bluuebunny/crossref_metadata_2025 huggingface dataset:
MicroRNA-663 upregulated by oscillatory shear stress plays a role in inflammatory response of endothelial cells
Authors: Ni Chih-Wen , Qiu Haiwei , Jo Hanjoong
Published: March 2011

The mechanisms by which oscillatory shear stress (OS) induces, while high laminar shear stress (LS) prevents, atherosclerosis are still unclear. Here, we examined the hypothesis that OS induces inflammatory response, a critical atherogenic event, in endothelial cells by a microRNA (miRNA)-dependent mechanism. By miRNA microarray analysis using total RNA from human umbilical vein endothelial cells (HUVECs) that were exposed to OS or LS for 24 h, we identified 21 miRNAs that were differentially expressed. Of the 21 miRNAs, 13 were further examined by quantitative PCR, which validated the result for 10 miRNAs. Treatment of HUVECs with the miR-663 antagonist (miR-663-locked nucleic acids) blocked OS-induced monocyte adhesion, but not apoptosis. In contrast, overexpression of miR-663 increased monocyte adhesion in LS-exposed cells. Subsequent mRNA expression microarray study using HUVECs treated with miR-663-locked nucleic acids and OS revealed 32 up- and 3 downregulated genes, 6 of which are known to be involved in inflammatory response. In summary, we identified 10 OS-sensitive miRNAs, including miR-663, which plays a key role in OS-induced inflammatory responses by mediating the expression of inflammatory gene network in HUVECs. These OS-sensitive miRNAs may mediate atherosclerosis induced by disturbed flow.
Critical role of CFTR-dependent lipid rafts in cigarette smoke-induced lung epithelial injury
Authors: Bodas Manish , Min Taehong , Vij Neeraj
Published: March 2011

Apoptosis of lung epithelial and endothelial cells by exposure to cigarette smoke (CS) severely damages the lung tissue, leading to the pathogenesis of emphysema, but the underlying mechanisms are poorly understood. We have recently established a direct correlation between decreased lipid raft CFTR expression and emphysema progression through increased ceramide accumulation. In the present work, we investigated the role of membrane CFTR in regulating apoptosis and autophagy responses to CS exposure. We report a constitutive and CS-induced increase in the number of TUNEL-positive apoptotic cells in Cftr−/− murine lungs compared with Cftr+/+ murine lungs that also correlated with a concurrent increase in the expression of ceramide, NF-κB, CD95/Fas, lipid raft proteins, and zonula occludens (ZO)-1/2 ( P < 0.001). We also verified that stable wild-type CFTR expression in CFBE41o− cells controls constitutively elevated caspase-3/7 activity (−1.6-fold, P < 0.001). Our data suggest that membrane CFTR regulates ceramide-enriched lipid raft signaling platforms required for the induction of Fas-mediated apoptotic signaling. In addition, lack of membrane CFTR also modulates autophagy, as demonstrated by the significant increase in constitutive ( P < 0.001) and CSE-induced ( P < 0.005) perinuclear accumulation of green fluorescent protein-microtubule-associated protein 1 light chain-3 (LC3) in the absence of membrane CFTR (CFBE41o− cells). The significant constitutive and CS-induced increase ( P < 0.05) in p62 and LC3β expression in CFTR-deficient cells and mice corroborates these findings and suggest a defective autophagy response in the absence of membrane CFTR. Our data demonstrate the critical role of membrane-localized CFTR in regulating apoptotic and autophagic responses in CS-induced lung injury that may be involved in the pathogenesis of severe emphysema.
Cerebral autoregulation dynamics in endurance-trained individuals
Authors: Lind-Holst Mikkel , Cotter James D. , Helge Jørn W. , Boushel Robert , Augustesen Helene , Van Lieshout Johannes J. , Pott Frank C.
Published: March 2011

Aerobic fitness may be associated with reduced orthostatic tolerance. To investigate whether trained individuals have less effective regulation of cerebral vascular resistance, we studied the middle cerebral artery (MCA) mean blood velocity ( Vmean) response to a sudden drop in mean arterial pressure (MAP) after 2.5 min of leg ischemia in endurance athletes and untrained subjects (maximal O2uptake: 69 ± 7 vs. 42 ± 5 ml O2·min−1·kg−1; n = 9 for both, means ± SE). After cuff release when seated, endurance athletes had larger drops in MAP (94 ± 6 to 62 ± 5 mmHg, −39%, vs. 99 ± 5 to 73 ± 4 mmHg, −26%) and MCA Vmean(53 ± 3 to 37 ± 2 cm/s, −30%, vs. 58 ± 3 to 43 ± 2 cm/s, −25%). The athletes also had a slower recovery to baseline of both MAP (25 ± 2 vs. 16 ± 1 s, P < 0.01) and MCA Vmean(15 ± 1 vs. 11 ± 1 s, P < 0.05). The onset of autoregulation, determined by the time point of increase in the cerebrovascular conductance index (CVCi = MCA Vmean/MAP) appeared later in the athletes (3.9 ± 0.4 vs. 2.7 ± 0.4s, P = 0.01). Spectral analysis revealed a normal MAP-to-MCA Vmeanphase in both groups but ∼40% higher normalized MAP to MCA Vmeanlow-frequency transfer function gain in the trained subjects. No significant differences were detected in the rates of recovery of MAP and MCA Vmeanand the rate of CVCi regulation (18 ± 4 vs. 24 ± 7%/s, P = 0.2). In highly trained endurance athletes, a drop in blood pressure after the release of resting leg ischemia was more pronounced than in untrained subjects and was associated with parallel changes in indexes of cerebral blood flow. Once initiated, the autoregulatory response was similar between the groups. A delayed onset of autoregulation with a larger normalized transfer gain conforms with a less effective dampening of MAP oscillations, indicating that athletes may be more prone to instances of symptomatic cerebral hypoperfusion when MAP declines.
Crystal structure of layered perovskite compound, Li2LaTa2O6N
Authors: Kaga Motoaki , Kurachi Hirokazu , Asaka Toru , Yue Bing , Ye Jinhua , Fukuda Koichiro
Published: March 2011

