bluuebunny/crossref_metadata_2025 huggingface dataset:
Distribution, geometry and palaeogeography of the Frasnian (Late Devonian) reef complexes of Banks Island, NWT, western arctic, Canada
Authors: Copper P. , Edinger E.
Published: April 2009

Following the collapse of the >2000 km long Givetian (Middle Devonian) Inuitian/Ellesmere carbonate platform factory in arctic Canada, within the 0° to 10° equatorial palaeolatitudes north, the only Frasnian reefs in high arctic Canada retreated westwards, confined to northeastern Banks Island. These reefs, numbering well over 130, and dominated by corals and stromatoporoid sponges, were spread over
c
. 5000 km
2
, within the 220 m thick Mercy Bay Formation. Reefs were developed at four different stratigraphic levels (termed the A, B, C and D levels) during early and middle Frasnian time, periodically smothered by intervening siliciclastics during sea-level lowstands, and were finally buried by thick siliciclastic sands, silts and muds derived from the east during the late Frasnian. The Frasnian–Famennian boundary is masked within continental, deltaic facies bearing plant remains. The Banks reef and carbonate succession is preserved as horizontally stratified,
in situ
limestones, with the succeeding reefs backsteppping towards the east, in response to cyclic Frasnian sea-level rises. The Banks reefs were developed on the distal lobes of a megadelta, periodically covered by extensive lowland forests that stretched more than 2000 km west from Greenland and Ellesmere Island. Reefs are three-dimensionally preserved, unaffected by diagenesis, dolomitization, major tectonics, vegetative cover or glaciation, and thus display some of the finest, pristine Late Devonian reef complexes known. The reefs represent a range of geometries, from small circular to oval patch reefs a few tens of metres in diameter and <5–10 m thick, to larger prominent tabular (‘reef platform’ style) and domal reef structures >2 km across, or 300–400 m diameter, and 40–60 m thick. Coral and stromatoporoid-rich biostromes also formed a significant part of the reef seascape. Morphology of the reefs leads to an estimation of penecontemporaneous relief of 10–20 m above the prevailing sea floor, with reef-core facies generally dominated by stromatoporoid sponges, and flank facies by a variable mix of colonial tabulate and rugose corals, as well as stromatoporoids. Bryozoans and calcimicrobialites were rare, in contrast to other Late Devonian reefs such as those of the Canning Basin (Australia), or the Guangxi platform (South China); mudmound reef facies, such as seen in Belgium were limited.
Devonian (Emsian to Frasnian) crinoids of the Dra Valley, western Anti-Atlas Mountains, Morocco
Authors: Webster G. D. , Becker R. T.
Published: April 2009

Emsian to Frasnian crinoids are described from pelagic facies of the eastern part of the Dra Valley (Tata area), western Anti-Atlas Mountains, southern Morocco. The crinoids show only minor relationship with previously described crinoids from the Tafilalt and Ma'der areas of the eastern Anti-Atlas. The differences are judged to reflect the different environments of these areas. The Dra Valley hexacrinids show greater affinity with European faunas, whereas the amabilicrinids show greater affinity with North American taxa. New taxa described are
Hexacrinites chenae
sp. nov.,
Dracrinus crenulatus
gen. and sp. nov.,
Coquinacrinus revimentus
gen. and sp. nov, and
Embolocrinus quadruus
gen. and sp. nov.
A re-evaluation of Famennian echinoderm diversity: implications for patterns of extinction and rebound in the Late Devonian
Authors: Waters J. A. , Webster G. D.
Published: April 2009

Critical to understanding long-term trends in diversity is a dataset that is both worldwide in scope and based on a sound taxonomic foundation. In this paper we re-evaluate the Famennian (Late Devonian) echinoderm dataset, which has changed radically in the past decade, and reinterpret patterns of Late Devonian echinoderm extinction and rebound based on these new data. Historically, Famennian (Late Devonian) and earliest Carboniferous echinoderms have been poorly known on a global basis leading to interpretations of prolonged rebound from the Devonian extinction events. Recent discoveries of abundant and diverse Famennian echinoderm faunas from northwestern China, Colorado, Australia and Iran, together with re-examination of previously known echinoderm faunas from Germany and England, have altered drastically our understanding of the patterns of extinction and rebound of Famennian and earliest Carboniferous echinoderm communities. Overall, Famennian echinoderm diversity at the generic level is nearly five times greater than reported in the 2002 Sepkoski compilation, and familial level diversity is more than seven times greater than previously thought. Despite the increases in diversity, Famennian echinoderm faunas show a reduced diversity of camerate crinoids that typify both Middle Devonian and Lower Mississippian faunas and portend the rise of cladid crinoid diversity later in the Carboniferous. Individual Famennian faunas are numerically dominated by blastoids, which also portends trends seen at various times later in the Palaeozoic. In general, we are able to recognize the following trends. Rebound from the Late Devonian extinction events in echinoderms was more rapid than previously thought, but seems to be concentrated in Asia. Palaeogeographically Famennian echinoderms can be grouped into two broad regions: one includes China, Australia and Iran, all of which bordered the Palaeotethys; the other includes regions from Laurussia (Europe and North America) and northern Africa (Morocco).
Dynamics of glutathione and ophthalmate traced with2H-enriched body water in rats and humans
Authors: Kombu Rajan S. , Zhang Guo-Fang , Abbas Rime , Mieyal John J. , Anderson Vernon E. , Kelleher Joanne K. , Sanabria Juan R. , Brunengraber Henri
Published: April 2009

