bluuebunny/crossref_metadata_2025 huggingface dataset:
Actinometry Measurement of Dissociation Degrees of Nitrogen and Oxygen in N2–O2Microwave Discharge Plasma
Authors: Ichikawa Yosuke , Sakamoto Takeshi , Nezu Atsushi , Matsuura Haruaki , Akatsuka Hiroshi
Published: October 2010

The dissociation degrees of N2and O2are examined in a nitrogen–oxygen mixed microwave discharge plasma in a cylindrical quartz tube of 26 mm inner diameter with a discharge pressure of 0.5–1.0 Torr and a microwave power of 600 W by the actinometry method. We measured the electron temperature and density with a Langmuir double probe, while the vibrational and rotational temperatures of the first and second positive bands of N2were measured by optical emission spectroscopy. Even when the line intensity of atomic nitrogen was weak and partly coincided with the high-intensity band spectrum of the first positive system due to its small dissociation degree, the actinometry method was found to be feasible when the first positive band spectrum, calculated as a function of the rotational and vibrational temperatures, was subtracted from that observed experimentally. It was found that the dissociation degrees of both N2and O2increase with the molar ratio of nitrogen in the mixed N2–O2discharge gas for the same total discharge pressure. The experimental results are discussed by comparison with a simple numerical model based on chemical kinetics in the plasma. It was found that the dissociation of oxygen molecules is enhanced by the collision with excited nitrogen molecules, particularly those with metastable states, whereas that of nitrogen is suppressed by an admixture of oxygen molecules due to the chemical quenching processes of nitrogen atoms.
Dynamics of Radical Cation of Poly(4-hydroxystyrene)-Based Chemically Amplified Resists for Extreme-Ultraviolet and Electron Beam Lithographies
Authors: Okamoto Kazumasa , Tanaka Masafumi , Kozawa Takahiro , Tagawa Seiichi
Published: October 2010

The elucidation of the reaction mechanism after the irradiation of ionizing radiation onto resist polymers is an important issue for extreme-ultraviolet and electron beam chemically amplified resist. The dynamics of radical cations of polymers is essential for elucidating the mechanism of acid formation (deprotonation) in resists. The transient absorption of poly(4-methoxystyrene) (PMOS) was observed in 1,2-dichloroethane and p-dioxane solutions by pulse radiolysis. In the near-infrared region, a characteristic charge resonance band, which represents the π–π interaction between the two benzene rings of the intramolecular PMOS dimer radical cation [(Ph–OCH3)2
+·], was observed. In the presence of halogenated hydrocarbon molecules acting as electron scavengers, the yield of (Ph–OCH3)2
+· was enhanced by the formation of an ion/charge transfer complex. Density functional theory (DFT) calculations were also performed to investigate the electronic state of (Ph–OCH3)2
+·.
Fabrication of Hierarchically Micro- and Nano-structured Mold Surfaces Using Laser Ablation for Mass Production of Superhydrophobic Surfaces
Authors: Noh Jiwhan , Lee Jae-Hoon , Na Suckjoo , Lim Hyuneui , Jung Dae-Hwan
Published: October 2010

Many studies have examined the formation of surfaces with mixed patterns of micro- and nano-sized lotus leaves that have hydrophobic properties. In this study, micro- and nano-shapes such as lotus leaves were fabricated on a metal mold surface using laser ablation and ripple formation. A microstructure on the mold surface was replicated onto poly(dimethylsiloxane) (PDMS) using the polymer casting method to manufacture low-cost hydrophobic surfaces. A PDMS surface with micro- and nano-structures that were the inverse image of a lotus leaf showed hydrophobic characteristics (water contact angle: 157°). From these results, we deduced that portions of the microstructures were wet and that air gaps existed between the microstructures and the water drops. In this paper we suggest the possibility of the mass production of hydrophobic plastic surfaces and the development of a methodology for the hydrophobic texturing of various polymer surfaces, using the polymer casting method with laser-processed molds.
Fabrication of Random Microspikes on Mold Metal by Ultrashort Laser Ablation for Hydrophilic Surface
Authors: Noh Jiwhan , Lee Jae-Hoon , Lee Sang-Yup , Na Suckjoo
Published: October 2010

Microsized spikes, which are smaller than laser focused spots, were fabricated by laser ablation in this study. The hydrophilic property of the surface with the microsize spikes was tested. Microspikes of 20 µm size were formed using a 50 µm laser focused spot size by optimizing the laser process parameters. Water drops were placed on the microspikes to measure the contact angle, which was 10° while that of the surface without microspikes was 80°. This shows that a surface can be processed to be hydrophilic with laser ablation, without applying hydrophilic chemical coating. Because the base material in this study was a mold metal, injection molding products with functional surfaces were produced economically by forming micropatterns on their surfaces. The economically micropatterns produced in this study will be applicable for controlling the friction of fluids on surfaces, as well as for producing hydrophilic surfaces.
Nanoscale Selective Plasma Etching of Ultrathin HfO2Layers on GaAs for Advanced Complementary Metal–Oxide–Semiconductor Devices
Authors: Anguita Jose , Benedicto Marcos , Alvaro Raquel , Galiana Beatriz , Tejedor Paloma
Published: October 2010

We present a reliable dry-etch process for patterning deep-submicron structures in ultrathin (16 nm) HfO2layers deposited on GaAs substrates. Plasma chemistries based on BCl3/O2and SF6/Ar have been investigated using an inductively-coupled plasma reactive ion etch (ICP-RIE) reactor. The process reliability has been examined in terms of etch rate selectivity, etch time control, anisotropy, and surface roughness of the underlying GaAs substrate for potential application to gate nanopatterning in next-generation field-effect transistor fabrication. We show that a SF6/Ar plasma process provides excellent prospects as a nanopatterning method for subsequent re-growth of GaAs in novel device architectures.
X-ray Microfocusing by Combination of Grazing-Incidence Spherical-Concave Mirrors
Authors: Suzuki Yoshio , Takeuchi Akihisa
Published: October 2010