The crystal structure of Li2LaTa2O6N was determined from laboratory X-ray powder diffraction data (CuKα1) using the Rietveld method. The title compound is tetragonal with space groupI4/mmm,Z=2, and unit-cell dimensionsa=0.395 049(4) nm,c=1.850 97(3) nm, andV=0.288 869(6) nm3. The initial structural model was successfully derived by the direct methods and further refined by the Rietveld method, with the anisotropic atomic displacement parameters being assigned for all atoms. The final reliability indices wereRwp=5.73%,S=1.46,Rp=4.33%,RB=1.13%, andRF=0.53%. Li2LaTa2O6N has a layered perovskite structure similar to that of Li2LaTa2O7.
Typical values of Rietveld instrument profile coefficients
Authors: Kaduk James A. , Reid Joel
Published: March 2011

GSASinstrument parameters are tabulated for a variety of laboratory and synchrotron diffractometers to give users an idea of the typical ranges of profile parameters when they generate their own instrument parameter files. For modern high-resolution laboratory diffractometers, the parameters fall in the ranges 0<U<3,V=0, 0<W<4, 1<X<3, 0<Y<3, 1<asym<3, and 0<S/L<0.03. For synchrotron diffractometers, the parameters fall in the ranges 0<U<1.2, −1<V<0, 0<W<1, 0<X<1, 0<Y<1, 0<asym<0.5, 0<S/L<0.001, and 0<H/L<0.007.FULLPROFequivalents are also reported. The factors which are convoluted together to generate the instrument profile are described.
Does Ordinary Portland Cement contain amorphous phase? A quantitative study using an external standard method
Authors: Jansen D. , Stabler Ch. , Goetz-Neunhoeffer F. , Dittrich S. , Neubauer J.
Published: March 2011

A suitable external standard method which was first described by O’Connor and Raven (1988) (“Application of the Rietveld refinement procedure in assaying powdered mixtures,” Powder Diffr. 3, 2–6) was used to determine the quantitative phase composition of a commonly used Ordinary Portland Cement (OPC). The method was also applied in order to determine amorphous contents in OPC. Also investigated were the impact of atomic displacement parameters and the microstrain on the calculated amorphous content. The investigations yielded evidence that said parameters do indeed exert an influence on the calculated amorphous content. On the basis of the data produced we can conclude that the method used is entirely to be recommended for the examination of OPC. No significant amorphous content could be proven in the OPC used.
Microtwinning hypothesis for a more ordered vaterite model
Authors: Le Bail A. , Ouhenia S. , Chateigner D.
Published: March 2011

An orthorhombic fully ordered structural model is proposed for vaterite [space group Ama2, a=8.4721(5) Å, b=7.1575(7) Å, c=4.1265(4) Å, Z=4, and V=250.23(4) Å3]. It is based on a microtwinning hypothesis, with three domains rotated by 120° along the orthorhombic a axis, regenerating a pseudohexagonal habit. The solution came from direct space ab initio calculations applied to the powder diffraction data. However, five weak superstructure reflections seen in single-crystal and powder diffraction experiments, leading to a six times larger unit cell, are still unexplained.
Chemical preparation and XRD data for a new triphosphate CuNa3P3O10 and two cyclotriphosphates SrRbP3O9·3H2O and SrRbP3O9
Authors: Fahim I. , Kheïreddine A. , Tridane M. , Belaaouad S.
Published: March 2011

Methods of chemical preparation and XRD data are reported for a new triphosphate CuNa3P3O10 and two cyclotriphosphates SrRbP3O9·3H2O and SrRbP3O9. SrRbP3O9·3H2O was prepared by the method of ion-exchange resin, while CuNa3P3O10 and SrRbP3O9 were obtained by total dehydration of CuNa3P3O10·12H2O and SrRbP3O9·3H2O, respectively. CuNa3P3O10 crystallizes in the hexagonal system, with space group P-31c, Z=2, and the following unit-cell dimensions: a=b=7.022(1) Å, c=9.217(1) Å, M(20)=81, F(20)=117(0.003 419, 50), and V=393.24(2) Å3. SrRbP3O9·3H2O is orthorhombic, with Z=4, space group Pnma, and the following unit-cell dimensions: a=9.120(1) Å, b=8.141(1) Å, c=15.234(1) Å, M(20)=5.1, F(20)=5.8(0.0173,199), and V=1131.1(3) Å3. SrRbP3O9 is monoclinic, with space group P21/m or P21, Z=4, and the following unit-cell dimensions: a=14.958(3) Å, b=8.503(2) Å, c=7.898(2) Å, β=122.19(2)°, M(20)=9.9, F(20)=16.5(0.0189, 64), and V=850.2(8) Å3.
Troponin I and C-Reactive Protein Are Commonly Detected in Patients with Breast Cancer Treated with Dose-Dense Chemotherapy Incorporating Trastuzumab and Lapatinib
Authors: Morris Patrick G. , Chen Carol , Steingart Richard , Fleisher Martin , Lin Nancy , Moy Beverly , Come Steven , Sugarman Steven , Abbruzzi Alyson , Lehman Robert , Patil Sujata , Dickler Maura , McArthur Heather L. , Winer Eric , Norton Larry , Hudis Clifford A. , Dang Chau T.
Published: March 2011

Purpose: There are no validated methods of early detection of cardiotoxicity from trastuzumab (T) following anthracycline-based chemotherapy. Currently changes in left ventricular ejection fraction (LVEF) are assessed but this approach has limited sensitivity and specificity. Within a prospective feasibility study of dose-dense (dd) doxorubicin and cyclophosphamide (AC) → weekly paclitaxel (P) with T and lapatinib (L), we included a preplanned analysis of correlative cardiac Troponin I (cTnI) and C-reactive protein (CRP) as early biomarkers of cardiotoxicity.

Experimental Design: As previously described, patients received ddACx 4→PTL→TL. LVEF was assessed at months 0, 2, 6, 9, and 18 and cTnI and CRP measured every 2 weeks during chemotherapy then at months 6, 9, and 18. These biomarkers were correlated with changes in LVEF.

