bluuebunny/crossref_metadata_2025 huggingface dataset:
Microarray Detection Call Methodology as a Means to Identify and Compare Transcripts Expressed within Syncytial Cells from Soybean (Glycine max) Roots Undergoing Resistant and Susceptible Reactions to the Soybean Cyst Nematode (Heterodera glycines)
Authors: Klink Vincent P. , Overall Christopher C. , Alkharouf Nadim W. , MacDonald Margaret H. , Matthews Benjamin F.
Published: May 2010

Background. A comparative microarray investigation was done using detection call methodology (DCM) and differential expression analyses. The goal was to identify genes found in specific cell populations that were eliminated by differential expression analysis due to the nature of differential expression methods. Laser capture microdissection (LCM) was used to isolate nearly homogeneous populations of plant root cells.Results. The analyses identified the presence of 13,291 transcripts between the 4 different sample types. The transcripts filtered down into a total of 6,267 that were detected as being present in one or more sample types. A comparative analysis of DCM and differential expression methods showed a group of genes that were not differentially expressed, but were expressed at detectable amounts within specific cell types.Conclusion. The DCM has identified patterns of gene expression not shown by differential expression analyses. DCM has identified genes that are possibly cell-type specific and/or involved in important aspects of plant nematode interactions during the resistance response, revealing the uniqueness of a particular cell population at a particular point during its differentiation process.
Nonintegrating Lentiviral Vector-Based Vaccine Efficiently Induces Functional and Persistent CD8+ T Cell Responses in Mice
Authors: Negri Donatella R. M. , Michelini Zuleika , Baroncelli Silvia , Spada Massimo , Vendetti Silvia , Bona Roberta , Leone Pasqualina , Klotman Mary E. , Cara Andrea
Published: May 2010

CD8+ T cells are an essential component of an effective host immune response to tumors and viral infections. Genetic immunization is particularly suitable for inducing CTL responses, because the encoded proteins enter the MHC class I processing pathway through either transgene expression or cross-presentation. In order to compare the efficiency and persistence of immune response induced by genetic vaccines, BALB/c mice were immunized either twice intramuscularly with DNA plasmid expressing a codon-optimized HIV-1 gp120 Envelope sequence together with murine GM-CSF sequence or with a single immunization using an integrase defective lentiviral vector (IDLV) expressing the same proteins. Results strongly indicated that the schedule based on IDLV vaccine was more efficient in inducing specific immune response, as evaluated three months after the last immunization by IFN ELISPOT in both splenocytes and bone marrow- (BM-) derived cells, chromium release assay in splenocytes, and antibody detection in sera. In addition, IDLV immunization induced high frequency of polyfunctional CD8+ T cells able to simultaneously produce IFN, TNF and IL2.
Regulation of the Induction and Function of Cytotoxic T Lymphocytes by Natural Killer T Cell
Authors: Ito Hiroyasu , Seishima Mitsuru
Published: May 2010

Cytotoxic T lymphocytes (CTLs) play a crucial role in the infections and the antitumor immunity. Induction and activation of antigen-specific CTLs is an important strategy in immunotherapy for various diseases, and several researchers have focused on the modulation of CTL induction and function. Natural killer T (NKT) cells are an important focus area of researchers studying immunomodulatory responses to tumors and infectious diseases. CD1d-restricted NKT cells consist of type I NKT cells and type II NKT cells. -galactosylceramide (-GalCer)-activated type I NKT cells secrete both Th1 (e.g., IFN-) and Th2 cytokines, affect the expression of costimulatory molecules in immune cells, and regulate the host immune system. Type II NKT cells, however, are stimulated by sulfatide, a self-glycolipid derived from myelin, and play an immunosuppressive role in animal model of autoimmune diseases. CTL generation, activation, and suppression are strongly affected by activated type I and type II NKT cells. Thus, the regulation of these NKT cells leads to the modification of CTL function. CTLs contribute to antimicrobial responses, antitumor immune and autoimmune responses. Understanding the role of NKT cells in the regulation of CTL generation, activation, and suppression enable the development of novel treatment strategies for these diseases.
Analysis of Genetic Diversity in Astragalus rhizanthus Benth. ssp. rhizanthus var. rhizanthus (Fabaceae) Using Molecular Markers from India
Authors: Anand Kumar Kamal , Srivastava Ravi Kumar , Chaudhary Lal Babu
Published: May 2010

Astragalus rhizanthus with three infraspecific taxa (i.e., A. rhizanthus ssp. rhizanthus var. rhizanthus, A. rhizanthus ssp. rhizanthus var. pindreensis, and A. rhizanthus ssp. candolleanus) is widely scattered in the Himalaya from Jammu and Kashmir to Uttarakhand provinces in India. Among them, A. rhizanthus ssp. rhizanthus var. rhizanthus exhibits enormous morphological diversity throughout its range of distribution. An assessment of genetic diversity studies was undertaken to understand the level and pattern of diversity, using Inter simple sequence repeats (ISSR), Random amplified polymorphic DNA (RAPD) and Directed amplification of minisatellite DNA (DAMD) profiles. Fifteen ISSR, 18 RAPD and 6 DAMD primers were used to unravel the diversity among 20 genotypes collected from the known localities in the Indian Himalaya. A total of 242 bands from ISSR, 352 from RAPD and 142 from DAMD were obtained with an average of 92.23% polymorphism in the species. Pair-wise genetic similarity for the cumulative data was determined using Jaccard's similarity coefficient which varied from 0.19 to 0.84. A combined UPGMA dendrogram was generated which revealed that different genotypes exhibited their affinity according to their geographical distribution. Tree topology suggests the existence of two distinct groups of the genotypes. Gene diversity (H) and Shannon's information index (I) were estimated and these values were found higher in the genotypes collected from Jammu and Kashmir than Himachal Pradesh.
Water and Wastewater Pipe Nondestructive Evaluation and Health Monitoring: A Review
Authors: Rizzo Piervincenzo
Published: May 2010