An optical system for X-ray microfocusing that consists of a pair of spherical mirrors has been developed. In grazing incidence optics with spherical (cylindrical) mirrors, focused beam size is generally restricted by spherical aberration. However, the spherical aberration can be corrected by the sequential reflection of two spherical mirrors, and submicrometer beam size can be obtained by total-reflection mirror optics with spherical concave mirrors. A sub-200 nm focusing has been achieved at an X-ray energy of 8 keV.
Sputter Deposition of High-Mobility Sn1-xTaxO2 Films on Anatase-TiO2-Coated Glass
Authors: Yamada Naoomi , Nakao Shoichiro , Hitosugi Taro , Hasegawa Tetsuya
Published: October 2010

Ta-doped SnO2 (Sn0.99Ta0.01O2; TTO) films were successfully sputter deposited with a high mobility of 49 cm2 V-1 s-1 on glass coated with anatase-TiO2 seed layers. The high mobility enabled the achievement of a film resistivity of 5.9 ×10-4 Ω cm. This value of resistivity is one of the lowest ever reported for SnO2-based polycrystalline films without enhanced free-carrier absorption. An optimized film with a sheet resistance of 14 Ω/□ exhibited a transmittance of more than 70%, even in the near-infrared region. These results demonstrate that sputtered TTO films obtained by the TiO2 seed layer method are suitable for use in transparent electrodes in solar cells.
Integrating streamer and ocean-bottom seismic data for sub-basalt imaging on the Atlantic Margin
Authors: Lau K. W. Helen , White R. S. , Christie P. A. F.
Published: October 2010

Stacked basalt flows cover much of the NW European continental margin, including potentially prospective sediments of the Faroes shelf. Such flows attenuate seismic energy, hindering sub-basalt structural imaging which is critical for both exploration and tectonic studies. Low-frequency, long-offset reflection surveys have yielded improved images below top basalt, while coincident ocean-bottom seismometer (OBS) data have mapped low velocity zones (LVZ) from tomographic inversion. Image correlation in a common depth domain is challenged by low spatial resolution of the tomographic image, the absence of turning rays in a LVZ and the lack of wide-angle arrivals in the reflection data. We integrate densely sampled reflection data with deep velocities from OBS data to give a common velocity model in a new, iterated, pre-stack depth-migration workflow using complementary constraints from the two datasets. The matched velocity model and depth image enable interpretation of (i) a uniform flood basalt sequence, 2–4 km thick beneath the Fugloy Ridge, with velocities correlating with those in the Lopra 1/1A borehole and (ii) a sub-basalt LVZ with chaotic reflections corresponding to sill-intruded, probably syn-rift sediments. Such a workflow could be extended to targets beneath high velocity salt or basalt and could provide constraints for 3D datasets.

Supplementary material:
Overlays of modelled arrival times on all the OBS gathers, similar to Figure 4, can be found at
http://www.geolsoc.org.uk/SUP18433
Adrenal glands are essential for activation of glucogenesis during undernutrition in fetal sheep near term
Authors: Fowden A. L. , Forhead A. J.
Published: October 2010

In adults, the adrenal glands are essential for the metabolic response to stress, but little is known about their role in fetal metabolism. This study examined the effects of adrenalectomizing fetal sheep on glucose and oxygen metabolism in utero in fed conditions and after maternal fasting for 48 h near term. Fetal adrenalectomy (AX) had little effect on the rates of glucose and oxygen metabolism by the fetus or uteroplacental tissues in fed conditions. Endogenous glucose production was negligible in both AX and intact, sham-operated fetuses in fed conditions. Maternal fasting reduced fetal glucose levels and umbilical glucose uptake in both groups of fetuses to a similar extent but activated glucose production only in the intact fetuses. The lack of fasting-induced glucogenesis in AX fetuses was accompanied by falls in fetal glucose ultilization and oxygen consumption not seen in intact controls. The circulating concentrations of cortisol and total catecholamines, and the hepatic glycogen content and activities of key gluconeogenic enzymes, were also less in AX than intact fetuses in fasted animals. Insulin concentrations were also lower in AX than intact fetuses in both nutritional states. Maternal glucose utilization and its distribution between the fetal, uteroplacental, and nonuterine maternal tissues were unaffected by fetal AX in both nutritional states. Ovine fetal adrenal glands, therefore, have little effect on basal rates of fetal glucose and oxygen metabolism but are essential for activating fetal glucogenesis in response to maternal fasting. They may also be involved in regulating insulin sensitivity in utero.
Improved glucose control and reduced body fat mass in free fatty acid receptor 2-deficient mice fed a high-fat diet
Authors: Bjursell Mikael , Admyre Therése , Göransson Melker , Marley Anna E. , Smith David M. , Oscarsson Jan , Bohlooly-Y Mohammad
Published: October 2010