We developed a LC-MS-MS assay of the2H labeling of free glutathione (GSH) and bound glutathione [GSSR; which includes all DTT-reducible forms, primarily glutathione disulfide (GSSG) and mixed disulfides with proteins] and ophthalmate (an index of GSH depletion) labeled from2H-enriched body water. In rats whose body water was 2.5%2H enriched for up to 31 days, GSH labeling follows a complex pattern because of different rates of labeling of its constitutive amino acids. In rats infused with [13C2,15N-glycine]glutathione, the rate of appearance of plasma GSH was 2.1 μmol·min−1·kg−1, and the half-life of plasma GSH/GSSR was 6–8 min. In healthy humans whose body fluids were 0.5%2H enriched, the2H labeling of GSH/GSSR and ophthalmate can be precisely measured after 4 h, with GSH being more rapidly labeled than GSSR. Since plasma GSH/GSSR derives mostly from liver, this technique opens the way to 2) probe noninvasively the labeling pattern and redox status of the liver GSH system in humans and 2) assess the usefulness of ophthalmate as an index of GSH depletion.
Neutral lipid storage disease: genetic disorders caused by mutations in adipose triglyceride lipase/PNPLA2orCGI-58/ABHD5
Authors: Schweiger Martina , Lass Achim , Zimmermann Robert , Eichmann Thomas O. , Zechner Rudolf
Published: April 2009

Neutral lipid storage disease (NLSD) is a group of autosomal recessive disorders characterized by the excessive accumulation of neutral lipids in multiple tissues. Recently, two genes, adipose triglyceride lipase ( ATGL/ PNPLA2) and comparative gene identification-58 ( CGI-58/ABHD5), have been shown to cause NLSD. ATGL specifically hydrolyzes the first fatty acid from triacylglycerols (TG) and CGI-58/ABHD5 stimulates ATGL activity by a currently unknown mechanism. Mutations in both the ATGL and the CGI-58 genes are associated with systemic TG accumulation, yet the resulting clinical manifestations are not identical. Patients with defective ATGL function suffer from more severe myopathy (NLSDM) than patients with defective CGI-58 function. On the other hand, CGI-58 mutations are always associated with ichthyosis (NLSDI), which was not observed in patients with defective ATGL function. These observations indicate an ATGL-independent function of CGI-58. This review summarizes recent findings with the goal of relating structural variants of ATGL and CGI-58 to functional consequences in lipid metabolism.
Colocalization of ghrelinO-acyltransferase and ghrelin in gastric mucosal cells
Authors: Sakata Ichiro , Yang Jing , Lee Charlotte E. , Osborne-Lawrence Sherri , Rovinsky Sherry A. , Elmquist Joel K. , Zigman Jeffrey M.
Published: April 2009

Ghrelin is a peptide hormone with many known functions, including orexigenic, blood glucose-regulatory, and antidepressant actions, among others. Mature ghrelin is unique in that it is the only known naturally occurring peptide to be posttranslationally modified by O-acylation with octanoate. This acylation is required for many of ghrelin's actions, including its effects on promoting increases in food intake and body weight. GOAT (ghrelin O-acyltransferase), one of 16 members of the MBOAT family of membrane-bound O-acyltransferases, has recently been identified as the enzyme responsible for catalyzing the addition of the octanoyl group to ghrelin. Although the initial reports of GOAT have localized its encoding mRNA to tissues known to contain ghrelin, it is as yet unclear whether the octanoylation occurs within ghrelin-producing cells or in neighboring cells. Here, we have performed dual-label histochemical analysis on mouse stomach sections and quantitative PCR on mRNAs from highly enriched pools of mouse gastric ghrelin cells to demonstrate a high degree of GOAT mRNA expression within ghrelin-producing cells of the gastric oxyntic mucosa. We also demonstrate that GOAT is the only member of the MBOAT family whose expression is highly enriched within gastric ghrelin cells and whose whole body distribution mirrors that of ghrelin.
Involvement of NADPH oxidase in oxidized LDL-induced upregulation of heat shock factor-1 and plasminogen activator inhibitor-1 in vascular endothelial cells
Authors: Zhao Ruozhi , Ma Xiuli , Xie Xueping , Shen Garry X.
Published: April 2009

Plasminogen activator inhibitor-1 (PAI-1) is implicated in thrombogenesis, inflammation, and extracellular matrix remodeling. Previous studies indicated that oxidized low-density lipoprotein (LDL) stimulated the generation of PAI-1 in vascular endothelial cells (EC). The present study demonstrated that LDL oxidized by copper, iron, or 3-morpholinosydnonimine increased the expression of NADPH oxidase (NOX) 2, PAI-1, and heat shock factor-1 (HSF1) in human umbilical vein EC or coronary artery EC compared with LDL or vehicle. Diphenyleneiodonium, a NOX inhibitor, prevented the increases of the expression of HSF1 and PAI-1 in EC induced by oxidized LDLs. Small-interference RNA (siRNA) for p22phox, an essential subunit of NOX, prevented oxidized LDL-induced expression of NOX2, HSF1, and PAI-1 in EC. HSF1 siRNA inhibited oxidized LDL-induced expression of PAI-1 and HSF1, but not NOX2, in EC. The binding of HSF1 to PAI-1 promoter and the activity of PAI-1 promoter in EC were enhanced by oxidized LDL. Butylated hydroxytulene, a potent antioxidant, inhibited oxidized LDL-induced release of hydrogen peroxide (H2O2) and the expression of NOX2, HSF1, and PAI-1 in EC. Treatment with H2O2 increased the abundance of NOX2, HSF1, and PAI-1 in EC. The results of the present study indicate that oxidized LDL-induced expression of NOX may lead to the elevated release of reactive oxygen species, the activation of HSF1, and the enhancement of the transcription of PAI-1 gene in cultured vascular EC.
Prenatal programming of rat thick ascending limb chloride transport by low-protein diet and dexamethasone
Authors: Dagan Amit , Habib Sabeen , Gattineni Jyothsna , Dwarakanath Vangipuram , Baum Michel
Published: April 2009

