bluuebunny/crossref_metadata_2025 huggingface dataset:
A Complex Assembly Landscape for the 30S Ribosomal Subunit
Authors: Sykes Michael T. , Williamson James R.
Published: February 2009

The ribosome is a complex macromolecular machine responsible for protein synthesis in the cell. It consists of two subunits, each of which contains both RNA and protein components. Ribosome assembly is subject to intricate regulatory control and is aided by a multitude of assembly factors in vivo, but can also be carried out in vitro. The details of the assembly process remain unknown even in the face of atomic structures of the entire ribosome and after more than three decades of research. Some of the earliest research on ribosome assembly produced the Nomura assembly map of the small subunit, revealing a hierarchy of protein binding dependencies for the 20 proteins involved and suggesting the possibility of a single intermediate. Recent work using a combination of RNA footprinting and pulse-chase quantitative mass spectrometry paints a picture of small subunit assembly as a dynamic and varied landscape, with sequential and hierarchical RNA folding and protein binding events finally converging on complete subunits. Proteins generally lock tightly into place in a 5′ to 3′ direction along the ribosomal RNA, stabilizing transient RNA conformations, while RNA folding and the early stages of protein binding are initiated from multiple locations along the length of the RNA.
The Pathogenesis of Pituitary Tumors
Authors: Asa Sylvia L. , Ezzat Shereen
Published: February 2009

Recently there has been significant progress in our understanding of pituitary development, physiology, and pathology. New information has helped to clarify the classification of pituitary tumors. Epidemiologic analyses have identified a much higher incidence of pituitary tumors than previously thought. We review the pathogenetic factors that have been implicated in pituitary tumorigenesis and the application of novel targeted therapies that underscore the increasingly important role of the pathologist in determining accurate diagnoses and facilitating appropriate treatment of patients with these disorders.
Serrated Polyps and Colorectal Cancer: New Pathway to Malignancy
Authors: Noffsinger Amy E.
Published: February 2009

Until recently, two major forms of colorectal epithelial polyp were recognized: the adenoma and the hyperplastic polyp. Adenomas were perceived to represent the precursor to colorectal cancer, whereas hyperplastic polyps were viewed as innocuous lesions with no potential for progression to malignancy. We now recognize, however, that the lesions formerly classified as hyperplastic actually represent a heterogeneous group of polyps, some of which have a significant risk for neoplastic transformation. These serrated polyps include not only hyperplastic polyps but also traditional serrated adenomas and sessile serrated adenomas. These polyps demonstrate characteristic molecular alterations not commonly seen in colorectal adenomas, and they probably progress to colorectal cancer by means of a new pathway: the serrated neoplasia pathway. The morphologic features of serrated colorectal lesions, the molecular alterations that characterize them, and their role in colorectal cancer development are discussed herein.
The Brainstem and Serotonin in the Sudden Infant Death Syndrome
Authors: Kinney Hannah C. , Richerson George B. , Dymecki Susan M. , Darnall Robert A. , Nattie Eugene E.
Published: February 2009

The sudden infant death syndrome (SIDS) is the sudden death of an infant under one year of age that is typically associated with sleep and that remains unexplained after a complete autopsy and death scene investigation. A leading hypothesis about its pathogenesis is that many cases result from defects in brainstem-mediated protective responses to homeostatic stressors occurring during sleep in a critical developmental period. Here we review the evidence for the brainstem hypothesis in SIDS with a focus upon abnormalities related to the neurotransmitter serotonin in the medulla oblongata, as these are the most robust pathologic findings to date. In this context, we synthesize the human autopsy data with genetic, whole-animal, and cellular data concerning the function and development of the medullary serotonergic system. These emerging data suggest an important underlying mechanism in SIDS that may help lead to identification of infants at risk and specific interventions to prevent death.
Influence of the Cavitation Model on the Simulation of Cloud Cavitation on 2D Foil Section
Authors: Frikha S. , Coutier-Delgosha O. , Astolfi J. A.
Published: February 2009

For numerical simulations of cavitating flows, many physical models are currently used. One approach is the void fraction transport equation‐based model including source terms for vaporization and condensation processes. Various source terms have been proposed by different researchers. However, they have been tested only in different flow configurations, which make direct comparisons between the results difficult. A comparative study, based on the expression of the source terms as a function of the pressure, is presented in the present paper. This analytical approach demonstrates a large resemblance between the models, and it also clarifies the influence of the model parameters on the vaporization and condensation terms and, therefore, on the cavity shape and behavior. Some of the models were also tested using a 2D CFD code in configurations of cavitation on two‐dimensional foil sections. Void fraction distributions and frequency of the cavity oscillations were compared to existing experimental measurements. These numerical results confirm the analytical study.
Some Estimates of Schrödinger‐Type Operators with Certain Nonnegative Potentials
Authors: Liu Yu , Ding Youzheng
Published: February 2009

We consider the Schrödinger‐type operator H = (−Δ)2 + V2, where the nonnegative potential V belongs to the reverse Hölder class for q1 ≥ n/2,  n ≥ 5. The Lp estimates of the operator ∇4H−1 related to H are obtained when and 1 < p ≤ q1/2. We also obtain the weak‐type estimates of the operator ∇4H−1 under the same condition of V.
Fabrication and Characterization of ZnO Nanowire Arrays with an Investigation into Electrochemical Sensing Capabilities
Authors: Weber Jessica , Jeedigunta Sathyaharish , Kumar Ashok
Published: February 2009