Free fatty acid receptor 2 (Ffar2), also known as GPR43, is activated by short-chain fatty acids (SCFA) and expressed in intestine, adipocytes, and immune cells, suggesting involvement in lipid and immune regulation. In the present study, Ffar2-deficient mice ( Ffar2-KO) were given a high-fat diet (HFD) or chow diet and studied with respect to lipid and energy metabolism. On a HFD, Ffar2-KO mice had lower body fat mass and increased lean body mass. The changed body composition was accompanied by improved glucose control and lower HOMA index, indicating improved insulin sensitivity in Ffar2-KO mice. Moreover, the Ffar2-KO mice had higher energy expenditure accompanied by higher core body temperature and increased food intake. The liver weight and content of triglycerides as well as plasma levels of cholesterol were lower in the Ffar2-KO mice fed a HFD. A histological examination unveiled decreased lipid interspersed in brown adipose tissue of the Ffar2-KO mice. Interestingly, no significant differences in white adipose tissue (WAT) cell size were observed, but significantly lower macrophage content was detected in WAT from HFD-fed Ffar2-KO compared with wild-type mice. In conclusion, Ffar2 deficiency protects from HFD-induced obesity and dyslipidemia at least partly via increased energy expenditure.
Proteomics analysis reveals diabetic kidney as a ketogenic organ in type 2 diabetes
Authors: Zhang Dongjuan , Yang Hang , Kong Xiaomu , Wang Kang , Mao Xuan , Yan Xianzhong , Wang Yuan , Liu Siqi , Zhang Xiaoyan , Li Jing , Chen Lihong , Wu Jing , Wei Mingfen , Yang Jichun , Guan Youfei
Published: October 2010

Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. To date, the molecular mechanisms of DN remain largely unclear. The present study aimed to identify and characterize novel proteins involved in the development of DN by a proteomic approach. Proteomic analysis revealed that 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase 2 (HMGCS2), the key enzyme in ketogenesis, was increased fourfold in the kidneys of type 2 diabetic db/db mice. Consistently, the activity of HMGCS2 in kidneys and 24-h urinary excretion of the ketone body β-hydroxybutyrate (β-HB) were significantly increased in db/db mice. Immunohistochemistry, immunofluorescence, and real-time PCR studies further demonstrated that HMGCS2 was highly expressed in renal glomeruli of db/db mice, with weak expression in the kidneys of control mice. Because filtered ketone bodies are mainly reabsorbed in the proximal tubules, we used RPTC cells, a rat proximal tubule cell line, to examine the effect of the increased level of ketone bodies. Treating cultured RPTC cells with 1 mM β-HB significantly induced transforming growth factor-β1 expression, with a marked increase in collagen I expression. β-HB treatment also resulted in a marked increase in vimentin protein expression and a significant reduction in E-cadherin protein levels, suggesting an enhanced epithelial-to-mesenchymal transition in RPTCs. Collectively, these findings demonstrate that diabetic kidneys exhibit excess ketogenic activity resulting from increased HMGCS2 expression. Enhanced ketone body production in the diabetic kidney may represent a novel mechanism involved in the pathogenesis of DN.
Exercise training augments the peripheral insulin-sensitizing effects of pioglitazone in HIV-infected adults with insulin resistance and central adiposity
Authors: Yarasheski Kevin E. , Cade W. Todd , Overton E. Turner , Mondy Kristin E. , Hubert Sara , Laciny Erin , Bopp Coco , Lassa-Claxton Sherry , Reeds Dominic N.
Published: October 2010

The prevalence and incidence of insulin resistance and type 2 diabetes mellitus (DM) are higher in people treated for human immunodeficiency virus-1 (HIV) infection than in the general population. Identifying safe and effective interventions is a high priority. We evaluated whether the peroxisome proliferator-activated receptor-γ agonist pioglitazone with exercise training improves central and peripheral insulin sensitivity more than pioglitazone alone in HIV-infected adults with insulin resistance and central adiposity. Forty-four HIV-infected adults with baseline insulin resistance and central adiposity were randomly assigned to 4 mo of pioglitazone (30 mg/day) with or without supervised, progressive aerobic, and resistance exercise training (1.5–2 h/day, 3 days/wk). The hyperinsulinemic euglycemic clamp was used to evaluate alterations in central and peripheral insulin sensitivity. Thirty-nine participants completed the study. Hepatic insulin sensitivity improved similarly in both groups. Exercise training augmented the beneficial effects of pioglitazone on peripheral insulin sensitivity. Greater improvements in peripheral insulin sensitivity were associated with reductions in total body and limb adipose content rather than increases in limb adiposity or pioglitazone-induced increases in adiponectin concentration. We conclude that supplementing pioglitazone with increased physical activity improved insulin sensitivity more effectively than pioglitazone alone in HIV-infected adults with insulin resistance and central adiposity. Pioglitazone alone did not significantly increase limb adipose content. Potential cardiovascular benefits of these interventions in HIV need investigation.
Free fatty acids in the presence of high glucose amplify monocyte inflammation via Toll-like receptors
Authors: Dasu Mohan R. , Jialal Ishwarlal
Published: October 2010

Type 2 diabetes (T2DM) is characterized by hyperglycemia, dyslipidemia, and increased inflammation. Previously, we showed that high glucose (HG) induces Toll-like receptor (TLR) expression, activity, and inflammation via NF-κB followed by cytokine release in vitro and in vivo. Here, we determined how HG-induced inflammation is affected by free fatty acids (FFA) in human monocytes. THP-1 monocytic cells, CD14+human monocytes, and transiently transfected HEK293 cells were exposed to various FFA (0–500 μM) and glucose (5–20 mM) for evaluation of TLR2, TLR4, NF-κB, IL-1β, monocyte chemoattractant protein-1 (MCP-1), and superoxide release. In THP-1 cells, palmitate increased cellular TLR2 and TLR4 expression, generated reactive oxygen species (ROS), and increased NF-κB activity, IL-1β, and MCP-1 release in a dose- and time-dependent manner. Similar data were observed with stearate and FFA mixture but not with oleate. Conversely, NADPH oxidase inhibitor treatment repressed glucose- and palmitate-stimulated ROS generation and NF-κB activity and decreased IL-1β and MCP-1 expression. Silencing TLR2, TLR4, and p47phox with small inhibitory RNAs (siRNAs) significantly reduced superoxide release, NF-κB activity, IL-1β, and MCP-1 secretion in HG and palmitate-treated THP-1 cells. Moreover, data from transient transfection experiments suggest that TLR6 is required for TLR2 and MD2 for TLR4 to augment inflammation in FFA- and glucose-exposed cells. These findings were confirmed with human monocytes. We conclude that FFA exacerbates HG-induced TLR expression and activity in monocytic cells with excess superoxide release, enhanced NF-κB activity, and induced proinflammatory factor release.
Regulation of sFlt-1 and VEGF secretion by adenosine under hypoxic conditions in rat placental villous explants
Authors: George Eric M. , Cockrell Kathy , Adair Thomas H. , Granger Joey P.
Published: October 2010

