performed on tumors to elucidate the molecular mechanism(s) involved in vitamin D regulation of prostate cancer. Tumor-growth was highest in normal calcium, vitamin D deficient diet group (381±59 mm3) while growth in normal calcium, vitamin D sufficient (179±36 mm3), high calcium, vitamin D sufficient (199±25 mm3) and deficient (204±43 mm3) diets were not significantly different from one another. The normal calcium, vitamin D deficient diet had mice that were normo-calcemic compared to the other diet groups though there was no significant difference between groups for the weights of the animals. Proliferative (Ki67) and apoptotic (Caspase 3) staining revealed no significant difference between vitamin D sufficient and deficient diet groups. Additional staining for p21 did not show any difference among the diet groups. Gene analysis for p21, p27 and p53 did not demonstrate altered transcriptional activity in vitamin D sufficient groups. Bax and bcl-2 mRNA expression were both increased in the high calcium, vitamin D deficient diet group. The difference in tumor growth between the normal calcium, vitamin D deficient diet and the other diets suggest an important role of dietary vitamin D as a preventive agent in androgen-insensitive prostate cancer. More in vivo studies are needed to understand the molecular mechanism(s) behind vitamin D's antitumorigenic effects.

Citation Format: Rahul Ray, Mara Banks, Hilal Abuzahra, Kelly S. Persons, Vikram J. Eddy, James R. Lambert, Michael F. Holick. Effect of dietary vitamin D3 and calcium on the growth of androgeninsensitive human prostate tumor in a murine model [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A41.
Abstract A45: Resveratrol inhibits prostate cancer stem cell characteristics by suppressing Wnt, sonic hedgehog, and Notch pathways
Authors: Srivastava Rakesh K. , Rodova Mariana , Fu Junsheng , Nall Dara , Shankar Sharmila
Published: May 2013

The existence of cancer stem cells (CSCs) in prostate cancer has profound implications for cancer prevention and/or treatment. The CSC population resides within the prostate tumor mass and they may be responsible for drug resistance, and for the recurrence of prostate cancer. Therefore, development of novel strategies to target CSCs by nontoxic agent resveratrol is an innovative and novel idea for the management of prostate cancer. Activation of Wnt, sonic hedgehog (Shh) and Notch pathways are required for the growth of numerous tissues and organs and recent evidence indicates that these pathways are often recruited to regulate self-renewal and metastasis of CSCs. The objectives of this study were to examine the molecular mechanisms by which resveratrol, an active compound in grapes and red vine inhibits self-renewal capacity of prostate CSCs. Our data demonstrated that resveratrol inhibited the expression of pluripotency maintaining transcription factors (Nanog, c-Myc and Oct-4), and spheroid formation by prostate CSCs. Inhibition of Nanog by shRNA enhanced the inhibitory effects of resveratrol on self-renewal capacity of CSCs. Resveratrol suppressed cell proliferation and induced apoptosis by inhibiting the expression of Bcl-2 and IAPs, and activating caspase-3. Interestingly, resveratrol also inhibited the transcriptional activities TCF/LEF, Gli and RBP-Jk. Furthermore, resveratrol inhibited epithelial-mesenchymal transition (EMT) by suppressing the expression of Snail, Slug and ZEB1 transcription factors, which correlated with significantly reduced CSC's migration and invasion, suggesting the blockade of signaling involved in early metastasis. In conclusion, resveratrol can inhibit prostate cancer CSC characteristics by suppressing various signal transduction pathways, and can be used for the management of prostate cancer.

Citation Format: Rakesh K. Srivastava, Mariana Rodova, Junsheng Fu, Dara Nall, Sharmila Shankar. Resveratrol inhibits prostate cancer stem cell characteristics by suppressing Wnt, sonic hedgehog, and Notch pathways [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A45.
Abstract A51: Disruption of Abi1/Hssh3bp1 expression induces prostatic intraepithelial neoplasia in the conditional Abi1/Hssh3bp1 KO mice
Authors: Xiong Xiaoling , Kotula Leszek , Chorzalska Anna , Dubielecka Patrycja , White Julie , Vedvyas Yogindra , Hedvat Cyrus V. , Haimovitz-Friedman Adriana , Koutcher Jason A. , Sawicki Janet A.
Published: May 2013

Prostate cancer is one of the leading causes of cancer-related deaths in the United States and a leading diagnosed non-skin cancer in American men. Genetic mutations underlying prostate cancer tumorigenesis include alterations of tumor suppressor genes. We previously proposed that ABI1/hSSH3BP1 acts as a tumor suppressor gene (Macoska, et al. 2001 Neoplasia 3, 99-104). Here, we tested this hypothesis by searching for ABI1 gene mutations in primary prostate tumors from patients, and by analyzing the consequences of prostate-specific disruption of the mouse Abi1/Hssh3bp1 orthologue. We sequenced the ABI1/hSSH3BP1 gene and identified mutations in 15 out of 44 prostate tumors. Moreover, complementation growth-, proliferation- and xenograft- assays using the LNCaP cell line, which contains the loss-of-function Abi1 mutation, and stably expressed wt ABI gene, were consistent with the tumor suppressor hypothesis. To test the hypothesis further, we disrupted the gene in the mouse prostate by breeding the Abi1 floxed strain with the probasin promoter-driven Cre recombinase strain. Histopathological evaluation of mice indicated development of prostatic intraepithelial neoplasia (PIN) in Abi1/Hssh3bp1 KO mouse as early as the 8th month, but no progression beyond PIN was observed in mice as old as 12 months. Observed decreased levels of E-cadherin, β-catenin, and WAVE2 in mouse prostate suggest abnormal cellular adhesion as the mechanism underlying PIN development due to Abi1 disruption. In summary, this study provides further evidence for the hypothesis that Abi1 down-regulation plays a role in the development of prostate cancer and warrants further investigations of Abi1's role in prostate cancer tumorigenesis.

Citation Format: Xiaoling Xiong, Leszek Kotula, Anna Chorzalska, Patrycja Dubielecka, Julie White, Yogindra Vedvyas, Cyrus V. Hedvat, Adriana Haimovitz-Friedman, Jason A. Koutcher, Janet A. Sawicki. Disruption of Abi1/Hssh3bp1 expression induces prostatic intraepithelial neoplasia in the conditional Abi1/Hssh3bp1 KO mice [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A51.