Prenatal administration of dexamethasone and a low-protein diet has been shown to result in hypertension in the offspring when they are adults. The cause for the hypertension is unknown. The purpose of this study was to examine whether there was prenatal programming of thick ascending limb transport. Rats were administered either dexamethasone for 4 days (0.2 mg/kg body wt) by intraperitoneal injection daily between the 15th and 18th day of gestation, or they were fed a low-protein diet (6% protein) or an isocaloric normal protein diet (20% protein) from day 12 gestation until birth. The offspring were studied as adults. Prenatal dexamethasone and dietary protein deprivation resulted in an increase in blood pressure. Offspring of mothers fed a low-protein diet had an increase in medullary but not cortical bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2) protein abundance ( P < 0.01). There was not a statistically significant increase in medullary NKCC2 by prenatal dexamethasone ( P = 0.07). Both prenatal administration of dexamethasone and a low-protein diet resulted in an increase in medullary thick ascending limb chloride transport compared with control (298 ± 33 pmoles·mm−1·min−1, 280 ± 26 pmoles·mm−1·min−1, and 191 ± 21 pmoles·mm−1·min−1, respectively P < 0.05). There was a higher lumen-positive transepithelial potential difference in the prenatal dexamethasone and low-protein group compared with control as well. Administration of furosemide for 24 h resulted in a decrease in blood pressure in the low-protein group but not the control group. This study demonstrates that insults administered to the fetus can program altered sodium transport. Increased tubular sodium transport is a likely cause for the hypertension by prenatal programming.
Dynamical Foundations of the Neural Circuit for Bayesian Decision Making
Authors: Morita Kenji
Published: April 2009

On the basis of accumulating behavioral and neural evidences, it has recently been proposed that the brain neural circuits of humans and animals are equipped with several specific properties, which ensure that perceptual decision making implemented by the circuits can be nearly optimal in terms of Bayesian inference. Here, I introduce the basic ideas of such a proposal and discuss its implications from the standpoint of biophysical modeling developed in the framework of dynamical systems.
Equine thrombospondin II and secreted protein acidic and cysteine-rich in a model of normal and pathological wound repair
Authors: Miragliotta Vincenzo , Raphaël Kevin , Ipiña Zoë , Lussier Jacques G. , Theoret Christine L.
Published: April 2009

Wound healing in horses is complicated, particularly when wounds are on the limb. The objectives of this study were to clone equine thrombospondin II (THBS2) and secreted protein acidic and cysteine-rich (SPARC) cDNAs and to compare the spatiotemporal expression of mRNAs and proteins during repair of body and limb wounds. These molecules were targeted in view of their potential biological contribution to angiogenesis, which is exacerbated during the repair of limb wounds in horses. Cloning was achieved by screening size-selected cDNA libraries previously derived from 7-day-old wounds. Expression was studied in unwounded skin and in samples from 1, 2, 3, 4, and 6 wk old wounds of the body and limb. Temporal gene expression was determined by semiquantitative RT-PCR, while protein expression was mapped immunohistochemically. The temporal pattern of expression for both genes was similar; wounding caused immediate upregulation of mRNA, which did not return to baseline by the end of the study, and overexpression was noted in body relative to limb wounds. Immunostaining for THBS2 and SPARC was induced by wounding, though no differences in stain location or intensity were detected between body and limb wounds. This study is the first to characterize equine cDNA for THBS2 and SPARC and to document mRNA expression over the different phases of repair. THBS2 and SPARC might modulate angiogenesis during wound healing in the horse, which could protect against the disproportionate fibroplasia commonly afflicting limb wounds and leading to the development of exuberant granulation tissue.
Regulation of Secondary Antigen-Specific CD8+ T-Cell Responses by Natural Killer T Cells
Authors: Hong Changwan , Lee Hyunji , Park Yoon-Kyung , Shin Junghoon , Jung Sundo , Kim Hoyeon , Hong Seokmann , Park Se-Ho
Published: April 2009

The physiologic function of natural killer T (NKT) cells in adaptive immunity remains largely unknown because most studies have used NKT cell agonists. In the present study, the role of NKT cells during the secondary effector phase was investigated separately from the primary immunization phase via adoptive transfer of differentiated effector T cells into naive recipients. We found that secondary antitumor CD8+ T-cell responses were optimal when NKT cells were present. Tumor-specific CD8+ effector T cells responded less strongly to tumor cell challenge in NKT cell–deficient recipients than in recipients with intact NKT cells. NKT cell–mediated enhancement of the secondary antitumor CD8+ T-cell response was concurrent with increased number and activity of tumor-specific CD8+ T cells. These findings provide the first demonstration of a direct role for NKT cells in the regulation of antigen-specific secondary T-cell responses without the use of exogenous NKT cell agonists such as α-galactosylceramide (α-GalCer). Furthermore, forced activation of NKT cells with α-GalCer during the secondary immune response in suboptimally immunized animals enhanced otherwise poor tumor rejection responses. Taken together, our findings strongly emphasize the importance of NKT cells in secondary CD8+ T-cell immune responses. [Cancer Res 2009;69(10):4301–8]
Disruption of the E2 Gene Is a Common and Early Event in the Natural History of Cervical Human Papillomavirus Infection: A Longitudinal Cohort Study
Authors: Collins Stuart I. , Constandinou-Williams Christothea , Wen Kaisheng , Young Lawrence S. , Roberts Sally , Murray Paul G. , Woodman Ciaran B.J.
Published: April 2009