The role of adenosine in the regulation of cardiovascular function has long been acknowledged, but only recently has its importance in angiogenesis been appreciated, most notably, through its direct regulation of the proangiogenic growth factor, VEGF. Recent work has established that proangiogenic and antiangiogenic factors, specifically VEGF and and the soluble VEGF receptor fms-like tyrosine kinase-1 (sFlt-1), are directly influenced by hypoxia in placental ischemia. While adenosine has been reported to be an important regulator of VEGF in vascular tissue, the importance of adenosine in regulating VEGF and sFlt-1 in placental tissue is unclear. Here, we have investigated the role of adenosine in the secretion of VEGF and the antiangiogenic protein sFlt-1 in placental villous explants. Under normoxic conditions (6% oxygen), the nonspecific adenosine receptor antagonist, 8-sulphophenyltheophylline (8-SPT) had no effect on either VEGF ( P = 0.38) or sFlt-1 ( P = 0.56) secretion. However, under hypoxic conditions (1% oxygen), 8-SPT attenuated the increase in the secretion of both VEGF and sFlt-1 ( P < 0.05 and P < 0.005, respectively). Exogenous and the adenosine transporter inhibitor dipyridamole (which increases extracellular levels of adenosine) showed differential effects under normoxic conditions: sFlt-1 levels in media increased significantly ( P < 0.05), whereas VEGF was unaffected ( P = 0.67 and P = 0.19, respectively). These data indicate that extracellular adenosine can regulate VEGF and sFlt-1 secretion in the hypoxic placenta and could, therefore, control the balance of these competing angiogenic factors in diseases characterized by placental ischemia.
Renal denervation modulates angiotensin receptor expression in the renal cortex of rabbits with chronic heart failure
Authors: Clayton Sarah C. , Haack Karla K. V. , Zucker Irving H.
Published: October 2010

Excessive sympathetic drive is a hallmark of chronic heart failure (HF). Disease progression can be correlated with plasma norepinephrine concentration. Renal function is also correlated with disease progression and prognosis. Because both the renal nerves and renin-angiotensin II system are activated in chronic HF we hypothesized that excessive renal sympathetic nerve activity decreases renal blood flow in HF and is associated with changes in angiotensin II type 1 receptor (AT1R) and angiotensin II type 2 receptor (AT2R) expression. The present study was carried out in conscious, chronically instrumented rabbits with pacing-induced HF. We found that rabbits with HF showed a decrease in mean renal blood flow (19.8 ± 1.6 in HF vs. 32.0 ± 2.5 ml/min from prepace levels; P < 0.05) and an increase in renal vascular resistance (3.26 ± 0.29 in HF vs. 2.21 ± 0.13 mmHg·ml−1·min in prepace normal rabbits; P < 0.05) while the blood flow and resistance was not changed in HF rabbits with the surgical renal denervation. Renal AT1R expression was increased by ∼67% and AT2R expression was decreased by ∼87% in rabbits with HF; however, kidneys from denervated rabbits with HF showed a near normalization in the expression of these receptors. These results suggest renal sympathetic nerve activity elicits a detrimental effect on renal blood flow and may be associated with alterations in the expression of angiotensin II receptors.
A Within Trial Measure of the Stop Signal Reaction Time in a Head-Unrestrained Oculomotor Countermanding Task
Authors: Goonetilleke Samanthi C. , Doherty Timothy J. , Corneil Brian D.
Published: October 2010

The countermanding (or stop-signal) task, which requires the cancellation of an impending response on the infrequent presentation of a stop signal, enables study of the contextual control of movement generation and suppression. Here we present a novel and empirical measure of the time needed to cancel an impending gaze shift by recording neck muscle activity during a head-unrestrained oculomotor countermanding paradigm. On a subset of stop signal trials, subjects generated small head movements toward a target even though gaze remained stable due to a compensatory vestibular-ocular reflex. On such trials, we observed a burst of antagonist neck muscle activity during the small head-only error. Such antagonist neck muscle activity served as an active braking pulse as its magnitude scaled with the kinematics of the head-only error. This activity was selective for trials in which the head was arrested in mid-flight and did not appear on trials without a stop signal, on noncancelled stop signal trials when the gaze shift was completed, or on stop signal trials without head motion. Importantly, the timing of this antagonist activity related best to the onset of the stop signal (lagging it by ∼180 ms), and strongly correlated with behavioral estimates of the time needed to cancel a movement (the stop signal reaction time). These results are consistent with the notion that such selective antagonist neck muscle activity arises as a peripheral expression of the oculomotor stop process that successfully cancelled the gaze shift. Studying movement cancellation within nested systems like the head-unrestrained gaze shifting system offers a unique opportunity for investigating underlying neural mechanisms as the overall goal (i.e., to cancel a gaze shift) can be achieved despite motion of other components; on such individual trials, the oculomotor stop process is expressed as an active braking pulse.
Does Gender Impact Intensity of Care Provided to Older Medical Intensive Care Unit Patients?
Authors: Akgün Kathleen M. , Murphy Terrence E. , Araujo Katy L. B. , Van Ness Peter H. , Pisani Margaret
Published: October 2010