ZnO nanowire arrays were grown on a Si (100) substrate using the vapor‐liquid‐solid (VLS) method. ZnO nanowires were characterized by XRD, SEM, bright field TEM, and EDS. They were found to have a preferential orientation along the c‐axis. The as‐prepared sample was functionalized with glucose oxidase by physical adsorption. FTIR was taken before and after functionalization to verify the presence of the attached enzyme. Electrochemical measurements were performed on the nanowire array by differential pulse voltammetry in the range of −0.6 to 0.4 V. The nanoarray sensor displayed high sensitivity to glucose in the range of 1.0 ×10−4 to 1.0 ×10−2 mol L−1.
Series Solutions of Time‐Fractional PDEs by Homotopy Analysis Method
Authors: Abdulaziz O. , Hashim I. , Saif A.
Published: February 2009

The homotopy analysis method (HAM) is applied to solve linear and nonlinear fractional partial differential equations (fPDEs). The fractional derivatives are described by Caputo′s sense. Series solutions of the fPDEs are obtained. A convergence theorem for the series solution is also given. The test examples, which include a variable coefficient, inhomogeneous and hyperbolic‐type equations, demonstrate the capability of HAM for nonlinear fPDEs.
Stability of a Functional Equation Deriving from Cubic and Quartic Functions
Authors: Gordji M. Eshaghi , Ebadian A. , Zolfaghari S.
Published: February 2009

We obtain the general solution and the generalized
Ulam‐Hyers stability of the cubic and quartic functional equation
4(f(3x + y) + f(3x − y)) = −12(f(x + y) + f(x − y)) + 12(f(2x + y) + f(2x − y)) − 8f(y)−192f(x) + f(2y) + 30f(2x).
Burst-Mode Asynchronous Controllers on FPGA
Authors: Oliveira Duarte L. , Strum Marius , Sato Sandro S.
Published: February 2009

FPGAs have been mainly used to design synchronous circuits. Asynchronous design on FPGAs is difficult because the resulting circuit may suffer from hazard problems. We propose a method that implements a popular class of asynchronous circuits, known as burst mode, on FPGAs based on look-up table architectures. We present two conditions that, if satisfied, guarantee essential hazard-free implementation on any LUT-based FPGA. By doing that, besides all the intrinsic advantages of asynchronous over synchronous circuits, they also take advantage of the shorter design time and lower cost associated with FPGA designs.
Thermal Management with Solid‐Fluid Slip Irreversibility Treatment in Conjugate Microdevices
Authors: Ogedengbe E. O. B.
Published: February 2009

A numerical study of the effect of slip flow irreversibility and axial conduction in microdevices with a conjugate heat transfer between unmixed streams is presented. The effects of axial conduction due to parallel flows for thermal management in energy systems are investigated. Silicon substrate containing rectangular microchannels is simulated using a finite volume, staggered coupling of the pressure‐velocity fields. The entropy generation transport within the entire system is analyzed and coupled with the solution procedure. The effects of channel size perturbation, Reynolds number, and pressure ratios on the thermal performance and exergy destruction are presented. Comparative analysis of the axial conduction and flow irreversibility between parallel flow on thermal management is studied. A proton exchange membrane (PEM) fuel cell model is used as a quality indicator to access the importance of the exergy‐based design method.
Analytical Applications of Reactions of Iron(III) and Hexacyanoferrate(III) with 2,10‐Disubstituted Phenothiazines
Authors: Puzanowska-Tarasiewicz Helena , Karpińska Joanna , Kuźmicka Ludmiła
Published: February 2009

The presented review is devoted to analytical applications of reactions of Fe(III) and K3[Fe(CN)6] with 2,10‐disubstituted phenothiazines (PT). It was found that iron(III) and hexacyanoferrate(III) ions in acidic media easily oxidized PT with the formation of colored oxidation products. This property has been exploited for spectrophotometric determination of iron(III) ions and phenothiazines. Some flow‐injection procedures of the determination of PT based on the oxidation reaction by means of the above‐mentioned oxidants have been proposed. In the presented review, the application of 2,10‐disubstituted phenothiazines as indicators in complexometric titration of iron(III) as well as procedures of PT determination based on generation of ternary compound in the system Fe(III)‐SCN−‐ PT was also described.
γ-Cyclodextrin Increases Hydrolysis of Gangliosides by Sialidase fromArthrobacter ureafaciens: Hydrolysis of Gangliosides
Authors: Mitsumori Rie , Kato Tomohisa , Hatanaka Kenichi
Published: February 2009

Sialidase is a ubiquitous enzyme that catalyzes the hydrolytic removal of terminal sialic acid residues from oligosaccharides in glycolipids and glycoproteins. Ganglioside GM1 has been usually found to be resistant to various sialidases.Arthrobacter ureafacienssialidase has been reported to remove sialyl residues of ganglioside GM1 in the presence of bile salts. However, bile salts are difficult to be removed, and disturb HPTLC analysis. Usingγ-cyclodextrin (γ-CD) as a novel additive agent, ganglioside GM1 was efficiently hydrolyzed to asialo-GM1 byA. ureafacienssialidase.
A Novel Use for the Rigid Cystoscope: The Removal of Sacral Tacks after a Coloanal Anastamosis Dehiscence
Authors: Mahmalji W. , Mackenzie H. , Chopada A. , Raza A.
Published: February 2009

A 69-year-old female presented as an emergency with atrial fibrillation, which was treated with warfarin. She subsequently developed fresh rectal bleeding and after further investigations a Dukes B adenocarcinoma of the rectum was found. She subsequently underwent a low anterior resection, coloanal anastamosis and a defunctioning ileostomy. Three sterile surgical metallic tacks (pins) were inserted into the sacrum to stop brisk bleeding from the presacral venous plexus. Following discharge, she was readmitted with septic shock and a CT scan revealed a presacral fluid collection in the area surrounding the sacral tacks (pins) and an anastamotic dehiscence. The patient was not fit for further pelvic surgery to remove the tacks, so an alternative minimally invasive cystoscopic procedure was performed. The sacral tacks (pins) were removed by the urologist using a rigid cystoscope and cold cup biopsy forceps. To our knowledge, this is the first reported case in the literature.
The Efficiency of Operating Microscope Compared with Unaided Visual Examination, Conventional and Digital Intraoral Radiography for Proximal Caries Detection
Authors: Peker Ilkay , Toraman Alkurt Meryem , Bala Oya , Altunkaynak Bulent
Published: February 2009