Integration of high-risk human papillomavirus (HPV) types into the host-cell genome disrupts the HPV regulatory E2 protein, resulting in a loss of negative feedback control of viral oncogene expression; this disruption has been considered a critical event in the pathogenesis of cervical neoplasia, and a potential biomarker of progressive disease. However, using serial samples taken from a cohort of young women who were recruited soon after they first had sexual intercourse, we show that disruption of the E2 gene is a common and early event in the natural history of incident cervical HPV infections. The E2 gene was significantly more likely to be disrupted in women who tested positive for HPV18 in their baseline sample than in those who tested positive for HPV16 [26% versus 58%; relative risk, 2.26; 95% confidence interval (CI), 1.38–3.71; χ2, 9.23; 1 degree of freedom (df); P = 0.002]. Among women with an intact E2 gene in their baseline sample, the median time to first detection of E2 disruption was also shorter for those who tested positive for HPV18 than HPV16 (5.7 versus 10.9 months; hazards ratio, 1.93; 95% CI, 0.84–4.44; χ2, 2.49; 1 df; P = 0.11). This tendency for HPV18 to integrate early, coupled with the substantial reduction in viral load in HPV18-positive samples in which E2 is disrupted, may explain why HPV18-associated disease is often reported to be characterized by minor cytologic changes, which underestimate the severity of the underlying histologic abnormality. [Cancer Res 2009;69(9):3828–32]
PI3K Pathway Activation Mediates Resistance to MEK Inhibitors in KRAS Mutant Cancers
Authors: Wee Susan , Jagani Zainab , Xiang Kay Xiaoqin , Loo Alice , Dorsch Marion , Yao Yung-Mae , Sellers William R. , Lengauer Christoph , Stegmeier Frank
Published: April 2009

The RAS pathway is one of the most frequently deregulated pathways in cancer. RAS signals through multiple effector pathways, including the RAF/mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK)/ERK MAPK and phosphatidylinositol 3-kinase (PI3K)-AKT signaling cascades. The oncogenic potential of these effector pathways is illustrated by the frequent occurrence of activating mutations in BRAF and PIK3CA as well as loss-of-function mutations in the tumor suppressor PTEN, a negative regulator of PI3K. Previous studies have found that whereas BRAF mutant cancers are highly sensitive to MEK inhibition, RAS mutant cancers exhibit a more variable response. The molecular mechanisms responsible for this heterogeneous response remain unclear. In this study, we show that PI3K pathway activation strongly influences the sensitivity of RAS mutant cells to MEK inhibitors. Activating mutations in PIK3CA reduce the sensitivity to MEK inhibition, whereas PTEN mutations seem to cause complete resistance. We further show that down-regulation of PIK3CA resensitizes cells with co-occurring KRAS and PIK3CA mutations to MEK inhibition. At the molecular level, the dual inhibition of both pathways seems to be required for complete inhibition of the downstream mammalian target of rapamycin effector pathway and results in the induction of cell death. Finally, we show that whereas inactivation of either the MEK or PI3K pathway leads to partial tumor growth inhibition, targeted inhibition of both pathways is required to achieve tumor stasis. Our study provides molecular insights that help explain the heterogeneous response of KRAS mutant cancers to MEK pathway inhibition and presents a strong rationale for the clinical testing of combination MEK and PI3K targeted therapies. [Cancer Res 2009;69(10):4286–93]
Heregulin Regulates Prolactinoma Gene Expression
Authors: Vlotides George , Cooper Odelia , Chen Yen-Hao , Ren Song-Guang , Greenman Yona , Melmed Shlomo
Published: April 2009

To investigate the role of p185her2/neu/ErbB3 signaling in pituitary tumor function, we examined these receptors in human prolactinomas. Immunofluorescent p185her2/neu was detected in almost all (seven of eight), and ErbB3 expression in a subset (four of eight) of tumors (seven adenomas and one carcinoma). Quantitative PCR also showed abundant ErbB3 mRNA in tumor specimens derived from a rarely encountered prolactin-cell carcinoma. Activation of p185c-neu/ErbB3 signaling with heregulin, the ErbB3 ligand, in rat lacto-somatotroph (GH4C1) tumor cells specifically induced prolactin (PRL) mRNA expression ∼5-fold and PRL secretion ∼4-fold, whereas growth hormone expression was unchanged. Heregulin (6 nmol/L) induced tyrosine phosphorylation and ErbB3 and p185c-neu heterodimerization, with subsequent activation of intracellular ERK and Akt. The Akt signal was specific to ErbB3 activation by heregulin, and was not observed in response to epidermal growth factor activation of epidermal growth factor receptor. Gefitinib, the tyrosine kinase inhibitor, suppressed heregulin-mediated p185c-neu/ErbB3 signaling to PRL. Heregulin induction of PRL was also abrogated by transfecting cells with short interfering RNA directed against ErbB3. Pharmacologic inhibition of heregulin-induced phosphoinositide-3-kinase/Akt (with LY294002) and ERK (with U0126) signaling, as well as short interfering RNA–mediated mitogen-activated protein kinase-1 down-regulation, showed ERK signaling as the primary transducer of heregulin signaling to PRL. These results show ErbB3 expression in human prolactinomas and a novel ErbB3-mediated mechanism for PRL regulation in experimental lactotroph tumors. Targeted inhibition of up-regulated p185c-neu/ErbB3 activity could be useful for the treatment of aggressive prolactinomas resistant to conventional therapy. [Cancer Res 2009;69(10):4209–16]
Enhancing the Antitumor Activity of Adriamycin and Ionizing Radiation
Authors: Sun Wenqing , Kalen Amanda L. , Smith Brian J. , Cullen Joseph J. , Oberley Larry W.
Published: April 2009