Results: Ninety-five patients enrolled. Overall, 3 (3%) patients withdrew during AC and 41 (43%) withdrew during PTL→TL, mostly due to diarrhea. Median LVEF was 68% (baseline), 69% (month 2), 65% (month 6), 65% (month 9), and 65% (month 18). The majority (67%) had a detectable cTnI during the study. The proportion of detectable cTnIs increased over time; 4% at baseline, 11% at month 2, and 50% at month 3. The timing of these detectable cTnIs preceded maximum-recorded decline in LVEF. However, overall, maximum cTnI levels did not correlate with LVEF declines. A detectable CRP was seen in 74/95 (78%) but did not correlate with LVEF declines.

Conclusion: In patients receiving ddAC→PTL, cTnIs are commonly detected. These elevations may precede changes in LVEF but, as assessed in this trial, do not predict CHF. Clin Cancer Res; 17(10); 3490–9. ©2011 AACR.
Dual Targeting of Phosphoinositide 3-Kinase and Mammalian Target of Rapamycin Using NVP-BEZ235 as a Novel Therapeutic Approach in Human Ovarian Carcinoma
Authors: Santiskulvong Chintda , Konecny Gottfried E. , Fekete Mirela , Chen Kuang-Yui Michael , Karam Amer , Mulholland David , Eng Carol , Wu Hong , Song Min , Dorigo Oliver
Published: March 2011

Purpose: This study evaluates the effect of dual PI3K and mTOR inhibition using NVP-BEZ235 in preclinical models of ovarian cancer as a potential novel therapeutic strategy.

Experimental Design: Inhibition of PI3K/Akt/mTOR signaling by NVP-BEZ235 was demonstrated by immunoblotting. The effect on cell proliferation was assessed in 18 ovarian cancer cell lines, including four pairs of syngeneic cisplatin-sensitive and cisplatin-resistant cell lines. The in vivo effects of NVP-BEZ235 on established tumor growth were evaluated using an immunocompetent, transgenic murine ovarian cancer model (LSL-K-rasG12D/+PtenloxP/loxP).

Results: NVP-BEZ235 decreased cell proliferation in all ovarian cancer cell lines assayed and sensitized cisplatin-resistant cells to the cytotoxic effects of cisplatin. Cell lines with PI3K-activating mutations or Pten deletions were significantly more sensitive to the effect of NVP-BEZ235 than cell lines without these mutations (P < 0.05). A statistically significant correlation was found between relative levels of p4E-BP1 and the IC50 for NVP-BEZ235. In LSL-K-rasG12D/+PtenloxP/loxP mice with established intraperitoneal tumor disease, oral administration of NVP-BEZ235 decreased pAkt, p4E-BP1 and Ki67 in tumor tissue, and resulted in significantly longer survival compared to control animals (P < 0.05). NVP-BEZ235 also induced cell cycle arrest, caspase 3 activity, and reduced cell migration.

Conclusions: Targeting PI3K and mTOR simultaneously using NVP-BEZ235 effectively inhibits ovarian cancer cell growth even in the presence of platinum resistance and prolongs survival of mice with intra-abdominal ovarian tumor disease. We propose that dual PI3K and mTOR inhibition using NVP-BEZ235 may be an effective novel therapeutic approach in patients with ovarian cancer. Clin Cancer Res; 17(8); 2373–84. ©2011 AACR.
Epstein-Barr Virus–Associated B-cell Lymphomas: Pathogenesis and Clinical Outcomes
Authors: Saha Abhik , Robertson Erle S.
Published: March 2011

Epstein-Barr virus (EBV) is a ubiquitous human γ-herpesvirus that establishes a life-long asymptomatic infection in immunocompetent hosts. It is also found to be frequently associated with a broad spectrum of B-cell lymphomas predominantly seen in immunodeficient patients. Despite many resemblances, these EBV-linked lymphoproliferative disorders display heterogeneity at the clinical and the molecular level. Moreover, EBV-associated lymphoproliferative diseases differ in their differential expression patterns of the EBV-encoded latent antigens, which are directly related to their interactions with the host. EBV-driven primary B-cell immortalization is linked to the cooperative functions of these latent proteins, which are critical for perturbing many important cell-signaling pathways maintaining B-cell proliferation. Additionally, it is used as a surrogate model to explore the underlying mechanisms involved in the development of B-cell neoplasms. Recent discoveries have revealed that a number of sophisticated mechanisms are exploited by EBV during cancer progression. This finding will be instrumental in the design of novel approaches for therapeutic interventions against EBV-associated B-cell lymphomas. This review limits the discussion to the biology and pathogenesis of EBV-associated B-cell lymphomas and the related clinical implications. Clin Cancer Res; 17(10); 3056–63. ©2011 AACR.
Humanized Anti-Trop-2 IgG-SN-38 Conjugate for Effective Treatment of Diverse Epithelial Cancers: Preclinical Studies in Human Cancer Xenograft Models and Monkeys
Authors: Cardillo Thomas M. , Govindan Serengulam V. , Sharkey Robert M. , Trisal Preeti , Goldenberg David M.
Published: March 2011

Purpose: Evaluate the efficacy of an SN-38-anti-Trop-2 antibody–drug conjugate (ADC) against several human solid tumor types, and to assess its tolerability in mice and monkeys, the latter with tissue cross-reactivity to hRS7 similar to humans.

Experimental Design: Two SN-38 derivatives, CL2-SN-38 and CL2A-SN-38, were conjugated to the anti-Trop-2–humanized antibody, hRS7. The immunoconjugates were characterized in vitro for stability, binding, and cytotoxicity. Efficacy was tested in five different human solid tumor-xenograft models that expressed Trop-2 antigen. Toxicity was assessed in mice and in Cynomolgus monkeys.