Objective. The purpose of this study was to evaluate the efficiency of operating microscope compared with unaided visual examination, conventional and digital intraoral radiography for proximal caries detection.Materialsand Methods. The study was based on 48 extracted human posterior permanent teeth. The teeth were examined with unaided visual examination, operating microscope, conventional bitewing and digital intraoral radiographs. Then, true caries depth was determined by histological examination. The extent of the carious lesions was assessed by three examiners independently. One way variance of analysis (ANOVA) and Scheffe test were performed for comparison of observers, and the diagnostic accuracies of all systems were assessed from the area under the ROC curve ().Results. Statistically significant difference was found between observers (). There was a statistically significant difference between operating microscope-film radiography, operating microscope-RVG, unaided visual examination-film radiography, and unaided visual examination-RVG according to pairwise comparison ().Conclusion. The efficiency of operating microscope was found statistically equal with unaided visual examination and lower than radiographic systems for proximal caries detection.
A New Fork for Clinical Application: Targeting Forkhead Transcription Factors in Cancer
Authors: Yang Jer-Yen , Hung Mien-Chie
Published: February 2009

Forkhead O transcription factors (FOXO) play a pivotal role in the regulation of a myriad of cellular functions including cell cycle arrest, cell death, and protection from stress stimuli. Activation of cell survival pathways such as phosphoinositide-3-kinase/AKT/IKK or RAS/mitogen-activated protein kinase are known to phosphorylate FOXOs at different sites which cause FOXOs nuclear exclusion and degradation, resulting in the suppression of FOXO's transcriptional activity. Perturbation of FOXO's function leads to deregulated cell proliferation and accumulation of DNA damage, resulting in diseases such as cancer. Emerging evidence shows that active FOXO proteins are crucial for keeping cells in check; and inactivation of FOXO proteins is associated with tumorigenesis, including breast cancer, prostate cancer, glioblastoma, rhabdomyosarcoma, and leukemia. Moreover, clinically used drugs like paclitaxel, imatinib, and doxorubicin have been shown to achieve their therapeutic effects through activation of FOXO3a and FOXO3a targets. In this review, we will focus the novel functions of FOXOs revealed in recent studies and further highlight FOXOs as new therapeutic targets in a broad spectrum of cancers.
Prognostic Implications of and Relationship Between CpG Island Hypermethylation and Repetitive DNA Hypomethylation in Hepatocellular Carcinoma
Authors: Lee Hwan Seok , Kim Baek-Hee , Cho Nam-Yun , Yoo Eun Joo , Choi Minhee , Shin So-Hyun , Jang Ja-June , Suh Kyung-Suk , Kim Yong Sung , Kang Gyeong Hoon
Published: February 2009

Purpose: This study aims to determine the relationship between CpG island DNA hypermethylation and global genomic DNA hypomethylation and their prognostic implications in hepatocellular carcinoma. The association of DNA methylation changes with clinicopathologic factors and the chronological ordering of DNA methylation changes along multistep hepatocarcinogenesis were also assessed.

Experimental Design: Hepatocellular carcinoma (n = 20) and nonneoplastic liver samples (n = 72) were analyzed for their methylation status at 41 CpG island loci and 3 repetitive DNA elements (LINE-1, ALU, and SAT2) using MethyLight or combined bisulfite restriction analysis. After selection of 19 CpG island loci showing cancer-specific DNA methylation, another set of 99 hepatocellular carcinoma samples was analyzed for these loci.

Results: The number of methylated genes in hepatocellular carcinoma was significantly higher in hepatocellular carcinoma patients with a cirrhotic liver than in hepatocellular carcinoma patients with a noncirrhotic liver (9.9 versus 7.0, P = 0.001). Hepatocellular carcinoma from female patients showed a higher number of methylated genes than hepatocellular carcinoma from male patients (11.2 versus 8.4, P = 0.006). The genes CRABP1 and SYK showed significant association between CpG island hypermethylation and patients' poor survival. SAT2 hypomethylation occurred earlier than LINE-1 or ALU hypomethylation along the multistep hepatocarcinogenesis. Depending on the type of CpG island locus, a direct, inverse, or no relationship between CpG island hypermethylation and repetitive DNA hypomethylation was observed in hepatocellular carcinomas.

Conclusion: The varying relationships between the hypermethylation of individual CpG island loci and the hypomethylation of repetitive elements suggests that they are not mechanically linked. SYK and CRABP1 hypermethylation may serve as useful tumor markers for prognostication of hepatocellular carcinoma patients.
MDM2 Antagonist Nutlin-3 Displays Antiproliferative and Proapoptotic Activity in Mantle Cell Lymphoma
Authors: Tabe Yoko , Sebasigari Denise , Jin Linhua , Rudelius Martina , Davies-Hill Theresa , Miyake Kazunori , Miida Takashi , Pittaluga Stefania , Raffeld Mark
Published: February 2009

Purpose: Mantle cell lymphoma (MCL) has one of the poorest prognoses of the non-Hodgkin's lymphomas, and novel therapeutic approaches are needed. We wished to determine whether Nutlin-3, a novel small-molecule murine double minute 2 (MDM2) antagonist that efficiently activates TP53, might be effective in inducing cell death in MCL.

Experimental Design: MCL cell lines with known TP53 status were treated with Nutlin-3, and biological and biochemical consequences were studied. Synergies with the prototypic genotoxic agent doxorubicin and the novel proteasome inhibitor bortezomib were assessed.