Overexpression of manganese superoxide dismutase (MnSOD), when combined with certain chemicals that inhibit peroxide removal, increases cancer cell cytotoxicity. Elevating MnSOD levels in cells enhances the conversion of superoxide (O2•−) to hydrogen peroxide (H2O2), combined with inhibiting the removal of H2O2, further increases H2O2 levels, leading to increased cytotoxicity. We hypothesized that increasing endogenous O2•− production in cells that were pretreated with adenoviral MnSOD (AdMnSOD) plus 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) would lead to an increased level of intracellular H2O2 accumulation and increased cell killing. The cytotoxic effects of Adriamycin or radiation, agents known to produce O2•−, were determined in MDA-MB-231 breast cancer cells pretreated with AdMnSOD plus BCNU both in vitro and in vivo. In vitro, AdMnSOD plus BCNU sensitized cells to the cytotoxicity of Adriamycin or radiation. In vivo, AdMnSOD, BCNU, and Adriamycin or ionizing radiation inhibited tumor growth and prolonged survival. The results suggest that agents that produce O2•− in combination with AdMnSOD plus BCNU may represent a powerful new antitumor regimen against breast cancer. [Cancer Res 2009;69(10):4294–300]
Loss of VHL and Hypoxia Provokes PAX2 Up-Regulation in Clear Cell Renal Cell Carcinoma
Authors: Luu Van-Duc , Boysen Gunther , Struckmann Kirsten , Casagrande Silvia , von Teichman Adriana , Wild Peter J. , Sulser Tullio , Schraml Peter , Moch Holger
Published: April 2009

Purpose: The paired box gene 2, PAX2, encodes for a transcription factor that is up-regulated during nephrogenesis and becomes silenced in mature epithelium of the glomeruli, the proximal, and distal tubules. Reactivation of PAX2 has been frequently observed in clear cell renal cell carcinoma (ccRCC), a tumor type characterized by loss of von Hippel-Lindau (VHL) tumor suppressor function. The regulation of PAX2 expression in ccRCC is unknown.

Experimental Design: We applied reporter gene assays to investigate PAX2 promoter regulation. Furthermore, PAX2 expression was determined in ccRCC cell lines under normoxic and hypoxic condition in a VHL wild-type and mutated background. PAX2 expression was also assessed in 831 human ccRCC and correlated with hypoxia-inducible factor α (HIFα) and clinical parameters.

Results: Here, we show that both loss of VHL protein (pVHL) function and hypoxia leads to strong PAX2 reexpression. Using luciferase reporter gene assays, no induction was obtained in spite of six hypoxia response element motifs identified in the promoter of PAX2. Comprehensive immunohistochemical analyses showed significant correlations between PAX2, HIF1α, and HIF2α—target CCND1 expression patterns in ccRCC patients. Notably, PAX2 expression was highly associated with early-stage, well-differentiated ccRCC and, consequently, better clinical outcome (P < 0.0001 each). Additional analyses indicated that PAX2 repressor WT1 and cancer-linked hypomethylation are not important for transcriptional regulation of PAX2 in ccRCC.

Conclusion: We conclude that in ccRCC, PAX2 reactivation is driven by HIF-dependent mechanisms following pVHL loss.
In vitroandIn vivoCharacterization of MDX-1401 for Therapy of Malignant Lymphoma
Authors: Cardarelli Pina M. , Moldovan-Loomis Maria-Cristina , Preston Ben , Black Amelia , Passmore David , Chen Tseng-Hui , Chen Sharline , Liu Jie , Kuhne Michelle R. , Srinivasan Mohan , Assad Albert , Witte Alison , Graziano Robert F. , King David J.
Published: April 2009

Purpose: This study was undertaken to evaluate the effects of MDX-1401, a nonfucosylated fully human monoclonal antibody that binds to human CD30, and to determine whether it exhibits greater in vitro and in vivo activity than its parental antibody.

Experimental Design: Assays measuring antibody binding to CD30-expressing cells and FcγRIIIa (CD16) transfectants as well as antibody-dependent cellular cytotoxicity (ADCC) were conducted. Antitumor activity was determined using a Karpas-299 systemic model.

Results: The binding of MDX-1401 to CD30 antigen was identical to fucose-containing parental anti-CD30 antibody (MDX-060). In contrast, MDX-1401 showed increased binding affinity to FcγRIIIa-transfected cells resulting in increased effector function. MDX-1401 greatly improved ADCC activity as evidenced by a decrease in half-maximal effective concentration (EC50) and an increase in maximum cell lysis when compared with MDX-060. Increased ADCC activity was observed among a panel of cell lines, including one with very low CD30 antigen expression in which parental antibody failed to induce any detectable ADCC. MDX-1401 activity with all FcγRIIIa polymorphic variants, including less active Phe/Phe158 and Phe/Val158 effector cells, was shown. Furthermore, MDX-1401 was efficacious in inhibiting tumor growth in CD30+ lymphoma xenografts.

Conclusions: The low doses of antibody required for ADCC activity irrespective of donor genotype, the ability to mediate ADCC in target cells expressing low levels of CD30, and increased in vivo efficacy support the development of MDX-1401 for treatment of malignant lymphoma.
Abnormalities in Glucose Uptake and Metabolism in Imatinib-Resistant Human BCR-ABL–Positive Cells
Authors: Kominsky Douglas J. , Klawitter Jelena , Brown Jaimi L. , Boros Laszlo G. , Melo Junia V. , Eckhardt S. Gail , Serkova Natalie J.
Published: April 2009