Results: The hRS7 conjugates of the two SN-38 derivatives were equivalent in drug substitution (∼6), cell binding (Kd ∼ 1.2 nmol/L), cytotoxicity (IC50 ∼ 2.2 nmol/L), and serum stability in vitro (t/½ ∼ 20 hours). Exposure of cells to the ADC demonstrated signaling pathways leading to PARP cleavage, but differences versus free SN-38 in p53 and p21 upregulation were noted. Significant antitumor effects were produced by hRS7-SN-38 at nontoxic doses in mice bearing Calu-3 (P ≤ 0.05), Capan-1 (P < 0.018), BxPC-3 (P < 0.005), and COLO 205 tumors (P < 0.033) when compared to nontargeting control ADCs. Mice tolerated a dose of 2 × 12 mg/kg (SN-38 equivalents) with only short-lived elevations in ALT and AST liver enzyme levels. Cynomolgus monkeys infused with 2 × 0.96 mg/kg exhibited only transient decreases in blood counts, although, importantly, the values did not fall below normal ranges.

Conclusions: The anti-Trop-2 hRS7-CL2A-SN-38 ADC provides significant and specific antitumor effects against a range of human solid tumor types. It is well tolerated in monkeys, with tissue Trop-2 expression similar to humans, at clinically relevant doses, and warrants clinical investigation. Clin Cancer Res; 17(10); 3157–69. ©2011 AACR.
Therapeutic Potential and Molecular Mechanism of a Novel, Potent, Nonpeptide, Smac Mimetic SM-164 in Combination with TRAIL for Cancer Treatment
Authors: Lu Jianfeng , McEachern Donna , Sun Haiying , Bai Longchuan , Peng Yuefeng , Qiu Su , Miller Rebecca , Liao Jinhui , Yi Han , Liu Meilan , Bellail Anita , Hao Chunhai , Sun Shi-Yong , Ting Adrian T. , Wang Shaomeng
Published: March 2011

Smac mimetics are being developed as a new class of anticancer therapies. Because the single-agent activity of Smac mimetics is very limited, rational combinations represent a viable strategy for their clinical development. The combination of Smac mimetics with TNF-related apoptosis inducing ligand (TRAIL) may be particularly attractive because of the low toxicity of TRAIL to normal cells and the synergistic antitumor activity observed for the combination. In this study, we have investigated the combination synergy between TRAIL and a potent Smac mimetic, SM-164, in vitro and in vivo and the underlying molecular mechanism of action for the synergy. Our study shows that SM-164 is highly synergistic with TRAIL in vitro in both TRAIL-sensitive and TRAIL-resistant cancer cell lines of breast, prostate, and colon cancer. Furthermore, the combination of SM-164 with TRAIL induces rapid tumor regression in vivo in a breast cancer xenograft model in which either agent is ineffective. Our data show that X-linked IAP (XIAP) and cellular IAP 1 (cIAP1), but not cIAP2, work in concert to attenuate the activity of TRAIL; SM-164 strongly enhances TRAIL activity by concurrently targeting XIAP and cIAP1. Moreover, although RIP1 plays a minimal role in the activity of TRAIL as a single agent, it is required for the synergistic interaction between TRAIL and SM-164. This study provides a strong rationale to develop the combination of SM-164 and TRAIL as a new therapeutic strategy for the treatment of human cancer. Mol Cancer Ther; 10(5); 902–14. ©2011 AACR.
Phase IIa Clinical Trial of Curcumin for the Prevention of Colorectal Neoplasia
Authors: Carroll Robert E. , Benya Richard V. , Turgeon Danielle Kim , Vareed Shaiju , Neuman Malloree , Rodriguez Luz , Kakarala Madhuri , Carpenter Philip M. , McLaren Christine , Meyskens Frank L. , Brenner Dean E.
Published: March 2011

Curcumin is derived from the spice tumeric and has antiinflammatory and antineoplastic effects in vitro and in animal models, including preventing aberrant crypt foci (ACF) and adenomas in murine models of colorectal carcinogenesis. Inhibiting the production of the procarcinogenic eicosanoids prostaglandin E2 (PGE2) and 5-hydroxyeicosatetraenoic acid (5-HETE) can suppress carcinogenesis in rodents. Curcumin reduces mucosal concentrations of PGE2 (via inhibition of cyclooxygenases 1 and 2) and 5-HETE (via inhibition of 5-lipoxygenase) in rats. Although preclinical data support curcumin activity in many sites, the poor bioavailability reported for this agent supports its use in the colorectum. We assessed the effects of oral curcumin (2 g or 4 g per day for 30 days) on PGE2 within ACF (primary endpoint), 5-HETE, ACF number, and proliferation in a nonrandomized, open-label clinical trial in 44 eligible smokers with eight or more ACF on screening colonoscopy. We assessed pre- and posttreatment concentrations of PGE2 and 5-HETE by liquid chromatography tandem mass spectroscopy in ACF and normal-tissue biopsies; ACF number via rectal endoscopy; proliferation by Ki-67 immunohistochemistry; and curcumin concentrations by high-performance liquid chromatography in serum and rectal mucosal samples. Forty-one subjects completed the study. Neither dose of curcumin reduced PGE2 or 5-HETE within ACF or normal mucosa or reduced Ki-67 in normal mucosa. A significant 40% reduction in ACF number occurred with the 4-g dose (P < 0.005), whereas ACF were not reduced in the 2-g group. The ACF reduction in the 4-g group was associated with a significant, five-fold increase in posttreatment plasma curcumin/conjugate levels (versus pretreatment; P = 0.009). Curcumin was well tolerated at both 2 g and 4 g. Our data suggest that curcumin can decrease ACF number, and this is potentially mediated by curcumin conjugates delivered systemically. Cancer Prev Res; 4(3); 354–64. ©2011 AACR.
A Framework for Evaluating Biomarkers for Early Detection: Validation of Biomarker Panels for Ovarian Cancer
Authors: Zhu Claire S. , Pinsky Paul F. , Cramer Daniel W. , Ransohoff David F. , Hartge Patricia , Pfeiffer Ruth M. , Urban Nicole , Mor Gil , Bast Robert C. , Moore Lee E. , Lokshin Anna E. , McIntosh Martin W. , Skates Steven J. , Vitonis Allison , Zhang Zhen , Ward David C. , Symanowski James T. , Lomakin Aleksey , Fung Eric T. , Sluss Patrick M. , Scholler Nathalie , Lu Karen H. , Marrangoni Adele M. , Patriotis Christos , Srivastava Sudhir , Buys Saundra S. , Berg Christine D.
Published: March 2011

A panel of biomarkers may improve predictive performance over individual markers. Although many biomarker panels have been described for ovarian cancer, few studies used prediagnostic samples to assess the potential of the panels for early detection. We conducted a multisite systematic evaluation of biomarker panels using prediagnostic serum samples from the Prostate, Lung, Colorectal, and Ovarian Cancer (PLCO) screening trial.