Results: Nutlin-3 resulted in a reduction in cell proliferation/viability (IC50 < 10 μmol/L), an increase in the apoptotic fraction, and cell cycle arrest in wild-type (wt) TP53 Z-138 and Granta 519 cells. These effects were accompanied by TP53 accumulation and induction of TP53-dependent proteins p21, MDM2, Puma, and Noxa. Cell cycle arrest was characterized by suppression of S phase and an increase in the G0-G1 and G2-M fractions and accompanied by suppression of total and phosphorylated retinoblastoma protein and a decrease in G2-M-associated proteins cyclin B and CDC2. The combination of Nutlin-3 with doxorubicin or bortezomib was synergistic in wt-TP53 MCL cells. Nutlin-3 also induced cell cycle arrest and reduced cell viability in the mutant TP53 MINO cells but at a significantly higher IC50 (22.5 μmol/L). These effects were associated with induction of the TP53 homologue p73, slight increases in p21 and Noxa, and caspase activation. Nutlin-3 and bortezomib synergistically inhibited cell growth of MINO.

Conclusion: These findings suggest that the MDM2 antagonist Nutlin-3 may be an effective agent in the treatment of MCL with or without wt-TP53.
Functional B7.2 and B7-H2 Molecules on Myeloma Cells Are Associated with a Growth Advantage
Authors: Yamashita Taishi , Tamura Hideto , Satoh Chikako , Shinya Eiji , Takahashi Hidemi , Chen Lieping , Kondo Asaka , Tsuji Takashi , Dan Kazuo , Ogata Kiyoyuki
Published: February 2009

Purpose: B7 family molecules expressed on antigen-presenting cells stimulate or inhibit normal immune responses. The aim of this study was to investigate whether functional B7.2 and B7-H2 molecules are expressed on myeloma cells and, if so, whether they are associated with pathophysiology in myeloma.

Experimental Design: The expression of B7.2 and B7-H2 molecules on normal plasma and neoplastic (myeloma) plasma cells was analyzed. The cell proliferation and immunomodulatory function of myeloma cells related to B7.2 and B7-H2 expression were examined.

Results: Human myeloma cell lines commonly expressed B7.2 and B7-H2 molecules. B7.2 expression on plasma cells was more common in myeloma patients (n = 35) compared with that in patients with monoclonal gammopathy of unknown significance (n = 12) or hematologically normal individuals (n = 10). Plasma cells expressing B7-H2 were observed in myeloma patients alone, although rarely. Patients whose myeloma cells showed high B7.2 expression were more anemic and thrombocytopenic than other myeloma patients. The expression of these molecules was induced or augmented by cultivating myeloma cells with autologous stroma cells or tumor necrosis factor-α, a key cytokine in myeloma biology. Cell proliferation was more rapid in the B7.2+ and B7-H2+ populations compared with the B7.2− and B7-H2− populations, respectively, in the human myeloma cell lines examined. B7.2 and B7-H2 molecules on myeloma cells induced normal CD4+ T cells to proliferate and produce soluble factors, including interleukin-10 that stimulate myeloma cell proliferation.

Conclusions: Functional B7.2 and B7-H2 molecules detected on myeloma cells may be involved in the pathophysiology of myeloma.
Potential Therapeutic Effect of Glycogen Synthase Kinase 3β Inhibition against Human Glioblastoma
Authors: Miyashita Katsuyoshi , Kawakami Kazuyuki , Nakada Mitsutoshi , Mai Wei , Shakoori Abbas , Fujisawa Hironori , Hayashi Yutaka , Hamada Jun-ichiro , Minamoto Toshinari
Published: February 2009

Purpose: Glioblastoma represents the malignant brain tumor that is most refractory to treatment and in which the identification of molecular target(s) is urgently required. We investigated the expression, activity, and putative pathologic role of glycogen synthase kinase 3β (GSK3β), an emerging therapeutic target for neurodegenerative diseases, in human glioblastoma.

Experimental Design: The active fraction of GSK3β that is phosphorylated at the tyrosine 216 residue (pGSK3βY216) was identified in glioblastoma cell lines. GSK3β activity for phosphorylating its substrate was detected in these cells by nonradioisotopic in vitro kinase assay.

Results: Higher expression levels of GSK3β and pGSK3βY216 were frequently detected in glioblastomas compared with nonneoplastic brain tissues. Inhibition of GSK3β activity by escalating doses of a small-molecule inhibitor (AR-A014418) or inhibition of its expression by RNA interference induced the apoptosis and attenuated the survival and proliferation of glioblastoma cells in vitro. Inhibition of GSK3β was associated with increased expression of p53 and p21 in glioblastoma cells with wild-type p53 and with decreased Rb phosphorylation and expression of cyclin-dependent kinase 6 in all glioblastoma cell lines. Administration of AR-A014418 at a low dose significantly sensitized glioblastoma cells to temozolomide and 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea, chemotherapeutic agents used in the clinical setting, as well as to ionizing radiation.

Conclusion: These results indicate that GSK3β exerts a pathologic role by promoting the survival and proliferation of glioblastoma cells and by protecting them from apoptosis via the inactivation of p53- and/or Rb-mediated pathways. Consequently, we propose that GSK3β provides a potential therapeutic target in glioblastoma.
The Macrophage Colony-Stimulating Factor 1 Response Signature in Breast Carcinoma
Authors: Beck Andrew H. , Espinosa Inigo , Edris Badreddin , Li Rui , Montgomery Kelli , Zhu Shirley , Varma Sushama , Marinelli Robert J. , van de Rijn Matt , West Robert B.
Published: February 2009

Purpose: Macrophages play an important role in breast carcinogenesis. The pathways that mediate the macrophage contribution to breast cancer and the heterogeneity that exists within macrophages are incompletely understood. Macrophage colony-stimulating factor 1 (CSF1) is the primary regulator of tissue macrophages. The purpose of this study was to define a novel CSF1 response signature and to evaluate its clinical and biological significance in breast cancer.