Civil infrastructures such as bridges, buildings, and pipelines ensure society's economic and industrial prosperity. Specifically, pipe networks assure the transportation of primary commodities such as water, oil, and natural gas. The quantitative and early detection of defects in pipes is critical in order to avoid severe consequences. As a result of high-profile accidents and economic downturn, research and development in the area of pipeline inspection has focused mainly on gas and oil pipelines. Due to the low cost of water, the development of nondestructive inspection (NDI) and structural health monitoring (SHM) technologies for fresh water mains and sewers has received the least attention. Moreover, the technical challenges associated with the practical deployment of monitoring system demand synergistic interaction across several disciplines, which may limit the transition from laboratory to real structures. This paper presents an overview of the most used NDI/SHM technologies for freshwater pipes and sewers. The challenges that said infrastructures pose with respect to oil and natural gas pipeline networks will be discussed. Finally, the methodologies that can be translated into SHM approaches are highlighted.
-Thalassemia: HiJAKing Ineffective Erythropoiesis and Iron Overload
Authors: Melchiori Luca , Gardenghi Sara , Rivella Stefano
Published: May 2010

-thalassemia encompasses a group of monogenic diseases that have in common defective synthesis of -globin. The defects involved are extremely heterogeneous and give rise to a large phenotypic spectrum, with patients that are almost asymptomatic to cases in which regular blood transfusions are required to sustain life. As a result of the inefficient synthesis of -globin, the patients suffer from chronic anemia due to a process called ineffective erythropoiesis (IE). The sequelae of IE lead to extramedullary hematopoiesis (EMH) with massive splenomegaly and dramatic iron overload, which in turn is responsible for many of the secondary pathologies observed in thalassemic patients. The processes are intimately linked such that an ideal therapeutic approach should address all of the complications. Although -thalassemia is one of the first monogenic diseases to be described and represents a global health problem, only recently has the scientific community started to focus on the real molecular mechanisms that underlie this disease, opening new and exciting therapeutic perspectives for thalassemic patients worldwide.
Inhibiting the Cyclin-Dependent Kinase CDK5 Blocks Pancreatic Cancer Formation and Progression through the Suppression of Ras-Ral Signaling
Authors: Feldmann Georg , Mishra Anjali , Hong Seung-Mo , Bisht Savita , Strock Christopher J. , Ball Douglas W. , Goggins Michael , Maitra Anirban , Nelkin Barry D.
Published: May 2010

Cyclin-dependent kinase 5 (CDK5), a neuronal kinase that functions in migration, has been found to be activated in some human cancers in which it has been implicated in promoting metastasis. In this study, we investigated the role of CDK5 in pancreatic cancers in which metastatic disease is most common at diagnosis. CDK5 was widely active in pancreatic cancer cells. Functional ablation significantly inhibited invasion, migration, and anchorage-independent growth in vitro, and orthotopic tumor formation and systemic metastases in vivo. CDK5 blockade resulted in the profound inhibition of Ras signaling through its critical effectors RalA and RalB. Conversely, restoring Ral function rescued the effects of CDK5 inhibition in pancreatic cancer cells. Our findings identify CDK5 as a pharmacologically tractable target to degrade Ras signaling in pancreatic cancer. Cancer Res; 70(11); 4460–9. ©2010 AACR.
Cadherin-11 Increases Migration and Invasion of Prostate Cancer Cells and Enhances their Interaction with Osteoblasts
Authors: Huang Chih-Fen , Lira Cristina , Chu Khoi , Bilen Mehmet Asim , Lee Yu-Chen , Ye Xiangcang , Kim Soo Mi , Ortiz Angelica , Wu Fe-Lin Lin , Logothetis Christopher J. , Yu-Lee Li-Yuan , Lin Sue-Hwa
Published: May 2010

Cell adhesion molecules have been implicated in the colonization of cancer cells to distant organs. Prostate cancer (PCa) has a propensity to metastasize to bone, and cadherin-11, which is an osteoblast cadherin aberrantly expressed in PCa cells derived from bone metastases, has been shown to play a role in the metastasis of PCa cells to bone. However, the mechanism by which cadherin-11 is involved in this process is not known. Here, we show that expression of cadherin-11 in cadherin-11–negative C4-2B4 cells increases their spreading and intercalation into an osteoblast layer and also stimulates C4-2B4 cell migration and invasiveness. The downregulation of cadherin-11 in cadherin-11–expressing metastatic PC3 cells decreases cell motility and invasiveness. Further, both the juxtamembrane (JMD) and β-catenin binding domains (CBS) in the cytoplasmic tail of cadherin-11 are required for cell migration and invasion, but not spreading. Gene array analyses showed that several invasion-related genes, including MMP-7 and MMP-15, are upregulated in cadherin-11–expressing, but not in cad11-ΔJMD–expressing or cad11-ΔCBS–expressing, C4-2B4 cells. These observations suggest that cadherin-11 not only provides a physical link between PCa cells and osteoblasts but also increases PCa cell motility and invasiveness that may facilitate the metastatic colonization of PCa cells in bone. Cancer Res; 70(11); 4580–9. ©2010 AACR.
Preventing the Activation or Cycling of the Rap1 GTPase Alters Adhesion and Cytoskeletal Dynamics and Blocks Metastatic Melanoma Cell Extravasation into the Lungs
Authors: Freeman Spencer A. , McLeod Sarah J. , Dukowski Janet , Austin Pamela , Lee Crystal C.Y. , Millen-Martin Brandie , Kubes Paul , McCafferty Donna-Marie , Gold Michael R. , Roskelley Calvin D.
Published: May 2010