Using a nested case–control design, levels of 28 biomarkers were measured laboratory-blinded in 118 serum samples obtained before cancer diagnosis and 951 serum samples from matched controls. Five predictive models, each containing 6 to 8 biomarkers, were evaluated according to a predetermined analysis plan. Three sequential analyses were conducted: blinded validation of previously established models (step 1); simultaneous split-sample discovery and validation of models (step 2); and exploratory discovery of new models (step 3). Sensitivity, specificity, sensitivity at 98% specificity, and AUC were computed for the models and CA125 alone among 67 cases diagnosed within one year of blood draw and 476 matched controls. In step 1, one model showed comparable performance to CA125, with sensitivity, specificity, and AUC at 69.2%, 96.6%, and 0.892, respectively. Remaining models had poorer performance than CA125 alone. In step 2, we observed a similar pattern. In step 3, a model derived from all 28 markers failed to show improvement over CA125.

Thus, biomarker panels discovered in diagnostic samples may not validate in prediagnostic samples; utilizing prediagnostic samples for discovery may be helpful in developing validated early detection panels. Cancer Prev Res; 4(3); 375–83. ©2011 AACR.
Bioavailability of Sulforaphane from Two Broccoli Sprout Beverages: Results of a Short-term, Cross-over Clinical Trial in Qidong, China
Authors: Egner Patricia A. , Chen Jian Guo , Wang Jin Bing , Wu Yan , Sun Yan , Lu Jian Hua , Zhu Jian , Zhang Yong Hui , Chen Yong Sheng , Friesen Marlin D. , Jacobson Lisa P. , Muñoz Alvaro , Ng Derek , Qian Geng Sun , Zhu Yuan Rong , Chen Tao Yang , Botting Nigel P. , Zhang Qingzhi , Fahey Jed W. , Talalay Paul , Groopman John D , Kensler Thomas W.
Published: March 2011

One of several challenges in design of clinical chemoprevention trials is the selection of the dose, formulation, and dose schedule of the intervention agent. Therefore, a cross-over clinical trial was undertaken to compare the bioavailability and tolerability of sulforaphane from two of broccoli sprout–derived beverages: one glucoraphanin-rich (GRR) and the other sulforaphane-rich (SFR). Sulforaphane was generated from glucoraphanin contained in GRR by gut microflora or formed by treatment of GRR with myrosinase from daikon (Raphanus sativus) sprouts to provide SFR. Fifty healthy, eligible participants were requested to refrain from crucifer consumption and randomized into two treatment arms. The study design was as follows: 5-day run-in period, 7-day administration of beverages, 5-day washout period, and 7-day administration of the opposite intervention. Isotope dilution mass spectrometry was used to measure levels of glucoraphanin, sulforaphane, and sulforaphane thiol conjugates in urine samples collected daily throughout the study. Bioavailability, as measured by urinary excretion of sulforaphane and its metabolites (in approximately 12-hour collections after dosing), was substantially greater with the SFR (mean = 70%) than with GRR (mean = 5%) beverages. Interindividual variability in excretion was considerably lower with SFR than with GRR beverage. Elimination rates were considerably slower with GRR, allowing for achievement of steady-state dosing as opposed to bolus dosing with SFR. Optimal dosing formulations in future studies should consider blends of sulforaphane and glucoraphanin as SFR and GRR mixtures to achieve peak concentrations for activation of some targets and prolonged inhibition of others implicated in the protective actions of sulforaphane. Cancer Prev Res; 4(3); 384–95. ©2011 AACR.
Obesity Is Associated with Inflammation and Elevated Aromatase Expression in the Mouse Mammary Gland
Authors: Subbaramaiah Kotha , Howe Louise R. , Bhardwaj Priya , Du Baoheng , Gravaghi Claudia , Yantiss Rhonda K. , Zhou Xi Kathy , Blaho Victoria A. , Hla Timothy , Yang Peiying , Kopelovich Levy , Hudis Clifford A. , Dannenberg Andrew J.
Published: March 2011

Elevated circulating estrogen levels are associated with increased risk of breast cancer in obese postmenopausal women. Following menopause, the biosynthesis of estrogens through CYP19 (aromatase)-mediated metabolism of androgen precursors occurs primarily in adipose tissue, and the resulting estrogens are then secreted into the systemic circulation. The potential links between obesity, inflammation, and aromatase expression are unknown. In both dietary and genetic models of obesity, we observed necrotic adipocytes surrounded by macrophages forming crown-like structures (CLS) in the mammary glands and visceral fat. The presence of CLS was associated with activation of NF-κB and increased levels of proinflammatory mediators (TNF-α, IL-1β, Cox-2), which were paralleled by elevated levels of aromatase expression and activity in the mammary gland and visceral fat of obese mice. Analyses of the stromal-vascular and adipocyte fractions of the mammary gland suggested that macrophage-derived proinflammatory mediators induced aromatase and estrogen-dependent gene expression (PR, pS2) in adipocytes. Saturated fatty acids, which have been linked to obesity-related inflammation, stimulated NF-κB activity in macrophages leading to increased levels of TNF-α, IL-1β, and Cox-2, each of which contributed to the induction of aromatase in preadipocytes. The discovery of the obesity → inflammation → aromatase axis in the mammary gland and visceral fat and its association with CLS may provide insight into mechanisms underlying the increased risk of hormone receptor-positive breast cancer in obese postmenopausal women, the reduced efficacy of aromatase inhibitors in the treatment of breast cancer in these women, and their generally worse outcomes. The presence of CLS may be a biomarker of increased breast cancer risk or poor prognosis. Cancer Prev Res; 4(3); 329–46. ©2011 AACR.
Curcumin Chemoprevention: The Long Road to Clinical Translation
Authors: Shureiqi Imad , Baron John A.
Published: March 2011