Introduction. Women receive less aggressive critical care than men based on prior studies. No documented studies evaluate whether men and women are treated equally in the medical intensive care unit (MICU). The Therapeutic Intervention Scoring System-28 (TISS-28) has been used to examine gender differences in mixed ICU studies. However, it has not been used to evaluate equivalence of care in older MICU patients. We hypothesize that given nonsignificant, baseline health differences between genders at MICU admission, the level of care provided would be equivalent.Methods. Prospective cohort of 309 patients≥60 years old in the MICU of an urban university teaching hospital. Explanatory variables were demographic data and baseline measures. Primary outcomes were TISS-28 scores and MICU interventions. We compare TISS-28 scores by gender using a statistical test of equivalence.Results. Women were older and had more chronic respiratory failure at MICU admission. Using equivalence limits of±15% on gender-based scores of TISS-28, MICU interventions were equivalent. Supplementary analysis showed no statistically significant association between gender and mortality.Conclusions. In contrast with other reports from the cardiac critical care literature, as measured by the TISS-28, gender-based care delivered to older MICU patients in this cohort was equivalent.
Definition of Ubiquitination Modulator COP1 as a Novel Therapeutic Target in Human Hepatocellular Carcinoma
Authors: Lee Yun-Han , Andersen Jesper B. , Song Ho-Taek , Judge Adam D. , Seo Daekwan , Ishikawa Tsuyoshi , Marquardt Jens U. , Kitade Mitsuteru , Durkin Marian E. , Raggi Chiara , Woo Hyun Goo , Conner Elizabeth A. , Avital Itzhak , MacLachlan Ian , Factor Valentina M. , Thorgeirsson Snorri S.
Published: October 2010

The development of targeted therapeutics for hepatocellular carcinoma (HCC) remains a major challenge. The ubiquitination modulator COP1 regulates p53 activity by ubiquitination and it is frequently overexpressed in human HCC. In this study, we tested the hypothesis that COP1 blockade by short interfering RNA (siRNA)–mediated inhibition could affect the course of HCC progression. The COP1 isoform COP1-1 was selected as the most effective target for siRNAs in terms of growth inhibition and apoptotic induction in several HCC cell lines. Growth inhibition occurred in HCC cells that retained wild-type p53 or expressed mutant p53 (Y220C or R249S), whereas p53-null Hep3B cells were resistant. Microarray expression analysis revealed that the antiproliferative effects of COP1 blockade were driven by a common subset of molecular alterations including a p53-associated functional network. In an orthotopic mouse xenograft model of HCC, systemic delivery of a modified COP1 siRNA by stable nucleic acid–lipid particles suppressed neoplastic growth in liver without unwanted immune responses. Our findings offer a first proof of principle that COP1 can be a promising target for systemic therapy of HCC. Cancer Res; 70(21); 8264–9. ©2010 AACR.
Overexpression of Transcription Factor Sp2 Inhibits Epidermal Differentiation and Increases Susceptibility to Wound- and Carcinogen-Induced Tumorigenesis
Authors: Kim Tae-Hyung , Chiera Shannon L. , Linder Keith E. , Trempus Carol S. , Smart Robert C. , Horowitz Jonathan M.
Published: October 2010

Sp proteins are evolutionarily conserved transcription factors required for the expression of a wide variety of genes that are critical for development and cell cycle progression. Deregulated expression of certain Sp proteins is associated with the formation of a variety of human tumors; however, direct evidence that any given Sp protein is oncogenic has been lacking. Here, we report that Sp2 protein abundance in mice increases in concert with the progression of carcinogen-induced murine squamous cell carcinomas. Transgenic mice specifically overexpressing murine Sp2 in epidermal basal keratinocytes were highly susceptible to wound- and carcinogen-induced papillomagenesis. Transgenic animals that were homozygous rather than hemizygous for the Sp2 transgene exhibited a striking arrest in the epidermal differentiation program, perishing within 2 weeks of birth. Our results directly support the likelihood that Sp2 overexpression occurring in various human cancers has significant functional effect. Cancer Res; 70(21); 8507–16. ©2010 AACR.
Ablation of TAK1 Upregulates Reactive Oxygen Species and Selectively Kills Tumor Cells
Authors: Omori Emily , Matsumoto Kunihiro , Zhu Songyun , Smart Robert C. , Ninomiya-Tsuji Jun
Published: October 2010

TAK1 kinase activates multiple transcription factors and regulates the level of reactive oxygen species (ROS). We have previously reported that ablation of TAK1 in keratinocytes causes hypersensitivity to ROS-induced cell apoptosis. It is known that some tumor cells produce ROS at higher levels compared with normal cells. We used inducible epidermal-specific TAK1 knockout mice and examined whether ablation of TAK1 in preexisting skin tumors could cause an increase in ROS and result in tumor cell death. Deletion of tak1 gene in skin tumors caused the accumulation of ROS and increased apoptosis, and skin tumors totally regressed within 5 to 10 days. Normal skin did not exhibit any significant abnormality on tak1 gene deletion. Thus, TAK1 kinase could be a new and effective molecular target for ROS-based tumor killing. Cancer Res; 70(21); 8417–25. ©2010 AACR.
Tissue-Specific Pathways for Estrogen Regulation of Ovarian Cancer Growth and Metastasis
Authors: Spillman Monique A. , Manning Nicole G. , Dye Wendy W. , Sartorius Carol A. , Post Miriam D. , Harrell Joshua Chuck , Jacobsen Britta M. , Horwitz Kathryn B.
Published: October 2010