The Rap1 GTPase is a master regulator of cell adhesion, polarity, and migration. We show that both blocking Rap1 activation and expressing a constitutively active form of Rap1 reduced the ability of B16F1 melanoma cells to extravasate from the microvasculature and form metastatic lesions in the lungs. This correlated with a decreased ability of the tumor cells to undergo transendothelial migration (TEM) in vitro and form dynamic, F-actin–rich pseudopodia that penetrate capillary endothelial walls in vivo. Using multiple tumor cell lines, we show that the inability to form these membrane protrusions, which likely promote TEM and extravasation, can be explained by altered adhesion dynamics and impaired cell polarization that result when Rap1 activation or cycling is perturbed. Thus, targeting Rap1 could be a useful approach for reducing the metastatic dissemination of tumor cells that undergo active TEM. Cancer Res; 70(11); 4590–601. ©2010 AACR.
SuperiorIn vivoEfficacy of Afucosylated Trastuzumab in the Treatment of HER2-Amplified Breast Cancer
Authors: Junttila Teemu T. , Parsons Kathryn , Olsson Christine , Lu Yanmei , Xin Yan , Theriault Julie , Crocker Lisa , Pabonan Oliver , Baginski Tomasz , Meng Gloria , Totpal Klara , Kelley Robert F. , Sliwkowski Mark X.
Published: May 2010

The enhancement of immune effector functions has been proposed as a potential strategy for increasing the efficacy of therapeutic antibodies. Here, we show that removing fucose from trastuzumab (Herceptin) increased its binding to FcγRIIIa, enhanced antibody-dependent cell-mediated cytotoxicity, and more than doubled the median progression-free survival when compared with conventional trastuzumab in treating preclinical models of HER2-amplified breast cancer. Our results show that afucosylated trastuzumab has superior efficacy in treating in vivo models of HER2-amplified breast cancer and support the development of effector function–enhanced antibodies for solid tumor therapy. Cancer Res; 70(11); 4481–9. ©2010 AACR.
Immature Immunosuppressive CD14+HLA-DR−/low Cells in Melanoma Patients Are Stat3hi and Overexpress CD80, CD83, and DC-Sign
Authors: Poschke Isabel , Mougiakakos Dimitrios , Hansson Johan , Masucci Giuseppe V. , Kiessling Rolf
Published: May 2010

Myeloid-derived suppressor cells (MDSC) have emerged as key immune modulators in various tumor models and human malignancies, but their characteristics in humans remain to be unequivocally defined. In this study, we have examined circulating CD14+HLA-DR−/low MDSC in 34 advanced malignant melanoma (MM) patients. Their frequency is significantly increased and associated with disease activity. Contrary to the common notion that MDSC are a heterogeneous population of exclusively immature cells, we find the coexpression of markers associated with mature phenotype. We show for the first time the overexpression of CD80, CD83, and DC-Sign in human MDSC. Further, increased levels of signal transducer and activator of transcription 3 (Stat3), an important regulator in MDSC development and function, were noted in MM-MDSC. Stat3 was altered toward an active, phosphorylated state in the HLA-DR− population of CD14+ cells and was more reactive to activating stimuli in patients. Importantly, inhibition of Stat3 abolished their suppressive activity almost completely. The described MM-MDSC use arginase in conjunction with other yet undefined mechanisms to suppress CD4+ and CD8+ T cells. Several observations suggest a redox imbalance in MDSC and indicate an important role of Stat3-dependent oxidative stress in MDSC-mediated T-cell suppression. These results emphasize the diversity of MDSC in human cancer and provide potential targets for therapeutic interventions. Cancer Res; 70(11); 4335–45. ©2010 AACR.
Estrogen Receptor β1 Expression Is Regulated by miR-92 in Breast Cancer
Authors: Al-Nakhle Hakeemah , Burns Philip A. , Cummings Michele , Hanby Andrew M. , Hughes Thomas A. , Satheesha Sampoorna , Shaaban Abeer M. , Smith Laura , Speirs Valerie
Published: May 2010

Estrogen receptor β1 (ERβ1) downregulation occurs in many breast cancers, but the responsible molecular mechanisms remain unclear. Here, we report that levels of ERβ1 expression are negatively regulated by the microRNA miR-92. Expression analysis in a cohort of primary breast tumors confirmed a significant negative correlation between miR-92 and both ERβ1 mRNA and protein. Inhibition of miR-92 in MCF-7 cells increased ERβ1 expression in a dose-dependent manner, whereas miR-92 overexpression led to ERβ1 downregulation. Reporter constructs containing candidate miR-92 binding sites in the 3′-untranslated region (UTR) of ERβ1 suggested by bioinformatics analysis confirmed that miR-92 downregulated ERβ1 via direct targeting of its 3′-UTR. Our results define a potentially important mechanism for downregulation of ERβ1 expression in breast cancer. Cancer Res; 70(11); 4778–84. ©2010 AACR.
Persistent Cyclooxygenase-2 Inhibition Downregulates NF-κB, Resulting in Chronic Intestinal Inflammation in the Min/+ Mouse Model of Colon Tumorigenesis
Authors: Carothers Adelaide M. , Davids Jennifer S. , Damas Beatrice C. , Bertagnolli Monica M.
Published: May 2010