Curcumin exhibits significant antitumorigenic activity in various preclinical models; data supporting its chemopreventive activity in humans, however, are lacking. To our knowledge, the first published results of a phase II chemoprevention study of curcumin are reported in this issue of the journal by Carroll and colleagues (beginning on page 354), who examined the effects of oral curcumin on various putative biomarkers of colonic tumorigenesis in smokers. This perspective discusses the potential significance and limitations of the current study findings in addressing the question of whether curcumin is clinically active as a chemopreventive agent. Cancer Prev Res; 4(3); 296–8. ©2011 AACR.
Circulating Leukocyte and Carotid Atherosclerotic Plaque Telomere Length
Authors: Huzen Jardi , Peeters Wouter , de Boer Rudolf A. , Moll Frans L. , Wong Liza S.M. , Codd Veryan , de Kleijn Dominique P.V. , de Smet Bart J.G.L. , van Veldhuisen Dirk J. , Samani Nilesh J. , van Gilst Wiek H. , Pasterkamp Gerard , van der Harst Pim
Published: March 2011

Objective—

Shorter leukocyte telomeres are associated with atherosclerosis and predict future heart disease. The goal of the present study was to determine whether leukocyte telomere length is related to atherosclerotic plaque telomere length and whether it is associated with plaque characteristics or recurrence of disease.

Methods and Results—

Telomere length was measured by real-time quantitative polymerase chain reaction in atherosclerotic plaques and leukocytes in patients with carotid atherosclerosis undergoing carotid endarterectomy (n=684) and of leukocytes in age- and gender-balanced subjects without clinical atherosclerosis (n=780). Leukocyte telomere length was shorter in patients versus controls (0.99 [interquartile range (IQR): 0.79 to 1.26] versus 1.06 [0.80 to 1.39];
P
=0.0007). Plaque telomeres were longer than leukocyte telomeres (1.42 [IQR: 1.21 to 1.77] versus 1.01 [IQR: 0.75 to 1.34];
P
<1.00×10
−6
) and independent of age. Leukocyte and plaque telomere length were only weakly correlated (correlation coefficient
r
2
=0.04,
P
=0.03). Patients, whose plaques showed marked macrophage infiltration and large lipid core, had longer plaque telomeres (1.61 [IQR: 1.32 to 2.04] versus 1.40 [IQR: 1.15 to 1.57];
P
=0.006) and shorter leukocyte telomeres (0.88 [IQR: 0.75 to 1.20] versus 1.03 [IQR: 0.83 to 1.34];
P
=0.02). Plaque telomere length was associated with restenosis 1 year after endarterectomy (OR 1.58±0.206;
P
=0.026 per SD decrease of plaque telomere length).

Conclusion—

Leukocyte telomere length is associated with the presence of atherosclerotic carotid plaques but is not a proxy for local plaque telomere length. Plaque telomere length is related to plaque characteristics and development of restenosis following endarterectomy.
Bone Marrow–Derived Smooth Muscle–Like Cells Are Infrequent in Advanced Primary Atherosclerotic Plaques but Promote Atherosclerosis
Authors: Yu Haixiang , Stoneman Victoria , Clarke Murray , Figg Nichola , Xin Hong-Bo , Kotlikoff Michael , Littlewood Trevor , Bennett Martin
Published: March 2011

Objective—

Although vascular smooth muscle cells (VSMCs) provide the major structural integrity of atherosclerotic plaques, their origin has been questioned. In particular, although some studies identified plaque VSMCs originating from bone marrow or peripheral blood, their frequency is controversial and their function unknown. We used genetic tracking of cell fate through smooth muscle cell (SMC)–specific LacZ reporter activity and VSMC-selective apoptosis to investigate the frequency, distribution, and role of marrow-derived VSMCs in atherogenesis.

Methods and Results—

Cultured mouse bone marrow–derived smooth muscle–like cells expressed SMC markers and functional SMC promoter-driven transgenes over time. Transplantation of apolipoprotein E (ApoE)
−/−
mice with smooth muscle myosin heavy chain–Cre/ROSA26R/ApoE
−/−
marrow showed that 0.7±0.14% cells expressed LacZ in atherosclerotic plaques, located superficially in early plaques, and in necrotic cores but not fibrous caps of advanced lesions. Cells expressing both progenitor and SMC markers showed a similar distribution and frequency. Apoptosis of marrow-derived SMC-like cells transplanted from SM22α–human diphtheria toxin receptor/ApoE
−/−
mice retarded atherogenesis, with reduced plaque macrophage content. Cultured marrow-derived SMC-like cells secreted proinflammatory cytokines and promoted macrophage migration, VSMC proliferation, and collagen synthesis.

Conclusion—

Bone marrow–derived SMC-like cells are infrequent in advanced primary atherosclerotic plaques and absent in fibrous caps. However, these cells secrete proinflammatory cytokines and mitogens and promote atherosclerosis.
Detection of Hypoxia by [
18
F]EF5 in Atherosclerotic Plaques in Mice
Authors: Silvola Johanna M.U. , Saraste Antti , Forsback Sarita , Laine V. Jukka O. , Saukko Pekka , Heinonen Suvi E. , Ylä-Herttuala Seppo , Roivainen Anne , Knuuti Juhani
Published: March 2011

Objective—

Atherosclerotic plaques with large lipid cores and inflammation contain regions of hypoxia. We examined the uptake of 2-(2-nitro-
1
H-imidazol-1-yl)-
N
-(2,2,3,3,3-pentafluoropropyl) acetamide ([
18
F]EF5), a specific marker of hypoxia labeled for positron emission tomography, in mouse atherosclerotic plaques.