Menopausal estrogen (E2) replacement therapy increases the risk of estrogen receptor (ER)–positive epithelial ovarian cancers (EOC). Whether E2 is tumorigenic or promotes expansion of undiagnosed preexisting disease is unknown. To determine E2 effects on tumor promotion, we developed an intraperitoneal mouse xenograft model using ZsGreen fluorescent ER− 2008 and ER+ PEO4 human EOC cells. Tumor growth was quantified by in vivo fluorescent imaging. In ER+ tumors, E2 significantly increased size, induced progesterone receptors, and promoted lymph node metastasis, confirming that ERs are functional and foster aggressiveness. Laser-captured human EOC cells from ER− and ER+ xenografted tumors were profiled for expression of E2-regulated genes. Three classes of E2-regulated EOC genes were defined, but <10% were shared with E2-regulated breast cancer genes. Because breast cancer selective ER modulators (SERM) are therapeutically ineffective in EOC, we suggest that our EOC-specific E2-regulated genes can assist pharmacologic discovery of ovarian-targeted SERM. Cancer Res; 70(21); 8927–36. ©2010 AACR.
Early Detection of Recurrent Breast Cancer Using Metabolite Profiling
Authors: Asiago Vincent M. , Alvarado Leiddy Z. , Shanaiah Narasimhamurthy , Gowda G.A. Nagana , Owusu-Sarfo Kwadwo , Ballas Robert A. , Raftery Daniel
Published: October 2010

We report on the development of a monitoring test for recurrent breast cancer, using metabolite-profiling methods. Using a combination of nuclear magnetic resonance (NMR) and two-dimensional gas chromatography–mass spectrometry (GC×GC-MS) methods, we analyzed the metabolite profiles of 257 retrospective serial serum samples from 56 previously diagnosed and surgically treated breast cancer patients. One hundred sixteen of the serial samples were from 20 patients with recurrent breast cancer, and 141 samples were from 36 patients with no clinical evidence of the disease during ∼6 years of sample collection. NMR and GC×GC-MS data were analyzed by multivariate statistical methods to compare identified metabolite signals between the recurrence samples and those with no evidence of disease. Eleven metabolite markers (seven from NMR and four from GC×GC-MS) were shortlisted from an analysis of all patient samples by using logistic regression and 5-fold cross-validation. A partial least squares discriminant analysis model built using these markers with leave-one-out cross-validation provided a sensitivity of 86% and a specificity of 84% (area under the receiver operating characteristic curve = 0.88). Strikingly, 55% of the patients could be correctly predicted to have recurrence 13 months (on average) before the recurrence was clinically diagnosed, representing a large improvement over the current breast cancer–monitoring assay CA 27.29. To the best of our knowledge, this is the first study to develop and prevalidate a prediction model for early detection of recurrent breast cancer based on metabolic profiles. In particular, the combination of two advanced analytical methods, NMR and MS, provides a powerful approach for the early detection of recurrent breast cancer. Cancer Res; 70(21); 8309–18. ©2010 AACR.
Cediranib/AZD2171 Inhibits Bone and Brain Metastasis in a Preclinical Model of Advanced Prostate Cancer
Authors: Yin Juan Juan , Zhang Luhua , Munasinghe Jeeva , Linnoila R. Ilona , Kelly Kathleen
Published: October 2010

Late stage or aggressive cancers exhibit metastatic growth at multiple sites, and the characterization of treatment response in various organs to drugs with potentially wide-ranging efficacy is needed. Tumor cells that induce angiogenesis are a common characteristic of metastatic disease, and clinically, antiangiogenic therapies have shown value in the setting of advanced cancer. However, recent preclinical studies have suggested that exposure to antiangiogenic drugs can increase tumor invasiveness and metastasis, making it important to determine which contexts antiangiogenic therapy is most appropriate. We describe here the effects of cediranib, a receptor tyrosine kinase inhibitor, in a model of advanced prostate cancer metastatic to skeleton and brain. Treatment with cediranib decreased metastatic tumor burden in the brain and bone, decreased cerebral vasogenic edema, and improved survival, despite increasing the invasive histology of brain metastases. Short-duration cediranib treatment given at the time of tumor cell dissemination was sufficient to inhibit the establishment and subsequent growth of bone metastases, although brain metastases were subject to rebound growth after the discontinuation of cediranib. Distinct growth patterns at different organ sites in the same animal showed that certain tumor microenvironments such as bone may be most amenable to interventions by anti–vascular endothelial growth factor (VEGF) therapies. In addition, anti-VEGF treatment may be of utility in decreasing the rapid growth of solid brain metastases and vasogenic edema in patients with advanced cancer, leading to reduced morbidity and associated clinical benefit. Cancer Res; 70(21); 8662–73. ©2010 AACR.
K-ras Mutation Targeted to Gastric Tissue Progenitor Cells Results in Chronic Inflammation, an Altered Microenvironment, and Progression to Intraepithelial Neoplasia
Authors: Okumura Tomoyuki , Ericksen Russell E. , Takaishi Shigeo , Wang Sophie S.W. , Dubeykovskiy Zinaida , Shibata Wataru , Betz Kelly S. , Muthupalani Sureshkuma , Rogers Arlin B. , Fox James G. , Rustgi Anil K. , Wang Timothy C.
Published: October 2010