Cyclooxygenase-2 (COX-2) inhibition prevents adenoma formation in humans and mouse models of colon cancer. The selective COX-2 inhibitor celecoxib reduces COX-2 and prostaglandin E2 (PGE2) expression and adenomas in the intestine of Min/+ mice after treatment for several weeks, but prolonged treatment increases PGE2 production, resulting in drug-resistant tumor formation and transforming growth factor β (TGFβ)–dependent intestinal fibrosis. In this study, we examined pathways that regulate COX-2 expression and suppress chronic intestinal inflammation. We show that NF-κB signaling was inhibited in the ileum of Min/+ mice receiving long-term treatment with celecoxib. This effect was associated with inhibition of TGFβ-associated kinase-1 and IκB kinase α/β activities and reduced expression of the Toll-like receptor (TLR) 2 and TLR4 that enhance colonic barrier function. Additionally, we observed reduced activities of protein kinases c-Jun NH2-terminal kinase 1 and protein kinase A and transcription factor cyclic AMP–responsive element binding protein, regulators of COX-2 expression, which cross-talk with NF-κB. In ileum subjected to long-term celecoxib treatment, we noted relatively higher expression of COX-2, vascular endothelial growth factor, and interleukin-1β in Paneth cells, whereas NF-κB and COX-2 were more strongly expressed by an expanded population of stromal myofibroblasts. Our findings argue that celecoxib resistance is an acquired adaptation to changes in the crypt microenvironment that is associated with chronic intestinal inflammation and impaired acute wound-healing responsiveness. Cancer Res; 70(11); 4433–42. ©2010 AACR.
Type III IFN Interleukin-28 Mediates the Antitumor Efficacy of Oncolytic Virus VSV in Immune-Competent Mouse Models of Cancer
Authors: Wongthida Phonphimon , Diaz Rosa Maria , Galivo Feorillo , Kottke Timothy , Thompson Jill , Pulido Jose , Pavelko Kevin , Pease Larry , Melcher Alan , Vile Richard
Published: May 2010

Innate immune effector mechanisms triggered by oncolytic viruses may contribute to the clearance of both infected and uninfected tumor cells in immunocompetent murine hosts. Here, we developed an in vitro tumor cell/bone marrow coculture assay and used it to dissect innate immune sensor and effector responses to intratumoral vesicular stomatitis virus (VSV). We found that the type III IFN interleukin-28 (IL-28) was induced by viral activation of innate immune-sensing cells, acting as a key mediator of VSV-mediated virotherapy of B16ova melanomas. Using tumor variants which differentially express the IL-28 receptor, we showed that IL-28 induced by VSV within the tumor microenvironment sensitizes tumor cells to natural killer cell recognition and activation. These results revealed new insights into the immunovirological mechanisms associated with oncolytic virotherapy in immune-competent hosts. Moreover, they defined a new class of tumor-associated mutation, such as acquired loss of responsiveness to IL-28 signaling, which confers insensitivity to oncolytic virotherapy through a mechanism independent of viral replication in vitro. Lastly, the findings suggested new strategies to manipulate immune signals that may enhance viral replication, along with antitumor immune activation, and improve the efficacy of oncolytic virotherapies. Cancer Res; 70(11); 4539–49. ©2010 AACR.
The Fbw7 Tumor Suppressor Targets KLF5 for Ubiquitin-Mediated Degradation and Suppresses Breast Cell Proliferation
Authors: Zhao Dong , Zheng Han-Qiu , Zhou Zhongmei , Chen Ceshi
Published: May 2010

Fbw7 is a tumor suppressor frequently inactivated in cancers. The KLF5 transcription factor promotes breast cell proliferation and tumorigenesis through upregulating FGF-BP. The KLF5 protein degrades rapidly through the ubiquitin proteasome pathway. Here, we show that the Skp1-CUL1-Fbw7 E3 ubiquitin ligase complex (SCFFbw7) targets KLF5 for ubiquitin-mediated degradation in a GSK3β-mediated KLF5 phosphorylation–dependent manner. Mutation of the critical S303 residue in the KLF5 Cdc4 phospho-degrons motif (303SPPSS) abolishes the protein interaction, ubiquitination, and degradation by Fbw7. Inactivation of endogenous Fbw7 remarkably increases the endogenous KLF5 protein abundances. Endogenous Fbw7 suppresses the FGF-BP gene expression and breast cell proliferation through targeting KLF5 for degradation. These findings suggest that Fbw7 inhibits breast cell proliferation at least partially through targeting KLF5 for proteolysis. This new regulatory mechanism of KLF5 degradation may result in useful diagnostic and therapeutic targets for breast cancer and other cancers. Cancer Res; 70(11); 4728–38. ©2010 AACR.
Targeted Nanoparticles That Deliver a Sustained, Specific Release of Paclitaxel to Irradiated Tumors
Authors: Passarella Ralph J. , Spratt Daniel E. , van der Ende Alice E. , Phillips John G. , Wu Hongmei , Sathiyakumar Vasanth , Zhou Li , Hallahan Dennis E. , Harth Eva , Diaz Roberto
Published: May 2010

To capitalize on the response of tumor cells to XRT, we developed a controlled-release nanoparticle drug delivery system using a targeting peptide that recognizes a radiation-induced cell surface receptor. Phage display biopanning identified Gly-Ile-Arg-Leu-Arg-Gly (GIRLRG) as a peptide that selectively recognizes tumors responding to XRT. Membrane protein extracts of irradiated glioma cells identified glucose-regulated protein GRP78 as the receptor target for GIRLRG. Antibodies to GRP78 blocked the binding of GIRLRG in vitro and in vivo. Conjugation of GIRLRG to a sustained-release nanoparticle drug delivery system yielded increased paclitaxel concentration and apoptosis in irradiated breast carcinomas for up to 3 weeks. Compared with controls, a single administration of the GIRLRG-targeted nanoparticle drug delivery system to irradiated tumors delayed the in vivo tumor tripling time by 55 days (P = 0.0001) in MDA-MB-231 and 12 days in GL261 (P < 0.005). This targeting agent combines a novel recombinant peptide with a paclitaxel-encapsulating nanoparticle that specifically targets irradiated tumors, increasing apoptosis and tumor growth delay in a manner superior to known chemotherapy approaches. Cancer Res; 70(11); 4550–9. ©2010 AACR.
CXCL12 (SDF-1)/CXCR4 Pathway in Cancer
Authors: Teicher Beverly A. , Fricker Simon P.
Published: May 2010