The development of imatinib resistance has become a significant therapeutic problem in which the etiology seems to be multifactorial and poorly understood. As of today, clinical criteria to predict the development of imatinib resistance in chronic myelogenous leukemia (CML), other than rebound of the myeloproliferation, are under development. However, there is evidence that the control of glucose-substrate flux is an important mechanism of the antiproliferative action of imatinib because imatinib-resistant gastrointestinal stromal KIT-positive tumors reveal highly elevated glucose uptake in radiologic images. We used nuclear magnetic resonance spectroscopy and gas chromatography mass spectrometry to assess 13C glucose uptake and metabolism (glycolysis, TCA cycle, and nucleic acid ribose synthesis) during imatinib treatment in CML cell lines with different sensitivities to imatinib. Our results show that sensitive K562-s and LAMA84-s BCR-ABL–positive cells have decreased glucose uptake, decreased lactate production, and an improved oxidative TCA cycle following imatinib treatment. The resistant K562-r and LAMA84-r cells maintained a highly glycolytic metabolic phenotype with elevated glucose uptake and lactate production. In addition, oxidative synthesis of RNA ribose from 13C-glucose via glucose-6-phosphate dehydrogenase was decreased, and RNA synthesis via the nonoxidative transketolase pathway was increased in imatinib-resistant cells. CML cells which exhibited a (oxidative/nonoxidative) flux ratio for nucleic acid ribose synthesis of >1 were sensitive to imatinib. The resistant K562-r and LAMA84-r exhibited a (oxidative/nonoxidative) flux ratio of <0.7. The changes in glucose uptake and metabolism were accompanied by intracellular translocation of GLUT-1 from the plasma membrane into the intracellular fraction in sensitive cells treated with imatinib, whereas GLUT-1 remained located at the plasma membrane in LAMA84-r and K562-r cells. The total protein load of GLUT-1 was unchanged among treated sensitive and resistant cell lines. In summary, elevated glucose uptake and nonoxidative glycolytic metabolic phenotype can be used as sensitive markers for early detection of imatinib resistance in BCR-ABL–positive cells.
Cancer Prevention and Treatment with Resveratrol: From Rodent Studies to Clinical Trials
Authors: Bishayee Anupam
Published: April 2009

Resveratrol (3,4′,5-trihydroxy-trans-stilbene) is a dietary polyphenol derived from grapes, berries, peanuts, and other plant sources. During the last decade, resveratrol has been shown to possess a fascinating spectrum of pharmacologic properties. Multiple biochemical and molecular actions seem to contribute to resveratrol effects against precancerous or cancer cells. Resveratrol affects all three discrete stages of carcinogenesis (initiation, promotion, and progression) by modulating signal transduction pathways that control cell division and growth, apoptosis, inflammation, angiogenesis, and metastasis. The anticancer property of resveratrol has been supported by its ability to inhibit proliferation of a wide variety of human tumor cells in vitro. These in vitro data have led to numerous preclinical animal studies to evaluate the potential of this drug for cancer chemoprevention and chemotherapy. This review provides concise, comprehensive data from preclinical in vivo studies in various rodent models of human cancers, highlighting the related mechanisms of action. Bioavailability, pharmacokinetic, and potential toxicity studies of resveratrol in humans and ongoing interventional clinical trials are also presented. The conclusion describes directions for future resveratrol research to establish its activity and utility as a human cancer preventive and therapeutic drug.
Methyl-Selenium Compounds Inhibit Prostate Carcinogenesis in the Transgenic Adenocarcinoma of Mouse Prostate Model with Survival Benefit
Authors: Wang Lei , Bonorden Melissa J.L. , Li Guang-xun , Lee Hyo-Jeong , Hu Hongbo , Zhang Yong , Liao Joshua D. , Cleary Margot P. , Lü Junxuan
Published: April 2009

Chemoprevention of prostate cancer by second-generation selenium compounds in reference to selenomethionine holds strong promise to deal with the disease at the root. Here we used the transgenic adenocarcinoma mouse prostate (TRAMP) model to establish the efficacy of methylseleninic acid (MSeA) and methylselenocysteine (MSeC) against prostate carcinogenesis and to characterize potential mechanisms. Eight-week-old male TRAMP mice (C57B/6 background) were given a daily oral dose of water, MSeA, or MSeC at 3 mg Se/kg body weight and were euthanized at either 18 or 26 weeks of age. By 18 weeks of age, the genitourinary tract and dorsolateral prostate weights for the MSeA- and MSeC-treated groups were lower than for the control (P < 0.01). At 26 weeks, 4 of 10 control mice had genitourinary weight >2 g, and only 1 of 10 in each of the Se groups did. The efficacy was accompanied by delayed lesion progression, increased apoptosis, and decreased proliferation without appreciable changes of T-antigen expression in the dorsolateral prostate of Se-treated mice and decreased serum insulin-like growth factor I when compared with control mice. In another experiment, giving MSeA to TRAMP mice from 10 or 16 weeks of age increased their survival to 50 weeks of age, and delayed the death due to synaptophysin-positive neuroendocrine carcinomas and synaptophysin-negative prostate lesions and seminal vesicle hypertrophy. Wild-type mice receiving MSeA from 10 weeks did not exhibit decreased body weight or genitourinary weight or increased serum alanine aminotransferase compared with the control mice. Therefore, these selenium compounds may effectively inhibit this model of prostate cancer carcinogenesis.
Glutathione S-Transferase Polymorphisms and Risk of Second Primary Malignancy after Index Squamous Cell Carcinoma of the Head and Neck
Authors: Zafereo Mark E. , Sturgis Erich M. , Aleem Sal , Chaung Katrina , Wei Qingyi , Li Guojun
Published: April 2009

Glutathione S-transferases (GST) detoxify carcinogens in tobacco smoke, which plays a major role in development of not only squamous cell carcinoma of the head and neck (SCCHN) but also second primary malignancy (SPM) after index SCCHN.

We hypothesized that GSTM1 null, GSTT1 null, GSTP1 Ile105Val, and GSTP1 Ala114Val polymorphisms would individually and, more likely, collectively show an association with risk of SPM after index SCCHN. One thousand three hundred seventy-six incident SCCHN patients were prospectively recruited between May 1996 and December 2006, genotyped, and followed for SPM development.