Chronic infectious diseases, such as Helicobacter pylori infection, can promote cancer in a large part through induction of chronic inflammation. Oncogenic K-ras mutation in epithelial cells activates inflammatory pathways, which could compensate for a lack of infectious stimulus. Gastric histopathology and putative progenitor markers [doublecortin and calcium/calmodulin-dependent protein kinase-like 1 (Dcamkl1) and keratin 19 (K19)] in K19-K-ras-V12 (K19-kras) transgenic mice were assessed at 3, 6, 12, and 18 months of age, in comparison with Helicobacter felis–infected wild-type littermates. Inflammation was evaluated by reverse transcription–PCR of proinflammatory cytokines, and K19-kras mice were transplanted with green fluorescent protein (GFP)–labeled bone marrow. Both H. felis infection and K-ras mutation induced upregulation of proinflammatory cytokines, expansion of Dcamkl1+ cells, and progression to oxyntic atrophy, metaplasia, hyperplasia, and high-grade dysplasia. K19-kras transgenic mice uniquely displayed mucous metaplasia as early as 3 months and progressed to high-grade dysplasia and invasive intramucosal carcinoma by 20 months. In bone marrow–transplanted K19-kras mice that progressed to dysplasia, a large proportion of stromal cells were GFP+ and bone marrow–derived, but only rare GFP+ epithelial cells were observed. GFP+ bone marrow–derived cells included leukocytes and CD45− stromal cells that expressed vimentin or α smooth muscle actin and were often found surrounding clusters of Dcamkl1+ cells at the base of gastric glands. In conclusion, the expression of mutant K-ras in K19+ gastric epithelial cells can induce chronic inflammation and promote the development of dysplasia. Cancer Res; 70(21); 8435–45. ©2010 AACR.
Detection of Elevated Plasma Levels of Epidermal Growth Factor Receptor Before Breast Cancer Diagnosis among Hormone Therapy Users
Authors: Pitteri Sharon J. , Amon Lynn M. , Busald Buson Tina , Zhang Yuzheng , Johnson Melissa M. , Chin Alice , Kennedy Jacob , Wong Chee-Hong , Zhang Qing , Wang Hong , Lampe Paul D. , Prentice Ross L. , McIntosh Martin W. , Hanash Samir M. , Li Christopher I.
Published: October 2010

Applying advanced proteomic technologies to prospectively collected specimens from large studies is one means of identifying preclinical changes in plasma proteins that are potentially relevant to the early detection of diseases such as breast cancer. We conducted 14 independent quantitative proteomics experiments comparing pooled plasma samples collected from 420 estrogen receptor–positive (ER+) breast cancer patients ≤17 months before their diagnosis and matched controls. Based on the more than 3.4 million tandem mass spectra collected in the discovery set, 503 proteins were quantified, of which 57 differentiated cases from controls with a P value of <0.1. Seven of these proteins, for which quantitative ELISA assays were available, were assessed in an independent validation set. Of these candidates, epidermal growth factor receptor (EGFR) was validated as a predictor of breast cancer risk in an independent set of preclinical plasma samples for women overall [odds ratio (OR), 1.44; P = 0.0008] and particularly for current users of estrogen plus progestin (E + P) menopausal hormone therapy (OR, 2.49; P = 0.0001). Among current E + P users, the EGFR sensitivity for breast cancer risk was 31% with 90% specificity. Whereas the sensitivity and specificity of EGFR are insufficient for a clinically useful early detection biomarker, this study suggests that proteins that are elevated preclinically in women who go on to develop breast cancer can be discovered and validated using current proteomic technologies. Further studies are warranted to examine the role of EGFR and to discover and validate other proteins that could potentially be used for early detection of breast cancer. Cancer Res; 70(21); 8598–606. ©2010 AACR.
F3-Targeted Cisplatin-Hydrogel Nanoparticles as an Effective Therapeutic That Targets Both Murine and Human Ovarian Tumor Endothelial Cells In vivo
Authors: Winer Ira , Wang Shouyan , Lee Youg-Eun Koo , Fan Wenzhe , Gong Yusong , Burgos-Ojeda Daniela , Spahlinger Greg , Kopelman R. , Buckanovich Ronald J.
Published: October 2010

Recent studies indicate that ovarian cancer may be highly responsive to antivascular therapeutics. We have developed an antivascular tumor therapeutic using the F3 peptide to target cisplatin-loaded nanoparticles (F3-Cis-Np) to tumor vessels. We show that although F3-Cis-Np bind with high specificity to both human ovarian tumor cells and tumor endothelial cells in vitro, they only show cytotoxic activity against the tumor endothelial cells. In vivo these nanoparticles bind primarily to tumor endothelial cells. Therapeutic studies in both flank and orthotopic i.p. murine ovarian tumor models, as well as human tumor xenograft models, show rapid tumor regression with treatment. Treatment was associated with significant vascular necrosis consistent with an antivascular effect. Furthermore, treatment was active in both platinum-sensitive and platinum-resistant cell lines. Importantly, we show that F3-Cis-Np bind to human tumor endothelial cells in vitro and to human tumor vessels in vivo. Therapy targeting human vasculature in vivo with F3-Cis-Np led to near complete loss of all human tumor vessels in a murine model of human tumor vasculature. Our studies indicate that F3-targeted vascular therapeutics may be an effective treatment modality in human ovarian cancer. Cancer Res; 70(21); 8674–83. ©2010 AACR.
Antibodies to Merkel Cell Polyomavirus T Antigen Oncoproteins Reflect Tumor Burden in Merkel Cell Carcinoma Patients
Authors: Paulson Kelly G. , Carter Joseph J. , Johnson Lisa G. , Cahill Kevin W. , Iyer Jayasri G. , Schrama David , Becker Juergen C. , Madeleine Margaret M. , Nghiem Paul , Galloway Denise A.
Published: October 2010