Chemokines, small proinflammatory chemoattractant cytokines that bind to specific G-protein-coupled seven-span transmembrane receptors, are major regulators of cell trafficking and adhesion. The chemokine CXCL12 [stromal cell-derived factor-1 (SDF-1)] binds primarily to CXC receptor 4 (CXCR4; CD184). The binding of CXCL12 to CXCR4 induces intracellular signaling through several divergent pathways initiating signals related to chemotaxis, cell survival and/or proliferation, increase in intracellular calcium, and gene transcription. CXCR4 is expressed on multiple cell types including lymphocytes, hematopoietic stem cells, endothelial and epithelial cells, and cancer cells. The CXCL12/CXCR4 axis is involved in tumor progression, angiogenesis, metastasis, and survival. This pathway is a target for therapeutics that can block the CXCL12/CXCR4 interaction or inhibit downstream intracellular signaling. Clin Cancer Res; 16(11); 2927–31. ©2010 AACR.
Two-Stage Phase I Dose-Escalation Study of Intratumoral Reovirus Type 3 Dearing and Palliative Radiotherapy in Patients with Advanced Cancers
Authors: Harrington Kevin J. , Karapanagiotou Eleni M. , Roulstone Victoria , Twigger Katie R. , White Christine L. , Vidal Laura , Beirne Debbie , Prestwich Robin , Newbold Kate , Ahmed Merina , Thway Khin , Nutting Christopher M. , Coffey Matt , Harris Dean , Vile Richard G. , Pandha Hardev S. , DeBono Johann S. , Melcher Alan A.
Published: May 2010

Purpose: To determine the safety and feasibility of combining intratumoral reovirus and radiotherapy in patients with advanced cancer and to assess viral biodistribution, reoviral replication in tumors, and antiviral immune responses.

Experimental Design: Patients with measurable disease amenable to palliative radiotherapy were enrolled. In the first stage, patients received radiotherapy (20 Gy in five fractions) plus two intratumoral injections of RT3D at doses between 1 × 108 and 1 × 1010 TCID50. In the second stage, the radiotherapy dose was increased (36 Gy in 12 fractions) and patients received two, four, or six doses of RT3D at 1 × 1010 TCID50. End points were safety, viral replication, immunogenicity, and antitumoral activity.

Results: Twenty-three patients with various solid tumors were treated. Dose-limiting toxicity was not seen. The most common toxicities were grade 2 (or lower) pyrexia, influenza-like symptoms, vomiting, asymptomatic lymphopenia, and neutropenia. There was no exacerbation of the acute radiation reaction. Reverse transcription-PCR (RT-PCR) studies of blood, urine, stool, and sputum were negative for viral shedding. In the low-dose (20 Gy in five fractions) radiation group, two of seven evaluable patients had a partial response and five had stable disease. In the high-dose (36 Gy in 12 fractions) radiation group, five of seven evaluable patients had partial response and two stable disease.

Conclusions: The combination of intratumoral RT3D and radiotherapy was well tolerated. The favorable toxicity profile and lack of vector shedding means that this combination should be evaluated in newly diagnosed patients receiving radiotherapy with curative intent. Clin Cancer Res; 16(11); 3067–77. ©2010 AACR.
Structural Basis on the Dityrosyl-Diiron Radical Cluster and the Functional Differences of Human Ribonucleotide Reductase Small Subunits hp53R2 and hRRM2
Authors: Zhou Bingsen , Su Leila , Yuan Yate-Ching , Un Frank , Wang Norby , Patel Madhukar , Xi Bixin , Hu Shuya , Yen Yun
Published: May 2010

Ribonucleotide reductase (RNR) is an enzyme for the de novo conversion of ribonucleotides to deoxyribonucleotides. The two human RNR small subunits hRRM2 and hp53R2 share 83% sequence homology but show distinct expression patterns and function. Structural analyses of the oxidized form of hRRM2 and hp53R2 indicate that both proteins contain a conserved Gln127-hp53R2/Gln165-hRRM2 close to the dinuclear iron center and the essential tyrosine residue Tyr124-hp53R2/Tyr162-hRRM2 forms hydrogen bonds with the tyrosine and iron ligands, implying a critical role for the glutamine residue in assembling the dityrosyl-diiron radical cofactor. The present work also showed that Tyr221 in hRRM2, which is replaced by Phe183 in hp53R2, forms a hydrogen bond with Tyr162 to extend the hydrogen bond network from Gln165-hRRM2. Mutagenesis and spectroscopic experiments suggested that the tyrosine-to-phenylalanine switch at Phe183-hp53R2/Tyr221-hRRM2 could lead to differences in radical generation or enzymatic activity for hp53R2 and hRRM2. This study correlates the distinct catalytic mechanisms of the small subunits hp53R2 and hRRM2 with a hydrogen-bonding network and provides novel directions for designing and developing subunit-specific therapeutic agents for human RNR enzymes. Mol Cancer Ther; 9(6); 1669–79. ©2010 AACR.
p53-Independent Apoptosis by Benzyl Isothiocyanate in Human Breast Cancer Cells Is Mediated by Suppression of XIAP Expression
Authors: Kim Su-Hyeong , Singh Shivendra V.
Published: May 2010