Experimental Design: We defined the CSF1 response signature by identifying genes overexpressed in tenosynovial giant cell tumor and pigmented villonodular synovitis (tumors composed predominantly of macrophages recruited in response to the overexpression of CSF1) compared with desmoid-type fibromatosis and solitary fibrous tumor. To characterize the CSF1 response signature in breast cancer, we analyzed the expression of CSF1 response signature genes in eight published breast cancer gene expression data sets (n = 982) and did immunohistochemistry and in situ hybridization for CSF1 response genes on a breast cancer tissue microarray (n = 283).

Results: In both the gene microarray and tissue microarray analyses, a consistent subset (17-25%) of breast cancers shows the CSF1 response signature. The signature is associated with higher tumor grade, decreased expression of estrogen receptor, decreased expression of progesterone receptor, and increased TP53 mutations (P < 0.001).

Conclusions: Our data show that the CSF1 response signature is consistently seen in a subset of breast carcinomas and correlates with biological features of the tumor. Our findings provide insight into macrophage biology and may facilitate the development of personalized therapy for patients most likely to benefit from CSF1-targeted treatments.
Identification and Characterization of a Novel Melanoma Tumor Suppressor Gene on Human Chromosome 6q21
Authors: Fung Jackie Mei-Wah , Smith Ross , Brown Melissa A. , Lau Sze Hang , Xie Dan , Lau George K. , Guan Xin-Yuan
Published: February 2009

Purpose: By characterizing a complex chromosome rearrangement involving 6q and 17p in melanoma cell line UACC-930, we isolated a candidate tumor suppressor gene at 6q21, named prenyl diphosphate synthase subunit 2 (PDSS2), which was interrupted by an inversion breakpoint. The purpose of this study was to determine the tumor-suppressive potential of PDSS2 in the development of melanoma.

Experimental Design: To isolate the rearranged 6q in UACC-930 cells, a bacterial artificial chromosome clone (RP1-67A8) covering the breakpoint at 6q21 was digested with HindIII and each DNA fragment was used as the probe for the breakpoint in Southern blotting. The HindIII fragment probe covering the breakpoint was then used to screen an EcoRI-digested DNA library generated from UACC-930. To characterize the tumor-suppressive potential of PDSS2, PDSS2 was stably transfected into a highly tumorigenic melanoma cell line, UACC-903. The tumor-suppressive function of PDSS2 was shown by both in vitro and in vivo assays. The differential expression of PDSS2 in benign nevi and primary melanoma samples was also studied.

Results: Down-regulation of PDSS2 was observed in 59 of 87 (67.8%) primary melanomas, which was significantly higher than that in benign nevi (7 of 66, 10.6%; P < 0.001). In addition, an overexpression of the PDSS2 in UACC-903 cells could inhibit tumor cell growth, decrease the colony-forming ability in soft agar, and totally abrogate the tumorigenicity of UACC-903 in nude mice.

Conclusions: Our results support the proposal that PDSS2 is a novel tumor suppressor gene that plays an important role in the development of malignant melanoma.
Immunomodulation of FOXP3+ Regulatory T Cells by the Aromatase Inhibitor Letrozole in Breast Cancer Patients
Authors: Generali Daniele , Bates Gaynor , Berruti Alfredo , Brizzi Maria P. , Campo Leticia , Bonardi Simone , Bersiga Alessandra , Allevi Giovanni , Milani Manuela , Aguggini Sergio , Dogliotti Luigi , Banham Alison H. , Harris Adrian L. , Bottini Alberto , Fox Stephen B.
Published: February 2009

Purpose: We have shown previously that tumor infiltration by FOXP3+ regulatory T cells (Treg) is associated with increased relapse and shorter survival of patients with both in situ and invasive breast cancer. Because estrogen regulates Treg numbers in mice and promotes the proliferation of human Tregs, we hypothesized that blocking estrogen receptor-α signaling would abrogate Tregs and be associated with response to hormonal therapy and increased survival.

Experimental Design: FOXP3+ Tregs were quantified in tumor samples collected at baseline by incisional biopsy and after 6 months at definitive surgery in 83 elderly breast cancer patients (T2-4 N0-1) enrolled in a randomized phase II trial based on 6 months of primary letrozole (2.5 mg/d) or 6 months of letrozole plus oral “metronomic” cyclophosphamide (50 mg/d).

Results: Treg number ranged from 0 to 380 (median, 30) before treatment and from 0 to 300 (median, 8) after treatment. There was a significant reduction in Tregs in letrozole and letrozole-cyclophosphamide patients (P < 0.0001 and P < 0.002, respectively) after treatment. Treg number at residual histology was inversely related with response (P < 0.03 and P = 0.50, respectively) and a greater Treg reduction was observed in responding patients (P < 0.03).

Conclusion: This study suggests that aromatase inhibitors may have an indirect antitumor mechanism of action through reducing Tregs in breast tumors and may be of use in estrogen receptor-α-negative tumors in combination with immunotherapy approaches.
B7 Blockade Alters the Balance between Regulatory T Cells and Tumor-reactive T Cells for Immunotherapy of Cancer
Authors: Zhou Penghui , Zheng Xincheng , Zhang Huiming , Liu Yang , Zheng Pan
Published: February 2009

Purpose: In prostate cancer-bearing host, regulatory T (Treg) cells restrain activity of tumor antigen-specific T cells. Because B7:CD28 interactions are needed for both function of CD4+CD25+ Treg cells and CD8+ effective T cells, targeting this pathway may help to overcome the immunotherapy barriers.

Experimental Design: The anti-B7-1/B7-2 monoclonal antibodies were administered to a transgenic mouse model of prostate cancer (TRAMP) ectopically expressing SV40 large T antigen in different tumor development stages for prevention and therapy of prostate cancer. The treatment was also tested in treating transplanted MC38 colon adenocarcinoma in mice.