Merkel cell polyomavirus (MCPyV) is a common infectious agent that is likely involved in the etiology of most Merkel cell carcinomas (MCC). Serum antibodies recognizing the MCPyV capsid protein VP1 are detectable at high titer in nearly all MCC patients and remain stable over time. Although antibodies to the viral capsid indicate prior MCPyV infection, they provide limited clinical insight into MCC because they are also detected in more than half of the general population. We investigated whether antibodies recognizing MCPyV large and small tumor-associated antigens (T-Ag) would be more specifically associated with MCC. Among 530 population control subjects, these antibodies were present in only 0.9% and were of low titer. In contrast, among 205 MCC cases, 40.5% had serum IgG antibodies that recognize a portion of T-Ag shared between small and large T-Ags. Among cases, titers of T-Ag antibodies fell rapidly (∼8-fold per year) in patients whose cancer did not recur, whereas they rose rapidly in those with progressive disease. Importantly, in several patients who developed metastases, the rise in T-Ag titer preceded clinical detection of disease spread. These results suggest that antibodies recognizing T-Ag are relatively specifically associated with MCC, do not effectively protect against disease progression, and may serve as a clinically useful indicator of disease status. Cancer Res; 70(21); 8388–97. ©2010 AACR.
TheIFNG(IFN-γ) Genotype Predicts Cytogenetic and Molecular Response to Imatinib Therapy in Chronic Myeloid Leukemia
Authors: Kim Dong Hwan (Dennis) , Kong Jee Hyun , Byeun Ji Young , Jung Chul Won , Xu Wei , Liu Xiangdong , Kamel-Reid Suzanne , Kim Yeo-Keyoung , Kim Hyeoung-Joon , Lipton Jeffrey H.
Published: October 2010

Purpose: The present study analyzed treatment outcomes of imatinib therapy by interindividual genetic variants in candidate biological pathways of chronic myeloid leukemia (CML) such as apoptosis, angiogenesis, IFN-γ signaling pathways, or drug transport/metabolism of imatinib.

Experimental Design: Peripheral blood DNAs were genotyped for 79 single nucleotide polymorphism markers involved in the pathways of apoptosis, angiogenesis, myeloid cell growth, xenobiotic metabolism, WT1 signaling, IFN signaling, and others in CML patients who were included in discovery (n = 229, Canada) and validation cohorts (n = 187, Korea).

Results: We found several genotypes associated with complete cytogenetic response: IFNG (rs1861494, rs2069705), FASL (rs763110), FAS (rs2234767, rs2234978), VEGFR2 (rs1531289), and WT1 (rs2234590); with major molecular response: IFNG (rs1861494, rs2069705), BIRC5 (rs9904341), FAS (rs2234978), and ABCG2 (rs2231142); with loss of response: IFNG (rs2069705), IFNGR2 (rs9808753), BIRC5 (rs9904341), and ORM (rs3182041); and with treatment failure: IFNG (rs2069705), JAK3 (rs3212713), and ORM (rs3182041). External validation for the above significant genotypes confirmed that the IFNG genotype (rs2069705) was predictive of complete cytogenetic response (hazard ratio, 2.17; P < 0.001) and major molecular response (hazard ratio, 1.96; P = 0.0001) in validation cohorts of Korean ethnicity.

Conclusions: The IFNG genotype was predictive for response to imatinib therapy, suggesting potential involvement of the IFN-γ signaling pathway in the mechanism of action of imatinib in CML. Clin Cancer Res; 16(21); 5339–50. ©2010 AACR.
Safety and Clinical Effect of Subcutaneous Human Interleukin-21 in Patients with Metastatic Melanoma or Renal Cell Carcinoma: A Phase I Trial
Authors: Schmidt Henrik , Brown Janet , Mouritzen Ulrik , Selby Peter , Fode Kirsten , Svane Inge Marie , Cook Graham P. , Mollerup David Hal , Geertsen Poul F.
Published: October 2010

Purpose: This phase I study in patients with metastatic melanoma (MM) and renal cell carcinoma (RCC) evaluated the safety and maximum tolerated dose (MTD), pharmacokinetics, pharmacodynamics, and preliminary antitumor activity of s.c. treatment of human recombinant interleukin 21 (IL-21).

Experimental Design: Phase I dose-escalation trial with treatment of three to six patients at each dose level, escalating from 3 to 300 μg/kg. Treatment was administered s.c. on an outpatient basis 3 days per week for 8 or 16 weeks.

Results: Twenty-six patients entered the study. Recombinant IL-21 was generally well tolerated, and dose-limiting toxicities (DLT) were first seen at dose levels of 200 and 300 μg/kg. The following four DLTs were observed in three patients: increased transaminases, increased hyperbilirubinemia, hypersensitivity reaction, and lethargy. The MTD was declared to be 200 μg/kg, although five of seven patients at the 300 μg/kg dose level experienced no DLTs. A treatment-related effect on soluble CD25 was observed at all dose levels and increased with dose level. Furthermore, higher doses induced interferon-γ, perforin, and granzyme B mRNA expression in peripheral blood, and granzyme B protein expression in both CD8+ T cells and natural killer cells, consistent with the activation of cytotoxic lymphocytes. Three patients, one patient with MM and two with RCC, obtained a partial response.

Conclusion: Outpatient treatment with s.c. administered IL-21 was tolerated and had dose-dependent pharmacodynamics. rIL-21 showed antitumor activity in patients with MM and RCC. Clin Cancer Res; 16(21); 5312–9. ©2010 AACR.