Methods and Results—

Atherosclerotic mice of 2 different genetic backgrounds (low-density lipoprotein receptor
−/−
apolipoprotein B
100/100
and insulin-like growth factor II/low-density lipoprotein receptor
−/−
apolipoprotein B
100/100
) were first fed a Western diet to induce development of plaques with variable phenotypes and then injected with [
18
F]EF5. C57BL/6N mice served as controls. Aortas were dissected for biodistribution studies, autoradiography, histology, and immunohistochemistry. Uptake of [
18
F]EF5 was significantly higher in the aortas of mice with large atherosclerotic plaques than in the C57BL/6N controls. Furthermore, autoradiography demonstrated, on average, 2.0-fold higher [
18
F]EF5 uptake in atherosclerotic plaques than in the adjacent normal vessel wall. Hypoxia in plaques was verified by using an EF5 adduct-specific antibody and pimonidazole. The blood clearance of [
18
F]EF5 was slow, with blood radioactivity remaining relatively high up to 180 minutes after injection.

Conclusion—

Large atherosclerotic plaques in mice contained hypoxic areas and showed uptake of [
18
F]EF5. Despite its slow blood clearance, the high uptake of [
18
F]EF5 in plaques suggested that plaque hypoxia is a potential target for identifying high-risk plaques noninvasively.
Macrophage-Derived Tumor Necrosis Factor-α Is an Early Component of the Molecular Cascade Leading to Angiogenesis in Response to Aortic Injury
Authors: Ligresti Giovanni , Aplin Alfred C. , Zorzi Penelope , Morishita Ann , Nicosia Roberto F.
Published: March 2011

Objective—

The goal of this study was to define the role of tumor necrosis factor-α (TNFα) in the cascade of gene activation that regulates aortic angiogenesis in response to injury.

Methods and Results—

Angiogenesis was studied by culturing rat or mouse aortic rings in collagen gels. Gene expression was evaluated by quantitative reverse transcription–polymerase chain reaction, microarray analysis, immunocytochemistry, and ELISA. TNFα gene disruption and recombinant TNFα or blocking antibodies against vascular endothelial growth factor (VEGF) or TNF receptors were used to investigate TNFα-mediated angiogenic mechanisms. Resident aortic macrophages were depleted with liposomal clodronate. Angiogenesis was preceded by overexpression of TNFα and TNFα-inducible genes. Studies with isolated cells showed that macrophages were the main source of TNFα. Angiogenesis, VEGF production, and macrophage outgrowth were impaired by TNFα gene disruption and promoted by exogenous TNFα. Antibody-mediated inhibition of TNF receptor 1 significantly inhibited angiogenesis. The proangiogenic effect of TNFα was suppressed by blocking VEGF or by ablating aortic macrophages. Exogenous TNFα, however, maintained a limited proangiogenic capacity in the absence of macrophages and macrophage-mediated VEGF production.

Conclusion—

Overexpression of TNFα is required for optimal VEGF production and angiogenesis in response to injury. This TNFα/VEGF-mediated angiogenic pathway requires macrophages. The residual capacity of TNFα to stimulate angiogenesis in macrophage-depleted aortic cultures implies the existence of a VEGF-independent alternate pathway of TNFα-induced angiogenesis.
Propagation of the Sinus Impulse Into the Koch Triangle and Localization, Timing, and Origin of the Multicomponent Potentials Recorded in This Area
Authors: Pandozi Claudio , Ficili Sabina , Galeazzi Marco , Lavalle Carlo , Russo Maurizio , Pandozi Angela , Venditti Franco , Pristipino Christian , Verbo Brunella , Santini Massimo
Published: March 2011

Background—

The presence of a conduction block at the level of the Koch triangle (KT) and the origin of the multicomponent potentials inside this area are controversial issues. We investigated the propagation of the sinus impulse into the KT and the characteristics of multicomponent potentials recorded in that area in patients with and without atrioventricular nodal reentrant tachycardia (AVNRT).

Methods and Results—

Thirty-two patients (16 with AVNRT, 16 without AVNRT) underwent a sinus rhythm electroanatomic mapping of the right atrium (RA). Conduction velocities in the RA and in the KT were evaluated quantitatively on activation maps and qualitatively on isochronal and propagation maps. The presence, location, and timing of different types of multicomponent potentials were evaluated. A mean of 149±44 points were sampled in the RA, whereas a mean of 79±21 points were collected inside the KT. Propagation block at the level of crista terminalis was not found in any patient, whereas slow conduction inside the KT was found in all (median conduction velocity, 122 cm/s [110 to 135 cm/s] outside KT versus 60 cm/s [48 to 75 cm/s] inside KT;
P
<0.0001). Jackman potentials were identified inside KT in almost all the patients and were invariably found on the line of collision between the wavefronts activating the KT in opposite directions.

Conclusions—

No conduction block was detected inside the KT in patients with and without AVNRT. Conduction slowing was demonstrated during propagation of the sinus impulse inside the KT. The genesis of the Jackman potential may be related to the collision of the wavefronts activating KT in opposite directions.
FKBP12 Is a Critical Regulator of the Heart Rhythm and the Cardiac Voltage-Gated Sodium Current in Mice
Authors: Maruyama Mitsunori , Li Bai-Yan , Chen Hanying , Xu Xuehong , Song Long-Sheng , Guatimosim Silvia , Zhu Wuqiang , Yong Weidong , Zhang Wenjun , Bu Guixue , Lin Shien-Fong , Fishbein Michael C. , Lederer W. Jonathan , Schild John H. , Field Loren J. , Rubart Michael , Chen Peng-Sheng , Shou Weinian
Published: March 2011

Rationale:

FK506 binding protein (FKBP)12 is a known
cis-trans
peptidyl prolyl isomerase and highly expressed in the heart. Its role in regulating postnatal cardiac function remains largely unknown.