Results: Here, we showed that short-term administration of anti-B7-1/B7-2 monoclonal antibodies in TRAMP mice leads to significant inhibited primary tumor growth and the size of metastatic lesions. The treatment is effective to inhibit MC38 colon cancer growth. Correspondingly, this treatment results in a transient reduction of Treg in both thymus and the periphery. In vivo cytotoxicity assay revealed T antigen-specific CTL effectors in anti-B7-treated but not control IgG-treated TRAMP mice.

Conclusions: Transient blockade of B7-1/B7-2 alters the balance between Treg and cancer-reactive T cells to enhance cancer immunotherapy.
Grape Seed Proanthocyanidins Inhibit the Growth of Human Non-Small Cell Lung Cancer Xenografts by Targeting Insulin-Like Growth Factor Binding Protein-3, Tumor Cell Proliferation, and Angiogenic Factors
Authors: Akhtar Suhail , Meeran Syed M. , Katiyar Nandan , Katiyar Santosh K.
Published: February 2009

Purpose: Lung cancer is a leading cause of cancer-related deaths worldwide. Here, we assessed the chemotherapeutic effect of grape seed proanthocyanidins (GSPs) on human non-small cell lung cancer (NSCLC) cells in vitro and in vivo using a tumor xenograft model.

Experimental Design: The effects of GSPs on human NSCLC cell lines in terms of cellular proliferation were determined. The chemotherapeutic effects of a GSP- supplemented AIN76A control diet fed to nude mice bearing tumor xenografts (A549 and H1299) were evaluated in terms of biomarkers of cell proliferation and angiogenesis and on insulin-like growth factor binding protein-3 using immunohistochemical detection, ELISA, and Western blotting.

Results: In vitro treatment of NSCLC cells with GSPs resulted in inhibition of cellular proliferation. Administration of GSPs (0.1%, 0.2%, and 0.5%, w/w) as a supplement of an AIN76A control diet resulted in a dose-dependent inhibition of the growth of NSCLC (A549 and H1299) tumor xenografts in athymic nude mice (25-76%; P < 0.05-0.001). The growth-inhibitory effect of GSPs on the NSCLC xenograft tumors was associated with the enhancement of the levels of insulin-like growth factor binding protein-3 in the tumor microenvironment and plasma and antiproliferative, antiangiogenic, and proapoptotic effects.

Conclusions: This preclinical study reveals for the first time that dietary GSPs have the ability to inhibit the growth of human NSCLC tumor xenografts grown in vivo in athymic nude mice. More studies are needed to develop GSPs as a pharmacologically safe agent for the prevention of lung cancer in humans.
An Interstitial Deletion at 3p21.3 Results in the Genetic Fusion ofMLH1andITGA9in a Lynch Syndrome Family
Authors: Meyer Claus , Brieger Angela , Plotz Guido , Weber Nicole , Passmann Sandra , Dingermann Theo , Zeuzem Stefan , Trojan Joerg , Marschalek Rolf
Published: February 2009

Purpose: Germline mutations in DNA mismatch repair genes, mainly MLH1 or MSH2, have been shown to predispose with high penetrance for the development of the clinical phenotype of hereditary nonpolyposis colorectal cancer (Lynch syndrome). Here, we describe the discovery and first functional characterization of a novel germline MLH1 mutant allele.

Experimental Design: A large kindred including 54 potential carriers was investigated at the molecular level by using different types of PCR experiments, gene cloning, transfection studies, Western blot experiments, and mismatch repair assays to identify and characterize a novel MLH1 mutant allele. Twenty-two of 54 putative carriers developed colon cancer or other tumors, including breast cancer.

Results: The identified MLH1 mutant allele emerged from an interstitial deletion on chromosome 3p21.3, leading to an in-frame fusion of MLH1 (exons 1-11) with ITGA9 (integrin α 9; exons 17-28). The deleted area has a size of about 400 kb; codes for LRRFIP2 (leucine-rich repeat in flightless interaction protein 2), GOLGA4 (Golgi autoantigen, golgin subfamily a, 4), and C3orf35/APRG1 (chromosome 3 open reading frame 35/AP20 region protein 1); and partly disrupts the AP20 region implicated in major epithelial malignancies. Tumor cells lost their second MLH1 allele. The MLH1•ITGA9 fusion protein provides no capability for DNA mismatch repair. Murine fibroblasts, expressing a doxycycline-inducible MLH1•ITGA9 fusion gene, exhibit a loss–of–contact inhibition phenotype.

Conclusions: This is the first description of a functional gene fusion of the human MLH1 gene, resulting in the loss of mismatch repair capabilities. The MLH1•ITGA9 fusion allele, together with deletions of the AP20 region, presumably defines a novel subclass of Lynch syndrome patients, which results in an extended tumor spectrum known from hereditary nonpolyposis colorectal cancer and Muir-Torre syndrome patients.
Dual Degradation of Aurora A and B Kinases by the Histone Deacetylase Inhibitor LBH589 Induces G2-M Arrest and Apoptosis of Renal Cancer Cells
Authors: Cha Tai-Lung , Chuang Mei-Jen , Wu Sheng-Tang , Sun Guang-Huan , Chang Sun-Yran , Yu Dah-Shyong , Huang Shih-Ming , Huan Steven Kuan-Hua , Cheng Tse-Chou , Chen Tzu-Ting , Fan Pao-Luo , Hsiao Pei-Wen
Published: February 2009

Purpose: This study is aimed at investigating antineoplastic efficacy of histone deacetylase inhibitor (HDACI) LBH589 on renal cell carcinoma (RCC) and elucidating the novel molecular mechanisms involved in growth arrest and apoptosis by targeting the important nonhistone molecules.