We have shown previously that cruciferous vegetable constituent benzyl isothiocyanate (BITC) suppresses viability of cultured MCF-7 and MDA-MB-231 human breast cancer cells and retards mammary cancer development in MMTV-neu mice by causing apoptosis, but the mechanism of cell death is not fully understood. We now show that whereas p53 is dispensable for BITC-induced cell death, proapoptotic response to this promising chemopreventive agent is mediated by suppression of X-linked inhibitor of apoptosis (XIAP) protein expression. The BITC treatment increased levels of total and Ser15-phosphorylated p53 protein in MCF-7 cells, but the proapoptotic response to this agent was maintained even after knockdown of the p53 protein level. Exposure of MCF-7 and MDA-MB-231 cells to BITC resulted in a marked decrease in protein level of XIAP as early as 8 hours after treatment. Ectopic expression of XIAP conferred statistically significant protection against BITC-mediated cytoplasmic histone-associated apoptotic DNA fragmentation in both cell lines. Moreover, inhibition of MDA-MB-231 cell growth in vivo in female athymic mice by BITC administration correlated with a modest but statistically significant decrease in XIAP protein level in the tumor xenograft. The BITC treatment also resulted in induction as well as nuclear translocation of survivin only in the MCF-7 cells. The BITC-induced apoptosis was modestly but statistically significantly augmented by RNA interference of survivin in MCF-7 cells. In conclusion, the present study provides novel insight into the molecular circuitry of BITC-induced apoptosis to indicate suppression of XIAP expression as a critical mediator of this process. Cancer Prev Res; 3(6); 718–26. ©2010 AACR.
TAS2R38 Genotypes and Phenylthiocarbamide Bitter Taste Perception in a Population of Young Adults
Authors: Khataan Nora H. , Stewart Lindsay , Brenner Darren M. , Cornelis Marilyn C. , El-Sohemy Ahmed
Published: May 2010

Background/Aims:TAS2R38 belongs to the TAS2R bitter taste receptor gene family and polymorphisms are associated with differences in bitter taste perception of phenylthiocarbamide (PTC) and 6-n-propylthiouracil (PROP). The objectives were to test whether the genotype-phenotype relationship varies between ethnocultural groups and whether a filter paper method can predict TAS2R38 genotype. Methods: Subjects (n = 911) aged 20–29 rated the intensity of a PTC-containing filter paper on a scale of 1–9, from ‘not at all bitter’ to ‘extremely bitter’. The TAS2R38 A49P polymorphism was detected by real-time PCR. Results: The overall frequency of the 49A variant was 0.47 and the AA, AP, and PP genotype distribution was 0.24, 0.45 and 0.31, respectively. Those with AA, AP, or PP genotypes had mean bitterness rating scores of 2.3, 4.9, and 5.9, respectively. The 49A variant frequency was significantly lower among Asians (28%) as compared to Caucasians (55%), South Asians (60%) or others (56%) (p < 0.0001). The genotype-phenotype relationship was similar across ethnocultural groups, however, a stronger genotype-phenotype association was observed among women as compared to men. Conclusion: Our findings show that a simple PTC filter paper tasting method strongly predicts TAS2R38 genotype, and that the genotype-phenotype association is similar among ethnocultural groups.
Frequency and Determinants of Poststroke Cognitive Impairment at Three and Twelve Months in Chile
Authors: Delgado Carolina , Donoso Archibaldo , Orellana Patricia , Vásquez Carolina , Díaz Violeta , Behrens María Isabel
Published: May 2010

Background: A higher risk of poststroke cognitive impairment (CI) has been reported in Hispanics in a US cohort but has not been systematically studied in Latin America. Objectives: Our purpose was to investigate the frequencies and determinants of poststroke CI in the hispano-mestizo population of Santiago, Chile. Methods: A prospective study of hospitalized patients aged >60 years admitted with an ischemic or hemorrhagic stroke was conducted. The cognitive status was determined at 3 and 12 months after the stroke by informant questionnaires, neuropsychological testing and clinical diagnosis. Cardiovascular risk factors, brain imaging and stroke features were analyzed using regression models to establish determinants for poststroke CI. Results: A total of 164 patients (mean age = 72 ± 7.5 years) were recruited. Out of 122 patients (74%) evaluated at 3 months, 81 (66%) had CI. Out of 101 patients (62%) evaluated at 12 months, 39 (39%) had CI no dementia, and 22 (22%) were demented. The new-onset dementia frequency at 1 year was 16%. Independent determinants for dementia were higher functional impairment at hospital egress (OR = 4.0), left-hemisphere large-vessel infarction (OR = 6.9) and a larger amount of white matter changes (OR = 1.3). Conclusions: In this first study on poststroke CI in Latin America, the frequencies and determinants of poststroke CI were similar to those in other cohorts of different ethnic origin.
Interaction of the Anti-Inflammatory Annexin A1 Protein and Tacrolimus Immunosuppressant in the Renal Function of Rats
Authors: Araujo Leandro P. , Truzzi Renata R. , Mendes Gloria E. , Luz Marcus A.M. , Burdmann Emmanuel A. , Oliani Sonia M.
Published: May 2010

Background: Tacrolimus (FK) is currently widely used in transplant immunosuppression and the treatment of autoimmune diseases. However, FK induces nephrotoxicity which is characterized by functional and structural renal injury. The ubiquitous protein annexin A1 (ANXA1) has potent anti-inflammatory effects and protects against ischemia/reperfusion injury. We investigated the effects of exogenous ANXA1 treatment in an experimental model of acute FK nephrotoxicity. Methods: Munich-Wistar rats received a low-salt diet for 1 week and were randomized to treatment with ANXA1 (Ac2-26 peptide 0.5 mg/kg/day s.c.), FK (6 mg/kg/day p.o.), association (FK+ANXA1) and vehicles (1 ml/kg/day) for 7 days. Results: FK induced a significant decrease in glomerular filtration rate and renal blood flow, and a significant increase in renal vascular resistance. In addition, FK caused extensive acute tubule-interstitial damage and an increase in anti-inflammatory ANXA1 expression in renal tissue. Exogenous ANXA1 treatment reduced FK-induced tubular dilatation and macrophage infiltration. For the first time, we observed that FK augmented ANXA1 expression in renal tissue. Conclusion: Exogenous ANXA1 treatment partially protected against FK-induced tubular injury and macrophage infiltration, and may be targeted in renal intervention strategies.
Role of the Intrarenal Renin-Angiotensin-Aldosterone System in Chronic Kidney Disease
Authors: Siragy Helmy M. , Carey Robert M.
Published: May 2010