One hundred ten patients (8%) developed SPM: 43 (39%) second SCCHN, 38 (35%) other tobacco-associated sites, and 29 (26%) other non–tobacco-associated sites. Patients with GSTP1 Ile105Val polymorphism had a statistically significant association with risk of SPM development (adjusted hazard ratio, 1.7; 95% confidence interval, 1.1-2.5). However, no statistically significant associations were observed with GSTM1, GSTT1, or GSTP1 Ala114Val polymorphisms. After combining risk genotypes for all four polymorphisms, rates of SPM development with 0 to 1, 2, 3, and 4 risk genotypes were 6.4%, 8.4%, 10.9%, and 15.1%, respectively, and a stepwise increase in SPM risk was observed with increasing number of risk genotypes (P = 0.004 for trend). Patients with 3 to 4 risk genotypes had a 1.7-fold increased risk for SPM compared with patients with 0 to 2 risk genotypes (hazard ratio, 1.70; 95% confidence interval, 1.2-2.5).

This large prospective cohort study supports a modestly increased risk of SPM after index SCCHN with GSTP1 Ile105Val polymorphism and an even greater risk of SPM with multiple combined GST risk genotypes.
Constitutive Short Telomere Length of Chromosome 17p and 12q but not 11q and 2p Is Associated with an Increased Risk for Esophageal Cancer
Authors: Xing Jinliang , Ajani Jaffer A. , Chen Meng , Izzo Julie , Lin Jie , Chen Zhinan , Gu Jian , Wu Xifeng
Published: April 2009

Shortened telomere length may cause chromosomal instability in Barrett's esophagus and thus promote tumorigenesis. However, whether short telomere length in all chromosomes or just some of them is associated with increased esophageal cancer (EC) risk is largely unknown. To address this question, we examined the overall and chromosome-specific telomere lengths of 17p, 12q, 2p, and 11q and assessed their associations with EC risk. In a case-control study with 94 EC cases and 94 matched controls, the overall telomere length and the chromosome-specific telomere lengths of 17p, 12q, 2p, and 11q in peripheral blood lymphocytes were determined by a real-time PCR and a modified single telomere length analysis assay, respectively. Multivariate logistic regression analysis was used to assess the association between telomere length and EC risk. Compared with controls, EC patients had significantly shorter overall telomere lengths (P = 0.004) and chromosome-specific telomere lengths of 17p (P = 0.003) and 12q (P = 0.006) but not of 11q (P = 0.632) and 2p (P = 0.972). Furthermore, the multivariate logistic regression analysis showed that the short overall telomere length and chromosome-specific telomere lengths of 17p and 12q were associated with a dose-dependent increase in EC risk. Our study provides the first epidemiologic evidence that short telomere length of 17p and 12q plays an important role in esophageal carcinogenesis, suggesting that short telomere length of specific chromosomes is associated with the etiology of different cancer types.
Objective Detection and Delineation of Oral Neoplasia Using Autofluorescence Imaging
Authors: Roblyer Darren , Kurachi Cristina , Stepanek Vanda , Williams Michelle D. , El-Naggar Adel K. , Lee J. Jack , Gillenwater Ann M. , Richards-Kortum Rebecca
Published: April 2009

Although the oral cavity is easily accessible to inspection, patients with oral cancer most often present at a late stage, leading to high morbidity and mortality. Autofluorescence imaging has emerged as a promising technology to aid clinicians in screening for oral neoplasia and as an aid to resection, but current approaches rely on subjective interpretation. We present a new method to objectively delineate neoplastic oral mucosa using autofluorescence imaging.

Autofluorescence images were obtained from 56 patients with oral lesions and 11 normal volunteers. From these images, 276 measurements from 159 unique regions of interest (ROI) sites corresponding to normal and confirmed neoplastic areas were identified. Data from ROIs in the first 46 subjects were used to develop a simple classification algorithm based on the ratio of red-to-green fluorescence; performance of this algorithm was then validated using data from the ROIs in the last 21 subjects. This algorithm was applied to patient images to create visual disease probability maps across the field of view. Histologic sections of resected tissue were used to validate the disease probability maps.

The best discrimination between neoplastic and nonneoplastic areas was obtained at 405 nm excitation; normal tissue could be discriminated from dysplasia and invasive cancer with a 95.9% sensitivity and 96.2% specificity in the training set, and with a 100% sensitivity and 91.4% specificity in the validation set. Disease probability maps qualitatively agreed with both clinical impression and histology. Autofluorescence imaging coupled with objective image analysis provided a sensitive and noninvasive tool for the detection of oral neoplasia.
Short-term Prophylactic Tamoxifen Reduces the Incidence of Antiestrogen-Resistant/Estrogen Receptor–Positive/Progesterone Receptor–Negative Mammary Tumors
Authors: Rose-Hellekant Teresa A. , Skildum Andrew J. , Zhdankin Olga , Greene Amy L. , Regal Ronald R. , Kundel Katherine D. , Kundel Donald W.
Published: April 2009