Experimental Design: We analyzed the growth-inhibitory effect of LBH589 on RCC by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in vitro and antitumor efficacy by xenograft experiments in vivo. To verify the associated molecular mechanisms involved in LBH589-mediated cell death and cell cycle progression by Western blotting and fluorescence-activated cell sorting analysis.

Results: HDACI LBH589 induced degradation of both Aurora A and B kinases through a proteasome-mediated pathway by targeting HDAC3 and HDAC6. The dual degradation of Aurora A and B kinases mediated by LBH589 resulted in inducing G2-M arrest and apoptosis of renal cancer cell lines and our results also showed that LBH589 potently inhibited renal cancer cell growth in vitro and suppressed tumor formation in vivo. The Aurora A and B kinases and HDAC3 are overexpressed in the human RCC tumor tissues examined, which make them perfect targets for HDACI LBH589 treatment.

Conclusions: Our in vitro and in vivo data showed that LBH589 has potent anticancer effect of renal cancer cells. LBH589 and other HDACI treatment resulted in inducing G2-M arrest and apoptosis of renal cancer cells through degradation of Aurora A and B kinases by inhibition of HDAC3 and HDAC6. The clinical efficacy of LBH589 in the treatment of patients with metastatic RCC, especially those with high Aurora kinase and HDAC expression, is worthy of further investigation.
Differential Diagnosis of Pulmonary Carcinoma Following Head and Neck Cancer by Genetic Analysis
Authors: Geurts T.W. , van Velthuysen M.L.F , Broekman F. , Hooft van Huysduynen T. , van den Brekel M.W.M. , van Zandwijk N. , van Tinteren H. , Nederlof P. , Balm A.J.M. , Brakenhoff R.H.
Published: February 2009

Purpose: Patients with head and neck cancer often develop a lung tumor that can be diagnosed as distant metastasis (DM) or second primary tumor (SPT). In this study, we use TP53 mutation analysis for validation of an allelic loss marker panel and a decision algorithm for distinguishing between DM and SPT.

Experimental Design: Tumor pairs of 39 patients were analyzed for TP53 mutations, for patterns of allelic loss and immunohistochemical analysis of p53 expression. Results of these three analyses were compared, using mutation analysis as gold standard.

Results: Loss of heterozygosity (LOH) analysis indicated DM in 15 and SPT in 23 cases (one inconclusive). TP53 mutation analysis was informative in 15 cases. Based on the p53 mutation status alone, nine tumors were diagnosed as SPT and six as DM. In all 15 cases the LOH analysis was in concordance with the TP53 mutation analysis. Immunostaining for p53 showed promise as a first scan to diagnose lung tumors as SPT but cannot be used to diagnose DM.

Conclusion: The TP53 mutation data validate the suitability of the LOH marker panel and decision algorithm for differential diagnosis of DM and SPT in the lung. LOH analysis can theoretically be exploited in almost all cases and is less laborious than TP53 mutation analysis.
Response to Sunitinib Malate in Advanced Alveolar Soft Part Sarcoma
Authors: Stacchiotti Silvia , Tamborini Elena , Marrari Andrea , Brich Silvia , Rota Sara Arisi , Orsenigo Marta , Crippa Flavio , Morosi Carlo , Gronchi Alessandro , Pierotti Marco A. , Casali Paolo G. , Pilotti Silvana
Published: February 2009

Purpose: Alveolar soft part sarcoma (ASPS) is a rare, chemoresistant soft tissue sarcoma. ASPS harbors the t(17-X) (p11.2;q25) translocation, resulting in the ASPACR1-TFE3 fusion protein, causing MET autophosphorylation and activation of downstream signaling. The tumor vascular pattern prompted us to use sunitinib malate (SM), a tyrosine kinase inhibitor with antiangiogenic properties.

Experimental Design: Since July 2007, five patients with progressive metastatic ASPS have been treated with continuous SM 37.5 mg/d on a named basis. Four patients are evaluable for response. In four cases, cryopreserved material was available. Upstream and downstream targets of receptor tyrosine kinase (RTK) pathways, as well as mechanisms of activation, were investigated by biochemical profiles, including human phospho-receptor RTK antibody arrays and immunoprecipitation/Western blotting, molecular analyses, immunohistochemistry, and fluorescence in situ hybridization analyses.

Results: After 3 months, two patients had RECIST (response evaluation criteria in solid tumor) partial response, as well as positron emission tomography response and subjective improvement. One had a RECIST stable disease. One progressed and stopped treatment. One patient is still responding after 12 months. The upstream analysis showed activation of all the platelet-derived growth factor receptor (PDGFR) family members, as well as epidermal growth factor receptor, MET families, and RET. Vascular endothelial growth factor receptors (VEGFR1 and VEGFR2) were activated only in one case. The downstream target analysis showed strong activation of phosphatidylinositol 3-kinase/AKT, extracellular signal-regulated kinase 1/2, and mTOR and its targets (S6K and S6). The absence of any upstream mTOR effector deregulation and the presence of RTK cognate ligands support an autocrine-paracrine activation loop mechanism.

Conclusion: SM may have antitumor activity in ASPS, possibly through a mechanism involving PDGFR and RET. The role of MET, epidermal growth factor receptor, and mTOR, as well as PDGFR inhibition, needs to be further explored.
Phase II Trial of Gliadel plus O6-Benzylguanine in Adults with Recurrent Glioblastoma Multiforme
Authors: Quinn Jennifer A. , Jiang Sara Xiaoyin , Carter James , Reardon David A. , Desjardins Annick , Vredenburgh James J. , Rich Jeremy N. , Gururangan Sridharan , Friedman Allan H. , Bigner Darell D. , Sampson John H. , McLendon Roger E. , Herndon James E. , Threatt Stevie , Friedman Henry S.
Published: February 2009

Purpose: This phase II trial was designed to define the efficacy of Gliadel wafers in combination with an infusion of O6-benzylguanine (O6-BG) that suppresses tumor O6-alkylguanine-DNA alkyltransferase (AGT) levels in patients with recurrent glioblastoma multiforme for 5 days and to evaluate the safety of this combination therapy.