The existence of local or tissue-based renin-angiotensin-aldosterone systems (RAAS) is well documented and has been implicated as a key player in the pathogenesis of cardiovascular and renal diseases. The kidney contains all elements of the RAAS, and intrarenal formation of angiotensin II not only controls glomerular hemodynamics and tubule sodium transport, but also activates a number of inflammatory and fibrotic pathways. Experimental and clinical studies have shown that the intrarenal RAAS is activated early in diabetic nephropathy, the leading cause of chronic kidney disease (CKD). Although angiotensin-converting enzyme inhibitors and angiotensin receptor blockers decrease the rate of decline in kidney function in patients with diabetic and non-diabetic nephropathy, many patients still progress to end-stage renal disease or die from cardiovascular events. There is still a clear need for additional strategies to block the RAAS more effectively to reduce progression of CKD. The focus of this paper is to review the importance of the intrarenal RAAS in CKD and recent findings in renin-angiotensin biology pertinent to the kidney. We also discuss additional strategies to inhibit the RAAS more effectively and the potential impact of direct renin inhibition on the prevention and management of CKD.
Surveillance and Modification of Immunosuppression Minimizes BK Virus Nephropathy
Authors: Bennett William M. , Meyer Leslie , Ridenour Jennifer , Batiuk Thomas D.
Published: May 2010

Background/Aims: Infection of a transplanted kidney with the polyomavirus, BK, is associated with poor allograft survival. Methods: In an attempt to prevent this transplant complication, we studied 144 consecutive transplant recipients for the presence of BK infection with plasma and urine PCR testing at 1, 2, 3, 6 and 12 months. Viruria alone was followed by serial studies. If plasma PCR became positive at >2.6 log copies, mycophenolate was reduced until there was no detectable plasma viral load. Results: Urine PCR was positive in 34 (24%), while plasma PCR turned positive in 22 cases (15%). No patients developed viremia with <6.8 log copies in the urine. Viremia resolved within 3 months or less in 20 of 22 patients after reduction of immunosuppression. Surveillance biopsies at 2 and 6 months revealed no BK nephropathy. Eight patients had acute rejection during reduced immunosuppression; however, all of these reversed with pulse steroids. Patient and graft survival at 1 year was 99 and 98%, respectively. Use of the cell-mediated immunity assay (ImmuKnow®, Columbia, Md., USA) was not useful in identifying infected patients. Conclusion: Active surveillance for BK virus by urine/plasma PCR with prompt reduction in immunosuppression can prevent BK nephropathy.
Excess Mortality Rates for Estimating the Non-Fatal Burden of Stroke in Western Australia: A Data Linkage Study
Authors: Katzenellenbogen Judith M. , Vos Theo , Somerford Peter , Begg Stephen , Semmens James B. , Codde James P.
Published: May 2010

Background: Non-fatal stroke burden measured in Years Lived with Disability (YLD) requires valid estimates of stroke case fatality to allow modelling of disease duration. In the model, case fatality can be calculated from the absolute risk of mortality in cases in excess of that in the non-diseased. Aims: Our purpose was to estimate excess mortality rates in 28-day survivors of stroke in Western Australia and to evaluate differentials in survival by stroke type, age and time since the first stroke event. Method: Excess mortality among prevalent (first-ever plus existing) survivors was estimated from linked hospital and mortality data. Changes in excess mortality over time were calculated over a 6-year period. Results: Excess mortality increased with age for both males (21 per 1,000 in the 15- to 54-year to 109 per 1,000 in the ≧85-year age group) and females (16 and 122 per 1,000 for the 15- to 54-year and ≧85-year groups, respectively). Survival by stroke sub-types differed at ages <55 years but not >55 years. During the first year excess mortality was markedly higher, after which it was relatively constant for each age group. The assumption of constant rather than changing excess mortality in 28-day survivors of stroke had minimal effect on estimates of duration. Conclusion: Measures of excess mortality in prevalent survivors have not previously been available for estimating YLD for stroke. An analysis of all stroke types combined is not likely to substantially bias estimates of non-fatal stroke burden nor is an assumption of constant excess mortality for survivors.
Intracranial Internal Carotid Artery Calcification: A Representative for Cerebral Artery Calcification and Association with White Matter Hyperintensities
Authors: Chung Pil-Wook , Park Kwang-Yeol , Moon Heui-Soo , Kim Yong-Bum , Youn Young Chul , Byun Jun Soo , Kwon Oh-Sang
Published: May 2010

Background: Our aim was to investigate the distribution pattern of cerebral artery calcification and its association with white matter hyperintensities (WMH). Methods: We identified 159 consecutive patients with acute ischemic stroke. Calcifications of cerebral arteries and WMH were graded. Results: Cerebral artery calcification was found in 137 patients (86.2%). The intracranial internal carotid artery (I-ICA) was the most frequently affected artery with calcification (76.7%) and moderate-to-severe calcification (38.1%). Spearman’s rank test revealed that the grade of I-ICA calcification was correlated with those of periventricular WMH (r = 0.417, p < 0.001) and deep WMH (r = 0.388, p < 0.001). The adjusted ORs of I-ICA were 2.62 (p <0.05) for periventricular WMH and 3.25 (p <0.05) for deep WMH. Conclusions: Cerebral artery calcification is common in patients with ischemic stroke. I-ICA is the most frequently and most severely affected cerebral artery and its calcification is associated with WMH.
Efficacy of Fluoride Compounds and Stannous Chloride as Erosion Inhibitors in Dentine
Authors: Ganss C. , Lussi A. , Sommer N. , Klimek J. , Schlueter N.
Published: May 2010