Although many estrogen receptor–positive (ER+) breast cancers are effectively treated with selective estrogen receptor modulators and down-regulators (SERM/SERD), some are highly resistant. Resistance is more likely if primary cancers are devoid of progesterone receptors (PR−) or have high levels of growth factor activity. In this study, a transgenic mouse line that expresses transforming growth factor-α (NRL-TGFα mice) and that develops ER+/PR− mammary tumors was used to assess the possible effects of (a) therapeutic delivery of the SERM, tamoxifen, or SERD, ICI I82,780 (ICI), on the growth of established tumors and (b) short-term prophylactic tamoxifen administration on the initial development of new mammary tumors. To determine the therapeutic effects of tamoxifen and ICI on the growth of established tumors, mice were exposed to 3 weeks of drug treatment. Neither drug influenced tumor growth or glandular pathology. To determine if early prophylactic tamoxifen could alter tumorigenesis, a 60-day tamoxifen treatment was initiated in 8-week-old mice. Compared with placebo-treated mice, tamoxifen reduced tumor incidence by 50% and significantly decreased the degree of mammary hyperplasia. Prophylactic tamoxifen also significantly extended the life span of tumor-free mice. These data show that in this mouse model, established ER+/PR− mammary tumors are resistant to SERM/SERD treatment but the development of new mammary tumors can be prevented by an early course of tamoxifen. This study validates the utility of NRL-TGFα mice for (a) identifying candidate biomarkers of efficacious tamoxifen chemoprevention and (b) modeling the evolution of tamoxifen resistance.
Nonsteroidal Anti-inflammatory Drug-Activated Gene-1 Expression Inhibits Urethane-Induced Pulmonary Tumorigenesis in Transgenic Mice
Authors: Cekanova Maria , Lee Seong-Ho , Donnell Robert L. , Sukhthankar Mugdha , Eling Thomas E. , Fischer Susan M. , Baek Seung Joon
Published: April 2009

The expression of nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1) inhibits gastrointestinal tumorigenesis in NAG-1 transgenic mice (C57/BL6 background). In the present study, we investigated whether the NAG-1 protein would alter urethane-induced pulmonary lesions in NAG-1 transgenic mice on an FVB background (NAG-1Tg+/FVB). NAG-1Tg+/FVB mice had both decreased number and size of urethane-induced tumors, compared with control littermates (NAG-1Tg+/FVB = 16 ± 4 per mouse versus control = 20 ± 7 per mouse, P < 0.05). Urethane-induced pulmonary adenomas and adenocarcinomas were observed in control mice; however, only pulmonary adenomas were observed in NAG-1Tg+/FVB mice. Urethane-induced tumors from control littermates and NAG-1Tg+/FVB mice highly expressed proteins in the arachidonic acid pathway (cyclooxygenases 1/2, prostaglandin E synthase, and prostaglandin E2 receptor) and highly activated several kinases (phospho-Raf-1 and phosphorylated extracellular signal-regulated kinase 1/2). However, only urethane-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation was decreased in NAG-1Tg+/FVB mice. Furthermore, significantly increased apoptosis in tumors of NAG-1Tg+/FVB mice compared with control mice was observed as assessed by caspase-3/7 activity. In addition, fewer inflammatory cells were observed in the lung tissue isolated from urethane-treated NAG-1Tg+/FVB mice compared with control mice. These results paralleled in vitro assays using human A549 pulmonary carcinoma cells. Less phosphorylated p38 MAPK was observed in cells overexpressing NAG-1 compared with control cells. Overall, our study revealed for the first time that the NAG-1 protein inhibits urethane-induced tumor formation, probably mediated by the p38 MAPK pathway, and is a possible new target for lung cancer chemoprevention.
Progress in Genome-Wide Association Studies of Human Height
Authors: Hirschhorn Joel N. , Lettre Guillaume
Published: April 2009

Human height (stature) is a strongly genetic trait, with up to 90% of the variation in height within a population determined by a combination of multiple inherited factors. Recent advances in genetics and genomics now permit comprehensive genome-wide surveys of common genetic variations in those variants that are associated with stature. The first such studies have borne fruit, identifying over 40 genetic loci that can be reproducibly shown to have an influence on adult height. These unbiased searches throughout the genome identified several loci that also harbour rare mutations responsible for more severe alterations in height or skeletal growth. Although the predictive value of the common variants thus far discovered remains low, the identification of these loci has led to new insights into the biology of human growth, and may help identify genes that underlie previously uncharacterized syndromes of abnormal skeletal growth.
The Candidate Gene Approach to the Diagnosis of Monogenic Disorders
Authors: Dattani Mehul T.
Published: April 2009

The genetic basis of many human diseases has been elucidated by the candidate gene approach. Mouse models of human disease, whether naturally occurring or derived by transgenesis, have enabled a number of disease genes to be identified. Such approaches have formed the basis of the identification of candidate genes implicated in hypothalamo-pituitary development. Nevertheless, significant differences exist between murine and human models of disease. Chromosomal deletions, duplications and translocations that are visible microscopically are also informative and have allowed the discovery of disease genes such as SOX2 and SOX3. In addition, genome-wide mapping techniques using microsatellites have led to the identification of genes such as GPR54, mutations of which are associated with hypogonadotrophic hypogonadism. Newer techniques, such as array comparative genomic hybridization (CGH), have enabled the detection of submicroscopic chromosomal imbalances. These, in turn, can also lead to the identification of disease genes, as has been demonstrated by the discovery of mutations in CHD7 in some patients with the CHARGE syndrome. Most significant, however, is the recent optimization of high-density whole-genome single-nucleotide polymorphism arrays, which provides the potential to identify many disease genes and may revolutionize the field of human genetics.
Epigenetics and Imprinted Genes: Insights from the Imprinted Gnas Locus
Authors: Kelsey Gavin
Published: April 2009

The hallmarks of epigenetics – the memory of defining earlier developmental events and the distinction of active and inactive genes – are exemplified by imprinted genes. In this article, I shall consider the imprinted Gnas locus in some detail. Gnas encodes the stimulatory G-protein subunit, Gsα, an essential intermediate between receptor coupling and cyclic adenosine monophosphate generation. It provides an excellent illustration of the pleiotropic effects of imprinted genes, particularly on skeletal growth and metabolism, and is a powerful example of the conflicting effects of imprinted genes with opposing patterns of imprinting. I shall describe the effects of Gsα deficiency in humans and the knowledge gained from genetic manipulation in the mouse. Finally, given the pervasive effects of imprinted genes, I shall discuss the likelihood that epigenetic deregulation, for example of imprinted genes, could contribute to the developmental programming of chronic adult diseases.