Experimental Design: This was a phase II, open-label, single center trial. On gross total resection of the tumor, up to eight Gliadel wafers were implanted. Bolus infusion of O6-BG was administered at 120 mg/m2 over 1 hour on days 1, 3, and 5, along with a continuous infusion at 30 mg/m2/d. The primary end points were 6-month overall survival (OS) and safety, and the secondary end points were 1-year, 2-year, and median OS.

Results: Fifty-two patients were accrued. The 6-month OS was 82% [95% confidence interval (95% CI), 72-93%]. The 1- and 2-year OS rates were 47% (95% CI, 35-63%) and 10% (95% CI, 3-32%), respectively. The median OS was 50.3 weeks (95% CI, 36.1-69.4 weeks). Treatment-related toxicity with this drug combination included grade 3 hydrocephalus (9.6%), grade 3 cerebrospinal fluid (CSF) leak (19.2%), and grade 3 CSF/brain infection (13.4%).

Conclusion: The efficacy of implanted Gliadel wafers may be improved with the addition of O6-BG. Although systemically administered O6-BG can be coadministered with Gliadel wafers safely, it may increase the risk of hydrocephalus, CSF leak, and CSF/brain infection. Future trials are required to verify that inhibition of tumor AGT levels by O6-BG results in increased efficacy of Gliadel wafers without added toxicity.
Expression of a BTB/POZ Protein, NAC1, Is Essential for the Proliferation of Normal Cyclic Endometrial Glandular Cells and Is Up-regulated by Estrogen
Authors: Ishibashi Masako , Nakayama Kentaro , Yeasmin Shamima , Katagiri Atsuko , Iida Kouji , Nakayama Naomi , Miyazaki Kohji
Published: February 2009

Purpose: The purpose of this study was to investigate the expression and localization of NAC1, a member of the BTB/POZ gene family in the human cyclic endometrium.

Experimental Design: NAC1 expression in normal cyclic endometrium was assessed by immunohistochemistry, and data on clinical variables were collected by retrospective chart review. To elucidate the molecular mechanisms of NAC1 expression in the normal endometrium endometrial carcinoma cell lines (Ishikawa, HHUA; ER+, PR+) and primary cultured normal endometria were tested in a sex steroid induction assay and a NAC1 knockdown assay using siRNA.

Results: Expression of NAC1 in glandular cells was significantly higher in the early and mid proliferative phases than in the other menstrual phases. Both NAC1 RNA and protein expression were up-regulated by treatment with 10 nmol/L 17β-Estradiol (E2) in Ishikawa, HHUA and primary cultured normal endometrial cells. The estrogen receptor antagonist ICI 182,780 significantly attenuated E2-induced NAC1 expression. NAC1 gene knockdown inhibited cell growth and induced apoptosis in Ishikawa, HHUA, and normal endometria, all of which expressed NAC1. Furthermore, NAC1 siRNA significantly abrogated estrogen-driven cellular proliferation in Ishikawa, HHUA, and primary cultured normal endometrial cells, whereas the control siRNA had no effect on cell growth in any of these cells.

Conclusions: These findings suggest that NAC1 is functionally involved in E2-induced cell growth of the normal endometrial glandular cells. Because NAC1 is thought to have oncogenic potential, the current findings may provide new insight into the mechanism of estrogen induced endometrial carcinogenesis.
Induction of p53-Specific Immunity by a p53 Synthetic Long Peptide Vaccine in Patients Treated for Metastatic Colorectal Cancer
Authors: Speetjens Frank M. , Kuppen Peter J.K. , Welters Marij J.P. , Essahsah Farah , Voet van den Brink Anne Marie E.G. , Lantrua M. Graziella Kallenberg , Valentijn A. Rob P.M. , Oostendorp Jaap , Fathers Lorraine M. , Nijman Hans W. , Drijfhout Jan W. , van de Velde Cornelis J.H. , Melief Cornelis J.M. , van der Burg Sjoerd H.
Published: February 2009

Purpose: The tumor-associated self-antigen p53 is commonly overexpressed in cancer, including colorectal cancer, and can serve as a target for immunotherapy. The safety and immunogenicity of a p53 synthetic long peptide (p53-SLP) vaccine were investigated in patients treated for metastatic colorectal cancer.

Experimental Design: Ten patients were vaccinated twice with a set of 10 overlapping p53-SLP in a phase I/II trial. Both the safety and the breadth, magnitude, and polarization of vaccine-induced p53-specific T cells was evaluated in blood samples drawn before and after vaccination by IFN-γ enzyme-linked immunospot, proliferation, cytokine secretion, and multiparameter flow cytometry. The migratory capacity of p53-specific T cells was evaluated by assessing their presence in a biopsy of the second vaccination site.

Results: Toxicity was limited to grade 1/2, mostly at the vaccination site. p53-specific T-cell responses were induced in 9 of 10 colorectal cancer patients as measured by IFN-γ enzyme-linked immunospot, proliferation, and cytokine bead array. In 6 of 9 tested patients, p53-specific T-cell reactivity persisted at least 6 months. Furthermore, p53-specific T cells isolated from the vaccination site were characterized as CD4+ T cells producing both T-helper types 1 and 2 cytokines on stimulation with p53 peptide and p53 protein. Multiparameter flow cytometry revealed that only a minor population of the p53-specific CD4+ T cells was optimally polarized.

Conclusions: The p53-SLP vaccine is safe and capable to induce p53-specific T-cell responses in patients treated for colorectal cancer. New trials should focus on improving the polarization of the p53-SLP vaccine-induced T-cell response.