The aim of this study was to evaluate the anti-erosive effects of different fluoride compounds and one tin compound in the context of the complex pathohistology of dentine erosion, with particular emphasis on the role of the organic portion. Samples were subjected to two experiments including erosive acid attacks (0.05 molar citric acid, pH 2.3; 6 × 2 min/day) and applications (6 × 2 min/day) of the following test solutions: SnCl2 (815 ppm Sn), NaF (250 ppm F), SnF2 (250 ppm F, 809 ppm Sn), amine fluoride (AmF, 250 ppm F), AmF/NaF (250 ppm F), and AmF/SnF2 (250 ppm F, 409 ppm Sn). The demineralised organic fraction was enzymatically removed either at the end of the experiment (experiment 1) or continuously throughout the experiment (experiment 2). Tissue loss was determined profilometrically after 10 experimental days. In experiment 1, the highest erosive tissue loss was found in the control group (erosion only); the AmF- and NaF-containing solutions reduced tissue loss by about 60%, reductions for SnCl2, AmF/SnF2, and SnF2 were 52, 74 and 89%, respectively. In experiment 2, loss values generally were significantly higher, and the differences between the test solutions were much more distinct. Reduction of tissue loss was between 12 and 34% for the AmF- and NaF-containing preparations, and 11, 67 and 78% for SnCl2, AmF/SnF2, and SnF2, respectively. Stannous fluoride-containing solutions revealed promising anti-erosive effects in dentine. The strikingly different outcomes in the two experiments suggest reconsidering current methodologies for investigating anti-erosive strategies in dentine.
In situ Remineralisation of Enamel and Dentin after the Use of an Amine Fluoride Mouthrinse in Addition to Twice Daily Brushings with Amine Fluoride Toothpaste
Authors: Laheij A.M.G.A. , van Strijp A.J.P. , van Loveren C.
Published: May 2010

It is often claimed that 3 fluoride moments a day significantly reduce the caries risk compared to 2 daily fluoride moments. However, previous research is not conclusive. Therefore, the aim of this study was to compare the effect on lesion progression of 2 versus 3 fluoride moments a day. A double-blind, randomized, cross-over in situ experiment was designed. The experiment comprised 2 in situ periods of 3 weeks with a washout period of 3 weeks in between. Sixteen participants wore an enamel and a dentine specimen with a preformed lesion placed buccally in their partial prosthesis. The participants brushed twice a day with a 1,400 ppm F (amine fluoride) toothpaste and rinsed once a day with either 250 ppm F (amine F/NaF) or a placebo rinse. At the end of the experiment the specimens were retrieved for fluoride analysis and the assessment of integrated mineral loss with transversal microradiography. The fluoride analysis showed a statistically significant increase in structurally bound fluoride in dentine, but not in enamel, when comparing the fluoride mouthrinse group with the placebo rinse group. The amounts of loosely bound, KOH-soluble fluoride were not different between both groups neither for enamel nor for dentine. In dentine IML gain was significantly (p < 0.05) higher for the fluoride mouthrinse group than for the placebo mouthrinse group. In enamel no statistically significant differences in IML gain were found. For dentine a third fluoride moment may be beneficial in enhancing remineralisation, even under the remineralising conditions as in this study.
Association of Oral Candidal Carriage with Dental Caries in Children
Authors: Raja M. , Hannan A. , Ali K.
Published: May 2010

Introduction: Oral candidiasis is one of the most common opportunistic oral fungal infections. Oral candidal carriage in schoolchildren is a subject of increasing interest worldwide and has recently been associated with increased caries incidence in children. Aims: This study was carried out to identify association between oral candidal carriage in children and dental caries. Subjects and Methods: One hundred subjects with an age range between 6 and 12 years were included in this study. The subjects were distributed equally into two groups, i.e., study (caries-positive) and control (caries-free) groups. Oral hygiene index and DMFT/dmft scores were recorded for each subject. Sampling for Candida was carried out using intraoral swabs and concentrated oral rinse. Sabouraud dextrose agar containing 0.1 mg/ml of chloramphenicol was used as the primary culture medium. Candida was identified by employing API-20C AUX and germ tube formation tests. Results: The subjects in the caries-positive group showed a high frequency of oral candidal carriage compared to the caries-free subjects and the results were statistically very significant (p < 0.01). The odds ratio was 67.37, implying a high caries risk with positive oral candidal carriage (95% CI 14–323). Discussion: The findings of this study underscore the possible association of Candida with dental caries. Conclusions: The occurrence of dental caries in children with mixed dentition is positively correlated with the frequency of oral candidal carriage.
Octenidine Dihydrochloride, a Modern Antiseptic for Skin, Mucous Membranes and Wounds
Authors: Hübner N.-O. , Siebert J. , Kramer A.
Published: May 2010

Octenidine dihydrochloride (octenidine) was introduced for skin, mucous membrane and wound antisepsis more than 20 years ago. Until now, a wealth of knowledge has been gained, including in vitro and animal studies on efficacy, tolerance, safety and clinical experience both from case reports and prospective controlled trials. Nowadays, octenidine is an established antiseptic in a large field of applications and represents an alternative to older substances such as chlorhexidine, polyvidone-iodine or triclosan. The review is based on the current literature and unpublished original data as well.