Methods and Results:

We generated FKBP12 overexpressing transgenic (αMyHC-FKBP12) mice and cardiomyocyte-restricted FKBP12 conditional knockout (FKBP12
f/f
/αMyHC-Cre) mice and analyzed their cardiac electrophysiology in vivo and in vitro. A high incidence (38%) of sudden death was found in αMyHC-FKBP12 mice. Surface and ambulatory ECGs documented cardiac conduction defects, which were further confirmed by electric measurements and optical mapping in Langendorff-perfused hearts. αMyHC-FKBP12 hearts had slower action potential upstrokes and longer action potential durations. Whole-cell patch-clamp analyses demonstrated an ≈80% reduction in peak density of the tetrodotoxin-resistant, voltage-gated sodium current
I
Na
in αMyHC-FKBP12 ventricular cardiomyocytes, a slower recovery of
I
Na
from inactivation, shifts of steady-state activation and inactivation curves of
I
Na
to more depolarized potentials, and augmentation of late
I
Na
, suggesting that the arrhythmogenic phenotype of αMyHC-FKBP12 mice is attributable to abnormal
I
Na
. Ventricular cardiomyocytes isolated from FKBP12
f/f
/αMyHC-Cre hearts showed faster action potential upstrokes and a more than 2-fold increase in peak
I
Na
density. Dialysis of exogenous recombinant FKBP12 protein into FKBP12-deficient cardiomyocytes promptly recapitulated alterations in
I
Na
seen in αMyHC-FKBP12 myocytes.

Conclusions:

FKBP12 is a critical regulator of
I
Na
and is important for cardiac arrhythmogenic physiology. FKPB12-mediated dysregulation of
I
Na
may underlie clinical arrhythmias associated with FK506 administration.
Cycling Induced by Electrical Stimulation Improves Motor Recovery in Postacute Hemiparetic Patients
Authors: Ambrosini Emilia , Ferrante Simona , Pedrocchi Alessandra , Ferrigno Giancarlo , Molteni Franco
Published: March 2011

Background and Purpose—

This study assessed whether cycling induced by functional electrical stimulation (FES) was more effective than passive cycling with placebo stimulation in promoting motor recovery and walking ability in postacute hemiparetic patients.

Methods—

In a double-blind, randomized, controlled trial, 35 patients were included and randomized to receive FES-induced cycling training or placebo FES cycling. The 4-week treatment consisted of 20 sessions lasting 25 minutes each. Primary outcome measures included the leg subscale of the Motricity Index and gait speed during a 50-meter walking test. Secondary outcomes were the Trunk Control Test, the Upright Motor Control Test, the mean work produced by the paretic leg, and the unbalance in mechanical work between paretic and nonparetic legs during voluntary pedaling. Participants were evaluated before training, after training, and at 3- to 5-month follow-up visits.

Results—

No significant differences were found between groups at baseline. Repeated-measures ANOVA (
P
<0.05) revealed significant increases in Motricity Index, Trunk Control Test, Upright Motor Control Test, gait speed, and mean work of the paretic leg after training and at follow-up assessments for FES-treated patients. No outcome measures demonstrated significant improvements after training in the placebo group. Both groups showed no significant differences between assessments after training and at follow-up. A main effect favoring FES-treated patients was demonstrated by repeated-measures ANCOVA for Motricity Index (
P
<0.001), Trunk Control Test (
P
=0.001), Upright Motor Control Test (
P
=0.005), and pedaling unbalance (
P
=0.038).

Conclusions—

The study demonstrated that 20 sessions of FES cycling training significantly improved lower extremity motor functions and accelerated the recovery of overground locomotion in postacute hemiparetic patients. Improvements were maintained at follow-up.
Safety and Tolerability of Early Noninvasive Ventilatory Correction Using Bilevel Positive Airway Pressure in Acute Ischemic Stroke
Authors: Tsivgoulis Georgios , Zhang Yi , Alexandrov Anne W. , Harrigan Mark R. , Sisson April , Zhao Limin , Brethour Mary , Cava Luis , Balucani Clotilde , Barlinn Kristian , Patterson Damon E. , Giannopoulos Sotirios , DeWolfe Jennifer , Alexandrov Andrei V.
Published: March 2011

Background and Purpose—

Hypercapnia can induce intracranial blood-flow steal from ischemic brain tissues, and early initiation of noninvasive ventilator correction (NIVC) may improve cerebral hemodynamics in acute ischemic stroke. We sought to determine safety and tolerability of NIVC initiated on hospital admission without polysomnography study.

Subjects and Methods—

Consecutive acute ischemic stroke patients were evaluated for the presence of a proximal arterial occlusion, daytime sleepiness, or history of obstructive sleep apnea, and acceptable pulse oximetry readings while awake (96%–100% on 2 to 4 L supplemental oxygen delivered by nasal cannula). NIVC was started on hospital admission as standard of care when considered necessary by treating physicians. NIVC was initiated using bilevel positive airway pressure at 10 cmH
2
O inspiratory positive airway pressure and 5 cmH
2
O expiratory positive airway pressure in combination with 40% fraction of inspired oxygen. All potential adverse events were prospectively documented.

Results—

Among 356 acute ischemic stroke patients (median NIHSS score, 5; interquartile range, 2–13), 64 cases (18%) received NIVC (median NIHSS score, 12; interquartile range, 6–17). Baseline stroke severity was higher and proximal arterial occlusions were more frequent in NIVC patients compared to the rest (
P
<0.001). NIVC was not tolerated by 4 patients (7%). Adverse events in NIVC included vomiting (n=1), aspiration pneumonia (n=1), respiratory failure/intubation (n=1), hypotension requiring pressors (n=1), and facial skin breakdown (n=3). The in-hospital mortality rate was 13% in NIVC patients and 8% in the rest (
P
=0.195). Neurological improvement during hospitalization tended to be greater in the NIVC group (median NIHSS score decrease, 2 points; interquartile range, 0–4) compared to the rest (median NIHSS score decrease, 1; interquartile range, 0–2;
P
=0.078).

Conclusions—

In acute ischemic stroke patients with proximal arterial occlusion and excessive sleepiness or obstructive sleep apnea, NIVC can be initiated early with good tolerability and a relatively small risk of serious complications.